仿生壳聚糖基水凝胶的可控构筑及其对多能干细胞生物学行为影响的分析  

Controllable Construction of Biomimetic Chitosan-based Hydrogels and Their Effects on Biological Behavior of Pluripotent Stem Cells

作　　者：刘莉[1] 徐斌 宋嘉卓 张志民[1] 洪丽华[1] LIU Li;XU Bin;SONG Jia-Zhuo;ZHANG Zhi-Min;HONG Li-Hua(School and Hospital of Stomatology,Jilin University,Changchun 130021,China;College of Chemistry,Jilin University,Changchun 130012,China)

机构地区：[1]吉林大学口腔医学院,长春130021 [2]吉林大学化学学院,长春130012

出　　处：《分析化学》2022年第10期1520-1532,共13页Chinese Journal of Analytical Chemistry

基　　金：吉林省科技发展计划项目(No.20200404184YY)资助。

摘　　要：以壳聚糖(CS)为基质,在酸性条件下,通过对天然蛋白交联剂京尼平(GNP)上的酯基进行亲核取代反应,制备了理化性能可控、可用于诱导牙周组织再生的仿生CS水凝胶(CS-GNP)。对材料的结构和形貌进行了表征,对材料的力学性能、孔隙率、溶胀率和降解率进行了检测。通过小鼠成纤维细胞(L-929)体外活力的检测对材料的细胞毒性进行了评价。通过对牙周膜间充质干细胞(hPDLSCs)增值率的观察以及利用qPCR法对牙周膜相关基因PLAP-1、 COL1、 SCX和POSTN的mRNA表达水平的检测,分析了材料对多能干细胞生物学行为的影响。研究发现,改变GNP的配比可得到具有不同理化特征的GS-GNP。材料的机械信号影响了多能干细胞的生物学行为,改变材料的弹性模量可调控干细胞的延伸面积和增殖率。当材料弹性模量为(46.02±15.70) kPa时,与正常牙周膜细胞外基质接近,可诱导hPDLSCs向牙周膜方向分化。研究结果表明,干细胞微环境的物理信号可调控细胞的命运和分化,模拟天然的细胞外基质硬度可极大限度地调控多能干细胞的定向分化,为分析和表征细胞微环境对干细胞行为的影响提供可行的模型材料和体系。Biomimetic chitosan hydrogels(CS-GNPs) with controllable physical and chemical properties for inducing periodontal tissue regeneration were prepared by performing a nucleophilic substitution reaction on the ester group of the natural protein crosslinker genipine(GNP) with polyglucine(1-4)-2-amino-β-D glucose(chitosan, CS) as the matrix under acidic conditions. The structure and morphology of the materials were characterized, and the mechanical properties, porosity, swelling rate, and degradation rate of the material were detected. The cytotoxicities of the materials were evaluated by the viability detection of mice fibroblasts(L-929) in vitro. The effects of the materials on the biological behavior of pluripotent stem cells were analyzed by observing the proliferation rate of periodontal ligament stem cells(hPDLSCs) and detecting mRNA expression levels of periodontal ligament-related genes PLAP-1, COL1, SCX and POSTN by qPCR. The experimental results showed that CS-GNP with different physicochemical characteristics could be obtained by changing the ratio of GNP. The mechanical signals of the materials affected the biological behaviors of pluripotent stem cells and changing the elastic modulus of the materials could regulate the extension area and proliferation rates of stem cells. When the material′s modulus of elasticity was(46.02±15.7) kPa, which was close to the normal periodontal ligament extracellular matrix, the differentiation of hPDLSCs could be induced to the direction of the periodontal ligament. The results verified that the physical signals of the stem cell microenvironment could regulate the fate and differentiation of cells, and the simulation of natural extracellular matrix hardness could greatly regulate the directional differentiation of pluripotent stem cells, providing feasible model materials and systems for analyzing and characterizing the influence of the cell microenvironment on stem cell behavior.

关 键 词：仿生水凝胶 可控构筑 多能干细胞 基质刚度 生物学行为 

分 类 号：TQ427.26[化学工程] R318.08[医药卫生—生物医学工程]