Rust P A, Kalsi P, Briggs T W R, Cannon S R, Blunn G W
Royal National Orthopaedic Hospital, Stanmore, Middlesex, UK.
Clin Orthop Relat Res. 2007 Apr;457:220-6. doi: 10.1097/BLO.0b013e31802e7e8f.
Tissue engineering approaches for bone blocks previously have used synthetic scaffolds. Bone graft (allograft) is used to fill bone defects, but standard processing can lessen this scaffold's osteoinductive potential. We wanted to test if allografts could be used to produce a viable bone block using mesenchymal stem cells. We hypothesized that mesenchymal stem cells differentiate into osteoblasts producing extracellular matrix when cultured on allografts. We also hypothesized that the addition of osteogenic supplements would increase the rate of differentiation. To test these hypotheses, mesenchymal stem cells were isolated from bone marrow aspirated from 10 patients and cultured on allografts from five donors (Group 2), producing 50 samples. This was repeated on allografts heat-treated to denature bioactive proteins (Group 1), and repeated again on allografts to which osteogenic supplements (Group 3) were added. Group 2 mesenchymal stem cells differentiated into osteoblasts producing higher levels of alkaline phosphatase, osteopontin, and Type I collagen matrix protein than Group 1. The rate of differentiation of Group 3 mesenchymal stem cells increased with the supplements. Overall, it was established that the bioactive proteins in the allograft stimulated mesenchymal stem cell differentiation into osteoblasts, with production of extracellular matrix, and that this differentiation increased with the addition of osteogenic supplements.
以往用于骨块的组织工程方法使用的是合成支架。骨移植(同种异体移植)用于填充骨缺损,但标准处理会降低这种支架的骨诱导潜能。我们想测试同种异体移植物是否可用于利用间充质干细胞产生有活力的骨块。我们假设间充质干细胞在同种异体移植物上培养时会分化为产生细胞外基质的成骨细胞。我们还假设添加成骨补充剂会提高分化速率。为了验证这些假设,从10名患者抽取的骨髓中分离出间充质干细胞,并在来自5名供体的同种异体移植物上培养(第2组),共产生50个样本。在经热处理使生物活性蛋白变性的同种异体移植物上重复此操作(第1组),并在添加了成骨补充剂的同种异体移植物上再次重复(第3组)。与第1组相比,第2组间充质干细胞分化为产生更高水平碱性磷酸酶、骨桥蛋白和I型胶原基质蛋白的成骨细胞。第3组间充质干细胞的分化速率随着补充剂而增加。总体而言,已证实同种异体移植物中的生物活性蛋白刺激间充质干细胞分化为产生细胞外基质的成骨细胞,并且这种分化随着成骨补充剂的添加而增加。