Fe~(3+)和Fe~(2+)对原代培养的成骨细胞增殖、分化和矿化功能的影响

摘要

采用噻唑蓝(MTT)法、碱性磷酸酶(ALP)比活性测定、油红O染色和茜素红染色及定量分析,研究了不同浓度的Fe~(3+)和Fe~(2+)对原代培养的成骨细胞增殖、分化及矿化功能的影响.结果表明:浓度为1×10~(-9)～1×10~(-4) mol·L~(-1)的Fe~(3+)和Fe~(2+)促进成骨细胞增殖,但是在较高浓度1×10~(-3) mol·L~(-1)时,它们则抑制成骨细胞增殖.与成骨细胞作用48 h,浓度为1×10~(-8)～1×10~(-4) mol·L~(-1)的Fe~(3+)和Fe~(2+)抑制其分化,但在较低的浓度1×10~(-9) mol·L~(-1)时则对其分化没有影响:进一步延长作用时间为72 h,Fe~(3+)对成骨细胞分化没有影响,除1×10~(-6)mol·L~(-1)浓度的Fe~(2+)促进成骨细胞分化外,其他浓度的Fe~(2+)则抑制其分化;测试浓度下的Fe~(3+)对成骨细胞向脂肪细胞的横向分化表现为抑制或没有影响,而Fe~(2+)的影响则依赖于浓度和作用时间.在1×10~(-8)～1×10~(-5)mol·L~(-1)浓度范围内,Fe~(3+)和Fe~(2+)对矿化结节的影响表现出相反的效应.在较高浓度(1×10~(-4)mol·L~(-1))下,它们促进矿化节结的形成,而在较低浓度(1×10~(-9)mol·L~(-1))下,Fe~(3+)抑制矿化节结的形成,Fe~(2+)则没有影响.结果提示:浓度.作用时间和铁离子的价态都是影响Fe~(3+)和Fe~(2+)生物效应(从毒性到活性,从损伤到保护,从上调到下调)转变的关键因素.%The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromid (MTT), alkaline phosphatase (ALP) activity, oil red O assays and alizarin red-S(ARS) stain were used to evaluate the effects of Fe~(3+) and Fe~(2+) on proliferation, differentiation and mineralization function of primary osteoblasts (OBs) in vitro. The results indicate that both Fe~(3+) and Fe~(2+)(1×10~(-9)～1×10~(-4) mol·L~(-1)) promote the proliferation of OBs, but turn to inhibit at a higher concentration of 1×10~(-3) mol·L~(-1). Both Fe~(3+) and Fe~(2+)(1×10~(-8)～1×10~(-4) mol·L~(-1)) inhibit differentiation of OBs, but have no effect on differentiation at a lower concentration of 1×10~(-9) mol·L~(-1) for 48 h. Whereas, Fe~(3+) shows no effect on differentiation of OBs, Fe~(2+) begins to promote differentiation of OBs at concentration of 1×10~(-6) mol·L~(-1), but Fe~(2+) inhibits differentiation of OBs at other concentrations by prolonging the incubation time. Fe~(3+) inhibits adipocytic trans-differentiation of OBs or has no effects on adipocytic trans-differentiation of OBs at tested concentrations. The promotion or inhibition effect of Fe~(2+) on adipocytic trans-differentiation of OBs depends on concentration and incubation time. Fe~(3+) and Fe~(2+)(1×10~(-8)～1 ×10~(-5) mol·L~(-1)) inhibit and promote the formation of mineralized matrix nodules of OBs, respectively. Fe~(3+) and Fe~(2+) promote the formation of mineralized matrix nodules at a higher concentration of 1×10~(-4) mol· L~(-1), but Fe~(3+) inhibits the formation of mineralized matrix nodules, Fe~(2+) shows no effect at a lower concentration of 1×10~(-9) mol·L~(-1). The results suggest that concentration, culture time and valence state of iron ion are key factors for switching the biological effects of Fe~(3+) and Fe~(2+) from toxicity to activity, from damage to protection, or from down-regulation to up-regulation.