外源性NGF對體外培養(yǎng)的成骨細胞增殖、分化和礦化的影響
發(fā)布時間：2018-07-30 06:49
【摘要】： 目的 臨床實踐中常常會發(fā)現(xiàn),截癱和顱腦損傷患者伴隨有四肢骨折時,往往在骨折斷端可見到大量不成熟的骨痂生長,甚至肌肉內(nèi)可以出現(xiàn)異位骨化。同時,一些神經(jīng)疾病患者的骨折成骨量也較一般人多,因此可以認為神經(jīng)系統(tǒng)參與了骨折的愈合過程。作為神經(jīng)系統(tǒng)中重要的營養(yǎng)因子NGF對骨組織中最重要的細胞成骨細胞的影響現(xiàn)在還不是十分清楚。因此本實驗研究外源性神經(jīng)生長因子(NGF)對體外培養(yǎng)的成骨細胞增殖、分化和礦化的影響。 方法 取出生24h Wistar大鼠顱蓋骨,剃凈后剪成碎塊,采用改良組織塊混合酶消化法培養(yǎng)成骨細胞,將不同濃度的NGF(1pg%qml,10pg/ml,100pg/ml,500pg/ml,1000pg%qml)加入成骨細胞的培養(yǎng)基繼續(xù)進行細胞培養(yǎng),7天后用MTT法測定細胞的增殖情況,堿性磷酸酶試劑盒檢測成骨細胞堿性磷酸酶的活性,免疫組織化學染色法檢測Ⅰ型膠原的量,3周后茜素紅鈣染法檢測礦化結節(jié)形成。 結果 對成骨細胞增殖的影響:低濃度的NGF(＜100 pg/ml)OD值明顯較對照組高(P＜0.05),統(tǒng)計學上有顯著性差異;對成骨細胞分化的影響:500 pg/ml,100 pg/ml,10 pg/ml這三個濃度組的ALP活性(OD值)較對照組高(P＜0.05),分泌Ⅰ型膠原的量(OD值)也較其他各組高(P＜0.05),差異有統(tǒng)計學意義;對成骨細胞礦化的影響,各組無顯著性差異。 結論 外源性NGF對體外培養(yǎng)的成骨細胞具有增殖作用、可以促進成骨細胞的分化,增強ALP的活性,促進Ⅰ型膠原的分泌,但不能明顯的促進成骨細胞的礦化。
[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.
【學位授予單位】：中國醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R329.2
本文編號：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.
【學位授予單位】：中國醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R329.2
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相關期刊論文 前3條
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2 馬昕;黃鋼勇;夏新雷;金慰芳;姜建元;黃煌淵;;大鼠腦內(nèi)成分對體外培養(yǎng)的成骨細胞增殖、分化和礦化的影響[J];復旦學報(醫(yī)學版);2006年03期
3 周赤兵;張遠金;蔡冬冰;段軍;;局部運用神經(jīng)生長因子促進骨折愈合的實驗研究[J];中國傷殘醫(yī)學;2007年06期
，本文編號：2154184
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