In vivo biocompatibility analysis of the recellularized canine tracheal scaffolds with canine epithelial and endothelial progenitor cells

Bioengineered. 2022 Feb;13(2):3551-3565. doi: 10.1080/21655979.2021.2020392.

Abstract

Decellularized extracellular matrix (ECM) has frequently been applied as a biomaterial for tissue engineering purposes. When implanted, their role can be essential for partial trachea replacement in patients that require a viable transplant solution. Acellular canine tracheal scaffolds with preserved ECM structure, flexibility, and proteins were obtained by high pressure vacuum decellularization. Here, we aimed to evaluate the cell adhesion and proliferation of canine tracheal epithelial cells (EpC) and canine yolk sac endothelial progenitor cells (YS) cultivated on canine decellularized tracheal scaffolds and test the in vivo biocompatibility of these recellularized scaffolds implanted in BALB-c nude mice. In order to evaluate the recellularization efficiency, scaffolds were evaluated by scanning electron microscopy (SEM), immunofluorescence, DNA quantification, mycoplasma test, and in vivo biocompatibility. The scaffolds sterility was confirmed, and EpC and YS cells were cultured by 7 and 14 days. We demonstrated by SEM, immunofluorescence, and genomic DNA analyzes cell adhesion to tracheal ECM. Then, recellularized scaffolds were in vivo subcutaneously implanted in mice and after 45 days, the fragments were collected and analyzed by Hematoxylin-Eosin and Gömori Trichrome staining and PCNA, CD4, CD8, and CD68 immunohistochemistry. In vivo results confirmed that the implanted tissue remains preserved and proliferative, and no fibrotic tissue process was observed in animals. Finally, our results showed the recellularization success due the preserved ECM proteins, and that these may be suitable to future preclinical studies applications for partial trachea replacement in tissue engineering.

Keywords: Cartilage; decellularized scaffolds; tissue engineering; tracheal collapse; transplants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Dogs
  • Endothelial Progenitor Cells*
  • Extracellular Matrix
  • Humans
  • Mice
  • Mice, Nude
  • Tissue Engineering / methods
  • Tissue Scaffolds
  • Trachea*

Grants and funding

This work was supported by The São Paulo Research Foundation [FAPESP, grant number Fundação de Amparo à Pesquisa do Estado de São Paulo 2014/50844-3] and CAPES [Coordination for the Improvement of Higher Education Personnel, grant number Coordenação de Aperfeiçoamento de Pessoal de Nível Superior 88887.488264/2020-00] and PROEX: 88882.327806/2014-01.