The effect of local cryostimulation on osteogenesis during implantation of xenogenic bone matrix: An experimental...

Intensive skin cooling triggers a physiological response that has therapeutic potential for bone injury treatment. However, in the cases of critical-sized bone defects, cryostimulation alone is insufficient. Then this study aimed to evaluate the rate of bone tissue regeneration in the case of the combination of cryostimulation and xenogenic bone matrix implantation to test the hypothesis of healing acceleration. A model of a critical-size defect (2.7 mm in diameter) in the cortical bone of the femoral diaphysis was drilled out in Wistar rats. The defects were filled with xenogenic bone matrix. After cryostimulation carried out twice per week over a 30-day period. Bone regeneration was assessed with micro-CT, histological, immunohistochemical and fluorescence microscopy tests. Micro-CT and histological evaluations revealed enhanced bone regeneration with structural features typical of compact bone, including Haversian canal formation and the presence of cement lines. Morphometric analysis demonstrated a significantly greater relative area of bone regeneration in the experimental group. Immunohistochemistry revealed an increased alkaline phosphatase activity, indicating elevated osteoblastic activity. Fluorescence microscopy confirmed a statistically significant moderate increase (by 10 %) in bone tissue growth rate. Overall, the combined application of cryostimulation and xenogenic bone matrix implantation resulted in complete defect filling and near-complete mineralization of the newly formed bone. These findings demonstrate that cryostimulation enhances bone regeneration when filled xenogenic bone matrix. It offering a promising approach for the treatment of critical-sized bone defects. The results may contribute to the design of novel therapeutic protocols for bone healing, particularly in complicated clinical cases.