生物工程学报ChinJBiotech2008,August25;24(8):1381-1386journals.im.ac.cnChineseJournalofBiotechnologyISSN1000-3061cjb@im.ac.cn©2008InstituteofMicrobiology,CAS&CSM,AllrightsreservedReceived:November26,2007;Accepted:January3,2008Supportedby:theNationalHighTechnologyResearchandDevelopmentProgramofChina(Nos.2006AA02A132,2007AA091603).Correspondingauthor:WanshunLiu.Tel:+86-532-82032105;E-mail:WanshunLiu@hotmail.com国家“十一五”863项目(Nos.2006AA02A132,2007AA091603)资助。研究报告壳聚糖基角膜细胞载体的制备及其细胞相容性高兴爽,刘万顺,韩宝芹,位晓娟中国海洋大学海洋生命学院,青岛266003摘要:为探讨羟丙基壳聚糖基共混膜作为组织工程技术中角膜细胞培养载体的可行性,分别制备了羟丙基壳聚糖/硫酸软骨素、羟丙基壳聚糖/明胶/硫酸软骨素以及羟丙基壳聚糖/氧化透明质酸/硫酸软骨素三种共混膜。测定其透光率、含水量和蛋白吸附性能;在共混膜上培养兔角膜上皮细胞,通过观察角膜上皮细胞在不同载体膜上的生长状态、贴附情况,测定细胞活性以及上清液中乳酸脱氢酶的活性,研究三种壳聚糖基载体膜片与角膜上皮细胞的相容性。膜片理化性质测定结果表明三种共混膜片具有良好的透明度,适宜的含水量和较强的蛋白吸附性能;细胞相容性实验结果表明羟丙基壳聚糖/明胶/硫酸软骨素共混膜对细胞的损伤最小,有利于细胞在膜上的贴附和生长,表现出良好的细胞相容性,有望作为角膜细胞载体体外构建组织工程化角膜。关键词:壳聚糖,载体,角膜上皮细胞,细胞相容性PreparationandCytocompatibilityofChitosan-basedCarriersofCornealCellsXingshuangGao,WanshunLiu,BaoqinHan,andXiaojuanWeiCollegeofMarineLifeSciences,OceanUniversityofChina,Qingdao266003,ChinaAbstract:Tostudythepossibilityofusinghydroxypropylchitosan-basedblendmembranesascarriersofcornealcellsintissueengineering,wepreparedthreekindsofblendmembraneslabeledhydroxypropylchitosan/chondroitinsulfate,hydroxypropylchitosan/gelatin/chondroitinsulfateandhydroxypropylchitosan/oxidizedhyaluronicacid/chondroitinsulfate.Thetransparency,watercontentandabilityofproteinadsorptionoftheblendmembranesweremeasured.Toevaluatethecytocompatibilityoftheblendmembraneswithcornealepithelialcells,rabbitcornealepithelialcellswereculturedonthesurfaceofthecarriermembranes.Themorphologicalcharacteristics,celladhesion,cellproliferationandtheactivityoflactatedehydrogenase(LDH)inthemediawereinvestigated.Threekindsofblendmembraneshadgoodopticaltransmittance,suitablewatercontentandabilityofproteinadsorption.Theresultsshowedthatthelessinjurywasmadetocornealepithelialcellsbythehydroxypropylchitosan/gelatin/chondroitinsulfateblendmembranethantheothers.Thiskindofmembranewasfavorofthegrowthandadhesionofcornealepithelialcells.Thehydroxypropylchitosan/gelatin/chondroitinsulfateblendmembraneisapromisingcarrierofcornealcellsandcanbeusedinreconstructionoftissueengineeredcornea.Keywords:chitosan,carrier,cornealepithelialcell,cytocompatibility1382ISSN1000-3061CN11-1998/QChinJBiotechAugust25,2008Vol.24No.8Journals.im.ac.cn角膜是眼球光学系统的重要组成部分,因各种疾患或创伤引起的角膜病变,可导致患者视力下降甚至致盲。有资料表明,临床上角膜疾患占眼科所有疾病的40%,其中80%可以通过角膜移植术恢复透明[1]。但角膜供体来源的短缺和移植后免疫排斥反应的发生是困扰医生和患者的最大问题。随着组织工程技术的发展和角膜细胞组织培养方法的建立,构建组织工程角膜进行移植来治疗角膜盲成为研究的一个热点。迄今为止,被研究用作角膜细胞培养贴附载体的材料包括:异体角膜片、明胶薄膜、水凝胶膜、胶原基质、可降解聚合物膜、羊膜以及后弹力层膜等[2−7],但降解速度过慢或过快,机械强度不足以及免疫原性等问题,制约了其用于临床的...
