The present study reports the development of a novel film-forming bovine collagenous cream (BCC) based on bovine collagen (BC). In this study, collagen was isolated from bovine forestomach tissue, a novel source, and a cream formulation was prepared using some other bioactive ingredients. The electrophoretic pattern of the BC was found to be similar to type I collagen. The purity of BC was examined by amino acid analysis, which confirmed the presence of atelocollagen. The physicochemical properties of BCC such as rheology, spreadability, and temperature stability were characterized. The antimicrobial activity was examined against Bacillus subtilis, Staphylococcus aureus, and Escherichia coli, and BCC displayed excellent inhibitory effect. In vitro biocompatibility studies using NIH 3T3 fibroblast cells showed enhanced cell viability. FACS analysis revealed the non-toxic nature of BCC toward cells. The cell morphology and proliferation on the BCC matrix was studied using SEM and fluorescence microscopy. The in vivo wound healing efficacy of the BCC as a topical wound dressing was demonstrated on full thickness excision wounds in rat models. The healing profile showed that the BCC significantly enhanced re-epithelialization, collagen deposition, and contraction in the wound healing process. The findings of this study provide a new opportunity for the utilization of the untapped byproducts of the meat industry for valorization. We expect that this kind of topical healing cream could be a potential candidate in wound management and future clinical needs.