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大鼠颅骨临界尺寸缺损用多孔纳米羟基磷灰石/聚酰胺6复合材料复合骨髓间充质干细胞修复的研究

[Reconstruction of critical sized calvarial defects by porous nano-hydroxyapatite/polyamide 6 composite with bone marrow mesenchymal stem cells in rat].

作者信息

Gao Ying, Li Ji-Hua, Li Yu-Bao, Zuo Yi, Hu Jing, Ma Yong-Qing, Wang Xue-Mei

机构信息

State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China.

出版信息

Hua Xi Kou Qiang Yi Xue Za Zhi. 2010 Feb;28(1):17-20, 24.

Abstract

OBJECTIVE

To evaluate the effects of the nano-hydroxyapatite/polyamide 6 (n-HA/PA6) on the proliferation and osteoblastic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and the feasibility of using both for constructing tissue engineered bone in the calvarias of rats with critical sized defects.

METHODS

The third passage of BMSCs were cultured in osteoblastic medium and seeded on the scaffolds of n-HA/PA6, the proliferation of the BMSCs was tested by MTT (3-{4,5-dimethylthiazol-2yl}-2,5-diphenyl-2H-tetrazolium-bromide) on scheduled dates, and the osteoblastic differentiation of the BMSCs were measured by alkaline phosphatase (ALP) staining. Furthermore, the scaffolds with or without BMSCs in rat calvarial defects, after 4 weeks, 8 weeks, and 16 weeks have been implanted. Histology and scanning electron microscope were used to test the bone healing in the different groups.

RESULTS

The BMSCs seeded on the n-HA/PA6 grew well, the proliferation of cells was not affected by the scaffold, and the staining of ALP was also positive. At 4 week and 8 week after implantation, the n-HA/PA6 with BMSCs showed more new bone formation on the surface of scaffolds, with a better osseointegration of implant and host bone when compared with the group of n-HA/PA6 without BMSCs. However, there was no significant difference between these two groups at 16 week.

CONCLUSION

The porous n-HA/PA6 has no negative effects on the proliferation and osteoblastic differentiation of rat BMSCs, and using BMSCs as seed cells and n-HA/PA6 as scaffolds is a good choice for constructing tissue engineered bone due to the enhanced new bone formation and osseointegration.

摘要

目的

评估纳米羟基磷灰石/聚酰胺6(n-HA/PA6)对大鼠骨髓间充质干细胞(BMSCs)增殖和成骨分化的影响,以及二者用于构建大鼠颅骨临界尺寸缺损组织工程骨的可行性。

方法

将第三代BMSCs在成骨培养基中培养,并接种于n-HA/PA6支架上,定期采用MTT(3-{4,5-二甲基噻唑-2-基}-2,5-二苯基-2H-四唑溴盐)检测BMSCs的增殖情况,通过碱性磷酸酶(ALP)染色检测BMSCs的成骨分化情况。此外,将含或不含BMSCs的支架植入大鼠颅骨缺损处,于4周、8周和16周后进行取材。采用组织学和扫描电子显微镜检测不同组别的骨愈合情况。

结果

接种于n-HA/PA6上的BMSCs生长良好,细胞增殖不受支架影响,ALP染色也呈阳性。植入后4周和8周,含BMSCs的n-HA/PA6支架表面有更多新骨形成,与不含BMSCs的n-HA/PA6组相比,植入物与宿主骨的骨整合更好。然而,16周时两组之间无显著差异。

结论

多孔n-HA/PA6对大鼠BMSCs的增殖和成骨分化无负面影响,将BMSCs作为种子细胞、n-HA/PA6作为支架用于构建组织工程骨是一个不错的选择,因为其能促进新骨形成和骨整合。

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