几丁糖/海藻酸敷料对海水浸泡创面作用的实验研究

EXPERIMENTAL STUDY OF THE EFFECT OF CHITOSAN/ALGINATE DRESSINGS ON WOUND IMMERSED IN SEAWATER

目的观察几丁糖(chitosan,CTS)/海藻酸(alginate,ALG)敷料对大鼠海水浸泡创面的作用。方法取健康SD大鼠25只,体重250～300g,于脊柱两侧切取直径1.8cm圆形创面,制作皮肤创面模型,左侧为实验组,右侧为对照组。将创面浸泡于预先配置的人工海水1h后,实验组采用CTS/ALG敷料外敷创面,对照组用无菌纱布外敷。行大体观察并记录创面愈合时间;术后3、5、7、10、12d切取包括创面在内的2cm×2cm皮肤标本,行HE染色和免疫组织化学Envision法染色,观察创面组织学改变及EGF受体(EGF receptor,EGFR)、bFGF表达情况。结果实验组创面炎性反应轻,结痂收缩较快;对照组创面结痂收缩较慢。实验组创面完全愈合时间为(11.68±0.57)d,对照组为(12.51±0.54)d,比较差异有统计学意义(P<0.05)。实验组自术后3d开始出现肉芽组织增生及细胞浸润,胶原组织增生,创面皱缩及上皮化,至术后10d胶原组织已排列整齐,上皮化过程基本完成,可见皮肤附件出现,术后12d完全愈合;对照组相应时间点细胞浸润出现晚,且出现肉芽组织伴溃疡,上皮化过程较晚。免疫组织化学观察示实验组术后3、5dbFGF在血管内皮细胞和间质纤维母细胞内高表达,EGFR在血管内皮细胞内高表达,术后7、10、12d呈低表达;对照组术后相应时间点bFGF及EGFR呈低表达和无表达。结论CTS/ALG对海水浸泡创面具有促进愈合作用,但其机制仍有待进一步深入研究。

Objective To observe the effect of chitosan/alginate （CTS/ALG） dressings on wound immersed in seawater. Methods Twenty-five healthy SD rats weighing 250-300g were used to establish skin wound model through cutting 1.8cm circle-shaped wound along spine bilaterally. The left side served as experimental group, and the right side as control group. The wounds were immersed in the prepared artificial seawater for 1 hour, then the experimental group was treated with CTS/ALG dressings, while the control group was treated with sterile gauze. Gross observation was performed and wound healing time was recorded. At 3, 5, 7, 10 and 12 days after operation, 2cm×2cm skin tissues including the wounds were removed and underwent HE staining and immunohistochemistry staining using Envision method. Histological change of wound and expression of EGF receptor （EGFR） and bFGF were observed. Results In the experimental group, wound inflammatory response was slight and incrustation shrinked faster, while the incrustation in the control group shrinked slowly. The wound healing time of the experimental group and the control group was （11.68±0.57） and （12.51±0.54） days, respectively, suggesting there was a significant difference between two groups （P 〈 0.05）. In the experimental group, granulation tissue proliferation, cell infiltration, collagen tissue proliferation, wound shrinkage and epithelization appeared at 3 days after operation; regularly lined collagen tissue, complete epithelization and occurrence of skin appendages were observed at 10 days after operation; complete wound healing was noted at 12 days after operation; while in the control group, at the corresponding time point, late cell infiltration and epithelization were observed and granulation tissue with ulcer was noted. Immunohistochemistry observation： high expression of bFGF in vascular endothelial cells and interstitial fibroblasts and high expression of EGFR in vascular endothelial cells were observed in the experimental group at 3 and 5 days after operation, and their expressions were low at 7, 10 and 12 days after operation; while in the control group, there were no or low expression of bFGF and EGFR at the same time point. Conclusion CTS/ALG dressings can promote the healing of wound immersed in seawater, but its mechanism needs further study.