摘要
目的研究地塞米松对体外培养的成人成骨细胞增殖和分化的影响。方法取成人的髂骨松质骨,采用胶原-胰蛋白酶消化法,获得松质骨中的成骨细胞,进行纯化和培养。在此基础上,添加1×10-8mol/L的地塞米松,连续加药3d,置入CO2孵箱中培养,同时作不加药的空白对照。通过倒置相差显微镜和电镜观察增殖和分化情况,检测上清液中的碱性磷酸酶和骨钙素含量,并且对成骨细胞的凋亡情况进行测定。结果胶原-胰蛋白酶消化法得到的成人成骨细胞,生长情况良好,生化指标稳定可靠,细胞纯化效果佳,可以满足相关研究的需要。进一步研究结果显示,在地塞米松浓度为1×10-8mol/L,成骨细胞密度为10000/ml的条件下,上清液中碱性磷酸酶和骨钙素含量明显增高,骨结节形成加速,与未加药的对照组比较差异均有非常显著性意义,同时成骨细胞的凋亡情况无明显变化。结论地塞米松对体外培养的成人成骨细胞的增殖和分化有明显地促进作用,可以作为促进骨形成的阳性对照应用于抗骨质疏松药物的筛选研究。
Objective To observe and analyze the effect of dexamethasone on the adult osteoblast proliferation and differentiation in vitro. Methods Iliac trabecular bone specimens were obtained from eight adult patients undergoing scheduled surgery. After the bone pieces were treated with collagenase trypsin, larage numbers of osteoblasts were released out. Then the osteoblasts were purified and their passages were fed with DMED-F12 and fetal bovine serum. The cells were incubated under the condition of 37 ℃, 95% relative humidity and 5% CO2. Subsequently, 1×10-8 mol/L dexamethasone were added into culture medium to feed eight groups of osteoblasts for three days and the control groups were cultured not to add medicine at the same time. All the groups were observed continually under light microscope and transmission electron microscope. Finally, apoptosis detection was performed by use of the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and biochemical indexes alkaline phosphatase (ALP) and osteocalcin (OCN), were assayed to check if dexamethasone has any effect on proliferation and differentiation of osteoblasts in vitro. Results Of the adult osteoblasts obtained by collagenase trypsin method, the survival condition was good, the biochemical indexes were stable, the degree of purification was excellent, and they were fit for the next step study. The following data demonstrated that under the condition of dexamethasone 1×10-8 mol/L and osteoblasts 10 000 /ml, there were significant increase of ALP and OCN secretion without apoptosis alteration. Conclusion Dexamethasone has significant promoting effect on proliferation and differentiation of adult osteoblasts in vitro without apoptosis alteration and could be used as positive control during screening research of osteoporosis.
出处
《中华骨科杂志》
CAS
CSCD
北大核心
2001年第8期493-497,共5页
Chinese Journal of Orthopaedics
关键词
成骨细胞
细胞培养
地塞米松
细胞增殖
细胞分化
Osteoblasts
Cell culture
Dexamethasone
Proliferation
Cell differentiation
