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[胶原蛋白-壳聚糖真皮支架修复全层皮肤缺损中生物合成与细胞凋亡的机制及作用]

[Mechanisms and effects of biosynthesis and apoptosis in repair of full-thickness skin defect with collagen-chitosan dermal stent].

作者信息

Xu Shao-Jun, Huang Ai-bin, Ma Lie, Teng Jian-Ying, Gao Chang-You, Zhang Zhi-Liang, Ni You-Di, Ye Sheng, Wang Yong-Guang

机构信息

Department of Plastic Surgery of the Affiliated Hospital of Hangzhou Normal University, Hangzhou 310015, China.

出版信息

Zhonghua Zheng Xing Wai Ke Za Zhi. 2009 May;25(3):208-12.

Abstract

OBJECTIVE

To investigate biosynthetic and apoptotic mechanisms in repair of full thickness skin defect with collagen-chitosan porous scaffold transplantation, and to determinate differences between wound repair with the scaffold transplantation and scar healing without the scaffold transplantation.

METHODS

The full thickness skin defects were made on 10 Bama miniature pigs and the bilayer dermal equivalent (BDE) composed of collagen-chitosan porous scaffold and silicone membrane was transplanted on wounds. Surfaces of wounds were observed at 1, 2, and 3 weeks after the BDE transplantation, and so were done the wound repairs after epidermis had been grafted for 2 weeks on surface of the scaffold which had been transplanted on skin defect wounds for 2 weeks. At the same time, TGF-beta1 expressions, apoptosis and self collagen replacement of scaffolds in wounds were detected in situ by immunohistochemical staining, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) and picrosirius red polarized light. Wounds without scaffold transplantation were studied as control.

RESULTS

  1. Wounds with the scaffold transplantation were different from granulation tissue. 2) The peak of TGF-beta1 expression in the scaffold wounds was from 1 to 2 weeks after BDE transplantation, and TGF-beta1 expressions decreased continuously from 3 to 4 weeks. TGF-beta1 expressions increased continuously in the control wounds from 1 to 3 weeks and decreased on 4 weeks. TGF-beta1 expressions in the scaffold wounds on 1st and 2nd week were significantly higher than those in the corresponding control wounds, whereas, TGF-beta1 expressions in the scaffold wounds on 3rd and 4th week were significantly lower than those in the corresponding control wounds. 3) Apoptosis increased continuously in the scaffold wounds from 2 to 4 weeks after BDE transplantation, and so did in the control wounds from 3 to 4 weeks. However, apoptosis signals in the scaffold wounds on 2nd, 3rd, and 4th week after BDE transplantation were significantly more than those in the corresponding control wounds, and there was no difference between apoptosis signals in the scaffold wounds on 1st week after BDE transplantation and those in the corresponding control wounds. 4) Observation by picrosirius red polarized light method: self collagen began to synthesize in the scaffold wounds on 1st week after BDE transplantation, and scaffolds had been replaced by self collagen from 2 to 3 weeks after BDE transplantation.

CONCLUSIONS

Collagen-chitosan porous scaffold plays a very important role in wound healing of full thickness skin defect. The mechanisms of wound repair by dermal scaffold are different from those by granulation and scar healing. It has a good future in repairing skin defect.

摘要

目的

探讨胶原 - 壳聚糖多孔支架移植修复全层皮肤缺损的生物合成及凋亡机制,并确定支架移植伤口修复与无支架移植瘢痕愈合之间的差异。

方法

在10只巴马小型猪身上制作全层皮肤缺损,将由胶原 - 壳聚糖多孔支架和硅酮膜组成的双层真皮替代物(BDE)移植到伤口上。在BDE移植后1、2和3周观察伤口表面情况,在皮肤缺损伤口上移植支架2周后,再在支架表面移植表皮2周后观察伤口修复情况。同时,通过免疫组织化学染色、末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸 - 生物素缺口末端标记法(TUNEL)和苦味酸天狼星红偏振光原位检测伤口中支架的转化生长因子 - β1(TGF - β1)表达、凋亡及自身胶原替代情况。以未进行支架移植的伤口作为对照。

结果

1)支架移植的伤口与肉芽组织不同。2)支架伤口中TGF - β1表达的峰值出现在BDE移植后1至2周,3至4周TGF - β1表达持续下降。对照伤口中TGF - β1表达在1至3周持续上升,4周时下降。支架伤口在第1周和第2周的TGF - β1表达显著高于相应对照伤口,而支架伤口在第3周和第4周的TGF - β1表达显著低于相应对照伤口。3)BDE移植后2至4周支架伤口中的凋亡持续增加,对照伤口在3至4周凋亡也增加。然而,BDE移植后第2、3和4周支架伤口中的凋亡信号明显多于相应对照伤口,BDE移植后第1周支架伤口中的凋亡信号与相应对照伤口无差异。4)苦味酸天狼星红偏振光法观察:BDE移植后第1周支架伤口开始合成自身胶原,BDE移植后2至3周支架被自身胶原替代。

结论

胶原 - 壳聚糖多孔支架在全层皮肤缺损伤口愈合中起非常重要的作用。真皮支架伤口修复机制与肉芽及瘢痕愈合不同。其在修复皮肤缺损方面具有良好前景。

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