Vol 2, No 1 (2021) 24–32

Tilapia fish col­la­gen: Poten­tial as halal bio­ma­te­r­i­al in tis­sue engi­neer­ing applications

Norhi­dayu Muhamad Zain and Moham­mad Naqib Hamdan

Acad­e­my of Islam­ic Civ­i­liza­tion, Fac­ul­ty of Social Sci­ence and Human­i­ties, Uni­ver­si­ti Teknolo­gi Malaysia, 81310 Johor Bahru, Malaysia.

Cor­re­spon­dence should be addressed to Norhi­dayu Muhamad Zain: norhidayu_​mz@​utm.​my

Cite this: Nusan­tara Halal J. 2021, Vol. 2 No.1 pp. 24–32 (Arti­cle) | Received 15 March 2021 | Revised 20 May 2021 | Accept­ed 22 June 2021 | Pub­lished 30 June 2021 | http://​dx​.doi​.org/​1​0​.​1​7​9​7​7​/​u​m​0​6​0​.​2​0​2​1​v​2​p​0​2​4​-​032

Abstract

Col­la­gen is a nat­ur­al bioac­tive poly­mer wide­ly uti­lized in tis­sue engi­neer­ing appli­ca­tions due to its bio­com­pat­i­bil­i­ty and biodegrad­abil­i­ty. Col­la­gen derived from mam­malian sources such as porcine and bovine is com­mon­ly used as bio­ma­te­ri­als. How­ev­er, due to reli­gious con­cerns, the halal sta­tus of col­la­gen must be put into con­sid­er­a­tion. Since most of the mam­malian col­la­gen is ham­pered by its haram ori­gins, marine col­la­gens are wide­ly inves­ti­gat­ed as alter­na­tives for mam­malian col­la­gen in tis­sue engi­neer­ing appli­ca­tions. Even though the marine col­la­gens are safe and easy to extract, these sources of col­la­gen are hin­dered by their low dena­tur­ing tem­per­a­ture. Tilapia fish (Ore­ochromis niloti­cus) has long been stud­ied for its poten­tial to sub­sti­tute mam­malian col­la­gen for bio­med­ical pur­pos­es due to its high­er ther­mal sta­bil­i­ty com­pared to oth­er marine sources. We here­in review the poten­cy of tilapia col­la­gen as a bio­ma­te­r­i­al for tis­sue engi­neer­ing appli­ca­tions. In this review paper, we main­ly focus on the appli­ca­tion of tilapia col­la­gen in the skin, bone/​dentin, neur­al and corneal tis­sue engineering.

Key­words: Tilapia fish col­la­gen, tis­sue engi­neer­ing, halal bio­ma­te­r­i­al, biomedical.

Introduction

In Islam, halal is defined as per­mis­si­ble (not for­bid­den by Shari­ah), where­as haram is defined as not per­mis­si­ble. Halal is no longer con­sid­ered a mere Muslim’s respon­si­bil­i­ty, but it is con­sid­ered a way of life for Mus­lims as well as non-Mus­lim world­wide. The halal sta­tus is not lim­it­ed to only dietary intake, but it cov­ers a lot of oth­er areas such as cos­met­ic, phar­ma­ceu­ti­cal, and med­ical device prod­ucts.  In 2019, the Depart­ment of Islam­ic Advance­ment of Malaysia (JAKIM) has expand­ed halal cer­ti­fi­ca­tion for med­ical devices, and the stan­dard was pub­lished as “MS 2636 2019 Halal Med­ical Device-Gen­er­al Require­ment” by Stan­dards Malaysia due to demand in pro­pos­als for spe­cif­ic new prod­ucts, chang­ing in the clas­si­fi­ca­tion of med­ical prod­ucts and the use of crit­i­cal ingre­di­ents in man­u­fac­tur­ing med­ical devices.

Col­la­gen is known as a crit­i­cal ingre­di­ent in the devel­op­ment of var­i­ous med­ical devices includ­ing devices derived from tis­sue engi­neer­ing (TE) tech­nol­o­gy due to its nature of ori­gins. Tra­di­tion­al­ly, most of the col­la­gen is extract­ed from mam­malian sources such as porcine and bovine. How­ev­er, these sources are ham­pered by lim­it­ed appli­ca­tions due to reli­gious con­cerns (halal sta­tus) and the risk of dis­eases such as bovine spongi­form encephalopa­thy and aph­t­hous fever dis­ease [1]. There­fore, a vari­ety of marine sources were iden­ti­fied as a safe source of col­la­gen and now replac­ing mam­malian col­la­gen for TE pur­pos­es. More­over, the extrac­tion of col­la­gen from the fish by-prod­ucts would lessen the envi­ron­men­tal impact cre­at­ed dur­ing the decom­po­si­tion process and gives an added val­ue to these wastes. How­ev­er, unlike col­la­gen from ter­res­tri­al sources, that of aquat­ic ori­gin have few dis­ad­van­tages in terms of sources depen­dent com­po­si­tion vari­a­tion and low denat­u­ra­tion tem­per­a­ture [2]. Thus, improve­ment of physic­o­chem­i­cal and bio­log­i­cal prop­er­ties of marine col­la­gen are required so that they can be effec­tive­ly employed as scaf­folds for bio­med­ical applications.

Tilapia fish (Ore­ochromis niloti­cus) is one of the main fish groups used to devel­op bio­ma­te­ri­als for TE due to its high­er denat­u­ra­tion tem­per­a­ture than oth­er marine groups. Over the years, more research has proved the poten­tial of the col­la­gen extract­ed from skins, scales, and bones of tilapia in sub­sti­tut­ing mam­malian col­la­gen for TE pur­pos­es. We here­in review the poten­cy of tilapia col­la­gen as a bio­ma­te­r­i­al in TE applications.

Tilapia fish collagen

The tilapia fish species are native to Africa and Mid­dle East and now has been cul­ti­vat­ed and cross­breed in almost all trop­i­cal cli­mate coun­tries and sub­trop­i­cal regions. Tilapia species has become the most impor­tant food fish and was known as aquat­ic chick­en due to their fast growth rate, adapt­abil­i­ty to a wide range of envi­ron­ment con­di­tions, high rate of repro­ducibil­i­ty, easy feed and pro­cess­ing [3]. Ore­ochromis is the most com­mon genus being cul­ti­vat­ed and cross­breed pro­duc­ing hybrids. Nile tilapia (Ore­ochromis niloti­cus), blue tilapia (Ore­ochromis aureus), and tilapia from Mozam­bique (Ore­ochromis mossam­bi­cus) are the com­mon tilapia species being cul­ti­vat­ed in most of the coun­tries. Dur­ing indus­tri­al pro­cess­ing, approx­i­mate­ly 60%-70% of by-prod­ucts are being pro­duced, includ­ing skin, scales, and bones [4]. These parts are rich in col­la­gen and oth­er bioac­tive mol­e­cules. Most of the time, col­la­gens are extract­ed from Nile tilapia (Ore­ochromis niloti­cus) species.

Tilapia fish collagen – applications in TE

Tis­sue injury or organ fail­ure due to severe dis­ease or trau­ma becomes a major health­care prob­lem. The avail­able options such as tis­sue or organ trans­plan­ta­tion are lim­it­ed by the acces­si­bil­i­ty of a com­pat­i­ble donor and could be very cost­ly  [5]. There­fore, TE, an inte­gra­tion of bio­log­i­cal sci­ence and engi­neer­ing to regen­er­ate bio­log­i­cal sub­sti­tute for repair­ing or replac­ing a dam­aged tis­sue or organ, gives a bet­ter alter­na­tive. TE involves three com­po­nents; cells, scaf­fold (3D poly­mer­ic matrix), and growth fac­tors [6]. Among these three com­po­nents, scaf­fold acts as an impor­tant medi­um for restor­ing, main­tain­ing, and improv­ing tis­sue func­tion [7].  Scaf­fold plays its role in tis­sue repair and regen­er­a­tion by pro­vid­ing an appro­pri­ate plat­form, allow­ing the essen­tial sup­ply of numer­ous fac­tors relat­ed to sur­vival, pro­lif­er­a­tion, and dif­fer­en­ti­a­tion of cell [8,9]. Thus, the scaf­fold requires par­tic­u­lar char­ac­ter­is­tics such as it must be bio­com­pat­i­ble and biodegrad­able, pos­sess­es mechan­i­cal prop­er­ties com­pa­ra­ble to the replaced tis­sue, and sup­port cell attach­ment and growth [10]. Most of all, it should mim­ic the ECM in terms of the mor­pho­log­i­cal struc­ture and chem­i­cal com­po­si­tion for the cell attach­ment, pro­lif­er­a­tion, and dif­fer­en­ti­a­tion to be occurred [11]. The selec­tion of bio­ma­te­ri­als to con­struct 3D scaf­fold must be care­ful­ly car­ried out as it has direct influ­ences on cel­lu­lar behaviors. 

Skin tissue engineering

Extreme loss of skin func­tion and struc­ture due to injury and dis­ease may lead to phys­i­o­log­i­cal dis­tur­bances and sub­se­quent­ly major dis­abil­i­ty or even death. Cur­rent advances in TE cat­alyze the devel­op­ment of improved cul­tured skin tis­sue sub­sti­tutes. Tis­sue-engi­neered skin sub­sti­tutes for wound heal­ing have pro­gressed enor­mous­ly over the last cou­ple of years. There are sev­er­al skin scaf­fold types such as porous, fibrous, hydro­gel, micros­phere, com­pos­ite and acel­lu­lar [12]. Syn­thet­ic and high­ly bio­com­pat­i­ble nat­ur­al mate­ri­als have been used to devel­op skin sub­sti­tutes and become alter­na­tives to tra­di­tion­al wound-heal­ing strate­gies and tis­sue regeneration.

In the field of skin TE, col­la­gen-based skin sub­sti­tutes are effec­tive in accel­er­at­ing wound heal­ing by sup­port­ing a suit­able envi­ron­ment for fibrob­last and ker­atinocyte pro­lif­er­a­tion [13], [14]. The most com­mon­ly uti­lized forms of col­la­gen-based bio­ma­te­ri­als for wound heal­ing and TE pur­pos­es are the fib­ril-form­ing col­la­gen [15]. In this fib­ril­lar col­la­gen, fib­rils are formed from the assem­bly of tropocol­la­gen triple helices, which then agglom­er­ate to form fibers [15]. Numer­ous stud­ies have report­ed the use of tilapia fish col­la­gen in dif­fer­ent types of col­la­gen-based scaf­fold for­mu­la­tions for wound and burn to repair, includ­ing col­la­gen-based sponges, elec­tro­spun col­la­gen nanofi­brous, col­la­gen com­pos­ite film, and drug-loaded col­la­gen hydro­gel. The pres­ence of tilapia col­la­gen in the com­pos­ite scaf­folds enhanced sev­er­al prop­er­ties of the skin scaf­folds. For instance,  com­pos­ite porous scaf­folds made of chi­tosan, tilapia fish skin col­la­gen, and glyc­er­ine were proved to facil­i­tate fibrob­lasts and ker­atinocytes infil­tra­tion, adhe­sion, pro­lif­er­a­tion, and sup­port new tis­sue devel­op­ment [16]. In addi­tion, the high amount of fish col­la­gen and glyc­er­ine improved the poros­i­ty, mechan­i­cal strength, biosta­bil­i­ty and cyto­com­pat­i­bil­i­ty of the scaf­folds [16].

In a sim­i­lar study, bet­ter prop­er­ties of chi­tosan-col­la­gen (derived from tilapia fish skin) porous scaf­folds were obtained by incor­po­rat­ing zinc oxide nanopar­ti­cles [17]. In this study, the 2.0% zinc oxide chi­tosan-col­la­gen porous scaf­folds were shown to have the high­est fibrob­last pro­lif­er­a­tion [17]. In a dif­fer­ent study, elec­tro spun PCL/​ col­la­gen (tilapia fish skin col­la­gen) com­pos­ite scaf­folds with dif­fer­ent con­tents of Nile tilapia skin col­la­gen were fab­ri­cat­ed and inves­ti­gat­ed for their bio­log­i­cal activ­i­ties [18]. The results indi­cat­ed that L929 mouse fibrob­lasts were active­ly grown dur­ing the 5 days of cell cul­ture with­out expe­ri­enc­ing cyto­tox­ic effects [18]. Due to the syn­er­getic effects of PCL and col­la­gen, the pro­lif­er­a­tion of L929 fibrob­lasts were found to be sig­nif­i­cant­ly high­er on the PCL/​collagen scaf­fold com­pared to that of con­trol group [18]. The scaf­folds with a col­la­gen con­cen­tra­tion of 8% and 10% were proved to be supe­ri­or to oth­ers in cell adhe­sion and bio­com­pat­i­bil­i­ty [18]. The fish skin col­la­gen might affect intra­cel­lu­lar sig­nal­ing and cell response. In a dif­fer­ent study, the hydrophilic­i­ty of Poly(3‑hydroxybutyrate-co-4-hydrox­y­bu­tyrate) (P(3HB-co-4HB)) films was sig­nif­i­cant­ly increased by incor­po­rat­ing tilapia fish col­la­gen [19]. Sub­se­quent­ly, the col­la­gen blend scaf­fold sur­faces were found to have high­er fibrob­lasts adhe­sion and growth than that of the con­trol. In addi­tion, improved cyto­com­pat­i­bil­i­ty was report­ed in the col­la­gen blend film [19].  Incor­po­rat­ing col­la­gen with oth­er poly­mer mate­ri­als may result in bet­ter prop­er­ties for skin tis­sue sub­sti­tutes. The RGD pep­tide sequence found in col­la­gen is rec­og­nized by the cell sur­face, allow­ing the attach­ment of cells to ECM. 

Bone/​dentin tissue engineering

Crit­i­cal-sized defects in bone main­ly caused by trau­mat­ic injury, bone-relat­ed dis­eases, pri­ma­ry tumor resec­tion, or ortho­pe­dic surgery have in many cas­es may not be capa­ble of repair­ing them­selves by means of mechan­i­cal fix­a­tion alone. These defect sce­nar­ios need a sub­sti­tu­tion­ary mate­r­i­al to fill the bone defect. Sev­er­al bone TE strate­gies, includ­ing acel­lu­lar scaf­folds, gene ther­a­py, growth fac­tor deliv­ery, cell trans­plan­ta­tion, and stem cell ther­a­py, have been applied to address the above issues. Prac­ti­cal­ly, bone TE requires the com­bi­na­tion of the list­ed strate­gies. A tis­sue-engi­neered scaf­fold that mim­ics the com­pli­cat­ed phys­io­chem­i­cal attrib­ut­es of bone may serve as a plat­form to incite the body’s nat­ur­al bio­log­i­cal response to tis­sue dam­age and pro­mote a nat­ur­al heal­ing process that does not occur in crit­i­cal-sized defects. Var­i­ous bio­ma­te­ri­als, includ­ing ceram­ics, met­als, poly­mers, and com­pos­ites, have been stud­ied for their poten­tial as bone scaf­fold mate­ri­als. Nat­ur­al poly­mers espe­cial­ly col­la­gen has been wide­ly uti­lized in bone scaf­fold devel­op­ment due to its bio­log­i­cal fea­tures, net­work and porous struc­tures and mechano-elas­tic behav­ior suit­able for bone TE pur­pose [20]. In addi­tion, col­la­gen fibers become the prin­ci­pal sources of ten­sile strength of bone tis­sues by pro­vid­ing a frame­work for hydrox­ya­p­atite depo­si­tion for fur­ther remod­el­ing [21]. Col­la­gen can be com­bined with oth­er bio­ma­te­ri­als such as hydrox­ya­p­atite, chi­tosan, cal­ci­um phos­phate, and algi­nate to form scaf­folds with dif­fer­ent mechan­i­cal and bio­log­i­cal prop­er­ties [22]. Fish col­la­gen becomes an emerg­ing play­er for bio­med­ical appli­ca­tions due to the patho­log­i­cal risk of mam­malian col­la­gen [23]. Fur­ther­more, fish col­la­gen pep­tides were proved to pro­mote post­tran­scrip­tion­al mod­i­fi­ca­tion for col­la­gen mat­u­ra­tion and gene expres­sion for osteoblasts dif­fer­en­ti­a­tion [24,25].

Unlike skin TE that uti­lized tilapia col­la­gen most­ly from the skin part, col­la­gen derived from tilapia scales is com­mon­ly employed in bone TE appli­ca­tions. For instance, 3D porous scaf­folds were fab­ri­cat­ed by a com­bi­na­tion of tilapia scale col­la­gen and micro­bial trans­g­lu­t­a­m­i­nase (mTGase) enzyme to manip­u­late human mes­enchy­mal stem cells to form osteogenic cells [26]. In this study, mTGase act­ed as a cat­a­lyst to pre­serve the inher­ent prop­er­ties of col­la­gen. The study was con­duct­ed by com­par­ing the per­for­mance of tilapia scale col­la­gen and porcine col­la­gen on the bio­log­i­cal prop­er­ties of the fab­ri­cat­ed scaf­folds. The ALP activ­i­ty of tilapia scale col­la­gen-coat­ed dish and scaf­folds with or with­out mTGase were sig­nif­i­cant­ly high­er than that of porcine col­la­gen sam­ples [26]. These results indi­cat­ed that osteoblas­tic dif­fer­en­ti­a­tion was great­ly enhanced in the pres­ence of tilapia scale col­la­gen with/​without mTGase. Fur­ther­more, the late osteoblas­tic dif­fer­en­ti­a­tion stage of hMSCs was shown 30-fold high­er in the mTGase crosslinked tilapia scale col­la­gen scaf­folds than in the mTGase crosslinked porcine col­la­gen scaf­folds after being cul­tured for 3 weeks [26]. The ear­ly stage of osteoblas­tic dif­fer­en­ti­a­tion in hMSCs was remark­ably accel­er­at­ed on a tilapia col­la­gen sur­face due to spe­cif­ic fib­ril for­ma­tion of tilapia col­la­gen [27]. A fibrous col­la­gen mem­brane was shown to have high­er ALP activ­i­ty than a non-fibrous col­la­gen mem­brane even before adding osteoblas­tic dif­fer­en­ti­a­tion medi­um, sug­gest­ing that the degree of the fib­ril for­ma­tion of tilapia col­la­gen affect­ed the osteoblas­tic dif­fer­en­ti­a­tion of hMSCs. In addi­tion, cal­ci­um depo­si­tion increased sig­nif­i­cant­ly in hMSCs cul­tured on tilapia col­la­gen-coat­ed dish­es com­pared with porcine col­la­gen-coat­ed dish­es, indi­cat­ing tilapia col­la­gen could facil­i­tate the depo­si­tion process [27]. Effect of type I col­la­gen derived from tilapia fish scale on odon­to­blast-like cells was also inves­ti­gat­ed [28]. Bio­com­pat­i­bil­i­ty study of the col­la­gen showed two-fold enhance­ment of the attached cells as com­pared to con­trol. The cells were great­ly induced to dif­fer­en­ti­ate toward odon­to­blast lin­eage as proved by increased ALP activ­i­ty on day 7, improve­ment of ALP, BSP mRNA expres­sion on day 7 and 10, as well as enhanced min­er­al­iza­tion on day 9 [28]. Bio­com­pat­i­bil­i­ty of tilapia scale col­la­gen was also eval­u­at­ed for tis­sue regen­er­a­tion in the oral-max­illo­fa­cial area [29]. Odon­to­blast pro­lif­er­a­tion, dif­fer­en­ti­a­tion, and min­er­al­iza­tion in tilapia scale col­la­gen exhib­it­ed com­pa­ra­ble per­for­mance to porcine col­la­gen [29]. Since the future use of mam­malian col­la­gen may be ham­pered by reli­gious restric­tion, bovine spongi­form encephalopa­thy (BSE), foot and mouth dis­ease, under­uti­lized tilapia scale col­la­gen offers a poten­tial alter­na­tive for the mam­malian col­la­gen and might be use­ful for bone and dentin-pulp regeneration.

Neural tissue engineering

The ner­vous sys­tem is the most impor­tant sys­tem in the body since the sen­so­ry and motor func­tions are high­ly depen­dent on this sys­tem. Injuries to this sys­tem affect the body’s func­tions and could be lethal for humans. How­ev­er, due to the com­plex­i­ty of this sys­tem and its restrict­ed abil­i­ty to regen­er­ate, the restor­ing process has always been a chal­lenge for neu­ro­bi­ol­o­gists and neu­rol­o­gists. To date, sev­er­al sci­en­tif­ic approach­es have been sug­gest­ed to restore the func­tion of a dam­aged ner­vous sys­tem, includ­ing cell ther­a­pies and TE [30]. Nov­el strate­gies that com­bined bio­ma­te­ri­als, cells, and growth fac­tors pro­vide a poten­tial solu­tion to tack­le these neu­ro­log­i­cal dis­or­ders. Induced pluripo­tent stem cells (iPSCs)  hold great poten­tial for cell ther­a­pies and TE [31]. The abil­i­ty of iPSC to dif­fer­en­ti­ate and devel­op into func­tion­al cells is one of the cru­cial com­po­nent in devel­op­ing regen­er­a­tive med­i­cines [31]. A com­bi­na­tion of bio­chem­i­cal fac­tors and mechan­i­cal prop­er­ties of the ECM could deter­mine the fate of stem cells. Tis­sue stiff­ness of the ECM is one of the mechan­i­cal prop­er­ties that affect the deter­mi­na­tion of iPSCs fate toward spe­cif­ic cel­lu­lar sub­types [32]. For iPSCs to dif­fer­en­ti­ate into neur­al lin­eage choice, the stiff­ness con­di­tion of liv­ing brain tis­sue must be repro­duced in vit­ro. A study by Iwashita et al. has suc­cess­ful­ly mim­ic­ked the stiff­ness of liv­ing brain tis­sue in vit­ro using tilapia skin col­la­gen gels [32]. The tilapia col­la­gen gels were crosslinked with a com­bi­na­tion of 1‑ethyl-3-(3‑dimethylaminopropyl)-carbodiimide (EDC) and N‑hydroxy suc­cin­imide (NHS) to pro­duce a steady soft­er range of col­la­gen gel that mim­ics brain tis­sue. A stiff­ness of 150‑1500 Pa (like adult brain stiff­ness) with high repro­ducibil­i­ty was obtained at a ratio of NHS to EDC is 0.1. Pluripo­tent cells were shown to dif­fer­en­ti­ate into neur­al lin­eage and pro­mot­ed the pro­duc­tion of dor­sal cor­ti­cal neu­rons when exposed to the tilapia col­la­gen gels [32]. These find­ings demon­strate that the tilapia col­la­gen gel could be used for neur­al induc­tion from pluripo­tent cells and pro­vide a cru­cial devel­op­ment for neur­al regen­er­a­tive applications.

Corneal tissue engineering

Corneal dam­age is a major cause of blind­ness world­wide, sec­ond only to cataracts. Tra­choma, corneal opac­i­ties, and child­hood blind­ness could lead to corneal blind­ness [33]. Cur­rent­ly, corneal trans­plan­ta­tion is con­sid­ered the main method for visu­al restora­tion treat­ment in corneal blind­ness patients. Full-thick­ness replace­ment of pen­e­trat­ing ker­ato­plas­ty was the first method used to per­form corneal trans­plan­ta­tion and pre­vails as the most com­mon method [34]. Nev­er­the­less, the avail­abil­i­ty of corneal donor tis­sue becomes the fun­da­men­tal prob­lem with the corneal replace­ment method. A severe short­age of donor tis­sue, lim­it­ed access to drugs such as steroid and antibi­ot­ic, and lack of skilled sur­geons, result­ing in increased num­ber of untreat­ed patients [33]. There­fore, arti­fi­cial corneal sub­sti­tutes have been stud­ied to over­come the short­age and prob­lems asso­ci­at­ed with human donor corneas.

Syn­thet­ic pros­the­ses and tis­sue-engi­neered con­structs were devel­oped to facil­i­tate the regen­er­a­tion of the host tis­sue and restore the cornea’s refrac­tive func­tion [35–37]. 3D scaf­folds made from bio­ma­te­ri­als could mim­ic the corneal stro­ma and pro­vide a suit­able envi­ron­ment for the patient’s own corneal cells to repop­u­late and regen­er­ate [35,38,39]. The scaf­folds can be syn­thet­i­cal­ly fab­ri­cat­ed or har­vest­ed in an almost ready state. The human corneal stro­ma com­pris­es main­ly of type I col­la­gen, orga­nized in orthog­o­nal lamel­lae, result­ing in enhanced ten­sile strength in the cornea [40]. Thus, col­la­gen could be the most suit­able mate­r­i­al used to con­struct an arti­fi­cial corneal scaf­fold. In addi­tion, the already exist­ing col­la­gen scaf­fold in nature may reduce the fab­ri­ca­tion cost and promise an ade­quate resource for clin­i­cal trans­plan­ta­tion [41]. Fish scales are pri­mar­i­ly com­posed of con­nec­tive tis­sue pro­tein and col­la­gen (up to 81%), cov­ered with cal­ci­um phos­phate and cal­ci­um car­bon­ate. There­fore, acel­lu­lar and decal­ci­fied fish scales may serve as an effec­tive col­la­gen scaf­fold that induce regen­er­a­tion of the dam­aged cornea by emu­lat­ing the func­tions of the high­ly nat­ur­al ECM scaf­fold­ing of the cornea. 

Tilapia fish scales are known to have par­al­lel-arranged col­la­gen fibers, mim­ic­k­ing the human corneal stro­ma [42]. Sev­er­al pre­vi­ous stud­ies have report­ed the poten­tial of decal­ci­fied tilapia fish scale col­la­gen scaf­folds for corneal regen­er­a­tion [34,41–45]. Lin et al. devel­oped decal­ci­fied tilapia scale col­la­gen scaf­folds to serve as an in vit­ro tem­plate for cul­tur­ing the corneal cells [42]. The nat­ur­al 3D microstruc­ture sus­tains its ini­tial struc­ture even after being acel­lu­lar­ized and decal­ci­fied. The fab­ri­cat­ed scaf­folds dis­played good rab­bit corneal cell pro­lif­er­a­tion and biosyn­thet­ic activ­i­ty after 7 days of cul­ti­va­tion [42]. The micropat­terned struc­tures of the decal­ci­fied scales are not only facil­i­tat­ing cell attach­ment but also guid­ing cell migra­tion through mul­ti­ple par­al­lel chan­nels [42]. In a dif­fer­ent study, the light-scat­ter and light-trans­mis­sion prop­er­ties of tilapia fish scale col­la­gen matrix were inves­ti­gat­ed. The amount of scat­tered light was sim­i­lar to that seen in an ear­ly cataract. Mean­while, the light trans­mis­sion was com­pa­ra­ble to the trans­mis­sion through the human cornea [41]. Rat ker­ato­plas­ty mod­el was used for corneal trans­plan­ta­tion stud­ies at three dif­fer­ent sur­gi­cal sites (ante­ri­or lamel­lar ker­ato­plas­ty-ALK, inter­lamel­lar corneal pock­et-IL, and sub­con­junc­ti­val-SC). Dif­fer­ent degrees of hazi­ness, pupil obscure­ness, and inflam­ma­tion were gen­er­al­ly seen at those implant­ed sites [41]. This exper­i­ment showed that the fab­ri­cat­ed decel­lu­lar­ized scaf­fold has suf­fi­cient light trans­mis­sion val­ues and is suit­able for use in ker­ato­plas­ty.  The same research group per­formed an in-depth study to deter­mine the suit­abil­i­ty of a tilapia fish scale-derived col­la­gen matrix for corneal recon­struc­tion [45]. The results showed no cyto­tox­i­c­i­ty effects, nor­mal phe­no­type mark­ers, and no inflam­ma­tion or sen­si­ti­za­tion. More­over, the implant­ed corneal led to a trans­par­ent cornea, healthy epithe­li­um, and no immuno­genic response [45].

In a sep­a­rate exper­i­ment, mor­pho­log­i­cal and phys­i­o­log­i­cal prop­er­ties of decal­ci­fied tilapia scale col­la­gen implants were stud­ied using 6 months of fol­low-up of rab­bit mod­el [34]. The implant­ed cornea dis­played a clear sur­face with no haze and ulcer detect­ed up to 6 months post­op­er­a­tive­ly. In addi­tion, no immune response, dis­so­lu­tion, frag­men­ta­tion, and degen­er­a­tion were observed after a long-term eval­u­a­tion [34]. The poten­tial of acel­lu­lar and decal­ci­fied tilapia fish scale col­la­gen as an ide­al arti­fi­cial cornea sub­sti­tute was also proved by inves­ti­gat­ing its bio­com­pat­i­bil­i­ty towards pri­ma­ry human corneal endothe­lial cells (HCEnCs) [44]. In line with the pre­vi­ous research, the scaf­folds dis­played cor­rect mor­phol­o­gy, cyto­com­pat­i­bil­i­ty, and no tox­i­c­i­ty for HCEnCs [44]. Pre­vi­ous stud­ies revealed that the new approach of using acel­lu­lar and decal­ci­fied tilapia fish scale col­la­gen scaf­fold might yield an ide­al arti­fi­cial cornea sub­sti­tute for long-term inlay place­ment. How­ev­er, reg­u­la­to­ry com­pli­ances like that of advanced ther­a­py med­i­c­i­nal prod­ucts are need­ed for fur­ther clin­i­cal use.

Conclusions

The review clear­ly nar­rates that tilapia fish col­la­gen has poten­tial uses in TE. Con­sid­er­ing the fac­tors involved in scaf­fold fab­ri­ca­tion, such as denat­u­ra­tion tem­per­a­ture and issues relat­ed to bio­log­i­cal safe­ty, col­la­gen orig­i­nat­ing from tilapia fish is thought to be a suit­able bio­ma­te­r­i­al to replace mam­malian col­la­gen for use in clin­i­cal regen­er­a­tive med­i­cine. The high­er ther­mal sta­bil­i­ty of tilapia fish col­la­gens com­pared to oth­er marine sources jus­ti­fies its uti­liza­tion in TE.  How­ev­er, fur­ther ani­mal exper­i­ments are need­ed before the col­la­gen can be applied clinically.

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Corresponding author biography

Norhi­dayu Muhamad Zain obtained her Ph.D. from the Uni­ver­si­ti Teknolo­gi Malaysia (UTM) in 2016. Cur­rent­ly, she is a Senior Lec­tur­er at the Fac­ul­ty of Social Sci­ences and Human­i­ties, UTM. Her cur­rent research inter­est includes halal sci­ence, bio­ma­te­ri­als and bio­med­ical engineering.

© 2021 by the authors. This is an open access arti­cle dis­trib­uted under the Cre­ative Com­mons Attri­bu­tion License, which per­mits unre­strict­ed use, dis­tri­b­u­tion, and repro­duc­tion in any medi­um, pro­vid­ed the orig­i­nal work is prop­er­ly cited.

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