A Comparative Study Using Fluorescent Confocal Microscopy and Flow Cytometry to Evaluate Chondrocyte Viability in Human Osteochondral Allografts
López-Chicón, Patricia (Institut de Recerca Biomèdica de Lleida)
Riba-Tietz, Tatiana (Institut de Recerca Biomèdica de Lleida)
Fariñas Barbera, Oscar (Institut de Recerca Biomèdica de Lleida)
Gelber Ghertner, Pablo Eduardo (Institut d'Investigació Biomèdica Sant Pau)
Casaroli-Marano, Ricardo Pedro (Hospital Clínic i Provincial de Barcelona)
Vilarrodona, Anna (Hospital Universitari Vall d'Hebron)
Universitat Autònoma de Barcelona

Date:	2022
Abstract:	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.
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Osteoarthritis ; Osteochondral grafts ; Chondrocytes cell viability ; Graft preservation ; Confocal fluorescence microscopy ; Flow cytometry
Published in:	Bioengineering, Vol. 9 Núm. 10 (october 2022) , p. 515, ISSN 2306-5354

DOI: 10.3390/bioengineering9100515
PMID: 36290483

13 p, 2.7 MB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Recerca Sant Pau
Articles > Research articles
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