VEGF production by osteoarthritic chondrocytes cultured in micromass and stimulated by IL-17 and TNF-alpha.
We compared the secretion of vascular endothelial growth factor (VEGF) by chondrocytes isolated from cartilage of patients with osteoarthritis (OA) and maintained in monolayer or in three-dimensional culture. Chondrocytes, immediately after isolation, or after one passage in monolayer culture (subculture), were seeded in monolayer or pelleted in micromasses. Cells cultured differently were immunoassayed for the secretion of VEGF in basal conditions or after stimulation with IL-17, TNF-alpha or B IL-17+TNF-alpha. Chondrocytes cultured in micromasses secreted lower levels of VEGF in comparison with those in monolayer culture. In micromass cultures TNF-alpha was more stimulating than IL-17 and their combined effect was additional. On the basis of alkaline phosphatase/cathespin B activities, primary cultures in micromass, characterized by low VEGF secretion, represented the best condition for maximal cell differentiation. Our results suggest that the culture of chondrocytes in micromasses represents a good condition to study the physiology of these cells to a maximum degree when micromasses are obtained with freshly isolated chondrocytes.
Honorati
Maria Cristina
MC
Dipartimento di Medicina Interna e Gastroenterologia, Università di Bologna, Bologna, Italy. labimge@alma.unibo.it
Cattini
Luca
L
Facchini
Andrea
A
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000469:Alkaline Phosphatase / Q000378:metabolism; D002401:Cathepsin B / Q000378:metabolism; D018929:Cell Culture Techniques; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D020381:Interleukin-17 / Q000494:pharmacology*; D008875:Middle Aged; D010003:Osteoarthritis / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis*
D020381:Interleukin-17; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D000469:Alkaline Phosphatase; D002401:Cathepsin B