纳米聚吡咯/甲壳素复合膜的制备及其生物相容性观察  

Preparation and biocompatibility of nano polypyrrole/chitin composite membrane

作　　者：焦海山[1] 曹萍 陈颖 宋悦宁[1] 李东印[1] 王晓冬 JIAO Haishan;CAO Ping;CHEN Ying;SONG Yuening;LI Dongyin;WANG Xiaodong(School of Clinical Medicine,Suzhou Vocational Health College,Suzhou ]iangsu,215009,P.R.China;Department of Histology and Embryology,Medical School of Nantong University,Nantong Jiangsu,226001,P.R.China)

机构地区：[1]苏州卫生职业技术学院临床医学院,江苏苏州215009 [2]南通大学医学院组织学与胚胎学系,江苏南通226001

出　　处：《中国修复重建外科杂志》2018年第8期1081-1087,共7页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：江苏省卫生厅卫生职业技术教育研究室立项课题(J201301);江苏省高等学校自然科学研究项目(18KJD310005);苏州市科技局项目(STSD2011074);江苏省"青蓝工程"资助项目(苏教师[2012]39号)~~

摘　　要：目的制备纳米聚吡咯(polypyrrole,PPy)/甲壳素复合膜并观察其生物相容性。方法采用微乳液聚合法合成纳米PPy,与壳聚糖共混并成膜后,进行乙酰化改性得到纳米PPy/甲壳素复合膜(A组),制备壳聚糖膜(B组)及经乙酰化改性得到的甲壳素膜(C组)作为对照组,使用扫描电镜、红外光谱观察等方法对纳米PPy、各组膜进行鉴定并测定其导电性。将3组膜与雪旺细胞体外共培养,采用倒置显微镜观察、活细胞染色、细胞计数、免疫荧光染色等方法观察膜的体外生物相容性,使用溶菌酶溶液评价膜的体外降解情况。结果纳米PPy合成后,经红外光谱观察,显示在1 543.4 cm^(–1)和1 458.4 cm^(–1)处出现PPy C=C的特征振动吸收峰;扫描电镜观察发现纳米PPy的粒子聚合体直径为100~200 nm。3组膜制备后,红外光谱观察显示,A、C组膜出现乙酰化改性后的1 562 cm^(–1)左右的酰胺Ⅱ谱带特征峰,显示A、C组膜成功乙酰化成甲壳素。导电性检测显示A组膜的电导率为(1.259 2±0.005 7)×10–3 S/cm;B、C组膜均未检测出电导率。扫描电镜观察到A组膜表面有均匀分布的纳米PPy聚合颗粒,对照组光滑平整,显示纳米PPy与壳聚糖共混并改性后成功得到导电纳米PPy/甲壳素复合膜。将雪旺细胞与3组膜共培养,经二乙酸荧光素活细胞染色、可溶性蛋白-100免疫荧光染色及倒置显微镜观察发现,培养的雪旺细胞存活,功能状态良好;共培养2、4 d后,细胞计数显示A组膜上细胞增殖数量显著多于B、C组(P<0.05),显示A组膜表现出比对照组更强的支持细胞黏附增殖的能力。膜的体外降解观察发现各时间点A、C组膜体外降解率均显著高于B组(P<0.05),表明同等条件下乙酰化改性处理的膜降解性能优于壳聚糖膜。结论纳米PPy与壳聚糖可成功共混并顺利乙酰化改性,所得纳米PPy/甲壳素复合膜导电可降解,具有较好的体外生物相容性。Objective To prepare nano polypyrrole（PPy）/chitin composite membrane and observe their biocompatibility. Methods The nano PPy was synthesized by microemulsion polymerization, blended with chitosan and then formed membranes. The membranes were then modified by acetylation to get the experimental membranes（nano PPy/chitin composite membranes, group A）. The chitosan membranes（group B） and chitin ones（group C） modified by acetylation acted as control. Scanning electron microscopy and FT-IR spectra were used to identify the nano PPy and the membranes of each group. And the conductivity of membranes of each group was measured. Schwann cells were cocultured in vitro with each group membranes to observe the biocompatibility by inverted microscope observing, living cell staining, cell counting, and immunofluorescence staining. The lysozyme solution was used to evaluate the degradation of the membranes in vitro. Results The FT-IR spectra showed that the characteristic vibrational absorption peaks of C=C from nano PPy appeared at 1 543.4 cm^（–1） and 1 458.4 cm^（–1）. Scanning electron microscopy observation revealed that the size of nano PPy particles was about 100-200 nm. The nano PPy particles were synthesized. It was successful to turn chitosan to chitin by the acetylation, which was investigated by FT-IR analysis of membranes in groups A and C. The characteristic peaks of the amide Ⅱ band around 1 562 cm^（–1） appeared after acetylated modification. Conductivity test showed that the conductivity of membranes in group A was about（1.259 2±0.005 7）×10–3 S/cm, while the conductivity of the membranes in groups B and C was not detected. The nano PPy particles uniformly distributed on the surface of membranes in group A were observed by scanning electron microscope; the membranes in control groups were smooth. As a result, the nano PPy/chitin composite membranes with electrical conductivity were obtained. The cultured Schwann cells were found to survive with good function by fluorescei

关 键 词：纳米 聚吡咯 壳聚糖 甲壳素 乙酰化 生物相容性 

分 类 号：R318.08[医药卫生—生物医学工程]