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.
Introduction
Cell-based therapies, regenerative medicine, and tissue engineering have rapidly evolved in recent years. Tissue engineering combines the use of cells, scaffolds, and biological signals to restore the functionality of damaged tissues. However, there are several drawbacks associated with these traditional approaches, such as inflammation, autoimmunity, poor nutrient diffusion, and uneven cell distribution. Cell Sheet Technology (CST) has been proposed as an alternative to overcome these limitations. This technology has been applied in the repair of various tissues, including the heart, cornea, lung, and cartilage. Cell sheets preserve cell-cell interactions and the extracellular matrix (ECM), essential for cellular functions, and offer numerous advantages over conventional regenerative therapies, such as a higher rate of cell survival and biocompatibility.
For a successful tissue engineering approach based on cell sheet technology, variables such as the source of cells and the availability of nutrients must be considered. Mesenchymal stem cells (MSCs) are suitable candidates due to their high expansion rate and differentiation potential. Human periodontal ligament stem cells (hPDLSCs) are an accessible source of stem cells with similar properties. However, the lack of blood vessels in engineered tissues can hinder cell integration and nutrient diffusion. Combining cell sheets with fibrin membranes obtained through Plasma Rich in Growth Factors (PRGF) technology could overcome these physical constraints, providing physical support and biological signals that regulate cell fate and behaviour. In clinical studies, PRGF fibrin membranes have been shown to accelerate ocular surface regeneration and reduce inflammation and fibrosis.
The objective of this study was to produce triple-layer cell sheet constructs with different cellular compositions and analyse their properties in the presence or absence of a fibrin scaffold.
The role of integrin alpha 2 in this study
Integrin alpha 2 is a transmembrane protein that plays a key role in cell adhesion, migration and cell signaling. In this study, its expression was evaluated in triple-layered cell sheet constructs composed of human periodontal ligament stem cells (hPDLSCs) and human umbilical vein endothelial cells (HUVECs). Integrin alpha 2 is essential for the interaction of hPDLSCs with the extracellular matrix, facilitating the formation of a suitable microenvironment for tissue regeneration.
The inclusion of integrin alpha 2 in the constructs was associated with an increase in metabolic activity and extracellular matrix synthesis. These effects were further enhanced with the use of the fibrin membrane obtained through Plasma Rich in Growth Factors (PRGF) technology, which prevented the contraction of the cell sheets and stimulated cell proliferation. Therefore, integrin alpha 2 is fundamental in improving the mechanical and biological properties of cell sheet constructs, promoting a favorable environment for tissue regeneration.
Kits ELISA for the detection of integrin alpha 2
To detect and quantify integrin alpha 2 in the culture media, ELISA kits were used. These kits allow for precise quantification of specific proteins through the use of antibodies that selectively bind to integrin alpha 2. This method is highly sensitive and specific, making it ideal for measuring low levels of proteins in complex biological samples. The ELISA kit used was marketed by Abyntek Research Reagents, with reference ABK1-E4756. This kit enabled accurate measurement of integrin alpha 2 levels, providing crucial data on its role in cell interaction and extracellular matrix formation.
The Abyntek Research Reagents product line is part of Abyntek Biopharma. For over 15 years, Abyntek Biopharma has guided numerous investigations, helping to select the optimal biological reagent to achieve desired results in biomarker analysis. In addition to sharing their knowledge and experience to support these investigations, Abyntek offers an extensive range of high-quality products through their Abyntek Research Reagents line, specifically created to meet the needs of researchers involved in such studies.
Conclusion
The study concludes that triple-layered cell sheet constructs of hPDLSCs on the mPRGF fibrin membrane emerge as the optimal structure for use in regenerative therapies. The inclusion of HUVECs did not significantly enhance the properties of the constructs and actually hindered their optimal formation. The presence of mPRGF prevented the contraction of the cell sheets, stimulated metabolic activity, and increased extracellular matrix synthesis, providing a suitable environment for tissue regeneration by inducing the expression of proteins necessary for bone morphogenesis and cell proliferation.
These results were published in the journal Biomedicine & Pharmacotherapy on April 18, 2024.
