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

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 <i<in vitro</i< and experimental animal models p...
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Autor*in:	Serena Duchi [verfasserIn] Stephanie Doyle [verfasserIn] Timon Eekel [verfasserIn] Cathal D. O’Connell [verfasserIn] Cheryl Augustine [verfasserIn] Peter Choong [verfasserIn] Carmine Onofrillo [verfasserIn] Claudia Di Bella [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2019

Schlagwörter:	cartilage osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration

Übergeordnetes Werk:	In: Materials - MDPI AG, 2009, 12(2019), 4, p 640
Übergeordnetes Werk:	volume:12 ; year:2019 ; number:4, p 640

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3390/ma12040640

Katalog-ID:	DOAJ012227919

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allfields	10.3390/ma12040640 doi (DE-627)DOAJ012227919 (DE-599)DOAJ70743ff028194077ab079ac297b34f0b DE-627 ger DE-627 rakwb eng TK1-9971 TA1-2040 QH201-278.5 QC120-168.85 Serena Duchi verfasserin aut Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing. cartilage <i<ex vivo</i< model osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration Technology T Electrical engineering. Electronics. Nuclear engineering Engineering (General). Civil engineering (General) Microscopy Descriptive and experimental mechanics Stephanie Doyle verfasserin aut Timon Eekel verfasserin aut Cathal D. O’Connell verfasserin aut Cheryl Augustine verfasserin aut Peter Choong verfasserin aut Carmine Onofrillo verfasserin aut Claudia Di Bella verfasserin aut In Materials MDPI AG, 2009 12(2019), 4, p 640 (DE-627)595712649 (DE-600)2487261-1 19961944 nnns volume:12 year:2019 number:4, p 640 https://doi.org/10.3390/ma12040640 kostenfrei https://doaj.org/article/70743ff028194077ab079ac297b34f0b kostenfrei https://www.mdpi.com/1996-1944/12/4/640 kostenfrei https://doaj.org/toc/1996-1944 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2119 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2019 4, p 640
spelling	10.3390/ma12040640 doi (DE-627)DOAJ012227919 (DE-599)DOAJ70743ff028194077ab079ac297b34f0b DE-627 ger DE-627 rakwb eng TK1-9971 TA1-2040 QH201-278.5 QC120-168.85 Serena Duchi verfasserin aut Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing. cartilage <i<ex vivo</i< model osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration Technology T Electrical engineering. Electronics. Nuclear engineering Engineering (General). Civil engineering (General) Microscopy Descriptive and experimental mechanics Stephanie Doyle verfasserin aut Timon Eekel verfasserin aut Cathal D. O’Connell verfasserin aut Cheryl Augustine verfasserin aut Peter Choong verfasserin aut Carmine Onofrillo verfasserin aut Claudia Di Bella verfasserin aut In Materials MDPI AG, 2009 12(2019), 4, p 640 (DE-627)595712649 (DE-600)2487261-1 19961944 nnns volume:12 year:2019 number:4, p 640 https://doi.org/10.3390/ma12040640 kostenfrei https://doaj.org/article/70743ff028194077ab079ac297b34f0b kostenfrei https://www.mdpi.com/1996-1944/12/4/640 kostenfrei https://doaj.org/toc/1996-1944 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2119 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2019 4, p 640
allfields_unstemmed	10.3390/ma12040640 doi (DE-627)DOAJ012227919 (DE-599)DOAJ70743ff028194077ab079ac297b34f0b DE-627 ger DE-627 rakwb eng TK1-9971 TA1-2040 QH201-278.5 QC120-168.85 Serena Duchi verfasserin aut Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing. cartilage <i<ex vivo</i< model osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration Technology T Electrical engineering. Electronics. Nuclear engineering Engineering (General). Civil engineering (General) Microscopy Descriptive and experimental mechanics Stephanie Doyle verfasserin aut Timon Eekel verfasserin aut Cathal D. O’Connell verfasserin aut Cheryl Augustine verfasserin aut Peter Choong verfasserin aut Carmine Onofrillo verfasserin aut Claudia Di Bella verfasserin aut In Materials MDPI AG, 2009 12(2019), 4, p 640 (DE-627)595712649 (DE-600)2487261-1 19961944 nnns volume:12 year:2019 number:4, p 640 https://doi.org/10.3390/ma12040640 kostenfrei https://doaj.org/article/70743ff028194077ab079ac297b34f0b kostenfrei https://www.mdpi.com/1996-1944/12/4/640 kostenfrei https://doaj.org/toc/1996-1944 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2119 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2019 4, p 640
allfieldsGer	10.3390/ma12040640 doi (DE-627)DOAJ012227919 (DE-599)DOAJ70743ff028194077ab079ac297b34f0b DE-627 ger DE-627 rakwb eng TK1-9971 TA1-2040 QH201-278.5 QC120-168.85 Serena Duchi verfasserin aut Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing. cartilage <i<ex vivo</i< model osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration Technology T Electrical engineering. Electronics. Nuclear engineering Engineering (General). Civil engineering (General) Microscopy Descriptive and experimental mechanics Stephanie Doyle verfasserin aut Timon Eekel verfasserin aut Cathal D. O’Connell verfasserin aut Cheryl Augustine verfasserin aut Peter Choong verfasserin aut Carmine Onofrillo verfasserin aut Claudia Di Bella verfasserin aut In Materials MDPI AG, 2009 12(2019), 4, p 640 (DE-627)595712649 (DE-600)2487261-1 19961944 nnns volume:12 year:2019 number:4, p 640 https://doi.org/10.3390/ma12040640 kostenfrei https://doaj.org/article/70743ff028194077ab079ac297b34f0b kostenfrei https://www.mdpi.com/1996-1944/12/4/640 kostenfrei https://doaj.org/toc/1996-1944 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2119 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2019 4, p 640
allfieldsSound	10.3390/ma12040640 doi (DE-627)DOAJ012227919 (DE-599)DOAJ70743ff028194077ab079ac297b34f0b DE-627 ger DE-627 rakwb eng TK1-9971 TA1-2040 QH201-278.5 QC120-168.85 Serena Duchi verfasserin aut Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing. cartilage <i<ex vivo</i< model osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration Technology T Electrical engineering. Electronics. Nuclear engineering Engineering (General). Civil engineering (General) Microscopy Descriptive and experimental mechanics Stephanie Doyle verfasserin aut Timon Eekel verfasserin aut Cathal D. O’Connell verfasserin aut Cheryl Augustine verfasserin aut Peter Choong verfasserin aut Carmine Onofrillo verfasserin aut Claudia Di Bella verfasserin aut In Materials MDPI AG, 2009 12(2019), 4, p 640 (DE-627)595712649 (DE-600)2487261-1 19961944 nnns volume:12 year:2019 number:4, p 640 https://doi.org/10.3390/ma12040640 kostenfrei https://doaj.org/article/70743ff028194077ab079ac297b34f0b kostenfrei https://www.mdpi.com/1996-1944/12/4/640 kostenfrei https://doaj.org/toc/1996-1944 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2119 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2019 4, p 640
language	English
source	In Materials 12(2019), 4, p 640 volume:12 year:2019 number:4, p 640
sourceStr	In Materials 12(2019), 4, p 640 volume:12 year:2019 number:4, p 640
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	cartilage <i<ex vivo</i< model osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration Technology T Electrical engineering. Electronics. Nuclear engineering Engineering (General). Civil engineering (General) Microscopy Descriptive and experimental mechanics
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container_title	Materials
authorswithroles_txt_mv	Serena Duchi @@aut@@ Stephanie Doyle @@aut@@ Timon Eekel @@aut@@ Cathal D. O’Connell @@aut@@ Cheryl Augustine @@aut@@ Peter Choong @@aut@@ Carmine Onofrillo @@aut@@ Claudia Di Bella @@aut@@
publishDateDaySort_date	2019-01-01T00:00:00Z
hierarchy_top_id	595712649
id	DOAJ012227919
language_de	englisch
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callnumber-first	T - Technology
author	Serena Duchi
spellingShingle	Serena Duchi misc TK1-9971 misc TA1-2040 misc QH201-278.5 misc QC120-168.85 misc cartilage misc <i<ex vivo</i< model misc osteochondral unit misc hydrogel-based scaffold misc histological procedures misc cartilage regeneration misc Technology misc T misc Electrical engineering. Electronics. Nuclear engineering misc Engineering (General). Civil engineering (General) misc Microscopy misc Descriptive and experimental mechanics Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications
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topic_title	TK1-9971 TA1-2040 QH201-278.5 QC120-168.85 Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications cartilage <i<ex vivo</i< model osteochondral unit hydrogel-based scaffold histological procedures cartilage regeneration
topic	misc TK1-9971 misc TA1-2040 misc QH201-278.5 misc QC120-168.85 misc cartilage misc <i<ex vivo</i< model misc osteochondral unit misc hydrogel-based scaffold misc histological procedures misc cartilage regeneration misc Technology misc T misc Electrical engineering. Electronics. Nuclear engineering misc Engineering (General). Civil engineering (General) misc Microscopy misc Descriptive and experimental mechanics
topic_unstemmed	misc TK1-9971 misc TA1-2040 misc QH201-278.5 misc QC120-168.85 misc cartilage misc <i<ex vivo</i< model misc osteochondral unit misc hydrogel-based scaffold misc histological procedures misc cartilage regeneration misc Technology misc T misc Electrical engineering. Electronics. Nuclear engineering misc Engineering (General). Civil engineering (General) misc Microscopy misc Descriptive and experimental mechanics
topic_browse	misc TK1-9971 misc TA1-2040 misc QH201-278.5 misc QC120-168.85 misc cartilage misc <i<ex vivo</i< model misc osteochondral unit misc hydrogel-based scaffold misc histological procedures misc cartilage regeneration misc Technology misc T misc Electrical engineering. Electronics. Nuclear engineering misc Engineering (General). Civil engineering (General) misc Microscopy misc Descriptive and experimental mechanics
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title	Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications
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title_full	Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications
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author_browse	Serena Duchi Stephanie Doyle Timon Eekel Cathal D. O’Connell Cheryl Augustine Peter Choong Carmine Onofrillo Claudia Di Bella
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title_sort	protocols for culturing and imaging a human <i<ex vivo</i< osteochondral model for cartilage biomanufacturing applications
callnumber	TK1-9971
title_auth	Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications
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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing.
abstractGer	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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing.
abstract_unstemmed	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 <i<in vitro</i< and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. <i<Ex vivo</i< models are of great value for translating <i<in vitro</i< 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 <i<ex vivo</i< 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, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical <i<ex vivo</i< tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for <i<in vivo</i< testing.
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title_short	Protocols for Culturing and Imaging a Human <i<Ex Vivo</i< Osteochondral Model for Cartilage Biomanufacturing Applications
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