Portal de investigação
Pre-selection of fibroblast subsets prompts prevascularization of tissue engineered skin analogues
Data de publicação:
Data Ahead of Print:
Autores da FMUP
Participantes de fora da FMUP
- Moreira, HR
- Cerqueira, MT
- da Silva, LP
- Pires, J
- Jarnalo, M
- Reis, RL
- Marques, AP
Unidades de investigação
Abstract
The papillary and reticular dermis harbors phenotypically distinct fibroblasts, whose functions such as maintenance of skin's microvasculature are also distinct. Thus, we hypothesized that pre-selection of the subpopulations of fibroblasts would benefit the generation of skin tissue engineered (TE) constructs, promoting their prevascularization in vitro. We first isolated papillary and reticular fibroblasts using fluorescence-activated cell sorting and studied the effect of their secretome and extracellular matrix (ECM) on human dermal microvascular endothelial cell (hDMEC) organization. Subsequently, we developed a bilayered 3D polymeric structure with distinct layer-associated features to house the subpopulations of fibroblasts, to generate a skin analogue. Both papillary and reticular fibroblasts were able to stimulate capillary-like network formation in a Matrigel assay. However, the secretome of the two subpopulations was substantially different, being enriched in VEGF, IGF-1, and Angio-1 in the case of papillary fibroblasts and in HGF and FGF-2 for the reticular subset. In addition, the fibroblast subpopulations deposited varied levels of ECM proteins, more collagen I and laminin was produced by the reticular subset, but these differences did not impact hDMEC organization. Vessel-like structures with lumens were observed earlier in the 3D skin analogue prepared with the sorted fibroblasts, although ECM deposition was not affected by the cell's pre-selection. Moreover, a more differentiated epidermal layer was obtained in the skin analogue formed by the sorted fibroblasts, confirming that its whole structure was not affected. Overall, we provide evidence that pre-selection of papillary and reticular fibroblasts is relevant for promoting the in vitro prevascularization of skin TE constructs.
Dados da publicação
- ISSN/ISSNe:
- 2047-4849, 2047-4830
- Tipo:
- Article
- Páginas:
- 5287-5300
- DOI:
- 10.1039/d2bm02022j
- Link para outro recurso:
- www.scopus.com
Biomaterials Science Royal Society of Chemistry
Citações Recebidas na Scopus: 2
Documentos
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Filiações
Keywords
- Cells, Cultured; Collagen Type I; Dermis; Epidermis; Fibroblasts; Humans; Skin; Cell culture; Endothelial cells; Tissue; collagen type 1; fibroblast growth factor 2; laminin; matrigel; scatter factor; scleroprotein; somatomedin C; vasculotropin; collagen type 1; Endothelial cell organizations; Extracellular matrices; Fluorescence activated cell sorting; In-vitro; Microvascular endothelial cells; Pre-selection; Secretome; Skin microvasculature; Skin tissue; Tissue engineered constructs; Article; controlled study; dermal microvascular endothelial cell; extracellular matrix; fibroblast; fluorescence activated cell sorting; gene expression; human; human cell; in vitro study; secretome; cell culture; dermis; epidermis; fibroblast; metabolism; skin; Fibroblasts
Financiamento
Proyectos asociados
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Investigador Principal: Ricardo José Moreira Horta Oliveira
Estudo Clínico Académico . 2021
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Estudo Clínico Académico . 2022
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Citar a publicação
Moreira HR,Cerqueira MT,da LP,Pires J,Jarnalo M,Horta R,Reis RL,Marques AP. Pre-selection of fibroblast subsets prompts prevascularization of tissue engineered skin analogues. Biomater. Sci. 2023. 11. (15):p. 5287-5300. IF:6,600. (2).
