外源性NGF对体外培养的成骨细胞增殖、分化和矿化的影响
发布时间：2018-07-30 06:49
【摘要】： 目的 临床实践中常常会发现,截瘫和颅脑损伤患者伴随有四肢骨折时,往往在骨折断端可见到大量不成熟的骨痂生长,甚至肌肉内可以出现异位骨化。同时,一些神经疾病患者的骨折成骨量也较一般人多,因此可以认为神经系统参与了骨折的愈合过程。作为神经系统中重要的营养因子NGF对骨组织中最重要的细胞成骨细胞的影响现在还不是十分清楚。因此本实验研究外源性神经生长因子(NGF)对体外培养的成骨细胞增殖、分化和矿化的影响。 方法 取出生24h Wistar大鼠颅盖骨,剃净后剪成碎块,采用改良组织块混合酶消化法培养成骨细胞,将不同浓度的NGF(1pg%qml,10pg/ml,100pg/ml,500pg/ml,1000pg%qml)加入成骨细胞的培养基继续进行细胞培养,7天后用MTT法测定细胞的增殖情况,碱性磷酸酶试剂盒检测成骨细胞碱性磷酸酶的活性,免疫组织化学染色法检测Ⅰ型胶原的量,3周后茜素红钙染法检测矿化结节形成。 结果 对成骨细胞增殖的影响:低浓度的NGF(＜100 pg/ml)OD值明显较对照组高(P＜0.05),统计学上有显著性差异;对成骨细胞分化的影响:500 pg/ml,100 pg/ml,10 pg/ml这三个浓度组的ALP活性(OD值)较对照组高(P＜0.05),分泌Ⅰ型胶原的量(OD值)也较其他各组高(P＜0.05),差异有统计学意义;对成骨细胞矿化的影响,各组无显著性差异。 结论 外源性NGF对体外培养的成骨细胞具有增殖作用、可以促进成骨细胞的分化,增强ALP的活性,促进Ⅰ型胶原的分泌,但不能明显的促进成骨细胞的矿化。
[Abstract]:Objective it is often found in clinical practice that patients with paraplegia and craniocerebral injury accompanied by limb fracture often see a large number of immature callus growth and even heterotopic ossification in the muscle. At the same time, some patients with neurological diseases have more bone formation, so the nervous system can be considered to be involved in the process of fracture healing. The effects of NGF, an important nutritional factor in the nervous system, on osteoblasts, which are the most important in bone tissue, are not well understood. Therefore, the effects of exogenous nerve growth factor (NGF) on proliferation, differentiation and mineralization of osteoblasts in vitro were studied. Methods the skulls of 24 hours old Wistar rats were taken out and cut into pieces after shaving. Osteoblasts were cultured by modified tissue mass mixed enzyme digestion method. Adding different concentrations of NGF (10pg / ml 10pg / ml) to the culture medium of osteoblasts (500pg / ml) for 7 days, the proliferation of osteoblasts was determined by MTT assay, and the activity of alkaline phosphatase (ALP) was detected by alkaline phosphatase assay. Immunohistochemical staining was used to detect the amount of type I collagen. 3 weeks later alizarin red calcium staining was used to detect the formation of mineralized nodules. Results the effect on osteoblast proliferation: the OD value of low concentration NGF (< 100 pg/ml) was significantly higher than that of control group (P < 0. 05). The effect on the differentiation of osteoblasts in the three concentration groups: 500pg / ml ~ 100pg / ml ~ (10 pg/ml) was higher than that of the control group (P < 0. 05), and the OD value of the three concentration groups was also higher than that of the other groups (P < 0. 05), and the effect on the mineralization of osteoblasts was also significant (P < 0. 05). There was no significant difference in each group. Conclusion exogenous NGF can promote the differentiation of osteoblasts, enhance the activity of ALP and promote the secretion of type I collagen, but it can not obviously promote the mineralization of osteoblasts.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329.2
本文编号：2154184
[Abstract]:Objective it is often found in clinical practice that patients with paraplegia and craniocerebral injury accompanied by limb fracture often see a large number of immature callus growth and even heterotopic ossification in the muscle. At the same time, some patients with neurological diseases have more bone formation, so the nervous system can be considered to be involved in the process of fracture healing. The effects of NGF, an important nutritional factor in the nervous system, on osteoblasts, which are the most important in bone tissue, are not well understood. Therefore, the effects of exogenous nerve growth factor (NGF) on proliferation, differentiation and mineralization of osteoblasts in vitro were studied. Methods the skulls of 24 hours old Wistar rats were taken out and cut into pieces after shaving. Osteoblasts were cultured by modified tissue mass mixed enzyme digestion method. Adding different concentrations of NGF (10pg / ml 10pg / ml) to the culture medium of osteoblasts (500pg / ml) for 7 days, the proliferation of osteoblasts was determined by MTT assay, and the activity of alkaline phosphatase (ALP) was detected by alkaline phosphatase assay. Immunohistochemical staining was used to detect the amount of type I collagen. 3 weeks later alizarin red calcium staining was used to detect the formation of mineralized nodules. Results the effect on osteoblast proliferation: the OD value of low concentration NGF (< 100 pg/ml) was significantly higher than that of control group (P < 0. 05). The effect on the differentiation of osteoblasts in the three concentration groups: 500pg / ml ~ 100pg / ml ~ (10 pg/ml) was higher than that of the control group (P < 0. 05), and the OD value of the three concentration groups was also higher than that of the other groups (P < 0. 05), and the effect on the mineralization of osteoblasts was also significant (P < 0. 05). There was no significant difference in each group. Conclusion exogenous NGF can promote the differentiation of osteoblasts, enhance the activity of ALP and promote the secretion of type I collagen, but it can not obviously promote the mineralization of osteoblasts.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329.2
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相关期刊论文 前3条
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