建模时间及糖浓度对糖尿病大鼠下颌骨成骨细胞培养的影响

Effect of diabetic duration and glucose concentration on the culture of osteoblasts obtained from diabetic rats’ mandibles

目的:探讨建模时间长短及不同糖浓度培养基对糖尿病大鼠下颌骨来源成骨细胞培养的影响为胰岛素经人工种植牙给药建立细胞模型。方法:链脲霉素诱导Ⅰ型糖尿病大鼠模型,建模后10d及40d取其下颌骨,分别于葡萄糖含量为5.5mmol/L(低糖组)和25mmol/L(高糖组)的培养基中进行成骨细胞培养,观察细胞的形态及生长情况。细胞计数法检测生长增殖能力,碱性磷酸酶染色法、茜素红钙结节染色法分别检测分化及矿化能力。结果:细胞增殖能力由大到小依次为建模10d低糖组>建模10d高糖组>建模40d低糖组>建模40d高糖组(P<0.05),分化能力建模10d低糖组>建模40d低糖组、建模10d高糖组及建模40d高糖组(P<0.01)。建模10d低糖组可形成钙结节,其他组未能形成钙结节。结论:建模时间延长、高糖培养对糖尿病大鼠下颌骨来源成骨细胞的各项生物学活性指标均产生负面影响,其中建模时间主要抑制了细胞增殖,而高糖培养主要抑制了细胞的分化和矿化。

Objective： To investigate the effect of diabetic duration and different glucose concentration on the culture of rat mandibular osteoblast , and establish cell model for insulin delivery via dental implants, Methods： The mandibles harvested from type I diabetic rats induced by streptozotocin were used for the primary culture. All the mandibles were divided by the diabetic duration time （10d or 40d） and glucose concentration of DMEM（5.5 or 25mmol/L）.Comparison of cell biological activity was made among the four groups with cell count, alkaline phosphorase stain and alizarin bordeaux stain of calcified nodules were performed to lest the differentiation and mineralization. Results： The proliferation ability, from high to low, was 10d low glucose group〉 10d high glucose group〉 40d low glucose group〉 40d high glucose group （P〈0.05）. The differentiation ability, from high to low, was 10d low glucose group〉 40d low glucose group and high glucose groups（P 〈0.01）. Alizarin bordeaux stain of calcified nodules was positive in 10d low glucose group, but negative in other groups. Conclusion ： The osteoblasts＇ biological activity was downregulated by long duration of diabetes mellitus and hyperglycemia. And long duration of diabetes mellitus mainly affects cell proliferation, yet hyperglycemia mainly affects cell differentiation and mineralization.