梯度糖溶液对体外培养成骨细胞影响的实验研究  

Effects of gradient glucose solution glucose on cultured osteoblasts in vitro

作　　者：苏友新[1] 陈智能[1] 杨连梓[2] 郑良朴[3] 

机构地区：[1]福建中医学院骨伤系,福建福州350003 [2]福建省第二人民医院 [3]福建中医学院中心实验室

出　　处：《中国骨伤》2005年第7期407-409,共3页China Journal of Orthopaedics and Traumatology

基　　金：福建省青年科技人才创新项目(编号:2001J062)

摘　　要：目的:探讨梯度高糖对体外培养成骨细胞(osteoblast,OB)的影响。方法:采用胰蛋白酶Ⅱ型胶原酶消化法从1～2日龄SD大鼠颅盖骨中分离出OB,倒置显微镜下及HE染色观察其形态,碱性磷酸酶(alkalinephosphatase,ALPase)染色法、钙化结节染色、VanGieson染色鉴定细胞后用不同浓度的糖溶液加入到成骨细胞培养体系中,作用不同时间,应用MTT比色法、ALPase含量测定、矿化结节形成量等来检测其对成骨细胞增殖和成骨能力的影响。结果:不同浓度的糖溶液在OB培养48h时具有明显的促进增殖作用,但>15mmol/L的溶液在72h时则呈抑制效应,其中以≤15mmol/L的糖溶液在培养早期有显著促进作用(P<0.05或P<0.01),而在培养较长时也无明显抑制效应;≥30mmol/L的糖溶液在OB培养7d时显著抑制ALPase的分泌(P<0.05);≥10mmol/L的糖溶液在培养18d时对矿化结节的形成也具有明显的延迟效应;<10mmol/L的糖溶液对OB分泌ALPase及矿化结节形成无抑制作用。结论:高糖可影响OB增殖、分化与矿化能力,OB增殖耐受的糖浓度为≤15mmol/L,OB分化与矿化过程耐受的糖浓度为<10mmol/L。Objective：To explore the effect of high gradient glucose on the proliferation and differentiation of osteoblastic cells（OB） cultured in vitro. Methods： OB was isolated from the skull of 1- 2 day newly born SD rats by means of Trypsin-collagenase digestion, and the osteoblasts were identified by morphologic analysis, VG collagen staining, ALPase staining, calcification nod staining etc. Different concentrations of glucose were added to the cultured OB. The effects of glucose on the proliferation and differentiation of osteoblasts was monitored by MTT analysis, ALPase activity analysis and formation of mineralized nodulus. Results：Different concentrations of glucose promoted the proliferation of osteoblastic cells for 48 hours culture, but inhibited for 72 hours when it was beyond 15 mmol/L. Among these different concentrations glucose, the glucose of less than 15 mmol/L could promote osteoblastic cells proliferation on the early period of cultivation （P 〈 0.01, P 〈 0.05） and has no suppression during a relative long time, and the glucose of more than 30 mmol/L inhibited the secretion of ALPase for 7 days（P 〈 0.05）, and the glucose of beyond 10 mmol/L could delay formation of mineralized cell nodulus for 18 days, below 10 mmol/L glucoses did not inhibit the secretion of ALPase and formation of mineralized nodulus. Conclusion：High gradient glucose could influence on the proliferation, differentiation and mineralization of osteoblastic cells. The tolerance concentrations of glucose were no more than 15 mmol/L on the proliferation of OB,and the differentiation and mineralization of OB were below 10 mmol/L.

关 键 词：骨形成 细胞培养 血糖 成骨细胞增殖 体外培养 糖溶液 梯度 实验研 ALPase mol/L 

分 类 号：R681[医药卫生—骨科学] R329-33[医药卫生—外科学]