( }: y5 j- ?7 l 3基因技术在成骨方面的应用: B% J1 x9 T h* A0 [2 g
7 H K: ]6 D6 H/ b( |& j4 A近年来多种骨生长因子调控基因分离成功及成骨机制的分子水平研究的不断深入,基因转移技术已广泛用于骨组织工程领域,通过导入标志基因或治疗性基因,可致外源性调节基因表达于修复区,实现诱导成骨、免疫调节或成骨调控等预定目的。# d' W0 ~8 \! B# _
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Lou[16]以腺病毒介导hBMP2基因转移,使同质BMSCs分化为成骨细胞表型,表达了ALP活性。将其注入裸鼠体内,有异位钙化产生,且靶细胞的成骨活性依赖于腺病毒载体所表达的BMP剂量。有文献报道用腺病毒介导的TGFB1基因转染成骨细胞7 d后,靶细胞TGFB1的产生量是对照组的46倍,Ⅰ型胶原合成上升5倍,表现出良好的成骨活性[17]。金丹等[18]人使用含hBMP7基因的重组逆转录病毒感染兔BMSCs。结果显示转染后的BMSCs与对照组(空载体转染的BMSCs)在细胞增殖和细胞周期方面无显著差异,但合成胶原量上发生改变,在细胞增殖周期并无变化,所以可应用与进一步的体外构建[19.20]。相应的实验结果表明,转基因后的BMSCs在治疗骨缺损上明显优于单纯BMSCs或BMSCs复合成骨诱导剂的植入。8 Y6 U' g+ n4 m& B, V
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4问题及展望 . P+ `# r& I y' t k; F3 M2 o% K# K' D2 o( s, K
总之,关于BMSCs的研究有了较大发展,但将BMSCs应用于临床,仍面临许多问题:a)BMSCs体外定向分化培养尚未探索出成熟的方法,分离纯化细胞群的技术有待进一步提高;b)BMSCs实验仅限于自体移植方面,在异体移植免疫反应问题上有待进一步探索;c)如何促进BMSCs与载体材料相容生长并成骨的难题有待进一步解决;d)基因转染时如何有效控制调控转入基因,使之适时适量表达,同时避免不良后果。随着三维细胞培养移植模型的进一步发展、基因转染技术的完善和生物可降解材料的日趋成熟,BMSCs移植将有更广阔的前景,骨缺损、骨不连的治疗将进入一个工程化修复重建的崭新领域。1 x% E5 }* \/ G- z1 g3 v- B2 W4 C
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[20]柴岗,张艳,王敏,等.增强型绿色荧光蛋白(EGFP)转染对人骨髓基质细胞体外成骨诱导的影响[J].中华创伤骨科杂志,2003,5(2):142145.作者: kikiaaron 时间: 2010-3-23 15:08