Effects of disinfection methods and cryopreservation solution formulations containing BMSCs on the cell viability of autologous bone tissue

Background Autologous bone transplantation is an important method for cranial bone repair and reconstruction. This study systematically investigated the effects of different cryopreservation strategies and disinfection methods on autologous bone cells. Methods Bone marrow mesenchymal stem cells (BMSCs) and collagen were added to the basic cryopreservation solution to evaluate the preservation effect. Peracetic acid-UV (PX-UV), supercritical CO₂ (SC-CO₂) and low-temperature plasma (cold atmospheric plasma, CAP) were used to treat skull specimens. Cell viability was detected by CCK-8, cell apoptosis was evaluated by TUNEL staining. RT-qPCR was performed to measure the expression of osteogenic-related genes, including Runt-related transcription factor 2 (Runx2), osteocalcin (OCN), type I collagen (COL1), osteopontin (OPN), and alkaline phosphatase (ALP). Results Adding BMSCs and collagen to the basic cryoprotectant solution significantly increased the survival rate of cryopreserved bone cells ( P < 0.05), inhibited cell apoptosis, and upregulated the expression of osteogenic-related genes. In the disinfection experiment, there were no significant differences between the PX-UV disinfection group and the control group in terms of cell viability, apoptosis rate, and expression of osteogenic genes ( P > 0.05). Conclusion Adding BMSCs and collagen to the basic cryoprotectant solution is expected to improve the preservation effect of autologous bone tissue cells. The PX-UV disinfection has a relatively small impact on the cell survival rate and osteogenic properties of autologous bone cells.