title: Nonenzymatic glycation of type I collagen modifies interaction with UMR 201-10B preosteoblastic cells.
abstract: Advanced glycation endproduct (AGE), whose formation is accelerated on long lived extracellular matrix proteins in diabetes, is implicated in diabetic complications in various tissues. Type I collagen is the predominant matrix protein of bone and plays an important role in bone cell-matrix interactions. We have previously reported the accelerated accumulation of AGE collagen in bone tissue in diabetes mellitus (DM), in which reduced bone mineral density was observed. In addition, when cultures of mature primary rat osteoblasts were plated onto an in vitro AGE-modified collagen substrate, they showed altered cell functions, in terms of alkaline phosphatase (ALP) activity, osteocalcin secretion, and nodule formation (J Bone Miner Res 11:931-937; 1996). To determine whether AGE collagen might also affect differentiation of preosteoblasts, we compared the effects of plating the preosteoblastic UMR 201-10B cell line onto AGE-modified collagen with plating onto unmodified collagen. The latter had been shown previously to promote differentiation of UMR 201 cells. We have also explored whether these effects might be partly mediated by the transforming growth factor beta (TGF-beta) receptor. Growth of UMR 201-10B cells on a type I collagen substrate significantly inhibited cell growth and retinoic acid (RA)-induced upregulation of ALP activity, compared to cells on plastic. These inhibitory effects were reduced by prior glycation of collagen, in a dose-dependent manner with respect to AGE content. Unmodified collagen stimulated production of osteopontin mRNA, which was reduced by AGE modification to levels attained in cells on plastic. Growth on control collagen inhibited TGF-beta type II receptor binding in 10B cells, while this inhibition was reduced by AGE modification. These data suggest that glycation of collagen interferes with specific interaction(s) between UMR 201-10B cells and collagen. Based on our previous results in UMR 201 cells, these results would be compatible with the notion that glycated collagen has reduced ability to promote differentiation of preosteoblasts to mature osteoblasts. These data further suggest that collagen-mediated events in these cells may be at least in part mediated by regulation of the TGF-beta receptor expression.
authors.affiliation: St. Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.
authors.forename: Y
authors.initials: Y
authors.lastname: Katayama
authors.forename: S
authors.initials: S
authors.lastname: Celic
authors.forename: N
authors.initials: N
authors.lastname: Nagata
authors.forename: T J
authors.initials: TJ
authors.lastname: Martin
authors.forename: D M
authors.initials: DM
authors.lastname: Findlay
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003094:Collagen / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D003432:Cross-Linking Reagents; D005786:Gene Expression Regulation / Q000187:drug effects; D019298:Glucose-6-Phosphate / Q000494:pharmacology; D017127:Glycation End Products, Advanced / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D053495:Osteopontin; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism*; D012795:Sialoglycoproteins / Q000378:metabolism; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D003432:Cross-Linking Reagents; D017127:Glycation End Products, Advanced; D012333:RNA, Messenger; D018125:Receptors, Transforming Growth Factor beta; D012795:Sialoglycoproteins; C506897:Spp1 protein, rat; D016212:Transforming Growth Factor beta; D053495:Osteopontin; D019298:Glucose-6-Phosphate; D014212:Tretinoin; D003094:Collagen; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II; D000469:Alkaline Phosphatase