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

Abstract

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.

Cell survival and differentiation with nanocrystalline glass-like carbon using substantia nigra dopaminergic cells derived from transgenic mouse embryos

“…This replicative and proliferative processes, suppose a greater energetic cost for the cell and coincides with an increase of cellular metabolism and Calcein AM activity. Although time responses have been carried out in graphene studies [42] it was used heterogeneous cell cultures, where cells are metabolically more active than others, and this fact may be hiding the true action on MTT. Since SN4741 cells are clonogenic, we performed experiments to synchronize the cell growth of SN4741 cells prior to start the present study [43], so that it allows us to determine changes in cell cycle.…”

Combined Application of Graphene‐Family Materials and Silk Fibroin in Biomedicine

“…RodrÍguez‐Lozano et al. fabricated composite membranes of GO and SF via water annealing . Compared with pure SF membranes, the composite membranes increased the adhesion rate and promoted proliferation of periodontal ligament stem cells (PDLSCs).…”

“…Since graphene‐family materials were introduced into dental field, the combined use of graphene‐family materials and SF will provide a more effective strategy . The composite membranes of GO and SF fabricated by RodrÍguez‐Lozano have good biocompatibility and can significantly promote cell proliferation of PDLSCs . Vera‐Sánchez et al.…”

Graphene Oxide/Polymer‐Based Biomaterials

“…Rodriguez-Lozano et al [112] studied the effect of GO/ SF nanocomposite films on periodontal ligament stem cells. The morphology of periodontal ligament stem cells was examined for 10 days after staining with DAPI and phalloidin.…”