新型静电纺丝伤口敷料的止血性能及促愈合作用研究

Study on hemostatic property and wound healing effects of new electrostatic spinning wound dressing

目的评价新型静电纺丝伤口敷料的止血性能及促创伤愈合作用,为该敷料的进一步开发应用奠定基础。方法将胶原蛋白和壳聚糖的混合溶液通过静电纺丝技术纺织在海藻酸盐无纺布基布上,制备成新型静电纺丝伤口敷料(胶原蛋白-壳聚糖静电纺丝膜复合海藻酸盐敷料CCEAD)。通过体外凝血实验、兔耳动脉创伤止血实验和兔背部创伤止血实验评价CCEAD的止血性能。建立大鼠背部全皮层创伤模型,分别在术后3、5、7、12d拍照计算伤口愈合率,测定CCEAD的促愈合作用;取伤口创面皮肤组织,HE染色观察组织病理学变化;免疫组化染色观察在伤口愈合过程中CD31、FGF-2和TGF-β的表达情况。结果 CCEAD组的体外凝血指数BCI为36.91%±6.62%,显著小于阳性对照明胶海绵组(69.82%±1.83%)和阴性对照医用纱布组(79.93%±6.30%)(P<0.01)。兔耳动脉创伤止血实验中,CCEAD组的止血时间(107.67±17.62)s短于医用纱布组(163.67±15.50)s(P<0.05)。兔背部创伤止血实验中,CCEAD组Hb光度吸收值(0.120±0.052)明显低于医用纱布组(0.294±0.021)(P<0.01)。在促创伤愈合实验中,术后第7天,CCEAD组愈合率为65.70%±4.94%,高于阳性对照壳聚糖敷料组(48.78%±8.33%)(P<0.05),明显高于医用纱布组(37.76%±6.35%)(P<0.01)。HE染色显示,CCEAD在伤口愈合早期能够促进成纤维细胞生长和新生血管形成,促进肉芽组织的生成。免疫组化结果显示,CCEAD能促进CD31、FGF-2和TGF-β在伤口愈合早期的表达,有利于伤口的愈合。结论新型静电纺丝伤口敷料(胶原蛋白-壳聚糖静电纺丝膜复合海藻酸盐敷料)止血性能优良,具有明显的促创伤愈合作用,有望开发成新型伤口敷料。

Objective To evaluate the hemostatic property and wound healing effects of new electrostatic spinning wound dressing,and lay a foundation for its further development and application.Methods The new electrostatic spinning wound dressing(collagen-chitosan electrostatic spinning membrane alginate dressing,CCEAD)was prepared by spinning the mixture of collagen and chitosan onto alginate non-woven fabric substrate by electrospinning technology.The hemostatic properties of CCEAD were evaluated by Blood Clotting Index(BCI)test,hemostasis test of rabbit ear artery trauma and hemostasis test of rabbit back trauma.Rat back wound model was used to evaluate the wound healing effects.The wound healing rate was calculated by taking pictures on the 3rd,5th,7th and 12th day after operation.HE staining was used to observe histopathological changes.Immunohistochemical staining was used to observe expression of CD31,FGF-2 and TGF-β.Results The BCI of CCEAD group was 36.91%±6.62%,which was significantly less than gelatin sponge group(69.82%±1.83%)and gauze group(79.93%±6.30%)(P<0.01).In the hemostasis test of rabbit ear artery trauma,the hemostatic time of CCEAD group(107.67±17.62)s was significantly shorter than that of gauze group(163.67±15.50)s(P<0.05).In the hemostasis test of rabbit back trauma,the absorbance value of Hb in CCEAD group(0.120±0.052)was significantly lower than that of gauze group(0.294±0.021)(P<0.01).In the wound healing test,on the 7th day after operation,the healing rate of CCEAD group was 65.70%±4.94%,which was higher than that of chitosan dressing group(48.78%±8.33%)(P<0.05)and significantly higher than gauze group(37.76%±6.35%)(P<0.01).HE staining showed that CCEAD could promote fibroblast growth,angiogenesis and granulation tissue formation in the early stage of wound healing.Immunohistochemical results showed that CCEAD could promote the expression of CD31,FGF-2 and TGF-βin the early stage of wound healing,which was beneficial to wound healing.Conclusion The new electrostatic spinning wound dressing(collagen-chitosan electrostatic spinning membrane alginate dressing)has excellent hemostatic properties and obvious effect of promoting wound healing.It is expected to be developed into a new wound dressing.