Shojiro Katoh1, Hiroshi Yoshioka2, Rajappa Senthilkumar3, Senthilkumar Preethy3, Samuel J K Abraham4. 1. Edogawa Evolutionary Lab of Science, Edogawa Hospital Campus, 2-24-18, Higashi Koiwa, Edogawa, Tokyo 133-0052, Japan; Department of Orthopaedic Surgery, Edogawa Hospital, 2-24-18, Higashi Koiwa, Edogawa, Tokyo 133-0052, Japan. 2. Mebiol Inc., 1-25-8, Nakahara, Hiratsuka 254-0075, Kanagawa, Japan. 3. The Fujio-Eiji Academic Terrain (FEAT), Nichi-In Centre for Regenerative Medicine (NCRM), PB 1262, Chennai 600034, Tamil Nadu, India. 4. The Mary-Yoshio Translational Hexagon (MYTH), Nichi-In Centre for Regenerative Medicine (NCRM), PB 1262, Chennai 600034, Tamil Nadu, India; JBM Inc., 3-1-14, Higashi Koiwa, Edogawa, Tokyo 133-0052, Japan; University of Yamanashi -Faculty of Medicine, 1110, Shimokato, Chuo, Yamanashi 409-3898, Japan; GN Corporation Co. Ltd., 3-8, Wakamatsu, Kofu, Yamanashi 400-0866, Japan. Electronic address: drsam@nichimail.jp.
Abstract
AIMS: We have evaluated the potential of a three-dimensional (3D) thermoreversible gelation polymer (TGP) matrix in enhancing miRNA 140 expression (a biomarker correlating with homeostasis and cartilage regeneration) during the in vitro expansion of osteoarthritis (OA)-affected human chondrocytes. MATERIALS AND METHODS: OA-chondrocytes were cultured in two-dimensional (2D) monolayer followed by culture in 3D-TGP. miRNA 140 expression levels in cell culture supernatant followed by expression in the cell lysate of both 2D and 3D-TGP cultures were analyzed. KEY FINDINGS: The expression of miRNA 140 in cell culture supernatant from the 3D-TGP group was 0.001 to 0.002% that in 2D culture supernatant while in the cell lysate, miRNA 140 expression in the 3D-TGP was nearly 30-fold higher than that of 2D group. SIGNIFICANCE: The 3D-TGP matrix allows enhanced expression of miRNA 140 in OA-affected human chondrocytes in vitro which after necessary validations can be applied in clinical transplantation to significantly improve the outcome.
AIMS: We have evaluated the potential of a three-dimensional (3D) thermoreversible gelation polymer (TGP) matrix in enhancing miRNA 140 expression (a biomarker correlating with homeostasis and cartilage regeneration) during the in vitro expansion of osteoarthritis (OA)-affected human chondrocytes. MATERIALS AND METHODS: OA-chondrocytes were cultured in two-dimensional (2D) monolayer followed by culture in 3D-TGP. miRNA 140 expression levels in cell culture supernatant followed by expression in the cell lysate of both 2D and 3D-TGP cultures were analyzed. KEY FINDINGS: The expression of miRNA 140 in cell culture supernatant from the 3D-TGP group was 0.001 to 0.002% that in 2D culture supernatant while in the cell lysate, miRNA 140 expression in the 3D-TGP was nearly 30-fold higher than that of 2D group. SIGNIFICANCE: The 3D-TGP matrix allows enhanced expression of miRNA 140 in OA-affected human chondrocytes in vitro which after necessary validations can be applied in clinical transplantation to significantly improve the outcome.