Treatment for Myocardial Infarction: In Vivo Evaluation of Curcumin-Loaded PEGylated-GQD Nanoparticles.

ABSTRACT: Curcumin (Cur) has been suggested as a complementary treatment for cardiovascular diseases. Its efficiency, however, is modest due to poor biocompatibility. This study examined the effects of curcumin loaded on polyethylene glycol-graphene quantum dots (Cur-PEG-GQDs) on hemodynamic and cardiac function in rats with myocardial infarction (MI). The study groups included control, MI, MI+Cur-3, MI + Cur-7, MI + Cur-15, MI + PEG-GQDs-5, MI + PEG-GQDs-10, MI + Cur-PEG-GQDs-5, and MI + Cur-PEG-GQDs-10. MI was established by left anterior descending artery ligation. Two weeks after intraperitoneal administration of vehicle, Cur, PEG-GQDs, and Cur-PEG-GQDs, blood pressure and heart contractility indices were measured. Triphenyl tetrazolium chloride, colorimetry, and clinical laboratory methods were used to measure the infarct size, the oxidant and antioxidant content, and the kidney and liver function parameters, respectively. In the MI animals, Cur-7, PEG-GQDs-10, Cur-PEG-GQDs-5, and Cur-PEG-GQDs-10 recovered systolic blood pressure, diastolic blood pressure, left ventricular systolic pressure, and ±dp/dt max disturbances and reduced myocardial infarct size, fibrosis, and left ventricular end-diastolic pressure. Curcumin lowered antioxidant markers and elevated 1 oxidant marker in the heart in a dose-dependent manner. Although Cur-PEG-GQDs-5 and Cur-PEG-GQDs-10 reduced curcumin's oxidative stress effects, the superoxide dismutase, glutathione peroxidase, and total antioxidant capacity levels were significantly lower in Cur-PEG-GQDs-5 and Cur-PEG-GQDs-10 groups compared with the MI group. Malondialdehyde levels were lower in Cur-PEG-GQDs-5 and -10 groups compared with the Cur-3, Cur-7, and Cur-15 groups. The glutathione/glutathione disulfide ratio improved in the groups treated by Cur-7, PEG-GQDs-10, Cur-PEG-GQDs-5, and Cur-PEG-GQDs-10. The findings indicated that Cur-PEG-GQDs mitigated MI-induced cardiac dysfunction. However, because of the increase in oxidative stress in the heart, nonclassic mechanisms may be involved in the beneficial effect of Cur-PEG-GQDs on MI-induced cardiac dysfunction.

Beneficial effects of PEGylated graphene quantum dot on arrhythmias induced by myocardial infarction.

Arrhythmias are one of the leading causes of early death following myocardial infarction (MI) and heart failure. Graphene derivatives have emerged as an therapeutic target that have electrical conductivity. The study aimed to evaluate the impacts of polyethylene glycol-graphene quantum dots (GQDs-PEG) on arrhythmias created by MI in the rat. Animals were randomly assigned to five groups of sham, MI, and MI + GQDs-PEG at doses of 5, 10, and 20 mg/kg. MI was induced by the closure of the left anterior descending artery. The day after MI, animals were administered vehicle (phosphate buffered saline) or GQDs-PEG at different doses every other day for 2 weeks. On day 15, electrocardiogram (ECG), mean arterial pressure (MAP), and heart contractility indices were recorded by the PowerLab data acquisition system. GQDs-PEG 20 mg/kg increased contractility and improved the reduction of MAP in the MI group. The prolonged QT and QTc intervals, inverted T wave, and deviated ST segment were modified by GQDs-PEG 10 and 20 mg/kg in rats with MI. The amplitude of the Q wave was also decreased in a dose-dependent manner in the GQDs-PEG-treated rats. The results demonstrated that 2 weeks of treatment with GQDs-PEG normalized ECG abnormalities and improved left ventricular dysfunction in rats with MI.

Preparation, Characterization, and Wound Healing Assessment of Curcumin-Loaded M-MOF (M = Cu, Zn)@Polycaprolactone Nanocomposite Sponges.

The fabrication of multifunctional scaffolds has attracted much attention in biological fields. In this research, some novel composites of Cu(II) or Zn(II) metal-organic framework (M-MOF) and polycaprolactone (PCL), M-MOF@PCL, have been fabricated as multifunctional scaffolds for application in the tissue engineering (TE) field. The porous three-dimensional sponges were prepared by the salt leaching method. Then, the M-MOF@PCL composite sponges have been prepared by in situ synthesis of M-MOF in the presence of the as-obtained PCL sponge to gain a new compound with proper features for biological applications. Finally, curcumin was attached to the M-MOF@PCL as a bioactive compound that can act as a wound-healing agent, anti-oxidant, and anti-inflammatory. The presence of the M-MOF in final composites was investigated by different methods such as FTIR (Fourier-transform infrared), XRD (X-ray diffraction), SEM (scanning electron microscope), EDS (energy-dispersive X-ray spectroscopy), and TEM (transmission electron microscope). SEM images confirmed the porous structure of the as-obtained composites. According to the EDS and TEM images, M-MOFs were uniformly incorporated throughout the PCL sponges. The water sorption capacities of the blank PCL, Cu-MOF@PCL, and Zn-MOF@PCL were determined as 56%, 155%, and 119%, respectively. In vivo investigation on a third-degree burn model in adult male Wistar rats exhibited an accelerated wound healing for Cu-MOF@PCL compared to with Zn-MOF@PCL and the control group.

PEGylated Graphene Quantum Dot Improved Cardiac Function in Rats with Myocardial Infarction: Morphological, Oxidative Stress, and Toxicological Evidences.

INTRODUCTION: The biocompatibility and potential application of graphene-based nanomaterials in biomedicine have been documented. The effects of polyethylene glycol-graphene quantum dots (GQDs-PEG) on cardiac function in rats with myocardial infarction (MI) were examined. METHODS: Wistar rats were randomly assigned to two main groups, each consisting of sham-Veh., MI-Veh., and MI+GQDs-PEG at doses of 5, 10, and 20 mg/kg. MI was induced by the closure of the left anterior descending (LAD) coronary artery. After MI, GQDs-PEG were injected at different doses IP every other day for two weeks. In the end, hemodynamic and heart contractility indices were assessed. The levels of myocardial MDA (malondialdehyde), SOD (superoxide dismutase), GPX (glutathione peroxidase), and TAC (total antioxidant capacity) were measured by the ELISA method. The serum ALP, ALT, AST, creatinine, and urea levels were measured using the photometric method. The infarct size was assessed by TTC staining. RESULTS: GQDs-PEG decreased the infarct size at doses of 10 and 20 mg/kg and recovered the MI-induced reductions of +dp/dt max and -dp/dt max in the study groups. GQDs-PEG normalized systolic blood pressure and left ventricular systolic pressure reduction at the dose of 20 mg/kg in the MI group. Heart SOD, GPX, and TAC increased in the GQDs-PEG 10 and 20 groups. Almost no signs of toxic effects due to GQDs-PEG administration were observed on the liver and kidneys. CONCLUSIONS: The results provided clear evidence that GQDs-PEG improve cardiac performance and hemodynamic parameters in rats with MI by reducing oxidative stress. GQDs-PEG is proposed as a therapeutic target for the treatment of MI.

Curcumin loaded on graphene nanosheets induced cell death in mammospheres from MCF-7 and primary breast tumor cells.

Elimination of tumor cells is still a therapeutic challenge for breast cancer (BC) in men and women. Mammospheres serve as valuablein vitrotools for evaluating tumor behavior and sensitivity to anticancer treatments. Graphene nanosheets with unique physicochemical properties have been considered as potential biomedical approaches for drug delivery, bioimaging, and therapy. Graphene oxide (GO) and graphene quantum dots (GQDs) are suitable nanocarriers for hydrophobic and low bioaccessible anti-tumor materials like curcumin. Despite extensive studies on the potential application of graphene nanosheets in medicine, our knowledge of how different cells function and respond to these nanoparticles remains limited. Here, we evaluated cell death in mammospheres from MCF-7 and primary tumor cells in response to curcumin loaded on graphene nanosheets. Mammospheres were exposed to graphene oxide-curcumin (GO-Cur) and graphene quantum dots-curcumin (GQDs-Cur), and the incidence of cell death was evaluated by Hoechst 33342/propidium iodide double staining and flow cytometry. Besides, the expression of miR-21, miR-29a, Bax, and Bcl-2 genes were assessed using RT-qPCR. We observed, GO, and GQDs had no cytotoxic effect on Kerman male breast cancer/71 (KMBC/71) and MCF-7 tumor cells, while curcumin induced death in more than 50% of tumor cells. GO-Cur and GQDs-Cur synergistically enhanced anti-tumor activity of curcumin. Moreover, GQDs-Cur induced cell death in almost all cells of KMBC/71 mammospheres (99%;p< 0.0001). In contrast, GO-Cur induced cell death in only 21% of MCF-7 mammosphere cells (p< 0.0001). Also, the expression pattern of miR-21, miR-29a, and Bax/Bcl-2 ratio in KMBC/71 and MCF-7 mammospheres was different in response to GO-Cur and GQDs-Cur. Although KMBC/71 and MCF-7 tumor cells had similar clinical features and displayed similar responses to curcumin, more investigations are needed to clarify the detailed molecular mechanisms underlying observed differences in response to GO-Cur and GQDs-Cur.

Secondary toxic effect of graphene oxide and graphene quantum dots alters the expression of miR-21 and miR-29a in human cell lines.

For in vitro studies, non-toxic doses of nanomaterials are routinely selected by quantification of live cells after exposing to different concentrations of nanoparticles but considering only morphological changes or viability of cells is not sufficient to conclude that these nanomaterials are non-cytotoxic. Here we investigated if secondary toxicity is active in the cells exposed to non-toxic doses of graphene oxide (GO) and graphene quantum dots (GQDs). Non-cytotoxic dose of 15 µg mL-1 of GO (100 nm) and GQDs (50 nm) was selected according to MTT and Hoechst 33342/PI double staining assays. In order to investigate the secondary toxicity, the expression of miR-21, miR-29a and three genes at both mRNA and protein level were evaluated in MCF-7, HUVEC, KMBC/71 cells 4 and 24 h post exposure. Mitochondrial membrane potential (MMP) was assessed by Rhodamine 123 staining. According to our results, there was no significant decrease in viability of cells after exposure to the non-cytotoxic dose of GO and GQDs, but we observed significant alterations in the expression level of miR-21, miR-29a, Bax, Bcl2 and PTEN genes after treatment in all three cells. In addition to molecular changes, we observed alteration in mitochondrial activity at cellular level. However, we also observed that GO influenced the basal level of genes and MMP more compare to GQDs. Considering that all these genes are involved in breast tumor development and metastasis, the observed changes in miRNA expression and protein synthesis may alter cell fate and susceptibility and cause deviation in the desired outcome of GO and GQDs application in medical research.

Fabrication and biocompatibility assessment of polypyrrole/cobalt(II) metal-organic frameworks nanocomposites.

Nowadays, metal-organic frameworks (MOFs) have emerged as promising tools for different biological applications and therefore, efforts are ongoing to develop more biocompatible MOFs-based nanocomposites. We aimed to fabricate some new conductive nanocomposites of polypyrrole and cobalt-MOF with different weight percentages (PPy/x%Co-MOF) using the solution mixing method and characterize them through FT-IR (Fourier-transform infrared), PXRD (powder X-ray diffraction), SEM (scanning electron microscope), and TEM (transmission electron microscope) techniques. The biocompatibility of nanocomposites was assessed by haemolytic, cytotoxic, and quantitative reverse transcription PCR (qRT-PCR) assays. FT-IR and PXRD results revealed that nanocomposites consisted of pure MOFs and PPy. Moreover, SEM results indicated their spherical morphology along with an average diameter of 190 nm. (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed a concentration, and percentagedependent cytotoxic effect of the nanocomposites on some cell lines including 3T3 fibroblasts, MCF-7, and J774.A1 macrophages. Haematological toxicity of PPy/x%Co-MOF composites was less than 7% in most concentrations. Furthermore, PPy/x%Co-MOF composites did not show any significant effect on the expression of cyclooxygenase-2( COX-2) and inducible nitric oxide synthase( iNOS) genes. In sum, regarding the haemolytic, proinflammatory, and cytotoxic tests, prepared nanocomposite demonstrated the reasonable in vitro biocompatibility which may be considered as a hopeful platform for further investigations including clinical applications.

Synthesis, characterization and biocompatibility of polypyrrole/Cu(II) metal-organic framework nanocomposites.

The main objective of composite science is to fabricate new materials with desired properties such as high chemical, mechanical, and/or biological performances. In this research, new conductive nanocomposites of copper metal-organic frameworks (Cu-MOF) and polypyrrole (PPy) were fabricated with the aim of exploiting the electrical conductivity of polypyrrole and the porosity of MOFs in the final products. The prepared compounds (PPy/x%Cu-MOF, x = 20, 50, and 80) were investigated by FTIR, PXRD, SEM, TEM, DLS, BET, EDS mapping, cyclic voltammetry (CV), and zeta potential (ξ) measurements. Spherical morphology was confirmed by SEM and TEM analysis. The PPy/80%Cu-MOF nanocomposite showed the highest ξ potential (-40 mV), demonstrating the stability of dispersed particles. The CV results revealed that the nanocomposites have higher capacitance in comparison to the pure materials. In vitro degradation of the as-prepared compounds in simulated body fluid (SBF) was studied by EIS (electrochemical impedance spectroscopy) and Tafel polarization tests. Furthermore, in vitro biocompatibility of the PPy/x%Cu-MOF composite was evaluated on a group of cells including 3T3 fibroblasts, MCF-7 breast cancer cells, J774.A1 macrophages and red blood cells (RBCs). Viability of 3T3 fibroblasts, MCF-7, and J774.A1 cells, by Methylthiazolyldiphenyl-tetrazolium bromide (MTT) method, was dependent on Cu-MOF percent and amount of composites. Hemolytic assay for RBCs exposed to different amounts of the PPy/x%Cu-MOF composites showed hematological toxicity less than 5% in most concentrations. In addition, to investigate pro-inflammatory activity, J774.A1 macrophages were exposed to non-toxic concentrations of the PPy/x%Cu-MOF and no significant change in the expression of two inflammatory genes COX-2 and iNOS was observed. Injection of the PPy/x%Cu-MOF (5 mg kg-1) into bloodstream of mice did not increase liver damage marker enzymes alanine transaminase (ALT) and aspartate transaminase (AST) level in serum 1 week post injection. Moreover, we observed slight but not significant increase in serum copper level in mice 1 week after injection. According to the results, the PPy/x%Cu-MOF nanocomposites exhibited a good in vitro and in vivo biocompatibility without inducing pro-inflammatory responses in macrophages and show promising potential for different biomedical applications such as biosensors and drug delivery. The release of curcumin from curcumin-loaded PPy/x%Cu-MOF nanocomposites was detectable in plasma of mice 4 days after administration.