Lozupone E, Palumbo C, Favia A, Ferretti M, Palazzini S, Cantatore F P
Institute of Human Anatomy, University of Bari, Italy.
Clin Rheumatol. 1996 Nov;15(6):563-72. doi: 10.1007/BF02238545.
The effect of mechanical stresses on osteogenesis, the viability of osteocytes and their metabolic activity in organ culture of bones intermittently loaded "in vitro" are reported. Metatarsal bones, isolated from 12-day-old rats, were cultured in BGJb medium (with 10% foetal calf serum, 75 micrograms/ml of ascorbic acid, 100 U/ml of penicillin and 100 micrograms/ml of streptomycin), in humidified air enriched by 5% CO2 and 30% O2, and loaded in our original device for 1/2 an hour at 1 Hz. homotypic isolated and unloaded bones, cultured in the same medium, were taken as controls. The ALP (alkaline phophatase activity) increases in the media of loaded bones in comparison with the control bones. The percentage of viable osteocytes is significantly greater in loaded than in control bones. TEM observations demonstrate that in both loaded and control unloaded bones, osteocytes show well developed organelle machinery and several gap junctions with adjacent cellular processes. In the cells of loaded bones, however, a higher number of cytoplasmic organelles and gap junctions were found. In particular, RER increases twice, gap junctions three times. The induced osteogenesis and the TEM observations demonstrate the suitability of this experimental model and support the recent advanced hypothesis according to which the mechanical loading may exert a trophic function on osteocytes, stimulating both the proteic synthesis in the above-mentioned cells and the cell-to-cell communication. Furthermore, the loading is likely to exert a biological stimulus on osteoblasts via signalling molecules produced by osteocytes.
本文报道了机械应力对“体外”间歇性加载的骨器官培养中骨生成、骨细胞活力及其代谢活性的影响。从12日龄大鼠分离出的跖骨,在含有10%胎牛血清、75微克/毫升抗坏血酸、100单位/毫升青霉素和100微克/毫升链霉素的BGJb培养基中,于含5%二氧化碳和30%氧气的湿润空气中培养,并在我们的原始装置中以1赫兹频率加载半小时。将在相同培养基中培养的同型分离且未加载的骨作为对照。与对照骨相比,加载骨培养基中的碱性磷酸酶(ALP)活性增加。加载骨中存活骨细胞的百分比显著高于对照骨。透射电镜观察表明,在加载骨和对照未加载骨中,骨细胞均显示出发育良好的细胞器结构以及与相邻细胞突起的多个间隙连接。然而,在加载骨的细胞中,发现细胞质细胞器和间隙连接的数量更多。特别是,粗面内质网增加了两倍,间隙连接增加了三倍。诱导的骨生成和透射电镜观察证明了该实验模型的适用性,并支持了最近提出的假说,即机械加载可能对骨细胞发挥营养功能,刺激上述细胞中的蛋白质合成以及细胞间通讯。此外,加载可能通过骨细胞产生的信号分子对成骨细胞施加生物刺激。