Few layer graphene does not affect the function and the autophagic activity of primary lymphocytes

Abstract

Primary lymphocytes ensure their efficient function and maintenance through different mechanisms including autophagy. Few layer graphene does not affect either the viability and activation or the autophagic activity of B and T cells.

Few Layer Graphene Does Not Affect Cellular Homeostasis of Mouse Macrophages

“…To investigate how FLG influences autophagy in primary macrophages, BMDMs were incubated with low (10 µg/mL) and high (100 µg/mL) doses of graphene for 24 h. A representative immunoblot (Figure 2A) of cell lysates of untreated, low-and high-dose FLG-treated and LPS-treated conditions shows the expression of LC3-I and -II with or without lysosomal protease inhibitors (E64D and pepstatin A). The expression of LC3 proteins in low-and high-dose FLG-treated cells is similar to basal level as observed in untreated cells and LPS-treated cells [22] and was observed to be slightly higher in the presence of protease inhibitors, indicating the inhibition of the degradation of autophagosomes. The visualization of LC3 bands is, however, not enough to attest to the influence of FLG on autophagic activity induction in macrophages, hence the densitometric ratio was calculated between membrane-bound LC3-II protein and β-actin (internal loading reference).…”

“…Recently, it has been reported that FLG can induce oxidative stress and endoplasmic-mediated autophagy in RAW264.7 macrophages [21]. In primary T and B cells, in contrast, our group has found that FLG does not impact viability and activation, nor autophagic activity [22].…”

“…The final FLG water dispersions were lyophilized and the final graphene powder was characterized and used in the cell experiments following the dispersion in culture media [19]. The characterization of FLG by elemental analysis, TEM and Raman spectroscopy, was previously published by Murera et al [22]. Briefly, the lateral size of the nanomaterial ranges from 100 nm to 1600 nm, the Raman average spectrum of FLG is composed of the typical graphene Raman bands: G peak, D peak, and 2D band, which are centered at~1580 cm −1 ,~1350 cm −1 , and~2680 cm −1 , respectively, and elemental analysis showed that the sample is mainly composed of carbon, with a maximum of 0.93 ppm of melamine traces.…”

Toxicological evaluation of highly water dispersible few-layer graphene in vivo

“…Recently, a comprehensive study has been published demonstrating how FLG does not affect the function and the autophagic activity of primary lymphocytes. [30] The results showed that FLG neither impacts the viability and activation of T and B cells nor their autophagic activity. Before studying the effects of G-Rib in an animal model, we wanted to evaluate the cytotoxic effects in two different cell lines.…”

Modulation of Cancer Cell Autophagic Responses by Graphene-Based Nanomaterials: Molecular Mechanisms and Therapeutic Implications

“…While in some studies cytotoxicity was observed [95][96][97][98][99], GNM-induced autophagy in healthy, non-transformed primary cells was usually not associated with significant cell death [100][101][102][103][104][105][106]. The certain types of normal cells such as mouse lymphocytes and monkey fibroblasts were apparently resistant to both autophagy-modulating and cytotoxic action of GNM [107,108]. Interestingly, in a direct comparison under the same experimental conditions, GO/chloroquine conjugate at moderate concentration (25 µg/mL) induced autophagy-dependent necroptosis in lung carcinoma cells, but not in their non-cancerous counterparts [43].…”