Research on the synthesis of multifunctional metal-based nanoparticles using natural extracts is expanding quickly in the biomedical field. Zinc oxide nanoparticles (ZnO-NPs) are well-known bioactive inorganic nanoparticles that have been extensively investigated in regenerative medicine. Herein, ZnO-NPs were prepared using water soluble zinc chloride salt with sodium hydroxide in the presence of Triethanolamine (TEA), as a surfactant agent. ZnO-NPs were functionalized with the Oleo gum resin (BSE) using an in-situ loading approach. The morphology and structure of the as-prepared materials were characterized using TEM, XRD, and Ȥ-potential & DLS. In addition, the cell viability of normal human skin fibroblasts (HSF) after its exposure to varying concentrations of the as-prepared ZnO-NPs and BSE-loaded ZnO-NPs. Although, the XRD data indicates the formation of Zincite phase in the presence/absence of BSE. The TEM results show that ZnO-NPs changed from a rod-like structure with an average particle size 172±29nm in width and 40±12nm in length to an irregular spherical NPs with an average particle size 49±8nm for 10BSE-ZnO-NPs and 15±3nm for 20BSE-ZnO-NPs. The MTT test proves that 20BSE-ZnO-NPs enhance cell growth and show reduced toxicity against HSF cells compared to 10BSE-ZnO-NPs. the obtained results suggest that the ZNO-NPs containing BSE provide a better stimulatory response against Human fibroblast cells, which confirm that the BSE loaded-ZnO-NPs may be utilized in skin tissue healing application.