观察松质骨BMG作为组织工程软骨载体的可行性和有效性

摘要

Objective To study the feasibility and effectiveness of cancellous bone BMG as a tissue engineered cartilage carrier.Methods Allogeneic BMG was prepared by sequential defatting,demineralization and denaturation.Isolated rabbit chondrocytes were seeded onto allogeneic cancellous BMG,and cell-BMG constructs were harvested after 1,4 and 7 weeks for evaluation using hematoxylin and eosin staining for overall morphology,toluidine blue staining for extracellular matrix proteoglycans,immunohistochemical staining for collagen type Ⅱ and transmission electron microscope for the examination of cellular microstructure on BMG.Results The prepared BMG was highly porous with mechanical strength adjustable by duration of demineralization and could be easily trimmed for tissue repair.At 1,4 and 7 weeks,the percentage of newly formed cartilage tissue was 11.43 ± 2.16,27.41 ± 4.42 and 44.89 ± 6.05,respectively,and the differences were statistically significant(P＜0.05).The average gray values of type Ⅱ collagen were 153.62 ± 6.37,135.49 ± 7.05 and 120.18 ± 5.34,respectively,and the differences were statistically significant(P ＜ 0.05).The average gray value of proteoglycan was 144.69 ± 4.05,120.37 ± 2.85 and 101.96 ± 3.52,respectively,and the differences were statistically significant(P ＜0.05).Conclusion The cancellous bone BMG has good biocompatibility,adjustable mechanical strength and degradation rate,weak immunogenicity,high availability and suitable porosity.%目的 探索松质骨骨基质明胶(bonematrixgelatin,BMG)作为组织工程软骨载体的可行性和有效性.方法 取新西兰白兔肱骨,通过脱脂、去矿化和去蛋白质多糖制备松质骨BMG.将分离的兔软骨细胞接种到松质骨BMG上,1、4、7 w后收获细胞BMG构建体,通过苏木精和伊红染色评价整体形态,甲苯胺蓝细胞外基质(ECM)蛋白聚糖,免疫组化染色胶原型Ⅱ和扫描电子显微镜观察BMG上的细胞微结构.结果 制备的BMG是高度多孔的,并且容易修剪各种形状用于组织修复.培养时间为1、4、7 w,新形成的软骨组织百分比分别为(11.43 ± 2.16)%、(27.41 ± 4.42)%和(44.89 ± 6.05)%,差异有统计学意义(P ＜0.05);Ⅱ型胶原蛋白的平均灰度值分别为153.62 ± 6.37、135.49 ± 7.05和120.18 ± 5.34,差异有统计学意义(P ＜0.05);蛋白聚糖的平均灰度值分别为144.69 ± 4.05、120.37 ± 2.85、101.96 ± 3.52,差异有统计学意义(P ＜0.05).结论 松质骨BMG具有良好的生物相容性,可控的机械强度和降解速率,显著的低免疫原性,高可用性和合适的孔隙率.