Effect of alginate culture and mechanical stimulation on cartilaginous matrix synthesis of rat dedifferentiated chondrocytes.
To investigate whether the application of alginate culture and mechanical stimulation will improve the synthesis of cartilaginous matrix in dedifferentiated chondrocytes, rat chondrocytes underwent dedifferentiation upon serial monolayer culture up to passage 6, and then were encapsulated in 2% alginate gel and subject to static culture. After 28 days culture in static, the beads were exposed to 48 h of mechanical stimulation with continuous agitation. The sGAG content in alginate bead was measured by alcian blue staining. The expression of collagen protein was detected using immunofluorescence. After 28 days culture in alginate bead, the dedifferentiated chondrocytes remained round in shape and re-synthesized the chondrocyte-specific matrix. Compared with static culture, mechanical stimulation induced statistically increases in the production of glycosaminoglycan (p< or =0.01), as well as in the synthesis of collagen type II protein (p< or =0.05). On the contrary, no positive expression of collagen type I protein was observed at the end of culture. Our results demonstrated that both of alginate culture and mechanical stimulation help to restore chondrocyte phenotype and promotes the synthesis of cartilaginous matrix.
Wang
Yun
Y
Groupe Ingénierie Cellulaire et Tissulaire, UMR7563 CNRS-UHP-INPL LEMTA et IFR 111, Faculté de Médecine, Vandoeuvre-lès-Nancy, France. Yun.Wang@medecine.uhp-nancy.fr
de Isla
Natalia
N
Huselstein
Céline
C
Wang
Binghua
B
Netter
Patrick
P
Stoltz
Jean-François
JF
Muller
Sylvaine
S
D000464:Alginates / Q000737:chemistry*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D048429:Cell Size; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000502:physiology*; D020219:Chondrogenesis / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000502:physiology*; D008297:Male; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D010812:Physical Stimulation / Q000379:methods; D051381:Rats; D017208:Rats, Wistar; D013314:Stress, Mechanical; D023822:Tissue Engineering / Q000379:methods*
D000464:Alginates; D016326:Extracellular Matrix Proteins