Analysis of CGF Biomolecules, Structure and Cell Population: Characterization of the Stemness Features of CGF Cells and Osteogenic Potential

• Published in: International journal of molecular sciences 2021-08, Vol.22 (16), p.8867
• Main Authors: Stanca, Eleonora, Calabriso, Nadia, Giannotti, Laura, Nitti, Paola, Damiano, Fabrizio, Stanca, Benedetta Di Chiara, Carluccio, Maria Annunziata, De Benedetto, Giuseppe Egidio, Demitri, Christian, Palermo, Andrea, Ferrante, Franco, Siculella, Luisa, Rochira, Alessio
• Format: Article
• Language: English
• Subjects: Alizarin; Anticoagulants; Biomaterials; Biomedical materials; Biomolecules; Blood platelets; blood-derived biomaterials; Cell differentiation; CGF; Differentiation (biology); Enzyme-linked immunosorbent assay; Fibroblasts; Flow cytometry; growth factor; Growth factors; Immunohistochemistry; Kinetics; Matrix metalloproteinase; Matrix metalloproteinases; Metabolites; Mineralization; mRNA; osteogenic differentiation; Real time; Regenerative medicine; Stem cells; Structural analysis; Surface markers; Tissue engineering; Vascular endothelial growth factor
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22168867

Concentrated Growth Factors (CGF) represent new autologous (blood-derived biomaterial), attracting growing interest in the field of regenerative medicine. In this study, the chemical, structural, and biological characterization of CGF was carried out. CGF molecular characterization was performed by GC/MS to quantify small metabolites and by ELISA to measure growth factors and matrix metalloproteinases (MMPs) release; structural CGF characterization was carried out by SEM analysis and immunohistochemistry; CGF has been cultured, and its primary cells were isolated for the identification of their surface markers by flow cytometry, Western blot, and real-time PCR; finally, the osteogenic differentiation of CGF primary cells was evaluated through matrix mineralization by alizarin red staining and through mRNA quantification of osteogenic differentiation markers by real-time PCR. We found that CGF has a complex inner structure capable of influencing the release of growth factors, metabolites, and cells. These cells, which could regulate the production and release of the CGF growth factors, show stem features and are able to differentiate into osteoblasts producing a mineralized matrix. These data, taken together, highlight interesting new perspectives for the use of CGF in regenerative medicine.