Synthetic peptide containing Arg-Gly-Asp inhibits bone formation and resorption in a mineralizing organ culture system of fetal rat parietal bones.
The role of integrins, cell surface receptors involved in cell adhesion to the matrix, was studied in a mineralizing organ culture system. Integrin-mediated cell attachment to matrix proteins has been shown to depend partially on the amino acid sequence Arg-Gly-Asp (RGD), present in the extracellular matrix proteins. Therefore, the effect of RGD peptides on bone formation and resorption was studied in the mineralizing organ culture system derived from 18 day fetal rat parietal bones. Addition of 0.1-50 microM GRGDSPK to bones cultured for 4 days inhibited mineralization in a dose-dependent manner as determined by measuring calcium content and % bone/unit area of tissue. A maximal decrease in calcium content and % bone/unit area of 32.5 and 42.9%, respectively, was found with 50 microM GRGDSPK. With 10 and 50 microM GRGDSPK, bone morphology was dramatically altered, with a disruption of osteoblast and mineralized matrix organization. To assess the effect of the peptides on bone resorption, fetal bones were prelabeled in vivo with 45Ca and resorption was stimulated in vitro with parathyroid hormone in the presence or absence of the peptide. A significant decrease in 45Ca release was found with 10 and 50 microM GRGDSPK. Osteoclast number was also significantly decreased on the bone surface. The peptide was not cytotoxic, since no effect on DNA content, dry weight, or collagen synthesis was found. GRADSP, a control peptide, had no significant effect on mineralization, resorption, or other parameters of bone growth.(ABSTRACT TRUNCATED AT 250 WORDS)
Gronowicz
G A
GA
Department of Orthopaedics, University of Connecticut Health Center, Farmington.
Derome
M E
ME
D000595:Amino Acid Sequence; D000818:Animals; D001862:Bone Resorption / Q000503:physiopathology*; D002113:Calcification, Physiologic / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D003094:Collagen / Q000096:biosynthesis; D005260:Female; D016023:Integrins / Q000032:analysis; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology*; D009924:Organ Culture Techniques; D010006:Osteoblasts / Q000187:drug effects / Q000502:physiology; D010010:Osteoclasts / Q000187:drug effects / Q000502:physiology; D010012:Osteogenesis / Q000187:drug effects*; D010281:Parathyroid Hormone / Q000494:pharmacology; D010294:Parietal Bone / Q000196:embryology; D011247:Pregnancy; D051381:Rats
D016023:Integrins; D009842:Oligopeptides; D010281:Parathyroid Hormone; C086456:glycyl- arginyl-glycyl-aspartyl-seryl-prolyl-lysine; C047981:arginyl-glycyl-aspartic acid; D003094:Collagen; D002118:Calcium