脂肪间充质干细胞植入脱细胞羊膜/聚己内酯组织工程材料的构建与评价  

Fabrication and Evaluation of Decellularized Human Amniotic Membrane/Polycaprolactone Tissue Engineering Materials Implanted with Adipose-Derived Mesenchymal Stem Cells

作　　者：杨德宇 束红梅 刘克海[1] YANG Deyu;SHU Hongmei;LIU Kehai(College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]上海海洋大学食品学院,上海201306

出　　处：《中国细胞生物学学报》2021年第10期1991-2000,共10页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81572989);上海市科委工程中心建设项目(批准号:18430721100)资助的课题。

摘　　要：为探索用于盆底修复的新型生物材料,该研究构建以脱细胞羊膜(decellularized human amniotic membrane,dHAM)为外壳、聚己内酯(polycaprolactone,PCL)为载体,并种植共培养脂肪间充质干细胞(co-culture adipose-derived mesenchymal stem cells,Co-ADSCs)的组织工程材料。首先,经体外分离并鉴定得到脂肪间充质干细胞(adipose-derived mesenchymal stem cells,ADSCs),建立共培养体系,并采用免疫荧光法、qRT-PCR和ELISA检测Co-ADSCs合成I型胶原蛋白的能力。同时,制备dHAM并判断脱细胞程度,用溶液浇铸法制备PCL薄膜,将dHAM复合于PCL薄膜两侧,并进行8周体外降解实验。最后,将细胞种植于dHAM/PCL,通过SEM、CCK-8、拉伸试验评价材料的细胞相容性和力学性能。结果表明,脂肪源分离的细胞具备间充质干细胞特性,ADSCs经共培养可合成较多I型胶原蛋白。制备的复合材料为"三明治"结构,降解8周后材料的剩余量为82.12%。dHAM/PCL细胞相容性好,且种植Co-ADSCs提升了材料的力学性能。综上,基于组织工程原理构建的Co-ADSCs-dHAM/PCL复合生物材料可降解、细胞相容且机械性能好,有望作为新型盆底修复材料。In order to explore a new biomaterial for pelvic floor repair,a tissue engineering material with Co-ADSCs(co-culture adipose-derived mesenchymal stem cells)was constructed using dHAM(decellularized human amniotic membrane)as coat and PCL(polycaprolactone)as carrier.First,ADSCs(adipose-derived mesenchymal stem cells)were isolated and identified in vitro,and a co-culture system was established.Immunofluorescence,qRT-PCR and ELISA were used to detect the ability of Co-ADSCs to synthesize type I collagen.Meanwhile,dHAM was prepared and the degree of decellularization was evaluated.PCL film was prepared by solution casting method,and dHAM was composite on both sides of PCL film.The degradation test was carried out in vitro for 8 weeks.Finally,the cells were planted in dHAM/PCL,and the cytocompatibility and mechanical properties of the materials were evaluated by SEM,CCK-8 and tensile test.The results showed that adipogenic cells had the characteristics of mesenchymal stem cells,and ADSCs produced more type I collagen by co-culture.The prepared composite was a“sandwich structure”,and the residual content reached 82.12%after 8 weeks of degradation.The dHAM/PCL cell compatibility was good,and the mechanical parameters of the materials were improved after Co-ADSCs transplantation.In conclusion,the tissue-engineered Co-ADSCs-dHAM/PCL composite biomaterial is degradable,cell-compatible and has good mechanical properties,so it is expected to be a new pelvic floor repair material.

关 键 词：脂肪间充质干细胞 脱细胞羊膜 聚己内酯 组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]