玉米壳纤维增强壳聚糖基水凝胶应用于软骨组织工程支架的潜力  

Potential of corn husk fibers reinforced chitosan-based hydrogels in cartilage tissue engineering scaffold

作　　者：方妤露 易兵成 沈炎冰[1] 唐寒 张彦中[1] Fang Yulu;Yi Bingcheng;Shen Yanbing;Tang Han;Zhang Yanzhong(College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,Shanghai 201620,China)

机构地区：[1]东华大学化学化工与生物工程学院,上海市201620

出　　处：《中国组织工程研究》2020年第34期5493-5501,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(31771050,31570969),项目负责人:张彦中。

摘　　要：背景:如何增强水凝胶的力学性能是软骨组织工程领域的一个研究热点。课题组前期采用柠檬酸对壳聚糖水凝胶进行改性,显著提升了改性后的水凝胶在湿态下的拉伸和压缩力学性能。目的:观察用天然玉米壳来源的纤维素纤维增强柠檬酸改性壳聚糖水凝胶的力学提升效果,以及其作为软骨组织工程支架的潜力。方法:采用冷冻-解冻法分别制作柠檬酸改性壳聚糖水凝胶与纤维素纤维/柠檬酸改性壳聚糖复合水凝胶(其中纤维素纤维与柠檬酸改性壳聚糖的质量比分别为1∶1和2∶1),表征这些水凝胶的结构与力学性能。将兔软骨细胞接种于3种水凝胶表面,通过活死细胞染色、CCK-8检测、扫描电镜等方法分析水凝胶的细胞相容性,通过胶原免疫荧光染色与糖胺聚糖定量检测分析水凝胶促软骨细胞功能表达的能力。结果与结论:①随着纤维素纤维加入比例的增加,柠檬酸改性壳聚糖水凝胶的孔隙率、平衡溶胀率、降解速率逐渐降低,干态或湿态下的抗压强度、杨氏模量逐渐升高;②活死细胞染色显示,接种7 d后软骨细胞在3种水凝胶表面增殖良好;③CCK-8检测显示,随着纤维素纤维加入比例的增加,柠檬酸改性壳聚糖水凝胶中的细胞增殖逐渐降低;④扫描电镜观察显示,柠檬酸改性壳聚糖水凝胶上的软骨细胞保持椭圆形,呈单个、稀疏地分布在水凝胶片层表面;在纤维素纤维/柠檬酸改性壳聚糖复合水凝胶上的软骨细胞多数呈完全铺展的形貌;⑤接种28 d后,随着纤维素纤维加入比例的增加,柠檬酸改性壳聚糖水凝胶上软骨细胞的Ⅰ型胶原表达逐渐降低,Ⅱ型胶原表达逐渐升高,而糖胺聚糖含量无明显变化;⑥结果表明,纤维素纤维/柠檬酸改性壳聚糖复合水凝胶具有较好的机械强度,有作为软骨组织工程支架的应用潜力。BACKGROUND:How to enhance the mechanical properties of the chitosan-based hydrogels has become a focus in the field of cartilage tissue engineering.In our previous research,chitosan was modified by citric acid,which led to significant improvement in the tensile and compression strength of the modified hydrogen in wet state.OBJECTIVE:To further enhance the mechanical performance of the citric acid modified chitosan hydrogel(i.e.,CC)using the cellulose fibers derived from the natural corn husk fibers(CHFs),and to explore the potential of CHFs/CC composite hydrogel for cartilage tissue engineering scaffold.METHODS:A freeze-thawing method was employed to prepare CC and CHFs/CC(mass ratios of cellulose fiber to citric acid modified chitosan:1:1 and 2:1).The structure and mechanical properties of the prepared hydrogels were characterized.Rabbit chondrocytes were seeded onto the hydrogel surface,and the cytocompatibility of the prepared hydrogels was analyzed by live/dead cell staining,CCK-8 assay,and scanning electron microscopy.The ability of CHFs/CC composite hydrogels in promoting functional expression of chondrocytes was determined by collagen immunofluorescence staining and quantitative analysis of glycosaminoglycans.RESULTS AND CONCLUSION:(1)With the increase in CHFs loading fractions,the porosity,equilibrium swelling rate,and degradation rate of CHFs/CC composite hydrogels were gradually reduced,but the compressive strength and Young’s modulus in dry and wet states were significantly enhanced.(2)Live/dead cell staining revealed that at 7 days of culture,chondrocytes proliferated well on the surface of three types of hydrogels.(3)The CCK-8 assay showed that with the increase in CHF loading fractions,the proliferative ability of chondrocytes on CC was gradually decreased.(4)Scanning electron microscope revealed that the chondrocytes on CC remained elliptic and were sparsely distributed,whereas most of the chondrocytes on CHFs/CC showed completely spread morphology.(5)After 28 days of culture,with the increase of

关 键 词：玉米外壳 纤维素纤维 壳聚糖 水凝胶 改性 力学性能 软骨组织工程 

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