静电纺丝技术制备壳聚糖-胶原纳米纤维膜及其引导颅骨骨再生的动物实验  

Development of a novel absorbable nanofiber chitosan-collagen membrane by electrospinning and the preliminary study on guided bone regeneration

作　　者：高波[1] 李晓静[1] 林敏[1] 李杨杨 董研[1] 

机构地区：[1]浙江大学医学院附属第二医院口腔修复科,杭州310009 [2]浙江大学材料科学与工程学院,杭州310027

出　　处：《中华口腔医学杂志》2018年第2期85-91,共7页Chinese Journal of Stomatology

基　　金：浙江省自然科学基金（Y2110260）

摘　　要：目的采用静电纺丝技术制备壳聚糖-胶原纳米纤维膜（nanofiber chitosan-collagen membrane，NCM），评价其引导颅骨骨再生的效果，为纳米屏障膜的临床应用提供依据。方法用胶原、壳聚糖、聚环氧乙烷三者混合液进行静电纺丝可纺性研究，制备NCM，戊二醛蒸气交联。用扫描电镜及傅里叶变换红外光谱仪检测交联前后NCM结构及成分；甲基噻唑基四唑（methyl thiazolyl tetrazolium，MTT）实验评价小鼠胚胎成骨细胞前体细胞（MC3T3-E1）在NCM上的增殖情况。将胶原膜组（Bio-gide membrane，BGM）、加压冻干膜（compressed chitosan-collagen membrane，CCM）组和NCM-CCM组材料分别覆盖SD大鼠颅骨骨缺损（每组3只），空白对照组（3只）不覆盖屏障膜；术后6周取材分析各组新骨形成及屏障膜降解情况。结果胶原、壳聚糖和聚环氧乙烷质量分数分别为4.0%、1.0%和3.5%时，混合液可通过静电纺丝技术制备NCM，交联后结构稳定性较好，傅里叶变换红外光谱显示交联过程并未改变纤维成分。NCM有良好的细胞相容性，MC3T3-E1细胞可在NCM表面快速附着并增殖。术后6周NCM-CCM组与BGM组均成骨明显，新生骨面积百分比分别为（43.10±1.49）%、（41.36±2.60）%，两组差异无统计学意义（P〉0.05）；但均显著大于CCM组和空白对照组[分别为（33.10±1.41）%和（7.22±2.46）%]（P〈0.05）。结论合理配比胶原、壳聚糖、聚环氧乙烷后可通过静电纺丝技术制备NCM，其有较强的引导骨再生的能力。Objective To evaluate the application effect of nanofiber chitosan-collagen membrane （NCM） on guided bone regeneration （GBR）.Methods The mixture of collagen, chitosan, polyethylene oxide was used to make up the NCM by electrospinning, then the NCM was crosslinked by glutaraldehyde vapor. The physical property of the NCM was measured by scanning electron microscopy （SEM） and Fourier transform infrared spectroscopy （FTIR）. MC3T3-E1 osteoblasts were cultured on NCM to characterize the biocompatibility. The effectiveness of four groups [contrast group, Bio-gide membrane （BGM）, compressed chitosan-collagen menbrane （CCM）, NCM/CCM] on bone regeneration were evaluated in critical-sized defects （diameter=5 mm） in SD rats.Results When the mixed solution consists of 4.0% collagen, 1.0% chitosan and 3.5% polyethylene oxide, the NCM could be validly fabricated by electrospinning. After cross-linking by glutaraldehyde vapor, the tensile strength and the stability of NCM in damp was enhanced. No cytotoxicity of the NCM was detected on MC3T3-E1 osteoblasts. In vivo study showed that the new bone regeneration ratio of NCM/CCM group was [（43.10±1.49）%], and this was similar to that of the group of BGM [（41.36±2.60）%] （P〉0.05）, but higher than that of the CCM group [（33.10±1.41）%] and the contrast group [（7.22±2.46）%] （P〈0.05）.Conclusions The NCM can promote new bone regeneration effectively in GBR procedure.

关 键 词：壳聚糖 胶原 骨再生 静电纺丝 

分 类 号：R318.08[医药卫生—生物医学工程]