负载真皮脱细胞基质的聚乳酸静电纺丝纤维敷料制备及检测  

Preparation and detection of acellular dermal matrix-loaded polylactic acid electrospun fiber dressings

作　　者：韩璞 李钢[1] 简玉洛[1] 孟宝玺[1] 马富廉[1] 王璐[1] Han Pu;Li Gang;Meng Baoxi;Jian Yuluo;Ma Fulian;Wang Lu(Department of Plastic Surgery,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]郑州大学第二附属医院整形外科,郑州450014

出　　处：《中华实验外科杂志》2024年第5期945-950,共6页Chinese Journal of Experimental Surgery

基　　金：河南省科技厅科技攻关项目(212102310141)。

摘　　要：目的:聚乳酸(PLA)静电纺丝纤维与真皮脱细胞基质(ADM)结合制备适宜伤口愈合的敷料。方法:利用静电纺丝工艺制备不同质量浓度(10%、12%、14%)的聚乳酸(PLA)纤维膜,筛选出最佳浓度后测定一系列生物学性能。采用4%过氧化氢-6%氢氧化钠溶液浸泡法制备兔ADM,评价脱细胞程度。将纤维膜在质量浓度为5%的ADM溶液中孵育24 h,经干燥、辐照灭菌制成负载ADM的PLA纤维膜,评价生物相容性。动物实验:在新西兰兔背部皮肤制造急性创面研究纤维膜促进创面愈合的效果。结果:12%浓度PLA纤维膜性能最佳,平均孔隙率79.37%,纤维直径分布主要范围为0.4~1.0μm,平均吸水率为550.484%,最大拉伸强度可达5.74 MPa,完全培养基孵育12周失重率为12.47%。制备的ADM无细胞残留,DNA残留量(47.528±0.419)ng/mg,与未脱细胞真皮比较,DNA去除率高达96.01%,脱细胞效果良好,差异有统计学意义(t=47.750,P<0.01)。细胞计数试剂盒(CCK-8)法检测脂肪干细胞增殖能力,48 h复合纤维膜浸提液培养吸光度值(1.121±0.013)与完全培养基培养吸光度值(0.934±0.043)比较,差异有统计学意义(t=7.181,P<0.01),说明复合纤维膜生物相容性良好。且动物实验复合纤维膜覆盖创面收缩速度最快,14 d时复合纤维膜组相对伤口面积分别与PLA组、ADM组、纱布对照组比较,差异有统计学意义(t=5.908、15.540、23.640,P<0.01)。结论:本研究制备负载ADM的PLA纤维膜性能良好,可促进兔表皮急性创面修复。Objective To prepare suitable wound healing dressings by combining polylactic acid(PLA)electrospun fibers with acellular dermal matrix(ADM).Methods The PLA fiber membranes with different mass concentrations(10%,12%,14%)were prepared by electrospinning process,and a series of biological properties were measured after screening the best concentration.The rabbit ADM was prepared by immersion method of 4%hydrogen peroxide-6%sodium hydroxide solution to evaluate the degree of decellularization.ADM-loaded PLA fibrous membrane was prepared by incubation in 5%ADM solution for 24 h,drying,irradiation sterilization,and then the biocompatibility of the fibrous membrane was evaluated.Acute wound was made on the back skin of New Zealand rabbits to study the effect of fibrous membrane on wound healing.Results 12%PLA fiber membrane had the best performance,with an average porosity of 79.37%,a fiber diameter distribution of 0.4-1.0μm,an average water absorption of 550.484%,and a maximum tensile strength of 5.74 MPa.After 12 weeks of incubation in complete medium,the weight loss rate was 12.47%.The prepared ADM had no cell residue,and the amount of DNA residue was(47.528±0.419)ng/mg.Compared with the non-acellular dermis,the DNA removal rate was as high as 96.01%,and the acellular effect was good,and the difference was statistically significant(t=47.750,P<0.01).The proliferation ability of adipose derived stem cells(ADSCs)was detected by cell counting kit-8(CCK-8)assay.The absorbance value of the composite fiber membrane extract cultured for 48 h was(1.121±0.013),which was significantly different from that of the complete medium(0.934±0.043,t=7.181,P<0.01),indicating that the composite fiber membrane had good biocompatibility.In the animal experiment,the wound covered by composite fibrous membrane contracted faster than other groups,and basically healed at 14 days.Conclusion The ADM-loaded PLA fibrous membrane prepared in this study has good performance and can promote the repair of acute epidermal wound in rabbits.

关 键 词：静电纺丝 聚乳酸 真皮脱细胞基质 纳米纤维 皮肤敷料 创面修复 

分 类 号：R622[医药卫生—整形外科]