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脂肪源性干细胞表达骨形态发生蛋白 2 与 nHA/RHLC/PLA 支架联合用于大节段桡骨缺损中的增强骨形成。

Enhanced bone formation in large segmental radial defects by combining adipose-derived stem cells expressing bone morphogenetic protein 2 with nHA/RHLC/PLA scaffold.

机构信息

Department of Orthopaedics & Traumatology, Provincial Hospital affiliated to Shandong University, Ji'nan, People's Republic of China.

出版信息

Int Orthop. 2010 Dec;34(8):1341-9. doi: 10.1007/s00264-009-0946-3. Epub 2010 Feb 7.

Abstract

In this study, rabbit adipose-derived stem cells (rASCs) were isolated, cultured in vitro, and transfected with recombinant adenovirus vector containing human bone morphogenetic protein 2 (Ad-hBMP2). These cells were combined with a nano-hydroxyapatite/recombinant human-like collagen/poly(lactic acid) scaffold (nHA/RHLC/PLA) to fabricate a new biocomposite (hBMP2/rASCs-nHA/RHLC/PLA, group 1) and cultured in osteogenic medium. Non-transfected rASCs mixed with nHA/RHLC/PLA (rASCs-nHA/RHLC/PLA, group 2) and nHA/RHLC/PLA scaffold alone (group 3) served as controls. Scanning electron microscope (SEM) demonstrated integration of rASCs with the nHA/RHLC/PLA scaffold. Quantitative real-time RT-PCR analyses of collagen I, osteonectin, and osteopontin cDNA expression indicated that the osteogenic potency of rASCs was enhanced by transfection with Ad-hBMP2. After in vitro culture for seven days, three groups were implanted into 15-mm length critical-sized segmental radial defects in rabbits. After 12 weeks, radiographic and histological analyses were performed. In group 1, the medullary cavity was recanalised, bone was rebuilt and moulding was finished, the bone contour had begun to remodel and scaffold was degraded completely. In contrast, bone defects were not repaired in groups 2 or 3. Furthermore, the scaffold degradation rate in group 1 was significantly higher than in groups 2 or 3. In summary, after transduction with Ad-hBMP2, the osteogenesis of rASCs was enhanced; a new biocomposite created with these cells induced repair of a critical bone defect in vivo in a relatively short time.

摘要

在这项研究中,我们分离并体外培养了兔脂肪来源的干细胞(rASCs),然后用含有人骨形态发生蛋白 2(Ad-hBMP2)的重组腺病毒载体转染这些细胞。将这些细胞与纳米羟基磷灰石/重组人胶原蛋白/聚乳酸(nHA/RHLC/PLA)支架结合,构建了一种新的生物复合材料(hBMP2/rASCs-nHA/RHLC/PLA,第 1 组),并在成骨培养基中培养。未转染的 rASCs 与 nHA/RHLC/PLA(rASCs-nHA/RHLC/PLA,第 2 组)和 nHA/RHLC/PLA 支架单独混合(第 3 组)作为对照。扫描电子显微镜(SEM)显示 rASCs 与 nHA/RHLC/PLA 支架整合。对 I 型胶原、骨连蛋白和骨桥蛋白 cDNA 表达的定量实时 RT-PCR 分析表明,Ad-hBMP2 转染增强了 rASCs 的成骨能力。体外培养 7 天后,将三组分别植入兔 15mm 长的临界尺寸节段性桡骨缺损。12 周后,进行影像学和组织学分析。在第 1 组中,骨髓腔再通,骨重建和塑形完成,骨轮廓开始重塑,支架完全降解。相比之下,第 2 组和第 3 组的骨缺损未得到修复。此外,第 1 组的支架降解率明显高于第 2 组或第 3 组。综上所述,rASCs 经 Ad-hBMP2 转导后,成骨能力增强;用这些细胞构建的新型生物复合材料在体内短时间内诱导了临界骨缺损的修复。

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