[负载人羊膜间充质干细胞的人羊膜对SD大鼠皮肤伤口的影响]

[Effect of the human amniotic membrane loaded with human amniotic mesenchymal stem cells on the skin wounds of SD rats].

作者信息

Huo Shuang-Zhi, Shi Ping, Pang Xi-Ning

机构信息

Key Laboratory of Cell Biology of Ministry of Health, Department of Stem Cells and Regenerative Medicine, First Affiliated Hospital, China Medical University, Shenyang 110001, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2011 Dec;33(6):611-4.

PMID:22509541

Abstract

OBJECTIVE

To observe the effect of the human amniotic membrane (HAM) loaded with human amniotic mesenchymal stem cells (hAMSCs) on the skin wounds of SD rats.

METHODS

The amniotic epithelial cells were removed by trypsin digestion, hAMSCs were loaded onto HAM and then covered on rats' skin defects. The wound healing was observed by HE staining and immunohistochemistry, and the results were compared with the amniotic membrane group and blank control group.

RESULTS

The average wound healing time was (18.3 +/- 0.9) d in the HAM load with hAMSCs group, which was significantly faster than those in the blank control group [(26.4 +/- 0.7) d, P < 0.01] and the amniotic membrane group [(21.5 +/- 1.2) d, P < 0.05]. After 11 d and 14 d, the wound healing rates in the HAM load with hAMSCs group were (81.5 +/- 7.2)% and (94.3 +/- 3.6)%, respectively, which were significantly higher than those in the blank control group [(48.5 +/- 3.2)% and (74.3 +/- 4.3 )%] and the amniotic membrane group [(68.5 +/- 4.5)% and (86.8 +/- 4.8)%] (all P < 0.01). Skin biopsy/HE staining confirmed that the quality of wound healing in the HAM load with hAMSCs group was significantly better than in the amniotic membrane group and the blank control group. Immunohistochemical staining showed that the number of CK19-positive epidermal stem cells in the HAM load with hAMSCs group (48.2 +/- 3.2) was significantly larger than those in the amniotic membrane group (37.7 +/- 3.1) (P < 0.05) and the blank control group (29.6 +/- 2.4) (P < 0.01). Furthermore, the vascular endothelial growth factor expression (64.5 +/- 4.5) in the HAM load with hAMSCs group was also significantly higher than those in the amniotic membrane group (52.6 +/- 3.8) (P < 0.05) and the blank control group (40.7 +/- 3.1) (P < 0.01).

CONCLUSION

HAM loaded with hAMSCs may promote the repair of skin wounds by promoting the regeneration of epidermal stem cells and capillaries.

摘要

目的

观察负载人羊膜间充质干细胞（hAMSCs）的人羊膜（HAM）对SD大鼠皮肤伤口的影响。

方法

用胰蛋白酶消化去除羊膜上皮细胞，将hAMSCs接种到HAM上，然后覆盖在大鼠皮肤缺损处。通过苏木精-伊红（HE）染色和免疫组织化学观察伤口愈合情况，并将结果与羊膜组和空白对照组进行比较。

结果

负载hAMSCs的HAM组平均伤口愈合时间为（18.3±0.9）天，明显快于空白对照组[（26.4±0.7）天，P<0.01]和羊膜组[（21.5±1.2）天，P<0.05]。11天和14天后，负载hAMSCs的HAM组伤口愈合率分别为（81.5±7.2）%和（94.3±3.6）%，明显高于空白对照组[（48.5±3.2）%和（74.3±4.3）%]和羊膜组[（68.5±4.5）%和（86.8±4.8）%]（均P<0.01）。皮肤活检/HE染色证实，负载hAMSCs的HAM组伤口愈合质量明显优于羊膜组和空白对照组。免疫组织化学染色显示，负载hAMSCs的HAM组细胞角蛋白19（CK19）阳性表皮干细胞数量（48.2±3.2）明显多于羊膜组（37.7±3.1）（P<0.05）和空白对照组（29.6±2.4）（P<0.01）。此外，负载hAMSCs的HAM组血管内皮生长因子表达（64.5±4.5）也明显高于羊膜组（52.6±3.8）（P<0.05）和空白对照组（40.7±3.1）（P<0.01）。