Date of Award

11-11-2024

Thesis Type

Masters

Document Type

Dissertation

Divisions

Faculty of Medicine

Department

Orthopaedic Surgery

Institution

Universiti Malaya

Abstract

Tendon injuries represent a significant challenge to treat owing to their limited intrinsic reparative capacity. The use of mesenchymal stem cells (MSC) offers promising alternative therapeutic option to augment tendon repair. It is hypothesised that the activation of hypoxia inducible factor-1 alpha (HIF-1α), could facilitate the tendon repair process by promoting the proliferation and tenogenic differentiation of MSCs. To demonstrate this, a study was conducted incorporating the use of Roxadustat, a specific hypoxia mimetic mediator and cyclic uniaxial stretching at a frequency of 1 Hz and 8 % strain on adipose derived-mesenchymal stromal cells (ADMSCs). Methods: Cellular morphology, proliferation rate, tenogenic protein and gene expression levels from 8 different treatment groups were compared. These groups include (1) untreated ADMSCs (Control), (2) Roxadustat pre-conditioned ADMSCs (ROX), (3) ADMSCs subjected to CAY10585 treatment only (CAY), (4) Roxadustat pre-conditioned ADMSCs with CAY10585 inhibition (ROX+CAY), (5) ADMSCs subjected to cyclic uniaxial stretching only (S), (6) Roxadustat pre-conditioned ADMSCs with cyclic uniaxial stretching (ROX+S), (7) ADMSCs subjected CAY10585 with cyclic uniaxial stretching (CAY+S) and (8) primary tenocytes (Tenocytes). Results: MSCs pre-conditioned with 12.5μM Roxadustat for 24-hour showed a significant increase in expression of HIF-1α without affecting the proliferation rate of ADMSCs. A significant reduction of HIF-1α levels at 12.5μM concentration was observed when the cells were treated with 3.5μM CAY10585, as seen between ROX and ROX+CAY group. In addition, ROX+S group exhibited the highest expression of HIF-1α and demonstrated a significant up regulation of collagen I and III expressions, increasing by 4.9 and 5.6-fold compared to ROX group, respectively. There is a significant increase of SCX, TNC, TNMD, COLI and COLIII expression in this combination treatment group; (SCX= 9.9, TNC= 12.6, TNMD= 7.0, COLI= 8.0 and COLIII= 10.0-fold). Conversely, the expression of the markers markedly reduced with HIF-1α inhibitor CAY10585. However, cyclic uniaxial stretching effectively counteracted the inhibitory effects of CAY10585 in the CAY+ S group, resulting in a 3.9-fold increase in SCX expression compared to CAY treatment alone. Conclusion: HIF-1α accumulation promotes superior tenogenic differentiation of ADMSCs, suggesting that the combination of Roxadustat and cyclic uniaxial stretching may be a potential therapeutic mediator in tendon repair strategies.

Initial

khm 

Additional Information

Dissertation (M.A.) – Faculty of Medicine, Universiti Malaya, 2024.

Download

Available for download on Thursday, December 31, 2026

Share

COinS