University Hospital BG Bergmannsheil Bochum, Bürkle-de-la-Camp Platz 1, 44789, Bochum, Germany.
Deutsches Institut für Zell- und Gewebeersatz, Berlin, Germany.
J Mol Med (Berl). 2019 Oct;97(10):1439-1450. doi: 10.1007/s00109-019-01822-9. Epub 2019 Jul 31.
Critical sized defects, especially in long bones, pose one of the biggest problems in orthopedic surgery. By definition, these defects do not heal without further treatment. Different therapeutic options range from autologous bone grafts, for example, free vascularized bone grafts, to commercially available bone allografts. Disadvantages of these bone allografts are related to reduced osteogenesis, since they are solely composed of cell-free bone matrix. The purpose of this study was to investigate the cell seeding efficiency of human adipose-derived stem cells (hASCs) on human bone allografts in vitro and furthermore analyze these optimized seeded allografts in a critical sized defect model in vivo. Cancellous human bone allografts were colonized with human ASCs in vitro. Cell seeding efficiency was evaluated by Cell Counting Kit-8 assay. Thereafter, optimized hASC-seeded bone scaffolds were examined in a murine femur defect model, stabilized with the MouseExFix system. Subsequently, x-ray analysis and histology were performed. Examination of cell seeding efficiency revealed an optimum starting population of 84,600 cells per 100 mm scaffold. In addition, scaffolds seeded with hASCs showed increased osteogenesis compared with controls. Histological analysis revealed increased remodeling and elevated new bone formation within hASC-seeded scaffolds. Moreover, immunohistochemical stainings revealed increased proliferation, osteogenesis, and angiogenesis. In this study, we systemically optimized cell/volume ratio of two promising components of tissue engineering: hASCs and human bone allografts. These findings may serve as a basis for future translational studies. KEY MESSAGES: Bone tissue engineering. Mesenchymal stem cells derived from human adipose tissue (hASCs). Optimal cell/volume ratio of cell-seeded scaffolds. Increased osteogenesis and angiogenesis in vivo.
临界尺寸缺陷，特别是在长骨中，是骨科手术中最大的问题之一。根据定义，这些缺陷在没有进一步治疗的情况下不会愈合。不同的治疗选择范围从自体骨移植物，例如带血管化骨移植物，到商业上可用的同种异体骨移植物。这些同种异体骨移植物的缺点与成骨减少有关，因为它们仅由无细胞的骨基质组成。本研究的目的是体外研究人脂肪源性干细胞（hASC）在人同种异体骨上的细胞接种效率，并进一步分析这些优化的接种同种异体骨在体内临界尺寸缺陷模型中的情况。体外用人 ASC 殖民同种异体松质骨。通过 Cell Counting Kit-8 测定评估细胞接种效率。此后，用 MouseExFix 系统稳定的小鼠股骨缺损模型检查优化后的 hASC 接种骨支架。随后进行 X 射线分析和组织学检查。细胞接种效率检查显示，每个 100mm 支架的最佳起始细胞数为 84600 个。此外，与对照组相比，接种 hASC 的支架显示出增加的成骨作用。组织学分析显示，接种 hASC 的支架内的重塑和新骨形成增加。此外，免疫组织化学染色显示增殖、成骨和血管生成增加。在这项研究中，我们系统地优化了组织工程中两种有前途的成分的细胞/体积比：hASC 和人同种异体骨。这些发现可能为未来的转化研究提供基础。 关键信息：骨组织工程。来源于人脂肪组织的间充质干细胞（hASC）。细胞接种支架的最佳细胞/体积比。体内成骨和血管生成增加。
