Advanced Pharmaceutical Bulletin
Year:2022 | Volume:12 | Issue:3|Papers Count:22

Cytokines, as protein biomarkers, have essential functions in the diagnosis, identification, and healing of a broad range of syndromes. For the specific and accurate monitoring of immune conditions, which change rapidly throughout the duration of disease, sophisticated sensors for detecting cytokines are essential and will assist in clinical testing and studies of various diseases. The present manuscript briefly discusses fundamental principles applied to the development of tools for cytokine detection and new biomarker development. The latest developments in the technologies for highly sensitive and multiplexed cytokine quantification, with current detection capabilities across a broad, vibrant array, are also discussed. Finally, nanomaterial-based cytokine sensors, currently considered new approaches, are presented from the perspective of optimizing the sensitivity and multiplexity of cytokine detection.

The novel coronavirus (COVID-19) that first appeared in December 2019, subsequently named severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) is rapidly spreading as a global pandemic. Following infection by SARS-CoV-2, systemic inflammatory response mediated by the release of large amounts of mediators including IL-6, IL-1b, TNFα,and IL-2R in severe infected patients. . . .

Chronic wounds have made a challenge in medical healthcare due to their biofilm infections, which reduce the penetrance of the antibacterial agents in the injury site. In infected wounds, the most common bacterial strains are Staphylococcus aureus and Pseudomonas aeruginosa. Biofilm disruption in chronic wounds is crucial in wound healing. Due to their broad-spectrum antibacterial properties and fewer side effects, anti-biofilm peptides, especially bacteriocins, are promising in the healing of chronic wounds by biofilm destruction. This study reviews the effects of antimicrobial and anti-biofilm agents, including bacteriocins and protease enzymes as a novel approach, on wound healing, along with analyzing the molecular docking between a bacterial protease and biofilm components. Among a large number of anti-biofilm bacteriocins identified up to now, seven types have been registered in the antimicrobial peptides (AMPs) database. Although it is believed that bacterial proteases are harmful in wound healing, it has recently been demonstrated that these proteases like the human serine protease, in combination with AMPs, can improve wound healing by biofilm destruction. In this work, docking results between metalloprotease from Paenibacillus polymyxa and proteins of S. aureus and P. aeruginosa involved in biofilm production, showed that this bacterial protease could efficiently interact with biofilm components. Infected wound healing is an important challenge in clinical trials due to biofilm production by bacterial pathogens. Therefore, simultaneous use of proteases or anti-biofilm peptides with antimicrobial agents could be a promising method for chronic wound healing.

Tissue engineering is a novel regenerative approach in the medicinal field that promises the regeneration of damaged tissues. Moreover, tissue engineering involves synthetic and natural biomaterials that facilitate tissue or organ growth outside the body. Not surprisingly, the demand for polymer-based therapeutical approaches in skin tissue defects has increased at an effective rate, despite the pressing clinical need. Among the 3D scaffolds for tissue engineering and regeneration approaches, hydrogel scaffolds have shown significant importance for their use as 3D cross-linked scaffolds in skin tissue regeneration due to their ideal moisture retention property and porosity biocompatibility, biodegradable, and biomimetic characteristics. In this review, we demonstrated the choice of ideal biomaterials to fabricate the novel hydrogel scaffolds for skin tissue engineering. After a short introduction to the bioactive and drug-loaded polymeric hydrogels, the discussion turns to fabrication and characterisation techniques of the polymeric hydrogel scaffolds. In conclusion, we discuss the excellent wound healing potential of stem cell-loaded hydrogels and Nano-based approaches to designing hydrogel scaffolds for skin tissue engineering.

The development of carbon-based nanomaterials has extensively facilitated new discoveries in various fields. Carbon nanotube-based nanocomposites (CNT-based nanocomposites) have lately recognized as promising biomaterials for a wide range of biomedical applications due to their unique electronic, mechanical, and biological properties. Nanocomposite materials such as silver nanoparticles (AgNPs), polymers, biomolecules, enzymes, and peptides have been reported in many studies, possess a broad range of antibacterial activity when incorporated with carbon nanotubes (CNTs). It is crucial to understand the mechanism which governs the antimicrobial activity of these CNT-based nanocomposite materials, including the decoupling individual and synergistic effects on the cells. In this review, the interaction behavior between microorganisms and different types of CNT-based nanocomposites is summarized to understand the respective antimicrobial performance in different conditions. Besides, the current development stage of CNT-based nanocomposite materials, the technical challenges faced, and the exceptional prospect of implementing potential antimicrobial CNT-based nanocomposite materials are also discussed.

Purpose: blood-brain barrier (BBB) is made of specialized cells that are responsible for the selective passage of substances directed to the brain. The integrated BBB is essential for precise controlling of the different substances passage as well as protecting the brain from various damages. In this article, we attempted to explain the role of liver X receptor (LXR) in maintaining BBB integrity as a possible drug target. Methods: In this study, various databases, including PubMed, Google Scholar, and Scopus were searched using the following keywords: blood-brain barrier, BBB, liver X receptor, and LXR until July, 2020. Additionally, contents close to the subject of our study were surveyed. Results: LXR is a receptor the roles of which in various diseases have been investigated. LXR can affect maintaining BBB by affecting various ways such as ATP-binding cassette transporter A1 (ABCA1), matrix metalloproteinase-9 (MMP9), insulin-like growth factor 1 (IGF1), nuclear factor-kappa B (NF-κ, B) signaling, mitogen-activated protein kinase (MAPK), tight junction molecules, both signal transducer and activator of transcription 1 (STAT1), Wnt/β,-catenin Signaling, transforming growth factor beta (TGF-β, ) signaling, and expressions of Smad 2/3 and Snail. Conclusion: LXR could possibly be used either as a target for drug delivery to brain tissue or as a target for maintaining the BBB integrity in different diseases,thereby the drug will be conducted to tissues, other than the brain. If it is verified that only LXRα,is necessary for protecting BBB, some specific LXRα,ligands must be found and then used in medication.

Immunotherapy has become a prominent strategy for the treatment of cancer. A method that improves the immune system’, s ability to attack a tumor (Enhances antigen binding). Targeted killing of malignant cells by adoptive transfer of chimeric antigen receptor (CAR) T cells is a promising immunotherapy technique in the treatment of cancers. For this purpose, the patient’, s immune cells, with genetic engineering aid, are loaded with chimeric receptors that have particular antigen binding and activate cytotoxic T lymphocytes. That increases the effectiveness of immune cells and destroying cancer cells. This review discusses the basic structure and function of CAR-T cells and how antigenic targets are identified to treat different cancers and address the disadvantages of this treatment for cancer.

Schizophrenia is a neuropsychiatric disorder mainly affecting the central nervous system (CNS), presented with auditory and visual hallucinations, delusion and withdrawal from society. Abnormal dopamine levels mainly characterise the disease,various theories of neurotransmitters explain the pathophysiology of the disease. The current therapeutic approach deals with the systemic administration of drugs other than the enteral route, altering the neurotransmitter levels within the brain and providing symptomatic relief. Fluid biomarkers help in the early detection of the disease, which would improve the therapeutic efficacy. However, the major challenge faced in CNS drug delivery is the blood-brain barrier (BBB). Nanotherapeutic approaches may overcome these limitations, which will improve safety, efficacy, and targeted drug delivery. This review article addresses the main challenges faced in CNS drug delivery and the significance of current therapeutic strategies and nanotherapeutic approaches for a better understanding and enhanced drug delivery to the brain, which improve the quality of life of schizophrenia patients.

Prostate cancer (PCa) is one of the leading diseases in men all over the world caused due to over-expression of prostate-specific membrane antigen (PSMA). Currently, the detection and targeting of PCa is one of the major challenges in the prostate gland. Therefore, Bruton tyrosine kinase inhibitor molecules like ibrutinib (Ibr) loaded with nanomaterials like multiwalled carbon nanotubes (MWCNTs), which has good physico-chemical properties may be the best regimen to treat PCa. In this strategy, the chemically modified MWCNTs have excellent ‘, Biosensing’,properties makes it easy for detecting PCa without fluorescent agent and thus targets particular site of PCa. In the present study, Ibr/MWCNTs conjugated with T30 oligonucleotide may selectively target and inhibit PSMA thereby reduce the over-expression in PCa. Hence, the proposed formulation design can extensively reduce the dosage regimen without any toxic effect. Additionally, the present hypothesis also revealed the binding mode of Ibr in the catalytic pocket of PSMA by in silico method. Therefore, we presume that if this hypothesis proves correct, it becomes an additional novel tool and one of the conceivable therapeutic options in treating PCa.

Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system (CNS). It is an auto-immune disorder. Its usual symptoms are unique to each person. In MS lesions vast fractions of pyruvate molecules are instantly transformed into lactate. This reprogramming mechanism of glycolysis is known as the Warburg effect. MS has no efficient treatment yet. Hence, there is a requirement for profitable immunomodulatory agents in MS. Probiotics perform as an immunomodulator because they regulate the host’, s immune responses. Its efficacy gets enhanced for an extended period when it combines with prebiotics. In this review, we focus on the metabolic alterations behind the MS lesions via the Warburg effect, and also suggesting, the combined efficacy of prebiotics and probiotics for the effective treatment of MS without side effects. The Warburg effect mechanism intensifies the infiltration of activated T-cells and B-cells into the CNS. It provokes the inflammation process on the myelin sheath. The infiltration of immune cells can be inhibited by the combination therapy of probiotics and prebiotics. By this review, we can recommend that the idea of this combinational therapy can do miracles in the treatment of MS in the future.

Purpose: Photothermal therapy (PTT) is a procedure that converts laser beam energy to heat so can disturb tumor cells. Carbon nanotubes (CNTs) have unique properties in absorption optical energy and could change optical power into heat in PTT procedures. Additionally, titanium dioxide (TiO2) nanoparticles (NPs) have a unique feature in absorbing and scattering light. Therefore, these mentioned NPs could play a synergistic role in the PTT method. Methods: CNTs and TiO2 NPs were injected into the melanoma tumor sites of cancerous mice. Then sites were excited using the laser beam (λ,= 808 nm, P = 2 W, and I = 4 W/cm2). Injected NPs caused hyperthermia in solid tumors. Tumor size assay, statistical analysis, and histopathological study of the treated cases were performed to assess the role of mentioned NPs in PTT of murine melanoma cancer. Results: The results showed that CNTs performed better than TiO2 NPs in destroying murine melanoma cancer cells in animals. Conclusion: The present study compared the photothermal activity of excited CNTs and TiO2 NPs in cancer therapy at the near-infrared spectrum of light. Tumors were destroyed selectively because of their weakened heat resistance versus normal tissue. PTT of malignant melanoma through CNTs caused remarkable necrosis into the tumor tissues versus TiO2 NPs.

Purpose: To overcome the challenges caused by the use of conventional ophthalmic dosage forms such as the fast elimination of the drug from the surface of the eye, in this study, dual drug-loaded nanofibers were developed for sustained ophthalmic delivery of gentamicin (GNT) and methylprednisolone (MP). Moreover, the solvent effects, polymer mixtures, and method of preparation on the release profile of the prepared nanofibers, were evaluated. Methods: The nanofibers were prepared using polycaprolactone (PCL), poly (lactic-co-glycolic acid) (PLGA), and polyvinyl alcohol (PVA) using electrospinning technique. Thereafter, seven optimized formulations were developed with different solvent mixtures and polymer concentrations using various electrospinning methods. The physicochemical and mechanical properties of nanofibers were also evaluated, and the morphology of formulations was observed. The antibacterial efficacy was investigated and the in vitro release amounts of GNT and MP from nanofibers were estimated using the bioassay and ultraviolet-visible (UV-Vis) spectroscopy. Results: The developed G1, G4, G5, G6, and G7 had suitable mechanical properties and morphologies with diameter ranging between 70-350 nm. The 1: 1 v/v ratio of DMF/DCM in the solvent mixture and using core-shell technique for the preparation, formed nanofibers with more favorable release profiles. The optimized formulations indicated sustained-release manner for both drugs during 3-9 days and the antibacterial efficacy against Staphylococcus aureus. Conclusion: Among all the prepared formulations, the nanofiber with core-shell structure possessed the best sustained-release profiles of GNT and MP. The obtained results suggest that these nanofibers have a potential to be used as an insert in the eye for long-term release of the drug.

Purpose: Free radicals such as hydroxyl and peroxide are contributing factors to neuronal destruction in cerebral ischemia. Alpha-lipoic acid (ALA) is one of the potent known antioxidants. Preparation of ALA niosomes allows IV injection and can increase bioavailability and penetration into the central nervous system (CNS). Methods: Film hydration method was used to prepare different niosomes composed of Span®, , Tween®, , and cholesterol at different molar ratio. ALA and niosome-forming compounds were dissolved in chloroform, before removing the organic solvent by rotary evaporator. Animals were randomly divided into four groups: Sham, control group, intravenous (IV) injection of empty niosomes plus intraperitoneal (IP) injection of ALA solution, and finally, IV injection of ALA niosomes. Rats were subjected to deep anesthesia before inducing cerebral ischemia, then, their internal common carotid arteries were clamped for 15 min and reperfusion was done for 30 min. Niosomal ALA was injected intravenously just before declamping. Results: Mean volume diameter of the prepared niosomes was between 4. 36 ±,0. 82 and 19. 95 ±,1. 21 μ, m in different formulations. Encapsulation efficiency percent (EE%) of ALA in the selected formulation, Span60/Tween60/cholesterol (35: 35: 30 molar ratio), was 94. 5 ±,0. 2, and 59. 27 ±,5. 61% of ALA was released after 4h. In the niosomal group, the rate of reduction in complications of cerebral ischemia such as histopathologic changes and acute damage (from score 3 to 1) in CNS was higher than other groups. Conclusion: The obtained results show that niosomes can be used as effective drug delivery systems for ALA in cerebral ischemia.

Purpose: Currently, several disorders including burns, trauma, excisional and diabetic wounds, and bedsores threaten the human health. Application of mesenchymal stem cells (MSCs) is recommended for treatment of skin disorders. However, because of oxidative stress and inflammation after skin injury, survival of transplanted MSCs is low which in turn negatively affects the efficiency of the MSCs-based therapy. In an attempt to address the aforementioned challenge and introducing a novel potential therapeutic strategy, we employed combination therapy by lipocalin 2 (Lcn2)-engineered MSCs and a Metadichol (an inverse agonist of vitamin D receptor (VDR)) nanogel in a rat model of excisional wound. Methods: First, human umbilical cord MSCs (hUC-MSCs) was transfected by a recombinant plasmid encoding Lcn2 gene. Next, a combination of Metadichol nanogel and the engineered MSCs was co-applied on wound in rat model of excision injury. Finally the improvement of wound healing in experimental groups was evaluated by photography and histological assessments (hematoxylin and eosin staining). Results: Our findings revealed that the repair rate was higher in the group received combination therapy comparing to control groups. Notably, Metadichol+Lcn2-MSCs showed significantly higher wound contraction rate compared to control group at all time points (P value < 0. 001). Furthermore, wound repair rate was 95% 14 days after surgery, and 100% after 21 days in the treatment groups. Our results also revealed that the combination therapy improved and accelerated the wound healing process. Conclusion: Our findings suggest a novel potential therapeutic strategy i. e. Lcn2-engineered MSCs and Metadichol for wound healing. However, further preclinical and clinical studies are required.

Purpose: This research introduces a polymeric nanosphere as a new dispersive solid phase extraction (DSPE) adsorbent for the extraction of methylphenidate (MPH) from urine and its high performance liquid chromatography (HPLC) analysis. Methods: Polymeric nanosphere is a kind of copolymeric network obtained by copolymerization of an ionic liquid monomer and styrene in the presence of vinyltriethoxysilane and 2-hydroxyethylmethacrylate. HPLC coupled with ultra violet detector was applied for the determination and quantification of MPH. Dominant parameters in extraction were modified by the one-parameter-at-a-time method. The results are as follow: 10 mg of polymeric nanospheres (PNS), 400 μ, L of acetonitrile (ACT), 5 mL of urine with the pH value of 9, and the extraction and desorption times of 2 and 5 minutes, respectively, which can be selected as the optimum extraction conditions. Results: Calibration curve was plotted through optimized conditions, and the proposed method was validated. The results demonstrated that the method presented linearity in the concentration range of 30-1200 ng/mL. Selectivity, matrix effect and metabolites interference effect were investigated and the method presented no obvious interference effect during the analysis run time. Repeatability, limit of detection (LOD) and limit of quantification (LOQ) values of the method can be reported in this section as well. The method showed satisfactory results with 98. 8% relative recovery in the analysis of positive urine samples. Conclusion: The findings convinced the applicability of the introduced method for DSPE and HPLC analysis of the positive urine samples in different laboratories.

Purpose: 5-Fluorouracil (5FU) and Fe3O4 nanoparticles were encapsulated in core-shell polycaprolactone (PCL)/chitosan (CS) nanofibers as a multi-mode anticancer system to study drug release sustainability. The structure of the core-shell drug delivery system was also optimized according to drug release behavior by artificial intelligence. Methods: The core-shell nanofibers were electrospun by a coaxial syringe. Artificial neural network (ANN) was used for function approximation to estimate release parameters. A genetic algorithm was then used for optimizing the structure. Chemical assay of the optimized sample was performed by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDX). vibration sample magnetometer (VSM) test was conducted to measure the real amount of loaded magnetic nanoparticles. HepG2 cell cytotoxicity was studied and the results for the optimized samples with and without Fe3O4 after 72 hours were reported. Results: Feeding ratio of sheath to core and the amount of CS, Fe3O4, and 5FU had a statistical effect on nanofibers diameters, which were 300-450 nm. The drug loading efficiency of these nanofibers was 65-86%. ANN estimated the release parameters with an error of 10%. The temperature increased about 5. 6°, C in the alternative magnetic field (AMF) of 216 kA. m-1~300 kHz and 4. 8°, C in the AMF of 154 kA. m-1~400 kHz after 20 minutes. HepG2 cell cytotoxicity for the optimized samples with and without Fe3O4 after 72 hours were 39. 7% and 38. 8%, respectively. Conclusion: Since this core-shell drug release system was more sustainable compared to the blend structure despite the low half-life of 5FU, it is suggested to utilize it as post-surgical implants for various cancer treatments such as liver or colorectal cancer in the future. This system is capable of providing chemotherapy and hyperthermia simultaneously.

Purpose: Production of functional recombinant antibody fragments in the periplasm of E. coli is a prerequisite step to achieve sufficient reagent for preclinical studies. Thus, the cost-effective and lab-scale production of antibody fragments demands the optimization of culture conditions. Methods: The culture conditions such as temperature, optical density (OD600) at induction, induction time, and IPTG concentration were investigated to optimize the functional expression of a phage-derived scFv molecule using a design of experiment (DoE). Additionally, the effects of different culture media and osmolyte supplements on the expression yield of scFv were examined. Results: The developed 2FI regression model indicated the significant linear effect of the incubation temperature, the induction time, and the induction OD600 on the expression yield of functional scFv. Besides, the statistical analysis indicated that two significant interactions of the temperature/induction time and the temperature/induction OD600 significantly interplay to increase the yield. Further optimization showed that the expression level of functional scFv was the most optimal when the cultivation was undertaken either in the TB medium or in the presence of media supplements of 0. 5 M sorbitol or 100 mM glycine betaine. Conclusion: In the present study, for the first time, we successfully implemented DoE to comprehensively optimize the culture conditions for the expression of scFv molecules in a phage antibody display setting, where scFv molecules can be isolated from a tailor-made phage antibody library known as “, Human Single Fold scFv Library I. ”,

Purpose: The hypoxia in solid tumors is associated with the resistance to chemo/radiotherapy. Hypoxia-inducible factor-1 (HIF-1) plays a key role in cell remodeling to hypoxia. Therefore, the inhibition of HIF-1 accumulation is considered a hopeful strategy for the treatment of cancer. Here, we aimed to evaluate the geno-and cytotoxicity properties of sclareol, a natural bicyclic diterpene alcohol, on A549 cells in CoCl2-induced hypoxia. Methods: The cytotoxicity and apoptosis-inducing properties of sclareol on the A549 cell were evaluated using MTT assay and Annexin V/PI staining, respectively in hypoxia. DAPI staining, DNA ladder, and comet assay were used to evaluate the genotoxicity. Further, the qPCR technique was employed to assess the expression of HIF-1α, , HIF-1β, , and downstream target genes (GluT1, and Eno1). Finally, the level of HIF-1α,protein was evaluated through Western blotting in sclareol-treated cells in hypoxia. Results: The inhibitory concentration (IC50) of sclareol against A549 cells was 8 μ, g/mL at 48 hours in hypoxia. The genotoxicity of sclareol was confirmed in the cells treated with sclareol in hypoxia. Sclareol induced ~46% apoptosis and also necrosis in the hypoxic condition. The qPCR analyses showed an enhanced suppression of HIF-1α, , HIF-1β, , GluT1, and Eno1 due to the sclareol treatment in the hypoxia. Moreover, protein quantification analysis showed dosedependently degradation of HIF-1α,in hypoxia upon treatment with sclareol. Conclusion: The results obtained here indicate that sclareol possesses dose-dependent cytotoxicity effects against A549 cells in hypoxia through inhibition of HIF-1α,protein accumulation, increasing cell sensitivity to intracellular oxygen levels, and disruption of cell adaptation to hypoxia.

Purpose: This study aimed to challenge the anticancer potency of pentagamavunone-1 (PGV-1) and obtain a new compound (Chemoprevention-Curcumin Analog 1. 1, CCA-1. 1) with improved chemical and pharmacological properties. Methods: CCA-1. 1 was prepared by changing the ketone group of PGV-1 into a hydroxyl group with NaBH4 as the reducing agent. The product was purified under preparative layer chromatography and confirmed with HPLC to show about 93% purity. It was tested for its solubility, stability, and cytotoxic activities on several cancer cells. The structure of the product was characterized using 1HNMR, 13C-NMR, FT-IR, and HR-mass spectroscopy. Results: Molecular docking analysis showed that CCA-1. 1 performed similar or better interaction to NF-κ, B pathway-related signaling proteins (HER2, EGFR, IKK, ER-alpha, and ER-beta) and reactive oxygen species (ROS) metabolic enzymes (NQO1, NQO2, GSTP1, AKC1R1, and GLO1) compared with PGV-1, indicating that CCA-1. 1 exhibits the same or better anticancer activity than PGV-1. CCA-1. 1 also showed better solubility and stability than PGV-1 in aqueous solution at pH 1. 0–, 7. 4 under light exposure at room temperature. The cytotoxic activities of CCA-1. 1 against several (10) cancer cell lines revealed the same or better potency than PGV-1. Conclusion: In conclusion, CCA-1. 1 performs better chemical and anticancer properties than PGV-1 and shows promise as an anticancer agent with high selectivity.

Purpose: Although the complex structure of acute lymphoblastic leukemia (ALL) and involvement of diverse pathways in its pathogenesis have put an obstacle in the way of efficient treatments, identification of strategies to manipulate the genome of neoplastic cells has made the treatment prospective more optimistic. Methods: To evaluate whether the transduction of apoptin __a gene encoding a protein that participates in the induction of apoptosis__ could reduce the survival of leukemic cells, we generated recombinant lentivirus expressing apoptin, and then, MTT assay, flow cytometric analysis of DNA content, western blotting, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were applied. Results: Transduction of apoptin into different leukemic cells was coupled with the reduction in the viability and proliferative capacity of the cells. Among all tested cell lines, Nalm-6 and C8166 were more sensitive to the anti-leukemic property of apoptin. Moreover, we found that the transduction of apoptin in the indicated cell lines not only induced G2/M cell cycle arrest but also induced apoptotic cell death by altering the balance between pro-and anti-apoptotic target genes. The efficacy of apoptin transduction was not limited to these findings, as we reported for the first time that the overexpression of this gene could potentiate the anti-leukemic property of pan PI3K inhibitor BKM120. Conclusion: The results of this study showed that the transduction of apoptin into lymphoblastic leukemia cell lines induced cytotoxic effects and enhanced therapeutic value of PI3K inhibition,however, further investigations are demanded to ascertain the safety and the efficacy of apoptin transduction in patients with ALL.

Purpose: Neuroinflammation was indicated in the pathophysiology of Alzheimer’, s disease (AD). Previous reports have also signified that spironolactone has anti-inflammatory effects. Therefore, the aim of this study was to assess the modulatory effects of spironolactone on neuroinflammation and memory loss in a rat model of AD. Methods: The β,-amyloid protein fragment 25-35 (Aβ, ) was injected in the dorsal hippocampus (5 μ, g/2. 5 μ, L each side) of male Sprague-Dawley rats for four consecutive days to induce memory impairment. Animals have intraperitoneally received spironolactone (10, 25, or 50 mg/kg, N = 6/ group) or vehicle for 14 days. The passive inhibitory avoidance and the novel recognition tests were used for memory evaluation. Neuroinflammation was assessed by measuring the level of Iba1 protein, a marker of microglial activation, using western immunoblotting. Results: Different doses of spironolactone showed no significant changes in latency times and discriminations ratios in passive inhibitory avoidance and novel recognition tests, respectively, as compared to vehicle. However, spironolactone-treated groups showed significantly lower Iba1 protein levels in comparison to the vehicle-treated group (P < 0. 01). Conclusion: Spironolactone had a modulatory effect on neuroinflammation through a repressive effect on microglial activation with no valuable effect on memory improvement in a rat model of AD. The findings of this study suggest that Aβ,-induced memory loss may not be directly linked to microglial activation. Spironolactone may be a potential candidate to be examined in other neuroinflammatory disorders.

Purpose: The aim of this study was to evaluate the protective effect of conditioned medium derived from human adipose mesenchymal stem cells (CM-hADSCs) on C28I2 chondrocytes against oxidative stress and mitochondrial apoptosis induced by high glucose (HG). Methods: C28I2 cells were pre-treated with CM-hADSCs for 24 hours followed by HG exposure (75 mM) for 48 hours. MTT assay was used to assess the cell viability. Reactive oxygen species (ROS) and lipid peroxidation were determined by 2, 7-dichlorofluorescein diacetate (DCFHDA) and thiobarbituric acid reactive substances (TBARS) assays, respectively. Expressions of glutathione peroxidase 3 (GPX 3), heme oxygenase-1 (HO-1), and NAD(P)H quinone dehydrogenase 1 (NQO1) were analyzed by RT-PCR. Finally, western blot analysis was used to measure Bax, Bcl-2, cleaved caspase-3, and Nrf-2 expression at protein levels. Results: CM-hADSCs pretreatment mitigated the cytotoxic effect of HG on C28I2 viability. Treatment also markedly reduced the levels of ROS, lipid peroxidation, and augmented the expression of HO-1, NQO1, and GPx3 genes in HG-exposed group. CM-ADSCs enhanced Nrf-2 protein expression and reduced mitochondrial apoptosis through reducing Bax/Bcl-2 ratio and Caspase-3 activation. Conclusion: MSCs, probably through its paracrine effects, declined the deleterious effect of HG on chondrocytes. Hence, therapies based on MSCs secretomes appear to be a promising therapeutic approaches to prevent joint complications in diabetic patients.