氟化钠对人黄韧带细胞碱性磷酸酶活性及骨钙素合成的影响

目的 探讨氟化钠(NaF)对体外培养条件下人黄韧带细胞碱性磷酸酶(alkaline phosphatase,ALP)活性及骨钙素(bone gla protein,BGP)合成的影响.方法 依据手术标本来源不同,分为正常黄韧带细胞(NLF)组(取自急性外伤性胸腰椎骨折截瘫患者,7例)、退变黄韧带细胞(DLF)组(取自退变性腰椎管狭窄症患者,9例)及骨化黄韧带细胞(OLF)组(取自胸椎黄韧带骨化患者,8例).采用组织块贴壁法进行体外细胞培养,共获得24组传代细胞.取第五代细胞加入不同浓度的NaF,观察细胞形态变化,并测定NaF对各组人黄韧带细胞的ALP活性和BGP合成的影响.结果 不同来源的人黄韧带细胞均可在体外增殖并传代,DLF与OLF组人黄韧带细胞呈多形性表现,并可形成钙结节.体外培养条件下,高浓度(1.0mmol/L)NaF可导致人黄韧带细胞发生中毒反应；低浓度(0.01～0.125 mmol/L)NaF可促进DLF组黄韧带细胞增殖、钙结节形成,ALP活性上调,BGP合成明显增加,而对OLF与NLF组人黄韧带细胞则无明显效应.结论 体外培养条件下,低浓度(0.01～0.125 mmol/L)NaF可促进人退变黄韧带细胞的ALP活性增高及BGP合成增加,提示低浓度NaF可能促进人退变黄韧带细胞向成骨方向分化.

Influence of sodium fluoride on alkaline phosphatase activity and bone gla protein synthesis in human yellow ligament cells in vitro

Objective To investigate the influence of sodium fluoride(NaF)on alkaline phosphatase (ALP)activity and bone gla protein(BGP)synthesis in yellow ligament cells from different surgical simples in vitro.Methods The human ligament cells were divided into three groups according to its sources,including normal yellow ligament cells(NLF)group(from acute traumatic thoracolumbar fractures with paraplegia in 7 patients),degenerative yellow ligament cells(DLF)group(from degenerative lumbar stenosis in 9 cases)and ossified ligament cells(OLF)group(thoracic yellow ligament from 8 patients).Twenty-four groups of cells were obtained under vitro cell culture by the method of tissue adherence.Different concentrations of NaF were added into the medium when the cells spread to the fifth generation.Then,the morphological changes were observed and ALP activity and BGP synthesis were tested.Results Human yellow ligament cells from different samples can proliferate and be passaged in vitro.The cell in ossific groups and degenerative groups were pleomorphic and could form calcium nodules.High concentration of NaF(1.0 mmol/L)can lead to cytotoxic reaction in all 24 groups.Low concentrations of(0.01-0.125 mmol/L)NaF can enhance the ALP activity and BGP synthesis in DLF groups while no effect was found in OLF and NLF groups cells under the same concentration of NaF.Conclusion The fact that fluoride can promote ALP activity and BGP synthesis in degenerative yellow ligament cells in vitro indicates fluoride may play an important role in inducing further ossification of human ligament cells.