不同成骨诱导体系对MC3T3-E1成骨分化的影响

doi:10.13701/j.cnki.kqyxyj.2024.04.005

摘要/Abstract

摘要： 目的: 明确前成骨细胞MC3T3-E1最佳体外成骨诱导体系。方法: 构建不同成骨诱导体系,分别为A组地塞米松0.1 μmol/L、抗坏血酸50 μg/mL及β-甘油磷酸钠10 mmol/L;B组地塞米松0.01 μmol/L、抗坏血酸50 μg/mL及β-甘油磷酸钠10 mmol/L;C组地塞米松1 μmol/L、抗坏血酸50 μg/mL及β-甘油磷酸钠10 mmol/L;D组抗坏血酸50 μg/mL及β-甘油磷酸钠10 mmol/L;E组抗坏血酸100 μg/mL及β-甘油磷酸钠5 mmol/L。对MC3T3-E1进行为期7 d、14 d的体外成骨诱导,通过碱性磷酸酶染色、茜素红染色对在不同成骨诱导体系作用下MC3T3-E1成骨分化的影响进行分析,通过RT-qPCR检测成骨分化相关基因ALP、Runx2、COL-1、OCN、OPN表达水平,比较不同成骨诱导体系的诱导性能。结果: B、D和E组茜素红定量最优,差异具有统计学意义(P＜0.05),诱导7 d的RT-qPCR结果显示C组ALP、Runx2、COL-1的基因表达量最高(P＜0.05)。诱导14 d的RT-qPCR结果显示:OCND组表达最高(P＜0.05)。结论: 地塞米松和抗坏血酸的浓度均对MC3T3-E1的成骨分化产生影响,C组1 μmol/L地塞米松、50 μg/mL抗坏血酸与10 mmol/L β-甘油磷酸钠早期成骨更佳,选择其作为早期成骨验证,D组50 μg/mL抗坏血酸与10 mmol/L β-甘油磷酸钠晚期成骨更佳,但综合考虑,建议B组0.01 μmol/L地塞米松、50 μg/mL抗坏血酸与10 mmol/L β-甘油磷酸钠成骨诱导体系用于MC3T3-E1体外晚期成骨研究。

关键词: MC3T3-E1, 成骨分化, 诱导体系

Abstract: Objective: To determine the optimal osteogenic induction system in vitro for MC3T3-E1 osteoblasts. Methods: Different osteogenic induction systems were constructed, which were dexamethasone 0.1 μmol/L, ascorbic acid 50 μg/mL, and β-sodium glycerophosphate 10 mmol/L in group A; dexamethasone 0.01 μmol/L, ascorbic acid 50 μg/mL, and β-sodium glycerophosphate 10 mmol/L in group B; dexamethasone 1 μmol/L, ascorbic acid 50 μg/mL, and β-sodium glycerophosphate 10 mmol/L in group C; ascorbic acid 50 μg/mL and β-sodium glycerophosphate 10 mmol/L in Group D; and ascorbic acid 100 μg/mL and β-sodium glycerophosphate 5 mmol/L in Group E. MC3T3-E1 cells were induced in vitro for 7 days and 14 days. The effects of alkaline phosphatase staining and alizarin red staining on osteogenic differentiation of MC3T3-E1 were analyzed. The expression levels of osteogenic differentiation related genes (ALP, Runx-2, COL-1, OCN, OPN) were detected by RT-qPCR. Results: There was a significant difference in the optimal quantification of alizarin among groups B, D, and E (P<0.05). The PCR results at 7 days after induction showed that the gene expression levels of ALP, Runx-2, and COL-1 in group C were the highest (P<0.05). PCR results at 14 days after induction showed that the expression of OCN in Group D was the highest (P<0.05). Conclusion: The concentrations of dexamethasone and ascorbic acid have an effect on the osteogenic differentiation of MC3T3-E1 cells. Group C was better for early osteogenesis and Group D was better for late osteogenesis. However, in comprehensive consideration, Group B is recommended for late osteogenesis.

Key words: MC3T3-E1, osteogenic differentiation, inducement system

管馨, 陈安琪, 赖颖真, 吴勇敏, 翁鑫泽, 王伟新, 雷奕宣. 不同成骨诱导体系对MC3T3-E1成骨分化的影响[J]. 口腔医学研究, 2024, 40(4): 304-309.

GUAN Xin, CHEN Anqi, LAI Yingzhen, WU Yongmin, WENG Xinze, WANG Weixin, LEI Yixuan. Effects of Different Osteogenic Induction Systems on Osteogenic Differentiation of MC3T3-E1 Cells[J]. Journal of Oral Science Research, 2024, 40(4): 304-309.

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