改性木葡聚糖温敏水凝胶的制备及生物相容性评价  

Preparation and biocompatibility evaluation of modified thermosensitive xyloglucan hydrogels

作　　者：闫志文 李硕峰 李傲 胡梓骐 马丽桃 张二帅 李景武[3] 姚芳莲[2] 车鹏程 孙红 Yan Zhiwen;Li Shuofeng;Li Ao;Hu Ziqi;Ma Litao;Zhang Ershuai;Li Jingwu;Yao Fanglian;Che Pengcheng;Sun Hong(Key Laboratory for Chronic Diseases of Hebei Province,School of Basic Medical Sciences,North China University of Science and Technology,Tangshan 063210,Hebei Province,China;Department of Polymer Science and Engineering,School of Chemical Engineering and Technology,Tianjin University,Tianjin 300350,China;Department of Gastrointestinal Surgery,Tangshan People’s Hospital,Tangshan 063001,Hebei Province,China)

机构地区：[1]华北理工大学基础医学院,河北省慢性病重点实验室,河北省唐山市063210 [2]天津大学化工学院高分子科学与工程系,天津市300350 [3]唐山市人民医院胃肠外科,河北省唐山市063001

出　　处：《中国组织工程研究》2019年第30期4841-4847,共7页Chinese Journal of Tissue Engineering Research

基　　金：河北省自然科学基金(H2017209005),项目负责人:孙红;唐山市科技创新团队资助项目(18130211B),项目负责人:孙红;国家级大学生创新创业训练计划项目(201810081005),项目参与者:李硕峰、李傲;华北理工大学研究生创新项目(2018S26),项目负责人:闫志文;华北理工大学大学生创新创业训练计划项目(X2018009),项目参与者:李硕峰、李傲~~

摘　　要：背景:可注射温敏水凝胶能随环境温度变化发生可逆性相转变并可结合微创疗法,在术后防粘连领域得到了广泛关注,但现有的可注射温敏水凝胶在预防术后粘连方面存在生物安全性差及防粘连效果不佳等问题。目的:制备改性木葡聚糖可注射温敏水凝胶,评价其生物相容性。方法:通过温和的酶解反应脱除木葡聚糖侧链的部分半乳糖,得到具有可逆“溶胶-凝胶”转变的改性木葡聚糖可注射温敏水凝胶。通过活/死细胞染色定性检测与MTT 法定量检测改性木葡聚糖水凝胶浸提液对人肝癌细胞HepG2 和人肺腺癌细胞A549 的毒性。将人肺腺癌细胞A549 包埋于改性木葡聚糖水凝胶内,培养24 h后观察细胞形态。在抗凝兔血中分别加入改性木葡聚糖水凝胶浸提液与0.1,1 g/L 改性木葡聚糖溶液,1 h后检测溶血率。将改性木葡聚糖溶液注射至C57 小鼠(中国人民解放军军事医学科学院北京实验动物中心提供)背部皮下,跟踪材料的体内降解行为,评价植入部位及周围组织的炎性反应。动物实验获得华北理工大学实验动物伦理委员会批准,批准号:SYXK(冀)2015-0038。结果与结论:①AO/PI 染色与MTT 检测显示,不同浓度改性木葡聚糖水凝胶浸提液中的HepG2 细胞、A549细胞增殖活性均在100%左右;②包埋在改性木葡聚糖水凝胶内的A549 细胞仍有很好的活性;③改性木葡聚糖水凝胶浸提液与0.1,1 g/L 改性木葡聚糖溶液的溶血率均在2%以下;④随着注射时间的延长,动物体内的改性木葡聚糖水凝胶逐渐变小,约在6 周时降解完全;注射早期可引起炎症反应,但随着时间延长与材料的降解,炎症反应逐渐减轻和消失,注射部位未见明显组织病变损伤;⑤结果表明,改性木葡聚糖水凝胶具有温敏特性,还有良好的细胞相容性、血液相容性与生物降解性。BACKGROUND: Injectable thermosensitive hydrogels have reversible phase transition with the change of ambient temperature and can be used in combination with minimally invasive therapy. This has attracted extensive attention in the field of postoperative adhesion prevention. However, the current injectable thermosensitive hydrogels for the prevention of postoperative adhesion have the problems of poor biosafety and anti-adhesion effects. OBJECTIVE: To prepare a kind of hydrogel with thermosensitive and injectable characters with the modified xyloglucan and to evaluate its biocompatibility. METHODS: Partial galactose of the side chain of xyloglucan was removed using mild enzymatic hydrolysis to obtain modified xyloglucan with reversible “sol-gel” transformation. The toxicity of modified xyloglucan hydrogel leaching liquor to human hepatoma cell line HepG2 and human lung adenocarcinoma cell line A549 was determined by acridine orange and propidium iodide staining and MTT assay. After 24 hours of culture, cell morphology was observed. Modified xyloglucan hydrogel leaching liquor, 0.1 and 1 g/L modified xyloglucan solution was added to the anticoagulated rabbit blood. The hemolysis rate was detected 1 hour later. The modified xyloglucan solution was subcutaneously injected into the back of the C57 mice (provided by Beijing Experimental Animal Center of the Chinese Academy of Military Medical Sciences, China) to track the in vivo degradation behavior of the material and evaluate the inflammatory response of the implant site and surrounding tissues. Animal experiments were approved by the Animal Ethics Committee, North China University of Science and Technology, China (approval No. SYXK (Ji) 2015-0038). RESULTS AND CONCLUSION: The acridine orange and propidium iodide staining and MTT assay revealed that the proliferation activity of HepG2 cells and A549 cells in different concentrations of modified xyloglucan hydrogel leaching liquor was about 100%. A549 cells embedded in modified xyloglucan hydrogel still had good

关 键 词：改性木葡聚糖 木葡聚糖 可注射温敏水凝胶 生物相容性 术后粘连 血液相相容性 生物材料 人肝癌细胞 人肺腺癌细胞 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]