The optimal mechanical condition in stem cell-to-tenocyte differentiation determined with the homogeneous strain distributions and the cellular orientation control

In tendon tissue engineering, mechanical stimulus-induced differentiation is one of the most attractive techniques for stem cell-to-tenocyte differentiation in terms of cost, safety and simplicity. However, the most effective strain amplitude for differentiation using cyclic stretching remains unknown. Existing studies have not constrained cell reorientation behavior during cyclic stretching, resulting in uncertainty regarding the loads experienced by cells. In addition, strain distribution homogeneity of the culture membrane is important. Here, we improved the strain distribution uniformity of the membrane and employed a microgrooved membrane to suppress cell reorientation. Then we evaluated the most effective strain amplitude (0, 2, 4, 5, 6, or 8%) for the differentiation of mesenchymal stem cells into tenocytes by measuring mRNA expression levels. The maximum expression of all tenogenic markers was observed at a 5% strain. These results contribute to tendon tissue engineering by clarifying the most effective strain amplitude during tenogenic differentiation induction using cyclic stretching.