Kamenaga Tomoyuki, Kuroda Yuichi, Nagai Kanto, Tsubosaka Masanori, Takashima Yoshinori, Kikuchi Kenichi, Fujita Masahiro, Ikuta Kemmei, Anjiki Kensuke, Maeda Toshihisa, Nakano Naoki, Takayama Koji, Hashimoto Shingo, Hayashi Shinya, Matsushita Takehiko, Niikura Takahiro, Kuroda Ryosuke, Matsumoto Tomoyuki
Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 7-5-1, Kusunoki-chou, 650-0017, Kobe, Japan.
Stem Cell Res Ther. 2021 Feb 4;12(1):110. doi: 10.1186/s13287-021-02182-3.
BACKGROUND: Novel therapeutic strategies for the healing of nonunion, which has serious effects on the quality of life of patients, are needed. We evaluated the therapeutic effect of local transplantation of human stromal vascular fraction (SVF) cells on fracture healing in a rat non-healing fracture model and compared the effects between freshly isolated (F) and cryopreserved (C)-SVFs. METHODS: Non-healing fracture model was induced in the femur of female immunodeficient rats (F344/N Jcl rnu/rnu) with cauterizing periosteum. Immediately after the creation of non-healing fracture, rats received local transplantation of F and C-SVFs suspended in phosphate-buffered saline (PBS) or the same volume of PBS without cells using the same scaffold as a control group. During 8 weeks post-surgery, radiologic, histological, immunohistochemical, and biomechanical analyses were performed to evaluate fracture healing. The comparison of radiological results was performed with a chi-square test, and the multiple comparisons of immunohistochemical, histological, and biomechanical results among groups were made using a one-way analysis of variance. A probability value of 0.05 was considered to denote statistical significance. RESULTS: At week 8, in 60% of animals receiving F-SVF cells and in 50% of animals receiving C-SVF cells, the fracture radiologically healed with bone union whereas nonunion was observed in the control group. The healing potential was also confirmed by histological and biomechanical assessments. One of the mechanisms underlying healing involving intrinsic angiogenesis/osteogenesis was enhanced in F- and C-SVF groups compared with that in the control group. Human cell-derived vasculogenesis/osteogenesis, which was also confirmed in an in vitro differentiation assay, was also enhanced in the F- and C-SVF groups compared with that in the control groups and could be another mechanism for healing. CONCLUSIONS: SVF cells can enhance bone healing and cryopreserved cells have almost equal potential as fresh cells. SVF cells can be used for improving nonunion bone fracture healing as an alternative to other mesenchymal stem cells and the effect of SVF cells can be maintained under cryopreservation.
背景：骨不连对患者生活质量有严重影响，需要新的治疗策略。我们在大鼠骨不连骨折模型中评估了局部移植人基质血管成分（SVF）细胞对骨折愈合的治疗效果，并比较了新鲜分离（F）和冻存（C）的SVF之间的效果。 方法：通过烧灼骨膜在雌性免疫缺陷大鼠（F344/N Jcl rnu/rnu）的股骨中诱导骨不连骨折模型。在造成骨不连骨折后立即将悬浮于磷酸盐缓冲盐水（PBS）中的F和C - SVF或相同体积不含细胞的PBS作为对照组，使用相同支架进行局部移植。在术后8周内，进行放射学、组织学、免疫组织化学和生物力学分析以评估骨折愈合情况。放射学结果的比较采用卡方检验，组间免疫组织化学、组织学和生物力学结果的多重比较采用单因素方差分析。概率值0.05被认为具有统计学意义。 结果：在第8周时，接受F - SVF细胞的动物中有60%以及接受C - SVF细胞的动物中有50%骨折在放射学上达到骨愈合，而对照组观察到骨不连。组织学和生物力学评估也证实了愈合潜力。与对照组相比，F和C - SVF组中涉及内源性血管生成/骨生成的愈合潜在机制之一增强。在体外分化试验中也得到证实的人细胞源性血管生成/骨生成在F和C - SVF组中也比对照组增强，这可能是另一种愈合机制。 结论：SVF细胞可促进骨愈合，冻存细胞与新鲜细胞具有几乎相同的潜力。SVF细胞可作为其他间充质干细胞的替代物用于改善骨不连骨折愈合，并且SVF细胞的效果在冻存条件下可得以维持。
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