Production of collagenase and tissue inhibitor of metalloproteinases (TIMP) by rat growth plates in culture.
Growth plate cartilage from normal and vitamin D-phosphate deficient (-VDP) rats was cultured to study the production of collagenase and tissue inhibitor of metalloproteinases (TIMP) in vitro. All tissues secreted latent collagenase into the medium at a constant rate during the 5 days in culture. Microdissected-VDP growth plates, containing predominatly hypertrophic cells, released up to 8-fold more collagenase into the medium than either intact-VDP or normal growth plates. TIMP was also secreted during the culture, but its rate of production was not as dependent on tissue type as collagenase. The tissue level of collagenase and TIMP before culture was compared with that found in conditioned medium and remnant tissue after culture. During the 5 day culture period microdissected-VDP growth plates, containing predominatly hypertrophic cells, produced 3-times more collagenase/microgram DNA over the starting level than either intact-VDP or normal growth plates. TIMP was never found in tissues after they had been cultured, but was present in all tissues before culture except those containing predominatly hypertrophic cells. The amount of TIMP required to block collagenase was calculated. Growth plates in culture produced enough TIMP to block all collagenase found in the medium and remnant tissue, while extracts of uncultured intact -VDP growth plates, and those divided to contain hypertrophic cells, had an excess of collagenase over TIMP. The results suggest that hypertrophic cells produce far more collagenase than other cells in the growth plate, but all cell types have about the same capacity to synthesize TIMP. As a result, increased collagenase synthesis by hypertrophic cells may surpass increases in TIMP synthesis and lead to collagen removal. This would allow for thinning of the longitudinal septa and expansion of the hypertrophic cells.
Dean
D D
DD
Arthritis Research Laboratory, U.S. Veterans Administration Medical Center.
Muniz
O E
OE
Woessner
J F
JF
Howell
D S
DS
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002850:Chromatography, Gel; D003470:Culture Media / Q000494:pharmacology; D004247:DNA / Q000032:analysis; D006023:Glycoproteins / Q000096:biosynthesis*; D006132:Growth Plate / Q000378:metabolism*; D006909:Hydroxyproline / Q000032:analysis; D008297:Male; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D009924:Organ Culture Techniques; D010710:Phosphates / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D019714:Tissue Inhibitor of Metalloproteinases
D003470:Culture Media; D006023:Glycoproteins; D010710:Phosphates; D019714:Tissue Inhibitor of Metalloproteinases; D004247:DNA; D008666:Metalloendopeptidases; D003012:Microbial Collagenase; D006909:Hydroxyproline; D002118:Calcium