micromachines

Review
Biomaterials Meet Microfluidics: From Synthesis
Technologies to Biological Applications
Jingyun Ma 1,2 , Yachen Wang 1,2 and Jing Liu 1,2, *
1 Regenerative Medicine Center, the First Affiliated Hospital of Dalian Medical University,
Dalian 116011, China; majingyun@dmu.edu.cn (J.M.); wangyachen100@163.com (Y.W.)
2 Stem Cell Clinical Research Center, the First Affiliated Hospital of Dalian Medical University,
Dalian 116011, China
* Correspondence: liujing@dmu.edu.cn; Tel.: +86-411-83635963-2170

Received: 7 July 2017; Accepted: 14 August 2017; Published: 19 August 2017

Abstract: Microfluidics is characterized by laminar flow at micro-scale dimension, high surface to

volume ratio, and markedly improved heat/mass transfer. In addition, together with advantages
of large-scale integration and flexible manipulation, microfluidic technology has been rapidly
developed as one of the most important platforms in the field of functional biomaterial synthesis.
Compared to biomaterials assisted by conventional strategies, functional biomaterials synthesized by
microfluidics are with superior properties and performances, due to their controllable morphology
and composition, which have shown great advantages and potential in the field of biomedicine,
biosensing, and tissue engineering. Take the significance of microfluidic engineered biomaterials
into consideration; this review highlights the microfluidic synthesis technologies and biomedical
applications of materials. We divide microfluidic based biomaterials into four kinds. According
to the material dimensionality, it includes: 0D (particulate materials), 1D (fibrous materials),
2D (sheet materials), and 3D (construct forms of materials). In particular, micro/nano-particles
and micro/nano-fibers are introduced respectively. This classification standard could include all of
the microfluidic biomaterials, and we envision introducing a comprehensive and overall evaluation
and presentation of microfluidic based biomaterials and their applications.

Keywords: functional biomaterials; microfluidics; controllable synthesis; biological applications

1. Introduction
Since the 1990s, no matter in the field of natural science, nor engineering technology,
miniaturization has become one of the general development trends [1–3]. A microfluidic system,
namely, lab-on-a-chip, is a multifunctional platform which integrates basic operating units involved
in the fields of chemistry and biology, such as sample preparation, reaction, separation, detection,
and cell culture, separation, lysis, into a chip, within an area of a few square centimeters [4,5]. In this
system, the micro-structured units and controllable fluidics constitute the network, which work as
conventional chemical or biological laboratories [6,7]. The idea of microfluidics fits well with the
concept of miniaturization, and thanks to its interdisciplinary advantages, it has been widely applied
in fields such as engineering, physics, chemistry, microscopy, and biotechnology [8–10].
Biomaterials refer to a class of materials that have specific biological functions.
Generally, biomaterials include any substance engineered to be a part of the biological system, or play
a template role which could be sacrificed, either for a therapeutic purpose, or a diagnostic one [11,12].
Biomaterials science is a subject of cross disciplines, including medicine, biology, and chemistry [13,14].
With the progress of biomaterials science, considerable research has focused on micro/nanometer-scale
materials with complex structures, because the microscopic architectures enable biomaterials great
optimized properties [15]. Conventional bulk synthesis usually adopts the certain physical or

Micromachines 2017, 8, 255; doi:10.3390/mi8080255 www.mdpi.com/journal/micromachines

Micromachines 2017, 8, 255 2 of 29

chemical method (e.g., mechanical stirring) [16,17]. These methods generally generate products
with monotonous morphology, and the dispersity of the biomaterials and production process are
difficult to control. Although there have been efforts to improve the homogeneity of products, such
as DeSimone’s print technique [18,19] and membrane emulsification [20], the lack of manipulation
flexibility still limits the improvement of materials synthesis. In particular, for biomaterials that are
specific, such as “intelligent biomaterials” or “functional biomaterials”, the conventional approaches
are insufficient to meet their synthetic requirements. Therefore, it is extremely urgent to develop novel
synthesis technologies for functional biomaterials. At this point, characterized by the microscale and
rapid process, microfluidics, comes into sight and arouses the great interest of researchers.
Micro/nanometer-scale functional biomaterials synthesized from the microfluidic platform are
of different configurations, complex structures, and novel properties in flexible and easily prepared
way [21,22]. Compared to conventional synthetic methods, the advantages of microfluidic synthesis
lie in following aspects. The biomaterial size, morphology, and composition are easily controlled,
resulting in superior properties. In the micro-scale system, the reaction could be accelerated due to
low consumption of reagents, rapid heat transfer, and mass transfer. The internal reaction conditions
are stable, with no cross contamination. The preparation is of spatiotemporal resolution, and with
controllable addition of the reagent, which is beneficial to the process of multi-step and multi-reagent
synthesis. Through the scale integration of a microfluidic system, flexible manipulation, and equip
automation, the complex reaction process can be simplified. Due to the minute volume of the
microfluidic chips, and match of most chips’ materials to the microscopic observations, real-time
monitoring of the reaction process in the chip could be realized, which helps to clarify the synthesis
mechanism [23,24]. Therefore, the use of microfluidic technology to design and prepare functional
biomaterials has become a hot topic recently, moreover, will continue to bring infinite possibilities for
the future development of the areas of materials science and biology [25,26].
To date, there have been several excellent reviews on the microfluidic fabrication of materials, from
different points of view [27–29], which mainly deal with certain microfluidic technologies (e.g., droplet
microfluidic) or specific forms of materials (e.g., Janus particles). A comprehensive review pinpointing
the microfluidic synthesis and bio-applications of materials is still lacking. In this review, we summarize
the microfluidic methods for preparation and applications of all kinds of biomaterials, both at
micro-scale and nano-scale, and in definite classification, from 0D to 3D forms. Herein, classical and
recent achievements in the biomaterials engineered from microfluidics are presented and categorized.
According to the material dimension, namely, dot, line, face, and body, microfluidic biomaterials
are classified into four main categories, that include zero-dimensional particulate materials (0D),
one-dimensional fibrous materials (1D), two-dimensional sheet materials (2D), and three-dimensional
construct forms of materials (3D), as shown in Figure 1. In particular, micro/nano-particles
and micro/nano-fibers are introduced respectively. More specifically, 0D materials refer to that
with sizes in every direction at the same order of magnitude, presenting “particle” appearance.
For micro/nano-particles, the sizes in every direction are all at micro/nano scale, respectively. For 1D
material, size in one direction is several orders greater than those in other directions, presenting “fiber”
appearance. For micro/nano-fibers, the obvious size in certain direction maybe at millimeter even
meter-scale, while the sizes of other directions are micro/nano scale, respectively. For 2D material,
size in one direction is several orders less than those in other directions, presenting “sheet” appearance.
Similar to 0D materials, 3D materials refer to that with sizes in every direction at almost the same
order of magnitude, presenting “construct” appearance. However, the sizes of 3D materials in at least
two directions are at millimeter scale. Compared to 2D material, 3D materials are more stereoscopic.
 Micromachines
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2017, 255255 3 of 2929
3 of

Core-shell
structured
particle
Special-shaped Porous
particle particle
Spherical Composite
particle 0D: Particulate biomaterials particle

Microfluidic
synthesis

1D: Fibrous biomaterials 3D: Construct forms

of biomaterials

2D: Sheet biomaterials

Figure
Figure 1. 1. Schematic
Schematic diagram
diagram of of
thethe classification
classification of of biomaterials
biomaterials engineered
engineered from
from microfluidics.
microfluidics.

2.2.
Particulate Biomaterials
Particulate Synthesis
Biomaterials and
Synthesis Applications
and Applications

Traditional
Traditionalmethods
methodsofofpreparing
preparingparticulate
particulatematerials
materialsincludeincludemechanical
mechanicalagitation,
agitation,emulsion
emulsion
polymerization,
polymerization, seeding
seeding polymerization,
polymerization, precipitation,
precipitation, etc.,etc.,
however,
however,due todue thetobroad particle
the broad size
particle
distribution and uncontrollable morphology, their applications are greatly restricted. Because of the
size distribution and uncontrollable morphology, their applications are greatly restricted. Because
advantages of uniform
of the advantages of particle
uniformsize and controllable
particle morphology,
size and controllable particulate particulate
morphology, biomaterialsbiomaterials
synthesis
from microfluidics has become a hot topic in recent years [30,31]. In general, microfluidic
synthesis from microfluidics has become a hot topic in recent years [30,31]. In general, microfluidic
technologies
technologies forforparticulate
particulate biomaterials
biomaterials synthesis
synthesis areare composed
composed ofofdroplet
dropletmicrofluidics
microfluidics [32]
[32]and
and
photolithography
photolithography[33]. [33].
Droplet
Dropletmicrofluidics
microfluidicsuses usesfluidic
fluidicmanipulation
manipulation atatthethemicro
microscale,
scale,dispersing
dispersingone oneliquid
liquidintointo
another
anotherliquid
liquidthat
thatisisimmiscible,
immiscible, generating independent
independentliquid liquidunits
unitsininthe
the microfluidic
microfluidic channels
channels [34].
[34]. Correspondingly,
Correspondingly, a multi-spatial
a multi-spatial reaction
reaction synthesis
synthesis systemsystem is constructed,
is constructed, which iswhich
located is on
located on
the inside
the
of inside of the
the liquid drop,liquid drop, the
the liquid drop liquid
edge,drop edge,
and the and drop
liquid the liquid
intervaldrop
[35].interval
In droplet[35].microfluidics
In droplet
microfluidics
synthesis, the synthesis,
dimensions,the dimensions,
shape, and shape, and monodispersity
monodispersity of the can
of the droplets, droplets, can be precisely
be precisely controlled,
controlled, and enable the method one of the most commonly used microfluidic material synthesis
and enable the method one of the most commonly used microfluidic material synthesis means.
means. The principles
The principles and chip anddesigns
chip designs for droplet
for droplet generationgeneration are asinshown
are as shown Figurein2,Figure
including2, including
T-junction
T-junction
(Figure 2a), (Figure 2a), flow-focusing
flow-focusing (Figure 2b),(Figure 2b), and
and coaxial (Figure coaxial (Figure 2c)
2c) structured chip.structured
In T-junctionchip. chip,
In
T-junction chip, the dispersed phase flows from a vertical channel to a horizontal channel, filled with
the dispersed phase flows from a vertical channel to a horizontal channel, filled with the continuous
the continuous
phase. Underphase. Under theaction
the combined combined action
of shear of shear
force force andpressure
and extrusion extrusionboth pressure
from both from
continuous
continuous phase, monodispersed
phase, monodispersed droplets are droplets are generated.
generated. In the flow-focusing
In the flow-focusing chip, the phase
chip, the dispersed dispersedflows
phase flows from the middle channel, and suffers extrusion force of the continuous phase from both
from the middle channel, and suffers extrusion force of the continuous phase from both sides.
sides. The dispersed
The dispersed phase phase experiences
experiences stretching
stretching and breakage,
and breakage, leadingleading to droplet
to droplet formation.
formation. WhileWhile
for the
forcoaxial
the coaxial structured
structured chip,
chip, the the dispersed
dispersed phase phase
channel channel is embedded
is embedded in the continuous
in the continuous phase
phase channel,
channel,
and theand the dispersed
dispersed phaseparallel
phase flows flows parallel to the continuous
to the continuous phase towards
phase towards the same thedirection.
same direction.
As well,
Asthewell, the dispersed
dispersed phase is phase
brokenisinto
broken into In
droplets. droplets. In the microchannels,
the microchannels, droplet isgeneration
droplet generation influencedisby
influenced by the microchannel
the microchannel construction,
construction, viscosity viscosity
and flow velocityandofflow velocity
the two phases,of the
andtwo phases,tension
interfacial and
interfacial
between tension between
each adjacent each adjacent
phase. Therefore, phase. Therefore, and
the dimension the dimension
productionand rateproduction
of the dropletsrate ofcanthebe
droplets canby
controlled be adjusting
controlledthe byabove
adjusting the above
parameters. In parameters.
addition, double In addition,
or evendouble
multiple oremulsions
even multiple could
emulsions could be generated for particulate material synthesis with core-shell or multi-core
structure, through the composite design of microchannels.
Micromachines 2017, 8, 255 4 of 29

be generated for particulate material synthesis with core-shell or multi-core structure, through the
Micromachines 2017, 8, 255 4 of 29
composite design of microchannels.

a b c d
Continuous Continuous phase Monomer
Dispersed
phase phase
Monomer
Microscope
Objective
Continuous phase Dispersed Mask
phase Dispersed phase UV

Figure 2.
Figure 2. Microfluidic
Microfluidic technologies
technologies for
for the
the synthesis
synthesis of
of particulate
particulate materials.
materials. The
The principles
principles and
and chip
chip
designs with different flow regimes for droplet generation, including T-junction (a); flow-focusing
designs with different flow regimes for droplet generation, including T-junction (a); flow-focusing (b);
(b); and
and coaxial
coaxial (c) structured
(c) structured chip;chip; (d) Photolithography
(d) Photolithography applied
applied in microfluidic
in microfluidic synthesis.
synthesis.

Particles from
Particles fromdroplet
droplet microfluidics
microfluidics are mainly
are mainly limitedlimited to spherical
to spherical or simpleor simple variations,
spherical spherical
variations,
such such ascylindrical
as hemisphere, hemisphere, cylindrical
or pie-like. Besides,or inpie-like.
some cases, Besides,
a chemicalin some cases, is
modification a needed
chemical to
modification is needed to guarantee a continuous and smooth formation
guarantee a continuous and smooth formation and flow of the droplets in the microchannel. In order and flow of the droplets in
theimprove
to microchannel. In orderoftodroplet
the limitations improve the limitations
microfluidics, of droplet microfluidics,
photolithography is put forward photolithography
to microfluidic is
put forward
synthesis, whichto microfluidic synthesis, which
combines microfluidics and combines microfluidics
photolithography [36], asand photolithography
shown in Figure 2d. [36], as
In this
shown inmonomer
method, Figure 2d. In this
solution thatmethod, monomer
is sensitive solution
to ultraviolet that
light is sensitiveinto
is introduced to ultraviolet light is
the single channel,
introduced
and one side into
of the
the single
chip ischannel,
coveredand onemask
by the side of thecontains
that chip is covered
the design by the mask After
pattern. that contains
ultraviolet the
design pattern. After ultraviolet irradiation, partial polymerization of
irradiation, partial polymerization of the monomer solution endows the materials with specific shapes. the monomer solution endows
the materials
According to thewith specific polymerization
monomer shapes. According tothe
rates, thefluid
monomer
can be polymerization
applied with lights rates, the fluid can
in continuous be
flow
applied
or with lightsmanner.
flow–stop–flow in continuous
Althoughflow or flow–stop–flow
photolithography enrichesmanner. Although
the material shape photolithography
of microfluidic
enriches the
synthesis, it ismaterial shape for
only suitable of microfluidic
light-sensitive synthesis,
materials. it is only suitable for light-sensitive materials.
The specific
The specific and and primary
primary shapeshape could
could be be given
given to to the
the biomaterials
biomaterials by by microfluidic
microfluidic technology,
technology,
however, regardless
however, regardless of of the
the particulate,
particulate, fibrous,
fibrous, sheet,
sheet, or or construct
construct forms forms of of materials,
materials, specific
specific
solidification methods are needed to get the final form of the material.
solidification methods are needed to get the final form of the material. Principles and methods of Principles and methods of
materials solidification
materials solidification involved
involved in microfluidic
in microfluidic synthesissynthesis include polymerization,
include polymerization, solvent
solvent volatilization
volatilization and solvent extraction, ionic crosslinking and chemical
and solvent extraction, ionic crosslinking and chemical crosslinking for organic materials, and inorganic crosslinking for organic
materials,reaction
chemical and inorganic chemical reaction
and self-assembly and self-assembly
for inorganic materials. for inorganic materials.
The polymerization
The polymerization reaction reaction is is the
the method
method for for preparing
preparing polymer
polymer functional
functional materials
materials using using
monomer as the raw material. The monomer solution in the microchannel
monomer as the raw material. The monomer solution in the microchannel is as the dispersed phase is as the dispersed phase
or continuous
or continuous phase, phase, and and solidified
solidified by by polymerization
polymerization into into aa particular
particular shape.
shape. According
According to to the
the
reaction triggering
reaction triggering modes, modes, the the polymerization
polymerization method method can can be divided into
be divided into light
light polymerization,
polymerization,
thermal polymerization
thermal polymerization and and polymerization
polymerization for for which
which only
only the the initiator
initiator is is needed,
needed, without
without external
external
light or
light or heat
heat stimulation.
stimulation.InInlight lightpolymerization,
polymerization, thethemonomer
monomer solution
solutionis exposed
is exposed to theto light and
the light
polymerized in the channel, and the materials are collected from the
and polymerized in the channel, and the materials are collected from the channel after basic curing. channel after basic curing. Heat
polymerization
Heat polymerization is triggered
is triggeredby heating
by heating thethepolymer
polymer system,
system, ororbybysimply
simplyreducing
reducingthe the temperature
temperature
of the
of the solution
solution with with high
high melting
melting point.
point. TheThe initiator
initiator polymerization
polymerization needs needs thethe addition
addition of of aa certain
certain
chemical that triggers the polymerization process. Crosslinking reactions,
chemical that triggers the polymerization process. Crosslinking reactions, such as ionic crosslinking such as ionic crosslinking
and chemical
and chemicalcrosslinking,
crosslinking, areare formed
formed between
between the polymer
the polymer chains chains and crosslinkers
and crosslinkers through
through covalent
covalent
or or noncovalent
noncovalent bonds. evaporation
bonds. Solvent Solvent evaporationand solventand solvent
extraction extraction both the
both utilize utilize the polymer
polymer as the
as the raw material. In the solvent evaporation method, the volatile
raw material. In the solvent evaporation method, the volatile solvent evaporates from the polymer solvent evaporates from the
polymer and
solution, solution, and theispolymer
the polymer solidified,is solidified, while for
while for solvent solvent extraction,
extraction, the solventthe solvent isbyextracted
is extracted another
by another chemical, and the polymer is separated out and solidified.
chemical, and the polymer is separated out and solidified. For microfluidic inorganic materials For microfluidic inorganic
materials synthesis,
synthesis, the raw materials
the raw materials experience experience various inorganic
various inorganic reactionsreactions in the microchannels,
in the microchannels, such as
such as hydrolysis,
hydrolysis, reduction, reduction, precipitation,
precipitation, etc. In particular,
etc. In particular, based on based on thisprinciple,
this reaction reaction aprinciple,
variety ofa
variety of nano-functional
nano-functional biomaterials could biomaterials
be synthesizedcould inbe thesynthesized
microfluidic device. in the Self-assembly
microfluidic refers device.to
Self-assembly refers to the particles or molecules being self-assembled, forming materials with more
complicated construction and multidimensional performance.
Accompanied by the development of technologies and theories as mentioned above,
microfluidics-based particulate biomaterials synthesis and applications have experienced a process,
from simple to complex. A series of particulate biomaterials, such as spherical particles, special
Micromachines 2017, 8, 255 5 of 29

the particles or molecules being self-assembled, forming materials with more complicated construction
and multidimensional performance.
Accompanied by the development of technologies and theories as mentioned above,
microfluidics-based particulate biomaterials synthesis and applications have experienced a process,
from simple to complex. A series of particulate biomaterials, such as spherical particles, special
shape particles, porous particles, core-shell structured particles, and even multi-component composite
particles, have been developed.

2.1. Particulate Biomaterials at Micro-Scale

2.1.1. Spherical Microparticles

In the absence of any external force, droplets tend to be spherical, to keep the minimum surface
energy and most stable condition, therefore, the synthesis of micro-spherical materials had been first
realized in the microfluidic system. In 2000, Nakajima and co-workers reported the first research
work on microsphere synthesis in the microfluidic device [37]. Based on the thermal polymerization
principle, firstly, at high temperatures, oil-in-water (O/W) droplets were generated. After the
processing of curing and freeze-drying, the lipid microspheres were with an average dimension of 20
µm, and variable coefficient less than 5%, which improved the particle size distribution effectively,
compared to conventional suspension polymerization suitable for microsphere synthesis above 10 µm.
Kumacheva and co-workers dispersed the polymer monomer into droplets on the microfluidic chip.
The droplets were turned into microspheres by ultraviolet light or heat-induced polymerization, as
shown in Figure 3a [38]; the droplets could be generated steadily, received the external stimulus in
turn, and transformed into polymer microspheres with a uniform particle size in a single dispersed
form. This microsphere synthesis is stable and simple, but only suitable for photosensitive or
heat-sensitive raw materials. Shibata and co-workers introduced a fluorescent dye with glucose
responsiveness into the monomer, and this functional monomer was emulsified and polymerized
in the chip to form gel microspheres [39]. The fluorescence intensity of microspheres injected into
the mice represented different levels of glucose in the body, and the real-time monitoring of glucose
concentration was achieved.
The preparation of biocompatible materials, which can be used for the drug release, cell cultivation,
and protein purification, has become one of the research hotspots. Among them, biocompatible calcium
alginate and chitosan materials are usually prepared based on crosslinking, namely, the polymer
solution is dispersed as an emulsion in the microchannel, and the crosslinking agent is then introduced,
and triggers the crosslinking reaction that allows the emulsions to solidify. Takeuchi and co-workers
prepared uniform calcium alginate microbeads according to the principle of internal gelation, which
were used for leukemia cell encapsulation [40]. As shown in Figure 3b, in the “T” structure of chip,
the sodium alginate was dispersed into droplets with cells and CaCO3 nanoparticles. By adding
continuous phase containing acetic acid in the downstream channel, CaCO3 nanoparticles inside the
droplets released the Ca2+ , and triggered the gelation reaction to form the microcapsules.
Microspheres prepared by the O/W liquid drop synthesis system have extensive applications
in the field of drug release. In general, biocompatible and biodegradable polymer materials,
such as poly lactic-co-glycolic acid (PLGA), polycaprolactone (PCL) and polylactic acid (PLA),
are dissolved in an organic reagent as the dispersed phase. After emulsion and solvent evaporation,
the microspheres are generated [41–43]. The microsphere particle size is determined by the size of
the droplets, and the concentration of the polymer. Take PLGA microspheres, for example, as shown
in Figure 3c. Firstly, PLGAwas dissolved in dimethylcarbonate, as the dispersed phase. At the
flow-focusing structure of chip, this fluid was emulsified into droplets by the continuous phase
containing 1% polyvinyl alcohol (PVA) as the surfactant. After complete volatilization of the solvent
dimethylcarbonate, PLGA microspheres were solidified.
Micromachines 2017, 8, 255 6 of 29

Utilizing the hydrolysis reaction of the inorganic oxide precursor, sol solution was generated
and emulsified into droplets, and after solvent dissipating, inorganic salt oxide microspheres were
synthesized; a typical example is the synthesis of SiO2 microspheres [44], as shown in Figure 3d. It is
Micromachines
expected that 2017, 8, 255
these 6 of 29
ordered mesoporous silica microspheres might be useful for biomolecule delivery.

50 μm

50 μm

d Emulsifier
Solvent diffusion

Surfactant

Precursor

3 μm 3 μm

Figure 3. Spherical microparticles prepared by droplet-based microfluidics. (a) Microspheres from

Figure 3. Spherical microparticles prepared by droplet-based microfluidics. (a) Microspheres from
ultraviolet
ultraviolet (UV)
(UV) or heat-induced polymerization
or heat-induced polymerization in microfluidic droplets;
in microfluidic droplets; (b)
(b) Calcium
Calcium alginate
alginate
microbeads
microbeads for cell encapsulation;
for cell encapsulation; (c)
(c) Oil-in-water
Oil-in-water (O/W)
(O/W) droplet
droplet synthesis
synthesis of
of poly
poly lactic-co-glycolic
lactic-co-glycolic
acid (PLGA)microspheres;
acid (PLGA) microspheres; (d) SiO
(d) SiO 2 microspheres generated based on sol-gel principle and
2 microspheres generated based on sol-gel principle and microfluidic
microfluidic droplets. Reproduced with
droplets. Reproduced with permission from permission from [38,40,41,44].
[38,40,41,44].

2.1.2. Special-Shaped Microparticles

2.1.2. Special-Shaped Microparticles
The acquisition of the spherical shape of the solid microspheres only relies on the most steady
The acquisition of the spherical shape of the solid microspheres only relies on the most steady
state of the droplets, and the synthesis is relatively simple and reflects research in the early
state of the droplets, and the synthesis is relatively simple and reflects research in the early exploration
exploration stage of microfluidic material synthesis. Accompanied by the development of
stage of microfluidic material synthesis. Accompanied by the development of microfluidic technology
microfluidic technology and growing demand of advanced biomaterials, researchers shifted their
and growing demand of advanced biomaterials, researchers shifted their gaze to the preparation
gaze to the preparation of special-shaped microparticles, such as the hemisphere, round cake,
crescent, polygon, and stick. Besides uniform material size, versatile shape and morphology of
materials are the more prominent advantages of microfluidic synthesis.
By the combination of the droplet microfluidic technology and microchannel configuration
design, deformation of the droplet was realized. After in situ light polymerization or refrigeration,
Micromachines 2017, 8, 255 7 of 29

of special-shaped microparticles, such as the hemisphere, round cake, crescent, polygon, and stick.
Besides uniform material size, versatile shape and morphology of materials are the more prominent
advantages of microfluidic synthesis.
By the combination of the droplet microfluidic technology and microchannel configuration
design, deformation of the droplet was realized. After in situ light polymerization or refrigeration,
Micromachines
non-spherical2017, 8, 255particles were achieved [45]. As shown in Figure 4a, when the droplets diameter
solid 7 of 29
was smaller or greater than both of the microchannel depth and width, the particles presented as
spherical
spherical or
or short
short bar;
bar; when
when thethe droplets
droplets diameter
diameter waswas greater
greater than
than the
the microchannel
microchannel depthdepth andand
smaller
smaller than
than the
the microchannel
microchannel width,
width, the
the particles
particles presented
presented asas aa round
round pie.
pie. The
The method
method illustrated
illustrated
visually thatbyby
visually that adjustment
adjustment ofdroplet
of the the droplet
size andsize and the microchannel
the microchannel size, special-shaped
size, special-shaped microparticles
microparticles could be formed by the squeezing effect. Taking advantage
could be formed by the squeezing effect. Taking advantage of local surface modification of local surface
of the chip
modification of the chip channel and two droplet forming units, Zhang and
channel and two droplet forming units, Zhang and co-workers obtained double emulsion droplets co-workers obtained
double emulsion droplets
oil-in-water-in-oil (O/W/O). oil-in-water-in-oil
By changing the (O/W/O). By changing
flow rates, the number the of
flow rates, oil
internal the cores
number couldof
internal oil cores
be adjusted. Under could be adjusted.
the synergistic Under
effect the synergistic
of geometric effect
limitations of geometric limitations
of microchannels and inhibited of
microchannels and inhibited interfacial polymerization, and by removing the
interfacial polymerization, and by removing the internal oil cores, meniscus or multi-foot hydrogelinternal oil cores,
meniscus or multi-foot
microparticles hydrogel microparticles
were synthesized [46]. were synthesized [46].

120 μm 30 μm 400 μm

Figure
Figure 4.
4. Special-shaped
Special-shapedmicroparticles
microparticlesprepared
prepared byby droplet-based
droplet-based microfluidics.
microfluidics. (a)
(a) Poly
Poly
tripropyleneglycol
tripropyleneglycol diacrylate
diacrylate particles
particles with
with different
different shapes
shapes determined
determined by
by the
the microchannel
microchannel
dimension;
dimension; (b)
(b) Polylactic
Polylactic acid
acid (PLA)
(PLA) crescent-shaped
crescent-shaped composite
composite microparticles
microparticles from
from core-shell
core-shell
microcapsules
microcapsulesininthe
themicrofluidic
microfluidicchip;
chip;(c)
(c) One-step
One-stepprepared
prepared doughnut-shaped
doughnut-shapedSiOSiO22microparticles.
microparticles.
Reproduced
Reproduced with
with permission
permission from
from [45,47,48].
[45,47,48].

Vladisavljevic
Vladisavljevicetetal.
al.developed
developedcore-shell microcapsules
core-shell microcapsulesin in
thethe
microfluidic chip,
microfluidic which
chip, could
which be
could
turned into crescent-shaped composite microparticles after drying, due to the collapse of
be turned into crescent-shaped composite microparticles after drying, due to the collapse of PLA PLA shells
encompassing water-rich
shells encompassing crescent
water-rich regions,
crescent as shown
regions, in Figure
as shown in Figure4b4b[47].
[47].These
These crescent-shaped
crescent-shaped
microwells could be used for cell trapping/immobilisation. Doughnut shaped SiO2 microparticles
were prepared in one-step by Douliez and co-workers. The reduction of solvent in silica gel caused a
change in gelation, and the curvature instability at the droplet interface induced depression in the
cross-flow direction, leading to the formation of a ring structure, as shown in Figure 4c [48].
Doyle and Kwon’s groups performed a series of works on the use of photolithography to
prepare microparticles. As shown in Figure 5, a monomer solution that was sensitive to ultraviolet
Micromachines 2017, 8, 255 8 of 29

microwells could be used for cell trapping/immobilisation. Doughnut shaped SiO2 microparticles
were prepared in one-step by Douliez and co-workers. The reduction of solvent in silica gel caused
a change in gelation, and the curvature instability at the droplet interface induced depression in the
cross-flow direction, leading to the formation of a ring structure, as shown in Figure 4c [48].
Doyle and Kwon’s groups performed a series of works on the use of photolithography to prepare
microparticles. As shown in Figure 5, a monomer solution that was sensitive to ultraviolet (UV) light
was introduced into the channel, and one side of the chip was covered with the mask that contains the
Micromachines
design pattern. 8, 255 the UV exposure, the monomer solution was partially polymerized, due to
2017,After 8 ofthe
29
patterned masking effect. Using flow photolithography, special-shaped microparticles in triangles,
microparticles
circles, squares,inpolygons,
triangles,and
circles, squares,
ring shapes polygons,
could and ring
be obtained shapes
in the could be
microfluidic chipobtained in the
[49]. Similarly,
microfluidic chip [49]. Similarly, gear shaped polyacrylamide-silicon composite particles
gear shaped polyacrylamide-silicon composite particles were generated [50]. Multilayer structured were
generated [50]. Multilayer structured microparticles could be fabricated by the combination
microparticles could be fabricated by the combination of photolithography and 3D design of the of
photolithography
chip [51]. and 3D design of the chip [51].

20 μm 20 μm 20 μm
50 μm 5 μm

20 μm 20 μm 20 μm

50 mm
20 μm 20 μm 20 μm

Figure 5. Special-shaped
Figure 5. Special-shaped microparticles
microparticles prepared
prepared by
by photolithography
photolithography -based
-based microfluidics.
microfluidics.
Reproduced with permission from [49–51].

2.1.3.
2.1.3. Core-Shell
Core-Shell Structural Microparticles
Structural Microparticles
Microparticles
Microparticles withwith micro-hierarchical
micro-hierarchical structure,
structure, suchsuch asas core-shell
core-shell andand porous structure, are
porous structure, are
highly
highly regarded
regardeddue dueto to
their advanced
their advanced properties. Firstly,Firstly,
properties. the successful syntheses
the successful of these materials
syntheses of these
with special
materials withconfigurations reflect the improvement
special configurations of modern synthetic
reflect the improvement of modern techniques.
synthetic Next, these
techniques.
materials
Next, thesefacilitate
materialsfurther understanding
facilitate of the mechanism
further understanding of microstructure
of the mechanism of microstructureformation, thus
formation,
encouraging
thus encouraginginspiration into into
inspiration the the
design of more
design of more innovative
innovative materials.
materials.Once
Oncemore,more,relative
relative to
to the
the
microparticles with simple structure, hierarchical structure endows materials
microparticles with simple structure, hierarchical structure endows materials with novel construction with novel
construction and superioroptical,
and superior mechanical, mechanical,
and otheroptical, and other
properties. properties.
Finally, Finally,
the excellent the excellent
material performancematerial
will
performance
greatly enrichwill
andgreatly
develop enrich and application
practical develop practical
in the application
biology field. in the biology field.
Core-shell
Core-shell microparticles
microparticles referrefer to
to particles
particles ofof which
which shell
shell and
and kernel
kernel areare made
made up up of different
of different
substances.
substances. TheTheshells
shellsare aregenerally
generally solid, and
solid, andthethe
kernel would
kernel be one
would be oneor more
or more of solid, liquid,
of solid, and
liquid,
gaseous states.
and gaseous The whole
states. The wholematerials consists
materials of special
consists structures
of special like yolk–egg
structures like yolk–eggwhite, yolk–shell
white, and
yolk–shell
empty
and emptyshell.shell.
The core–shell
The core–shell microparticles realizerealize
microparticles the internal hierarchy
the internal of theofmaterial.
hierarchy The
the material.
compartments
The compartments couldcould
be with be independent
with independentcarrying capacity
carrying and function,
capacity and at the
and function, andsame time,
at the the
same
compartments could participate in the overall function exertion of the
time, the compartments could participate in the overall function exertion of the material in thematerial in the synergetic
manner.
synergetic manner.
Synthesis
Synthesis ofofthethe core–shell
core–shell microparticles
microparticles commonly
commonly uses multi-emulsion
uses multi-emulsion dropletsdroplets as the
as the template.
template.
Figure 6a illustrates the schematic diagram of double emulsion droplet generation in theinchip.
Figure 6a illustrates the schematic diagram of double emulsion droplet generation the
chip. O/W/O
Take Take O/W/O double
double emulsion
emulsion droplets
droplets forfor example,
example, in in ordertotoform
order formoil oilinin water
water droplets
droplets in in
the first emulsion
the first emulsion unit,
unit, hydrophilic
hydrophilic modification
modification should
should be be performed
performed for for the
the first
first emulsion
emulsion unit,
unit,
whilst hydrophobic
whilst hydrophobic modification
modification shouldshould bebe performed
performed for for the second emulsion
the second emulsion unit,unit, to enable the
to enable the
intermediate waterphase
intermediate water phase change
change fromfrom the continuous
the continuous phase phase to the dispersed
to the dispersed phase, forphase, for the
the generation
generation and stable existence of the double emulsion [52]. As shown in Figure 6b, in a capillary
microfluidic device with nested structures, the double emulsion could be one-step generation.
Similarly, by adding the emulsion unit in the chip, the synthesis of multiple emulsion droplets can
be realized.
The O/W/O double emulsion droplets with hydrogel precursor shell and magnetic monomer
Micromachines 2017, 8, 255 9 of 29

and stable existence of the double emulsion [52]. As shown in Figure 6b, in a capillary microfluidic
device with nested structures, the double emulsion could be one-step generation. Similarly, by adding
the emulsion unit in the chip, the synthesis of multiple emulsion droplets can be realized.
The O/W/O double emulsion droplets with hydrogel precursor shell and magnetic monomer core
were used for microparticle synthesis [53]. As shown in Figure 6c, the shell part was photopolymerized,
and the core was thermal polymerized. Under the external magnetic field, the microparticles
could be manipulated via the magnetic core. Zhao et al. dispersed quantum dots (QDs) and
magnetic nanoparticles into photochemical resin, respectively, and the two independent oil cores
were wrapped
Micromachines 2017,in the same hydrogel shell [54], as shown in Figure 6d. These microparticles 9were
8, 255 of 29
with functions of transmitting optical information by the QDs cores, magnetic response characteristics
characteristics
by the magnetic bynanoparticle
the magnetic nanoparticle
cores, cores, and biocompatibility
and biocompatibility by the hydrogelby the hydrogel
shells. shells. to
It was effective It
was
avoideffective
contacttowithavoid contact
toxic with toxic
materials in thematerials
internal in the internal
matrix. matrix.
Anderson Anderson
et al. developedet al.microfluidic
developed
microfluidic
microcapsulesmicrocapsules incorporating
incorporating colloidal colloidal
nanosensors, whichnanosensors,
could be usedwhich could besensing
for biomolecular used [55].
for
biomolecular
The microcapsulessensingwith
[55].polyethylene
The microcapsules
glycol with
shell polyethylene glycolcan
and liquid cores shell and liquid
selectively cores can
encapsulate
selectively
nanosensors encapsulate
within thenanosensors within thefree
core, and guarantee core, and guarantee
diffusion free diffusion
of molecules. Glucoseofand molecules.
heparin
Glucose and heparin
were detected were detected quantum
by glucose-responsive by glucose-responsive quantum
dots/gold nanorods, anddots/gold nanorods, gold
heparin-responsive and
heparin-responsive
nanorods, respectively. gold nanorods, respectively.

a b

c d

50 μm 200 μm

Figure 6. Core-shell
Figure 6. Core-shell structure
structure microparticles
microparticles prepared
prepared by
by multiple
multiple emulsion-based
emulsion-based microfluidics.
microfluidics.
(a,b) Designs of microfluidic devices for the preparation of double emulsion
(a,b) Designs of microfluidic devices for the preparation of double emulsion droplets; droplets; (c)
(c) Microparticle
Microparticle synthesis with magnetic core and hydrogel shell from double-emulsion
synthesis with magnetic core and hydrogel shell from double-emulsion microfluidic microfluidic
droplets;
droplets; (d) Microparticles
(d) Microparticles with two independent
with two independent oil cores
oil cores wrapped in thewrapped
hydrogel in theReproduced
shell. hydrogel shell.
with
Reproduced with[53,54].
permission from permission from [53,54].

Core-shell structure microparticles could also be generated based on phase separation. W/O
Core-shell structure microparticles could also be generated based on phase separation.
droplets were formed in the chip, and the separation reagents in the continuous phase spread into
W/O droplets were formed in the chip, and the separation reagents in the continuous phase spread into
the polyethylene glycol diacrylate (PEGDA) water phase, forming the core-shell structure. The phase
the polyethylene glycol diacrylate (PEGDA) water phase, forming the core-shell structure. The phase
separation process was shown in Figure 7a [56]. By adjusting the concentration of PEGDA,
separation process was shown in Figure 7a [56]. By adjusting the concentration of PEGDA, multi-wall
multi-wall microspheres could be obtained, which was known as the “onion” structure. Zhang et al.
microspheres could be obtained, which was known as the “onion” structure. Zhang et al. prepared
prepared yolk–shell zeolitic imidazolate framework (ZIF)-8/alginate microcapsules using
yolk–shell zeolitic imidazolate framework (ZIF)-8/alginate microcapsules using microfluidics [57].
microfluidics [57]. Yolk–shell W/O ZIF-8 Pickering emulsions were the template of ZIF-8/alginate
Yolk–shell W/O ZIF-8 Pickering emulsions were the template of ZIF-8/alginate hybrid shell generation,
hybrid shell generation, and the yolks were from phase separation of polyethylene glycol and
and the yolks were from phase separation of polyethylene glycol and acrylamide solution induced
acrylamide solution induced by polymerization. Multi-yolk structures could be obtained by droplet
by polymerization. Multi-yolk structures could be obtained by droplet coalescence. Rhodamine B
coalescence. Rhodamine B controlled release of the materials showed drug carrier potential.
controlled release of the materials showed drug carrier potential.
Based on the phase interface effect, such as the interface reaction and interface self-assembly,
Based on the phase interface effect, such as the interface reaction and interface self-assembly,
hollow microparticles could be synthesized, which is a kind of special, and the most simple, core–
hollow microparticles could be synthesized, which is a kind of special, and the most simple, core–shell
shell microparticle. The interfacial reaction refers to the reaction at the phase interface induced by
microparticle. The interfacial reaction refers to the reaction at the phase interface induced by the
the chemicals in the two phases, and presents a shell-like product. As the reaction progresses, the
chemicals in the two phases, and presents a shell-like product. As the reaction progresses, the increased
increased shell thickness inhibits the diffusion of reactant molecule across the two phases, and ends
the reaction, leaving an empty shell structure. Typical work, such as hollow SiO2 microspheres
synthesized by James Yang and co-workers [58], is shown in Figure 7b. Interface self-assembly is
based on the self-assembly of colloidal particles or molecules at the two phase interfaces. The
Kumacheva group used CO2 as the dispersed phase, and colloidal particle suspension as the
Micromachines 2017, 8, 255 10 of 29

shell thickness inhibits the diffusion of reactant molecule across the two phases, and ends the reaction,
leaving an empty shell structure. Typical work, such as hollow SiO2 microspheres synthesized by
James Yang and co-workers [58], is shown in Figure 7b. Interface self-assembly is based on the
self-assembly of colloidal particles or molecules at the two phase interfaces. The Kumacheva group
used CO2 as the dispersed phase, and colloidal particle suspension as the continuous phase, to form
uniform bubbles in the chip. As CO2 dissolved in the continuous water phase, the bubble volume
and water phase pH decreased, and led to the assembly of colloidal particles at the gas–liquid phase
interface. Microcapsules
Micromachines 2017, 8, 255 with a hollow structure were formed [59], as shown in Figure 7c. 10 of 29

50 μm

100 μm

b Hydrolysis
of TEOS

pH=11.0
Shell
Formation
Wash

c CO2 (g)
Particle
dispersion

100 μm

Figure 7. Core-shell
Figure 7. Core-shell structure
structure microparticles
microparticles prepared
prepared byby phase
phase separation
separation and
and phase
phase interface
interface
effect-based microfluidics.(a)(a)
effect-based microfluidics. Phase
Phase separation
separation process
process of theof the polyethylene
polyethylene glycol diacrylate
glycol diacrylate (PEGDA)
(PEGDA)
core-shell core-shell microparticles;
microparticles; (b) Hollow (b)SiO
Hollow SiO2 microspheres
2 microspheres from interfacial
from interfacial reaction; reaction; (c)
(c) Interface
Interface self-assembly induced hollow structured microcapsules. Reproduced with permission from
self-assembly induced hollow structured microcapsules. Reproduced with permission from [56,58,59].
[56,58,59].
2.1.4. Porous Microparticles
2.1.4. Porous Microparticles
Porous microparticles can be classified into microporous, mesoporous, and macroporous
Porouswith
structures, microparticles can be pore
the corresponding classified
sizes into microporous,
of lower than 2 nm, mesoporous,
from 2 to 50and nm,macroporous
and greater
structures,
than 50 nm, respectively [60,61]. The pores can only exist on the surface of the material andthan
with the corresponding pore sizes of lower than 2 nm, from 2 to 50 nm, and greater can
50
alsonm, respectively
penetrate [60,61].
the entire The Compared
particle. pores can only
with exist on the
the solid surfacesuch
materials, of the material
a porous and can
structure also
greatly
penetrate thespecific
increases the entire particle.
area andCompared
light weight.with the solid materials, such a porous structure greatly
increases the specific area and light weight.
Microspheres with a porous surface can be formed by template occupying and removing, or
Microspheres
modified with smallwith a porouson
size particles surface can be
the particle formed
surface. by and
Yang template occupying
co-workers and removing,
fabricated or
microspheres
modified with small size particles on the particle surface. Yang and co-workers
with hierarchical surface nanopatterns in a microfluidic system [62]. As shown in Figure 8a, the silica fabricated
microspheres with hierarchical
particles in photocurable surface
droplets nanopatterns
protruded in ainterface
across the microfluidic system [62].
as hexagonal As shown
arrays, in
and after
Figure 8a, the silica
polymerization, particles
the silica in photocurable
particles droplets
were selectively protruded
decorated with across the interface as
silver nanoparticles, hexagonal
which could
arrays, and after polymerization, the silica particles were selectively decorated with silver
nanoparticles, which could be used as hot spots for molecular detection based on surface-enhanced
Raman scattering (SERS). Further functions of the microparticles were explored for structural
colored or magnetoresponsive microspheres. In the work of Kumacheva et al. [63], the dispersed
phase was a homogeneous mixture of monomers and solvents, and emulsified as O/W droplets. As
Micromachines 2017, 8, 255 11 of 29

be used as hot spots for molecular detection based on surface-enhanced Raman scattering (SERS).
Further functions of the microparticles were explored for structural colored or magnetoresponsive
microspheres. In the work of Kumacheva et al. [63], the dispersed phase was a homogeneous mixture
of monomers and solvents, and emulsified as O/W droplets. As shown in Figure 8b, after phase
separation induced by photopolymerization and porogen solvent removing, uniform microparticles
were prepared with a network from the surface to the inside. The great specific surface area of the
Micromachines 2017, 8, 255
materials showed their application prospects in the biomedical field. 11 of 29
As shown in Figure 8c, by using flow lithography approach, Doyle et al. combined particle
part with a encoding,
fabrication, unique square macroporous
and probe pattern,
incorporation intowhile the step
a single other[64,65].
part contained a probe molecule
The microparticles had one
with a unique
part with signature,
a unique squarewhich could bepattern,
macroporous used for scanning
while DNA
the other part and proteinsain
contained a single
probe sample.
molecule withIt
was expected to promote the development of low-cost clinical diagnosis.
a unique signature, which could be used for scanning DNA and proteins in a single sample. It was
Qin to
expected and co-workers
promote synthesized
the development biomimetic
of low-cost honeycomb
clinical diagnosis. microparticles based on the
synergistic
Qin andeffect of double
co-workers emulsion
synthesized templates,
biomimetic polymermicroparticles
honeycomb precipitation,based
and oneffervescent salt
the synergistic
decomposition in the microfluidic chip [66]. The materials were composed
effect of double emulsion templates, polymer precipitation, and effervescent salt decomposition in the of an external
nanoporous membrane
microfluidic chip [66]. The and internalwere
materials cavities, and the
composed biological
of an functions asmembrane
external nanoporous microcarriers
and in drug
internal
delivery and cell culture were explored. In the work of Xia et al., as shown in Figure
cavities, and the biological functions as microcarriers in drug delivery and cell culture were explored. 8d [67], porous
microparticles
In the work of Xia with several
et al., micron
as shown holes, 8d
in Figure or [67],
dozens of micron
porous holes, were
microparticles prepared.
with several Different
micron holes,
separated
or dozens layers
of micronof the W/Owere
holes, initial emulsionDifferent
prepared. were selected for W/O/W
separated double
layers of emulsion
the W/O initialgeneration
emulsion
in theselected
were capillary
for coaxial
W/O/Wstructure. After organic
double emulsion solvent
generation volatilization
in the and structure.
capillary coaxial water phaseAfterremoval,
organic
through-pore microparticles
solvent volatilization and water with different
phase removal,pore sizes were microparticles
through-pore achieved. Successful inoculation
with different and
pore sizes
continuous cultivation of fibroblast indicated a better biocompatibility of the
were achieved. Successful inoculation and continuous cultivation of fibroblast indicated a better PLGA microspheres
with macropores.of the PLGA microspheres with macropores.
biocompatibility

Figure
Figure 8.8. Porous
Porous microparticles
microparticles prepared
prepared byby microfluidics.
microfluidics. (a)
(a) Hierarchically-structured
Hierarchically-structured microsphere
microsphere
surface with decorated
surface with decorated particles;
particles; (b)
(b) Microparticles
Microparticles prepared
prepared with
with aa network
network from
from the
the surface
surface to
to the
the
inside from microfluidic O/W droplets; (c) Flow lithography based encoded microparticles;
inside from microfluidic O/W droplets; (c) Flow lithography based encoded microparticles; (d) Cell (d) Cell
carrier
carrier microparticles with controllable
microparticles with controllable pore
pore sizes.
sizes. Reproduced
Reproduced with
with permission
permission from
from [62–65,67].
[62–65,67].

2.1.5.
2.1.5. Composite
Composite Microparticles
Microparticles
Composite
Composite microparticles
microparticles are
are aa kind of material
kind of material with
with different
different compositions,
compositions, and
and functional
functional
characteristics beyond single component particles. Although there is some overlap with
characteristics beyond single component particles. Although there is some overlap with the special the special
shaped or structured
shaped or structuredmaterials,
materials,ininconsideration
considerationthat
that multicomponent
multicomponent would
would play
play a decisive
a decisive rolerole in
in the
the function of materials under some situations, composite microparticles are listed as a separate
function of materials under some situations, composite microparticles are listed as a separate category.
category.
The most straightforward composite materials could be generated by adding different kinds
The most
of materials tostraightforward composite
the dispersion phase, suchmaterials
as dyes, could be generated
nanoparticles, by adding
quantum different
dots and kinds of
biomolecules.
materials to the dispersion phase, such as dyes, nanoparticles, quantum dots and biomolecules.
By polymerization or other reactions, the composite materials are formed with various functions. By
polymerization or other reactions, the composite materials are formed with various
Whitesides et al. blended fluorescent dyes, CdSe QDs, pore-foaming agents, magnetic particles and functions.
Whitesides et al.
liquid crystals blended
into fluorescentglycol
the tripropylene dyes,diacrylate
CdSe QDs, pore-foaming
(TPGDA) agents, magnetic
photopolymer monomerparticles
solutionandfor
liquid crystals into the tripropylene glycol diacrylate (TPGDA) photopolymer monomer solution for
fabrication of the composite microparticles, as shown in Figure 9a [45], which displayed the
diversity and simplicity of additive component manipulation for material synthesis. Silver
nanoparticle loaded chitosan composite microparticles used for antibacterial were also reported, as
shown in Figure 9b [68].
Micromachines 2017, 8, 255 12 of 29

fabrication of the composite microparticles, as shown in Figure 9a [45], which displayed the diversity
and simplicity of additive component manipulation for material synthesis. Silver nanoparticle loaded
Micromachines 2017, 8, 255
chitosan composite 12 of
microparticles used for antibacterial were also reported, as shown in Figure 9b 29
[68].

200 μm

200 μm

Figure 9. Composite microparticles prepared by microfluidics. (a) Microspheres labeled with

Figure 9. Composite microparticles prepared by microfluidics. (a) Microspheres labeled with 4-amino-7-
4-amino-7-nitrobenzo-2-oxa-1,3-diazole (NBD) dye and CdSe quantum dots; (b) Silver nanoparticle–
nitrobenzo-2-oxa-1,3-diazole (NBD) dye and CdSe quantum dots; (b) Silver nanoparticle–chitosan
chitosan composite microparticles assisted by microfluidic synthesis. Reproduced with permission
composite microparticles assisted by microfluidic synthesis. Reproduced with permission from [45,68].
from [45,68].

Janus microparticles
Janus microparticles are are aa typical
typical hybrid
hybrid material.
material. The The two
two parts
parts areare different,
different, and
and thus
thus have
have
multiple properties.
multiple properties. In In the
the microchannel,
microchannel, free free flowing
flowing liquids
liquids have
have laminar
laminar properties
properties andand cancan bebe
changed into droplets under the shear stress. With these two features, Janus
changed into droplets under the shear stress. With these two features, Janus droplets, as shown in droplets, as shown in
Figure 10a,
Figure 10a,could
could be generated,
be generated,and corresponding
and correspondingJanus microparticles were formed
Janus microparticles afterformed
were solidification.
after
Cathala and co-workers induced gelation by diffusion effect and obtained
solidification. Cathala and co-workers induced gelation by diffusion effect and obtained Janus microgels containing
Janus
two different
microgels biopolymers.
containing two In particular,
different biopolymersIncan
biopolymers. be selectively
particular, degradedcan
biopolymers in one be hemisphere
selectively
through enzymatic hydrolysis for the release of a specific inclusion, as
degraded in one hemisphere through enzymatic hydrolysis for the release of a specific inclusion, shown in Figure 10b [69].
as
Based on two-phase Janus microparticle synthesis, the researchers further
shown in Figure 10b [69]. Based on two-phase Janus microparticle synthesis, the researchers further prepared microparticles
with three-phase
prepared or even with
microparticles multiphase, with axisymmetric
three-phase structures.
or even multiphase, Takeuchi
with et al. arranged
axisymmetric structures.the
capillarieset
Takeuchi at al.
a certain anglethe
arranged to capillaries
control the at angular arrangement
a certain angle to of different
control theand simultaneous
angular arrangement laminarof
fluids from these capillary arrays. Under the continuous phase shear action
different and simultaneous laminar fluids from these capillary arrays. Under the continuous phase offered by the centrifugal
force, action
shear the multiphase
offered bylaminar liquid formed
the centrifugal force, multi-component,
the multiphase laminar axial–symmetrical
liquid formeddroplets. After the
multi-component,
axial–symmetrical droplets. After the ionic crosslinking reaction, composite microparticles withand
ionic crosslinking reaction, composite microparticles with the calcium alginate as the body, the
differentalginate
calcium hybrid as compositions
the body, and asdifferent
the compartment, were obtained,
hybrid compositions as shown in Figure
as the compartment, 10c [70].
were obtained,
Cells
as and in
shown magnetic
Figure 10cnanoparticles
[70]. Cells were encapsulated
and magnetic separately
nanoparticles in their
were respectiveseparately
encapsulated unit areas,inwhichtheir
facilitated maintenance of cell viability. Bong et al. utilized microfluidics to synthesize
respective unit areas, which facilitated maintenance of cell viability. Bong et al. utilized microfluidics pH-sensitive,
multimodulated,
to anisotropicmultimodulated,
synthesize pH-sensitive, drug carrier particles, including
anisotropic drugJanus-type, as shown
carrier particles, in Figure
including 10d [71].
Janus-type,
Due
as to theinacidic
shown FigurepH 10dsensibility
[71]. Due of the acidic
to the materials, tumor-selective
pH sensibility drug release
of the materials, could be realized.
tumor-selective drug
The capability of multiple drug encapsulations in the same carrier,
release could be realized. The capability of multiple drug encapsulations in the same and independent release
carrier,of each
and
drug, rendered the potential of synergistic combinatorial cancer treatment.
independent release of each drug, rendered the potential of synergistic combinatorial cancer Besides droplet template
method, phase
treatment. separation
Besides dropleteffect of droplets
template method, canphase
also trigger the formation
separation of the Janus
effect of droplets can structure.
also trigger Weitz
the
formation of the Janus structure. Weitz et al. synthesized Janus microspheres of polyacrylamide and
poly (N-isopropylacrylamide), as shown in Figure 10e [72].
Micromachines 2017, 8, 255 13 of 29

et al. synthesized Janus microspheres of polyacrylamide and poly (N-isopropylacrylamide), as shown

Micromachines
in Figure 10e 2017, 8, 255
[72]. 13 of 29

a b c

50 μm 100 μm

d e

100 μm 200 μm

Figure
Figure 10.
10. Janus
Janus microparticles
microparticles prepared
prepared byby microfluidics.
microfluidics. (a)
(a) Microfluidic
Microfluidic design
design for
for Janus
Janus droplet
droplet
generation;
generation; (b)(b) Biopolymer-based
Biopolymer-based Janus microbeads
Janus microbeads with
with selective selective(c) Multi-compartment
degradation; degradation; (c)
Multi-compartment
particles obtained fromparticles obtained
a multi-barreled from (d)aJanus-type,
capillary; multi-barreled capillary;
pH-responsive (d) via
particles Janus-type,
stop-flow
pH-responsive particles viaand
lithography; (e) Aggregation stop-flow
compactionlithography; (e) Aggregation and compaction
of the Poly(N-isopropylacrylamide) of the
(PNIPAm) microgels.
Poly(N-isopropylacrylamide)
Reproduced with permission from (PNIPAm) microgels. Reproduced with permission from [69–72].
[69–72].

2.2.
2.2. Particulate
Particulate Biomaterials
Biomaterials at
at the
the Nanoscale
Nanoscale
Nanoparticles
Nanoparticles havehave drawn
drawn significant
significant attention,
attention, due due to to their
their unique
unique andand excellent
excellent properties.
properties.
Compared to the batch synthesis process, the microfluidic
Compared to the batch synthesis process, the microfluidic platform can provide platform can provide precise
precisecontrol of
control
the temperature,
of the temperature,concentration,
concentration, and mixing
and mixingprocess for thefor
process synthesis of nanoparticles,
the synthesis thus, the thus,
of nanoparticles, size,
size distribution, morphology, reproducibility, and throughput of nanoparticles,
the size, size distribution, morphology, reproducibility, and throughput of nanoparticles, could be could be effectively
regulated
effectively [73]. The [73].
regulated single-phase flow, multiphase
The single-phase flow, multiphase flow, flow,
multi-step, andand
multi-step, coupling
couplingsystem
system in in
microfluidic
microfluidic devices
devices areare established
established forfor the
the stable
stable microreactor
microreactor conditions
conditions [74].
[74]. The
The synthesis
synthesis of of the
the
particulate
particulate biomaterials
biomaterials at at the
the nanoscale
nanoscale include
include metal
metal nanoparticles,
nanoparticles, oxide oxide nanoparticles,
nanoparticles, polymer
polymer
nanoparticles,
nanoparticles, hybrid
hybrid nanoparticles,
nanoparticles, and and quantum
quantum dots dots [75,76],
[75,76], which
which are
are applied
applied in in bioimaging,
bioimaging,
biosensing,
biosensing, drug
drug delivery,
delivery, diagnosis,
diagnosis, andand therapy.
therapy. Hassan’s
Hassan’s group group employed
employed 3D 3D coaxial
coaxial flow
flow device
device
with a heat supply to fabricate magnetic nanoparticles [77]. Utilizing
with a heat supply to fabricate magnetic nanoparticles [77]. Utilizing the multistep microfluidic the multistep microfluidic
device
device with
with three
three separated
separated microreactors,
microreactors, fluorescent
fluorescent silica-coated
silica-coated magnetic
magnetic nanoparticles
nanoparticles were were
synthesized, which could be used for magnetic resonance imaging (MRI)
synthesized, which could be used for magnetic resonance imaging (MRI) and fluorescence imaging [78]. and fluorescence imaging
[78]. For molecular
For molecular imaging,imaging, Liu[79]
Liu et al. et prepared
al. [79] the prepared the supramolecular
supramolecular nanoparticles nanoparticles
with controllablewith
controllable
size and surfacesizechemistry
and surface chemistry in a polydimethylsiloxane
in a polydimethylsiloxane (PDMS) microfluidic(PDMS) microfluidic
chip. Evaluation chip.
of cellular
Evaluation of cellular
uptake efficiency of theuptake efficiencydisplayed
nanoparticles of the nanoparticles
the capability displayed the capability in bioimaging.
in bioimaging.
Metal
Metal nanoparticles
nanoparticles used used forfor biosensing,
biosensing,such suchasassilver
silver[80]
[80]andandgold
gold [81],
[81], were
were first
first fabricated
fabricated by
by microfluidics in single flows, and with more rapid reaction
microfluidics in single flows, and with more rapid reaction speeds and narrower size distributions speeds and narrower size
distributions
compared tocompared
those from to those from bulk synthesis.
bulk synthesis. SadAbadiSadAbadi et al. integrated
et al. integrated in-situin-situ synthesized
synthesized gold
gold nanoparticles
nanoparticles into into microdevices,
microdevices, which
which served
served as as
a alocalized
localizedsurface
surfaceplasmon
plasmon resonance
resonance based
based
biosensor
biosensor forfor protein
protein andand polypeptide
polypeptide detection,
detection, as as shown
shown in in Figure
Figure 11a
11a [82].
[82]. The
The high
high sensitivity
sensitivity
and
and extreme
extremelowlowdetect
detect limits of the
limits detection
of the detectionsystem suggested
system clinical
suggested application
clinical potential.
application Using
potential.
segmented flow microfluidic
Using segmented systems, Knauer
flow microfluidic systems, and co-workers
Knauer and prepared
co-workers noble metal nanoparticles
prepared noble metal
with core-shellwith
nanoparticles andcore-shell
multi-shell andstructure
multi-shell [83]. The great
structure [83].change
The great in change
optical incharacteristics of these
optical characteristics
nanoparticles showed promising
of these nanoparticles potentialpotential
showed promising for plasmonic applications.
for plasmonic applications.
Biodegradable polymeric nanoparticles with narrow size distribution, high reproducibility, and
encapsulation efficiency, are ideal drug and nucleic acid delivery vehicles. Farokhzad’s group
prepared dual drug loaded polymeric nanoparticles in the microfluidic device, as shown in Figure
11b [84], by self-assembly of docetaxel, prodrug poly lactic acid (PLA)–Pt(IV), and block polymer
polyethylene glycol–poly(lactic-co-glycolic acid) (PEG–PLGA), and subsequent modification of the
targeting ligand. Furthermore, they synthesized 45 kinds PEG–PLGA nanoparticles via optimization
Micromachines 2017, 8, 255 14 of 29

Biodegradable polymeric nanoparticles with narrow size distribution, high reproducibility,

and encapsulation efficiency, are ideal drug and nucleic acid delivery vehicles. Farokhzad’s group
prepared dual drug loaded polymeric nanoparticles in the microfluidic device, as shown in
Micromachines 2017, 8, 255 14 of 29
Figure 11b [84], by self-assembly of docetaxel, prodrug poly lactic acid (PLA)–Pt(IV), and block polymer
polyethyleneusing
experiments glycol–poly(lactic-co-glycolic acid) (PEG–PLGA),
multiple precursors [85]. Lipid–polymer and subsequent
nanoparticles for drugmodification of the
delivery included
targeting ligand.
nanoparticles withFurthermore, they synthesized
core–shell structure 45 kinds or
[86], monolayer PEG–PLGA
bilayer ofnanoparticles via optimization
lipid shells [87], and hybrid
experiments using multiple precursors [85]. Lipid–polymer nanoparticles for drug
Janus structure [88]. Leong et al. used an emulsion-based microfluidic system to prepare polymer– delivery included
nucleic acid nanocomplexes
nanoparticles with improved
with core–shell structure nonviralorgene
[86], monolayer transfer
bilayer capability
of lipid [89],
shells [87], andas shown
hybrid in
Janus
Figure 11c. Besides, the small interfering RNA (siRNA) delivery with high loading efficiency and
structure [88]. Leong et al. used an emulsion-based microfluidic system to prepare polymer–nucleic
enhanced stability [90],
acid nanocomplexes oligodeoxynucleotide
with improved nonviral gene delivery
transfer with improved
capability [89], ascellular
shown in uptake
Figureand
11c.
immunostimulatory responses RNA
Besides, the small interfering [91], the co-delivery
(siRNA) of hydrophilic
delivery drug and
with high loading siRNA [92]
efficiency and with high
enhanced
loading efficiency, and co-delivery of gene and proteins [93], would be achieved by microfluidic
stability [90], oligodeoxynucleotide delivery with improved cellular uptake and immunostimulatory
fabricated
responses nanoparticles.
[91], the co-delivery of hydrophilic drug and siRNA [92] with high loading efficiency,
For diagnosis
and co-delivery and therapy
of gene applications,
and proteins [93], wouldusing static micromixer–coaxial
be achieved electrospray,
by microfluidic fabricated Lee and
nanoparticles.
co-workers one-step synthesized theranostic lipoplexes, as shown in Figure 11d [94]. The lipoplexes
For diagnosis and therapy applications, using static micromixer–coaxial electrospray, Lee and
incorporated with QDs,
co-workers one-step Cy5-labeled
synthesized antisenselipoplexes,
theranostic oligodeoxynucleotides, as the model
as shown in Figure imaging
11d [94]. reagent,
The lipoplexes
and treatment drug, respectively. Efficient delivery into cancer cells, and down-regulated
incorporated with QDs, Cy5-labeled antisense oligodeoxynucleotides, as the model imaging reagent,
cancer-related gene expression, revealed the future cancer treatment potential.
and treatment drug, respectively. Efficient delivery into cancer cells, and down-regulated cancer-related
gene expression, revealed the future cancer treatment potential.

a b Polymers + drugs
Hydrodynamic Flow in Acetonitrile
Focusing Water

Nanoparticles

Nanoprecipitation

c d Therapeutic
reagent
Imaging
reagent

16 mixing
units

50 μm
3 kV

Figure
Figure 11.
11.Nanoparticles
Nanoparticlesprepared
preparedby by
microfluidics. (a) In(a)
microfluidics. situ
Insynthesized gold nanoparticles
situ synthesized in the
gold nanoparticles
in the microdevices; (b) Dual drug-loaded polymeric nanoparticles by microfluidic
microdevices; (b) Dual drug-loaded polymeric nanoparticles by microfluidic self-assembly; self-assembly;
(c)
(c) Microfluidic
Microfluidic preparation
preparation of of polymer-nucleicacid
polymer-nucleic acidnanocomplexes;
nanocomplexes; (d) (d) Theranostic
Theranostic lipoplexes
lipoplexes
synthesized from
synthesized from microfluidic
microfluidic coaxial
coaxial electrospray.
electrospray. Reproduced
Reproduced with with permission
permission from
from [82,84,89,94].
[82,84,89,94].

ItIt is
is noted
noted that
that differences
differences in in operating
operating principles
principles of
of flow-focusing
flow-focusing methods
methods do do exist
exist among
among
various preparations. For microparticle synthesis, flow-focusing geometries
various preparations. For microparticle synthesis, flow-focusing geometries are mainly designedare mainly designed for
uniform
for uniform droplet generation,
droplet which
generation, are used
which for microparticle
are used templates.
for microparticle Reaction
templates. or changes
Reaction in the
or changes
droplets or on the
in the droplets or on droplet surfacesurface
the droplet induceinduce
the generation
the generationof theofmicroparticles, whilewhile
the microparticles, for the
for
nanoparticles,
the nanoparticles, flow-focusing
flow-focusinggeometries could
geometries be used
could be usedto to
generate droplet
generate reactors
droplet reactorsor or co-flow
co-flow
reactors
reactors [84,95].
[84,95]. For
For aa given
given set
set of
of conditions,
conditions, such
such as
as the
the channel
channel dimension,
dimension, channel
channel modification
modification
and
and velocity,
velocity, andand stable
stable co-flows
co-flows could
could be
be generated
generated under
under flow-focusing
flow-focusing designs,
designs, as
as the
the reactor
reactor for
for
nanoparticle
nanoparticle synthesis.
synthesis. Among
Amongflow-focusing
flow-focusing designs
designs for
for nanoparticle
nanoparticle synthesis,
synthesis, 3D
3D hydrodynamic
hydrodynamic
focusing
focusing technique
technique requires
requires both
both horizontal
horizontal and
and vertical
vertical focusing
focusing ofof the
the sample
sample flows,
flows, andand can
can
enhance the uniform mixing by the reduced diffusion length via a microfluidic
enhance the uniform mixing by the reduced diffusion length via a microfluidic channel, compared channel, compared
to the 2D hydrodynamic focusing, where the central flow is only focused in the horizontal plane.
For example, shape-controlled tetrathiafulvalene–Au hybrid materials, and polyplexes with high
uniformity and improved biological performance, could be synthesized by 3D hydrodynamic
focusing in straightforward processes [96,97].

3. Fibrous Biomaterials Synthesis and Applications

Micromachines 2017, 8, 255 15 of 29

to the 2D hydrodynamic focusing, where the central flow is only focused in the horizontal plane.
For example, shape-controlled tetrathiafulvalene–Au hybrid materials, and polyplexes with high
uniformity and improved biological performance, could be synthesized by 3D hydrodynamic focusing
in straightforward
Micromachines processes [96,97].
2017, 8, 255 15 of 29

3. Fibrous Biomaterials
Microfluidic Synthesis
spinning and Applications
technology is used as a new technology for synthesizing micro/nano
fibrous
Microfluidic spinning technology is structures,
materials with different shapes, used as a newand technology
components,forwhich have been
synthesizing used as
micro/nano
scaffolds,
fibrous with the
materials withcapability of physiological
different shapes, structures,microenvironment
and components, which mimicking,
have beencellused
encapsulation
as scaffolds,
extended to aspects of cell immobilization, co-culture, immunoprotection,
with the capability of physiological microenvironment mimicking, cell encapsulation extended to and microvascular
construction
aspects of cell[98]. Except for applications
immobilization, co-culture,inimmunoprotection,
tissue engineering, andcontrolled drug delivery
microvascular could also
construction [98].
be realized
Except by microfluidic
for applications fibrousengineering,
in tissue materials [98]. Microfluidic
controlled drugspinning
deliveryoffers
couldadvantages such asby
also be realized
the mild fabrication
microfluidic environment,
fibrous materials uniformity in
[98]. Microfluidic the fiber
spinning sizeadvantages
offers and shape, such
and as
thethe
ability
mild to handle
fabrication
single fibers. Microfluidic microfiber synthesis generally adopts the chip design and
environment, uniformity in the fiber size and shape, and the ability to handle single fibers. Microfluidic manipulation
shown in Figure
microfiber synthesis 12,generally
which is adopts
analogous to droplet
the chip designmicrofluidic chips. shown
and manipulation That is,inby performing
Figure theis
12, which
dimension optimization, channel modification, and velocity adjustment to the
analogous to droplet microfluidic chips. That is, by performing the dimension optimization, channeldroplet microfluidic
chip, stable co-flows
modification, could be
and velocity generated,towhich
adjustment are used
the droplet as microfiber
microfluidic templates.
chip, The principles
stable co-flows couldofbe
generated, which are used as microfiber templates. The principles of fibrous material curingcuring
fibrous material curing are the same as in Section 2. Based on the above technology and are the
principle,
same as in microfiber engineered
Section 2. Based on thefrom
abovemicrofluidics
technology and can curing
be divided into solid
principle, fiber, engineered
microfiber special shaped
from
fiber, bamboocan
microfluidics structure fiber,into
be divided core–shell fiber,
solid fiber, porous
special fiber, fiber,
shaped and multi-component
bamboo structurefiber,fiber,which are
core–shell
also according to the material shape, structure, and composition. Whilst nanofiber synthesis can be
fiber, porous fiber, and multi-component fiber, which are also according to the material shape, structure,
achieved by combining the microfluidic and traditional electrospinning. Besides, special reaction
and composition. Whilst nanofiber synthesis can be achieved by combining the microfluidic and
systems are also available.
traditional electrospinning. Besides, special reaction systems are also available.

a b Continuous
c
Dispersed Continuous phase
phase
phase

Continuous phase Dispersed

phase Dispersed phase

Figure 12.
Figure 12. Microfluidic
Microfluidic technologies
technologies for
for the
the synthesis
synthesis of
of fibrous
fibrous materials.
materials.The
Theprinciples
principlesand
andchip
chip
designswith
designs with different
different flowflow regimes
regimes for co-flow
for stable stable generation,
co-flow generation, including (a);
including T-junction T-junction (a);
flow-focusing
flow-focusing
(b); and coaxial(b);
(c) and coaxialchip.
structured (c) structured chip.

3.1. Fibrous
3.1. Fibrous Biomaterials
Biomaterials at
at Micro-Scale
Micro-Scale
Kangand
Kang andco-workers
co-workersrealized
realized uniform
uniform carving
carving microgroove
microgroove patterns
patterns on theonsurface
the surface of flat
of flat calcium
calcium alginate microfibers by designing a special microstructure in the chip,
alginate microfibers by designing a special microstructure in the chip, as indicated in Figure 13a. as indicated in Figure
13a. structural
This This structural material
material can becanused
be used as a novel
as a novel kindkind of scaffold,
of scaffold, to regulate
to regulate the arrangement
the arrangement of
of cells
cells
in in tissue
tissue engineering
engineering [99]. [99].
Hwang Hwanget al.et synthesized
al. synthesizedPLGA PLGA microfibers
microfibers with
with diametersfrom
diameters fromtentento
to hundreds
hundreds of microns
of microns in in
thethe microfluidicchip.
microfluidic chip.Using
Using different
different fibers,
fibers, they
they found
foundthat
thatthe
thecellular
cellular
orientation could be regulated, which can be applied in microstructure design and constructioninin
orientation could be regulated, which can be applied in microstructure design and construction
regenerative medicine
regenerative medicine and
and tissue
tissue engineering
engineering [100].
[100]. Lee
Lee etetal.
al.developed
developedmicrofiber
microfiberspinning
spinningfor forthe
the
in situ construction of 3D fibrous scaffolds on the same microfluidic device.
in situ construction of 3D fibrous scaffolds on the same microfluidic device. Primary hepatocytes Primary hepatocytes
wereencapsulated,
were encapsulated, and
and maintained
maintained highhigh viability
viability in
in the
the fibrous
fibrous scaffold
scaffoldover
overseven
sevendays.
days.During
Duringthe the
scaffold formation, the soft alginate fibers and encapsulated cells were both
scaffold formation, the soft alginate fibers and encapsulated cells were both without damage [101]. without damage [101].
Shietetal.
Shi al.developed
developedaacell-responsive
cell-responsivemethacrylamide-modified
methacrylamide-modifiedgelatin gelatin(GelMA)
(GelMA)microfiber
microfiberbased
based on
on microfluidic spinning. This fiber possessed microgroove surfaces, and could
microfluidic spinning. This fiber possessed microgroove surfaces, and could be used for cell scaffold be used for cell
scaffold and cell encapsulation, simultaneously. These microstructured fibers could be used as
and cell encapsulation, simultaneously. These microstructured fibers could be used as potential
potential templates for the production of fiber shaped tissues [102]. Yu and co-workers fabricated
bamboo like micron-scale bionic material based on droplet microfluidics and wet spinning [103]. As
shown in Figure 13b, the fibrous materials were about 100–200 μm in diameter, and can be tens of
meters in length. The bamboo pole was natural hydrogel with good biocompatibility, and the
bamboo joint was spherical material in an orderly arrangement, which could be microdroplets or
Micromachines 2017, 8, 255 16 of 29

templates for the production of fiber shaped tissues [102]. Yu and co-workers fabricated bamboo like
micron-scale bionic material based on droplet microfluidics and wet spinning [103]. As shown in
Figure 13b, the fibrous materials were about 100–200 µm in diameter, and can be tens of meters in
length. The bamboo pole was natural hydrogel with good biocompatibility, and the bamboo joint
Micromachines
was spherical 8, 255
2017,material 16 of 29
in an orderly arrangement, which could be microdroplets or multicellular
microspheres. The materials could be used as functional carriers of cells and biomolecules. In the
biomolecules. In the work of Kang et al., tunable physiochemical coding systems in the fiber were
work of Kang et al., tunable physiochemical coding systems in the fiber were generated by a novel
generated by a novel microfluidic spinning method [104]. The capability of coding diverse materials
microfluidic spinning method [104]. The capability of coding diverse materials could be used for the
could be used for the spatiotemporally programmed loading and release of cells or drugs. Ahn et al.
spatiotemporally programmed loading and release of cells or drugs. Ahn et al. developed antibiotic
developed antibiotic alginate fiber as natural polymer-based drug carriers in the microfluidic
alginate fiber as natural polymer-based drug carriers in the microfluidic spinning system, as indicated
spinning system, as indicated in Figure 13c [105]. The materials were with high drug encapsulation
in Figure 13c [105]. The materials were with high drug encapsulation efficiency, and a distinctively
efficiency, and a distinctively delayed degradation profile. The densely packed structure and
delayed degradation profile. The densely packed structure and enhanced drug loading efficiency of
enhanced drug loading efficiency of the fiber were derived from the dehydration effect of polarity
the fiber were derived from the dehydration effect of polarity isopropyl alcohol in the sheath flow.
isopropyl alcohol in the sheath flow. In vivo infected wound healing showed the application
In vivo infected wound healing showed the application potential. Chu and co-workers reported
potential. Chu and co-workers reported chitosan microfibers synthesized from microfluidics [106].
chitosan microfibers synthesized from microfluidics [106]. The materials were with controllable
The materials were with controllable internal structures, from tubular to peapod-like structures,
internal structures, from tubular to peapod-like structures, which could be used for transporting fluid
which could be used for transporting fluid and encapsulation of multiple drugs, respectively.
and encapsulation of multiple drugs, respectively.

a b

50 μm 50 μm
Smooth Grooved
50 μm
PLGA +
Fluorescein
50 μm 100 μm
Grooved

Figure
Figure 13.
13. Micro-scale
Micro-scalefibers
fibers prepared
prepared by
by microfluidics.
microfluidics. (a)(a)Flat
Flat alginate
alginate fibers
fibers with
with groove
groove
microstructures;
microstructures; (b)
(b) Biomimetic
Biomimetic bamboo-like
bamboo-like hybrid
hybrid microfibers;
microfibers; (c)
(c) Fibrous
Fibrous alginate
alginate carrier
carrier from
from
microfluidic
microfluidic spinning.
spinning. Reproduced
Reproduced with
with permission
permission from
from [99,103,105].
[99,103,105].

3.2.
3.2. Fibrous
FibrousBiomaterials
Biomaterials at
at the
the Nanoscale
Nanoscale
The
The diameters
diametersof of fibers
fibers fabricated
fabricatedby by microfluidics
microfluidicsrangerangefrom
from aa few
few micrometers
micrometers to to hundreds
hundreds
of
of micrometers [98]. Due to the difficulties in fabricating nanoscale microfluidic channels, fibrous
micrometers [98]. Due to the difficulties in fabricating nanoscale microfluidic channels, fibrous
materials
materialsscaling
scalingdown
downtotonanometer
nanometerare arechallenging
challenging [107].
[107].However,
However, there hashas
there stillstill
beenbeenattempts to
attempts
synthesize fibrous biomaterials at the nanoscale. Lee et al. prepared nano-scaled alginate
to synthesize fibrous biomaterials at the nanoscale. Lee et al. prepared nano-scaled alginate fibers fibers using
isopropyl alcoholalcohol
using isopropyl as the as sheath flow inflow
the sheath the in
microfluidic system,
the microfluidic as shown
system, in Figure
as shown 14a [108].
in Figure The
14a [108].
alginate nanostrands
The alginate were were
nanostrands formed by isopropyl
formed alcohol-induced
by isopropyl dehydration.
alcohol-induced Thinner
dehydration. fibers could
Thinner fibers
be generated
could from from
be generated these these
nanostrands
nanostrandsunder the the
under shear force
shear in in
force thethe
microchannel
microchannelinto intoaacompact
compact
structure. Besides,asasshown
structure. Besides, shown in Figure
in Figure 14b, 14b,
ZhangZhang et al. combined
et al. combined the microfluidic
the microfluidic gradient
gradient generation
generation and electricspinning,
and electricspinning, to synthesize to synthesize
the nanofiberthe nanofiber
scaffold with scaffold with concentration
multiple multiple concentration
gradients
gradients of large molecular proteins, small molecule drugs, and
of large molecular proteins, small molecule drugs, and nanoparticles [109]. The nanoparticles [109]. The spinning
spinning scaffold
scaffold containing the dexamethasone gradient was used to induce osteogenesis and adipogenic
differentiation of mesenchymal stem cells.
Micromachines 2017, 8, 255 17 of 29

containing the dexamethasone gradient was used to induce osteogenesis and adipogenic differentiation
Micromachines 2017, 8, 255 17 of 29
of mesenchymal stem cells.

Figure
Figure 14.
14. Nano-scale
Nano-scale fibers
fibers prepared
prepared by
by microfluidics. (a) Micro/nanometer-scale
microfluidics. (a) Micro/nanometer-scale fiber
fiber with
with ordered
ordered
structures;
structures; (b) Gradient electrospinning
(b) Gradient electrospinning nanofibers
nanofibers assisted
assisted by
by microfluidics.
microfluidics. Reproduced
Reproduced with
with
permission from [108,109].
permission from [108,109].

4.
4. Sheet
Sheet Biomaterials
Biomaterials Synthesis
Synthesis and
and Applications
Applications

Günther
Günther et al. developed
et al. developed aa digital
digital controlled
controlled “textile”
“textile” technology
technology for for mosaic
mosaic hydrogel
hydrogel sheetsheet
synthesis [110]. The
synthesis [110]. Thehighly
highlyintegrated
integrated fluids
fluids were
were regulated
regulated throughthrough particular
particular programs,
programs, and a and
series a
series of cell patterns were constructed, as indicated in Figure 15a. This work realized high-density
of cell patterns were constructed, as indicated in Figure 15a. This work realized high-density cell culture
cell culture
in the in the unsupported
unsupported soft material, soft material,
and offered anand
ideaoffered an idea
for research for relationship
on the research onbetween
the relationship
cell–cell,
between cell–cell,
cell–matrix, and 3D cell–matrix,
functional and 3D functional
organization organization
construction in theconstruction
physiologicalinmicroenvironment.
the physiological
microenvironment.
Hanagata and co-workers Hanagata andsynthesized
in situ co-workersmicrogroove
in situ synthesized
silica nanotubemicrogroove
membranes silicainnanotube
a Teflon
membranes
microfluidicin chipa Teflon
[111]. microfluidic
Sustained releasechip [111].
of bone Sustained release of
morphogenetic bone 2morphogenetic
protein protein 2
(BMP-2) and osteoblast
(BMP-2) and osteoblast
differentiation induction differentiation
could be realized induction could be realized
by the membranes, by the
indicating membranes,
their indicating
multiple potentials as
their multipleand
cell scaffolds potentials as cellfor
drug carriers scaffolds and drug carriers
tissue regeneration. Seki etforal.tissue regeneration.
utilized micronozzleSeki et al.
device utilized
patterned
micronozzle
hydrogel sheet device patterned
for the hydrogel
high-density sheet for the
3D co-culture high-densityand
of hepatocytes 3D fibroblasts
co-culture of hepatocytes
[112]. As indicated and
fibroblasts [112].
in Figure 15b, As indicated
HepG2 and 3T3inwere
Figure 15b, HepG2
co-cultured forand 3T3days
seven werein co-cultured
the uniform for film
seven ofdays
100 µm in thein
uniform
thicknessfilm
andofseveral
100 μm in thickness
millimeters in and
width,several millimeters
and formed in width, andsimilar
micro-organoids formedtomicro-organoids
the structure of
similar to the structure
in vivo hepatic of in vivo previous
cord. To overcome hepatic cord. To overcome
limitations previous
in the scalable limitations
formation in the scalable
of polymeric films,
formation
such as lack ofofpolymeric
pore size,films,
shape,such as lack control,
and surface of pore Elsayed
size, shape,et al.and surface acontrol,
introduced Elsayed
microfluidic et al.
junction
introduced a microfluidic
and coarse capillaries junction ordered
to synthesize and coarse capillaries
porous to synthesize
films [113]. ordered porousbubbles
Highly monodispersed films [113].
were
Highly monodispersed
firstly generated bubbles
in T-junction were firstly
microfluidic generated
capillaries. By in T-junction
regulating themicrofluidic capillaries.and
polymer concentration By
regulating the polymer
the surfactant concentration
type, microbubbles withand the surfactant
controllable poretype,
size microbubbles
were collectedwith controllable
on slides to formpore the
size were collected on slides to form the porous structures. Selective nanoparticles could be
embedded into the films for extensive application in biomedicine.
Micromachines 2017, 8, 255 18 of 29

porous structures. Selective nanoparticles could be embedded into the films for extensive application
Micromachines 2017, 8, 255
in biomedicine. 18 of 29

a Gear Pump Focusing

Fluid Imaging b
Drum

Base
Biopolymer

Microfluidic
Device

Figure 15. Sheet materials prepared by microfluidics. (a) Mosaic hydrogels sheets, scale bars are all
Figure 15. Sheet materials prepared by microfluidics. (a) Mosaic hydrogels sheets, scale bars are all
500 μm; (b) Micronozzle device patterned hydrogel sheet. Reproduced with permission from
500 µm; (b) Micronozzle device patterned hydrogel sheet. Reproduced with permission from [110,112].
[110,112].

5. Construct Forms of Biomaterials Synthesis and Applications

5. Construct Forms of Biomaterials Synthesis and Applications
Hydrogel assembly is an effective way to construct soft materials for tissue engineering.
Hydrogel assembly is an effective way to construct soft materials for tissue engineering.
Sometimes, due to the poor operation controllability of hydrogels, it remains a challenge to fabricate
Sometimes, due to the poor operation controllability of hydrogels, it remains a challenge to fabricate
morphologically accurate structures in vitro. Utilizing microfluidic platforms, assembled microgel
morphologically accurate structures in vitro. Utilizing microfluidic platforms, assembled microgel
material containing different types of cells could be fabricated [114]. As indicated in Figure 16a, in
material containing different types of cells could be fabricated [114]. As indicated in Figure 16a, in a
a bottom-up fabrication manner, 3D cell-laden microfluidic constructs were synthesized by Li and
bottom-up fabrication manner, 3D cell-laden microfluidic constructs were synthesized by Li and
co-workers. By rolling monolayer cell-loaded scaffolds, obtained microfluidic channels and primary
co-workers. By rolling monolayer cell-loaded scaffolds, obtained microfluidic channels and primary
hepatocytes were uniformly distributed in 3D, which could be potentially used to engineer 3D
hepatocytes were uniformly distributed in 3D, which could be potentially used to engineer 3D
vascularized liver organs [115]. Qin et al. prepared collagen constructs with tunable geometries
vascularized liver organs [115]. Qin et al. prepared collagen constructs with tunable geometries
through a membrane-templated microdevice, which could be used for 3D construct self-assembly [116],
through a membrane-templated microdevice, which could be used for 3D construct self-assembly
as indicated in Figure 16b. By encapsulating tissue-specific cells into the constructs, functional
[116], as indicated in Figure 16b. By encapsulating tissue-specific cells into the constructs, functional
microtissues were generated. Derived by the combined effects of cell–cell and cell–matrix interactions,
microtissues were generated. Derived by the combined effects of cell–cell and cell–matrix
cell-laden constructs spontaneous self-assembled into tissue-like constructs. The maximum lengths
interactions, cell-laden constructs spontaneous self-assembled into tissue-like constructs. The
of tubular constructs, 6 mm, could be achieved by this method. Seki et al. prepared vascular tissues
maximum lengths of tubular constructs, 6 mm, could be achieved by this method. Seki et al.
with multilayered, branched, and thick structures, using a microfluidic device made of agarose
prepared vascular tissues with multilayered, branched, and thick structures, using a microfluidic
hydrogel [117]. Smooth muscle cell embedding Ca–alginate hydrogel layer was generated due to the
device made of agarose hydrogel [117]. Smooth muscle cell embedding Ca–alginate hydrogel layer
reaction between Ca2+ diffused from the agarose channel body and introduced alginate–cell solution.
was generated due to the reaction between Ca2+ diffused from the agarose channel body and
Continuous culture of endothelial cells on this hydrogel layer induced the formation of vascular tissues.
introduced alginate–cell solution. Continuous culture of endothelial cells on this hydrogel layer
More importantly, by removing the agarose hydrogel plates, the vascular tissues could be separated as
induced the formation of vascular tissues. More importantly, by removing the agarose hydrogel
independent constructs, which enabled vascular tissue based biomedical applications.
plates, the vascular tissues could be separated as independent constructs, which enabled vascular
There have been several works on the assembly of microfluidic spun microfibers into 3D constructs
tissue based biomedical applications.
via reeling, weaving, or direct writing [98]. With accumulated spun microfibers, the dimensions
There have been several works on the assembly of microfluidic spun microfibers into 3D
of 3D materials in at least two directions could be at millimeter scale. Fukuda et al. adopted a
constructs via reeling, weaving, or direct writing [98]. With accumulated spun microfibers, the
magnetic-driven strategy to assemble the cell-laden alginate hydrogel microfibers into macroscopic
dimensions of 3D materials in at least two directions could be at millimeter scale. Fukuda et al.
cellular structures with high cell viability [118]. Lee et al. constructed an artificial liver system
adopted a magnetic-driven strategy to assemble the cell-laden alginate hydrogel microfibers into
on the chip by continuously reeling the HepG2-cultured chitosan microfibers [119]. Onoe et al.
macroscopic cellular structures with high cell viability [118]. Lee et al. constructed an artificial liver
flexibly handled the cell-laden microfibers using a thin tube and fluid flow, which was revealing for
system on the chip by continuously reeling the HepG2-cultured chitosan microfibers [119]. Onoe et
fiber-shaped cellular construct, as shown in Figure 16c [120]. Juncker et al. used a microfluidic direct
al. flexibly handled the cell-laden microfibers using a thin tube and fluid flow, which was revealing
writer to create 3D cell-laden hydrogel constructs under computer-controlled operating systems [121].
for fiber-shaped cellular construct, as shown in Figure 16c [120]. Juncker et al. used a microfluidic
These layer-by-layer assembled 3D structures were with openings permitting media exchange, which
direct writer to create 3D cell-laden hydrogel constructs under computer-controlled operating
was critical for the encapsulated cells. Zhang et al. adopted cell-laden fibers with hierarchically
systems [121]. These layer-by-layer assembled 3D structures were with openings permitting media
exchange, which was critical for the encapsulated cells. Zhang et al. adopted cell-laden fibers with
hierarchically organized architecture for the synthesis of tissue-like constructs, in which the
mechanical properties and biological activity could be modulated by the use of photo-cross-linkable
methacrylated alginate as the raw materials [122], as shown in Figure 16d.
Micromachines 2017, 8, 255 19 of 29

organized architecture for the synthesis of tissue-like constructs, in which the mechanical properties
and biological activity could be modulated by the use of photo-cross-linkable methacrylated alginate
Micromachines 2017, 8, 255 [122], as shown in Figure 16d.
as the raw materials 19 of 29

a b
1 cm

100 μm
1 cm

1 cm 200 μm
20 μm 10 μm

c d

1 mm

1 mm

1 mm 1 mm

Figure 16. Construct forms of materials prepared by microfluidics. (a) Bottom-up fabrication of 3D
Figure 16. Construct forms of materials prepared by microfluidics. (a) Bottom-up fabrication of 3D
microfluidic cell-ladenconstructs;
microfluidic cell-laden constructs; (b) Collagen
(b) Collagen building
building constructs
constructs for self-assembly
for self-assembly of 3D
of 3D microtissues;
microtissues; (c) Microfiber-based
(c) Microfiber-based assembly of 3D assembly of 3D cellular
macroscopic macroscopic cellular(d)structures;
structures; Constructs (d)made
Constructs
by the
made by the automated assembly of in situ formed microfibers. Reproduced with permission
automated assembly of in situ formed microfibers. Reproduced with permission from [115,116,120,122]. from
[115,116,120,122].

6. Summaries and Outlook

6. Summaries and Outlook
In the last decade, there has been a rapid increase in the number and quality of reports on
In the last decade, there has been a rapid increase in the number and quality of reports on
functional biomaterial synthesis based on microfluidic chip platforms. With the development of
functional biomaterial synthesis based on microfluidic chip platforms. With the development of
microfluidic preparation technology and related detection methods, biomaterials engineered from
microfluidic preparation technology and related detection methods, biomaterials engineered from
microfluidic technologies have experienced transition from simplicity to complexity in material
microfluidic technologies have experienced transition from simplicity to complexity in material
structure, and simplification to diversity in material function, including the entire micro/nano-material
structure, and simplification to diversity in material function, including the entire
dimension from 0D particulate materials, 1D fibrous materials, 2D sheet materials, to 3D construct
micro/nano-material dimension from 0D particulate materials, 1D fibrous materials, 2D sheet
forms of materials, which largely compensates for the lack of conventional synthesis methods. Based
materials, to 3D construct forms of materials, which largely compensates for the lack of conventional
on the flexibility and integration of microfluidic manipulation, preparation of functional particulate
synthesis methods. Based on the flexibility and integration of microfluidic manipulation,
materials has extended from the original single emulsion droplet method to complex multiple emulsion
preparation of functional particulate materials has extended from the original single emulsion
droplet method, micro-channel limiting, flow lithography, and so on. The resultant particulate
droplet method to complex multiple emulsion droplet method, micro-channel limiting, flow
materials have developed from simple solid microparticles to specially shaped microparticles, hollow
lithography, and so on. The resultant particulate materials have developed from simple solid
particles, core–shell structured particles, and hybrid particles. A series of novel nanoparticles
microparticles to specially shaped microparticles, hollow particles, core–shell structured particles,
has also been developed in the microfluidic system. For microfiber materials synthesized from
and hybrid particles. A series of novel nanoparticles has also been developed in the microfluidic
microfluidics, the monotonous structure is gradually changing to complex. More and more works
system. For microfiber materials synthesized from microfluidics, the monotonous structure is
have reported on hybridized microfibers and their functionalization. Meanwhile, the combination
gradually changing to complex. More and more works have reported on hybridized microfibers and
of nanospinning technology and microfluidics enables the nanofiber better performance and novel
their functionalization. Meanwhile, the combination of nanospinning technology and microfluidics
functions. However, there is relatively little work on the microfluidic sheet and construct forms
enables the nanofiber better performance and novel functions. However, there is relatively little
of functional material synthesis, because the increase of material dimension needs complexity
work on the microfluidic sheet and construct forms of functional material synthesis, because the
enhancement of the microfluidic design, which should be improved by developing more operational
increase of material dimension needs complexity enhancement of the microfluidic design, which
should be improved by developing more operational microfluidic technology in the future. In stark
contrast, microfluidic synthesis of nanoparticles and microfibers are in dynamic development. Table
1 summarizes the dimensions of product, chip design class, materials, and biological applications of
biomaterials that are discussed in this review.
Micromachines 2017, 8, 255 20 of 29

microfluidic technology in the future. In stark contrast, microfluidic synthesis of nanoparticles and
microfibers are in dynamic development. Table 1 summarizes the dimensions of product, chip design
class, materials, and biological applications of biomaterials that are discussed in this review.

Table 1. Examples of biomaterials synthesized in microfluidic platform.

Dimension of Potential Biomedical

Chip Design Materials Reference
Product Applications
0D (spherical “Squeezing out” Not mentioned in
Edible oil [37]
microparticles) microchannel original work
Poly tripropyleneglycol
0D (spherical Not mentioned in
Flow-focusing diacrylate (polyTPGDA) [38]
microparticles) original work
Polyurethane
0D (spherical
Coaxial Polyacrylamide (PAM) Glucose monitoring [39]
microparticles)
0D (spherical
T-junction Calcium alginate Cell carrier [40]
microparticles)
0D (spherical Poly(lactide-co-glycolide)
Flow-focusing Drug delivery [41]
microparticles) (PLGA)
0D (spherical Poly(dl-lactic acid) or
Coaxial Drug delivery [42]
microparticles) polycaprolactone (PCL)
0D (spherical
T-junction variant Poly(lactic acid) (PLA) Drug delivery [43]
microparticles)
0D (spherical Sensors
Flow-focusing Silica (SiO2 ) [44]
microparticles) Biomolecule delivery
0D (special-shaped Not mentioned in
Flow-focusing PolyTPGDA [45]
microparticles) original work
0D (special-shaped T-junction Poly(ethylene glycol) diacrylate Drug delivery
[46]
microparticles) Flow-focusing (PEGDA) Biological probes
0D (special-shaped Cell trapping or
Coaxial PLA [47]
microparticles) immobilisation
0D (special-shaped Controlled release
Flow-focusing SiO2 [48]
microparticles) Biosensing
0D (special-shaped Lithography Drug delivery
PEGDA [49]
microparticles) channel Biosensors
0D (special-shaped Lithography
Colloidal Glass SiO2 Biosensor [50]
microparticles) channel
0D (core–shell Lithography
PEGDA Cell assembly [51]
microparticles) channel
0D (core–shell Flow-focusing Ferrofluid Magnetic imaging
[53]
microparticles) Double emulsions PAM Micro-mixing
PEGDA
0D (core–shell Coaxial Bioassays
Ethoxylated trimethylolpropane [54]
microparticles) Double emulsions Cell culture
triacrylate (ETPTA)
0D (core–shell Coaxial Polyethylene glycol (PEG)
Biomolecular sensing [55]
microparticles) Double emulsions Colloidal nanosensors
0D (core–shell
Flow-focusing PEGDA Agent delivery [56]
microparticles)
0D (core–shell ZIF-8
Coaxial Drug carrier [57]
microparticles) Alginate
0D (core–shell
Flow-focusing SiO2 Detoxification [58]
microparticles)
0D (core–shell Ultrasonic
T-junction Poly(styrene-co-acrylic acid) [59]
microparticles) MRI
Micromachines 2017, 8, 255 21 of 29

Table 1. Cont.

Dimension of Potential Biomedical

Chip Design Materials Reference
Product Applications
SiO2
0D (porous
Coaxial Silver Molecular detection [62]
microparticles)
ETPTA
0D (porous Carriers of biologically
Flow-focusing Poly(GMA-co-EGDMA) [63]
microparticles) active species
0D (porous Lithography
PEG Biomolecule analysis [64]
microparticles) channel
0D (porous Lithography
PEG Protein detection [65]
microparticles) channel
0D (porous Drug carrier
Flow-focusing PLGA [66]
microparticles) Cell carrier
0D (porous
Coaxial PLGA Cell scaffold [67]
microparticles)
0D (composite Silver
Flow-focusing Antibacterial [68]
microparticles) Chitosan
Pectin
0D (composite
Flow-focusing Alginate Controlled release [69]
microparticles)
Biopolymer
Coaxial
0D (composite
Multi-Barrelled Calcium alginate Cell carrier [70]
microparticles)
Capillary
0D (composite Lithography PEGDA
Drug release [71]
microparticles) channel Ketal-containing diacrylamide
PAM
0D (composite Magnetically
Coaxial Poly(N-isopropylacrylamide) [72]
microparticles) manipulation
Iron oxide particles
Magnetically
0D(nanoparticles) 3D flow-focusing Goethite [77]
manipulation
SiO2 -coated magnetic
0D(nanoparticles) 3D flow-focusing MRI [78]
nanoparticles
Digital droplet
0D(nanoparticles) Supramolecular nanoparticles Molecular imaging [79]
generator
0D(nanoparticles) Single channel Silver Biosensing [80]
0D(nanoparticles) Single channel Gold Biosensing [81]
Biosensor for protein
0D(nanoparticles) Single channel Gold and polypeptide [82]
detection
0D(nanoparticles) T-junction variant Au/Ag/Au Plasmonic application [83]
0D(nanoparticles) 2D flow-focusing PEG-PLGA Drug delivery [84]
0D(nanoparticles) 3D flow-focusing PEG-PLGA Drug delivery [85]
0D(nanoparticles) 3D flow-focusing Lipid-PLGA Drug delivery [86]
0D(nanoparticles) 3D flow-focusing Lipid-PLGA Drug delivery [87]
PEO45 -b-PS45
0D(nanoparticles) 2D flow-focusing Platinum nanoparticles Drug delivery [88]
Gold nanorods
0D(nanoparticles) T-junction Polyplexes Nucleic acid delivery [89]
Y-junction
0D(nanoparticles) Herringbone Lipid Nucleic acid delivery [90]
structures
0D(nanoparticles) 2D flow-focusing Chitosan Nucleic acid delivery [91]
0D(nanoparticles) 3D flow-focusing Lipid-PLGA Nucleic acid delivery [92]
Micromachines 2017, 8, 255 22 of 29

Table 1. Cont.

Dimension of Potential Biomedical

Chip Design Materials Reference
Product Applications
Digital droplet
0D(nanoparticles) Supramolecular nanoparticles Nucleic acid delivery [93]
generator
Cancer treatment
0D(nanoparticles) Coaxial Lipoplexes [94]
potential
Tetrathiafulvalene Not mentioned in
0D(nanoparticles) 3D flow-focusing [96]
Au original work
0D(nanoparticles) 3D flow-focusing Polyplexes Therapeutics [97]
1D(microfibers) Flow-focusing Calcium alginate Tissue engineering [99]
1D(microfibers) T-junction variant PLGA Tissue engineering [100]
1D(microfibers) Flow-focusing Calcium alginate Tissue engineering [101]
Methacrylamide-modified
1D(microfibers) T-junction Tissue engineering [102]
gelatin or alginate
Cells or biomolecules
1D(microfibers) Flow-focusing Calcium alginate [103]
carrier
1D(microfibers) Flow-focusing Calcium alginate Cells or drug delivery [104]
1D(microfibers) Flow-focusing Calcium alginate Drug carriers [105]
1D(microfibers) Coaxial Chitosan Drug carriers [106]
1D(nanofibers) Flow-focusing Calcium alginate Biomimetic material [108]
1D(nanofibers) Y-junction PLGA Tissue engineering [109]
Multiple parallel
2D(sheet) Calcium alginate Tissue engineering [110]
microchannels
Specifical Drug delivery
2D(sheet) Silica [111]
patterning Cell guidance
Multiple parallel
2D(sheet) Calcium alginate Tissue engineering [112]
microchannels
Drug delivery
2D(sheet) T-junction Alginate [113]
Tissue engineering
Specifical Silk fibroin
3D(constructs) Tissue engineering [115]
patterning Chitosan
Specifical
3D(constructs) Collagen Tissue engineering [116]
patterning
Specifical
3D(constructs) Calcium alginate Tissue engineering [117]
patterning
3D(constructs) Flow-focusing Calcium alginate Tissue engineering [118]
3D(constructs) Flow-focusing Chitosan Tissue engineering [119]
3D(constructs) Coaxial Calcium alginate Tissue engineering [120]
3D(constructs) T-junction Calcium alginate Tissue engineering [121]
3D(constructs) Flow-focusing Calcium alginate Tissue engineering [122]

Based on the development of these microfluidic synthetic technologies, multi-scale structured

biomaterials have displayed remarkable properties, including various chemical compositions,
high specific surface area and multiscale interface structures, which would be widely applied in the
fields of biomedicine, such as drug carrier, cell scaffold, and cellular microenvironment construction.
Although microfluidic synthesis has shown significant advantages in both technical and specific
applications, as a research field which is at a relatively initial development period, it still faces
some pressing problems. Firstly, since current works mostly adopt single-channel as the reaction
unit, the yield of microfluidic synthetic materials is low compared to conventional batch mode.
To solve this bottleneck, the micro-units in the microfluidic system could be repeated to improve
the reaction throughput. Large-scale productions of the microfluidic biomaterials are as shown in
Micromachines 2017, 8, 255 23 of 29

Figure 17 [123–125], which should be guidance for the industrialization of microfluidic synthesis.
The working principles of these three microfluidic modules are based on coaxial annular interfaces
(Figure 17a), stacking multiple generator layers (Figure 17b), and parallel multiple modular reactors
(Figure 17c), respectively. Next, to highlight the integration strength of the microfluidic platform,
automation control should be enhanced to reduce manual operation. Finally, to design and prepare
novel biomaterials with more applications, microfluidic platform advantages and practical application
of the biomaterials should be dynamically combined. In the future, microfluidic biomaterial design
Micromachines 2017, 8,should
and preparation 255 be application targeted and oriented. 23 of 29

100 μm

1 mm 100 μm

500 μm

Figure 17. Scaled-up synthesis of functional biomaterials based on microfluidics. (a) High-volume
Figure 17. Scaled-up synthesis of functional biomaterials based on microfluidics. (a) High-volume
production of emulsions in a microfluidic parallelization arrangement; (b) A liter per hour volume
production of emulsions in a microfluidic parallelization arrangement; (b) A liter per hour volume
production of single emulsions in a parallelization chip; (c) Particle synthesis in parallel multiple
production of single emulsions in a parallelization chip; (c) Particle synthesis in parallel multiple
modular
modular microfluidic
microfluidic reactors.
reactors. Reproduced
Reproduced with
with permission
permission from
from [123–125].
[123–125].

Acknowledgments: This work was funded by the National Natural Science Foundation of China (No.
81271412, 81471308, 31500793), International S&T Cooperation Project of the Ministry of S&T of China (No.
Acknowledgments: This work was funded by the National Natural Science Foundation of China (No. 81271412,
2010DFR30850), the Scientific
81471308, 31500793), Research
International Foundation Project
S&T Cooperation for Returned OverseasofChinese
of the Ministry S&T of Scholars,
China (No.and the Doctoral
2010DFR30850),
Scientific Research Foundation of Liaoning Province (No. 201501158).
the Scientific Research Foundation for Returned Overseas Chinese Scholars, and the Doctoral Scientific Research
Foundation of Liaoning Province (No. 201501158).
Author Contributions: Jingyun Ma performed the document indexing and manuscript writing. Yachen Wang
performed the document
Author Contributions: indexing
Jingyun and figure
Ma performed theprocessing. Jing Liuand
document indexing proposed and writing.
manuscript designed the whole
Yachen Wang
performed
manuscript. the document indexing and figure processing. Jing Liu proposed and designed the whole manuscript.
Conflicts of Interest: The authors declare no conflict of interest.
Conflicts of Interest: The authors declare no conflict of interest.

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