Improving the mechanical and biological functions of cell sheet constructs: The interplay of human-derived periodontal ligament stem cells, endothelial cells and plasma rich in growth factors - 27/04/24
, María Troya a, b, Mar Zalduendo a, b, Roberto Tierno a, b, Mohammad H. Alkhraisat a, b, Nerea Osinalde c, Asier Fullaondo b, d, Ana M. Zubiaga b, dAbstract |
Objective |
The aim of this study was to produce and characterize triple-layered cell sheet constructs with varying cell compositions combined or not with the fibrin membrane scaffold obtained by the technology of Plasma Rich in Growth Factors (mPRGF).
Materials and methods |
Human primary cultures of periodontal ligament stem cells (hPDLSCs) were isolated, and their stemness nature was evaluated. Three types of triple-layered composite constructs were generated, composed solely of hPDLSCs or combined with human umbilical vein endothelial cells (HUVECs), either as a sandwiched endothelial layer or as coculture sheets of both cell phenotypes. These three triple-layered constructs were also manufactured using mPRGF as cell sheets’ support. Necrosis, glucose consumption, secretion of extracellular matrix proteins and synthesis of proangiogenic factors were determined. Histological evaluations and proteomic analyses were also performed.
Results |
The inclusion of HUVECs did not clearly improve the properties of the multilayered constructs and yet hindered their optimal conformation. The presence of mPRGF prevented the shrinkage of cell sheets, stimulated the metabolic activity and increased the matrix synthesis. At the proteome level, mPRGF conferred a dramatic advantage to the hPDLSC constructs in their ability to provide a suitable environment for tissue regeneration by inducing the expression of proteins necessary for bone morphogenesis and cellular proliferation.
Conclusions |
hPDLSCs’ triple-layer construct onto mPRGF emerges as the optimal structure for its use in regenerative therapeutics.
Clinical relevance |
These results suggest the suitability of mPRGF as a promising tool to support cell sheet formation by improving their handling and biological functions.
Le texte complet de cet article est disponible en PDF.Graphical abstract |
Schematic representation of the research study. A) Isolation and characterisation of hPDLSCs. B) Preparation of PRGF formulations. C) Fabrication of the different triple-layered cell constructs and assays performed. Created with BioRender.com.
Schematic representation of the research study. A) Isolation and characterisation of hPDLSCs. B) Preparation of PRGF formulations. C) Fabrication of the different triple-layered cell constructs and assays performed. Created with BioRender.com.Le texte complet de cet article est disponible en PDF.
HIGHLIGHTS |
• | CST avoids proteolytic enzymes for cell detachment, preserving efficiently ECM components. |
• | The weak mechanical properties of cell sheets constitute an important limitation. |
• | mPRGF prevents shrinkage, facilitates handling and provides biological cues. |
• | The inclusion of HUVECs hindered the optimal formation of cell sheets constructs. |
• | hPDLSCs’ tri-layer construct onto mPRGF emerges as the optimal option. |
Keywords : Plasma Rich in Growth Factors, Platelet-rich fibrin, Cell Sheet Technology, Multilayered constructs, Tissue regeneration
Plan
Vol 174
Article 116599- mai 2024 Retour au numéro
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