Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration.
Extracellular matrix (ECM) is a fundamental component of the heart, guiding vital cellular processes during organ homeostasis. Most cardiovascular diseases lead to a remarkable remodeling of the ECM, accompanied by the formation of a fibrotic tissue that heavily compromises the heart function. Effec...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2022-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0276224 |
| _version_ | 1797989489029677056 |
|---|---|
| author | Immacolata Belviso Anna Maria Sacco Domenico Cozzolino Daria Nurzynska Franca Di Meglio Clotilde Castaldo Veronica Romano |
| author_facet | Immacolata Belviso Anna Maria Sacco Domenico Cozzolino Daria Nurzynska Franca Di Meglio Clotilde Castaldo Veronica Romano |
| author_sort | Immacolata Belviso |
| collection | DOAJ |
| description | Extracellular matrix (ECM) is a fundamental component of the heart, guiding vital cellular processes during organ homeostasis. Most cardiovascular diseases lead to a remarkable remodeling of the ECM, accompanied by the formation of a fibrotic tissue that heavily compromises the heart function. Effective therapies for managing fibrosis and promoting physiological ECM repair are not yet available. The production of a decellularized extracellular matrix (d-ECM) serving as a three-dimensional and bioactive scaffold able to modulate cellular behavior and activities is considered crucial to achieve a successful regeneration. The protocol represents a step-by-step method to obtain a decellularized cardiac matrix through the combination of sodium dodecyl sulphate (SDS) and Triton X-100. Briefly, cardiac samples obtained from left ventricles of explanted, pathological human hearts were dissected and washed to remove residual body fluids. Samples were then snap-frozen and sliced by a cryostat into 350 μm thick sections. The sections obtained were decellularized using a solution containing 1% Triton X-100 and 1% SDS in combination, for 24 hours, until observing the color change from brownish-red to translucent-white. As a result, the protocol shows efficiency in preserving ECM architecture and protein composition during the whole process, suggesting that it is worthwhile, highly reproducible and produces a well- preserved decellularized extracellular matrix from cardiac samples. Notwithstanding, some limitations need to be addressed, such as the risk for microbial contamination and the unpredictable trend of the protocol when applied to decellularize samples other than myocardium, vessels, or skin. These issues require antibiotics mixture supplement during the procedure followed by UV sterilization, and appropriate adjustments for a tissue-specific utilization, respectively. The protocol is intended to produce a cardiac d-ECM for cell settlement, representing the ideal scaffold for tissue engineering purposes. |
| first_indexed | 2024-04-11T08:21:11Z |
| format | Article |
| id | doaj.art-374b5e0626664f5bad0ed21c99f815e2 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-11T08:21:11Z |
| publishDate | 2022-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-374b5e0626664f5bad0ed21c99f815e22022-12-22T04:34:57ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-011710e027622410.1371/journal.pone.0276224Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration.Immacolata BelvisoAnna Maria SaccoDomenico CozzolinoDaria NurzynskaFranca Di MeglioClotilde CastaldoVeronica RomanoExtracellular matrix (ECM) is a fundamental component of the heart, guiding vital cellular processes during organ homeostasis. Most cardiovascular diseases lead to a remarkable remodeling of the ECM, accompanied by the formation of a fibrotic tissue that heavily compromises the heart function. Effective therapies for managing fibrosis and promoting physiological ECM repair are not yet available. The production of a decellularized extracellular matrix (d-ECM) serving as a three-dimensional and bioactive scaffold able to modulate cellular behavior and activities is considered crucial to achieve a successful regeneration. The protocol represents a step-by-step method to obtain a decellularized cardiac matrix through the combination of sodium dodecyl sulphate (SDS) and Triton X-100. Briefly, cardiac samples obtained from left ventricles of explanted, pathological human hearts were dissected and washed to remove residual body fluids. Samples were then snap-frozen and sliced by a cryostat into 350 μm thick sections. The sections obtained were decellularized using a solution containing 1% Triton X-100 and 1% SDS in combination, for 24 hours, until observing the color change from brownish-red to translucent-white. As a result, the protocol shows efficiency in preserving ECM architecture and protein composition during the whole process, suggesting that it is worthwhile, highly reproducible and produces a well- preserved decellularized extracellular matrix from cardiac samples. Notwithstanding, some limitations need to be addressed, such as the risk for microbial contamination and the unpredictable trend of the protocol when applied to decellularize samples other than myocardium, vessels, or skin. These issues require antibiotics mixture supplement during the procedure followed by UV sterilization, and appropriate adjustments for a tissue-specific utilization, respectively. The protocol is intended to produce a cardiac d-ECM for cell settlement, representing the ideal scaffold for tissue engineering purposes.https://doi.org/10.1371/journal.pone.0276224 |
| spellingShingle | Immacolata Belviso Anna Maria Sacco Domenico Cozzolino Daria Nurzynska Franca Di Meglio Clotilde Castaldo Veronica Romano Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration. PLoS ONE |
| title | Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration. |
| title_full | Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration. |
| title_fullStr | Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration. |
| title_full_unstemmed | Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration. |
| title_short | Cardiac-derived extracellular matrix: A decellularization protocol for heart regeneration. |
| title_sort | cardiac derived extracellular matrix a decellularization protocol for heart regeneration |
| url | https://doi.org/10.1371/journal.pone.0276224 |
| work_keys_str_mv | AT immacolatabelviso cardiacderivedextracellularmatrixadecellularizationprotocolforheartregeneration AT annamariasacco cardiacderivedextracellularmatrixadecellularizationprotocolforheartregeneration AT domenicocozzolino cardiacderivedextracellularmatrixadecellularizationprotocolforheartregeneration AT darianurzynska cardiacderivedextracellularmatrixadecellularizationprotocolforheartregeneration AT francadimeglio cardiacderivedextracellularmatrixadecellularizationprotocolforheartregeneration AT clotildecastaldo cardiacderivedextracellularmatrixadecellularizationprotocolforheartregeneration AT veronicaromano cardiacderivedextracellularmatrixadecellularizationprotocolforheartregeneration |