摘要
目的制备一种可用于修复伴有骨关节炎的软骨缺损的兼具软骨再生和炎性微环境调控的水凝胶支架。方法通过溶剂热法制备锆金属有机框架(Zr MOF),并负载抗炎肽KAFAK,用紫外光交联法制备普鲁兰多糖/聚乙二醇复合水凝胶,将Zr MOF封装于水凝胶内部,通过扫描电镜、溶胀平衡、降解实验和流变力学表征水凝胶的理化性能。以兔骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)作为种子细胞,用活/死细胞染色验证水凝胶的生物活性,测定软骨细胞外基质糖胺聚糖的含量和成软骨标志基因COL2A1和ACAN及肥大化基因COL10A1的表达。结果成功将抗炎肽KAFAK负载到Zr MOF,水合粒径为352.5 nm,粒径分布(PDI)为0.29,Zeta电位为(18.27±0.55)mV。普鲁兰多糖/聚乙二醇复合水凝胶封装Zr MOF后,随着Zr MOF含量的增加,扫描电镜观察到孔壁上的小孔越多;溶胀时间缩短,降解速度增加,流变性能改善。抗炎肽KAFAK负载到Zr MOF后再用水凝胶封装,到第21天,有(84.1±5.0)%的释放,显示出对抗炎肽KAFAK较好的缓释作用。活细胞在Gel、Gel-100 Zr MOF和Gel-100 Zr MOF@KAFAK组的存活比率分别为(90.7±5.4)%、(93.5±4.3)%和(91.9±3.1)%,均显示出良好的生物相容性。以兔BMSCs为种子细胞,在成软骨诱导培养7 d和14 d后,Gel-100 Zr MOF@KAFAK组相较于Gel和Gel-100 Zr MOF组糖胺聚糖含量有显著提高(P<0.05),成软骨标志基因COL2A1、ACAN表达显著升高(P<0.05),而肥大化基因COL10A1表达显著降低(P<0.05)。结论成功将负载抗炎肽的Zr MOF封装于水凝胶内部,通过对抗炎肽KAFAK的持续释放控制炎性微环境,产生更多的软骨细胞外基质,有利于维持成软骨表型。
Objective To prepare a hydrogel scaffold with cartilage regeneration and inflammatory microenvironment control capacity for chondrodefects treatment in osteoarthritis.Methods The zirconium metal-organic framework(Zr MOF)was prepared by solvothermal synthesis and then loaded with anti-inflammatory peptide KAFAK.The pullulan/polyethylene glycol composite hydrogel was prepared by UV crosslinking.Then the obtained Zr MOF was encapsulated in the hydrogel.Then the physicochemical properties of the hydrogel were characterized by scanning electron microscopy(SEM),swelling equilibrium,degradation experiments and rheological and mechanical studies.Rabbit bone mesenchymal stem cells(BMSCs)were used as seed cells.The bioactivity of the hydrogel and the content of glycosaminoglycans in the extracellular matrix were tested by live/dead staining.The expression of chondrogenic genes COL2 A1 and ACAN,and hypertrophic gene COL10 A1 was measured with RT-PCR.Results After being loaded with anti-inflammatory peptide KAFAK to Zr MOF,the hydrated particle exhibits a size of 352.5 nm,a particle size distribution(PDI)of 0.29,and a zeta potential of 18.27±0.55 mV.After the encapsulation of Zr MOF,when the content of Zr MOF was increased,there were more small holes in the scaffold observed by SEM,the swelling time was shortened,the degradation rate was increased and the rheological property was improved.In 21 d after KAFAK was loaded onto the Zr MOF and then encapsulated with hydrogel,the release of the peptide was(84.1±5.0)%,indicating a good sustained release.In Gel,Gel-100 Zr MOF,and Gel-100 Zr MOF@KAFAK groups,the cell survival rates was(90.7±5.4)%,(93.5±4.3)%and(91.9±3.1)%,respectively.With rabbit BMSCs as seed cells,after 7 and 14 d of chondrogenic induction,the glycosaminoglycan in Gel-100 Zr MOF@KAFAK group was significantly higher than that in Gel and Gel-100 Zr MOF group(P<0.05).The results of RT-PCR indicated that the expression levels of COL2 A1 and ACAN were increased,whereas that of COL10 A1 was decreased(P<0.05).Conclus
作者
秦晓平
李涛
陈诚
杨柳
QIN Xiaoping;LI Tao;CHEN Cheng;YANG Liu(Center for Joint Surgery,First Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400038;College of Medical Informatics,Chongqing Medical University,Chongqing,400016,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2022年第7期673-683,共11页
Journal of Army Medical University
关键词
水凝胶
细胞外基质
炎性微环境
抗炎肽KAFAK
金属有机框架
hydrogels
extracellular matrix
inflammatory microenvironment
anti-inflammatory peptide KAFAK
metal-organic frameworks
