Abstract
Biologic scaffolds composed of extracellular matrix components have been proposed to repair and reconstruct a variety of tissues in clinical and pre-clinical studies. Injectable gels can fill and conform any three-dimensional shape and can be delivered to sites of interest by minimally invasive techniques. In this study, a biological gel was produced from a decellularized porcine urinary bladder by enzymatic digestion with pepsin. The enzymatic digestion was confirmed by visual inspection after dissolution in phosphate-buffered saline solution and Fourier-transform infrared spectroscopy. The rheological and biological properties of the gel were characterized and compared to those of the MatrigelTM chosen as a reference material. The storage modulus G’ reached 19.4 ± 3.7 Pa for the 30 mg/mL digested decellularized bladder gels after ca. 3 h at 37 °C. The results show that the gel formed of the porcine urinary bladder favored the spontaneous differentiation of human and rabbit adipose-derived stem cells in vitro into smooth muscle cells to the detriment of cell proliferation. The results support the potential of the developed injectable gel for tissue engineering applications to reconstruct for instance the detrusor muscle part of the human urinary bladder.
Highlights
Stem-cell therapies have been shown to be a promising approach to enhance the regeneration of a tissue-engineered bladder [1,2]
Gels derived from the porcine urinary bladder matrix (UBM) produced by pepsin-solubilization have been shown to retain the inherent bioactivity of the matrix with the ability to support adhesion and growth of SMCs when cultured under static in vitro conditions promoting constructive remodeling in tissue engineering [35]
The gel was produced with pepsin-digested decellularized bladder powder (DDB) dissolved in PBS
Summary
Stem-cell therapies have been shown to be a promising approach to enhance the regeneration of a tissue-engineered bladder [1,2]. ECM decellularized scaffolds retain the nanostructure and bioinductive properties of the native matrix, and they have been shown to promote the in vivo creation of site-specific, functional tissue because of the preservation of the structural and functional proteins of the naïve tissue such as glycosaminoglycans, proteoglycans and growth factors [34] In this regard, gels derived from the porcine urinary bladder matrix (UBM) produced by pepsin-solubilization have been shown to retain the inherent bioactivity of the matrix with the ability to support adhesion and growth of SMCs when cultured under static in vitro conditions promoting constructive remodeling in tissue engineering [35]. FTThIeRasmpiedcetrIuamn.dTIhIubsa,ntdhse aarme itdweoImbaanjodr,baasnsodcsiaotfetdhewpitrhottehine FCT=IOR sstpreectctrhuinmg. vTihburast,itohne aanmdiddeirIebctalnydr,ealassteodciatotetdhewpitohlythpeepCt=idOe sbtarecktcbhoinnge cvoibnrfaotrimonatainodn d[4i5re],ctslhyorwelsatsetrdotnogthpeeapkoslyapt e1p64ti6daenbdac1k6b3o5ncemc−o1nfoforrtmheatDioBna[n45d],DsDhoBwssamstrpolnesg, preesapkescattiv1e6l4y6. aTnhdis16b3a5ncdmi−s1afosrenthseitDivBe amnadrDkeDrBosfapmepplteids,eressepceocntdivaerlyy. sTtrhuisctbuarned[i4s5a] saenndsitiitvheams abrekeenr orefppoerptetiddethsaetcoansdhaifrtyosftrtuhcetuamrei[n4e5]I apnedakitthoalsobweeernwreapvoerntuedmtbheartsacasnhifbteorfetlhaeteadmtoinae Idpeceraekastoe lionwtheer wmaovleecnuulmarboerrds ecra,nsubgegreesltaitnegd tthoeapdaerctiraelarseeminovthale omf otleelcoupleaprtoidredserb, ysupgegpessitnindgutrhinegpathrteiaelnrzeymmoavtaicl odfigteeslotipoenpmtidigehstblyeapdetpostihnedluosrisnogf trheeacetnivzeyammaitnicodaicgidesst[i4o7n].mTihgehat mleiadde tIoIItbhaenldosast o12f 3r4eaccmti−v1eshaomwineod amciudcsh[4m7o].rTehreelaamtivideeinIItIebnasnitdy aint 1t2h3e4pcemp−s1ins-hdoiwgeesdtemdugcehl mthoarneirneltahteivneoinnt-ednigsietsyteindtbhleapdedpesrinp-odwigdeesrteidn ggeolotdhaangirneetmheennot nw-ditihgetshteedobblsaedrdveerdpcohwandgere ionfgsoooldubaiglirteyemofenDtDwBitwh tihthe roebsspeervcteeddcthoanDgBe o(Ffisgoulurebi1liat)y. oFfuDrtDheBrmwiotrher,ebspoethctesdamtopDleBs(Fsihgouwreed1a)v. eFruyrtdhiefrfmeroenret, bfiontghesrapmripnltesresghioownesd(fvreormy d1iff0e0r0entot fi4n0g0ercpmrin−1t) rpergoivoindsin(fgroomth1e0r 0s0igtnos4o0f0scumcc−e1s)spfurol venidziynmg aottihcedrisgiegsntisoonf. successful enzymatic digestion
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