摘要
背景:课题组前期研究证实,镁基金属与矿化胶原联合应用在修复极限缺损时具有良好的支撑效果,可在一定程度上改善矿化胶原机械强度过差及在骨愈合期间过早塌陷的问题。然而,镁基金属在含氯化物溶液(包括人体体液或血浆)中降解较快,其中释放镁离子对成骨细胞增殖分化等的影响尚不清楚。目的:分析镁离子联合矿化胶原对小鼠前成骨细胞体外成骨分化能力的影响。方法:分别采用镁离子浓度为0,5,10,20 mmol/L的完全培养基制备矿化胶原浸提液,以4种矿化胶原浸提液培养小鼠前成骨细胞,分别设为矿化胶原组及5,10,20 mmol/L Mg2++矿化胶原组,以完全培养基培养的小鼠前成骨细胞为空白对照组。观察细胞的形态、增殖、凋亡、细胞内微丝肌动蛋白与成骨诱导分化后的碱性磷酸酶活性及成骨基因Runx2的表达。结果与结论:①培养24 h,矿化胶原组、5,10 mmol/L Mg2++矿化胶原组细胞贴壁情况良好,与空白对照组无明显差异,细胞形态呈长梭形,表面有多个突触与邻近的细胞相联系;20 mmol/L Mg2++矿化胶原组细胞呈现明显的核浓缩;②培养1,3,5 d时,10 mmol/L Mg2++矿化胶原组细胞活力高于其余4组(P<0.05),矿化胶原组、5 mmol/L Mg2++矿化胶原组与空白对照组无差异(P>0.05),20 mmol/L Mg2++矿化胶原组低于空白对照组(P<0.05);③培养3 d后DAPI染色显示,20 mmol/L Mg2++矿化胶原组可见明显细胞核解体的现象,其余4组未见明显细胞核解体;④培养24 h后鬼笔环肽染色显示,除空白对照组、20 mmol/L Mg2++矿化胶原组外,其余3组细胞结构完整伸展,肌动蛋白微丝清晰明显,尤以10mmol/LMg2++矿化胶原组最为明显;⑤成骨诱导分化7 d后,除20 mmol/L Mg2++矿化胶原组外,其余3组碱性磷酸酶活性与Runx2基因表达均高于空白对照组(P<0.05),且10 mmol/L Mg2++矿化胶原组高于5 mmol/L Mg2++矿化胶原组、矿化胶原组(P<0.05);⑥结果表明,镁离子�
BACKGROUND:Preliminary study has shown that the composite materials composed of magnesium-based materials and mineralized collagen have a good supporting effect on repairing the critical defects,which can improve the mechanical strength of mineralized collagen and premature collapse during bone healing to some extent.However,magnesium-based metals degrade fast in chloride-containing solutions(including human body fluids or plasma),and the effects of releasing magnesium ions on the proliferation and differentiation of osteoblasts are unknown.OBJECTIVVE:To investigate the effects of magnesium ion combined with mineralized collagen on osteogenic differentiation of mouse preosteoblasts in vitro.METHODS:Mineralized collagen extracts were prepared from complete medium with magnesium ion concentration of 0,5,10,and 20 mmol/L.Mouse preosteoblasts were cultured with four mineralized collagen extracts,respectively,which were divided into mineralized collagen group,and 5,10 and 20 mmol/L Mg2++mineralized collagen groups.The mouse preosteoblasts cultured in complete medium were used as control group.The cell morphology,proliferation,apoptosis,intracellular microfilament actin,and the activity of alkaline phosphatase and expression level of the osteogenic gene Runx2 after osteogenic differentiation were detected.RESULTS AND CONCLUSION:(1)After 24 hours of culture,the cells in the mineralized collagen group,and 5 and 10 mmol/L Mg2++mineralized collagen groups adhered well,which showed no significant difference from the blank control group,and the elongated spindle cells with many synapses linked to the adjacent cells were observed.The cells in the 20 mmol/L Mg2++mineralized collagen group showed obvious pyknosis.(2)After 1,3 and 5 days of culture,the cell viability in the 10 mmol/L Mg2++mineralized collagen group was significantly higher than that in the other four groups(P<0.05).There was no significant difference among mineralized collagen,5 mmol/L Mg2++mineralized collagen and blank control groups(P>0.05).The cell viability
作者
孙溪饶
孙宝斋
张振保
Sun Xirao;Sun Baozhai;Zhang Zhenbao(2^nd Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,Liaoning Province,China;Graduate School of Jinzhou Medical University,Jinzhou 121000,Liaoning Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2020年第22期3467-3473,共7页
Chinese Journal of Tissue Engineering Research
基金
辽宁省自然科学基金计划重点项目(20180530071),项目参与者:孙溪饶、孙宝斋
辽宁省自然科学基金资助计划(2019-MS-141),项目负责人:孙溪饶。
关键词
矿化胶原人工骨
纳米羟基磷灰石/胶原
小鼠前成骨细胞
镁离子
矿化胶原
成骨细胞活力
骨修复材料
成骨分化
生物相容性
mineralized collagen artificial bone
nano-hydroxyapatite/collagen
mouse preosteoblasts
magnesium ions
mineralized collagen
viability of osteoblasts
bone repair materials
osteogenic differentiation
biocompatibility
