This work presents a design of experiment (DoE) approach to optimize the electrophoretic deposition (EPD) parameters for chitosan (CS), gelatin (Gel), and whitlockite (WH) composite coatings on titanium substrate. The suspension for EPD was prepared by dissolving CS and Gel in acetic acid and WH nanoparticles (NPs) were dispersed in ethanol. Afterward, both (CS, Gel solution, and WH suspension) were mixed and the suspension stability was evaluated using zeta potential. Deposition parameters like voltage, concentration of WH, and time were optimized using Taguchi L16 orthogonal array with four control factors. The statistical data obtained confirmed that a higher time with average voltage and WH concentration is ideal to achieve deposits with maximum yield and minimum signal-to-noise ratio. Tea tree oil (TTO) in different concentrations was added to these optimized conditions. Different physiochemical characterization like x-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), in-vitro degradation, surface adhesion, and wettability tests were done to evaluate the coating stability, adhesion, surface and chemical properties. The obtained data confirmed that coatings were mechanically strong, chemically stable, and possessed good adhesion strength to the substrate. To further confirm the bone tissue regeneration potential of the coatings, in-vitro bioactivity, cell viability, and cell proliferation tests were done. The obtained data confirmed that CS/Gel/WH composite is highly bioactive and promotes osseointegration. TTO at lower concentrations facilitates cell proliferation but it is toxic at higher concentrations. TTO interacted with the coating matrix via non-covalent interactions that resulted in its slow and sustained release over 28 days. TTO showed antibacterial effect when tested with E. coli bacteria. Hence, in this work, experimental, statistical, physiochemical, and biological analyses are discussed in detail based on the current knowledge of WH coatings and the effect of TTO on cytocompatibility as well as biofilm prevention.
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