Wang Qiu-shi, Pei Guo-xian, Jinag Shan, Zhao Pei-ran, Liang Shuang-wu, Dai Jin-liang, Cui Jian-de
Department of Orthopedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2008 May;28(5):774-8.
To investigate the effect of rabbit saphenous and sciatic nerve homogenates on the proliferation and calcification of rabbit osteoblasts in vitro.
The saphenous nerves (sensory nerves) and the muscular branches of the sciatic nerve (motor nerve) were collected from 48 New Zealand white rabbits to prepare the nerve tissue homogenates. Bone marrow mesenchymal stem cells (MSCs) were isolated from the rabbits and cultured in vitro, and after 14 days of routine osteogenic induction, the resultant osteoblasts were identified by immunohistochemistry, alkaline phosphatase (ALP) and Alizarin red S staining. The osteoblasts were then incubated in the induction medium containing the saphenous (sensory nerve group) or sciatic homogenates (motor nerve group), with the cells in the dexamethasone-containing, dexamethasone-free osteogenic induction medium and control medium as the control. The proliferation, total protein and ALP activity of the osteoblasts were measured every other day until the 8th day, and Alizarin red S staining was used for quantitative analysis of calcification of the cells after two weeks.
The application of the saphenous nerve homogenates significantly promoted cell proliferation, total protein and ALP activity (P<0.01, P<0.05 and P<0.05), while exposure of the osteoblasts to dexamethasone inhibited the cell proliferation (P<0.001). Compared to dexamethasone-free group, the saphenous homogenates enhanced the mineralization of the osteoblasts (P<0.001).
Saphenous nerve homogenates significantly promotes the proliferation, differentiation, ALP activity and mineralization of rabbit osteoblasts, but sciatic nerve homogenates do not show osteogenic effects on the cells.
研究兔隐神经和坐骨神经匀浆对兔成骨细胞体外增殖及钙化的影响。
从48只新西兰白兔中采集隐神经(感觉神经)和坐骨神经的肌支(运动神经)制备神经组织匀浆。从兔体内分离骨髓间充质干细胞(MSCs)并进行体外培养,常规成骨诱导14天后,通过免疫组织化学、碱性磷酸酶(ALP)和茜素红S染色鉴定所得成骨细胞。然后将成骨细胞接种于含隐神经匀浆的诱导培养基(感觉神经组)或坐骨神经匀浆的诱导培养基(运动神经组)中培养,以含地塞米松、不含地塞米松的成骨诱导培养基及对照培养基中的细胞作为对照。每隔一天检测成骨细胞的增殖、总蛋白及ALP活性,直至第8天,并在两周后用茜素红S染色对细胞钙化进行定量分析。
隐神经匀浆的应用显著促进细胞增殖、总蛋白及ALP活性(P<0.01、P<0.05和P<0.05),而成骨细胞暴露于地塞米松中则抑制细胞增殖(P<0.001)。与不含地塞米松组相比,隐神经匀浆增强了成骨细胞的矿化作用(P<0.001)。
隐神经匀浆显著促进兔成骨细胞的增殖、分化、ALP活性及矿化,但坐骨神经匀浆对细胞未显示有成骨作用。