De novo synthesis of type-I collagen in bone biopsy material.
A simple and rapid method was established for the cultivation of bone cell tissue. Human bone tissue derived from orthopaedic surgery was cultivated in the presence of 14C-proline and beta-aminopropionitrile. De novo synthesized collagen was extracted from the tissue and quantified by determination of radioactivity in the purified protein. Measurements of the oxygen consumption of the tissue provided evidence that the physiological conditions for the tissue were optimal. The tissue was vital over a period of as long as 7 days, showing normal respiration and a constant rate of collagen synthesis. The observed levels of alkaline phosphatase and acid phosphatase activity clearly demonstrated that mainly osteoblasts were involved in metabolic activity. The described system is suitable for investigations of bone cell metabolism under quasiphysiological conditions.
D000135:Acid Phosphatase / Q000378:metabolism; D000367:Age Factors; D000469:Alkaline Phosphatase / Q000378:metabolism; D001842:Bone and Bones / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured / Q000378:metabolism; D003094:Collagen / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006909:Hydroxyproline / Q000378:metabolism; D008875:Middle Aged; D010006:Osteoblasts / Q000378:metabolism*; D010101:Oxygen Consumption
D003094:Collagen; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D006909:Hydroxyproline