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首页> 外文期刊>Biomaterials >In vitro and in vivo evaluation of differentially demineralized cancellous bone scaffolds combined with human bone marrow stromal cells for tissue engineering.
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In vitro and in vivo evaluation of differentially demineralized cancellous bone scaffolds combined with human bone marrow stromal cells for tissue engineering.

机译：体外和体内差异脱矿质松质骨支架与人骨髓基质细胞结合进行组织工程评估。

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Mineralized and partially or fully demineralized biomaterials derived from bovine bone matrix were evaluated for their ability to support human bone marrow stromal cell (BMSC) osteogenic differentiation in vitro and bone-forming capacity in vivo in order to assess their potential use in clinical tissue-engineering strategies. BMSCs were either seeded on bone-derived scaffolds and cocultured in direct cell-to-scaffold contact, allowing for the exposure of soluble and insoluble matrix-incorporated factors, or cocultured with the scaffold preparations in a transwell system, exposing them to soluble matrix-incorporated factors alone. Osteoblast-related markers, alkaline phosphatase (ALP) activity and bone sialoprotein (BSP) and osteopontin (OP) mRNA expression were evaluated in BMSCs following 14 days of cocultivation in both systems. The data demonstrate that BMSCs from some donors express significantly higher levels of all osteoblast-related markers following cocultivation in direct cell-to-scaffold contact with mineralized scaffolds in comparison to fully demineralized preparations, while BMSCs from other donors display no significant differences in response to various scaffold preparations. In contrast, BMSCs cocultured independently with soluble matrix-incorporated factors derived from each scaffold preparation displayed significantly lower levels of ALP activity and BSP mRNA expression in comparison to untreated controls, while no significant differences were observed in marker levels between cells cocultured similarly with different biomaterial preparations. In addition, BMSCs were seeded directly on mineralized and partially or fully demineralized biomaterials and implanted in subcutaneous sites of athymic mice for 8 weeks to evaluate their in vivo bone-forming capacity. The ex vivo incorporation of BMSCs into all bone-derived scaffold preparations substantially increased the mean extent and frequency of samples containing de novo bone formation over similar nonseeded controls, as determined by histological and histomorphometrical analysis. No statistically significant differences were observed in the extent or frequency of bone formation between various scaffold preparations seeded with BMSCs from different donors. These results demonstrate that the in vivo osteoinductivity of bone-derived scaffolds can be modulated by ex vivo incorporated BMSCs and the extent of scaffold demineralization plays a significant role in influencing in vitro osteogenic differentiation of BMSCs depending on the coculture system and BMSC donor.
机译：评价了源自牛骨基质的矿化和部分或完全脱矿物质的生物材料在体外支持人骨髓基质细胞（BMSC）成骨分化的能力以及体内成骨的能力，以评估其在临床组织工程中的潜在用途策略。将BMSCs接种在骨源性支架上并在直接的细胞与支架接触中共培养，以暴露可溶和不溶性基质结合因子，或者将其与支架制备物一起在Transwell系统中共培养，将其暴露于可溶性基质单独考虑因素。在两个系统中共培养14天后，在BMSC中评估成骨细胞相关标志物，碱性磷酸酶（ALP）活性以及骨唾液蛋白（BSP）和骨桥蛋白（OP）mRNA表达。数据表明，与完全脱矿质的制剂相比，在与矿化的支架直接细胞接触的支架中共培养后，来自一些供体的BMSC表达出明显更高的所有成骨细胞相关标志物，而来自其他供体的BMSC在反应方面无显着差异。各种脚手架的准备。相比之下，与未处理的对照组相比，与来自每种支架制备物的可溶性基质结合因子独立共培养的BMSC与未处理的对照组相比，其ALP活性和BSP mRNA表达水平显着降低，而在与不同生物材料相似共培养的细胞之间，标志物水平没有观察到显着差异准备。另外，将BMSC直接接种在矿化的和部分或完全脱矿物质的生物材料上，并植入无胸腺小鼠的皮下部位8周，以评估其体内成骨能力。如通过组织学和组织形态计量学分析所确定的，将BMSC离体结合到所有骨源性支架制剂中，与相似的非播种对照相比，实质上增加了包含新生骨形成的样品的平均程度和频率。在用来自不同供体的BMSCs接种的各种支架制剂之间，在骨形成的程度或频率上没有观察到统计学上的显着差异。这些结果表明，可以通过离体结合的BMSCs来调节骨源性支架的体内骨诱导性，并且支架的去矿质化程度在影响BMSCs的体外成骨分化中起着重要作用，这取决于共培养系统和BMSC供体。

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