Tissue-engineered bone formation with gene transfer and mesenchymal stem cells in a minimally invasive technique
Tissue-engineered bone formation with gene transfer and mesenchymal stem cells in a minimally invasive technique
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DOI：
10.1097/mlg.0b013e31805f680e
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发表时间：
2007-07-01
期刊：
影响因子：
2.6
通讯作者：
Eun, Hee Seog
中科院分区：
文献类型：
--
作者：
Park, Dong-Joon;Choi, Jin Hyun;Eun, Hee Seog
背景：本研究的目的是利用壳聚糖-海藻酸盐凝胶以微创方式皮下植入骨髓间充质干细胞，并通过同时转移的成骨蛋白（OP）-1（骨形态发生蛋白-7）基因促进骨形成。方法和结果：嵌入凝胶中的聚乙烯亚胺/荧光素酶质粒DNA复合物能够转染培养皿上或共封装在培养皿中的HEK 293细胞。凝胶。当注射到小鼠的皮下空间时，荧光素酶的表达比背景增加了两到三个数量级。为了检查基因、细胞以及组合的基因和细胞封装凝胶在组织生成中的功效，将样品注射到 6 周大无胸腺裸鼠的皮下空间，并对 OP-1 质粒进行研究。注射后8周，只有当人间充质干细胞（hMSC）被封装时，凝胶才保持其体积形状，否则凝胶会部分溶解。 4 周后在样品中清楚地检测到 OP-1 的转基因表达，但 8 周后未检测到。在所有含有 OP-1 质粒的凝胶中均检测到 II 型胶原蛋白，无论是否含有 hMSC。 OP-1 DNA 和 hMSC 组合的样品显示出强烈的 II 型胶原蛋白表达以及类骨病灶。结论：这些结果表明，在壳聚糖-海藻酸盐凝胶内组合基因和 hMSC 递送可能是组织工程的一种有趣的方法。
Background: The objective of this study was to use a chitosan-alginate gel to implant bone marrow-derived mesenchymal stem cells subcutaneously in a minimally invasive manner and promote bone formation by the simultaneously transferred osteogenic protein (OP)-1 (bone morphogenic protein-7) gene.Method and Results: The complex of polyethylenimine/luciferase plasmid DNA embedded in the gel was able to transfect HEK 293 cells on a culture dish or co-encapsulated in the gel. When injected into the subcutaneous space of mice, luciferase expression was two to three orders of magnitude increased above the background. To examine the efficacy of gene-, cell-, and combined gene- and cell-encapsulated gels in tissue generation, samples were injected into the subcutaneous space of 6-week-old athymic nude mice, and the OP-1 plasmid was studied. At 8 weeks after the injection, the gels only maintained their volumetric shape when human mesenchymal stem cells (hMSCs)were encapsulated, but otherwise the gels were partially dissolved. Transgene expression of OP-1 was clearly detected in the samples after 4 weeks but not after 8 weeks. Type II collagen was detected in all of the gels containing the OP-1 plasmid, with or without hMSCs. The samples with the combination of OP-1 DNA and hMSCs revealed strong type II collagen expression as well as osteoid foci.Conclusion: These results suggest that combined gene and hMSC delivery within a chitosan-alginate gel could be an interesting approach for tissue engineering.