软骨脱细胞基质仿生支架的制备及其对骨髓基质干细胞成软骨分化的影响

目的探讨软骨脱细胞基质(ACM)仿生支架的制备及其对骨髓基质干细胞(BMSCs)成软骨分化的影响。方法将软骨组织粉碎后脱细胞处理,并以不同的比例与明胶(GT)溶液混合并交联,冷冻干燥制成多孔仿生支架。用扫描电镜(SEM)观察不同交联比例制成的ACM/GT支架,并对其孔径、孔隙率、生物力学、降解速率进行评估,从而选取最优组用于体外软骨构建。分离并培养山羊BMSCs,接种于单纯GT和ACM/GT支架上,SEM观察培养1、7、14 d后细胞在支架上的黏附、分布与基质分泌情况,并通过实时定量聚合酶链反应(qRT-PCR)评估成软骨分化能力。结果制备的ACM无细胞残留。随着ACM占比增加,孔径逐渐增大,降解速率逐渐增快,力学强度逐渐降低。其中G2A2组在孔径、孔隙率、生物力学和降解速率方面均适宜体外构建软骨组织。SEM显示细胞在单纯GT和G2A2支架上均匀分布,增殖显著,基质分泌明显。qRT-PCR显示G2A2显著促进了BMSCs的成软骨分化。结论ACM可以制备成适宜软骨再生的支架材料,并可促进BMSCs的成软骨分化。

Fabrication of Acellular Cartilage Matrix Biomimetic Scaffolds and Their Effects on Chondrogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

Objective To explore the fabrication of acellular cartilage matrix-derived(ACM)biomimetic scaffolds and their effects on chondrogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods The cartilage tissue was decellularized after pulverization,and then mixed and crosslinked with gelatin(GT)solution in different proportions.The mixture was then freeze-dried to make porous biomimetic scaffolds.Scanning electron microscopy(SEM)was used to observe the microstructure of ACM/GT scaffolds with different proportions.The pore size,porosity,biomechanics and degradation rate of scaffolds were evaluated to select the optimal proportion for cartilage construction in vitro.Goat BMSCs were cultured and seeded onto pure GT and ACM/GT scaffolds.The adhesion,distribution and matrix secretion of cells on the scaffolds were observed by SEM after 1,7 and 14 days of culture,and the chondrogenic ability was evaluated by quantitative real-time polymerase chain reaction(qRT-PCR).Results The cell fragments were completely removed in ACM.With the increase of ACM content,the pore size and degradation rate increased while the mechanical strength decreased gradually.In terms of the pore size,porosity,biomechanics and degradation rate,the G2A2 group was suitable for cartilage construction in vitro.SEM showed that the cells were evenly distributed on the pure GT and G2A2 scaffolds,with significant proliferation and matrix secretion.qRT-PCR showed that G2A2 significantly promoted the chondrogenic differentiation of BMSCs.Conclusion ACM can be used to fabricate biomimetic scaffolds for cartilage regeneration,and ACM can promote the chondrogenic differentiation of BMSCs.