Abstract
BackgroundIn order to optimize the tenogenic differentiation of mesenchymal stem cells (MSCs), researchers should consider various factors. However, this requires testing numerous experimental settings, which is costly and time-consuming. We aimed to assess the differential effects of transforming growth factor beta-3 (TGF-β3) on the tenogenesis of tonsil-derived MSCs (T-MSCs) and bone marrow-derived MSCs (BM-MSCs) using response surface methodology (RSM).MethodsBone marrow and tonsillar tissue were collected from four patients; mononuclear cells were separated and treated with 5 or 10 ng/mL of TGF-β3. A full factorial experimental design with a categorical factor of 0 was employed to study the effect of tension based on T-MSCs. Eighty-four trials were fitted with RSM and then used to obtain mathematical prediction models.ResultsExposure of T-MSCs and BM-MSCs to TGF-β3 increased the expression of scleraxis (SCX), tenomodulin (TNMD), decorin, collagen I, and tenascin C. Expression of most of these factors reached a maximum after 2–3 days of treatment. The model predicted that the values of the tenocyte lineage-related factors assessed would be significantly increased at 2.5 days of culture with 2.7 ng/mL of TGF-β3 for T-MSCs and at 2.3 days of culture regardless of TGF-β3 concentration for BM-MSCs.ConclusionsThis study demonstrated that the RSM prediction of the culture time necessary for the tenogenic differentiation of T-MSCs and BM-MSCs under TGF-β3 stimulation was similar to the experimentally determined time of peak expression of tenocyte-related mRNAs, suggesting the potential of using the RSM approach for optimization of the culture protocol for tenogenesis of MSCs.
Highlights
In order to optimize the tenogenic differentiation of mesenchymal stem cells (MSCs), researchers should consider various factors
Bone marrow-derived, adipose tissue-derived, synovial membrane-derived, and human embryonic MSCs have all been used to differentiate into tenocytes [6, 9, 10], these stem cells are obtained through invasive procedures
Tonsil-derived MSCs (TMSCs) obtained from waste tissue after tonsillectomy represent a new source of progenitor cells [12, 13], and several studies have focused on tonsil-derived MSCs (T-MSCs) as cellular therapeutic agents for various diseases [12,13,14,15,16,17,18,19]
Summary
In order to optimize the tenogenic differentiation of mesenchymal stem cells (MSCs), researchers should consider various factors. The structure and strength of the repaired tissue do not show full recovery, and the tissue does not return to the pre-injury state [4] For these reasons, there have been various studies done on tendon repair using tissue engineering with mesenchymal stem cells (MSCs) [5,6,7,8,9]. A previous study reported that T-MSCs have a good potential of being suitable for clinical banking of stem cells [12] This means that it might be possible to conduct tonsil tissue banking (after tonsillectomy) for autogenic MSCs grafts to be used in case of future injury or disease. Protocols for tenogenic differentiation of T-MSCs have not been established, and there is a lack of studies comparing the tenogenic differentiation potential of TMSCs with MSCs from other cell sources
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