Shibutani T, Heersche J N
MRC Group in Periodontal Physiology, Faculty of Dentistry, University of Toronto, Ontario, Canada.
J Bone Miner Res. 1993 Mar;8(3):331-6. doi: 10.1002/jbmr.5650080310.
We investigated the effect of medium pH on activity of isolated osteoclasts and have also looked at the possibility that medium pH affects osteoclast numbers during culture. Osteoclast-containing cell suspensions prepared from neonatal rabbits were cultured on bovine bone slices at pH 6.5, 7.0, or 7.5. After 24 or 48 h of culture, the cells and bone slices were fixed and stained for tartrate-resistant acid phosphatase (TRAP). After counting the osteoclasts, the cells were removed and the resorption lacunae stained by immunostaining using anticollagen type I antibody and then quantitated. We found that the resorptive activity of isolated rabbit osteoclasts was sharply increased at pH 6.5-7. Osteoclast differentiation and proliferation, on the other hand, were optimal at pH 7.0-7.5 but decreased at pH 6.5. The results thus imply that pH regulation of the bone surface environment can dramatically alter both the number of osteoclasts and their resorptive activity.
我们研究了培养基pH值对分离的破骨细胞活性的影响，并且还探讨了培养基pH值在培养过程中影响破骨细胞数量的可能性。从新生兔制备的含破骨细胞的细胞悬液在pH值为6.5、7.0或7.5的条件下，接种于牛骨切片上进行培养。培养24或48小时后，将细胞和骨切片固定，并进行抗酒石酸酸性磷酸酶（TRAP）染色。在对破骨细胞计数后，去除细胞，然后用抗I型胶原抗体进行免疫染色，对吸收陷窝进行染色并定量。我们发现，分离的兔破骨细胞的吸收活性在pH值为6.5 - 7时急剧增加。另一方面，破骨细胞的分化和增殖在pH值为7.0 - 7.5时最佳，但在pH值为6.5时降低。因此，结果表明骨表面环境的pH调节可显著改变破骨细胞的数量及其吸收活性。
