Introduction of Crosslinked Chitosan and Beaded Alginate to Bone Cement

Hannah Kollar
Approximately 1 million procedures are performed each year to repair bone defects,
making bone the second most transplanted tissue [1]. A commonly used bone graft material used
is calcium phosphate cement (CPC) due to its abilities to self harden, easily mold, fully degrade,
and its osteoconductive properties[4]. CPC is used for applications such as fracture augmentation
and screw and metal implant fixation [1]. CPCs are generally made using a mixture of
tetracalcium phosphate and dicalcium phosphate anhydrous, however more recently additional
organic and polymeric additives are included to improve varying properties[2]. A major issue with
CPC is its relatively low strength and brittleness, which allows it to remain in only non-load
bearing bone[4]. Porosity is also an important property of CPCs because it assists with bone
ingrowth, however increased porosity also lowers the mechanical properties critical to the
success of the cement[2]. It has been reported that it would be desirable for the CPC implant to
consist of macropores to create a more rapidly resorbing and osseointegrated implant while
keeping the strength and brittleness at appropriate levels[5].
Chitosan is a biologically renewable, biodegradable, biocompatible, nonantigenic, nontoxic, and biofunctional natural cationic polymer[6]. Chitosan is a derivative of chitin a natural
mucopolysaccharide known to be the supporting material of crustaceans and insects[7]. It is
known for promoting cell adhesion, proliferation, and differentiation, antibacterial activity, and
minimizing inflammatory responses while evoking minimal foreign body reaction making it
ideal for tissue engineering[6][8]. According to studies, incorporating chitosan into CPCs show an
increase in injectability and strength with little effect on handling and setting properties[8]. The
introduction of chitosan mesh in bone cements proved in increase strength and flexibility[3].
Once the chitosan begins to degrade a network of interconnected macropores became present in

implant, further promoting bone ingrowth[3]. The creation of interconnected macropores within
the bone cement implant is ideal, however using absorbable meshes to do so is undesired. Using
meshes remove the ability of the CPC to be injectable, which is a critical property. Injectability
allows for any size or shape void to be filled, shortened surgical time, minimally invasive, and
faster overall healing [11]. Due to this, it is believed that crosslinking chitosan using a
photoinitiator post-injection would be ideal. One study showed the ability to fabricate a
photocrosslinkable water-soluble chitosan using 2-amino ethyl methacrylate (AEMA) to
introduce functional methacrylate groups[9]. This creates a solution that can crosslink upon
exposure of UV rays within 3 minutes[9]. Through this crosslinking, degradation rate and
mechanical stiffness can be controlled in order to optimize for bone cement[9]. It is important for
crosslinked chitosan to degrade slow enough to allow for initial bone ingrowth to integrate into
the implant before creating more porous cement. The strength of the implant gained from the
bony ingrowth and deposition of new bone should offset the weakening of the implant caused by
the increase in porosity[2].
Alginate is an anionic polymer typically obtained from brown seaweed, with high
biocompatibility, low toxicity, and low cost[12]. The addition of alginate hydrogel beads to the
CPC paste will allow for initial porosity. It is reported that most CPC mixtures are fully reacted
and hardened within 1 day, thus it is desirable for alginate beads to degrade quickly after[3]. It
has been shown that the degradation rate of alginate hydrogels can be controlled by partial
periodate oxidation and gamma irradiation, yielding a degradation rate between days to weeks[3].
Pore sizes of 100 600m were shown to promote cell infiltration and bone ingrowth and hence
would be the targeted range for bead size [3].

The final CPC containing both chitosan with AEMA and alginate beads will create a
desirable bone cement. Its improvement in porosity and strength in comparison to CPCs on the
market and ability to remain injectable unlike mesh-enforced CPCs in literature will allow for
further applications in CPCs. More studies must be done in order to define exact ratios of CPCchitosan-alginate to find ideal properties. At the powder-liquid ratio of 3:1, adding uncrosslinked
chitosan to the CPC increased its flexural strength three times and it can be assumed that cross
linking would improve this strength additionally[11]. The addition of chitosan also favorably
decreased the
setting time from
approximately 60
minutes to 6.7
minutes[11]. This
decrease in time
allows for a faster
surgery and less
issues with wash out. The photo-crosslinking process requires 3 minutes of UV light, which can
be completed while the cement implant is fully hardening allowing the procedure to remain
shortened. The elastic modulus and flexural strength readily decreased with an increase in
percent porosity, proving the slow introduction of pores to be favorable as bone ingrowth
progresses. Overall, the cement should display properties similar to that of the bone it is
mimicking. For example cortical bone displays an elastic modulus of 6-20 GPa, yield strength of
80-85 MPa, tensile modulus of 5-20 GPa, and a flexural strength of 150-200 Mpa[12]. It is
important to get as close to these ranges as possible while still being fully injectable.

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