Health and environmental impacts of graphene-based materials need to be thoroughly evaluated before their potential applications. Graphene has strong cytotoxicity toward bacteria. To better understand its antimicrobial mechanism, we compared the antibacterial activity of four types of graphene-based materials (graphite (Gt), graphite oxide (GtO), graphene oxide (GO), and reduced graphene oxide (rGO)) toward a bacterial model-Escherichia coli. Under similar concentration and incubation conditions, GO dispersion shows the highest antibacterial activity, sequentially followed by rGO, Gt, and GtO. Scanning electron microscope (SEM) and dynamic light scattering analyses show that GO aggregates have the smallest average size among the four types of materials. SEM images display that the direct contacts with graphene nanosheets disrupt cell membrane. No superoxide anion (O(2)(•-)) induced reactive oxygen species (ROS) production is detected. However, the four types of materials can oxidize glutathione, which serves as redox state mediator in bacteria. Conductive rGO and Gt have higher oxidation capacities than insulating GO and GtO. Results suggest that antimicrobial actions are contributed by both membrane and oxidation stress. We propose that a three-step antimicrobial mechanism, previously used for carbon nanotubes, is applicable to graphene-based materials. It includes initial cell deposition on graphene-based materials, membrane stress caused by direct contact with sharp nanosheets, and the ensuing superoxide anion-independent oxidation. We envision that physicochemical properties of graphene-based materials, such as density of functional groups, size, and conductivity, can be precisely tailored to either reducing their health and environmental risks or increasing their application potentials.

Graphene oxide (GO) is a promising precursor to produce graphene-family nanomaterials for various applications. Their potential health and environmental impacts need a good understanding of their cellular interactions. Many factors may influence their biological interactions with cells, and the lateral dimension of GO sheets is one of the most relevant material properties. In this study, a model bacterium, Escherichia coli ( E. coli ), was used to evaluate the antibacterial activity of well-dispersed GO sheets, whose lateral size differs by more than 100 times. Our results show that the antibacterial activity of GO sheets toward E. coli cells is lateral size dependent. Larger GO sheets show stronger antibacterial activity than do smaller ones, and they have different time- and concentration-dependent antibacterial activities. Large GO sheets lead to most cell loss after 1 h incubation, and their concentration strongly influences antibacterial activity at relative low concentration (<10 μg/mL). In contrast, when incubating with small GO sheets up to 4 h, the inactivation rate of E. coli cells continues increasing. The increase of small GO sheet concentration also results in persistent increases in their antibacterial activity. In this study, GO sheets with different lateral sizes are all well dispersed, and their oxidation capacity toward glutathione is similar, consistent with X-ray photoelectron spectroscopy and ultraviolet-visible absorption spectroscopy results. This suggests the lateral size-dependent antibacterial activity of GO sheets is caused by neither their aggregation states, nor oxidation capacity. Atomic force microscope analysis of GO sheets and cells shows that GO sheets interact strongly with cells. Large GO sheets more easily cover cells, and cells cannot proliferate once fully covered, resulting in the cell viability loss observed in the followed colony counting test. In contrast, small GO sheets adhere to the bacterial surfaces, which cannot effectively isolate cells from environment. This study highlights the importance of tailoring the lateral dimension of GO sheets to optimize the application potential with minimal risks for environmental health and safety.

To further our understanding on the antibacterial activity of single-walled carbon nanotubes (SWCNTs), high purity SWCNTs with average diameter of 0.83 nm and (7,5) chirality as dominate (n,m) structure were dispersed in a biocompatible surfactant solution. Ultraviolet-visible-near-infrared radiation absorption spectroscopy was employed to monitor the aggregation of SWCNTs. The results demonstrated that individually dispersed SWCNTs were more toxic than SWCNT aggregates toward bacteria (gram-negative Escherichia coli, Pseudomonas aeruginosa, and gram-positive Staphylococcus aureus, Bacillus subtilis). Individually dispersed SWCNTs can be visualized as numerous moving "nano darts" in the solution, constantly attacking the bacteria; thereby, degrading the bacterial cell integrity and causing the cell death. Controlled experimental results suggested that inhibiting cell growth and oxidative stress were not the major causes responsible for the death of cells. Furthermore, the detrimental effects of Co metal residues (up to 1 mug/mL) on SWCNT samples can be ruled out. Atomic force microscope study conducted in suspension proved that the death rates of bacteria were strongly correlated with their mechanical properties; soft cells were more vulnerable to SWCNT piercing. The antibacterial activity of SWCNTs can be remarkably improved by enhancing the SWCNT physical puncture on bacteria in the following ways: (1) dispersing SWCNTs individually to sharpen the nano darts; (2) increasing SWCNT concentration to raise the population density of nano darts; and (3) elevating the shaking speed of incubation to speed up the nano darts. This study elucidated several factors controlling the antibacterial activity of pristine SWCNTs and it provided an insight in developing strategies that can maximize the SWCNT application potentials while minimizing the health and environment risks.

Single-walled carbon nanotubes (SWCNTs) exhibit strong antibacterial activities. Direct contact between bacterial cells and SWCNTs may likely induce cell damages. Therefore, the understanding of SWCNT-bacteria interactions is essential in order to develop novel SWCNT-based materials for their potential environmental, imaging, therapeutic, and military applications. In this preliminary study, we utilized atomic force microscopy (AFM) to monitor dynamic changes in cell morphology and mechanical properties of two typical bacterial models (gram-negative Escherichia coli and gram-positive Bacillus subtilis) upon incubation with SWCNTs. The results demonstrated that individually dispersed SWCNTs in solution develop nanotube networks on the cell surface, and then destroy the bacterial envelopes with leakage of the intracellular contents. The cell morphology changes observed on air dried samples are accompanied by an increase in cell surface roughness and a decrease in surface spring constant. To mimic the collision between SWCNTs and cells, a sharp AFM tip of 2 nm was chosen to introduce piercings on the cell surface. No clear physical damages were observed if the applied force was below 10 nN. Further analysis also indicates that a single collision between one nanotube and a bacterial cell is unlikely to introduce direct physical damage. Hence, the antibacterial activity of SWCNTs is the accumulation effect of large amount of nanotubes through interactions between SWCNT networks and bacterial cells.

Biochemical, structural, and functional properties of Rab5 wild-type (WT) protein were compared with those of Q79L and N133I mutants. The detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate increased guanine nucleotide binding to Rab5 WT approximately 10-fold. The single-step catalytic rate of Rab5 WT exceeded that of Q79L 12.2-fold, but the steady-state GTPase rate was only 2.8-fold greater because GDP dissociation was rate-limiting and GDP dissociation was 3.6-fold slower than for Q79L. In contrast, dissociation rates of GTP were indistinguishable. Binding to Rab5 N133I was not detectable. GTP protected Rab5 WT and Q79L from any apparent proteolysis by trypsin. A 20-kDa fragment was the major product of digestion in the presence of GDP, and 12- and 8-kDa fragments were the major products in the absence of added guanine nucleotides. Rab5 N133I underwent no apparent proteolysis with 10 mM GTP or GDP, suggesting a "triphosphate" conformation may be induced in Rab5 N133I by either GTP or GDP. Partially geranylgeranylated Rab5 WT stimulated endosome fusion in vitro, whereas unmodified Rab5 WT did not. Processed Rab5 Q79L failed to inhibit endosome fusion, and Rab5 N133I could not be geranylgeranylated. These findings identify biochemical and structural features of Rab5 proteins, providing data for the interpretation of functional assays.