2021 ISAKOS Biennial Congress ePoster
Enhanced miRNA 140 and Hyaluronic acid expression in in vitro cultured human chondrocytes derived from OA affected cartilage tissue; An ideal tool for ACI and MACI
Samuel Jk Abraham, MD, PhD, Kofu, Yamanashi JAPAN
Hiroshi Yoshioka, MD, Tokyo JAPAN
Senthilkumar Rajappa, PhD, Chennai INDIA
Senthilkumar Preethy, BDS, MSc.,, Chennai INDIA
Karthick Ramalingam, MSc, Chennai INDIA
Rajmohan Mathaiyan, MSc, Chennai INDIA
Akimasa Seno, PhD, Okayama JAPAN
Masaharu Seno, PhD, Okayama JAPAN
Shojiro Katoh, MD, Tokyo JAPAN
Yamanashi University, Chuo, Yamanashi, JAPAN
The FDA has not cleared the following pharmaceuticals and/or medical device for the use described in this presentation. The following pharmaceuticals and/or medical device are being discussed for an off-label use: JBM Inc, GN Corporation Inc, Chondrocyte and polymer mix; Mebiol Inc, Thermo-reversible gelation polymer
Summary
In vitro cultured chondrocytes with enriched hyaluronic acid and miRNA140 as ideal tools for cell therapy applications such as ACI and MACI
ePosters will be available shortly before Congress
Abstract
Background
In vitro expanded chondrocytes being tools of cell therapy in ACI and MACI have been evaluated for their phenotype and gene expression. In this study we report a 3D polymer scaffold platform which yields chondrocytes with enriched hyaluronic acid (HA) content and miRNA 140 which are considered hallmarks of healthy chondrocytes and are essential for maintenance of the homeostasis of cartilage tissue.
Materials And Methods
Osteo-arthritis (OA) affected knee cartilage, removed before TKA were used in this study (n=7). Small bits of the tissue, observed to be healthy by the orthopedician were harvested, (weighing 0.1 ~0.3 grams) enzyme digested and the isolated chondrocytes were cultured in monolayer (2D) and in a Thermoreversible gelation polymer (3D-TGP). 2D specimens were cultured until they degenerated and 3D group till 42nd day. No growth factors or fetal calf serum or eternally added hyaluronic acid were used during the entire culture duration. Cells were harvested and subjected to evaluation of HA and miRNA 14-0 expression by RT PCR and Immunohistochemistry.
Results
In 2D, the cells grew with fibroblast morphology whereas in 3D they started forming clusters and grew as tissues. miRNA 140 expression in 3D was 30-fold higher than 2D and the HA expression evaluated by staining for CD44 antibody was more intense (IHC score = +++) compared to the 2D (IHC score = +).
Conclusion
To our knowledge this is the first of its kind report in the literature reporting HA and miRNA enrichment in cultured human chondrocytes taken from damaged and inflamed OA affected cartilage, thus making the 3D platform a suitable for producing ideal type of cells for ACI and MACI. Further, encapsulation of cells with the scaffold during transplantation is worth further research as it may yield a better in vivo environment for enhanced engraftment and hence better clinical results.