The Effect of a Casein and Gluten-Free Diet on the Epigenetic Characteristics of FoxP3 in Patients With Hashimoto's Thyroiditis.

Background Hashimoto's thyroiditis (HT) is an autoimmune thyroid disease characterized by inflammation and dysfunction of the thyroid gland, resulting in hypothyroidism, it results in impaired thyroid hormone generation and mimics hypothyroidism. The disease involves complex interactions among genetic, environmental, and epigenetic factors, particularly affecting the regulation of T regulatory (Treg) cells, including CD4 + foxp3 + T cells. Treg cells, defined as CD4 + T cells, rely on the expression of the foxp3 transcription factor, which is crucial for their development and differentiation. Disruptions in this regulation can lead to immune dysregulation and potential proinflammatory responses. The study focuses on investigating the impact of dietary patterns on the epigenetic changes in the foxp3 gene, a key player in the development of HT. The primary aim was to evaluate how eliminating gluten and casein proteins from dietary regimens may influence the methylation levels of the foxp3 gene, considering the potential link between these dietary components and the triggering of autoimmune diseases. Methods An epigenetic analysis of the foxp3 gene in HT patients who were strictly following a dietary plan compared with the control group. For the epigenetic study, a methylation analysis experiment was conducted.  Results Our findings revealed a notable reduction in foxp3 gene methylation levels among HT patients who adhered to a diet excluding casein and gluten. The control maintained normal dietary guidelines and showed no significant alterations in methylation levels. Discussion The laboratory values showed a decrease in methylation levels of the foxp3 gene, with statistical significance indicated as *p<0.005, **p<0.001, ***p<0.0001, suggesting a potential enhancement in its expression which could have profound implications for immune system regulation. Disruptions in the foxp3 pathway are crucial in the development of autoimmune disorders, where altered activity hinders the regulation of T cell (Treg) development, ultimately contributing to conditions like HT disease. These findings imply that nutritional interventions, especially for individuals with HT, could potentially be a strategy for mitigating autoimmunity through epigenetic mechanisms.

Electrospun nanofiber mats caged the mammalian macrophages on their surfaces and prevented their inflammatory responses independent of the fiber diameter.

Poly-ε-caprolactone (PCL) has been widely used as biocompatible materials in tissue engineering. They have been used in mammalian cell proliferation to polarization and differentiation. Their modified versions had regulatory activities on mammalian macrophages in vitro. There are also studies suggesting different nanofiber diameters might alter the biological activities of these materials. Based on these cues, we examined the inflammatory activities and adherence properties of mammalian macrophages on electrospun PCL nanofibrous scaffolds formed with PCL having different nanofiber diameters. Our results suggest that macrophages could easily attach and get dispersed on the scaffolds. Macrophages lost their inflammatory cytokine TNF and IL6 production capacity in the presence of LPS when they were incubated on nanofibers. These effects were independent of the mean fiber diameters. Overall, the scaffolds have potential to be used as biocompatible materials to suppress excessive inflammatory reactions during tissue and organ transplantation by caging and suppressing the inflammatory cells.

Anti-inflammatory activity of benidipine hydrochloride in LPS-activated mammalian macrophages.

Benidipine hydrochloride (BH), a medication frequently used by the hypertension patients, acts as a calcium channel blocker. However, its effects on the macrophages have not been investigated thus far. Our goal was investigating the effect of the benidipine hydrochloride to modulate the J774.2 murine macrophage cells inflammatory activity. Our results suggest that in the absence of a standard stimulating agent (LPS) BH did not stimulate the macrophages to produce pro-inflammatory IL-12p40, TNF-α, GM-CSF and IL-6 cytokines. However, when BH was administrated to the cells in the presence of LPS as stimulating agent, it reduced the production of these pro-inflammatory cytokines. Therefore, it had anti-inflammatory activity. At the clinical setting this study suggests that BH can be utilized as hypertension drug that can suppress the inflammation associated with it.

Photodynamic anti-inflammatory activity of meso­aryl substituted porphyrin derivative on mammalian macrophages.

BACKGROUND: Our group focused on a meso­aryl substituted porphyrin molecule for its photodynamic anti-inflammatory activities on the mammalian macrophages. MATERIALS AND METHODS: The porphyrine derivative previously synthesized in this study was synthesized and characterized by 1H NMR. We then examined their immunomodulatory activities based on the changes in the pro-inflammatory cytokine production levels after LPS stimulation in dark and light activated conditions. RESULTS: Our results suggest that porphyrin derivative had anti-inflammatory photodynamic activity in vitro at subtoxic concentrations. Our study aims to pave a way for anti-inflammatory photodynamic therapy application in the inflammatory and autoimmune disorders. Most of the studies either focus on photodynamic cytotoxicity of the porphyrin derivatives to suppress the inflammation or porphyrin derivatives' anti-inflammatory activity without the photodynamic activation. CONCLUSION: Our future studies will focus on the generation and in vitro as well as in vivo characterization of the porphyrin derivatives with anti-inflammatory photodynamic therapy applications. In this way, novel drug candidates that would have lower side effects can be generated for the patients.

Synthesis, anti-inflammatory activity, inverse molecular docking, and acid dissociation constants of new naphthoquinone-thiazole hybrids.

Chronic Inflammation is associated with various types of diseases that involves pro-inflammatory cytokines like IL-6 and TNF-α. High costs and serious side effects of available anti-inflammatory/immunomodulatory drugs led us to design new compounds with promising anti-inflammatory activities. Many drugs and biologically important compounds involve naphthoquinone and thiazole moieties in their core structures. Thereby, here we report the synthesis, characterization and anti-inflammatory activities of new naphthoquinone thiazole hybrids by reaction of naphthoquinone acyl thioureas with various α-bromoketone derivatives. The position of NO2 group in one of the phenyl rings of naphthoquinone thiazole hybrids was changed while different substituents were introduced at the para position of the second phenyl ring. All compounds were tested for potential immunomodulatory effect. No inflammatory cytokines were observed in the absence of LPS stimulant. On the other hand, they had promising anti-inflammatory immunomodulatory activities by being able to decrease the production of the pro-inflammatory cytokines (TNF-α and IL-6) in the LPS-stimulated cells. In an effort to find the possible mechanism of action, several enzymes involved in signalling pathways that play critical roles in inflammatory responses were screened in silico. Subsequent to inverse molecular docking approach, PI3K was predicted be the potential target. The docked complexes of the most potent compounds 5g and 5i were subjected to molecular dynamics simulation to assess the binding stability of the igands with the putative target. Acid dissociation constants (pKa) of the products were also determined potentiometrically.

Paroxetine's effect on the proinflammatory cytokine stimulation and intracellular signaling pathways in J774.2 cells.

Paroxetine is extensively utilized in the management of depressive and anxious conditions. Paroxetine works by increasing serotonin levels in nerve cells in the brain. However, limited information is available regarding the direct effects of paroxetine on macrophage cells. Macrophages are a type of leukocytes involved in the body's immune response, playing a crucial role in combating infections. The impact of paroxetine on macrophages has been explored in research, although a comprehensive understanding is still pending. This study aimed to research the potential of administering paroxetine to J774.2 macrophage cells to stimulate the release of GM-CSF, TNF-α, IL-12p40, and IL-6 cytokines. Additionally, we examined the mechanisms of action of paroxetine on the p38 signaling pathway, which is involved in cytokine production, and the PI3K pathway, which is an important mechanism in intracellular signaling. Our findings revealed that paroxetine induced an inflammatory response in macrophages by promoting cytokine synthesis in a non-lipopolysaccharide (LPS) environment. We observed that paroxetine triggered the inflammatory response through the PI3K signaling pathway while suppressing the p38 signaling pathway.

Role of lncRNAs in Remodeling of the Coronary Artery Plaques in Patients with Atherosclerosis.

INTRODUCTION: Cardiovascular diseases (CVDs) are the leading cause of death worldwide according to World Health Organization (WHO) data. Atherosclerosis is considered as a chronic inflammatory disease that develops in response to damage to the vascular intima-media layer in most cases. In recent years, epigenetic events have emerged as important players in the development and progression of CVDs. Since noncoding RNA (ncRNAs) are important regulators in the organization of the pathophysiological processes of the cardiovascular system, they have the potential to be used as therapeutic targets, diagnostic and prognostic biomarkers. In this study long noncoding RNA (lncRNA) and mRNA gene expression were compared between coronary atherosclerotic plaques (CAP) and the internal mammary artery (IMA)  which has the same genetic makeup and is exposed to the same environmental stress conditions with CAP in the same individual. METHODS: lncRNA and mRNA gene expressions were determined using the microarray in the samples. Microarray results were validated by RT-qPCR. Differentially expressed genes (DEGs; lncRNAs and mRNAs) were determined by GeneSpring (Ver 3.0) [p values < 0.05 and fold change (FC) > 2]. DAVID bioinformatics program was used for Gene Ontology (GO) annotation and enrichment analyses of statistically significant genes between CAP and IMA tissue. RESULTS AND CONCLUSIONS: In our study, 345 DEGs were found to be statistically significant (p < 0.05; FC > 2) between CAP and IMA. Of these, 65 were lncRNA and 280 were mRNA. Thirty-three lncRNAs were upregulated, while 32 lncRNAs were downregulated. Some of the important mRNAs are SPP1, CYP4B1, CHRDL1, MYOC, and ALKAL2, while some of the lncRNAs are LOC105377123, LINC01857, DIO3OS, LOC101928134, and KCNA3 between CAP and IMA tissue. We also identified genes that correlated with statistically significant lncRNAs. The results of this study are expected to be an important source of data in the development of new genetically based drugs to prevent atherosclerotic plaque. In addition, the data obtained may contribute to the explanation of the epigenetic mechanisms that play a role in the pathological basis of the process that protects the IMA from atherosclerosis.

Escitalopram's inflammatory effect on the mammalian macrophages and its intracellular mechanism of action.

The majority of patients with depression are treated with antidepressant drugs that are in the serotonin reuptake inhibitor (SSRI) group. Different studies have been conducted on the effect of treatment with antidepressants on the level of pro-inflammatory cytokines. There have been studies on the effects of escitalopram, an SSRI group antidepressant, on the pro-inflammatory cytokine levels both in vivo and in vitro. The results of these studies do not overlap and therefore the escitalopram's effect on the immune system should be studied in more depth. In this study, we aimed to examine, in detail, the cytokine production amount by escitalopram treatment of the J774.2 macrophage cells and its intracellular mechanism of action by examining the PI3K and p38 pathways. As a result of our study, we observed that Escitalopram caused a significant increase in TNF-α, IL-6, and GM-CSF levels in mammalian macrophage cells, but did not induce IL-12p40 production. We observed that the p38 and PI3K pathways play a role in inflammation in the presence of Escitalopram.

Anti-inflammatory activity of bupropion through immunomodulation of the macrophages.

Depression might manifest itself with a chronic inflammation in different tissues and organs independent of the central nervous system. Psoriasis, Crohn's disease, and fibromyalgia are among these disorders accompanying the depression. The treatment options for these conditions are a combination of the anti-depressants and anti-inflammatory agents. Bupropion has been widely utilized as an anti-depressant. It has been preferred among the patients with Crohn's disease and psoriasis due to its anti-inflammatory role, as well. In this study, we aimed to decipher its target in the immune system. Macrophages were activated in the presence of LPS and increasing concentrations of the bupropion. TNF-α, IL-6, GM-CSF, and IL-12p40 cytokines' production levels were measured by ELISA to compare it to the control groups. These cytokines have been associated with the aggressive inflammation in different tissues. Moreover, p38 and PI3K proteins' phosphorylated levels were measured to examine whether bupropion acts through these pathways or not. Our results suggest that bupropion had anti-inflammatory action on the activated macrophages and its mechanism of action was partially dependent on p38 but independent of PI3K pathways.

Conditioned media of mouse macrophages modulates neuronal dynamics in mouse hippocampal cells.

Many neurodegenerative diseases display both neuroinflammation and impaired neuron production in hippocampus. Although immunotherapeutic strategies indicate a promising avenue for combating neuroinflammation-induced diseases, directly targeting microglia, principle immune cells of CNS for such therapeutic purposes might be problematic due to invasive procedures. Instructing monocytes/macrophages from the periphery can be a less invasive and advantageous strategy compared to reaching microglia. But interplay between CNS neurons and macrophages even under normal conditions is poorly understood. To explore the experimental platform of how CNS derived neuronal cells respond to overall soluble factors of a non-CNS derived immune cell type, we introduced the conditioned media (CM) of unstimulated, and lipopolysaccharide stimulated RAW264.7 mouse macrophages to immortalized HT-22 mouse hippocampal cells during and after they were exposed to neuronal differentiation media. First, we recorded the cell viability of HT-22 cell study groups by using a real time cell analyzer. Then, we assessed the immunocytochemical expression of CR and CB proteins and mRNA levels of Ascl1, Bdnf, CB, Grn, Nrf2 and Rac1 genes via semi quantitative image analysis and q-RT-PCR among the different groups of HT-22 cells. Real time cell monitoring provided a solid physiological evidence regarding how various cell culture treatments affected the cell viability of HT-22 cells over time. Our further findings suggested that culturing HT-22 cells with unstimulated CM of macrophages markedly increased the immunocytochemical expression of CR and mRNA expression of Ascl1, Bdnf, CB and Grn genes, while the latter media resulted in decreases of those expressions. Overall, our results imply that HT-22 cells are meaningfully responsive to the secretome of RAW264.7 macrophages and using the interaction of macrophage with CNS derived neuronal cells is an instructive platform for deciphering the molecular mechanisms of cellular communication between immune system cells and neurons.

Immunostimulatory activity of fluoxetine in macrophages via regulation of the PI3K and P38 signaling pathways.

Fluoxetine is an antidepressant drug that is heavily preferred in the cure of depression, which is from the selective serotonin reuptake inhibitor (SSRI) group. There are many reports on the effect of fluoxetine on the immune system, and its effect on the macrophage cells has never been looked at before. We aimed to demonstrate the cytokine production potential of fluoxetine antidepressant, which is widely used in the clinic, in the J774.2 cell line and its effect on PI3K and P38 pathways. The use of fluoxetine alone in J774.2 macrophage cells showed immunostimulatory properties by inducing the production of tumor necrosis factor-α (TNF-α), interleukin (IL) IL-6, IL-12p40, and granulocyte-macrophage colony-stimulating factor (GM-CSF) cytokines. It showed anti-inflammatory properties by completely stopping the production of cytokines (IL-6, IL12p40, TNF-α, and GM-CSF) at all concentrations where LPS and fluoxetine were used together. While PI3K and P38 pathways were not effective in the immunostimulatory effect in the presence of the drug agent, we found that the PI3K and P38 pathways were influenced during their anti-inflammatory activity.

Apis mellifera anatoliaca Venom Exerted Anti-Inflammatory Activity on LPS-Stimulated Mammalian Macrophages by Reducing the Production of the Inflammatory Cytokines.

Extraction and characterization of natural products provide the opportunity to expand our arsenal of drug candidates against a wide range of diseases including cancer and inflammatory disorders. Previous studies have shown bee venom to have immense potential as an anti-inflammatory drug candidate. In this study, we focused on the venom of Apis mellifera anatoliaca and characterized its content by HPLC. An in vitro inflammation model based on lipopolysaccharide (LPS)-stimulated mammalian macrophages was utilized to examine the venom's anti-inflammatory potential. Additionally, its antiproliferative activity was evaluated in vitro against a human glioblastoma cell line. Based on the TNF, IL6, GMCSF, and IL12p40 pro-inflammatory cytokine production level in LPS-induced macrophages, venom-treated groups showed substantial decrease in the inflammatory action compared to untreated LPS-stimulated macrophages. When the cells were analyzed for viability, the venom did not have any cytotoxic effect on the macrophages at the concentration ranges that were utilized. Moreover, IC50 value of the venom was above 60 µg/mL on glioblastoma cancer cell line. These results suggest that the Apis mellifera anatoliaca venom does not have anticancer drug candidate potential, whereas it can efficiently be used against inflammatory and autoimmune disorders. To our knowledge, this is the first study to specifically examine the effect of anti-inflammatory activity of Apis mellifera anatoliaca venom on macrophages.

Non-canonical anti-cancer, anti-metastatic, anti-angiogenic and immunomodulatory PDT potentials of water soluble phthalocyanine derivatives with imidazole groups and their intracellular mechanism of action.

Cancer is currently a leading health issue globally. Chemotherapy is a prominent treatment method but due to undesired side effects t, there has been a need for novel less toxic approaches. Photodynamic therapy may be listed among the alternatives for efficient and potentially less detrimental applications of cancer therapy. Canonical photodynamic therapy (PDT) approach requires a light source with a specific wavelength of light, a non-toxic photosensitizer and molecular oxygen. PDT creates the desired effect by the photochemical reaction created through interaction of these components to create reactive oxygen species that will act on the cancer cells to enable anti-cancer activities. In our study we focus on non-canonical PDT application. In this approach we are not only aiming to eliminate cancer cells in the environment but also test the anti-metastatic, anti-angiogenic and possible immunomodulatory activities of the novel photosensitizers. Moreover, in our approach, we studied the intracellular pathways that are crucial for carcinogenesis, cell cycle, apoptosis, angiogenesis, metastasis and immune function to decipher the mechanism of the action for each compound. Reactive oxygen species based explanation was not valid in our study, hence it brings out a non canonical approach to PDT applications. Our results suggests that Phthalocyanine derivatives with imidazole groups can be effectively used against lung, colon, breast and prostate cancer while differentially effecting metastasis, angiogenesis, cell cycle, apoptosis and immune system cells' activities. Based on the results, PDT application of these phthalocyanine derivatives can be an effective treatment option to replace chemotherapy to minimize the potential side effects.

Photodynamic anti-inflammatory activity of azulene derivatives on mammalian macrophages and their intracellular mechanism of action.

Azulene derivatives have been studied previously as photodynamic therapy agents. They have anti-cancer, anti-microbial and anti-inflammatory activities. Together with their photodynamic activity they enable more control on their activation which aims to decrease possible side effects that have been encountered with their constitutively active drug counterparts. In our current study we focused on photodynamic anti-inflammatory activities of two azulene derivatives whose synthesis methods were described before. We found that when mammalian macrophages J774.2 cells were incubated with these two derivatives in the presence of LPS in dark conditions, these molecules had anti-inflammatory activity at their highest concentrations based on ELISA results on the pro-inflammatory cytokine levels. After light application, both derivatives exerted strong anti-inflammatory activities by substantially decreasing the TNF, IL6, GMCSF and IL12p40 cytokine production levels. When the intracellular mechanism of action for both derivatives was tested, only one of them acted through p38 and PI3K pathways whereas the other derivative did not affect either of these pathways. Our results suggest that these two azulene derivatives can be utilized as photodynamic anti-inflammatory drug candidates.

Synthesis of novel immunomodulatory 1,4-disubstituted bis-1,2,3-triazoles by using click chemistry and their intracellular mechanism of action.

In this study, six new 1,4-disubstituted bis-1,2,3-triazole compounds, N,N'-(1,2-phenylene)bis(2-(4-R-1H-1,2,3-triazol-1-yl)acetamide), were synthesized with high yield (88-96 %) by using click chemistry and their molecular structures were characterized by using NMR, FT-IR, HRMS and elemental analysis techniques. Previous studies suggest anti-inflammatory and analgesic activities for different 1,2,3-triazole derivatives and in the light of those studies we aimed to examine these novel derivatives immunomodulatory activities on the mammalian macrophages. Pro-inflammatory cytokines (TNF, IL6, GMCSF and IL12p40) secretion levels were tested in the presence of bis-1,2,3-triazole compounds when the macrophages were activated with LPS. These new derivatives were able to suppress the production of these cytokines at different levels. Intracellular phophorylated PI3K protein levels were measured due to its prominent role in inflammatory reactions. Our flow cytometry analysis results suggested that some of these compounds were partially effective through PI3K pathway. In different inflammatory and autoimmune disease settings these novel 1,2,3-triazole derivatives can be utilized as non-steroid based anti-inflammatory drug candidates.

Water-Based Synthesis of Copper Chalcogenide Structures and Their Photodynamic Immunomodulatory Activities on Mammalian Macrophages.

Generation of novel and versatile immunomodulatory agents that could suppress excessive inflammation has been crucial to fight against chronic inflammatory and autoimmune disorders. Immunomodulatory agents regulate the function of immune system cells to manage their activities. Current therapy regimens for the inflammatory and autoimmune disorders rely on immunomodulatory drug molecules but they are also associated with unwanted and severe side effects. In order to prevent the side effects associated with drug molecules, the field should generate novel immunomodulatory drug candidates and further test them. Moreover, the generation of photodynamic immunomodulatory molecules would also decrease possible side effects. Photodynamic activation enables specific and localized activation of the active ingredients upon exposure to a certain wavelength of light. In our study, we generated copper-based chalcogenide structures in gel and nanoparticle form by using a water-based method so that they are more biocompatible.After their chemical characterization, they were tested on mammalian macrophages in vitro. Our results suggest that these molecules were anti-inflammatory in dark conditions and their anti-inflammatory potentials significantly increased upon xenon light treatment. We are presenting novel photodynamic immunomodulatory agents that can be used to suppress excessive inflammation in disease conditions that have been associated with excessive inflammation.

A Review on the Design, Synthesis, and Structure-activity Relationships of Benzothiazole Derivatives against Hypoxic Tumors.

There has been a growing body of studies on benzothiazoles and benzothiazole derivatives as strong and effective anti-tumor agents against lung, liver, pancreas, breast, and brain tumors. Due to the highly proliferative nature of the tumor cells, the oxygen levels get lower than that of normal tissues in the tumor microenvironment. This situation is called hypoxia and has been associated with increased ability for carcinogenesis. For the drug design and development strategies, the hypoxic nature of the tumor tissues has been exploited more aggressively. Hypoxia itself acts as a signal initiating system to activate the pathways that eventually lead to the spread of the tumor cells into the different tissues, increases the rate of DNA damage, and eventually ends up with more mutation levels that may increase the drug resistance. As one of the major mediators of hypoxic response, hypoxia-inducible factors (HIFs) have been shown to activate angiogenesis, metastasis, apoptosis resistance, and many other protumorigenic responses in cancer development. In the current review, we will be discussing the design, synthesis, and structureactivity relationships of benzothiazole derivatives against hypoxic tumors such as lung, liver, pancreas, breast, and brain as potential anti-cancer drug candidates. The focus points of the study will be the biology behind carcinogenesis and how hypoxia contributes to the process, recent studies on benzothiazole and its derivatives as anti-cancer agents against hypoxic cancers, conclusions, and future perspectives. We believe that this review will be useful for researchers in the field of drug design during their studies to generate novel benzothiazole-containing hybrids against hypoxic tumors with higher efficacies.

Novel benzoylthiourea derivatives had differential anti-inflammatory photodynamic therapy potentials on in vitro stimulated mammalian macrophages.

Novel benzoylthioureas, N-((5-chloropyridin-2yl)carbamothioyl)benzamide, (HL1), N-((2-chloropyridin-3yl)carbamothioyl)benzamide, (HL2), N-((5-bromopyridin-2yl)carbamothioyl)benzamide, (HL3) and N-(Naphthalene-1-yl(phenyl)carbamothioyl)benzamide, (HL4), were synthesized. Their characterizations were made by FT-IR,1H NMR and 13C NMR spectrophotometric analysis. Single crystal X-ray diffraction measurements were conducted to determine the crystal structure of HL1 and HL4.  The HL1 crystallization conditions are: in the monoclinic crystal system with P21/c space group, Z = 2, a = 8.118(2) Å, b = 12.056(3) Å, c = 13.753(4) Å. HL4crystallization conditions are: in the orthorhombic crystal system with Pbca space group, Z = 8, a = 19.597(9) Å, b = 8.270(4) Å, c = 24.299(11) Å. Investigation of photodynamic and antiinflamatory effects of these compounds revealed that they are potent adducts. Using these derivatives, mammalian macrophages were stimulated with LPS to test their anti-inflammatory activity. Based on pro-inflammatory cytokine production levels, the photodynamic anti-inflammatory activity of these adducts were found to differ. Our results showedthat benzoylthioureas can be used as potential photodynamic therapy agents to suppress the excessive inflammatory reactions encountered in autoimmune and inflammatory disorders.

Differential anti-inflammatory properties of chitosan-based cryogel scaffolds depending on chitosan/gelatin ratio.

Chitosan/gelatine-based materials have been widely used as biocompatible scaffolds in the tissue engineering field. Chitosan suppresses the inflammatory activities of macrophages whereas gelatine induces inflammatory cytokine production by these cells. Cryogel form of the scaffolds created an effect that was mostly dominated by chitosan activity. Since independent of chitosan to gelatine ratio, the cryogels eliminated the inflammatory cytokine production by the activated macrophages. This will enable suppression of inflammatory reactions by macrophages during implant procedure while enabling a nest of the matrix for the macrophages to reside. Determining the immunomodulatory effect of these materials during the decay is crucial to assess their biocompatibility and safety. Our results suggest that when the chitosan ratio was higher than that of gelatine the materials had anti-inflammatory activity in their powder forms based on TNFα production levels by LPS activated macrophages, whereas higher gelatine to chitosan ratio eliminated this effect. To our knowledge, this is the first study to assess the powder vs. gel forms of the chitosan/gelatine-based materials for their immunomodulatory potentials as well as how the ratio of chitosan to gelatine might affect these materials immunomodulatory effects on the activated macrophages.HIGHLIGHTSChitosan/gelatin composite cryogels have anti-inflammatory activities.Different ratios of chitosan to gelatin content altered the immunomodulatory activities.They can be safely and effectively used as implant materials for tissue engineering applications.They will also reduce the use of anti-inflammatory drugs during implantation.

Antiproliferative activity of Tamoxifen, Vitamin D3 and their concomitant treatment.

Breast cancer stands out as the most common cancer type among women throughout the world. Especially for the estrogen receptor alpha (ER α +) positive breast cancer cells Tamoxifen has been widely used as an anti-cancer agent. Tamoxifen's mechanism of action is through ER. It binds to the receptor and leads to a conformational change which eventually prevents cancer cells proliferation and survival. In our current study, we aimed to investigate the combination of Tamoxifen with Vitamin D3 to test whether this combination will enhance the anti-cancer effect of Tamoxifen on breast cancer cells in vitro. Vitamin D3 has sterol structure and this property enables it to act similar to hormones. Vitamin D Receptor (VDR) has been commonly found in different types of cancer cells including but not limited to breast and prostate cancer cells. Through this receptor Vitamin D3 acts as an anti-proliferative agent. We examined the proliferation rate, apoptosis and necrosis levels as well as cell cycle progression in MCF-7 breast cancer cell line in the presence of Vitamin D3 and Tamoxifen to compare the changes with the Tamoxifen treated group. Our results suggest that Tamoxifen was a more potent anti-cancer agent than Vitamin D3 or its combination with Vitamin D3 based on cell cycle arrest, apoptosis and cell proliferation levels. This effect in the apoptosis rate and cell cycle stage of the MCF-7 cells were in line with the changes in gene expression profile of P53, BAX and BCL-2. Our results suggest that Tamoxifen by itself is adequate enough and more potent than Vitamin D3 or its combination with Vitamin D3 as anti-cancer agent for the breast cancer cells in vitro.