Effects of solid lipid nanocarrier containing methyl urolithin A by coating folate-bound chitosan and evaluation of its anti-cancer activity.

BACKGROUND: Nanotechnology-based drug delivery systems have received much attention over the past decade. In the present study, we synthesized Methyl Urolithin A-loaded solid lipid nanoparticles decorated with the folic acid-linked chitosan layer called MuSCF-NPs and investigated their effects on cancer cells. METHODS: MuSCF-NPs were prepared using a high-pressure homogenization method and characterized using FTIR, FESEM, DLS, and zeta potential methods. Drug encapsulation was assessed by spectrophotometry and its cytotoxic effect on various cancer cells (MDA-MB231, MCF-7, PANC, AGS, and HepG2) by the MTT method. Antioxidant activity was assessed by the ABTS and DPPH methods, followed by expression of genes involved in oxidative stress and apoptosis by qPCR and flow cytometry. RESULTS: The results showed the formation of monodisperse and stable round nanoparticles with a size of 84.8 nm. The drug loading efficiency in MuSCF-NPs was reported to be 88.6%. MuSCF-NPs exhibited selective cytotoxicity against MDA-MB231 cells (IC50 = 40 µg/mL). Molecular analysis showed a significant increase in the expression of Caspases 3, 8, and 9, indicating that apoptosis was occurring in the treated cells. Moreover, flow cytometry results showed that the treated cells were arrested in his SubG1 phase, confirming the pro-apoptotic effect of the nanoparticles. The results indicate a high antioxidant effect of the nanoparticles with IC50 values ​​of 45 µg/mL and 1500 µg/mL against ABTS and DPPH, respectively. The reduction of catalase gene expression confirmed the pro-oxidant effect of nanoparticles in cancer cells treated at concentrations of 20 and 40 µg/mL. CONCLUSIONS: Therefore, our findings suggest that the MuSCF-NPs are suitable candidates, especially for breast cancer preclinical studies.

Anti-Cancer and Anti-Oxidant Effects of Fenoferin-loaded Human Serum Albumin Nanoparticles Coated with Folic Acid-bound Chitosan.

BACKGROUND: Several diseases, including cancer, can be effectively treated by altering the nanocarrier surfaces so that they are more likely to be targeted. OBJECTIVE: This study aimed to prepare human albumin (HSA) nanoparticles containing Fenoferin (FN) modified with folic acid (FA) attached to Chitosan (CS) to improve its anti-cancer properties. METHODS: Nanoparticles were first synthesized and surface modified. Their physicochemical properties were assessed by different methods, such as FESEM, FTIR, and DLS. In addition, the percentage of drug encapsulated was measured by indirect method. Besides evaluating the cytotoxic effects of nanoparticles using the MTT assay, the antioxidant capacity of FN-HSA-CS-FA was assessed using the ABTS and DPPH methods. Nanoparticles were also investigated for their anti-cancer effects by evaluating the expression of apoptosis and metastasis genes. RESULTS: Based on this study, FN-HSA-CS-FA was 165.46 nm in size, and a uniform dispersion distribution was identified. Particles were reported to have a zeta potential of +29 mV, which is within the range of stable nanoparticles. Approximately 75% of FN is encapsulated in nanoparticles. Cytotoxic assay determined that liver cancer cells were most sensitive to treatment with an IC50 of 144 µg/ml. Inhibition of free radicals by nanoparticles is estimated to have an IC50 value of 195.23 and 964 µg/ml, for ABTS and DPPH, respectively. In the treatment with nanoparticles, flow cytometry results of arresting the cells in the SubG1 phase and real-time qPCR results indicated increased expression of caspases-3, caspase-8, and caspase-9 genes. CONCLUSION: According to this study, synthesized nanoparticles inhibited free radicals and activated apoptosis in liver cancer cells, and the capability of these nanoparticles to inhibit cancer cells was also confirmed. This formulation can, therefore, be used in preclinical studies to test the efficacy of the drug.

Study the Anticancer Properties of Thymol-Loaded PEGylated Bovine Serum Albumin Nanoparticles Conjugated with Folic Acid.

Phenolic compounds such as Thymol have an effective role in suppressing cancer, however, their low solubility in aqueous solution has limited their use. This study aimed to prepare Thymol (TY)-loaded bovine serum albumin (BSA) nanoparticles surface-modified with polyethylene glycol (PEG) conjugated with folic acid (FA) and evaluate their inhibitory activity on cancer cells. The TY-BSA-PEG-FA was characterized using DLS, FESEM, and FTIR. The encapsulation efficiency (EE) was evaluated indirectly by using UV absorption. The antioxidant property of nanoparticles was evaluated by 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing ability of plasm (FRAP) methods. The effects of nanoparticles against cancer cells were investigated by MTT, AO/PI, flow cytometry, and real-time qPCR methods. The results showed the spherical morphology of TY-BSA-PEG-FA with an average size of 70.0 nm, a PDI of 0.32, a zeta potential of -11.3 mV, and an EE of 89.0±2.3 %. The cytotoxicity effects of nanoparticles against all cell lines were in a concentration-dependent manner. AGS gastric cancer cells were reported to be the most vulnerable to treatment, while pancreatic cancer cells (PANC-1) and normal skin cells (HFF) would be the most resistant. The SubG1 phase arrest of about 66 % occurred at 85 µg/mL. An increase in apoptotic cells in fluorescent staining, along with decreased expression of Bcl-2 and increased expression of the BAX gene demonstrated the induction of apoptosis in treated cells. The powerful inhibitory effect of nanoparticles in inhibiting ABTS free radicals (IC50 =82 µg/mL) and DPPH free radicals (IC50 =844 µg/mL) and the ability to reduce iron ions indicated the antioxidant effects of TY-BSA-PEG-FA. Based on these results, the synthesized nanoparticles may be suitable for further investigation in the treatment of cancer, notably gastric cancer.

Synthesis and evaluation of biological effects of modified graphene oxide nanoparticles containing Lawson (Henna extract) on gastric cancer cells.

PURPOSE: Targeted Graphene Oxide (GO) nanoparticles can play an important role in the treatment of cancer by increasing cancer cell targeting. This study was conducted to synthesize GO nanoparticles functionalized with chitosan-folate (CS-FA) to deliver a natural product Lawsone (LA) for cancer treatment. METHODS: After characterization of the LA-GO-CS-FA, antioxidant activities of the nanoparticles were investigated by ABTS, DPPH, and FRAP tests. CAM assay was used to study the effect of nanoparticles on angiogenesis. The expression level of inflammatory and angiogenic genes in cells treated with nanoparticles was evaluated by real-time PCR. RESULTS: The findings demonstrated the formation of nanoparticles with a size of 113.3 nm, a PDI of 0.31, and a surface charge of + 11.07 mV. The percentages of encapsulation efficiency were reported at 93%. Gastric cancer cells were reported as the most sensitive to treatment compared to the control, and the gastric cancer cells were used to study gene expression changes. The anti-angiogenic effects of nanoparticles were confirmed by reducing the average number and length of blood vessels and reducing the height and weight of embryos in the CAM assay. The reducing the expression of genes involved in angiogenesis in real-time PCR was demonstrated. Nanoparticles displayed high antioxidant properties by inhibiting DPPH and ABTS radicals and reducing iron ions in the FRAP method. The reduction of pro-inflammatory genes in AGS cells which were treated with nanoparticles indicates the anti-inflammatory properties of nanoparticles. CONCLUSION: This study showed the efficacy of nanoparticles in inhibiting gastric cancer cells by relying on inhibiting angiogenesis.

Synthesis Folate-linked Chitosan-coated Quetiapine/BSA Nano-Carriers as the Efficient Targeted Anti-Cancer Drug Delivery System.

Quetiapine (QTP) has been known to suppress cancer progression in patients suffering from mental disorders. This study aimed to produce the folate-linked chitosan-coated quetiapine/BSA nano-carriers (FCQB-NCs) and evaluate their antioxidant, apoptotic, and anti-metastatic potentials on prostate, pancreas, colon, and breast cancer cell lines. The FCQB-NCs were designed, produced, and characterized using DLS, FESEM, FTIR, and Zeta potential techniques. The nano-carriers antioxidant activity was studied by applying ABTS, DPPH, and FRAP assays. The FCQB-NCs' cytotoxicity and apoptotic/metastatic properties were evaluated utilizing MTT assay and qPCR-based analysis for measuring the apoptotic (Nf-KB)/metastatic (MMP2, MMP9, and MMP13) gene expression, respectively. The AO/PI fluorescent cell staining, DAPI staining, and scratch assay methods were conducted to verify the apoptotic and anti-metastatic activities of FCQB-NCs. The 51-nm FCQB-NCs (PDI = 0.26) exhibited antioxidant activity and selectively decreased the MDA-MB-231 cancer cells' viability by inducing Nf-KB overexpression, which caused the apoptosis pathway activation. Moreover, the FCQB-NCs suppressed the MDA-MB-231 cells' metastatic activity by down-regulating the MMP2, MMP9, and MMP13 gene expression, verified by detecting the decreased migration rate. The FCQB-NCs selectively induced apoptosis and suppressed metastasis in the human breast cancer cell line, which can be attributed to the stepwise release of QTP in two primary (extra-cellular release) and secondary (intra-cellular release) phases. The efficient selective cytotoxic impact of FCQB-NCs can be due to the novel stepwise release mechanism of the FCQB-NCs based on the two-phase entrapment of QTP by BSA and chitosan molecules. Therefore, FCQB-NCs have the potential to be used as an efficient selective anti-breast cancer.

Evaluation of Serum Selenium and Copper Levels with Inflammatory Cytokines and Indices of Oxidative Stress in Type 2 Diabetes.

Type 2 diabetes mellitus (T2DM) is a metabolic and multifactorial disease in which inflammatory markers, oxidative stress, and certain trace elements seem to have an essential role. This study investigated the relationship between serum selenium and copper level with inflammatory cytokines and oxidative stress in T2DM.In this case-control study, 30 patients with T2DM and 30 healthy individuals were selected. Serum levels of copper and selenium were measured by atomic absorption spectrometry, and TNF-α and IL-6 and oxidative stress markers were measured by ELISA. The SPSS v.22 was used for data analysis and the significance level is less than 5%.The mean age of patients was 52.9 ± 10.4 years, and the control group was 48.5 ± 10.4 years. In this study, 53.3% were female, and 46.7% were male. The levels of BMI (p = 0.002), systolic pressure (p = 0.034), insulin, selenium, malondialdehyde, and glutathione peroxidase (p = 0.0001; each), insulin resistance, copper, and superoxide dismutase, IL6, and TNF-α (p = 0.001; each) in T2DM were significantly higher than the control group. While levels of lipid profile, uric acid, creatinine, and diastolic pressure were not significantly different between the two groups. Selenium and copper are related to insulin resistance, and their increasing levels are associated with increased levels of markers of oxidative stress and inflammatory cytokines (p < 0.05).Increased levels of copper and selenium are associated with T2DM and this increase is also associated with increased levels of TNF-α, IL-6, and oxidative stress in T2DM. Therefore, controlling these markers can lead us to control this disease better.

Assessment of the Relationship Between CD34 Antigen and Vitamin D Level and Insulin Resistance in Patients With Type 2 Diabetes.

Diabetes causes disorders in the performance of endothelial progenitor cells, and obesity and vitamin D deficiency are associated with endothelial dysfunction and cardiovascular disease. This case-control study investigated the relationship between serum CD34 antigen and vitamin D levels and insulin resistance in type 2 diabetes. The results showed that CD34 has a significant inverse relationship with BMI, A1C, fasting blood glucose, insulin resistance, and insulin levels and has a significant direct relationship with vitamin D levels. Both CD34 and vitamin D were found to be significantly associated with type 2 diabetes. The association between reduced CD34 and vitamin D levels with type 2 diabetes and increased insulin resistance suggests that these parameters may be helpful in assessing diabetes and predicting its complications.

Investigation of serum level relationship anti-glutamic acid decarboxylase antibody and inflammatory cytokines (IL1-ß, IL-6) with vitamins D in type 2 diabetes.

Background: Various factors are involved in the development and progression of diabetes mellitus, from cytokines and autoimmune destruction of pancreatic beta cells to vitamin D.This study aimed to investigate the association of serum levels of anti-glutamic acid decarboxylase (anti-GAD) and inflammatory cytokines with vitamin D in type 2 diabetes (T2D). Methods: This case-control study was performed on 30 patients with T2D and 30 healthy individuals in Mashhad hospitals in 2020. Lipid profile, creatinine, uric acid, FBS, HbA1c, and blood pressure were recorded. All study variables were measured, particularly serum vitamin D, anti-GAD, and inflammatory cytokine levels in diabetic patients, and the data were compared to those from healthy subjects by performing an appropriate statistical analysis. Results: Diabetic patients with a mean age of 52.9 ± 10.4 years, including 16 women and healthy individuals with a mean age of 48.5 ± 10.4 years, including 16 women, were studied. BMI level (P = 0.002), systolic blood pressure (P = 0.034), HbA1c, insulin, IL-6, IL1-ß, anti-GAD levels, and insulin resistance in diabetic patients were significantly higher than the control group (P = 0.001). The vitamin D level in the control group was significantly higher than in the case group (P = 0.0001). The results showed a significant direct relationship between IL-6, IL-1ß, and anti-GAD with HbA1c, FBS, insulin, and insulin resistance. However, there was a significant inverse relationship between IL-6, IL-1ß, and anti-GAD with vitamin D. Conclusions: Inflammatory cytokines and anti-GAD and vitamin D are associated with diabetes, and thus controlling these factors can help improve T2D.

Clinicopathological utility of miR-203a-3p in diagnosing colorectal cancer.

BACKGROUND: Biomarkers, such as microRNAs, are helpful in diagnosing colorectal cancer, regulating disease progression, predicting disease recurrence, and determining therapy success. This research aimed to look at the clinicopathological characteristics of serum miRNA-203a-3p expression in colorectal cancer patients. METHODS AND RESULTS: This case-control study was conducted on 43 patients with colorectal cancer and 43 healthy individuals. After RNA extraction, cDNA was synthesized. The expression of miR-203a-3p was measured using RT-qPCR. Demographic and histochemical data were extracted from patient documents. SPSS and GraphPad Prism software were used to analyze the data. The expression of miR-203a-3p in CRC patients was 2.39 times lower than in the control group (p < 0.0001). The miR-203a-3p expression was significantly lower in the CRC tumor stages, tumor grades, and lymph node metastasis compared to the control group (p < 0.0001 each). The ROC curves showed that the AUC was 0.73, and the best cut-point based on the Youden index was 0.3954, 0.7105, 0.5087, and 0.4868 for detecting colorectal cancer (p = 0.0002), tumor grade (p = 0.006), tumor stage (p = 0.001), and lymph node metastasis (p = 0.0011) compared to the control group, respectively. The binary logistic regression analysis was performed on the correlation between BMI, smoking, and cancer inheritance with miR-203a-3p in cancer and control groups. CONCLUSION: This study's findings revealed that serum miR-203a-3p is a fair non-invasive molecular biomarker for diagnosing and progressing tumor grade, tumor stage, and lymph node metastasis in colorectal cancer. However, further research with higher statistical numbers is needed to strengthen the correlation and be used for diagnostic applications.

The Anti-Breast Cancer Effects of Green-Synthesized Zinc Oxide Nanoparticles Using Carob Extracts.

BACKGROUND: The use of nanoparticles synthesized by the green method to treat cancer is fairly recent. The aim of this study was to evaluate cytotoxicity, apoptotic and anti-angiogenic effects and the expression of involved genes, of Zinc Oxide Nanoparticles (ZnO-NPs) synthesized with Carob extracts on different human breast cancer cell lines. METHODS: ZnO-NPs were synthesized using the extracts of Carob and characterized with various analytical techniques. The MCF-7 and MDA-MB231 cells were treated at different times and concentrations of ZnO-NPs. The cytotoxicity, apoptosis, and anti-angiogenic effects were examined using a series of cellular assays. Expression of apoptotic genes (Bax and Bcl2) and anti-angiogenic genes, Vascular Endothelial Growth Factor (VEGF) and its Receptor (VEGF-R) in cancer cells treated with ZnO-NPs were examined with Reverse Transcriptionquantitative Polymerase Chain Reaction (RT-qPCR). The anti-oxidant activities of ZnO-NPs were evaluated by ABTS and DPPH assay. RESULTS: Exposure of cells to ZnO-NPs resulted in a dose-dependent loss of cell viability. The IC50 values at 24, 48, and 72 hours were 125, 62.5, and 31.2µg/ml, respectively (p<0.001). ZnO-NPs treated cells showed, in fluorescent microscopy, that ZnO-NPs are able to upregulate apoptosis and RT-qPCR revealed the upregulation of Bax (p<0.001) and downregulation of Bcl-2 (p<0.05). ZnO-NPs increased VEGF gene expression while decreasing VEGF-R (p<0.001). The anti-oxidant effects of ZnO-NPs were higher than the control group and were dose-dependent (p<0.001). CONCLUSION: ZnO-NPs synthetized using Carob extract have the ability to eliminate breast cancer cells and inhibit angiogenesis, therefore, they could be used as an anticancer agent.

Clinical Evaluation of the Diagnostic Role of MicroRNA-155 in Breast Cancer.

AIM: Biochemical markers, including microRNAs (miRs), may facilitate the diagnosis and prognosis of breast cancer. This study was aimed at assessing serum miR-155 expression in patients with breast cancer and receptors. METHODS: This case-control study was conducted on 36 patients with breast cancer and 36 healthy individuals. After RNA extraction from the patient's serum, cDNA was synthesized. The expression of miR-155 was measured using RT-qPCR. Demographic and histochemical data were extracted from patient documents. Data were analyzed using the Statistical Package for the Social Sciences (SPSS) software. RESULTS: The mean age of subjects in breast cancer and control groups was 47.64 ± 8.19 and 47.36 ± 7.52 years, respectively. The serum miR-155 expression was higher in the cancer group (1.68 ± 0.66) compared to the control group (p < 0.0001). There was a significant relationship between serum miR-155 expression and the tumor grade (p < 0.001), tumor stage (p < 0.001), and tumor size (p < 0.001) of the patients. However, no relationship between miR-155 expression and the presence of lymph node involvement (p = 0.15), HER2 (p = 0.79), Ki-67 (p = 0.9), progesterone receptor (p = 0.54), and estrogen receptors (p = 0.84) was found. The ROC curve analysis showed that the AUC was 0.89 (77.78% sensitivity and 88.89% specificity), and the cutoff was 1.4 (Youden index: 0.6667) for detecting breast cancer. CONCLUSION: The findings of this study revealed that serum miR-155 may serve as a potential noninvasive molecular biomarker for breast cancer diagnosis and can help predict the grade of the disease.

Evaluation of the Diagnostic Properties of Serum hsa-miR-223-5p in the Detection of Gastric Cancer: A Case-Control Study.

BACKGROUND: MicroRNAs (miRs) are a group of small non-coding and single-stranded RNAs of 18 to 25 nucleotides. The study of microRNAs is one of the new ways to detect cancer. In this study, the serum expression of miR-223 in patients with GC was measured and compared with the control group. METHODS: This case-control study was conducted on 39 patients with GC and 39 control subjects who visited the Reza Radiotherapy and Oncology Center, Mashhad, Iran, due to gastrointestinal complaints. The demographic information was collected, and the serum levels of miR-223 were measured using the real-time PCR technique in all study subjects. The association between the GC of miR-223 and tumor staging and cancer progression was assessed. RESULTS: The miR-223 expression in GC patients was 3.10-fold higher than that of the control group (p<0.0001). The miR-223 expression was significantly higher in the GC stages and grades compared to the control group (p<0.0001 each). However, there was no significant effect for age, smoking, and gender on miR- 223 expression in GC and controls. At the optimal cutoff value of 0.7436, the maximal sensitivity of 89.74% and specificity of 84.62% were achieved for miR-223 (p<0.001). The sensitivity and specificity for miR-223 for differentiating low grades from high grade were 92.31% and 73.08% (p=0.0003), and for differentiating low stages from the high stage was 81.82% and 39.29% respectively (p=0.696). CONCLUSION: This study revealed that miR-223 could be considered as a non-invasive diagnostic marker in the early diagnosis of GC.

Putative mechanism for anticancer properties of Ag-PP (NPs) extract.

One of the most important challenges in treating cancer is the invasion and the angiogenesis of cancer cells. The synthesis of green nanoparticles (NPs) and their use in therapeutic fields is one of the most effective methods with minimal side effects in cancer treatment. In this study, cytotoxic and anti-angiogenic effects of silver NPs (AgNPs) coated with palm pollen extract [Ag-PP(NPs)] were evaluated. For this purpose, the cells were treated with NPs and then were subjected to trypan blue testing (48 h). Then, the cancer invasion was evaluated by the scratch procedure and the expressions of the vascular endothelial growth factor (VEGF) and its receptor (VEGF-R) genes were estimated using real-time PCR assay. Also, the angiogenesis effect of the NPs was investigated with chick chorioallantoic membrane (CAM) assay. The Ag-PP(NPs) induced cytotoxicity on MCF7 cells. The findings also showed that Ag-PP(NPs) inhibit invasive cancer cells and reduce the expression of VEGF and VEGF-R and significantly reduced the number and vessels lengths and the lengths and weights of the embryos in CAM assay. Ag-PP(NPs) with the induction of cytotoxic effects, metastatic inhibition and anti-angiogenesis properties should be considered as an appropriate option for treatment of cancer.