Mechanical properties of cells and extracellular matrix plays important role in many biological processes like cell proliferation, differentiation, and wound healing. Mesenchymal stem cells (MSCs) are multipotent cells, capable of differentiating into many cell-types such as cartilage, osteoblast, adipocytes, and endothelial cells. In the present study we evaluated the effect of a tripeptide on hMSC proliferation, differentiation, wound healing, and cellular elasticity as an indicator mechanobiological modulation. Atomic Force Microscopy (AFM) was employed to investigate the elastic modulus of tripeptide molecules  treated hMSCs. Treated hMSCs showed Young’s modulus of 9.8± 0.09 kP which is about 2.5- fold higher than the control group (3.9±0.47 kP). Interestingly, peptide was also found to promote proliferation, adhesion, and migration of hMSCs and ameliorated wound healing. Collectively, the present study provides a quantitative description of peptide treated hMSCs mechanical behavior, which supports the process of growth and proliferation.