摘要
目的:研究新型组织工程纳米材料丝素蛋白/左旋聚乳酸(SF/PLLA)无纺网的生物相容性和安全性,探讨其作为生物植入材料的可行性。方法:采用静电纺丝方法制备SF/PLLA复合组织工程纳米材料,制备浸提液,并以生理盐水作为对照,注射入小鼠体内,观察2周内小鼠的一般情况和不良反应;将材料植入小鼠背部,以3-0缝线作为对照,于术后1、2、3和4周分别取材,HE染色并拍照,比较实验组与对照组周围组织的炎症反应情况;培养兔膝关节软骨细胞,传代后接种至材料表面,在体外继续培养,倒置光学显微镜观察细胞的黏附和生长情况。将关节软骨细胞分为阴性对照组(空白培养液)、实验组(含材料的培养液)和阳性对照组(含苯酚培养液),不同组材料浸提液与细胞共培养24和48h,MTT法检测材料对软骨细胞增殖的影响。结果:注射浸提液后小鼠的全身状况与注射生理盐水后无差别;材料植入小鼠背部后,开始有少量成纤维细胞长入材料中,伴随出现少量的淋巴细胞及异物巨细胞,随后成纤维细胞数量增加,淋巴细胞及异物巨细胞无明显增多,材料慢慢降解,直至第4周时材料出现明显的降解;材料周围组织炎症反应,第2周时最严重,然后逐渐减轻,其周围组织炎症反应与缝线周围组织一致或略轻;将细胞接种于支架上24h后有细胞贴附于材料纤维上,48h后贴附于纤维上的细胞数量增多。MTT法检测,实验组24和48h细胞毒性均为Ⅰ级。除阳性对照组外,其他各组随着培养时间的延长,吸光度(A)值均增加;同一时间点,实验组与阴性对照组A值比较差异无统计学意义(P>0.05),与阳性对照组比较A值明显升高(P<0.01)。结论:SF/PLLA无纺网支架材料属于安全植入性材料,具有良好的生物相容性。
Objective To study the biocompatibility of silk fibroin/poly L-lactic acid (SF/PLLA) non-woven network,a kind of new composite tissue engineering nanomaterials,and to explore its possibility as the biological implant materials.Methods The PLLA non-woven network was prepared by electrostatic spinning.Physiological saline as control,the leaching solution was prepared and injected into the mice,then the mice were observed for 2 weeks.The materials were implanted into the back of the mice,and 3-0 suture was used as control.Tissues were collected at 1,2,3,and 4 weeks after operation,dyed by HE staining and then the photos were taken.The&amp;nbsp;tissue reactions in experimental group and control group were observed.The rabbit knee joint cartilage cells were cultured,and then subculture cells were seeded to the surface of materials.After cultured invitro,the adhesion and growth of the cells were observed with inverted optical microscope.The bioactivities of the rabbit knee joint cartilage cells in negative control group(DMEM culture media),experimental group(DMEM containing materials) and positive control group(DMEM containing phenol solution)were determined by MTT assay after cocultured for 24 and 48 h.Results After injection,the body status of the mice in experimental group was the same to the control group.There were little fibroblasts was and a little of lymphocytes and macrophage cells in the materials which were implanted into the back of the mice at the beginning.Then the number of the fibroblasts was increased, but the number of the lymphocytes and macrophage cells did not change obviously.The materials degraded slowly, and the material degraded obviously at 4 weeks.The inflammation of tissue around the material reduced gradually from the 2nd week.The inflammation of tissue around the material was the same to the suture,and sometimes was slighter than the suture.After sed for 24 h,there were cells attaching to the fibers of the material.More and more cells attached to the fibers.The reasult of MTT assay showed that the cytotoxicities in experimental groups were all on LevelⅠ at 24 and 48 h.Except for positive control group,the A values were increased in other groups with the extended response time.At the same time,there was no significant difference in cytotoxicity between experimental group and negative control group(P〉0.05)and the A value in experimental group was higher than that in positive control group(P〈0.01).Conclusion The SF/PLLA non-woven network scaffold material has good biological compatibility and safety,it could be used as implant material in tissue engineering.
出处
《吉林大学学报(医学版)》
CAS
CSCD
北大核心
2014年第3期578-582,I0004,共6页
Journal of Jilin University:Medicine Edition
基金
吉林省科技厅科研基金资助课题(3D512K513431)
关键词
丝素蛋白
左旋聚乳酸
组织工程
软骨
膝关节
生物相容性
体外细胞毒性
急性全身毒性
短期局部反应
silk fibroin
poly L-lactic acid
tissue engineering
cartilage
knee joint
biocompatibility
in vitro cytotoxicity
acute general toxicity
short-term local reactions