Increased radiosensitivity of melanoma cells through cold plasma pretreatment mediated by ICG.

Radiation therapy (RT) is the primary treatment for many cancers, but its effectiveness is reduced due to radioresistance and side effects. The study aims to investigate an emerging treatment for cancer, cold atmospheric plasma (CAP), as a selectable treatment between cancerous and healthy cells and its role in the occurrence of photodynamic therapy (PDT) utilizing indocyanine green (ICG) as a photosensitizer. We examined whether the efficiency of radiotherapy could be improved by combining CAP with ICG. The PDT effect induced by cold plasma irradiation and the radiosensitivity of ICG were investigated on DFW and HFF cell lines. Then, for combined treatment, ICG was introduced to the cells and treated with radiotherapy, followed by cold plasma treatment simultaneously and 24-h intervals. MTT and colony assays were used to determine the survival of treated cells, and flow cytometry was used to identify apoptotic cells. Despite a decrease in the survival of melanoma cells in CAP, ICG did not affect RT. Comparing the ICG + CAP group with CAP, a significant reduction in cell survival was observed, confirming the photodynamic properties of plasma utilizing ICG. The treatment outcome depends on the duration of CAP. The results for healthy and cancer cells also confirmed the selectivity of plasma function. Moreover, cold plasma sensitized melanoma cells to radiotherapy, increasing treatment efficiency. Treatment of CAP with RT can be effective in treating melanoma. The inclusion of ICG results in plasma treatment enhancement. These findings help to select an optimal strategy for a combination of plasma and radiotherapy.

Molecular MR Imaging of Prostate Cancer by Specified Iron Oxide Nanoparticles With PSMA-11 Peptides: A Preclinical Study.

BACKGROUND: Prostate-specific membrane antigen (PSMA) can provide a prostate cancer (PCa) detection approach in positron emission tomography (PET) using Food and Drug Administration (FDA)-approved PSMA-11 peptide. There are some studies evaluated magnetic-nanoprobes for PSMA detection by MRI, using non-FDA-approved ligands including antibodies or peptides, which are not as specific as PSMA-11. PURPOSE: To assess targeted iron oxide nanoparticles (IONPs) by PSMA-11 peptides as a potential specific nano-molecular probes to investigate a PSMA+ PCa-xenograft model by MRI. STUDY TYPE: Prospective. ANIMAL MODEL: Twenty male C57BL6 nude mice induced subcutaneously PSMA+ LNCaP cell line tumor. FIELD STRENGTH/SEQUENCE: 1.5 T, T2 -W Fast Spin echo and T2 *-W Gradient echo. ASSESSMENT: Coated IONPs with Carboxymethylated-dextran (DNPs) and with bovine serum albumin (BNPs), as well as, targeted DNPs with PSMA-11-HYNIC peptide (TDNPs) and targeted BNPs with PSMA-11-HBED peptide (TBNPs) were injected intravenously with dose 2.8 mg Fe/kg. Coronal T2 -W and the T2 *-W images were obtained before and 4 hours and 6 hours post-injection. Signal intensity (SI) and relative signal enhancement (RSE) were computed in two- and three-dimensional analyses. Histological analysis of tumors was evaluated, and the Fe distribution within the body based on atomic absorption spectroscopy was calculated. STATISTICAL TESTS: One-way ANOVA followed by Tukey's multiple comparison test, Paired-samples T-test, P < 0.05 was considered significant. RESULTS: A reduction in T2 -W SI was achieved as 22 ± 7%, 59 ± 3%, 65 ± 5%, and 78 ± 3% respectively for BNPs, TBNPs, DNPs, and TDNPs 6 hours post-injection. The most difference between targeted and non-targeted groups was observed at 6 hours for PSMA-11-HBED, and at 4 hours for PSMA-11-HYNIC. RSE indicated 88.6 ± 3.1% and 80.7 ± 3.2% enhanced contrast between tumor and muscle region for TBNPs and TDNPs on T2 *-W images. CONCLUSIONS: Both TBNPs and TDNPs are promising novel nano-molecular probes for PSMA+ PCa tumor detection. The injection dose of non-targeted IONPs can be reduced by using targeted nanoprobes three times for BNPs and two times for DNPs. EVIDENCE LEVEL: 1 TECHNICAL EFFICACY: Stage 1.

Histopathological graded liver lesions: what role does the IVIM analysis method have?

PURPOSE: This study aims to investigate three different image processing methods on quantitative parameters of IVIM sequence, as well as apparent diffusion coefficients and simple perfusion fractions, for benign and malignant liver tumors. MATERIALS AND METHODS: IVIM images with 8 b-values (0-1000 s/mm2) and 1.5 T MRI scanner in 16 patients and 3 healthy people were obtained. Next, the regions of interest were selected for malignant, benign, and healthy liver regions (50, 56, and 12, respectively). Then, the bi-exponential equation of the IVIM technique was fitted with two segmented fitting methods as well as one full fitting method (three methods in total). Using the segmented fitting method, diffusion coefficient (D) is fixed with a mono-exponential equation with b-values that are greater than 200 s/mm2. The perfusion fraction (f) can then be calculated by extrapolating, as the first method, or fitting simultaneously with the pseudo-diffusion coefficient (D*) as the second method. In the full fitting method, as the third method, all IVIM parameters were obtained simultaneously. The mean values of parameters from different methods were compared in different grades of lesions. RESULTS: Our results indicate that the image processing method can change statistical comparisons between different groups for each parameter. The D value is the only quantity in this technique that does not depend on the fitting process and can be used as an indicator of comparison between studies (P < 0.05). The most effective method to distinguish liver lesions is the extrapolated f method (first method). This method created a significant difference (P < 0.05) between the perfusion parameters between benign and malignant lesions. CONCLUSION: Using extrapolated f is the most effective method of distinguishing liver lesions using IVIM parameters. The comparison between groups does not depend on the fitting method only for parameter D.

Chronic obstructive pulmonary disease and asthma: mesenchymal stem cells and their extracellular vesicles as potential therapeutic tools.

Chronic lung diseases, such as chronic obstructive pulmonary disease (COPD) and asthma, are one of the most frequent causes of morbidity and mortality in the global. COPD is characterized by progressive loss of lung function through inflammation, apoptosis, and oxidative stress caused by chronic exposure to harmful environmental pollutants. Airway inflammation and epithelial remodeling are also two main characteristics of asthma. In spite of extensive efforts from researchers, there is still a great need for novel therapeutic approaches for treatment of these conditions. Accumulating evidence suggests the potential role of mesenchymal stem cells (MSCs) in treatment of many lung injuries due to their beneficial features including immunomodulation and tissue regeneration. Besides, the therapeutic advantages of MSCs are chiefly related to their paracrine functions such as releasing extracellular vesicles (EVs). EVs comprising exosomes and microvesicles are heterogeneous bilayer membrane structures loaded with various lipids, nucleic acids and proteins. Due to their lower immunogenicity, tumorigenicity, and easier management, EVs have appeared as favorable alternatives to stem cell therapies. Therefore, in this review, we provided an overview on the current understanding of the importance of MSCs and MSC-derived EVs from different sources reported in preclinical and clinical COPD and asthmatic models.

Ultrasensitive and label free electrochemical immunosensor for detection of ROR1 as an oncofetal biomarker using gold nanoparticles assisted LDH/rGO nanocomposite.

In the present article, we developed a highly sensitive label-free electrochemical immunosensor based on NiFe-layered double hydroxides (LDH)/reduced graphene oxide (rGO)/gold nanoparticles modified glassy carbon electrode for the determination of receptor tyrosine kinase-like orphan receptor (ROR)-1. In this electrochemical immunoassay platform, NiFe-LDH/rGO was used due to great electron mobility, high specific surface area and flexible structures, while Au nanoparticles were prepared and coated on the modified electrodes to improve the detection sensitivity and ROR1 antibody immobilizing (ROR1Ab). The modification procedure was approved by using cyclic voltammetry and differential pulse voltammetry based on the response of peak current to the step by step modifications. Under optimum conditions, the experimental results showed that the immunosensor revealed a sensitive response to ROR1 in the range of 0.01-1 pg mL-1, and with a lower limit of quantification of 10 attogram/mL (10 ag mL-1). Furthermore, the designed immunosensor was applied for the analysis of ROR1 in several serum samples of chronic lymphocytic leukemia suffering patients with acceptable results, and it also exhibited good selectivity, reproducibility and stability.

Investigation of Specific Targeting of Triptorelin-Conjugated Dextran-Coated Magnetite Nanoparticles as a Targeted Probe in GnRH+ Cancer Cells in MRI.

In recent years, the conjugation of superparamagnetic iron oxide nanoparticles (SPIONs), as tumor-imaging probes for magnetic resonance imaging (MRI), with tumor targeting peptides possesses promising advantages for specific delivery of MRI agents. The objective of the current study was to design a targeted contrast agent for MRI based on Fe3O4 nanoparticles conjugated triptorelin (SPION@triptorelin), which has a great affinity to the GnRH receptors. The SPIONs-coated carboxymethyl dextran (SPION@CMD) conjugated triptorelin (SPION@CMD@triptorelin) were synthesized using coprecipitation method and characterized by DLS, TEM, XRD, FTIR, Zeta, and VSM techniques. The relaxivities of synthetized formulations were then calculated using a 1.5 Tesla clinical magnetic field. MRI, quantitative cellular uptake, and cytotoxicity level of them were estimated. The characterization results confirmed that the formation of SPION@CMD@triptorelin has been conjugated with a suitable size. Our results demonstrated the lack of cellular cytotoxicity of SPION@CMD@triptorelin, and it could increase the cellular uptake of SPIONs to MDA-MB-231 cancer cells 6.50-fold greater than to SPION@CMD at the concentration of 75 µM. The relaxivity calculations for SPION@CMD@triptorelin showed a suitable r 2 and r 2/r 1 with values of 31.75 mM-1·s-1 and 10.26, respectively. Our findings confirm that triptorelin-targeted SPIONs could provide a T 2-weighted probe contrast agent that has the great potential for the diagnosis of GnRH-positive cancer in MRI.

IL-10-producing B cells play important role in the pathogenesis of recurrent pregnancy loss.

Interleukin (IL)-10-producing B cells are recently known for their regulatory function in several disorders. However, the possible role of these cells remains unclear in recurrent pregnancy loss (RPL) pathogenesis. Total B cells from 24 RPL patients with cellular immune abnormalities, as well as that of 25 normal pregnant women were cultured and stimulated by Toll Like Receptor (TLR) agonists (CpG oligodeoxynucleotides (ODN) and imiquimod). Then, the frequency of IL-10+ CD19+ B cells was found out using flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to assess the levels of IL-10 in supernatant medium and serum of stimulated B cells, as well as those of several serum autoantibodies. Real-Time PCR method was carried out for determining the IL-10 expression level and specific genes transcripts. RPL patients indicated a lower proportion of IL-10+ CD19+ B cells, and reduced levels of IL-10 in both serum and supernatant of the culture medium of the stimulated B cells. According to the results, total IgG levels was greater in serum of RPL patients in comparison with healthy pregnant women. Similarly, the percentage of these cells was negatively correlated with serum total IgG levels and the number of miscarriages. The expression levels of the mRNA of programmed death-ligand 1 (PD-L1) and IL-10 were lower in RPL patients, while those of x binding protein 1 (XBP-1), interferon regulatory factor 4 (IRF4), and B lymphocyte-induced maturation protein 1 (BLIMP1) were significantly increased. These observations indicated that the reduction in the population of peripheral blood IL-10-synthesizing B cells may prompt RPL pathogenesis, suggesting suppressive effects of these cells on autoantibody production and successful pregnancy outcomes.

Regenerative potential of Wharton's jelly-derived mesenchymal stem cells: A new horizon of stem cell therapy.

Umbilical cord Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) have recently gained considerable attention in the field of regenerative medicine. Their high proliferation rate, differentiation ability into various cell lineages, easy collection procedure, immuno-privileged status, nontumorigenic properties along with minor ethical issues make them an ideal approach for tissue repair. Besides, the number of WJ-MSCs in the umbilical cord samples is high as compared to other sources. Because of these properties, WJ-MSCs have rapidly advanced into clinical trials for the treatment of a wide range of disorders. Therefore, this paper summarized the current preclinical and clinical studies performed to investigate the regenerative potential of WJ-MSCs in neural, myocardial, skin, liver, kidney, cartilage, bone, muscle, and other tissue injuries.

Kinetic and thermodynamic study of c-Met interaction with single chain fragment variable (scFv) antibodies using phage based surface plasmon resonance.

Mesenchymal epithelial transition factor (c-Met) has been recently regarded as an attractive target for the treatment of cancer. Our previous study showed that c-Met-specific single chain fragment variables (scFvs) can be considered as a promising therapy for cancer, however, their molecular interaction with c-Met protein have not been assessed. Accordingly, in the current study we aim to evaluate the kinetic and thermodynamic properties of c-Met interaction with these scFvs as anticancer agents by means of surface plasmon resonance (SPR) technique. Phage-scFvs were immobilized on the 11-mercaptoundecanoic acid gold chips after carboxylic groups activation by N-ethyl-N-(3-diethylaminopropyl) carbodiimide/N-hydroxysuccinimide and, then the c-Met binding to each scFvs (ES1, ES2, and ES3) at different concentrations (ranging from 20 to 665 µM) was explored. Kinetic studies revealed that ES1 has the highest affinity (KD = 3.36 × 10-8) toward its target at 25°C. Calculation of thermodynamic parameters also showed positive values for enthalpy and entropy changes, which was representative of hydrophobic forces between c-Met and ES1. Furthermore, the positive value of Gibbs free energy indicated that c-Met binding to ES1 was enthalpy-driven. Taken together, we concluded that produced ES1 can be applied as promising scFv-based therapy for diagnosis or targeting of c-Met in various cancers.

Immunological and oxidative stress biomarkers in Ankylosing Spondylitis patients with or without metabolic syndrome.

Ankylosing Spondylitis (AS) is a chronic inflammatory disorder of the spine and sacroiliac joints with unidentified etiology closely associated with metabolic syndrome (MetS). Recent studies have reported that immunological and oxidative stress factors are implicated in AS pathogenesis. The aim of this study was to investigate the oxidative and immunological factors in AS patients with or without MetS compare to control group. Real-Time PCR measured expression level of cytokines, transcription factors and related miRNAs. In addition, Th17 and Treg frequencies and cytokines secretion were evaluated by flowcytometry and ELISA methods, respectively. The oxidative stress biomarkers were also assessed with biochemical methods. In AS patients with MetS, higher Th17 and lower Treg frequency were observed. Increased levels of NF-kB and AP-1 mRNA expression were seen in AS patients with MetS (p = 0.0263 and p = 0.0104, respectively). MiR-146a and miR-223 were significantly decreased (p = 0.0005, p = 0.0161, respectively) and increase in miR-21 (p = 0.0002) was observed in AS patients with MetS compared to AS patients without MetS. Additionally, the secretion of TNF-α (p = 0.0167), IL-1ß (p = 0.303), CCL2 (p = 0.0254), CCL3 (p = 0.0119), CXCL8 (p = 0.0364), adiponectin (p = 0.0183) and the levels of SOD (p = 0.0421), NO (p = 0.0451) and CAT (p = 0.0128) were increased in AS patients with MetS. We were not observed significant differences in TOS and GPX levels between studied groups. The higher levels of oxidative stress and immunological inflammatory markers in AS patients with MetS provide further evidences on the oxidative stress and immunological relationship in these patients.

Human umbilical cord mesenchymal stem cell-derived extracellular vesicles: A novel therapeutic paradigm.

Mesenchymal stem cells (MSCs) have been revealed to hold great potential for the development of new treatment approaches for various diseases. However, the clinical use of these cells is limited due to their tumorigenic effects. The therapeutic benefits of MSCs are largely dependent on paracrine factors including extracellular vesicles (EVs). EVs are nano-sized bilayer membrane structures containing lipids, microRNAs and proteins which play key roles in cell-to-cell communications. Because of their lower immunogenicity, tumorigenicity, and easier management, EVs have emerged as a new promising alternative to whole-cell therapy. Therefore, this paper reviews current preclinical studies on the use of EVs derived from human umbilical cord MSCs (hucMSCs) as a therapeutic approach in treatment of several diseases including neurological, cardiovascular, liver, kidney, and bone diseases as well as the cutaneous wound, inflammatory bowel disease, cancers, infertility, and other disorders.

Biosensors and nanobiosensors for rapid detection of autoimmune diseases: a review.

This review (with 77 refs.) describes the progress that has been made in biosensors for the detection of autoimmune diseases, mainly via detection of autoantibodies. In addition, specific proteins, cytokines and ions have also been introduced as promising diagnostic biomarkers. Following an introduction into the various kinds of autoimmune diseases, we first discuss the state of the art in respective electrochemical biosensors and nanobiosensors (with subsections on amperometric, impedimetric, voltammetric and photoelectrochemical methods). The next large chapter covers optical methods (with subsections on electrochemiluminescence, fluorescence and surface plasmon resonance). We then make a critical comparison between commercially available kits used for detection of autoimmune diseases with the established biosensors. Several Tables are also presented that give an overview on the wealth of methods and nanomaterials. Finally, in the conclusion part, we summarize the current status, addresse present issues, and give an outlook on potential future opportunities. Graphical abstractSchematic representation of various developed optical and electrochemical biosensors and nanobiosensors for rapid detection of autoimmune diseases nanobiosensors for rapid detection of autoimmune diseases which could significantly prevent irreversable tissue damages and increse the quality of life in these patients.

The role of IL-10-producing B cells in repeated implantation failure patients with cellular immune abnormalities.

There are a few data of the role of B cells in RIF pathogenesis. Accordingly, the objective of the current study was to determine the role of IL-10-producing B cells in RIF. Twenty-three RIF women with cellular immune abnormalities and 25 normal controls were enrolled in this experiment. Isolated naïve B cells from peripheral blood of the subjects were cultured in vitro, divided into two parts and activated by CpG ODN and imiquimod as TLR agonists. Afterwards, the number of CD19+ IL-10+ B cells was evaluated by flow cytometry and their related IL-10 cytokine level was assessed by ELISA. The mRNA expression levels of related genes in just CPG stimulated B cell population were also analyzed using real-time PCR. RIF patients exhibited a decreased level of the cells (P = 0.014, P = 0.023, respectively) and IL-10 cytokine (P = 0.009, P = 0.045, respectively) in both CPG and imiquimod stimulated B cell groups. IL-10 serum level was also lower in these patients (P = 0.0014). Additionally, we found a negative relationship between the frequency of these cells with the number of failed ET and total IgG titers in RIF patients. The mRNA levels of IL-10-producing B cells related genes (IL-10 and PD-L1) was also significantly lower in RIF women, whereas the expression of plasma cells-associated transcriptional factors (BLIMP1, IRF4, and XBP1) was higher. Summing up the obtained results, we concluded that peripheral blood IL-10-producing B cells down-regulation might result in RIF pathogenesis. It is further suggested that these cells can suppress autoantibody generation and contribute to a successful implantation.

Application of various optical and electrochemical aptasensors for detection of human prostate specific antigen: A review.

Early stage detection of prostate cancer, one of the main causes of mortality among men, is of great importance for better treatment of the patients. Prostate specific antigen (PSA) is a glycoprotein which has been considered as the most potential serological biomarker for the detection of prostate cancer. Among the various techniques employed for PSA detection, aptamer-based biosensors (aptasensors) have achieved notable attention because of their unique features and great potentials as diagnostic tools. A variety of strategies such as integration of nanomaterials (NMs) into the structure of aptasensors have also been applied for enhancing the sensitivity of PSA detection. This article reviews recent advances in various optical and electrochemical aptasensors used for PSA detection.

A novel nano-superparamagnetic agent for photodynamic and photothermal therapies: An in-vitro study.

BACKGROUND: In this study, iron oxide nanoparticles (SPIONs) were synthesized and coated by GA (SG) and then SG was encapsulated by ICG (SGI). After identifying specifications and cytotoxicity of the agents, the potential of SGI for photodynamic therapy (PDT) and photothermal therapy (PTT) was studied. METHODS: An SGI size of 12-13 nm was determined by TEM images and its zeta potential was measured at -23.8 ± 5.8 mV. MCF-7 and HT-29 cells were exposed to a non-coherent light source at a wavelength of 730 nm and a range of 3.9-124.8 J/cm2 under two different concentrations of agents. The viability of treated cells was determined via MTT assay. To analyze the effects of different irradiation conditions, some indices such as Coefficient of Light Effect, Synergism Index, Addition Ratio, Treatment Efficacy and ED50 were defined. RESULTS: Cell survival at the highest power of irradiation in the absence of any agent was decreased to 93% and 73% for HT-29 and MCF-7, respectively. In both cell lines, the cellular survival dropped by increasing the light source intensity. The maximum cell death recorded for SG, ICG and SGI was 63 ± 2%, 63 ± 2% and 21 ± 2% for MCF-7 cells and 67 ± 2%, 78 ± 1% and 53 ± 1% for HT-29 cells, respectively. CONCLUSION: SGI had a significant photodynamic and photothermal effect on cells. This is a promising outcome, which can help enhance the effectiveness of a minimally invasive treatment. Moreover, SPIONs can be used to apply magnetic hyperthermia or act as a contrast agent in MRI images.

The synergistic effect of photodynamic therapy and photothermal therapy in the presence of gold-gold sulfide nanoshells conjugated Indocyanine green on HeLa cells.

BACKGROUND: Photodynamic therapy (PDT) and photothermal therapy (PTT) are two known optical remedies of cancer. PTT can be combined with other therapies. One of the limitations of optical therapies is the penetration of light into biological tissues, which reduces its effectiveness due to usage of photosensitizers and PTT agents, which are absorbed in the NIR region that provides the maximum penetration. For instance, Indocyanine green (ICG) serves as a photosensitizer and Gold nanostructures as agents for PTT. GGS is a gold nanoshell with two absorption peaks in the NIR and visible regions. The aim of this study is to evaluate the synergistic effect of PDT and PTT in the presence of GGS conjugated with ICG. METHODS: After synthesizing GGS, ICG was conjugated with GGS. The specifications and cytotoxicity of agents were identified. Cells were irradiated by an 808nm laser with or without the agents and three laser outputs were achieved, with each having four different exposure times. The viability of treated cells was determined via MTT assay. RESULT: The irradiation of the laser did not produce any significant effect by itself or in the presence of GGS. The maximum cell death recorded for GGS, ICG and GGS-ICG were 15±7%, 50±3% and 31±3% respectively. ICG and GGS-ICG differs significantly for exposures higher than 2250J/cm2. CONCLUSION: The conjugate was provided through a simple process and a greater chemical stability compared to GGS was achieved. Moreover, it induced a stronger photodynamic and photothermal effect on the cells. This is a promising result which can help enhance the effectiveness of a minimally invasive treatment.