A comparative study using fluorescent confocal microscopy and flow cytometry to evaluate chondrocyte viability in human osteochondral allografts.

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 t...

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Detalles Bibliográficos
Autores: López Chicón, Patrícia, Riba Tietz, Tatiana, Fariñas, Óscar, Gelber, Pablo Eduardo, Casaroli Marano, Ricardo Pedro, Vilarrodona, Anna
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/194588
Acceso en línea:https://hdl.handle.net/2445/194588
Access Level:acceso abierto
Palabra clave:Microscòpia confocal
Microscòpia de fluorescència
Citometria de fluxe
Rebuig (Biologia)
Artrosi
Confocal microscopy
Fluorescence microscopy
Flow cytometry
Graft rejection
Osteoarthritis
Descripción
Sumario: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.