Boskey Adele L, Gelb Bruce D, Pourmand Eric, Kudrashov Valery, Doty Stephen B, Spevak Lyudmila, Schaffler Mitchell B
Musculoskeletal Integrity Program, Hospital for Special Surgery, New York, NY 10021, USA.
Calcif Tissue Int. 2009 Mar;84(3):229-39. doi: 10.1007/s00223-008-9214-6. Epub 2009 Jan 27.
Cathepsin K deficiency in humans causes pycnodysostosis, which is characterized by dwarfism and osteosclerosis. Earlier studies of 10-week-old male cathepsin K-deficient (knockout, KO) mice showed their bones were mechanically more brittle, while histomorphometry showed that both osteoclasts and osteoblasts had impaired activity relative to the wild type (WT). Here, we report detailed mineral and matrix analyses of the tibia of these animals based on Fourier transform infrared microspectroscopy and imaging. At 10 weeks, there was significant hypercalcification of the calcified cartilage and cortices in the KO. Carbonate content was elevated in the KO calcified cartilage as well as cortical and cancellous bone areas. These data suggest that cathepsin K does not affect mineral deposition but has a significant effect on mineralized tissue remodeling. Since growth plate abnormalities were extensive despite reported low levels of cathepsin K expression in the calcified cartilage, we used a differentiating chick limb-bud mesenchymal cell system that mimics endochondral ossification but does not contain osteoclasts, to show that cathepsin K inhibition during initial stages of mineral deposition retards the mineralization process while general inhibition of cathepsins can increase mineralization. These data suggest that the hypercalcification of the cathepsin K-deficient growth plate is due to persistence of calcified cartilage and point to a role of cathepsin K in bone tissue development as well as skeletal remodeling.
人类组织蛋白酶K缺乏会导致致密性成骨不全症,其特征为侏儒症和骨硬化。早期对10周龄雄性组织蛋白酶K缺陷(基因敲除,KO)小鼠的研究表明,它们的骨骼在机械性能上更脆,而组织形态计量学显示,相对于野生型(WT),破骨细胞和成骨细胞的活性均受损。在此,我们报告基于傅里叶变换红外显微光谱和成像技术对这些动物胫骨进行的详细矿物质和基质分析。在10周龄时,KO小鼠的钙化软骨和皮质出现明显的钙化过度。KO小鼠钙化软骨以及皮质骨和松质骨区域的碳酸盐含量升高。这些数据表明,组织蛋白酶K不影响矿物质沉积,但对矿化组织重塑有显著影响。尽管报道称钙化软骨中组织蛋白酶K表达水平较低,但生长板异常广泛,因此我们使用一种模拟软骨内成骨但不含破骨细胞的分化鸡肢芽间充质细胞系统,以表明在矿物质沉积初始阶段抑制组织蛋白酶K会延缓矿化过程,而普遍抑制组织蛋白酶则会增加矿化。这些数据表明,组织蛋白酶K缺陷生长板的钙化过度是由于钙化软骨持续存在,并表明组织蛋白酶K在骨组织发育以及骨骼重塑中起作用。