新型壳聚糖-胶原支架与牙周膜细胞的生物相容性  

Biocompatibility of a novel chitosan-collagen scaffold and periodontal ligament cells

作　　者：唐倩[1] 孙文娟[1] 黄南楠[1] 陈茜[1] 任力 侯思润 

机构地区：[1]中山大学附属第三医院口腔科,广东省广州市510630 [2]国家人体组织功能重建工程技术研究中心,广东省广州市510641

出　　处：《中国组织工程研究》2013年第38期6766-6772,共7页Chinese Journal of Tissue Engineering Research

基　　金：广东省科技计划项目(2011B031300011;2010B031100021)~~

摘　　要：背景:目前胶原作为牙周组织工程支架材料仍具有机械强度差、降解速度快等缺点,将其与壳聚糖复合可改善上述问题。目的:评估新型壳聚糖-胶原支架材料的体外生物相容性。方法:通过MTT法评估100%,75%,50%,25%壳聚糖-胶原支架材料浸提液对人牙周膜细胞的毒性。选择第4-6代生长状态良好的人牙周膜细胞与壳聚糖-胶原支架共培养,观察细胞在支架上的生长情况,并检测与壳聚糖-胶原支架复合培养前后人牙周膜细胞碱性磷酸酶活性的变化。结果与结论:新型壳聚糖-胶原支架具有双层结构,一侧表面致密,一侧表面疏松多孔。MTT法检测不同浓度材料浸提液毒性评级为0或1级。扫描电子显微镜及组织学观察可见细胞在壳聚糖-胶原支架上增殖良好,且致密层可起屏障膜作用,阻挡细胞进入支架内部;复合培养24 h后,人牙周膜细胞的碱性磷酸酶活性与复合培养前无明显差异(P>0.05),复合培养48,72 h后人牙周膜细胞的碱性磷酸酶活性高于复合培养前(P<0.05)。以上结果提示新型壳聚糖-胶原支架具有良好的生物相容性及屏障功能,可进一步应用于牙周组织工程的研究。BACKGROUND：At present, col agen as a material in periodontal tissue engineering has some disadvantages such as poor mechanical strength and rapid degradation speed. Col agen combined with chitosan can improve above-mentioned problems. OBJECTIVE：To evaluate the biocompatibility of a novel chitosan-col agen scaffold in vitro. METHODS：Cytotoxicity of the extract of chitosan-col agen scaffold in different concentrations （100%, 75%, 50%, 25%） was evaluated by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. Human periodontal ligament cel s at 4-6 passages were cocultured with the chitosan-col agen scaffold. Cel growth on the scaffold was observed. Changes in alkaline phosphatase activity were detected in periodontal ligament cel s before and after coculture. RESULTS AND CONCLUSION：The novel chitosan-col agen scaffold was made up of double layers with one dense layer and another loose layer. The grade of the cytotoxicity of the scaffold was from 0 to 1. Scanning electron microscope and histological observation demonstrated that cel s grew wel on the chitosan-col agen scaffold;the dense layer could prevent cel s to migrate into the scaffold. There were no significant differences in alkaline phosphatase activity in human periodontal ligament cel s before and 24 hours after combined culture （P〉0.05）. Alkaline phosphatase activity in human periodontal ligament cel s was greatly higher at 48 and 72 hours after combined culture compared with that before culture （P〈0.05）. Above results indicated that the novel chitosan-col agen scaffold has a good biocompatibility and barrier function, and potential as a scaffold for periodontal tissue engineering.

关 键 词：生物相容性材料 口腔 壳聚糖 胶原 牙周膜 

分 类 号：R318[医药卫生—生物医学工程]