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目的研究鲨鱼皮胶原(Shark Skin Collagen,SSC)海绵的体外抗氧化功效及其体内促伤口愈合作用。方法采用醋酸溶胀法分离鲨鱼皮胶原,真空冷冻干燥技术制备SSC海绵,化学分光光度法检测其体外抗氧化活性;采用SD大鼠背部创伤模型,检测SSC海绵的促愈合作用,HE染色观察组织病理学变化,免疫组织化学染色法检测愈合过程中CD31,EGF和TGF-β表达变化。结果 20mg/mL SSC溶液对HO·清除率为(84±2.1)%。术后第3、5天,SSC海绵组愈合率分别为(52.1±5.7)%、(57.1±8.0)%,显著优于纱布(gauze,GZ)组(15.9±8.3)%、(28.4±6.6)%(P<0.01)和壳聚糖(chitosan,CS)组(22.2±9.5)%、(36.5±3.3)%(P<0.05)。HE染色显示,SSC海绵能在创伤愈合早期显著促进毛细血管、新生肉芽组织和愈合上皮生成,加速创面愈合。免疫组化检测显示,术后第3天,SSC海绵组CD31管状表达数为38±4.73,显著高于CS组27±3.06和GZ组26±3.61(P<0.05);SSC海绵组TGF-β表达率为(20.1±3.17)%,显著高于GZ组(6.7±0.739)%和CS组(10.7±0.502)%(P<0.05);SSC海绵组EGF表达率为(12.1±0.447)%,显著高于GZ组(9.3±1.23)%(P<0.05)。结论 SSC具有显著的清除羟自由基功效,SSC海绵能够显著促进创伤愈合早期CD31、TGF-β和EGF的高表达,进而促进早期新血管生成和肉芽组织的形成,加快伤口愈合过程。
Objective To study the anti-oxidative effect of Shark Skin Collagen (SCC) sponge in vitro and its healing effect on wound healing. METHODS: Shark skin collagen was isolated by acetic acid swelling method and SSC sponge was prepared by vacuum freeze-drying technique. The anti-oxidative activity of SSC in vitro was detected by chemical spectrophotometry. The healing of SSC sponge was detected by the model of trauma to the back of SD rats. Neutrophil histological changes and immunohistochemical staining were used to detect the expression of CD31, EGF and TGF-β in the healing process. Results The clearance rate of HO · in 20mg / mL SSC solution was (84 ± 2.1)%. The healing rates of SSC sponge group were (52.1 ± 5.7)% and (57.1 ± 8.0)% on the 3rd and 5th day postoperatively respectively, which were significantly better than those of the gauze group (15.9 ± 8.3)% and (28.4 ± 6.6) ) (P <0.01) and chitosan group (22.2 ± 9.5)% and (36.5 ± 3.3)% respectively (P <0.05). HE staining showed that SSC sponge could significantly promote the formation of capillaries, nascent granulation tissue and healing epithelium in the early stage of wound healing and accelerate wound healing. Immunohistochemistry showed that the number of CD31 in SSC sponge group was 38 ± 4.73 on the 3rd postoperative day, which was significantly higher than that in CS group (27 ± 3.06) and GZ group (26 ± 3.61) (P <0.05) The expression rate of EGF in SSC sponge group was (20.1 ± 3.17)%, which was significantly higher than that in GZ group (6.7 ± 0.739)% and CS group (10.7 ± 0.502)% Which was significantly higher than that of GZ group (9.3 ± 1.23)% (P <0.05). Conclusion SSC has a significant effect of scavenging hydroxyl radical. SSC sponge can significantly promote the high expression of CD31, TGF-β and EGF in the early stage of wound healing, and then promote the formation of early neovascularization and granulation tissue and accelerate the wound healing process.