Examples of Biomedical Research Using Graphene and Graphene Oxide

【A】Examples of Antibacterial Research

No. Platform Bacteria Type Result Literature
A-1 monolayer graphene film on conductor Cu, semiconductor Ge and insulator SiO2 S. aureus and E. coli graphene films on Cu and Ge can surprisingly inhibit the growth of both bacteria, especially the former; not significantly inhibited by the graphene film on SiO2 Li, J.; Wang, G.; Zhu, H.; Zhang, M.; Zheng, X.; Di, Z.; Liu, X.; Wang, X. Antibacterial activity of large-area monolayer graphene film manipulated by charge transfer. Nature Scientific Reports 2014, 4: 4359
A-2 graphene oxide (GO), reduced graphene oxide (rGO) nanosheets E. coli Both GO and rGO can effectively inhibit the growth of E. coli bacteria while showing minimal cytotoxicity. Hu, W. B.; Peng, C.; Luo, W. J.; Lv, M.; Li, X.; Li, D.; Huang, Q.; Fan, C. Graphene-based antibacterial paper. ACS Nano 2010, 4, 4317-4323
A-3 graphite (Gt), graphite oxide (GtO), graphene oxide (GO), reduced graphene oxide (rGO) E. coli Under similar concentration and incubation conditions, GO dispersion shows the highest antibacterial activity, sequentially followed by rGO, Gt, and GtO. Results suggest that antimicrobial actions are contributed by both membrane and oxidation stress. Liu, S.; Zeng, T.H.; Hofmann, M.; Burcombe, E.; Wie, E.; Jiang, R.; Kong, R. and Chen, R.　Antibacterial activity of graphite, graphite oxide, graphene oxide and reduced graphene oxide: Membrane and oxidative stress. ACS Nano 2011, 5, 6971-6980
A-4 graphene oxide (GO), reduced graphene oxide (rGO) P. aeruginosa GO and rGO showed dose-dependent antibacterial activity against P. aeruginosa cells through the generation of reactive oxygen species (ROS) , leading to cell death. Gurunathan, S.; Han, J.W.; Dayem, A.A.; Eppakayala, V. and Kim, H. Oxidative stress-mediated antibacterial activity of graphene oxide and reduced graphene oxide in Pseudomonas aeruginosa. Int. J. Nanomed. 2012, 7, 5901-5914.
A-5 graphene, graphene oxide (GO), reduced graphene oxide (rGO) nanowalls S. aureus and E. coli reduced graphene oxide (rGO) showed higher toxicity for S. aureus than GO and graphene, but E. coli is more resistant due to the presence of outer membrane. Akhavan, O. and Ghaderi, E. Toxicity of graphene and graphene oxide nanowalls against bacteria. ACS Nano., 2010, 4:5731-5736.

【B】Examples of Antiviral Research

No. Platform Virus type Result Literaturte
B-1 graphene oxide (GO)、reduced graphene oxide (rGO) pseudorabies virus (PRV), porcine epidemic diarrhea virus (PEDV) GO significantly suppressed PRV and PEDV infection and was shown to exhibit broad spectrum antiviral activity at non-cytotoxic concentrations (6 μg/mL). The antiviral activity is time and concentration dependent. Ye, S.; Shao, K.; Li, Z.; Guo, N.; Zuo, Y.; Li, Q.; Lu, Z.; Chen, L.; He, Q. and Han, H. Antiviral Activity of Graphene Oxide: How Sharp Edged Structure and Charge Matter. ACS Appl. Mater. Interfaces 2015, 7, 38, 21571-21579
B-2 garaphene oxide (GO), graphene oxide-Ag nanocomposites (GO-Ag) feline coronavirus (FCoV), infectious bursal disease virus (IBDV) Go-Ag inhibited 25% of infection by FCoV and 23% by IBDV, whereas GO only inhibited 16% of infection by FCoV but showed no antiviral activity against the infection by IBDV. Chen, Y.N.; Hsueh, Y.H.; Hsieh, C.T.; Tzou, D.Y.; Chang, P.L. Antiviral Activity of Graphene Silver
Nanocomposites against Non-Enveloped and Enveloped Viruses. Int. J. Environ. Res. Public Health 2016, 13(4), 430
B-3 seven different carbon quantum dots (CQDs) human coronavirus (HCoV-229E) Three of the seven CQDs (CQD-3, -5, -6) have been shown to significantly interfere with HCoV-229E-Luc infection in a concentration-dependent manner. A. oczechin, K. Seron, A. Barras, E. Giovanelli, S. Belouzard, Y. T. Chen, N. Metzler-Nolte, R. Boukherroub, J. Dubuisson and S. Szunerits, Functional Carbon Quantum Dots as Medical Countermeasures to Human Coronavirus. ACS Appl. Mater. Interfaces, 2019, 11, 42964-42974.
B-4 sulfonated magnetic nanoparticles functionalized with reduced graphene oxide (SMRGO) herpes simplex virus type 1（HSV-1） SMRGO showed effective and rapid (~ 99.99%, 7 minutes) antiviral activity. Deokar, A.R.; Nagvenkar, A.P.; Kalt, I; Shani, L.; Yeshurun, Y.; Gedanken, A.; Sarid, R. Graphene-Based “Hot Plate” for the Capture and Destruction of the Herpes Simplex Virus Type 1. Bioconjugate Chem. 2017, 28, 1115-1122
B-5 2,2′-(ethylenedioxy)bis(ethylamine) (EDA)-CDots and 3-ethoxypropylamine (EPA)-CDots, human norovirus virus-like-particles (GI.1 and GII.4 VLPs) both EDA- and EPA- CDots were highly effective to inhibit both strains of VLPs’ bindings to histo-blood group antigens (HBGA) receptors on human cells at CDots concentration of
5μg/mL, with EDA-CDots achieving 100% inhibition and EPA CDots achieving 85-99% inhibition.		 Dong, X.; Moyer, M.M.; Yang, F.; Sun, Y.P.; Yang, L. Carbon Dots’ Antiviral Functions Against Noroviruses. Nature Scientific Reports 2017, 7: 519

【C】Examples of Research on Biosensors

No. Sensing element Sensor type Sensor platform Limit of detection（μM） Linear range（mM） References
C-1 glucose electrochemical CuO-graphene 1 0.001～8 Hsu, Y.-W.; Hsu, T.-K.; Sun, C.-L.; Nien, Y.-T.; Pu, N.-W.; Ger, M.-D. Synthesis of CuO/graphene nanocomposites for nonenzymatic electrochemical glucose biosensor applications. Electrochim. Acta 2012, 82, 152-157.
C-2 NADH photoelectrochemical graphene-TiO2 nanohybrids 0.003 0.00001～2 Wang, K.; Wu, J.; Liu, Q.; Jin, Y.; Yan, J.; Cai, J. Ultrasensitive photoelectrochemical sensing of nicotinamide adenine dinucleotide based on graphene-TiO2 nanohybrids under visible irradiation. Anal. Chim. Acta 2012, 745, 131-136.
C-3 DNA electrochemical polyaniline/graphene 1 × 10-8 1×10-10～1×10-3 Zheng, Q.; Wu, H.; Shen, Z.; Gao, W.; Yu, Y.; Ma, Y.; Guang, W.; Guo, Q.; Yan, R.; Wang, J.; Ding, K. An electrochemical DNA sensor based on polyaniline/graphene: high sensitivity to DNA sequences in a wide range. Analyst 2015, 140 (19), 6660-6670.
C-4 H2O2 electrochemical horseradish peroxidase-MoS2-graphene 0.049 0.0002～1.103 Song, H.; Ni, Y.; Kokot, S. Investigations of an electrochemical platform based on the layered MoS2 graphene and horseradish peroxidase nanocomposite for direct electrochemistry and electrocatalysis. Biosens. Bioelectron. 2014, 56, 137-143.
C-5 cancer cells electrochemical graphene-peptide nanotube-folic acid 250 cells/mL 　 Castillo, J. J.; Svendsen, W. E.; Rozlosnik, N.; Escobar, P.; Martinez, F.; Castillo-Leon, J. Detection of cancer cells using a peptide nanotube folic acid modified graphene electrode. Analyst 2013, 138 (4), 1026-1031.
C-6 immunoglobulin-G flourescence graphene- mouse antihuman immunoglobulin G (mIgG)-graphene quantum dots (GQDs) 10 ng/mL 0.2～12 μg/mL Zhao, H.; Chang, Y.; Liu, M.; Gao, S.; Yu, H.; Quan, X. A universal immunosensing strategy based on regulation of the interaction between graphene and graphene quantum dots. Chem. Commun. 2013, 49 (3), 234-236.
C-7 carcinoembryonic
antigen		 FET (Field Effect Transistor) graphene-anti-CEA ( carcinoembryonic antigen) 100 pg/mL 　 Zhou, L.; Mao, H.; Wu, C.; Tang, L.; Wu, Z.; Sun, H.; Zhang, H.; Zhou, H.; Jia, C.; Jin, Q.; et al. Label-free graphene biosensor targeting cancer molecules based on non-covalent modification. Biosens. Bioelectron. 2017, 87, 701-707.

【D】Examples of Research on Tissue Engineering

No. Cell Type Commitment toward tissue engeneering/tissue regeneration Results/Uses References
D-1 mesenchymal stem cells (MSCs) cardiac graphene promotes cardiomyogenic differentiation without any cytotoxicity Park, J.; Park, S.; Ryu, S.; Bhang, S. H.; Kim, J.; Yoon, J. K.;　Park, Y. H.; Cho, S. P.; Lee, S.; Hong, B. H.; Kim, B.-S. GrapheneRegulated Cardiomyogenic Differentiation Process of Mesenchymal Stem Cells by Enhancing the Expression of Extracellular Matrix Proteins and Cell Signaling Molecules. Adv. Healthcare Mater. 2014, 3(2), 176-181.
D-2 human neural stem cells (HNSCs) neuronal enhanced stem cells differentiation into neurons Park, S. Y.; Park, J.; Sim, S. H.; Sung, M. G.; Kim, K. S.; Hong, B. H.; Hong, S. Enhanced differentiation of human neural stem cells into neurons on graphene. Adv. Mater. 2011, 23 (36), H263.
D-3 mesenchymal stem cells (MSCs), osteoblast
cells		 bone tissue graphene induces osteoblast cell proliferation, on graphene coated substrates cells are adhered and proliferated better than SiO2 substrate Kalbacova, M.; Broz, A.; Kong, J.; Kalbac, M. Graphene substrates promote adherence of human osteoblasts and mesenchymal stromal cells. Carbon 2010, 48 (15), 4323-4329.
D-4 human neural stem cells (MSCs) skin tissue graphene foam guided the wound healing process in a faster way with reduced scarring effect Li, Z.; Wang, H.; Yang, B.; Sun, Y.; Huo, R. Three dimensional graphene foams loaded with bone marrow derived mesenchymal stem cells promote skin wound healing with reduced scarring. Mater. Sci. Eng., C 2015, 57, 181-188.

【E】Examples of Research on Drug/Gene Delivery System)

No. Carrier Delivered drugs/genes Results/Uses References
E-1 graphene-SiO2 quoted quantum dots
(HQDs) with Transferrin		 doxorubicin (DOX) hybrid system efficiently delivers DOX to the targeted cancer cells; enables to monitor the intracellular DOX release Chen, M.-L.; He, Y.-J.; Chen, X.-W.; Wang, J.-H. Quantum dot-conjugated graphene as a probe for simultaneous cancer-targeted
fluorescent imaging, tracking, and monitoring drug delivery. Bioconjugate Chem. 2013, 24 (3), 387-397.
E-2 hyaluronic acid-functionalized graphene quantum dot conjugated
albumin nanoparticles		 gemcitabine (GMC) targeted delivery of GMC was observed with sustained release behavior, very good toxicity was showed by GMC loaded nanocarrier system toward pancreatic cancer cells Nigam, P.; Waghmode, S.; Louis, M.; Wangnoo, S.; Chavan, P.; Sarkar, D. Graphene quantum dots conjugated albumin nanoparticles for targeted drug delivery and imaging of pancreatic cancer. J. Mater. Chem. B 2014, 2 (21), 3190-3195.
E-3 PEGylated graphene/Au composites siRNA gene silencing (Bcl-2) Cheng, F.-F.; Chen, W.; Hu, L.-H.; Chen, G.; Miao, H.-T.; Li, C.; Zhu, J.-J. Highly dispersible PEGylated graphene/Au composites
as gene delivery vector and potential cancer therapeutic agent. J. Mater. Chem. B 2013, 1 (38), 4956-4962.
E-4 polyethylenimine-functionalized graphene oxide pDNA expression of endogenous genes EGFP Chen, B.; Liu, M.; Zhang, L.; Huang, J.; Yao, J.; Zhang, Z. Polyethylenimine-functionalized graphene oxide as an efficient gene delivery vector. J. Mater. Chem. 2011, 21 (21), 7736-7741.
E-5 folic acid conjugated nano graphene oxide doxorubicin (DOX), camptothecin (CPT) targeted delivery of DOX and CPT to MCF-7 cells with enhanced toxicity than single DOX or CPT Zhang, L.; Xia, J.; Zhao, Q.; Liu, L.; Zhang, Z. Functional graphene oxide as a nanocarrier for controlled loading and targeted delivery of mixed anticancer drugs. Small 2010, 6 (4), 537-544.

【F】Examples of Research on Bioimaging

No. Platform Imaging Tool Use References
F-1 graphene quantum dots(GQD) Flourescence Imaging of neurospheres stem cells  Zhang, M.; Bai, L.; Shang, W.; Xie, W.; Ma, H.; Fu, Y.; Fang, D.; Sun, H.; Fan, L.; Han, M.; et al. Facile synthesis of water-soluble,
highly fluorescent graphene quantum dots as a robust biological label for stem cells. J. Mater. Chem. 2012, 22 (15), 7461−7467.
F-2 graphene quantum dots(GQD) Flourescence imaging of Hela229 cells  Pan, D.; Guo, L.; Zhang, J.; Xi, C.; Xue, Q.; Huang, H.; Li, J.; Zhang, Z.; Yu, W.; Chen, Z.; et al. Cutting sp 2 clusters in graphene
sheets into colloidal graphene quantum dots with strong green fluorescence. J. Mater. Chem. 2012, 22 (8), 3314−3318.
F-3 microwave enabled low oxygen graphene nanosheets Photoacoustic fabricated graphene nanosheets exhibited strong NIR absorption and high photoacoustic conversion efficiencies, which suggests its
applicability for deep tissue imaging		  Patel, M. A.; Yang, H.; Chiu, P. L.; Mastrogiovanni, D. D.; Flach, C. R.; Savaram, K.; Gomez, L.; Hemnarine, A.; Mendelsohn, R.; Garfunkel, E.; et al. Direct production of graphene nanosheets for near infrared photoacoustic imaging. ACS Nano 2013, 7 (9), 8147−8157.
F-4 graphene oxide decorated with gold nanoparticles Raman imaging of Hela cells  Liu, Q.; Wei, L.; Wang, J.; Peng, F.; Luo, D.; Cui, R.; Niu, Y.; Qin, X.; Liu, Y.; Sun, H.; et al. Cell imaging by graphene oxide based
on surface enhanced Raman scattering. Nanoscale 2012, 4 (22), 7084−7089.
F-5 polyethylene glycol functionalized nano graphene oxide
conjugated with Rituxan		 NIR flourescence CD20 positive Raji B-cells Sun, X.; Liu, Z.; Welsher, K.; Robinson, J. T.; Goodwin, A.; Zaric, S.; Dai, H. Nano-graphene oxide for cellular imaging and drug
delivery. Nano Res. 2008, 1 (3), 203−212.