改性纳米生物玻璃水凝胶的制备及特性初步研究  

Preparation and preliminary research on the characteristics of modified nano-bioglass hydrogel

作　　者：韦丽春 张熠杰 黄沙[2] 姚斌[3] 李想[4] 陈旭源 黎彦 付小兵[3] 吴旭[1] Wei Lichun;Zhang Yijie;Huang Sha;Yao Bin;Li Xiang;Chen Xuyuan;Li Yan;Fu Xiaobing;Wu Xu(Department of Thoracic Surgery,Huiqiao Medical Center,Nanfang Hospital of Southern Medical University,Guangzhou 510515,China;Institute of Basic Medical Sciences,General Hospital of PLA,Beijing 100039,China;Key Laboratory of Tissue Repair and Regeneration of PLA,Beijing Key Research Laboratory of Skin Injury,Repair and Regeneration,Fourth Medical Center,General Hospital of PLA,Beijing 100048,China;Department of Emergency,Nanfang Hospital of Southern Medical University,Guangzhou 510515,China;Department of Comprehensive Medical Treatment Ward,Nanfang Hospital of Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学南方医院惠侨医疗中心胸外科,广州510515 [2]解放军总医院基础医学研究所,北京100039 [3]解放军总医院第四医学中心全军创伤修复与组织再生重点实验室暨皮肤损伤修复与再生北京市重点实验室,100048 [4]南方医科大学南方医院急诊科,广州510515 [5]南方医科大学南方医院综合科,广州510515

出　　处：《中华烧伤杂志》2020年第10期930-938,共9页Chinese Journal of Burns

基　　金：国家自然科学基金(81571909、81701906、81830064、81721092、81671905);国家重点研究发展规划(2017YFC1103300);军队后勤科研重大项目(AWS17J005);解放军总医院杰出青年培育专项(2017-JQPY-002)。

摘　　要：目的初步探讨改性纳米生物玻璃水凝胶的制备以及它的理化、生物学特性。方法(1)取400 mL氢氧化钙饱和溶液,加入纳米二氧化硅悬液67 mL,制备纳米生物玻璃悬液,观察其悬浮稳定性。(2)制备终质量分数为10%明胶、1%海藻酸钠的水凝胶,设为对照组;在对照组水凝胶基础上加入实验(1)制备的纳米生物玻璃悬液,制备终质量分数为0.5%生物玻璃、10%明胶、1%海藻酸钠的水凝胶,设为实验组。观察2组水凝胶在4、25℃的成胶情况并记录成胶时间及在37℃的熔解情况并记录熔胶时间。另取2组水凝胶,4℃冷浴后用25 g/L的氯化钙溶液交联,用杨氏模量测定仪测量压缩模量。另取2组水凝胶,同前交联后于-20℃冻干,测量相关体积并计算孔隙率。样本数均为3。(3)取12只24 h龄C57BL/6J小鼠乳鼠,分离培养成纤维细胞(Fb),倒置显微镜下观察其形态,并培养第3代Fb,用于后续实验。取Fb,制备细胞浓度为1×10^5个/mL的单细胞悬液,按随机数字表法(下同)分为实验组和对照组,分别加入实验(2)制备的实验组和对照组液态水凝胶,培养12、24、48 h,每组各取3孔,采用细胞计数试剂盒8法检测细胞存活率。(4)取第3代Fb,制备细胞浓度为(3.0~4.5)×10^7个/mL的单细胞悬液,分为实验组和对照组,每组1管,加入绿色荧光探针DIO染色,分别加入实验(2)中制备的实验组和对照组液态水凝胶9 mL,同前交联后制备载细胞水凝胶。培养3 d,激光共聚焦显微镜下观察细胞在凝胶中的存活情况;同前制备载细胞水凝胶块但不加绿色荧光探针DIO,培养7 d,在扫描电子显微镜下观察细胞在水凝胶中的黏附及伸展情况。(5)取6周龄雌性BALB/c-nu裸鼠12只,分为实验组和对照组,每组6只,在背部制作直径为1 cm的圆形全层皮肤缺损创面,伤后即刻,分别放入1块实验(4)制备的实验组和对照组载细胞水凝胶块。伤后7、14 d,每组取3只裸鼠,收集创面及创周组�Objective To explore the preparation and preliminary research on the characteristics of modified nano-bioglass hydrogel.Methods(1)The nano-bioglass suspension was prepared by adding 67 mL nano-silica suspension into 400 mL saturated calcium hydroxide solution,and its suspension stability was observed.(2)The hydrogel with final mass fraction of 10%gelatin and 1%sodium alginate was prepared and set as control group.On the basis of the hydrogel in control group,the nano-bioglass suspension prepared in experiment(1)was added to prepare the hydrogel with the final mass fraction of 0.5%bioglass,10%gelatin,and 1%sodium alginate,and the hydrogel was set as the experimental group.The gelling time at 4 and 25℃and the dissolution time at 37℃of hydrogel in 2 groups were recorded,and the gelation at 4 and 25℃and dissolution condition at 37℃of the hydrogel in 2 groups were observed.The hydrogel in 2 groups were collected and cross-linked with 25 g/L calcium chloride solution after cold bath at 4℃,and the compression modulus was measured by Young′s modulus tester.In addition,the hydrogel in 2 groups were collected and cross-linked as before,and freeze-drying hydrogel was made at -20℃.The relative volumes were measured and the porosity of hydrogel in 2 groups was calculated.The number of sample in the experiment was 3.(3)Fibroblasts(Fbs)were isolated and cultured from 12 C57BL/6J mice of 24 hours old and the morphology was observed by inverted microscope,and the third passage of Fbs were cultured for the following experiment.Fbs were collected to make single cell suspension with the cell concentration of 1×10^5/mL.The single cell suspension was divided into experimental group and control group according the random number table(the same grouping method below),which were added with hydrogel in experimental group and control group prepared in experiment(2),respectively.At culture hour 12,24,and 48,cells of 3 wells in each group were collected to detect the survival rate by cell counting kit 8 method.(4)The third pass

关 键 词：伤口愈合 生物相容性材料 水凝胶 纳米生物玻璃 

分 类 号：R644[医药卫生—外科学] Q493[医药卫生—临床医学] Q441[生物学—生理学]