1990 Fiscal Year Final Research Report Summary
Celluar Biological Analysis on the Mechanism of Mineralization of Human Bone Tissue and its Application for Reconstruction of Human Tissue in Vitro
| Project/Area Number |
62440077
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
外科・放射線系歯学
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| Research Institution | Tokushima University |
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Principal Investigator |
NAGAYAMA Masaru Tokushima Univ., School of Dentistry, Professor, 歯学部, 教授 (30022867)
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| Co-Investigator(Kenkyū-buntansha) |
GOTOH Yuji Tokushima Univ., School of Dentistry, Research Assoc., 歯学部, 助手 (60225670)
TAKISHITA Yukio Tokushima Univ., School of Dentistry, Research Assoc., 歯学部, 助手 (30226964)
HAYASHI Eiji Tokushima Univ., Univ. Dental Hospital, Research Assoc., 歯学部・附属病院, 助手 (50173000)
YASUDA Katsuhiro Tokushima Univ., School of Dentistry, Research Assoc., 歯学部, 助手 (00174508)
HIRAIWA Kiyotaka Tokushima Univ., Univ. Dental Hospital, Assist. prof., 歯学部・附属病院, 講師 (30173214)
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| Project Period (FY) |
1987 – 1990
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| Keywords | human osteoblastic cells / collagen gel / platelet-derived growth factor / bone marrow stromal cell / in vitro mineralization / chondrocytes / retinoic acid / endochondral ossification |
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| Research Abstract |
Osteoblastic cells were isolated from human maxilla by embedding the bone pieces in collagen gel. As a result of the culture and characterization of the isolated cells, the cells responded to PTH and PGE_2, secreted osteocalcin and formed mineralized area in monolayer culture. When we investigated the effect of several factors on differentiation and proliferation of the cells, platelet-derived growth factor contained in the bone matrix tended to stimulate proliferation of the cells.
Bone marrow tissue is reported to contain cells which can differentiate to osteoblast and chondroblast. From these facts, we cultured bone marrow stromal cells and elucidated that the tissue has osteogenic cells, which formed mineralized nodules in vitro. The nodules were analyzed histologically and immunohistochemically. Among the cells, it is confirmed to exist the cells which could mineralize without beta-glycerophosphate.
As chondrocytes have an important role in bone formation, we have also studied the f
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ollowing research. We obtained terminally differentiated chondrocytes in monolayer culture. The cells tended to be stimulated proliferation by retinoic acid. The results suggested that the mechanism of proliferation by retinoic acid was not mediated by epidermal growth factor but by protein kinase C. The retinoic acid binding capacity was detected in both the cytosol and nuclear of the cultured chondrocytes. These binding capacity was supposed to be involved in the mechanism of retinoic acid.
Furthermore, in order to obtain more insight into the physiologic mechanism of endochondral ossification, changes occurring in the mandibular cartilage of growing rats fed on a low-calcium diet were investigated. The results indicated that calcium-deficiency and subsequent metabolic disorder in growing rats induced inhibition of cell proliferation in the proliferative cell zone, inhibition of maturation of chondrocytes, and inhibition of cartilage resorption due to poor calcification of the extracellular matrix, so that the mechanism of normal endocondral ossification might be inhibited and enibited and endochondral growth of mandibular condyle by suppressed. Less
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