手持静电纺丝可降解纳米纤维原位修复全层皮肤缺损  

In situ repair of full-thickness skin defects by handheld electrospun biodegradable nanofibers

作　　者：陈洪让 张海涛[3] 邓坤学 李永生 沈云[2] 徐弢[4] 章新琼[1] Chen Hongrang;Zhang Haitao;Deng Kunxue;Li Yongsheng;Shen Yun;Xu Tao;Zhang Xinqiong(Nursing College,Anhui Medical University,Hefei 230032,Anhui Province,China;Department of General Surgery,the First Affiliated Hospital of Anhui Medical University,Hefei 230032,Anhui Province,China;Guangzhou Medprin Regenerative Medicine Technology Co.,Ltd.,Guangzhou 510663,Guangdong Province,China;Department of Mechanical Engineering,Biomanufacturing Center,Tsinghua University,Beijing 100084,China)

机构地区：[1]安徽医科大学护理学院,安徽省合肥市230032 [2]安徽医科大学第一附属医院普外科,安徽省合肥市230032 [3]广州迈普再生医学科技股份有限公司,广东省广州市510663 [4]清华大学机械工程系生物制造中心,北京市100084

出　　处：《中国组织工程研究》2019年第2期257-264,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家高新技术研究与发展规划863项目(2015AA020303;项目负责人:徐弢)~~

摘　　要：背景:传统静电纺丝纳米纤维制造过程相对复杂,制造条件要求高,无法适应创伤、烧/烫伤等组织缺损的应急事件中快速组织修复的需求。目的:观察手持式静电纺丝聚乳酸/明胶可降解纳米纤维膜对小鼠皮肤缺损原位修复的效果。方法:(1)采用自制3D打印的手持式静电纺丝设备制备聚乳酸/明胶可降解纳米纤维膜,检测其接触角、水蒸气透过率;(2)以浓度100%,50%,20%的聚乳酸/明胶可降解纳米纤维膜浸提液培养胎鼠成纤维细胞,采用CCK-8细胞毒实验评估材料残留溶剂毒性;将胎鼠成纤维细胞与聚乳酸/明胶可降解纳米纤维膜共培养(实验组),设置单独细胞培养为对照,Alamar blue法检测细胞增殖,活/死染色观察细胞存活率,扫描电镜观察细胞形态;(3)在18只Balb/c小鼠背部制作直径2cm皮肤全层缺损,实验组进行手持静电纺丝原位聚乳酸/明胶可降解纳米纤维膜修复后纱布包扎,对照组进行纱布包扎,术后8周进行缺损部位苏木精-伊红和Masson染色,观察缺损皮肤修复效果。结果与结论:(1)聚乳酸/明胶可降解纳米纤维膜的接触角为(32.68±5.68)°,属亲水材料,适宜细胞黏附;24 h水蒸气透过率为(4.21±0.11)×10~3 g/m^2,可满足皮肤外敷料的要求;不同浓度的聚乳酸/明胶可降解纳米纤维膜浸提液无明显的细胞毒性;(2)实验组胎鼠成纤维细胞具有与对照组细胞等同的细胞活性,但具有更快的增殖速度与更长的增殖时间;(3)苏木精-伊红和Masson染色显示,实验组小鼠皮肤伤口全层愈合,材料降解完全,毛囊再生;对照组小鼠皮肤未全层愈合;(4)结果表明,手持式静电纺丝聚乳酸/明胶可降解纳米纤维实现小鼠皮肤全层缺损的原位修复。BACKGROUND:The traditional electrospinning nanofiber manufacturing process is relatively complicated,which requires high manufacturing conditions and cannot meet the needs of rapid tissue repair in emergency events such as trauma and burn/scald.OBJECTIVE:To observe the effect of handheld electrospun polylactic acid/gelatin degradable nanofiber membrane on the in situ repair of mouse skin defects.METHODS:Handheld electrospun polylactic acid/gelatin degradable nanofiber membranes were prepared by self-made 3D printing handheld electrospinning device,and the contact angle and water vapor transmission rate were then measured.Fetal rat fibroblasts were cultured with 100%,50%,20%polylactic acid/gelatin degradable nanofiber membrane extracts,and the residual solvent toxicity of the materials was evaluated by cell counting kit-8 cytotoxicity assay.Fetal rat fibroblasts were co-cultured with polylactic acid/gelatin degradable nanofiber membrane(experimental group),and cells cultured alone were set as control.Cell proliferation was detected by Alamar blue method,cell viability was observed by live/dead staining,and cell morphology was observed by scanning electron microscope.A full-thickness skin defect of 2 cm in diameter was made on the back of 18 Balb/c mice.The experimental group was inlaid with handheld electrospun polylactic acid/gelatin degradable nanofiber membrane for in situ repair followed by gauze dressing.The control group was treated with gauze dressing at the defect site.Eight weeks after operation,hematoxylin-eosin and Masson staining were used to observe the repair of skin defects.RESULTS AND CONCLUSION:(1)The contact angle of polylactic acid/gelatin degradable nanofiber membrane was(32.68±5.68)°,indicating a hydrophilic material suitable for cell adhesion.The 24-hour water vapor transmission rate was(4.21±0.11)×103 g/m2,which met the requirements of external skin dressing.Different concentrations of polylactic acid/gelatin degradable nanofiber membrane extract had no obvious cytotoxicity.(2)In the exp

关 键 词：可降解纳米纤维 原位修复 手持静电纺丝 聚乳酸/明胶 生物材料 纳米纤维 明胶 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]