Effects of composite films of silk fibroin and graphene oxide on the proliferation, cell viability and mesenchymal phenotype of periodontal ligament stem cells

• Published in: Journal of materials science. Materials in medicine Vol. 25; no. 12; pp. 2731 - 2741
• Main Authors: Rodríguez-Lozano, F. J., García-Bernal, D., Aznar-Cervantes, S., Ros-Roca, M. A., Algueró, M. C., Atucha, N. M., Lozano-García, A. A., Moraleda, J. M., Cenis, J. L.
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.12.2014; Springer; Springer Nature B.V
• Subjects: Biological and medical sciences; Biomaterials; Biomedical Engineering and Bioengineering; Biomedical materials; Biotechnology; Cell Proliferation - physiology; Cell Survival - physiology; Cells, Cultured; Ceramics; Chemistry and Materials Science; Composite materials; Composites; Culture; Equipment Design; Equipment Failure Analysis; Fibroins - chemistry; Flow cytometry; Fundamental and applied biological sciences. Psychology; Genotype & phenotype; Glass; Graphene; Graphite - chemistry; Head and neck surgery. Maxillofacial surgery. Dental surgery. Orthodontics; Health. Pharmaceutical industry; Humans; Industrial applications and implications. Economical aspects; Materials Science; Maxillofacial surgery. Dental surgery. Orthodontics; Medical sciences; Membranes, Artificial; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - physiology; Miscellaneous; Natural Materials; Oxides - chemistry; Periodontal Ligament - cytology; Periodontal Ligament - physiology; Polymer Sciences; Regenerative; Regenerative Medicine/Tissue Engineering; Stem cells; Surfaces and Interfaces; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Technology. Biomaterials. Equipments; Thin Films; Tissue engineering; Tissue Engineering - instrumentation; Tissue Engineering - methods; Tissue Scaffolds; Viability; Composite Film; Silk Fibroin; Mesenchymal Stem Cell; Graphene Oxide; Mesenchymal Phenotype; Composite film; Human; Cell proliferation; Cell survival; Tissue engineering; Stem cell; Dentistry; Periodontal ligament; Phenotype; Silk; Natural fiber; Animal fiber; Animal protein; Fibroin; Biomedical engineering
• ISSN: 0957-4530; 1573-4838
• DOI: 10.1007/s10856-014-5293-2

In regenerative dentistry, stem cell-based therapy often requires a scaffold to deliver cells and/or growth factors to the injured site. Graphene oxide (GO) and silk fibroin (SF) are promising biomaterials for tissue engineering as they are both non toxic and promote cell proliferation. On the other hand, periodontal ligament stem cells (PDLSCs) are mesenchymal stem cells readily accessible with a promising use in cell therapy. The purpose of this study was to investigate the effects of composite films of GO, SF and GO combined with fibroin in the mesenchymal phenotype, viability, adhesion and proliferation rate of PDLSCs. PDLSCs obtained from healthy extracted teeth were cultured on GO, SF or combination of GO and SF films up to 10 days. Adhesion level of PDSCs on the different biomaterials were evaluated after 12 h of culture, whereas proliferation rate of cells was assessed using the MTT assay. Level of apoptosis was determined using Annexin-V and 7-AAD and mesenchymal markers expression of PDLSCs were analyzed by flow cytometry. At day 7 of culture, MTT experiments showed a high rate of proliferation of PDLSCs growing on GO films compared to the other tested biomaterials, although it was slightly lower than in plastic (control). However PDLSCs growing in fibroin or GO plus fibroin films showed a discrete proliferation. Importantly, at day 10 of culture it was observed a significant increase in PDLSCs proliferation rate in GO films compared to plastic ( P  < 0.05), as well as in GO plus fibroin compared to fibroin alone ( P  < 0.001). Flow cytometry analysis showed that culture of PDLSCs in fibroin, GO or GO plus fibroin films did not significantly alter the level of expression of the mesenchymal markers CD73, CD90 or CD105 up to 168 h, being the cell viability in GO even better than obtained in plastic. Our findings suggest that the combination of human dental stem cells/fibroin/GO based-bioengineered constructs have strong potential for their therapeutic use in regenerative dentistry.