临床应用的新鲜骨软骨同种异体移植物的保存条件至关重要，因为它们的目标是：移植含有活软骨细胞的成熟透明软骨，维持其代谢活性，并保持细胞外基质的结构和功能特征。本研究的目的是比较荧光共聚焦显微镜和流式细胞术技术，以评估骨软骨组织中存在的软骨细胞的活力，以确定其有效性，从而确保分析的可重复性和稳健性。为此,将来自人类尸体供体的同种异体骨软骨移植物在补充有抗生素和抗真菌剂的保存培养基中在4°C下保存3周。通过共聚焦荧光显微镜和流式细胞术监测软骨保存3周，确定软骨细胞的细胞活力，第三周获得83.7±2.6%和55.8±7.8%的细胞活力，分别。共聚焦荧光显微镜方法更具优势和准确性，因为它与监测骨软骨移植物保存的实际细胞活力值更好地相关，仅检测与保存方法相关的自然死亡细胞。
The preservation conditions of fresh osteochondral allografts for clinical applications are critical due their objective: to transplant mature hyaline cartilage containing viable chondrocytes, maintaining their metabolic activity and also preserving the structural and functional characteristics of the extracellular matrix. The aim of the present
study was to compare fluorescence confocal microscopy and flow cytometry techniques to evaluate the viability of the chondrocytes present in the osteochondral tissue, in order to determine their effectiveness and thus ensure reproducibility and robustness of the analysis. To this end, osteochondral allografts from human cadaveric donors were preserved at 4 °C for 3 weeks in a preservation medium supplemented with antibiotic and antifungal agents. Cell viability of chondrocytes was determined by monitoring the cartilage for 3 weeks of preservation by confocal fluorescence microscopy and flow cytometry, obtaining cell viabilities of 83.7 ± 2.6% and 55.8 ± 7.8% for week three, respectively. The confocal fluorescence microscopy approach is more advantageous and accurate, as it correlates better with actual cell viability values for monitoring osteochondral graft preservation, detecting only the cells that died a natural death associated with the preservation method.
