Comparison Of Three Skin Tissue Engineering Scaffold Materials

Objective:1?Bone marrow mesenchymal stem cells(BMSCs)were isolated and BMSCs surface markers were identified by flow cytometry.2 ? Preparation of human-derived amniotic membrane,sub-amniotic membrane,umbilical cord acellular tissue scaffold3?By comparing the pros and cons of the relevant properties of the three acellular tissue scaffolds,one is selected as a scaffold material for tissue engineering skin.Methods:1?BMSCs were isolated from SD rat by whole bone marrow adherence2?BMSCs surface markers were identified by flow cytometry.3?The differentiated potential of BMSCs was identified by adipogenic and osteogenic differentiation.4?The umbilical cord arteries and sub-amniotic acellular tissue scaffolds were prepared by SDS-nuclease digestion;the amniotic acellular tissue scaffolds were prepared by TritonX-100 and trypsin digestion.5?The decellularization of the scaffold was observed by hematoxylin and eosin(HE)staining.6?The surface condition of the tissue before and after decellularization was observed by scanning electron microscope.7?The mechanical strength and compliance of the acellular tissues were detected by a universal tester.8?The toxicity of acellular stent to cells was detected by Cell Counting Kit-8(CCK-8).9?The isolated BMSCs cells were used as seed cells,and three kinds of acellular tissue scaffolds were co-cultured in vitro to prepare three kinds of biological dressings with the cells.Results:1?BMSCs were homogeneous in morphology and arranged neatly.The purity of BMSCs was high.The rates of positive markers CD29 and CD90 were(99.8 ± 0.15)% and(96.8 ± 0.89)%.The rate of negative marker CD45 was(2.2 ± 1.23)%2?Calcium nodules induced by osteogenesis differentiation and lipid droplets induced by adipogenic differentiation were showed.3?After acellular treatment,the amniotic membrane,sub-amniotic membrane,and umbilical cord arteries were stained with HE to demonstrate that the surface was amorphous,and the structure of collagen fibers was complete without cell residue.4?Scanning electron microscopy revealed that the three tissues had a rough surface after decellularization.The pore size was altered,and there was no cell residue.5?After testing by universal tester,the maximum pulling force of the amniotic membrane,sub-amniotic membrane,and umbilical artery was(4.56±0.21)N,(13.46±0.18)N,(7.81±0.25)N,and the sub-amnion and amniotic membrane were obtained after decellularization.The maximum tensile lengths of umbilical cord arteries were(5.6±0.24)mm,(4.07±0.25)mm,(2.43±0.18)mm,and the mechanical strength of the sub-amniotic layer was the highest among the three decellularized scaffolds.6?After the three decellularized scaffolds were combined with the cells,the adherence of the cells to the tissues was observed by light microscopy,and the proliferation rate of the BMSCs in the tissues was similar to the proliferation rate of the BMSCs in the culture flasks.7?The cytotoxicity of the three acellular tissue scaffolds determined by the CCK-8 assay was classified as Grade 1,with relatively low toxicity.Conclusion:1?BMSCs obtained by whole bone marrow adherent separation culture had high purity and good condition.2?Umbilical artery and sub-amniotic membrane were treated with SDS-nuclease decellularization and amniotic membrane was treated with TritonX-100 and trypsin to obtain the ideal acellular tissue scaffold.3?The co-culture of BMSCs and three acellular tissue scaffolds can produce ideal results.Mechanical characteristics and biological activity of biological dressings,three kinds of biological dressings in the sub-umbilical amniotic membrane is more suitable for skin tissue engineering stents.