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Home / Equine Research Bank / J Tissue Eng / Equine lung decellularization: a potential approach for in v...
Journal of tissue engineering2018; 9; 2041731418810164; doi: 10.1177/2041731418810164

Equine lung decellularization: a potential approach for in vitro modeling the role of the extracellular matrix in asthma.

Abstract: Contrary to conventional research animals, horses naturally develop asthma, a disease in which the extracellular matrix of the lung plays a significant role. Hence, the horse lung extracellular matrix appears to be an ideal candidate model for in vitro studying the mechanisms and potential treatments for asthma. However, so far, such model to study cell-extracellular matrix interactions in asthma has not been developed. The aim of this study was to establish a protocol for equine lung decellularization that maintains the architecture of the extracellular matrix and could be used in the future as an in vitro model for therapeutic treatment in asthma. For this the equine lungs were decellularized by sodium dodecyl sulfate detergent perfusion at constant gravitational pressure of 30 cmHO. Lung scaffolds were assessed by immunohistochemistry (collagen I, III, IV, laminin, and fibronectin), scanning electron microscopy, and DNA quantification. Their mechanical property was assessed by measuring lung compliance using the super-syringe technique. The optimized protocol of lung equine decellularization was effective to remove cells (19.8 ng/mg) and to preserve collagen I, III, IV, laminin, and fibronectin. Moreover, scanning electron microscopy analysis demonstrated maintained microscopic lung structures. The decellularized lungs presented lower compliance compared to native lung. In conclusion we described a reproducible decellularization protocol that can produce an acellular equine lung feasible for the future development of novel treatment strategies in asthma.
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Publication Date: 2018-11-12 PubMed ID: 30450188PubMed Central: PMC6236489DOI: 10.1177/2041731418810164Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research explores a novel method to study the underlying mechanisms and potential treatments for asthma using the extracellular matrix of a horse’s lung. The aim of the study was to develop a protocol that can effectively remove cells while keeping the structure of the lung scaffold intact, in order for its use as a model in future asthma treatments.

Approach to the Study

  • The researchers sought to provide a thorough investigation of the extracellular matrix (ECM) in the lung scaffolds of horses, given the fact that horses, unlike many conventional research animals, naturally develop asthma. The ECM holds an important role in the disease’s development, making it a suitable model to test possible treatments for asthma.

Decellularization Process

  • The team developed a protocol for equine lung decellularization, which maintains the ECM architecture. This involves removing all the cells from the organ, leaving only the ECM behind, which can then be used for further research.
  • This process was conducted using sodium dodecyl sulfate detergent, perfused at a constant gravitational pressure of 30 cmHO into the equine lungs. This is a common detergent used in decellularization processes because of its ability to disrupt and solubilize cell membranes.

Scaffold Assessment

  • After the decellularization, the structural and mechanical properties of the lung scaffolds were evaluated using a variety of tools including immunohistochemistry, scanning electron microscopy, and DNA quantification.
  • With immunohistochemistry, the researchers analyzed the quantity and distribution of collagen I, III, IV, laminin, and fibronectin in the lung tissue. These elements are all crucial components of the ECM.
  • Scanning electron microscopy was employed to assess microscopic changes in the lung structure after the decellularization process.
  • DNA quantification was performed to verify whether all cells were effectively removed. The test revealed successful removal of cells, indicated by a DNA residue of 19.8 ng/mg.
  • Lastly, the mechanical property of the ECM scaffolds was evaluated by measuring lung compliance using the super-syringe technique. This method involves inflating and then deflating the lung while recording pressure changes.

Outcomes

  • The decellularization process was successful in preserving the ECM scaffold, including its collagen, laminin and fibronectin components. Electron microscopy confirmed the maintenance of microscopic structures within the lung.
  • The study’s findings reveal that the decellularized lungs, though exhibited lower compliance compared to the native lung tissue, have the potential to be used for future in vitro study models in asthma treatment.

Cite This Article

APA
da Palma RK, Fratini P, Schiavo Matias GS, Cereta AD, Guimarães LL, Anunciação ARA, de Oliveira LVF, Farre R, Miglino MA. (2018). Equine lung decellularization: a potential approach for in vitro modeling the role of the extracellular matrix in asthma. J Tissue Eng, 9, 2041731418810164. https://doi.org/10.1177/2041731418810164

Publication

Journal of tissue engineering
ISSN: 2041-7314
NlmUniqueID: 101550131
Country: England
Language: English
Volume: 9
Pages: 2041731418810164
PII: 2041731418810164

Researcher Affiliations

da Palma, Renata Kelly
  • Post Graduate Program in Science of Rehabilitation, University Nove de Julho (UNINOVE), São Paulo, Brazil.
  • Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Fratini, Paula
  • Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Schiavo Matias, Gustavo Sá
  • Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Cereta, Andressa Daronco
  • Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Guimarães, Leticia Lopes
  • Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Anunciação, Adriana Raquel de Almeida
  • Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
de Oliveira, Luis Vicente Franco
  • Medical School, University Center of Anápolis-UniEVANGELICA, Anápolis, Brazil.
Farre, Ramon
  • Unitat Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, Spain.
  • CIBER de Enfermedades Respiratorias, Madrid, Spain.
  • Institut d'Investigacions Biomèdiques August Pi i Sunyer, Barcelona, Spain.
Miglino, Maria Angelica
  • Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.

Conflict of Interest Statement

Declaration of conflicting interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

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