Abstract
The design of composite tissue scaffolds containing an extracellular matrix (ECM) and synthetic polymer fibers is a new approach to create bioactive scaffolds that can enhance cell function. Currently, studies investigating the effects of ECM-deposition and decellularization on polymer degradation are still lacking, as are data on optimizing the stability of the ECM-containing composite scaffolds during prolonged cell culture. In this study, we develop fibrous scaffolds using three polymer compositions, representing slow (E0000), medium (E0500), and fast (E1000) degrading materials, to investigate the stability, degradation, and mechanics of the scaffolds during ECM deposition and decellularization, and during the complete cellularization-decell-recell cycle. We report data on percent molecular weight (% Mw) retention of polymeric fiber mats, changes in scaffold stiffness, ECM deposition, and the presence of fibronectin after decellularization. We concluded that the fast degrading E1000 (Mw retention ≤ 50% after 28 days) was not sufficiently stable to allow scaffold handling after 28 days in culture, while the slow degradation of E0000 (Mw retention ≥ 80% in 28 days) did not allow deposited ECM to replace the polymer support. The scaffolds made from medium degrading E0500 (Mw retention about 60% at 28 days) allowed the gradual replacement of the polymer network with cell-derived ECM while maintaining the polymer network support. Thus, polymers with an intermediate rate of degradation, maintaining good scaffold handling properties after 28 days in culture, seem best suited for creating ECM-polymer composite scaffolds.
Highlights
Extracellular matrix (ECM) is a cell-derived fibrous network, composed of proteins and glycosaminoglycans (GAGs), which displays complex combinations of biochemical, mechanical, and topographical cues
Our results support the conclusions that, for the polymers tested here, (i) cell culture conditions and decellularization do not affect polymer degradation or fiber morphology under all test conditions; (ii) polymer degradation does not affect fiber morphology, even after 28 days; and (iii) cell culture time has the greatest impact on fiber mat stability
We developed extracellular matrix (ECM)-polymer composite scaffolds using three selected polymer compositions with slow (E0000), medium (E0500), and fast (E1000) rates of degradation
Summary
Extracellular matrix (ECM) is a cell-derived fibrous network, composed of proteins and glycosaminoglycans (GAGs), which displays complex combinations of biochemical, mechanical, and topographical cues. A less common, but increasingly investigated approach is to use cells to deposit their native ECM onto the scaffold, followed by decellularization and the use of this ECM-polymer composite scaffold as a bioactive support for subsequent recellularization [33,34,35]. Despite the increasing interest in ECM-polymer composite scaffolds, studies investigating the effects of ECM-deposition and decellularization on polymer degradation, and the stability of the ECM-polymer composite scaffolds in prolonged culture conditions are still lacking. This makes it difficult to select polymers rationally and to optimize scaffold properties. FFiigguurree 11.. ((AA)) CChheemmiiccaallccoommppoossitiitoionnofofthtehetetsetsptoplyomlyemrse.rsT.hTehfereferecaercbaorxbyolxicyaliccidacgirdougprooufpthoef DthTeuDnTit uisnmit aisrkmeadrkinedreind rfeodr fcolarrciltayr.itWy.eWueseudsetdhethneontaottiaotnioEnXEXX0X00t0otonanmame eththeseeseppoolylymmeerrss,,wwiitthhEE ddeennoottiinngg tthhee eetthhyyll eesstteerr aanndd XXXX sshhoowwiinngg tthhee mmoollee ppeerrcceenntt ooff tthhee DDTT.. ((ii)) EE00000000 ddeennootteess 00 mmooll%% ooff DDTT;; ((iiii)) EE00550000 ddeennootteess 55 mmooll%% ooff DDTT;;aanndd((iiiiii))EE11000000ddeennootetess1100mmool%l%pperecrecnent ot foDf DT.TT. hTehteottoatlaalmamouonutnotfoDfTDpTrpesreenset nint itnhetphoelypmoleyrmbaecrkbbaocnkebiosntheeiosntlhyedioffnelryendceiffbeertewneceenbtehtewdeieffnerethnet pdoilfyfemreenrst; apnodly(mB)erSsE;Manmdic(rBo)grSaEpMhs mofieclreocgtrroapsphusnofif beleercmtraotsspmuandfeiboefr(mi)aEt0s0m00a;d(eii)oEf0(5i)0E0;0a0n0d0;((iiiii)) EE10050000.; Tanhde s(ciiai)ffEol1d0s0h0.aTdhcelosscealyff-omldastchhaedd cfilboesreldyi-ammaetctehresdafnidbesrimdiialamremteorrspahnodlosgimy. ilar morphology
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