A high expression of hyaluronic acid (HA) in precartilage condensation at the early stage of endochondral ossification indicates its importance in bone and cartilage development. However, it remains unclear whether HA is associated with bone development by promoting the differentiation from mesenchymal stem cells (MSCs). In this study, we examined the effects of HA on the differentiation of MSCs into osteoblasts and chondrocytes in a three-dimensional collagen gel culture system.

Human MSCs were cultured in HA-collagen hybrid gel (0.15% collagen, 0.5 mg/ml HA) and collagen mono gel (0.15% collagen). The cells were treated by either osteogenic differentiation medium (ODM) or chondrogenic differentiation medium (CDM). The

* Address correspondence and reprint requests to: Eiji Tanaka, Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, 12-3 Kasumi, Minami-Ku, Hiroshima 734-8553, Japan. Tel: +81-82-257-5686. Fax: +81-82-257-5687. Email: [email protected]

HA-collagen gel maintained in ODM exhibited greater alkaline phosphatase activity, calcium deposition, and gene expression of bone markers than the collagen mono gel. The HA-collagen gel maintained in CDM had greater glycosaminoglycan deposition and gene expression of cartilage markers than the collagen mono gel.

These findings indicate that the addition of HA to collagen gel would be highly recommended for promoting the differentiation of MSC to osteoblasts and chondrocytes in three-dimensional condition.

Keywords: human mesenchymal stem cell; three-dimensional collagen gel; osteogenesis; chondrogenesis; hyaluronic acid

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