REACTIVATION OF INHIBITED BONE ACID-PHOSPHATASE AND ITS SIGNIFICANCE IN BONE HISTOMORPHOMETRY
REACTIVATION OF INHIBITED BONE ACID-PHOSPHATASE AND ITS SIGNIFICANCE IN BONE HISTOMORPHOMETRY
复制标题
DOI：
10.1177/35.12.3680930
复制
发表时间：
1987-12-01
影响因子：
3.2
通讯作者：
HOWARD, GA
中科院分区：
文献类型：
--
作者：
LIU, CC;SHERRARD, DJ;HOWARD, GA
尽管在骨中有酸性磷酸酶（AcP）激活或再激活的生化证明，但很少有人尝试在组织化学上显示类似的效果。生长中的大鼠骨骼，在室温下固定在4%缓冲甲醛中，在5%甲酸中脱矿，显示出预期的AcP失活。然而，被抑制的AcP在37度下预孵育1小时后被重新激活。C在以下缓冲液中：0.2 M Tris， 0.2 M甘氨酸，0.2 M NaHCO3，或0.1 M硼砂，以及在碱性水中，但不在0.2 M Na2HOP4中（pH均为9）。这种再激活是(a)位点特异性的（例如，破骨细胞、成骨细胞、骨细胞和水泥系），(b)温度和ph依赖性的，(c)不受OH-或SH-结合剂或碱性磷酸酶抑制剂的影响，(d)被10 mM Na2HPO4完全抑制。与先前报道的方法相比，该再激活过程更简单和/或更有效，研究人员在83块活组织活检骨中观察到，这些活组织活检骨包埋在甲基丙烯酸甲酯中，并在冷缓冲甲醛中储存了7个月。本研究展示了一种独特的方法来重新激活和定位骨中被抑制的AcP，并提出了在骨组织形态学测量中的可能应用。
Despite biochemical demonstration of acid phosphatase (AcP) activation or reactivation in bone, few attempts have been made to show similar effects histochemically. Bones from growing rats, when fixed in 4% buffered formaldehyde at room temperature and demineralized in 5% formic acid, exhibited expected inactivation of AcP. The inhibited AcP, however, was reactivated by pre-incubation of sections for 1 hr at 37.degree. C in the following buffers: 0.2 M Tris, 0.2 M glycine, 0.2 M NaHCO3, or 0.1 M borax, as well as in alkaline water, but not in 0.2 M Na2HOP4 (all at pH 9). The reactivation was (a) site-specific (e.g., osteoclasts, osteoblasts, osteocytes, and cement lines), (b) temperature- and pH-dependent, (c) unaffected by OH- or SH--binding agents or by an alkaline phosphatase inhibitor, and (d) inhibited completely by 10 mM Na2HPO4. The reactivation process, much simplified and/or more effective than with the methods previously reported, was observed in all 83 human biopsy bones embedded in methyl methacrylate and in human bones stored in cold buffered formaldehyde for 7 months. This study demonstrates a unique method for reactivating and thus localizing the inhibited AcP in bones, and suggests possible applications in bone histomorphometry.