Different response of articular chondrocyte subpopulations to surface motion.
OBJECTIVE
To investigate the effect of surface motion on the gene expression of proteoglycan 4 (PRG4), hyaluronan synthases (HAS1, HAS2) and on the hyaluronan (HA) and proteoglycan 4 (PRG4) release of chondrocytes from different zones of bovine articular cartilage.


METHODS
Superficial zone, deep zone, full thickness, and superficial/deep 1:1 mixed chondrocytes were seeded into 3D polyurethane scaffolds and stimulated using our bioreactor that approximates kinematics and surface motion characteristics of natural joints. One hour of surface motion superimposed on cyclic compression was applied twice a day over 3 consecutive days. Scaffolds were cut into top and bottom sections and analyzed for gene expression of PRG4, HAS1, and HAS2.


RESULTS
Depending on the cell population, the gene expression levels increased within 8 days of culture in unloaded scaffolds, with a stronger increase in the top compared to the bottom sections. Mechanical loading further enhanced the messenger RNA (mRNA) levels in all cell types, with most pronounced up-regulations observed for the PRG4 expression in deep zone and the HAS2 expression in superficial zone cells. The effect of the biochemical and biomechanical environment appeared to be additive, resulting in highest mRNA levels in the top sections of loaded constructs. Bioreactor stimulation also enhanced the HA release in all cell populations. Full thickness chondrocytes experienced the greatest effect on HAS1 mRNA expression and HA release, indicating that the interaction between cell populations may promote HA synthesis compared to subpopulations alone.


CONCLUSIONS
Reciprocating sliding can be an efficient tool for generating tissue-engineered constructs from various chondrocyte populations by providing a functional cartilage-synovial interface.
Li
Z
Z
Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, 310027, P.R. China.
Yao
S
S
Alini
M
M
Grad
S
S
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology* / Q000378:metabolism; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry*; D005786:Gene Expression Regulation; D014453:Glucuronosyltransferase / Q000096:biosynthesis* / Q000235:genetics; D000076002:Hyaluronan Synthases; D009038:Motion; D016133:Polymerase Chain Reaction; D011509:Proteoglycans / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D013314:Stress, Mechanical; D023822:Tissue Engineering / Q000379:methods; D000099158:Proteoglycan 4
D003470:Culture Media; D014453:Glucuronosyltransferase; D000076002:Hyaluronan Synthases; D011509:Proteoglycans; D012333:RNA, Messenger; C419197:PRG4 protein, human; D000099158:Proteoglycan 4