Healing of critically sized femoral defects, using genetically modified mesenchymal stem cells from human adipose tissue
Healing of critically sized femoral defects, using genetically modified mesenchymal stem cells from human adipose tissue
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DOI：
10.1089/ten.2005.11.120
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发表时间：
2005-01-01
影响因子：
--
通讯作者：
Lieberman, JR
中科院分区：
文献类型：
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作者：
Peterson, B;Zhang, J;Lieberman, JR
FDA 已批准重组骨形态发生蛋白 (BMP) 的临床使用。然而，在人体中使用重组 BMP 需要大剂量的蛋白质才能有效，这表明骨形态发生蛋白的递送方法需要优化。基因疗法是传递此类重组蛋白的另一种方法，基因转移技术已在多种细胞类型上进行了测试，包括骨髓细胞、皮肤成纤维细胞、外周血单核细胞和肌肉来源的细胞。在这项研究中，我们试图确定产生 BMP-2 的人脂肪来源间充质干细胞治愈裸鼠模型中严重股骨缺损的能力。经人类受试者保护委员会批准后，从健康捐赠者身上获取人体脂肪组织。如前所述处理脂肪抽吸物（De Ugarte, D. A., et al. Cells Tissues Organs 174, 101, 2003）。细胞在培养物中生长并用携带 BMP-2 的腺病毒感染。将五百万个细胞应用至胶原陶瓷载体并植入股骨缺损中，如先前所述(Zuk，P.A.，等人，Mol.Biol.13, 4279, 2002)。所有动物在第8周时被处死。解剖股骨并进行放射线照相、组织学和生物力学分析。在用经过基因修饰以过表达 BMP-2 的人加工脂肪抽吸物 (HPLA) 细胞治疗的组中，12 个股骨中的 11 个在第 8 周时已愈合。这是通过射线照片、机械测试和组织学来评估的。未愈合的一根股骨患有亚急性感染。用rhBMP-2-浸渍的胶原蛋白-陶瓷载体治疗的所有八个股骨均愈合。这两组之间没有检测到统计学上的显着差异。对照组的评估：第II组（胶原蛋白-陶瓷载体与HPLA细胞）和第III组（仅胶原蛋白-陶瓷载体）显示8周时没有股骨愈合。我们的结果表明，通过腺病毒基因转移进行基因修饰以过表达 BMP-2 的 HPLA 细胞可以诱导体内骨形成，并治愈无胸腺大鼠的严重股骨缺损。单独的 HPLA 细胞不会诱导显着的骨形成。然而，当与骨诱导因子结合时，这些细胞可能是增强骨愈合和骨组织工程的有效方法。
The FDA has approved the clinical use of recombinant bone morphogenetic proteins (BMPs). However, the use of recombinant BMPs in humans has required large doses of the proteins to be effective, which suggests that the delivery method of bone morphogenetic proteins needs to be optimized. Gene therapy is an alternative method to deliver such recombinant proteins, and gene transfer techniques have been tested on a variety of cell types including bone marrow cells, skin fibroblasts, peripheral blood monocytes, and muscle-derived cells. In this study, we sought to determine the ability of BMP-2-producing human adipose-derived mesenchymal stem cells to heal a critically sized femoral defect in a nude rat model. After approval by the human subjects protection committee, human adipose tissue was obtained from healthy donors. The lipoaspirate was processed as previously described ( De Ugarte, D. A., et al. Cells Tissues Organs 174, 101, 2003). Cells were grown in culture and infected with a BMP-2-carrying adenovirus. Five million cells were applied to a collagen -ceramic carrier and implanted into femoral defects as previously described (Zuk, P. A., et al. Mol. Biol. 13, 4279, 2002). All animals were killed at 8 weeks. Femora were dissected out and underwent radiographic, histologic, and biomechanical analysis. Eleven of the 12 femora in the group treated with human processed lipoaspirate ( HPLA) cells genetically modified to overexpress BMP- 2 had healed at 8 weeks. This was assessed by radiographs, by mechanical testing, and by histology. The one femur that did not heal had a subacute infection. All eight of the femora treated with the rhBMP-2- impregnated collagen - ceramic carrier healed. No statistically significant difference was detected between these two groups. Evaluation of the control groups: group II ( collagen - ceramic carrier with HPLA cells) and group III ( collagen - ceramic carrier alone) showed that none of the femora had healed by 8 weeks. Our results indicate that HPLA cells genetically modified by adenoviral gene transfer to overexpress BMP- 2 can induce bone formation in vivo and heal a critically sized femoral defect in an athymic rat. The HPLA cells alone did not induce significant bone formation. However, when combined with an osteoinductive factor these cells may be an effective method for enhancing bone healing and the tissue engineering of bone.