Decellularized extracellular matrix is one form of natural material in tissue engineering. The process of dECM retains the tissue microstructure, provides good cell adhesion sites, maintains most of biological signals that promotes the survival and differentiation ability of cells. In this study, sheep kidney was decellularized followed by histochemical staining, elemental analysis and scanning electron microscopy characterizations. The dECM scaffold was prepared with different sequences of freeze drying technology, crosslinking and the water absorption, porosity, mechanical strength with subsequent thermogravimetric analysis, Infrared spectroscopy and biocompatibility tests. Our results indicated that these decellularized treatments of sheep kidney can effectively remove DNA and retain uniform pore size distribution. After crosslinking the scaffold’s water absorption decreased from 987.56 ± 40.21% to 934.39 ± 39.61%, the porosity decreased from 89.64 ± 3.2% to 85.09 ± 17.63%, and the compression modulus increased from 304.32 ± 25.43 kPa to 459.53 ± 38.92 kPa, with thermal process the percentage of weight loss decreased from 66.57% to 44.731%, in addition, the composition didn’t change significantly, crosslinking could also promote the stability. In terms of biocompatibility, the number of viable cells increased significantly with the days. In conclusion, the crosslinked decellularized sheep kidney extracellular matrix scaffold reduced water absorption and porosity slightly, but has a significant increase in mechanical properties, and presented excellent biocompatibility which are beneficial to cell adhesion, growth and differentiation.
StendahlJCKaufmanDBStupp. Extracellular matrix in pancreatic islets: relevance to scaffold design and transplantation. Cell Transplant2009; 18(1): 1–12. DOI: 10.3727/096368909788237195.
2.
NabaAClauserKRHoerschS, et al.The matrisome: in silico definition and in vivo characterization by proteomics of normal and tumor extracellular matrices. Molecular & Cellular Proteomics2012; 11(4): M111.014647. DOI: 10.1074/mcp.m111.014647.
KaramanosNKPiperigkouZTheocharisAD, et al.Proteoglycan chemical diversity drives multifunctional cell regulation and therapeutics. Chem Rev2018; 118(18): 9152–9232. DOI: 10.1021/acs.chemrev.8b00354.
5.
GattazzoFUrciuoloABonaldo, Extracellular matrix: a dynamic microenvironment for stem cell niche. Biochimica et Biophysica Acta (BBA) - General Subjects2014; 1840(8): 2506–2519. DOI: 10.1016/j.bbagen.2014.01.010.
6.
LuPFTakaiKWeaverVM. Extracellular matrix degradation and remodeling in development and disease. Cold Spring Harbor Perspectives in Biology2011; 3(12): a005058. DOI: 10.1101/cshperspect.a005058.
7.
KimSHTurnbullJGuimondS, Extracellular matrix and cell signalling: the dynamic cooperation of integrin, proteoglycan and growth factor receptor. J Endocrinol2011; 209(2): 139–151. DOI: 10.1530/joe-10-0377.
8.
ZhaoYGZhuC. Extra cellular matrix and matrix metalloproteinases. Progress in Biochemistry and Biophysics1999; 26(3): 223–228.
9.
BurchPTKazaAKLambertLM, et al.Clinical performance of decellularized cryopreserved valved allografts compared with standard allografts in the right ventricular outflow tract. The Annals of Thoracic Surgery2010; 90(4): 1301–1306. DOI: 10.1016/j.athoracsur.2010.05.024.
10.
BrownBNValentinJEStewart-AkersAM, et al.Macrophage phenotype and remodeling outcomes in response to biologic scaffolds with and without a cellular component. Biomaterials2009; 30(8): 1482–1491. DOI: 10.1016/j.biomaterials.2008.11.040.
11.
ShojaieL. Characterization methods of acellularized tissue and organs. KajbafzadehA. M., Editors 2021. p. 1–6. DOI: 10.1007/978-3-030-82735-9_1.
12.
XuJFangHZhengS, et al.A biological functional hybrid scaffold based on decellularized extracellular matrix/gelatin/chitosan with high biocompatibility and antibacterial activity for skin tissue engineering. Int J Biol Macromol2021; 187: 840–849. DOI: 10.1016/j.ijbiomac.2021.07.162.
13.
OttHCMatthiesenTSGohSK, et al.Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med2008; 14(2): 213–221. DOI: 10.1038/nm1684.
14.
DuJHuXSuY, et al.Gelatin/sodium alginate hydrogel-coated decellularized porcine coronary artery to construct bilayer tissue engineered blood vessels. Int J Biol Macromol2022; 209: 2070–2083. DOI: 10.1016/j.ijbiomac.2022.04.188.
15.
WoodsTGratzerPF. Effectiveness of three extraction techniques in the development of a decellularized bone–anterior cruciate ligament–bone graft. Biomaterials2005; 26(35): 7339–7349. DOI: 10.1016/j.biomaterials.2005.05.066.
16.
UygunBESoto-GutierrezAYagiH, et al.Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med2010; 16(7): 814–820. DOI: 10.1038/nm.2170.
17.
ReingJEBrownBNDalyKA, et al.The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffolds. Biomaterials2010; 31(33): 8626–8633. DOI: 10.1016/j.biomaterials.2010.07.083.
18.
HuangSFuX. Naturally derived materials-based cell and drug delivery systems in skin regeneration. J Control Release2010; 142(2): 149–159. DOI: 10.1016/j.jconrel.2009.10.018
CrapoPMGilbertTWBadylakSF. An overview of tissue and whole organ decellularization processes. Biomaterials2011; 32(12): 3233–3243. DOI: 10.1016/j.biomaterials.2011.01.057.
21.
YiSXuLGuXS. Scaffolds for peripheral nerve repair and reconstruction. Exp Neurol2019; 319: 112761. DOI: 10.1016/j.expneurol.2018.05.016.
22.
LiWHuXWangS, et al.Multiple comparisons of three different sources of biomaterials in the application of tumor tissue engineering in vitro and in vivo. Int J Biol Macromol2019; 130: 166–176. DOI: 10.1016/j.ijbiomac.2019.02.136.
23.
ShiraniAGanjiFGolmohammadiM, et al.Cross-linked acellular lung for application in tissue engineering: effects on biocompatibility, mechanical properties and immunological responses. Mater Sci Eng C Mater Biol Appl2021; 122: 111938. DOI: 10.1016/j.msec.2021.111938.
24.
SinghNKHanWNamSA, et al.Three-dimensional cell-printing of advanced renal tubular tissue analogue. Biomaterials2020; 232: 119734. DOI: 10.1016/j.biomaterials.2019.119734.
25.
GuanYLiuSLiuY, et al.Porcine kidneys as a source of ECM scaffold for kidney regeneration. Mater Sci Eng C Mater Biol Appl2015; 56: 451–456. DOI: 10.1016/j.msec.2015.07.007.
26.
RemuzziA. Experimental evaluation of kidney regeneration by organ scaffold recellularization. Scientific Reports2017; 7. DOI: 10.1038/srep43502.
27.
SullivanDCMirmalek-SaniSHDeeganDB, et al.Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. Biomaterials2012; 33(31): 7756–7764. DOI: 10.1016/j.biomaterials.2012.07.023.
28.
YangSZhengLChenZ, et al.Decellularized pig kidney with a micro-nano secondary structure contributes to tumor progression in 3D tumor model. Materials2022; 15(5): 1935–1935. DOI: 10.3390/ma15051935.
29.
SimonPKasimirMTSeebacherG, et al.Early failure of the tissue engineered porcine heart valve SYNERGRAFT in pediatric patients. Eur J Cardiothorac Surg2003; 23(6): 1002–1006. DOI: 10.1016/s1010-7940(03)00094-0.
30.
ChengCPengXXiL, et al.Feasibility study of oxidized naringin as a novel crosslinking agent for crosslinking decellularized porcine Achilles tendon and its potential application for anterior cruciate ligament repair. J Biomed Mater Res A2023; 111: 170–184. DOI: 10.1002/jbm.a.37440.
31.
FischerAH. Hematoxylin and eosin staining of tissue and cell sections. CSH protocols, 2008: p. pdb.prot4986.
32.
ChenRNHoHOTsaiYT, et al.Process development of an acellular dermal matrix (ADM) for biomedical applications. Biomaterials2004; 25(13): 2679–2686. DOI: 10.1016/j.biomaterials.2003.09.070.
33.
LihEParkKWChunSY, et al.Biomimetic porous PLGA scaffolds incorporating decellularized extracellular matrix for kidney tissue regeneration. ACS Appl Mater Interfaces2016; 8(33): 21145–21154. DOI: 10.1021/acsami.6b03771.
34.
ZhangXXuLHuangX, et al.Structural study and preliminary biological evaluation on the collagen hydrogel crosslinked by γ-irradiation. J Biomed Mater Res A2012; 100(11): 2960–2969. DOI: 10.1002/jbm.a.34243.
35.
PieperJSOosterhofADijkstraPJ, et al.Preparation and characterization of porous crosslinked collagenous matrices containing bioavailable chondroitin sulphate. Biomaterials1999; 20(9): 847–858. DOI: 10.1016/s0142-9612(98)00240-3.
36.
ButtafocoLEngbers-BuijtenhuijsPPootAA, et al.First steps towards tissue engineering of small-diameter blood vessels: Preparation of flat scaffolds of collagen and elastin by means of freeze drying. J Biomed Mater Res B Appl Biomater2006; 77(2): 357–368. DOI: 10.1002/jbm.b.30444.
37.
McKegneyMTaggartIGrantMH. The influence of crosslinking agents and diamines on the pore size, morphology and the biological stability of collagen sponges and their effect on cell penetration through the sponge matrix. J Mater Sci Mater Med2001; 12(9): 833–844. DOI: 10.1023/a:1017989305873.
38.
Olde DaminkLHHDijkstraPJvan LuynMJ, et al.Cross-linking of dermal sheep collagen using a water-soluble carbodiimide. Biomaterials1996; 17(8): 765–773. DOI: 10.1016/0142-9612(96)81413-x.
39.
CaoHXuSY, EDC/NHS-crosslinked type II collagen-chondroitin sulfate scaffold: characterization and in vitro evaluation. J Mater Sci Mater Med2008; 19(2): 567–575. DOI: 10.1007/s10856-007-3281-5.
40.
MagliSRossiLConsentinoC, et al.Combined analytical approaches to standardize and characterize biomaterials formulations: application to chitosan-gelatin cross-linked hydrogels. Biomolecules2021; 11: 683. DOI: 10.3390/biom11050683.
41.
HuangC. Characteristics and preparation of designed alginate-based composite scaffold membranes with decellularized fibrous micro-scaffold structures from porcine skin. Polymers, 2021.
42.
XiaYMittelstaedtDRamakrishnanN, et al.Depth-dependent anisotropies of amides and sugar in perpendicular and parallel sections of articular cartilage by Fourier transform infrared imaging. Microsc Res Tech2011; 74(2): 122–132. DOI: 10.1002/jemt.20881.
43.
VulpeRPopaMPictonL, et al.Crosslinked hydrogels based on biological macromolecules with potential use in skin tissue engineering. Int J Biol Macromol2016; 84: 174–181. DOI: 10.1016/j.ijbiomac.2015.12.019.
44.
OpatrnyK. Clinical importance of biocompatibility and its effect on haemodialysis treatment. Nephrol Dial Transplant2003; 18(Suppl 5): 41–44. DOI: 10.1093/ndt/gfg1044.
