Influences of vacuum assisted closure on the expression and distribution of
metalloproteinases in human chronic wound☆

Abstract

AIM：To observe the effects of vacuum assisted closure (VAC) on expressions of metalloproteinases (MMPs) MMP-1，MMP-2，MMP-8，MMP-9，MMP-13 and tissue inhibitor of metalloproteinases (TIMPs) TIMP-1，TIMP-2 in margin tissues of human chronic wounds.

METHODS： ①Nine patients with chronic wounds were selected from the Tangdu Hospital, Fourth Military Medical University of Chinese PLA from December 2001 to January 2003. The patients were with wound forming for 1 month, but without healing trend, and were treated with traditional method, but inefficiency. All the patients were treated with VAC. Hole of medical sponge was 400－600 μm and adjusted negative pressure was －15.96 kPa for successive suction, and dressing was changed 1, 4 and 7 days after sucking. ②Immunohistochemical method was used to observe the expressions and distributions of MMP-1，MMP-2，MMP-8，MMP-9，MMP-13，TIMP-1，TIMP-2 protein in margin tissues 1, 4, 7 days before and after treatment with VAC, respectively, and then dynamic comparisons were performed.

RESULT: Totally 9 patients were involved in the results analysis. In general, expressions of MMP-1，MMP-2，MMP-8，MMP-9，MMP-13 became less, but expressions of TIMP-1 and TIMP-2 became strong with time going after VAC. More MMP-1 expressions were found in keratinocyte layer with no obvious changes in dermis after VAC. Not only could we observed the expressions of MMP-8 around inflammatory cells, but also could observed the expressions in epidermis with no changes before and after VAC. But less expression could be found in dermis after VAC. Before VAC, MMP-13 stain could be found in keratinocyte layer, but disappeared after VAC. Mainly changes were that stain became less in dermis after VAC with becoming less as approaching the wound margin. There was less stain in keratinocyte layer and more stain in basal layer in MMP-2, and distinctly more stain in epidermis in MMP-9 at day 7 after VAC. Before VAC no TIMP-1 and TIMP-2 expressions could be found, but expressions became stronger after VAC in epidermis and dermis, and was the deepest in the granular cell layer as well as stain being deep around blood vessels.

CONCLUSION: ①Hydrolysis of collagen protein is depressed through VAC inhibiting MMP expression and promoting TIMP expression. ②MMP-1，MMP-2，MMP-9 might play role in improving the keratinocyte immigrating and MMP-8 and MMP-13 might be involved in granulation tissues remodeling after VAC. ③TIMP-1 and TIMP-2 might be secreted by epidermis, and after VAC TIMP-1 and TIMP-12 not only depress the MMP expression but also provide a protective role for blood vessels.

Shi B, Qian CR, Kuang F, Li JQ, Chen SZ, Li WZ, Li XY.Influences of vacuum assisted closure on the expression and distribution of metalloproteinases in human chronic wound.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2007;11(13):2482-2486(China)
[www.zglckf.com/zglckf/ejournal/upfiles/07-13/13k-2482(ps).pdf]

摘要
目的：观察封闭负压引流术对人慢性创面创缘组织中金属基质蛋白酶（metalloproteinases，MMPs）MMP-1，MMP-2，MMP-8，MMP-9，MMP-13以及其组织抑制剂（tissue inhibitor of metalloproteinases，TIMPs）TIMP-1，TIMP-2蛋白的影响。
方法：①选取2001－12/2003－01解放军第四军医大学唐都医院收治的慢性创面（创面形成1个月以上而无愈合倾向，应用保守治疗无效）患者9例，应用封闭负压引流术治疗，医用海绵孔径400～600 μm，调整负压为－15.96 kPa，连续吸引，在吸引后1，4，7 d更换敷料。②分别于治疗前和治疗后1，4，7 d切取创面创缘组织，用免疫组织化学方法观察MMP-1，MMP-2，MMP-8，MMP-9，MMP-13，TIMP-1，TIMP-2的蛋白表达及分布，并进行动态比较。
结果：9例均进入结果分析。总体上随封闭负压引流作用时间延长，MMP-1，MMP-2，MMP-8，MMP-9，MMP-13的含量降低，而TIMP-1和TIMP-2水平增高。封闭负压引流后MMP-1在表皮角化层含量增加，真皮层变化不很显著；在炎细胞周围和表皮中均发现MMP-8染色，治疗前后变化不很明显，真皮染色渐淡；治疗前表皮角化层有MMP-13的染色，治疗后不仅表皮染色消失，而且主要是真皮弥漫性分布变淡，越近创缘染色越浅；MMP-2在角化层染色变淡，基底层染色加深；MMP-9则表皮在7 d时染色加深明显；TIMP-1，TIMP-2在治疗前基本无染色，治疗后表皮真皮均出现染色，最深在颗粒细胞层，血管壁周围染色加深。
结论：①封闭负压引流通过抑制MMP，促进TIMP分泌使胶原蛋白水解受到抑制。②MMP-1，MMP-2，MMP-9共同在封闭负压引流促进表皮迁移中起作用，MMP-8与MMP-13则主要是参与了封闭负压引流后的肉芽组织重塑。③TIMP-1和TIMP-2主要由表皮细胞分泌，封闭负压引流后不仅抑制了MMP的作用，更重要的是对血管的保护作用。
关键词：封闭负压引流；金属基质蛋白酶；免疫组化；伤口愈合；慢性病

石冰，钱存荣，邝芳，李金清，陈绍宗，李望舟，李学拥.封闭负压引流技术对人慢性创面中金属基质蛋白酶表达及分布的影响[J].中国组织工程研究与临床康复，2007，11(13):2482-2486 [www.zglckf.com/zglckf/ejournal/upfiles/07-13/13k-2482(ps).pdf]