Cryogenic, but not hypothermic, preservation disrupts the extracellular matrix of cell sheets

Data de publicação: 01/01/2025

Autores da FMUP

Ricardo José Moreira Horta Oliveira

Autor

Participantes de fora da FMUP

Freitas-Ribeiro S.
Carvalho A.F.
Rodrigues D.B.
Martins L.
Pires R.A.
Mendes V.M.
Manadas B.
Jarnalo M.
Reis R.L.
Pirraco R.P.

Unidades de investigação

Cirurgia e Fisiologia

Abstract

Cell sheet (CS)-based approaches hold significant potential for tissue regeneration, relying on the extracellular matrix (ECM) for success. Like in native tissues, the ECM provides structural and biochemical support for cellular homeostasis and function. Effective preservation strategies that maintain ECM integrity are critical to enhance the therapeutic potential of CS-based approaches. While cryogenic and hypothermic preservation methods offer potential solutions, their impact on CS ECM structure is not fully understood. Therefore, a comprehensive analysis of the ECM of hASCs CS following cryogenic and hypothermic preservation for 3 and 7 days, was conducted. Although proteomic analysis indicated that cryopreservation had no significant effect on the overall composition of the ECM, it induced significant ECM structural alterations, particularly disrupting collagen organization, which was not observed following hypothermic preservation. These structural changes were accompanied by alterations in mechanical properties, including a reduction in elastic modulus. In contrast, hypothermic preservation maintained ECM integrity and mechanical properties similar to the control. The notable ECM structural changes following cryogenic preservation can potentially impact cellular behavior, including adhesion, proliferation, and differentiation, thereby affecting the efficacy of CS therapies in vivo. This suggests that hypothermia may offer a promising alternative to cryopreservation for preserving CS integrity and functionality. © 2024 The Authors

Dados da publicação

ISSN/ISSNe:: 2452-199X, 2452-199X
Tipo:: Article
Páginas:: 301-310
DOI:: 10.1016/j.bioactmat.2024.12.019
PubMed:: 39811467
Link para outro recurso:: www.scopus.com

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Freitas-Ribeiro S.:: 3B's Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Rua Ave 1, Edifício 1 (Sede), 4805-694 Barco, Guimarães, Portugal

ICVS/3B's–PT Government Associate Laboratory, Braga/Guimarães, Portugal
Carvalho A.F.:: 3B's Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Rua Ave 1, Edifício 1 (Sede), 4805-694 Barco, Guimarães, Portugal

ICVS/3B's–PT Government Associate Laboratory, Braga/Guimarães, Portugal

Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade Do Porto, Porto, Portugal
Rodrigues D.B.:: 3B's Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Rua Ave 1, Edifício 1 (Sede), 4805-694 Barco, Guimarães, Portugal

ICVS/3B's–PT Government Associate Laboratory, Braga/Guimarães, Portugal
Martins L.:: 3B's Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Rua Ave 1, Edifício 1 (Sede), 4805-694 Barco, Guimarães, Portugal

ICVS/3B's–PT Government Associate Laboratory, Braga/Guimarães, Portugal
Pires R.A.:: 3B's Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Rua Ave 1, Edifício 1 (Sede), 4805-694 Barco, Guimarães, Portugal

ICVS/3B's–PT Government Associate Laboratory, Braga/Guimarães, Portugal
Mendes V.M.:: CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal
Manadas B.:: CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal

Institute for Interdisciplinary Research, University of Coimbra, Coimbra, Portugal
Jarnalo M.:: Department of Plastic and Reconstructive Surgery, and Burn Unity, Centro Hospitalar de São João, Porto, Portugal

Faculty of Medicine - University of Porto, Portugal
Horta R.:: Department of Plastic and Reconstructive Surgery, and Burn Unity, Centro Hospitalar de São João, Porto, Portugal

Faculty of Medicine - University of Porto, Portugal
Reis R.L.:: 3B's Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Rua Ave 1, Edifício 1 (Sede), 4805-694 Barco, Guimarães, Portugal

ICVS/3B's–PT Government Associate Laboratory, Braga/Guimarães, Portugal
Pirraco R.P.:: 3B's Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Rua Ave 1, Edifício 1 (Sede), 4805-694 Barco, Guimarães, Portugal

ICVS/3B's–PT Government Associate Laboratory, Braga/Guimarães, Portugal

Keywords

Article; atomic force microscopy; biopreservation; cell adhesion; cell differentiation; cell engineering; cell function; cell proliferation; cell sheet; cellular homeostasis; controlled study; cryogenic preservation; cryopreservation; differentially expressed protein analysis; erythrocyte; extracellular matrix; histology; homeostasis; human; human cell; hypothermia; immunohistochemistry; in vivo study; Masson staining; preservation; protein analysis; proteomics