鱼鳞胶原蛋白止血及体内降解和生物学的相关评价  

Hemostatic efficacy,degradation in vivo and biological evaluation of fish scale collagen

作　　者：陈玥 罗峰 苏丹 杨棣远 蒲琦 张敏 贵文娟 孙效容 Chen Yue;Luo Feng;Su Dan;Yang Diyuan;Pu Qi;Zhang Min;Gui Wenjuan;Sun Xiaorong(Chengdu Bio-HT Company Limited,Chengdu 610000,Sichuan Province,China;Sichuan Bio-HT Company Limited,Chengdu 610000,Sichuan Province,China;General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]成都合拓创展生物科技有限公司,四川省成都市610000 [2]四川合拓创展生物科技有限公司,四川省成都市610000 [3]宁夏医科大学总医院,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2022年第27期4353-4359,共7页Chinese Journal of Tissue Engineering Research

基　　金：四川省科技计划(2021JDRC0061),项目负责人:陈玥。

摘　　要：背景:可吸收止血材料是目前止血材料研究的热门领域,临床上对安全有效的可吸收止血材料的需求不断扩大,鱼鳞胶原蛋白作为可吸收止血材料不仅安全有效,还可将鱼鳞变废为宝,合理充分利用资源。目的:验证鱼鳞胶原蛋白的止血功效、体内降解程序及时间,并探讨其安全性。方法:通过化学脱钙联合低温酶法提取鱼鳞胶原蛋白材料,用于以下3部分实验。①第1部分为止血实验:包括体内-肝脏止血和体表-尾止血,通过物理损伤至肝脏/尾出血制备肝出血和尾出血模型,将成模动物分为实验组(使用鱼鳞胶原蛋白)、对照组(使用止血海绵)和空白组(不进行处理),记录出血量和止血时间。②第2部分为鱼鳞胶原蛋白体内降解实验:使用肝脏止血实验动物,实验组动物填充鱼鳞胶原蛋白材料后,分别于干预后1,3,5,10,20,30,40,50,60 d实施安乐死;对照组动物填充止血海绵后于干预后5,50 d实施安乐死;空白组动物造模后于5,50 d安乐死;解剖观察体内粘连情况,病理观察填充物降解情况和肝脏恢复再生进度。③第3部分为生物学评价,包括溶血实验、表皮刺激实验、皮内刺激实验、热原实验和急性毒性实验。结果与结论:①使用化学脱钙联合低温酶法,成功提取绒丝状鱼鳞胶原蛋白,得率66%-73%,鱼鳞胶原蛋白孔径为(0.80±0.23)mm;②通过尾出血模型验证体表止血功效,鱼鳞胶原蛋白与空白组相比,显著缩短止血时间、且出血量较对照组(止血海绵)和空白组显著减少(P<0.05);③鱼鳞胶原蛋白植入动物体内后5 d与受损肝脏贴合紧密,未见腹腔粘连,干预后20 d可见鱼鳞胶原蛋白体积明显缩小,干预后50 d则解剖眼观未见材料;④经病理检测可见,植入胶原蛋白后1-3 d处于急性炎症反应期;干预后5 d可见细胞附着于材料上;干预后10 d材料与肝脏交界处形成血管结构;干预后20-40 d材料中形成肝血窦样结构;干预BACKGROUND:Absorbable hemostatic mate rials are currently a hot field of hemostatic materials.The clinical demand for safe and effective absorbable hemostatic materials is increasing.Fish scale collagen is not only safe and effective,but also can turn fish scales into treasures and make full use of resources reasonably.OBJECTIVE:To verify the hemostatic efficacy,the degradation procedure in vivo,and explo re the safety of fish scale collagen.METHODS:Fish scale collagen mate rial was extra cted by chemical decalcification combined with low temperature enzymatic method,and used in the following three tests.(1) The First Part:The bleeding test included the body-liver bleeding and surfa ce-tail bleeding.Liver bleeding and tail hemorrhage model was prepared by physical damage to the liver and tail.Model animals were divided into expe rimental group(use of fish scale collagen),the control group(use of hemostatic sponge),and blank group(no processing).The amount of blood loss and hemostatic time were recorded.(2) Part Ⅱ:In vivo degradation test of fish scale collagen:Animals with liver hemostasis were euthanized at 1,3,5,10;20,30,40,50,and 60 days after being filled with fish scale collagen material.Animals in the control group were euthanized at 5 and 50 days after being filled with hemostatic sponge.Animals in the blank group we re euthanized at 5 and 50 days after modeling.The adhesions in abdominal cavity we re observed,and the degradation of material and the progress of liver regeneration were observed pathologically.(3) Part Ⅲ:Biological evaluation,including hemolysis test,epidermal stimulation test,intradermal stimulation test,pyrogen test,and acute toxicity test.RESULTS AND CONCLUSION:(1) The collagen of filamentous fish scale was extra cted successfully by chemical decalcification combined with low temperature enzymatic method;the yield was 66%-73%,and the pore size of collagen of fish scale was 0.80±0.23 mm.(2) Tail bleeding model was used to ve rify the hemostatic effect of body surfa ce.Compared with th

关 键 词：鱼鳞胶原蛋白 生物止血材料 肝出血模型 尾出血模型 止血时间 出血量 体内降解 肝脏修复 生物相容性 可吸收材料 

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