Articular chondrocytes from osteoarthritic knee joints of elderly, in vitro expanded in thermo-reversible gelation polymer (TGP), exhibiting higher UEA-1 expression in lectin microarray

Shojiro Katoh; Atsuki Fujimaru; Rajappa Senthilkumar; Senthilkumar Preethy; Samuel Jk Abraham

doi:10.1016/j.reth.2020.03.006

Articular chondrocytes from osteoarthritic knee joints of elderly, in vitro expanded in thermo-reversible gelation polymer (TGP), exhibiting higher UEA-1 expression in lectin microarray

Regen Ther. 2020 May 15:14:234-237. doi: 10.1016/j.reth.2020.03.006. eCollection 2020 Jun.

Authors

Shojiro Katoh^{1

2}, Atsuki Fujimaru², Rajappa Senthilkumar³, Senthilkumar Preethy³, Samuel Jk Abraham^{4

5

6

7}

Affiliations

¹ Edogawa Evolutionary Lab of Science, Edogawa Hospital Campus, 2-24-18, Higashikoiwa, Edogawa, Tokyo 133-0052, Japan.
² Department of Orthopaedic Surgery, Edogawa Hospital, 2-24-18, Higashikoiwa, Edogawa, Tokyo 133-0052, 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.
⁵ JBM Inc., 3-1-14, Higashikoiwa, Edogawa, Tokyo 133-0052, Japan.
⁶ Yamanashi University-Faculty of Medicine, 1110, Shimokato, Chuo, Yamanashi 409-3898, Japan.
⁷ GN Corporation Co. Ltd., 3-8, Wakamatsu, Kofu, Yamanashi 400-0866, Japan.

Abstract

Autologous chondrocytes in vitro expanded, are used as tools of regenerative therapies for cartilage injuries. However, inability to maintain the hyaline phenotype both in vitro and post in vivo transplantation, remains one of the major hurdles for long term efficacy under clinical settings. We have reported earlier, hyaline phenotype maintenance of both human and rabbit chondrocytes for a long duration both in vitro when cultured conditions using a Thermo-reversible Gelation Polymer (TGP) scaffold-based methodology and in vivo post-transplantation animal model of cartilage damage. Having intrigued by such encouraging outcome, we in this study, analysed the similar TGP culture environment whether would be able to allow in vitro expansion of severe osteoarthritis affected cartilage tissue from elderly patients and evaluated the cells using lectin microarray characterization for pluripotency. Cartilage tissue were obtained from patients (n = 7; age: 60-85 years) undergoing total knee arthroplasty for severe osteoarthritis. Chondrocytes were isolated and cultured in two groups: i. conventional culture without scaffold (2D) and ii. using a TGP scaffold-based culture (3D) up to 18 weeks. In addition to earlier reported findings such as maintenance of hyaline phenotype having been confirmed in this study as well, surface glycoprotein analysis by lectin microarray demonstrated that the α1-2 Fuc recognition lectin (UEA-1) (marker reported in literature for pluripotent stem cells) was found to be more highly expressed in 3D culture compared to 2D culture and even increased over time in 3D culture. We have developed an environment where osteoarthritis affected chondrocytes from the elderly could be cultured up to 18 weeks in vitro using TGP scaffold which express pluripotent cell associated surface glycoproteins compared to the conventional methodology.

Keywords: 2D, Two-dimensional; 3D, Three-dimensional; ACI, Autologous chondrocyte implantation; CO2, Carbon dioxide; Cartilage; Chondrocytes; ESC, Embryonic stem cells; Hyaline phenotype; Lectin microarray; MACI, matrix-associated chondrocyte implantation; Osteoarthritis; PBS, Phosphate-buffered saline; Pluripotency; TGP, Thermo-reversible gelation polymer; Thermo-reversible gelation polymer (TGP) scaffold; hPSCs, Human pluripotent stem cells; iPSC, Induced pluripotent stem cells.