摘要
目的评价壳聚糖/明胶静电纺丝膜对球结膜损伤后组织的促修复作用及抗瘢痕化作用。方法利用静电纺丝技术制备壳聚糖/明胶静电纺丝膜，采用钨灯丝扫描电子显微镜观察静电纺丝膜纤维表征。采用随机数字表法将30只新西兰白兔随机分为模型组、羊膜修复组和静电纺丝膜修复组，每组10只。各组实验兔右眼均接受手术切除6 mm×8 mm的球结膜，用弱酸灼伤损伤部位和角膜缘，制备兔球结膜损伤动物模型，羊膜修复组和静电纺丝膜修复组均于术中在伤口处结膜与巩膜之间平铺生物膜。分别于术后4、8、14、28 d行裂隙灯显微镜检查，观察兔眼手术伤口炎症反应及球结膜修复情况。结果扫描电子显微镜下可见静电纺丝膜纤维呈立体网状结构，与生物羊膜空间结构相似，纺丝直径均匀，为0.1～0.5 μm。裂隙灯显微镜下观察发现，术后4 d各组兔实验眼均未见明显炎症反应；术后8 d模型组兔眼球结膜水肿、糜烂，黏性分泌物较多，羊膜修复组和静电纺丝膜修复组兔球结膜轻度充血；术后14 d模型组兔球结膜水肿、充血，羊膜修复组和静电纺丝膜修复组兔球结膜均未见炎症反应；术后28 d模型组兔球结膜可见新生血管，羊膜修复组和静电纺丝膜修复组兔球结膜完全修复。组织病理学观察结果显示，模型组兔眼在术后4 d时伤口处球结膜组织游离，术后8 d球结膜根部贴附巩膜，术后14 d球结膜组织增厚，术后28 d大量纤维组织填充伤口，结膜上皮化。羊膜修复组兔眼术后4 d羊膜贴附于巩膜与球结膜之间，术后8 d羊膜开始消融，术后14 d纤维组织生长且球结膜上皮化，术后28 d球结膜组织修复，瘢痕组织填充较少。术后4 d静电纺丝膜修复组可见静电纺丝膜孵育于结膜与巩膜之间，静电纺丝膜消融，术后8 d球结膜上皮沿巩膜长入损伤部位，术后14 d结膜下组织生长，术后28 d结膜完全修复。结论与单纯球结膜损伤缝合相比，羊膜或壳聚糖/明胶静电纺丝膜修补对损伤的球结膜有促进修复的作用，且可减轻组织瘢痕化，壳聚糖/明胶静电纺丝膜的促组织修复作用优于羊膜。
ObjectiveTo investigate the promoting repair effects of chitosan/gelatin electrospinning membrane implantation on conjunctival injury.MethodsChitosan/gelatin electrospinning membrane was prepared using electrospinning technique, and the general characteristics of the membrane was examined under the scanning electron microscope.Conjunctival injury models were established in the right eyes of 30 New Zealand white rabbits by surgical resection of 6 mm×8 mm bulbar conjunctiva, burns of weak acid at injury site and then randomized into model group, human amniotic membrane （AM） implanted group and electrospinning membrane implanted group.AM or electrospinning membrane was interlayerly implanted between conjunctiva and sclera during operation according to grouping.The inflammatory response and histopathological change at injury site were examined by slit lamp microscope and optical microscope in 4, 8, 14 and 28 days after surgery.The use and care of the animals followed by code of ethics for animal experiments at the animal experiment center of He University.ResultsChitosan/gelatin electrospinning membrane presented with a crosslinked network structure like AM, with the nano-microstructure of 0.1-0.5 μm fibre.No obvious inflammatory symptom was seen in the experimental eyes of various groups in 4 days after operation.In 8 days after operation, conjunctival edema and erosion were found in the model group, and slight conjunctival congestion was exhibited in the eyes of AM implanted group and electrospinning membrane implanted group.In 14 to 28 days after operation, conjunctival edema and new blood vessels were seen in the model group, and the conjunctivas were well recovered in the AM implanted group and electrospinning membrane implanted group.Histopathological examination showed dissociation, attaching, thickening and epithelization of injuried conjunctivas gradually in the model group from 4 to 28 days after operation.In the AM implanted group, attaching of AM was in 4 days, melting of AM was in 8 days, epithelization of injuried conjunctivas was in 14 days and renovation of conjunctivas was in 24 days after operation, and a rapid repairing procedure was found in the electrospinning membrane implanted group under the optical microscope.ConclusionsCompared with the self-repairing process of conjunctival injury, implantion of chitosan/gelatin electrospinning membrane or AM can speed up the renovation of injuried conjunctivas, and the effects of chitosan/gelatin electrospinning membrane are better than those of AM in promoting conjunctiva repair and inhibiting scarring.
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2018年第2期102-106,共5页
Chinese Journal Of Experimental Ophthalmology
基金
辽宁省自然科学基金项目（2014020153）
沈阳市科技局计划项目（F13-081-200）
关键词
球结膜损伤
结膜重建
生物材料
壳聚糖
明胶
静电纺丝膜
羊膜
动物模型
Injury, conjunctival
Conjunctival reconstruction
Biomaterials
Chitosan
Gelatin
Electrosinning chitosan membrane
Amniotic membrane
Disease models, animal
