Markedly different effects of hyaluronic acid and chondroitin sulfate‐A on the differentiation of human articular chondrocytes in micromass and 3‐D honeycomb rotation cultures

Abstract

A source of morphologically and functionally available human cartilagenous tissue for implantation is required in the field of tissue engineering. To achieve this goal, we evaluated the effects of hyaluronic acid (HA-810 and 1680 kDa), and chondroitin sulfate (CS-A 16 and C-34 kDa) on human articular chondrocytes (HC) in micromass and rotation culture conditions. Cell proliferation was increased by CS-A 16 kDa under micromass and rotation cultures, while cell differentiation was increased under rotation but not micromass conditions. Proliferation and differentiation due to CS-C 34 kDa were very similar to the control under both culture conditions. With HA, cell proliferation was increased depending on the molecular weight under micromass and rotation conditions. In contrast, chondrocyte differentiation was enhanced under rotation conditions, but decreased under micromass conditions depending on the molecular weight of HA. In both culture conditions, aggrecan gene was continuously expressed. However, the collagen type II gene was more weakly expressed in rotation than the micromass culture conditions. Thus, the chemical structures of polysaccharides, and the culture condition, rotation or micromass, caused differences in chondrogenesis.

Pellet culture elicits superior chondrogenic redifferentiation than alginate‐based systems

“…1,[4][5][6][7][8][9][10][11][12][13][14][15]17,27,28,30,31 Collagen I production was generally broad-ranged in all biomaterial encapsulated chondrocytes, which might be due to the ubiquitous stimulative properties of TGF-b that was used as an ingredient for the cell culture medium used in our study. 32 Besides certain outliers, a trend towards higher Collagen I expression in alginate-fibrin encapsulated chondrocytes could be observed, whereas Collagen I expression was constantly low in pellet-cultured cells.…”

“…It is known that biomaterial-free pellet culture can elicit profound chondrogenic effects upon dedifferentiated chondrocytes or mesenchymal stem cells. [13][14][15]17,18,[27][28][29] A major drawback of those techniques is the large amount of cells that is needed to generate sufficiently sized cartilage constructs.…”

“…7 Coming from the observation of mesenchymal condensation in the developing limb bud, pellet culture systems had been established to differentiate mesenchymal stem cells 13,14 and dedifferentiated chondrocytes 15,16 into hyaline cartilage pellets. Although the micro mass system is performing excellently in terms of chondrogenic differentiation, its application for clinical use in tissue engineering is in doubt, mainly due to the limited size and instability of the constructs in vivo.…”

Addition of hyaluronic acid improves cellular infiltration and promotes early-stage chondrogenesis in a collagen-based scaffold for cartilage tissue engineering

“…Studies previously carried out 5 on cartilage regeneration have shown that GAGs are fundamental for presenting motifs for cell attachment and migration. In addition, GAGs have also been attributed to activating pathways that govern proliferation and differentiation as well as assembly of the ECM (Pieper et al, 2000, Banu and Tsuchiya, 2007, Wu et al, 2010, Correia et al, 2011. Specifically, chondroitin sulphate (CS) which is the main type of GAG in cartilage ECM has been shown to stimulate chondrogenesis in vitro and promotes cellular ingrowth and cartilaginous tissue formation in vivo (van Susante et al, 2001, Buma et al, 2003.…”

Using chondroitin sulfate to improve the viability and biosynthesis of chondrocytes encapsulated in interpenetrating network (IPN) hydrogels of agarose and poly(ethylene glycol) diacrylate

“…There have been a number of papers published on the subject of crosslinking GAGs for biomedical use [26–34]. A good example of this approach is given by the work of Li et al [29].…”