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The development of bone canaliculi in the endochondral or intramembranous ossification analyzed by histological study of decalcified bone using silver staining method



The development of bone canaliculi in the endochondral or intramembranous ossification analyzed by histological study of decalcified bone using silver staining method



Journal of Kyoto Prefectural University of Medicine 100(8): 801-818



The silver impregnation staining with protargol (silver protein), which is a modified Bodian method, has been found to be useful to identify histologically the details of bone canaliculi structure in the decalcified bone tissue. Using this staining method, the author studied the development of bone canaliculi structure built up by osteocytes during endochondral or intramembranous ossification. Endochondral ossification was observed at metaphysis beneath the growth plate of proximal tibia of growing rat, and the process of intramembranous ossification was detected in the callus tissue of the diaphysis of rat fermur obtained from experimental fracture model (drill-hole model). The results of this study revealed that woven bone osteocytes formed not only fewer but also much more irregular bone canaliculi in the matrix than lamellar bone osteocytes. It is therefore apparent that the morphological distinction between woven bone osteocytes and lamellar bone osteocytes is much easier and more accurate in the decalcified bone section stained with protargol than in that stained with hematoxylin eosin. It was also demonstrated that the woven bone osteoblasts were able to attach to both the surface of degenerative cartilage matrix in the calcified zone of growth plate and also to the surface of the injured cortical bone matrix after drilling of bone. In both the endochondral and intramembranous ossification, surface of the thin woven bone built up by woven bone osteocytes was lined with many lamellar bone osteoblasts which would differentiate into lamellar bone osteocytes with numerous bone canaliculi running regularly in the calcified bone matrix. The woven bone seemed to play a role of scaffold for lamellar bone formation. Such newly found lamellar bone became thick by appositional growth of lamellar bone osteocytes. This suggests that woven bone is able to induce a larger amount of lamellar bone, or that active lamellar bone formation needs woven bone at the early stages of ossification. It is thus likely that woven bone acts as a mediator between degenerative cartilage or injured bone and lamellar bone. It is concluded that woven bone is, morphologically and biologically, different tissue from lamellar bone, and also that the present method of silver staining is useful to distinguish them.

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Accession: 007882077

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