Many biomedical applications, titanium dioxide (TiO₂) nanoparticles are used. They have been used for drugs delivery and targeting, skin tone protection, reinforcement materials etc. It is biocompatible and non-toxic in nature. The healing of wounds are delayed mainly due to infections. Infections make the process of wound healing complex. Nanomaterials are used for enhancing wound healing mainly due to their ability to inhibit bacterial growth. Nanoparticles of TiO₂ are synthesized using a chemical method. They are annealed at different temperature to increase the particle size. XRD (X-ray diffraction), FTIR (Fourier Transform infrared spectroscopy), and TEM (Transmission electron Microscopy) are used to classify the samples. The influence of particle size and phase transformation on fibroblast cell proliferation. F-actin, cell movement are analysed. It is found that the antimicrobial activity increases with decrease in particle size. This study confirms the utilization of TiO₂ nanoparticles for wound healing applications.