Protocols for Culturing and Imaging a Human Ex Vivo Osteochondral Model for Cartilage Biomanufacturing Applications

Materials
Serena DuchiClaudia Di Bella

Abstract

Cartilage defects and diseases remain major clinical issues in orthopaedics. Biomanufacturing is now a tangible option for the delivery of bioscaffolds capable of regenerating the deficient cartilage tissue. However, several limitations of in vitro and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. Ex vivo models are of great value for translating in vitro tissue engineered approaches into clinically relevant conditions. Our aim is to obtain a viable human osteochondral (OC) model to test hydrogel-based materials for cartilage repair. Here we describe a detailed step-by-step framework for the generation of human OC plugs, their culture in a perfusion device and the processing procedures for histological and advanced microscopy imaging. Our ex vivo OC model fulfils the following requirements: the model is metabolically stable for a relevant culture period of 4 weeks in a perfusion bioreactor, the processing procedures allowed for the analysis of 3 different tissues or materials (cartilage, bone and hydrogel) without compromising their integrity. We determined a protocol and the settings for a non-linear microscopy technique on label free sections. Furthermore...Continue Reading

References

Nov 1, 1961·The Journal of Bone and Joint Surgery. British Volume·R E OUTERBRIDGE
Nov 5, 2003·Nature Biotechnology·Warren R ZipfelWatt W Webb
Sep 30, 2004·Biophysical Journal·Aikaterini ZoumiBruce J Tromberg
Dec 25, 2008·Regenerative Medicine·Martin J StoddartMauro Alini
Apr 16, 2010·Annals of the New York Academy of Sciences·Mary B Goldring, Steven R Goldring
Aug 20, 2011·The Knee·Benedikt L ProffenMartha M Murray
Aug 31, 2011·Tissue Engineering. Part C, Methods·Marloes L de Vries-van MelleGerjo J V M van Osch
Nov 1, 2009·Sports Health·Alice J Sophia FoxScott A Rodeo
Feb 26, 2014·Stem Cell Research & Therapy·Thomas P LozitoRocky S Tuan
Apr 4, 2014·Bone & Joint Research·J L CookJ P Stannard
Dec 3, 2014·Microscopy Research and Technique·Vitor Aparecido CastaniaHelton Luiz Aparecido Defino
Sep 1, 2015·Frontiers in Surgery·Claudia Di BellaPeter F M Choong
Sep 13, 2016·Biotechnology and Bioengineering·Marta OndrésikRui L Reis
Nov 17, 2016·Journal of Translational Medicine·Christoph BartzJeske J Smink
Feb 1, 2017·European Cells & Materials·S L VegaJ A Burdick
Jun 18, 2017·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Wilhelmina Margaretha GroenJos Malda
Jul 21, 2017·Scientific Reports·Serena DuchiPeter F M Choong
Aug 15, 2017·Advanced Healthcare Materials·Andrew C DalyDaniel J Kelly
Sep 25, 2017·Tissue Engineering. Part C, Methods·Jorge A Piedrahita, J Koudy Williams
Feb 16, 2018·Biomedical Microdevices·Derek A NicholsRiccardo Gottardi
Mar 14, 2018·Clinical Orthopaedics and Related Research·Casey Slattery, Christopher Y Kweon
Apr 25, 2018·Disease Models & Mechanisms·Jacqui Anne McGovernDietmar Werner Hutmacher
May 16, 2018·Stem Cells International·Víctor CarrielMiguel Alaminos
Sep 14, 2015·ACS Biomaterials Science & Engineering·C A VilelaR L Reis

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Citations

Jun 28, 2020·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Meike W A KleuskensKeita Ito
Dec 15, 2020·Tissue Engineering. Part B, Reviews·Anna TrengoveAndrea J O'Connor
Jan 12, 2021·Current Osteoporosis Reports·E E A CramerS Hofmann
Jan 13, 2022·Tissue Engineering. Part C, Methods·Meike W A KleuskensKeita Ito

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biopsies
imaging techniques

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