Synergistic effects of Metformin and Forskolin on oxidative stress induced by diabetes and hepatocellular cancer: An animal study.

AIM: This study proposed to assess the synergistic effects of Forskolin and Metformin (alone and in combination) on glucose, hematological, liver serum, and oxidative stress parameters in diabetic, healthy, and hepatocellular carcinoma (HCC) induced rats. MATERIALS AND METHODS: Eighty male Wistar rats were divided into 10 experimental groups (8 rats for each group), including 1) healthy group, 2) diabetic group, 3) HCC group, 4) diabet + Metformin (300 mg/kg), 5) diabet + Forskolin (100 mg/kg), 6) diabet + Metformin (300 mg/kg) & Forskolin (100 mg/kg), 7) HCC + Metformin (300 mg/kg), 8) HCC + Forskolin (100 mg/kg), 9) HCC + Metformin (300 mg/kg) & Forskolin (100 mg/kg), and 10) healthy group + Metformin (300 mg/kg) & Forskolin (100 mg/kg). The rats were administrated Forskolin/Metformin daily for 8 weeks. Glucose, hematological, and liver serum parameters were measured and compared among the groups. The levels of malondialdehyde (MDA), and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx), as well as 8-hydroxydeoxyguanosine (8 OHdG) levels, were also measured. RESULTS: The average blood glucose reduction in diabetic rats with the Forskolin, Metformin, and Forskolin + Metformin treatments was 43.5%, 47.1%, and 53.9%, respectively. These reduction values for HCC rats after the treatments were 21.0%, 16.2%, and 23.7%, respectively. For all the diabetic and HCC rats treated with Forskolin/Metformin, the MDA, SOD, and GPx levels showed significant improvement compared with the diabetic and HCC groups (P < 0.05). Although the rats treated with Forskolin + Metformin experienced a higher reduction in oxidative stress of blood and urine samples compared to the Forskolin group, the differences between this group and rats treated with Metformin were not significant for all parameters. CONCLUSION: Metformin and Forskolin reduced oxidative stress in diabetic and HCC-induced rats. The results indicated that the combination of agents (Metformin & Forskolin) had greater therapeutic effects than Forskolin alone in reducing glucose levels in diabetic rats. However, the ameliorative effects of combining Metformin and Forskolin on blood and urine oxidative stress were not statistically higher than those of Metformin alone.

Forskolin Improves Male Reproductive Complications Caused by Hyperglycemia in Type 2 Diabetic Rats.

BACKGROUND: In many diabetic patients, spermatogenesis complications are frequent causing infertility problems. This study aimed to demonstrate the effect of Forskolin on male reproductive dysfunction caused by type 2 diabetes. MATERIALS AND METHODS: In this experimental study, type 2 diabetes was induced by a high-fat diet (HFD) for one month and then a low single dose injection (35 mg/kg) of streptozotocin (STZ) in Wistar rats. After 72 hours, rats with more than 200 mg/dl of blood glucose were considered type 2 diabetic rats. Forty rats (200-250 g) were divided into four groups (n=10) including group 1 (G1): rats with normal diet and buffer citrate (STZ solvent) injection, group 2 (G2): control type 2 diabetic rats with HFD and STZ injection, group 3 (G3): type 2 diabetic rats received phosphate buffer saline (PBS) as Forskolin solvent, and group 4 (G4): Forskolin treated diabetic rats (10 mg/kg) for 1 month. RESULTS: In comparison to control group, in diabetic groups (G2 and G3) some parameters are increased significantly: The blood glucose (P=0.00078), testicular malondialdehyde (MDA) level and body weight (P=0.00009) and Bax gene expression (P=0.00007). Unlike, some parameters are decreased significantly: The serum level of testosterone (P=0.0009), testicular superoxide dismutase (SOD, P=0.00007) and glutathione peroxidase (GPX) levels (P=0.00008), sperm concentration (P=0.00008), motility (P=0.00009), normal morphological sperm (P=0.00008) and Bcl-2 gene expression (P=0.00009). However, in Forskolin treated group (G4) the parameters stayed close to control values that was significantly (P=0.00007) higher than in G2 and G3 groups. Therefore, treatment with Forskolin significantly improved these abnormal changes in Forskolin-treated group. CONCLUSION: Our study demonstrates that Forskolin is an effective antidiabetic agent, which significantly improves sperm concentration, testosterone levels, and antioxidant activity in diabetic rats.

The effects of metformin and forskolin on sperm quality parameters and sexual hormones in type II diabetic male rats.

This study aimed to investigate the effects of metformin and forskolin independently and in combinations on the sperm quality parameters and sexual hormones of diabetic male rats. Fifty adult male rats were divided randomly into five identical groups, and diabetes mellitus was induced to the rats, except for the rats in the control group, using a high-fat diet and injection of Streptozotocin. Daily administration of metformin and forskolin independently and in combinations were performed for 8 weeks in different groups. Sperm quality parameters (including sperm count, morphology, sperm motility and Johnson score), testosterone, blood sugar level, Bax to Bcl-2 ratio mRNA expression level and oxidative stress levels were measured and compared between the investigated groups. Treating diabetic rats with metformin and forskolin resulted in significant improvement in sperm quality parameters, increased testosterone levels, reduced oxidative stress in blood and testicular tissue, and decreased blood sugar, and Bax to Bcl-2 ratio level. Although the combination of metformin with forskolin had a higher effect in some parameters such as testosterone levels compared to treatment with metformin or forskolin alone, this combination had not shown a synergistic effect in all the sperm quality parameters. Metformin and forskolin are effective anti-diabetic agents, which significantly improve the sperm quality and sexual hormone levels in diabetic rats. Combining metformin and gorskolin resulted in significantly better testosterone level and antioxidant activity in blood serum without significant effect on sperm quality of diabetic rats.

Inulin-Coated Iron Oxide Nanoparticles: A Theranostic Platform for Contrast-Enhanced MR Imaging of Acute Hepatic Failure.

The present study introduces a superparamagnetic nanocomposite, Fe-Si-In, as a T2 magnetic resonance imaging (MRI) contrast agent with a core of iron oxide nanoparticles and a nonporous silica inner shell/carboxymethyl inulin outer shell. Due to its core/shell properties, the structure characterization, biocompatibility, and performance in MRI, as well as its potential as a drug delivery system, were thoroughly evaluated. The results have shown that the synthesized nanocomposite possesses excellent biocompatibility and acceptable magnetization (Ms = 20 emu g-1). It also has the potential to be a nanocarrier for drug delivery purposes, as evidenced by the results of curcumin administration studies. The developed nanocomposite has shown excellent performance in MRI, while the in vitro relaxivity measurements reveal a stronger T2 relaxivity (r2 = 223.2 ms) compared to the commercial samples available in the market. Furthermore, the in vivo MRI studies demonstrate an excellent contrast between injured livers and normal ones in rats which again upholds the high performance of Fe-Si-In in MRI diagnostics.

Evaluation of bone regeneration in mandible large defect using undifferentiated adipose stem cells loaded on gelatin carrier: An animal model case study.

Background. Large mandibular defects are considered difficult reconstructive challenges for oral and maxillofacial surgeons. Cell therapy, as an alternative technique, might increase the speed of bone regeneration. This study aimed to investigate bone regeneration in large defects of dog mandibles using allogenic adipose-derived stem cells on gelatin foam as a cell carrier. Methods. The tissue engineering phase consisted of the sampling of adult dogs' adipose tissue that can easily be isolated from adipose stem cells (ASCs) of the dogs, ASCs were cultured in Dulbecco's Modified Eagle's Medium (DMEM, Gibco, USA) with low glucose, containing 10% fetal bovine serum (FBS) (Sigma, USA) and 1% penicillin-streptomycin (Gibco, USA), with the characterization of dog ASCs and gelatin-transplanted ASCs. Six dogs were included in this experimental study in the next step and randomly assigned to the treatment and control groups. The samples in both groups underwent surgery under general anesthesia to create uniform 3-cm bony defects. The samples in both groups were reconstructed with titanium reconstruction plates and screws. A large bone gap filled with ASCs (5×106 ) was seeded on gelatin (ASCs) in the treatment group. In the control group, bony defects were filled with a cell delivery carrier without ASCs. Six months after transplantation, the animals' mandibles were evaluated by CT scan imaging, and the results were quantified through the Hounsfield unit (HU). The data were analyzed with t-test. Results. Before transplantation, the nature of the stem cells was confirmed by the expression of CD44 and CD105 cell markers at 71.9% and 89.3%, respectively, and a lack of the CD45 cell marker expression at 2.2%. Evaluation of CT scan images showed significantly higher bone repair in the ASCs group (920.25±572.92 HU) than in the control group (-94.746± 08.42). Conclusion. The bone regeneration of the ASCs group was significantly higher than that in the control group.

PVP Surface-protected silica coated iron oxide nanoparticles for MR imaging application.

This paper proposed an engineered mesoporous silica-coated Fe3O4 nanoparticle, PVPMSFe, prepared by a sol-gel/surface-protected etching mechanism as an MRI T2 contrast agent. To this end, the structural characterization of the nanocomposite was performed by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) method, VSM, thermogravimetric analysis (TGA), TEM, FESEM, and energy-dispersive X-ray scanning electron microscopy (EDS). The findings show that the synthesized nanocomposite has a mesoporous structure with an average particle size of 11.8 nm and excellent magnetization properties. The biocompatibility of PVPMSFe was investigated by MTT assay and hemolysis assay of red blood cells and the results indicate that PVPMSFe has favorable biocompatibility. Besides, the effect of PVPMSFe was assessed with MRI relaxivity measurement (T2 signal). Regarding the in vitro MRI relaxivity measurements outputs (r2=144.4), PVPMSFe can attenuate the T2 signal of MRI, perfectly which makes it an efficient T2 contrast agent.

Corrigendum to "One-pot hydrothermal synthesis of a magnetic hydroxyapatite nanocomposite for MR imaging and pH-Sensitive drug delivery applications" [Heliyon 6 (9) (September 2020) e04928].

[This corrects the article DOI: 10.1016/j.heliyon.2020.e04928.].

One-pot hydrothermal synthesis of a magnetic hydroxyapatite nanocomposite for MR imaging and pH-Sensitive drug delivery applications.

Synthetic hydroxyapatite (HA) due to its high biocompatibility, anti-inflammatory properties, high stability, and a flexible structure in combination with magnetic nanoparticles has the strong potential to be used in modern medicine including tissue engineering, imaging, and drug delivery. Herein, a hydrothermal process was used to prepare magnetite nanoparticles dispersed on the hydroxyapatite nanorods with cetyltrimethylammonium bromide (CTAB) as a surfactant. Characterization study of the synthesized iron oxide-hydroxyapatite (IO-HA) nanocomposite was performed by FT-IR spectroscopy, X-ray powder diffraction, energy dispersive X-Ray analysis (EDX) for elemental mapping, transmission electron microscopy, and vibrating sample magnetometer. Then, the biocompatibility of the synthesized nanocomposite studied by 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay and hemocompatibility assay. Focus on this point, curcumin loaded IO-HA (Cur@IO-HA) was developed for exploring the pH-sensitivity of the drug carrier and then evaluating its cellular uptake. The in vitro efficacy of the synthesized nanocomposites as a magnetic resonance imaging (MRI) contrast agent was also investigated. Our results showed that IO-HA nanocomposite is non-cytotoxic and hemocompatible as well as a good pH-sensitive drug carrier and a favorable MRI T2 contrast agent. Comparing to the free curcumin, Cur@IO-HA displayed a good cellular uptake. Taking into account the above issues, IO-HA nanocomposite has the most potential for application as a theranostic MRI contrast agent.

Synthesis and characterisation of iron oxide nanoparticles conjugated with epidermal growth factor receptor (EGFR) monoclonal antibody as MRI contrast agent for cancer detection.

The aim of this study is to synthesise superparamagnetic iron oxide nanoparticles conjugated with anti-epidermal growth factor receptor monoclonal antibody (ANTI-EGFR-SPION) and investigate its physicochemical characterisation and biocompatibility as a targeted magnetic resonance imaging (MRI) contrast agent for the EGFR-specific detection in EGFR expressing tumour cells. These particles employed biocompatible polymers, poly(D,L-lactide-co-glycolide) (PLGA) and polyethylene glycol aldehyde (PEG-aldehyde), to increase the half-life of particles in circulation and reduce their side effects. The Fe3O4-loaded PLGA-PEG-aldehyde nanoparticles were prepared by a modified water-in-oil-in-water double emulsion method. The EGFR antibody was conjugated to the surface of SPIONs using the aldehyde-amine reaction. Synthesised conjugates (nanoprobes) were characterised using Fourier transform infrared spectrophotometry, dynamic light scattering, transmission electron microscopy images, and vibrating-sample magnetometery, and the results showed that the conjugation was successful. The mean diameter of nanoprobes was about 25 nm. These nanoprobes exhibited excellent water-solubility, stability, and biocompatibility. Meanwhile, MR susceptibility test proved that synthesised nanoprobes can be managed for negative contrast enhancement. The results of this study suggested the potential use of these nanoprobes for non-invasive molecular MRI in EGFR detection in the future.

Vertebrobasilar Artery Stroke as the Heralding Sign of Systemic Lupus Erythematosus.

Cerebral ischemia because of vertebrobasilar insufficiency (VBI) rarely presents as an initial sign within the systemic lupus erythematosus (SLE) population, and there are very few case reports supporting this manifestation. This report details 3 different patients with SLE who experienced VBI as an initial manifestation. Patient 1 was a 24-year-old female who developed a bilateral pontine lesion as a consequence of basilar artery stenosis. Patient 2 was a 34-year-old male with an acute ischemic lesion on the right side of his cerebellum and pons because of significant stenosis in the distal segment of the right vertebral artery. Patient 3 was a 37-year-old female, previously diagnosed with multiple sclerosis, with multiple lesions in her cerebellum and pons bilaterally. Further investigations within this case revealed severe stenosis of the left vertebral artery. The diagnosis of SLE was based on clinical presentations such as myalgia, skin rashes, ulcers, and fatigue along with relevant laboratory findings including positive anti ds-DNA antibody and depressed levels of complement C3 and C4 proteins. In young patients with multifocal ischemic lesions or infarcts in the posterior cerebral circulation system, physicians should investigate for less common etiologies such as SLE.

Synthesis and characterization of dextran coated magnetite nanoparticles for diagnostics and therapy.

INTRODUCTION: Expansion of efficacious theranostic systems is of pivotal significance for medicine and human healthcare. Magnetic nanoparticles (MNPs) are known as drug delivery system and magnetic resonance imaging (MRI) contrast agent. MNPs as drug carriers have attracted significant attention because of the delivery of drugs loaded onto MNPs to solid tumors, maintaining them in the target site by an external electromagnetic field, and subsequently releasing drugs in a controlled manner. On the other hand, it is believed that MNPs possess high potential as MRI contrast agents. The aim of this work was to payload curcumin into dextran coated MNPs and investigate their potential as theranostic systems for controlled drug delivery and MRI imaging. METHODS: MNPs were synthesized as a core and coated with dextran as polymeric shell to provide steric stabilization. Curcumin as anticancer drug was selected to be loaded into NPs. To characterize the synthesized NPs, various techniques (e.g., DLS, FESEM, FT-IR, XRD, and VSM) were utilized. In vitro drug release of curcumin was evaluated at 37˚C at the pH value of 5.4 and 7.4.The feasibility of employment of dextran coated MNPs as MRI contrast agents were also studied. RESULTS: Formulations prepared from dextran coated MNPs showed high loading (13%) and encapsulation efficiency (95%). In vitro release study performed in the phosphate-buffered saline (PBS, pH= 7.4, 5.4) revealed that the dextran coated MNPs possess sustained release behavior at least for 4 days with the high extent of drug release in acidic media. Vibrating sample magnetometer (VSM) analysis proved the superparamagnetic properties of the dextran coated MNPs with relatively high-magnetization value indicating that they were sufficiently sensitive to external magnetic fields as magnetic drug carriers. Furthermore, dextran coated MNPs exhibited high potential as T2 contrast agents for MRI. CONCLUSION: Based on our findings, we propose the dextran coated MNPs as promising nanosystem for the delivery of various drugs such as curcumin and MRI contrast agent.

The impact of polymer coatings on magnetite nanoparticles performance as MRI contrast agents: a comparative study.

BACKGROUND: Superparamagnetic iron oxide nanoparticles (SPIONs) are the most commonly used negative MRI contrast agent which affect the transverse (T2) relaxation time. The aim of the present study was to investigate the impact of various polymeric coatings on the performance of magnetite nanoparticles as MRI contrast agents. METHODS: Ferrofluids based on magnetite (Fe3O4) nanoparticles (SPIONs) were synthesized via chemical co-precipitation method and coated with different biocompatible polymer coatings including mPEG-PCL, chitosan and dextran. RESULTS: The bonding status of different polymers on the surface of the magnetite nanoparticles was confirmed by the Fourier transform infrared spectroscopy (FT-IR) and thermogravimetric analysis (TGA). The vibrating sample magnetometer (VSM) analysis confirmed the superparamagnetic behavior of all synthesized nanoparticles. The field-emission scanning electron microscopy (FE-SEM) indicated the formation of quasi-spherical nanostructures with the final average particle size of 12-55 nm depending on the type of polymer coating, and X-ray diffraction (XRD) determined inverse spinel structure of magnetite nanoparticles. The ferrofluids demonstrated sufficient colloidal stability in deionized water with the zeta potentials of -24.2, -16.9, +31.6 and -21 mV for the naked SPIONs, and for dextran, chitosan and mPEG-PCL coated SPIONs, respectively. Finally, the magnetic relaxivities of water based ferrofluids were measured on a 1.5 T clinical MRI instrument. The r2/r1 value was calculated to be 17.21, 19.42 and 20.71 for the dextran, chitosan and mPEG-PCL coated SPIONs, respectively. CONCLUSIONS: The findings demonstrated that the value of r2/r1 ratio of mPEG-PCL modified SPIONs is higher than that of some commercial contrast agents. Therefore, it can be considered as a promising candidate for T2 MRI contrast agent.