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作 者:赵思楠 宫宁 张致浩 徐磊[1,2] 谈飞[1] ZHAO Si-nan;GONG Ning;ZHANG Zhi-hao;XU Lei;TAN Fei(Department of Prosthodontics,the Affiliated Hospital of Qingdao University,Qingdao 266003,China;School of Stomatology,Qingdao University,Qingdao 266003,China)
机构地区:[1]青岛大学附属医院口腔修复科,青岛266003 [2]青岛大学口腔医学院,青岛266003
出 处:《口腔颌面修复学杂志》2024年第3期161-169,共9页Chinese Journal of Prosthodontics
基 金:山东省自然科学基金面上项目(项目编号:ZR2022MH239)。
摘 要:目的:制备4组含不同种类二价金属离子Ca^(2+)、Sr^(2+)、Mg^(2+)、Zn^(2+)交联的藻酸盐凝胶支架,检测其理化性能,观察MC3T3-E1细胞在各组凝胶表面的黏附、增殖及成骨分化情况。方法:配制4组离子总浓度为6.5g/L且含不同种类Ca^(2+)、Sr^(2+)、Mg^(2+)、Zn^(2+)的混合溶液作为交联剂,与海藻酸钠溶液发生螯合反应制备多离子交联水凝胶,依据所含离子种类分为Ca组、Ca-Sr组、Ca-Sr-Mg组、Ca-Sr-Mg-Zn组,Ca组为对照组。使用扫描电镜观察凝胶冻干后的表面形态,检测其降解、溶胀性能及Sr^(2+)、Mg^(2+)、Zn^(2+)析出浓度。将MC3T3-E1细胞接种在水凝胶表面,观察细胞黏附形态,采用CCK-8法和活死染色法评价其生物安全性和细胞增殖能力。诱导MC3T3-E1细胞成骨分化,检测ALP活性,茜素红染色观察钙盐沉积情况,RT-qPCR检测成骨基因的表达。结果:与Ca组相比,Ca-Sr组、Ca-Sr-Mg组、Ca-Sr-Mg-Zn组水凝胶的孔隙更均匀致密,凝胶的降解速度与溶胀率均减小(P<0.05),细胞在凝胶表面的黏附形态由球形变为多角形;与Ca组比较,Ca-Sr组、Ca-Sr-Mg组、Ca-Sr-Mg-Zn组水凝胶表面的细胞增殖活性、细胞内ALP活性、钙结节生成面积及I型胶原、骨形态发生蛋白2、Runt相关转录因子2等成骨基因的表达量升高(P<0.05),且Ca-Sr组、Ca-Sr-Mg组、Ca-Sr-Mg-Zn组水凝胶表面的细胞增殖活力及成骨分化效果依次提高(P<0.05)。结论:较Ca^(2+)交联的海藻酸钙水凝胶相比,使用Ca^(2+)、Sr^(2+)、Mg^(2+)、Zn^(2+)混合金属阳离子作为交联剂,能改善藻酸盐水凝胶的理化性能,促进MC3T3-E1细胞在凝胶表面的黏附及成骨分化。Objective:This study aimed to produce alginate gel scaffolds crosslinked with different divalent metal ions(Ca^(2+),Sr^(2+),Mg^(2+)and Zn^(2+))and evaluate their physiochemical and biological properties.Methods:The hydrogels were prepared by reacting sodium alginate solution with mixed ions(Ca^(2+),Sr^(2+),Mg^(2+)and Zn^(2+))as cross-linking agents at a total concentration of 6.5 g/L.The gels were divided into four groups:Ca,Ca-Sr,Ca-Sr-Mg and Ca-Sr-Mg-Zn,with Ca group serving as the control.The physicochemical properties,including surface morphology,degradation rate,ion precipitation concentration and dissolution rate of the hydrogels were investigated.MC3T3-E1 cells were then seeded on the hydrogels to evaluate their effects on cell proliferation and osteogenesis.Results:Compared to the Ca group,the Ca-Sr,Ca-Sr-Mg and Ca-Sr-Mg-Zn groups exhibited more uniformed and dense hydrogel pores,as well as reduced degradation and swelling rates(P<0.05).The cells adhered to the gel surface also exhibited a change in morphology from spherical to polygonal.The experimental group demonstrated increased cell proliferation,viable cell count,intracellular ALP activity,calcium nodule formation area,and expression of osteogenic genes(Col-1,BMP-2,Runx-2)compared to the Ca group(P<0.05).The proliferative and osteogenic effects on the gel surface increased sequentially in the Ca-Sr,Ca-Sr-Mg and Ca-Sr-Mg-Zn groups(P<0.05).Conclusion:The use of a mixture of Ca^(2+),Sr^(2+),Mg^(2+)and Zn^(2+)as cross-linking agents improves the physiochemical properties of alginate hydrogels and enhances cell adhesion and osteogenic differentiation of MC3T3-E1 cells on the gel surface compared to single Ca^(2+)cross-linked calcium alginate hydrogels.
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