Purpose: The purpose of this study was to evaluate the cell viability, surface characteristics, and osteoconductive potential of ovine bone graft modified with L-glutamine compared to commercial bone graft. Materials and methods: Hydroxyapatite was isolated from the femur of a goat by sintering at 360°C for 3 hours (group A). L-glutamine (HIMEDIA) was introduced into the hydroxyapatite-sintered material and named as group B. Commercially available bone graft (Bio-Oss, Geistlich) serves as the control (group C). The functional groups of the samples were characterized using a Fourier transform infrared spectrophotometer (FTIR), employing the potassium bromide pellet method. The phase composition of the groups was analyzed using X-ray diffractometer. The surface morphology and pore size of the samples were examined with a scanning electron microscope (SEM). Bone formation assay (BFA) was done to check for the optical density (OD) of samples, hence portraying the osteoconductive potential of the sample. The 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay technique was utilized to assess cell viability. Qualitative analysis was done using X-ray diffraction (XRD), FTIR, and SEM analysis. One-way analysis of variance (ANOVA) was done for the intergroup comparison of the BFA and MTT assays. A post hoc Tukey test was done to find the difference between the groups. Results: FTIR demonstrated the presence of specific functional groups in bone graft characterized with L-glutamine, like phosphate and carbonate whose presence indicates the presence of hydroxyapatite. XRD analysis highlighted the presence of hydroxyapatite in the sintered ovine bone samples confirming the suitability of these samples for a wide array of biomedical applications. Analysis of images obtained with SEM demonstrated a tightly bonded structure characterized by densely interwoven particles and distinct bioactive surface layers on the exterior of the bone graft characterized with L-glutamine. MTT assay demonstrated a significant difference among the three groups (p = 0.001), wherein group B demonstrated the highest values of cell viability. Findings for BFA indicate significant differences (p = 0.001) between the three groups, with group B demonstrating the highest values of bone formation assay, the other two groups, the ovine graft (group A) and control (group C) do not differ significantly (p = 0.911). Conclusion: L-glutamine positively affects natural bone graft growth, enhancing understanding of bone regulation. All characterizations demonstrated optimum qualities of the ovine bone graft characterized with L-glutamine, proving to be equivalent to commercially available bone graft, and could be made more economically viable for use in most clinical practices.