CHAPTER I

THE INTRODUCTION

This chapter states the reason of the researcher for pursuing their topic and the

problems that will be answered at the process of the experiments. The hypotheses, both

null and alternative are also included, together with the rationale of the study. In this

chapter, the scope and the limitations of the study are read including the related studies and

literature that the researcher used as a handy tool in the completion of the study.

Background of the Study

Surgical sutures are staple instruments in the field of medicine, and is by far the

most common. Often utilized to prevent bacterial infection, stop bleeding wounds

instigated by injuries and reduce acute scarring, the technique involving suturing dates back

into Ancient Egypt circa 30,000 years ago. As medical practitioners since then have

continued to advance their work, a wide array of materials have been used to produce

different types of sutures, that of which includes naturally found constituents such as silk,

cotton, linen, and catgut, all of which depending on what specific part of the body the suture

is going to be used. Synthetic materials have also been used to produce commercially

available sutures and is, in fact, still used in modern medicine, one of which is polymer.

(Struszczyk, 2002)

Polymer based sutures are known to satisfy most of the physical (sterility, tensile

strength, uniformity in diameter, and pliability) and chemical properties a medical suture

presents. Although still not ideal, it has gained its immense attraction from scientists and

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health practitioners alike due to its carbon-based chemistry which is considerably closer to

biological tissue than inorganic materials. (Maitz, 2015)

It is in this regard that a type of naturally occurring polymer found in the shells of

the Penaeus monodon, common name Asian Tiger Shrimp, endemic to the Philippine

waters, called chitosan is now being viewed of the researchers of the study as a potential

non-synthetic material to be used for medical sutures. A chitosan-based suture presents a

low-inflammatory response material and is significantly much less painful to remove as

opposed to commercially-made sutures (Montenegro & Godeiro, 2012) as it shows a

biological aptitude to stimulate cell proliferation and tissue organization. It can also play

the role of a biological primer for cell-tissue proliferation and reconstruction. (Muzzarelli

et. al, 1988)

Statement of the Problem

General Problem

This study aims to utilize chitosan fibers from Penaeus monodon (Asian

Tiger Shrimp) shells to create an ideal medical suture and to test the chitosan’s

feasibility as a main constituent in the medical suture.

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 Specific Problems

Specifically, this study attempts to answer the following problems:

1. Will chitin be extracted from Penaeus monodon (Asian Tiger Shrimp)

shells be converted from chitin through deproteinization and

demineralization?

2. Will the chitin be converted into chitosan fibers through the Alkaline

Hydrolysis Method and Wet-Spinning Technique?

3. Will there be a significant difference in the chitosan-derived surgical

sutures and the commercially available sutures in terms of its general

mechanical properties which are as follows:

o Sterility

o Uniformity

o Pliability for ease of handling and knot security

o Uniform tensile strength by suture type and size

o Freedom from irritants or impurities that would elicit tissue

reaction

Hypotheses

The researchers came up with possible hypotheses to the problems that they might

encounter in the study, Chitosan Fibers from Penaeus monodon (Asian Tiger Shrimp)

shells as Medical Suture. Below are the researchers’ hypotheses in null and alternative

form.

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Null

1. Chitosan fibers from Penaeus monodon (Asian Tiger Shrimp) shells cannot

create an ideal medical suture and the chitosan is not feasible as a main

constituent for medical suture.

2. Chitin cannot be extracted from Penaeus monodon (Asian Tiger Shrimp) shells

and cannot be converted from chitin through deproteinization and

demineralization.

3. Chitin cannot be converted into chitosan fibers through the Alkaline Hydrolysis

Method and Wet-Spinning Technique.

4. There is no significant difference in the chitosan-derived surgical sutures and

the commercially available sutures in terms of its general mechanical

properties.

Alternative

1. Chitosan fibers from Penaeus monodon (Asian Tiger Shrimp) shells can create

an ideal medical suture and the chitosan is feasible as a main constituent for

medical suture.

2. Chitin can be extracted from Penaeus monodon (Asian Tiger Shrimp) shells

and can be converted from chitin through deproteinization and

demineralization.

3. Chitin can be converted into chitosan fibers through the Alkaline Hydrolysis

Method and Wet-Spinning Technique.

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 4. There is a significant difference in the chitosan-derived surgical sutures and the

commercially available sutures in terms of its general mechanical properties.

Theoretical Framework

A pivotal theory which underpins this study is George Winter’s (1962) Moist

Wound Healing Theory. While the central concern of this thesis is with regard to Chitosan

effectiveness, the foundations of the study would be nowhere found without its anchorage

on healing theory—a component which serves as one of the objectives sought by the

researchers. In order to better understand surgical methods and their instruments thereof, it

is of great import to refer to propositions which have laid the groundwork to their

perfection, that is, their healing or curing properties.

Chitosan is reputable in its own right, yet to test its viability as an added substance

to the creation of a surgical suture, the fundamentals of wound dressing must be addressed.

The aforementioned premise thus begs the researchers to seek a salient theory therein

which has fostered immense succor to the field of surgical science—the Moist Wound

Healing Theory. According to this theory by Winter (1962 as cited in Bryan, 2004)

epithelialisation occurred twice as fast for wounds kept moist as opposed to dry wounds.

The healing optimization afforded by the moist environment not only aids significantly

towards the betterment of scarred patients, but also to the application of their surgical

dressings thereof. Moreover, Winter also affirms (1962 as cited in Bryan, 2004) that the

incidence of a wound being infected would decrease if the surgical process would be

conducted in this ideal type of atmosphere. Such finding has thus been groundbreaking in

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medical field and practice as it would expand the available literature and knowledge on

wound dressing and healing. This theory has also been backed up by numerous researchers,

most importantly Hinman and Maibach (1963 as cited in Bryan, 2004) who have tested the

theory on humans; as it must be noted that Winter has only examined his theory

preliminarily on pigs. As the researchers of this thesis shall test the effectiveness of the

surgical suture, this theory will be of great assistance having borne in mind and practice,

for the dryness or moistness of the surrounding environment can be a factor in the efficacy

of Chitosan-based surgical sutures.

On another note, Tachibana et al.’s (1988) theory of Chitin suture strength

reinforces a primary objective by the researchers, namely, to prove the effectiveness of the

surgical suture infused with fibers from the said substance. According to them, the

properties possessed by a Chitin-based suture made it a viable choice as a surgical aid and

instrument (Tachibana et al., 1988). For one, its smaller elongation capacity equates to a

lower probability of loosening once applied to wounds. Another advantage it affords is its

tensile strength which spells for a secure and sturdy implement for wound dressing.

Ultimately, Tachibana et al.’s research has led them to conclude that “Chitin is a suture

material which is easily ligated and difficult to loosen” (1988, p. 538), thus fortifying the

bedrock of this thesis.

Conceptual Framework

The researchers will gather their data from Penaeus monodon (Asian Tiger
Shrimp), which is commercially available and can be easily bought from local markets.
The researchers will then proceed onto executing the set of methods that they have deemed
necessary to extract the chitosan from the Penaeus monodon (Asian Tiger Shrimp) shells
namely, (a) manual head and carapace separation via Raw Material Preparation, (b)

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deproteinization and demineralization through Chitin Extraction, (c) Alkaline Hydrolysis
Method, and lastly the (d) Wet Spinning Techniques which will result in the needed
chitosan-based sutures. The data that will be gathered via the methods specified by the
researchers will be used to attempt to answer the specific problems brought forward in the
earlier parts of the study.

OUTPUT

Chitosan fibers from

INPUT
Data from the tests the
researchers of the
study have collected
from the Penaeus
monodon (Asian Tiger
feasibility, properties,
Shrimp) shells
PROCESS
The researchers will
follow the set of pre-
determined methods
in order to come up
with the chitosan-
based suture and
determine the its
and significant
difference through a
series of tests.
Penaeus monodon
(Asian Tiger Shrimp)
shells can create an
ideal medical suture
and is feasible as a
main constituent in
the medical suture.
---------or---------
Chitosan fibers from
Penaeus monodon
(Asian Tiger Shrimp)
shells cannot create an
ideal medical suture
and is not feasible as a
main constituent in
the medical suture.

Significance of the Study

The notable significance of the availability of medical tools in the field of medicine

has been proved countless of times along with the modernization of human civilization.

Progressions in the medical field have provided humanity with better solutions to answer

the most confounding cases with regards to the body through the aid of science and

research. In attempting to create an ideal medical suture out of a naturally occurring

biopolymer which is chitosan, the researchers of the study are not only aiming to find

benefit for themselves but also to the medical community, the government branch which

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deals with the citizens’ health-related concerns (Department of Health), and the country

itself. Additional sufficient knowledge is quite enough for the researches of the study to

properly execute the creation of the expected product. The set of procedures will enable

them to observe the outcome of their research which will enable them to properly reach

their inferences and conclusions before, during, and after the study is conducted.

Patients and medical practitioners alike will benefit from the study as well since

actual testing and usage of the resulting product will be utilized by these people. The

response they will have are going to be crucial in acquiring most of the vital information

the researchers of the study will use. This will include answers to the problems posed in

the earlier part of the study.

Providing cheap alternatives without comprising quality is one of a government

institution’s goal in responsibly taking care of a certain country’s citizens. The Department

of Health (DoH) is one of these government institutions. Feasibility of the expected product

provides them another option when dealing with the distribution of safe and efficient

medical tools to the general masses.

In achieving a goal this significant will also benefit the country in a major scale. It

will then be provided with an innovative tool that will be able to exhibit the properties of

commercially made sutures that will potentially aid in providing better results in terms of

wound healing and recovery.

Scope and Delimitations of the Study

The study’s purpose is to utilize chitosan fibers from Penaeus monodon (Asian Tiger

Shrimp) shells as medical sutures.

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 Chitosan is a natural biopolymer abundant from different sources like crabs and

lobster shells, insects, and fungi. This study, however, will only utilize one source of

chitosan and that is from shrimp. The researchers of the study will also be limiting the

source further into only one specific specie of shrimp, the Penaeus monodon (Asian Tiger

Shrimp) as it is the most common type that can be found in the Philippine setting. The

researchers will also not be able to test the suture on a live organism as time constraints

and possible additional requirements with regards to animal and/or human welfare must be

met and accomplished.

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Definition of Key Terms

 Biopolymer (noun)

o a polymeric substance (such as a protein or polysaccharide) formed in a

biological system

 Carapace (noun)

o a bony or chitinous case or shield covering the back or part of the back of

an animal (such as a turtle or crab)

 Catgut (noun)

o a tough cord made usually from sheep intestines

 Chitin (noun)

o a horny polysaccharide that forms part of the hard outer integument

especially of insects, arachnids, and crustaceans

 Chitosan (noun)

o a substance formed from chitin by partial deacetylation with alkali

 Crustacean (noun)

o any of a large class (Crustacea) of mostly aquatic mandibulate arthropods

that have a chitinous or calcareous and chitinous exoskeleton, a pair of often

much modified appendages on each segment, and two pairs of antennae and

that include the lobsters, shrimp, crabs, wood lice, water fleas, and barnacles

 Litigate (intransitive verb)

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 o to carry on a legal contest by judicial process

 Polymer (noun)

o a chemical compound or mixture of compounds formed by polymerization

and consisting essentially of repeating structural units

 Shrimp (noun)

o any of numerous mostly small and marine decapod crustaceans (suborders

Dendrobranchiata and Pleocyemata) having a slender elongated body, a

compressed abdomen, and a long spiny rostrum and including some

(especially family Penaeidae) that are commercially important as food; also

: a small crustacean (such as an amphipod or a branchiopod) resembling the

true shrimp

 Suture (noun)

o a stitch made with a suture

o a strand or fiber used to sew parts of the living body

o the act or process of sewing with sutures

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 CHAPTER II

REVIEW OF RELATED LITERATURE

The researchers gathered legal bases, literature, and other studies that had

comparable similarities with the study they are conducting. The resources come from both

local and foreign sources. All the information they gathered served as the basis of their

study and aided them in understanding their study in a deeper sense.

Related Readings

A. Foreign

According to Section 878 of the Code of Federal Regulations Title 21, medical

devices are classified into Class I, II and III and regulatory requirements and controls are

defined by these classifications. The regulatory control increases from Class I to III.

Absorbable sutures such as polyglycolide and poly(l-lactide) sutures are under Class II and

these devices require general and special controls. General controls consist of pre-market

notification or the “510(k) Program”, properly labeling of the medical device, and

compliance with the U.S. Food and Drug Administration’s quality system regulations.

Special controls for these devices require guidelines in the application, usage and handling

of the device, special labeling, and a guidance document.

B. Local

According to Section 4 of the Republic Act No. 8503 entitled “Health Research and

Development of 1998”, the National Institute of Health is established to promote science,

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technology research, and development in the field of health. Innovative ideas are

encouraged to enhance current medical techniques, medical equipment and medical drugs

to ensure continuous development. For the reason that it will help attain self-sufficiency

and global competitiveness in the manufacturing of health products and devices.

Related Literature

In 2007, Singh and Ray in their article on the Journal of Macromolecular Science

said that chitosan, derived from the second most abundant organic resource known as

chitin, has advantages in biomedical applications, especially in manufacturing absorbable

sutures, with its properties such as nontoxicity and biodegradability. This polymer is a

biocompatible material that disintegrates into harmless products such as amino sugars,

which can be eventually absorbed completely by the body. Sutures made from chitin, which

is the source of chitosan, is used specially when in contact with bile, urine, and pancreatic

juices, which are problem areas with other sutures. In addition, one of the potentials of

chitosan in biomedical applications is that it has also been claimed to accelerate and aid in

wound-healing process. An investigation on the possibility of chitosan as wound healing

accelerator done by Malette, Quigly, and Adickes show that no abnormal tissue reactions

or infections were observed in using chitin sutures.

Montenegro and Godeiro mentioned in their article entitled “Chitosan Based Suture

– Focusing on the Real Advantages of an Outstanding Biomaterial” that monofilament

fibers are more suitable especially in closing wounds because unlike multifilament sutures,

they do not produce a capillary effect that could transport microorganisms from one wound

to another. For this reason, chitosan has antimicrobial properties and does not elicit a high

inflammatory response as it can be manufactured as a monofilament fiber with required

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mechanical properties. The need for removal of sutures is not a necessity for a

biodegradable suture but their degradation time undefined. In other cases, removal of

suture even with biodegradable ones is needed because complications may occur. With

chitosan sutures, applying an acidic solution for few hours dissolves the suture from the

skin instead of in vitro removal, which is a pain-free method.

Related Studies

In a study conducted by Rivelino Montenegro and Thomas Freier, chitosan has been

suggested as an acceptable candidate for bioabsorbable surgical sutures because of its high

biocompatibility. The chitosan fibers’ biocompatibility was tested by implanting the suture

subcutaneously in the neck of the mice that served as the sample. Then, weeks after, the

suture was removed and the area of implantation was examined histologically. There are

no reported signs of inflammation or other significant changes. The result shows that

chitosan is compatible with the living tissue of the organism. On the other hand, a research

led by Kyung-Hye Jung in 2007 suggests the possible anti-bacterial propertyof

PET/chitosan nanofibrous mats. The polymers of Polyethylene terephthalate (PET) and

chitosan were mixed through the process called electrospinning technique on to the PET

micro-nonwoven mat to be used for biomedical applications. The nanofibers of

PET/chitosan were distributed evenly on its surface with a diameter of 500 and 800 nm.

The antibacterial activity of the PET/chitosan nanofiber mats were tested by evaluating its

inhibitory property towards Staphylococcus aureus and Klebsiella pneumoniae. The result

of the evaluation showed a significantly higher growth inhibition rate of the PET/chitosan

nanofiber mats compared to the PET nanofiber mats. Aside from this, the addition of

chitosan into the mat enhanced the wettability of the PET nanofibers. Lastly, the

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PET/chitosan nanofibers mats shows better tissue compatibility than PET nanofibers mats.

Lastly, a study led by Mi-Sun Kim in 2002 shed light regarding the inhibitory and anti-

inflammatory attributes of water-soluble chitosan. A chronic inflammatory response

between β-amyloid (Aβ) and interleukin- 1β (IL-1β) is responsible for the pathology of

Alzheimer’s disease. The cytotoxicity was evaluated to test the biological effect of water-

soluble chitosan production of pro-inflammatory cytokine and inducible nitric-oxide

synthase (iNOS) in human astrocytoma cells galvanize with Aβ and IL-1β particles. The

expression of iNOS stimulated with Aβ and IL-1β and was partially regulated by the usage

of water-soluble chitosan. The result shows the regulatory effects of water-soluble chitosan

to human astrocytoma cells. In line with this, anti-inflammatory property of water-soluble

chitosan was exhibited and this research proved that water-soluble chitosan may reduce

and delay Alzheimer’s disease pathological events.

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