Tissue Regeneration Department, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
Biofabrication. 2013 Jun;5(2):025003. doi: 10.1088/1758-5082/5/2/025003. Epub 2013 Feb 27.
Decellularized extracellular matrix (ECM) has recently gained a lot of interest as an instructive biomaterial for regenerative medicine applications. In this study, the ability of adult human mesenchymal stem cell (hMSC)-derived ECM to rescue the phenotype of osteoarthritic (OA) chondrocytes and to further stimulate the differentiation of healthy (HL) chondrocytes was evaluated. ECMs were prepared by decellularizing hMSCs cultured in basic medium (BM) and chondrogenic medium (CM). The obtained ECM was then combined with a polymeric solution of Poly (ε-caprolactone) (PCL) dissolved in 1, 1, 1, 3, 3, 3-hexafluoro-2-propanol (HFIP) and electrospun meshes were fabricated. Electrospun ECM scaffolds were characterized using scanning electron microscopy (SEM) and picrosirius red staining was used to confirm the presence of collagen. OA and HL chondrocytes were cultured on scaffolds containing hMSC ECM in BM or CM and compared to PCL electrospun scaffolds without ECM. Metabolic activity and chondrogenic gene expression were assessed by Alamar blue assay and quantitative PCR (qPCR) analysis, respectively. The ECM presence resulted in a significant difference in chondrocyte metabolic activity compared to PCL scaffolds alone. HL chondrocytes cultured for 21 days in chondrogenic medium on electrospun scaffolds containing hMSC ECM from BM showed a significant increase in collagen II and aggrecan expression compared to hMSC ECM from CM and PCL scaffolds without ECM incorporation. No significant influence of hMSC ECM presence on the chondrogenic signature of OA chondrocytes was found. The influence of decellularized hMSC ECM on HL chondrocytes suggests that hMSC-derived ECM scaffolds are promising candidates for cartilage tissue engineering applications.
去细胞细胞外基质 (ECM) 最近作为再生医学应用的有指导意义的生物材料引起了广泛关注。在这项研究中,评估了成人间充质干细胞 (hMSC) 衍生 ECM 拯救骨关节炎 (OA) 软骨细胞表型的能力,并进一步刺激健康 (HL) 软骨细胞的分化。通过在基础培养基 (BM) 和软骨形成培养基 (CM) 中培养 hMSC 来制备 ECM。然后将获得的 ECM 与溶解在 1,1,1,3,3,3-六氟-2-丙醇 (HFIP) 中的聚 (ε-己内酯) (PCL) 的聚合物溶液结合,并制造出电纺网。使用扫描电子显微镜 (SEM) 对电纺 ECM 支架进行了表征,并使用苦味酸红染色来确认胶原蛋白的存在。OA 和 HL 软骨细胞在 BM 或 CM 中培养在含有 hMSC ECM 的支架上,并与不含 ECM 的 PCL 电纺支架进行比较。通过阿尔玛蓝测定和定量 PCR (qPCR) 分析分别评估代谢活性和软骨生成基因表达。与单独的 PCL 支架相比,ECM 的存在导致软骨细胞代谢活性有显著差异。在含有 BM 来源的 hMSC ECM 的电纺支架上在软骨形成培养基中培养 21 天的 HL 软骨细胞与 CM 来源的 hMSC ECM 和不含 ECM 的 PCL 支架相比,胶原蛋白 II 和聚集蛋白聚糖的表达显著增加。未发现 hMSC ECM 存在对 OA 软骨细胞软骨生成特征有显著影响。去细胞化的 hMSC ECM 对 HL 软骨细胞的影响表明,hMSC 衍生的 ECM 支架是软骨组织工程应用的有前途的候选物。