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| 乳酸对成骨样细胞成骨表型及碱性磷酸酶基因表达的影响研究 |
| Effects of lactic acid on osteoblastic phenotype and alkaline phosphatase gene expression of osteoblast-like cells |
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| DOI：10.3969/j.issn.1006-7108.2023.08.002 |
| 中文关键词: 聚乳酸 乳酸 成骨样细胞 成骨表型 基因表达 |
| 英文关键词:polylactic acid lactic acid osteoblast-like cells osteoblastic phenotype gene expression |
| 基金项目:国家自然科学基金项目（30800223）；山西省基础研究计划项目(20210302124313)；山西省高等学校科技创新计划项目(2021L543)；太原工业学院引进人才科研项目(2022KJ044) |
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| 中文摘要: |
| 目的 明确聚乳酸降解终产物乳酸对细胞成骨表型和基因表达的影响机理，为聚乳酸支架的精确设计提供参考意义。方法 配制乳酸浓度为8、16、22、27 mmol/L的培养基，将不含乳酸培养基作为对照组，检测培养基的pH和渗透压。利用培养基培养大鼠成骨样细胞Ros17/2.8，观察细胞形态，测定细胞I型胶原、碱性磷酸酶（alkaline phosphatase，ALP）活性和骨钙素含量，盐酸四环素染色钙结节并定量分析。采用实时定量PCR法检测细胞ALP mRNA表达。结果 与对照组相比，随着乳酸浓度增加，培养基pH下降，渗透压升高，成骨样细胞形态改变，数量减少。低浓度乳酸组（8、16 mmol/L）的I型胶原量都显著高于对照组，其余组与对照组相比无显著差异。含乳酸组的细胞ALP活性都低于对照组，并且乳酸浓度越高，细胞ALP活性越低。含乳酸组的骨钙素量与对照组之间没有显著差异。随着乳酸浓度增加，含乳酸组细胞所形成的钙结节阳性面积减少，都低于对照组。低浓度乳酸组（8、16 mmol/L）的细胞ALP mRNA表达量都低于对照组。结论 聚乳酸降解终产物乳酸可抑制细胞ALP mRNA的表达，降低ALP活性。随着乳酸浓度增加，乳酸抑制作用增强，最终降低细胞的成骨矿化能力。此外，乳酸不会抑制细胞分泌I型胶原和骨钙素。 |
| 英文摘要: |
| Objective It will provide references for precise design of PLA scaffolds to clarify the effect of final degradation products of polylactic acid (PLA) on osteoblastic phenotype and gene expression of cells. Methods The medium with lactic acid in concentrations of 8, 16, 22, 27 mmol/L were prepared and the medium without lactic acid was as control group. The pH and osmolality of the medium were detected. The rat osteoblast-like cells Ros17/2.8 was cultured with medium containing lactic acid (8, 16, 22, 27 mmol/L). Cell morphology was observed. The type I collagen, alkaline phosphatase (ALP) activity and osteocalcin content of cells were determined. The calcium nodules were stained with tetracycline hydrochloride and quantitatively analyzed. Real-time quantitative PCR method was used to detect ALP mRNA expression level of cells. Results Compared with control group, the pH of medium was decreased with increase of concentration of lactic acid while the osmolality was increased. The morphology of osteoblast-like cells was changed and the number of cells was decreased. The amount of type I collagen in medium containing low-concentration lactic acid (8, 16 mmol/L) was significantly higher than that of control group, and there was no significant difference between the other groups and control group. With the increase of concentration of lactic acid in medium, the ALP activity of cells was decreased compared with control group. There was no significant difference between medium containing lactic acid groups and control group for the amount of osteocalcin. As the concentration of lactic acid in medium increases, the positive area of calcium nodules was decreased compared with control group. The expression level of ALP mRNA was lower than that of control group in medium containing low-concentration lactic acid (8, 16 mmol/L). Conclusion Lactic acid was the final degradation products of PLA. The expression level of ALP mRNA was inhibited and the ALP activity of cells was decrease by lactic acid. The inhibitory effect of lactic acid in ALP activity was increased with the increase of lactic acid concentration in medium. The mineralization of osteoblast-like cells was reduced by lactic acid finally. In addition, the secretion of type I collagen and osteocalcin of cells was not inhibited by lactic acid. |
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