Uncultured Autogenous Adipose-derived Regenerative Cells Promote Bone Formation During Distraction Osteogenesis in Rats
Uncultured Autogenous Adipose-derived Regenerative Cells Promote Bone Formation During Distraction Osteogenesis in Rats
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DOI：
10.1007/s11999-014-3608-8
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发表时间：
2014-12-01
影响因子：
4.2
通讯作者：
Tsuchiya, Hiroyuki
中科院分区：
文献类型：
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作者：
Nomura, Issei;Watanabe, Koji;Tsuchiya, Hiroyuki
研究背景脂肪干细胞在体内外均表现出向多种间充质细胞分化的潜能。这些细胞可以容易地从自体脂肪组织中大量分离，并且在没有培养或分化诱导的情况下使用，这可以使它们相对容易地用于临床目的;问题/目的在啮齿类动物模型中进行的牵张成骨动物研究的问题是未培养的脂肪来源的再生细胞（ADRC），其可以容易地从自体脂肪组织中大量分离，并含有几种类型的干细胞和再生细胞，在牵张成骨中促进骨形成。我们使用几种工具进行评估：（1）骨密度的放射学分析;（2）形成的骨痂的组织学分析;（3）生物力学测试;（4）DiI标记（一种用于植入后细胞示踪的膜染色方法）;和（5）实时聚合酶链反应。牵张结束后立即将生理盐水（对照组）、I型胶原凝胶（胶原组）或ADRC和I型胶原凝胶的混合物（ADRC组）注入牵张骨痂内。结果与对照组相比，注射后6周，ADRC组牵张骨痂的骨密度增加了46%（p = 0.003，Cohen's d = 10.2，95%可信区间[CI] +/- 0.180）。与对照组相比，注射后6周，ADRC组的断裂强度增加了66%（p = 0.006，Cohen d = 1.32，95% CI +/- 0.180）。实时逆转录-聚合酶链反应显示，ADRC组的骨痂中骨形态发生蛋白-2（高7.4倍）、血管内皮生长因子A（高6.8倍）和基质细胞衍生因子-1（高4.3倍）水平较高。结论注射ADRCs可促进牵张骨痂成骨分化，其机制与分泌促进成骨或血管生成的体液因子如骨形态发生蛋白2或血管内皮生长因子A有关。
Background Adipose-derived stem cells have recently shown differentiation potential in multiple mesenchymal lineages in vitro and in vivo. These cells can be easily isolated in large amounts from autologous adipose tissue and used without culturing or differentiation induction, which may make them relatively easy to use for clinical purposes; however, their use has not been tested in a distraction osteogenesis model.Question/purposes The question of this animal study in a rodent model of distraction osteogenesis was whether uncultured adipose-derived regenerative cells (ADRCs), which can easily be isolated in large amounts from autologous adipose tissue and contain several types of stem and regenerative cells, promote bone formation in distraction osteogenesis. We evaluated this using several tools: (1) radiographic analysis of bone density; (2) histological analysis of the callus that formed; (3) biomechanical testing; (4) DiI labeling (a method of membrane staining for postimplant celltracing); and (5) real-time polymerase chain reaction.Methods Sixty rats were randomly assigned to three groups. Physiological saline (control group), Type I collagen gel (collagen group), or a mixture of ADRC and Type I collagen gel (ADRC group) was injected into the distracted callus immediately after distraction termination. To a rat femur an external fixator was applied at a rate of 0.8 mm/day for 8 days.Results The bone density of the distracted callus in the ADRC group increased by 46% (p = 0.003, Cohen's d = 10.2, 95% confidence interval [CI] +/- 0.180) compared with the control group at 6 weeks after injection. The fracture strength in the ADRC group increased by 66% (p = 0.006, Cohen's d = 1.32, 95% CI +/- 0.180) compared with the control group at 6 weeks after injection. Real-time reverse transcription-polymerase chain reaction of the distracted callus from the ADRC group had higher levels of bone morphogenetic protein-2 (7.4 times higher), vascular endothelial growth factor A (6.8 times higher), and stromal cell-derived factor-1 (4.3 times higher). Cell labeling in the newly formed bone showed the ADRCs differentiated into osseous tissue at 3 weeks after injection.Conclusions The injection of ADRCs promoted bone formation in the distracted callus and this mechanism involves both osteogenic differentiation and secretion of humoral factors such as bone morphogenetic protein-2 or vascular endothelial growth factor A that promotes osteogenesis or angiogenesis.