Synthesis and Characterization of Piano-Stool Ruthenium(II)-Arene Complexes of Isatin Schiff Bases: Cytotoxicity and DNA Intercalation.

A series of aryl-isatin Schiff base derivatives (3a-d) and their piano-stool ruthenium complexes (4a-d) were synthesized and characterized via 1H and 13C NMR and Fourier transform infrared (FTIR) spectroscopy. In addition, the purity of all of the compounds (3a-c and 4a-d) was determined via elemental analysis. Complex 4d was analyzed using X-ray crystallography. An in vitro antiproliferative study of the compounds (3a-c and 4a-d) against human hepatocellular carcinoma (HEPG2), human breast cancer (MCF-7), human prostate cancer (PC-3), and human embryonic kidney (HEK-293) cells exhibited their considerable antiproliferative activity. 4d exhibited effective cytotoxicity against HEPG2 and MCF-7. It displayed higher cytotoxicity than the reference metallo-drug cisplatin. Moreover, the stability of 4d was studied via 1H NMR spectroscopy, and the binding model between 4d and DNA was investigated via ultraviolet-visible spectroscopy. The lipophilicity of the synthesized complexes was determined using an extraction method.

A New Approach to Melanoma Treatment: microRNAs.

Although immunotherapy and targeted therapy have radically changed melanoma treatment, the development of resistance and reduction of patient responses are still significant problems. Small molecule inhibitors are needed to overcome this situation, and biomarkers that can estimate whether patients will reply to existing treatments need to be developed. miRNAs are involved in diverse processes such as tumor development, tumor progression, metastasis, and invasion. While some miRNAs act as tumor suppressors, others may be oncogenic. miRNAs also contribute to the processes involved in drug resistance. There is increasing evidence demonstrating the possible effect of miRNAs on the diagnosis and treatment markers of melanoma. The manuscript focuses on the current challenges in melanoma treatment, highlighting issues such as the development of resistance and reduced patient responses despite the revolutionary advancements in targeted therapy and immunotherapy. It underscores the need for small molecule inhibitors and the creation of biomarkers for predicting patient responses to current treatments. The role of miRNAs in processes such as tumor development, metastasis, and invasion has been highlighted. While certain miRNAs function as tumor suppressors, others may exhibit oncogenic properties. Furthermore, increasing evidence is presented demonstrating the potential significance of miRNAs as markers for the symptom and identification of melanoma. These findings indicate a promising avenue for future research and clinical applications. In summary, the article effectively communicates key insights, making it a valuable resource for those interested in melanoma research and treatment.

Chitosan in cancer therapy: a dual role as a therapeutic agent and drug delivery system.

Although chemotherapy is still the most preferred treatment for cancer, most chemotherapeutic agents target both cancer cells and healthy cells and cause serious side effects due to high toxicity. Improved drug delivery systems (DDSs), which enhance the efficacy of current chemotherapeutic drugs while reducing their toxicity, offer potential solutions to these challenges. Chitosan (CS) and its derivatives are biopolymers with biodegradable, biocompatible, and low-toxicity properties, and their structure allows for convenient chemical and mechanical modifications. In its role as a therapeutic agent, CS can impede the proliferation of tumor cells through the inhibition of angiogenesis and metastasis, as well as by triggering apoptosis. CS and its derivatives are also frequently preferred as DDSs due to their properties such as high drug-carrying capacity, polycationic structure, long-term circulation, and direct targeting of cancer cells. Various therapeutic agents linked to CS and its derivatives demonstrate potent anticancer effects with advantages such as reduced side effects compared to the original drugs, owing to factors like targeted distribution within cancer tissues and sustained release. This review emphasizes the utilization of CS and its derivatives, both as therapeutic agents and as carriers for established chemotherapeutic drugs.

In vitro cytotoxicity of magnetic-fluorescent bioactive glasses on SaOS-2, MC3T3-E1, BJ fibroblast cells, their hemolytic activity, and sorafenib release behavior.

In the study, the fabrication of superparamagnetic-fluorescent bioactive glasses in the form of the particle, nanofiber, and 3D scaffolds was performed by including maghemite (γ-Fe2O3) nanoparticles and photoluminescent rare earth element ions (Eu3+, Gd3+, and Yb3+) using sol-gel, electrospinning, and robocasting techniques, respectively. The in vitro cytotoxicity of the magnetic-fluorescent bioactive glasses on osteosarcoma SaOS-2, pre-osteoblast MC3T3-E1, and BJ fibroblast cells, as well as their hemolytic activity and sorafenib tosylate loading and release behavior, were investigated. The cytotoxicity of the bioactive glass samples was tested using the MTT assay. Additionally, the alkaline phosphatase activity of the studied glasses was examined as a function of time. The mineralization behavior of the pre-osteoblast cell-seeded glass samples was analyzed using Alizarin red S staining. Results revealed that the in vitro cytotoxicity of the studied bioactive glasses in the form of particles and nanofibers depended on the sample concentration, whereas in the case of the 3D scaffolds, no cytotoxic response was observed on the osteosarcoma, pre-osteoblast, and fibroblast cells. Similarly, particle and nanofiber-based glass samples induced dose-dependent hemolysis on red blood cells. Drug loading rates were much lower for the 3D scaffolds compared to the particle and nanofiber-based samples. Drug release rates ranged from 25 % to 90 %, depending on the bioactive glass morphology and the pH of the release medium. It was concluded that the studied bioactive glasses have the potential to be used in tissue engineering applications and cancer therapy.

Salivary inflammatory burden in pre- and postmenopausal women: Associations with body mass index, patient-reported health, serum cytokines, and periodontal parameters.

BACKGROUND: The decline of estrogen levels during menopause impacts weight, mood, and overall health, both orally and systemically. This study assessed salivary levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), IL-10, and IL-7 in postmenopausal (PMW) and regularly menstruating premenopausal (RMPW) women, while considering serum cytokine levels, body mass index (BMI), periodontal health, and self-reported physical and emotional well-being. METHODS: In this study, 75 PMW and 71 RMPW were included. Clinical and periodontal parameters were evaluated, and perceived health was assessed with the Women's Health Questionnaire (WHQ). Cytokine levels in both saliva and serum were quantified by enzyme-linked immunosorbent assay (ELISA). Covariate evaluations of salivary cytokines were conducted using hierarchical linear regression modeling. RESULTS: Cytokines were detectable in saliva from 71 PMW and 67 RMPW. In the initial unadjusted model, IL-7, IL-10, and TNF-α exibited significant differences between RMPW and PMW. However, these differences became non-significant (p > 0.05) in the final model after adjusting for age, which implies a negligible effect of the investigated covariates on salivary cytokine levels when age was considered. Lower levels of IL-6 in PMW, which initially showed no significant difference, became borderline (p = 0.054) in the final model after adjusting for age. CONCLUSIONS: After adjusting for multiple factors, no significant difference was found in the salivary levels of the investigated cytokines between RMPW and PMW. Factors such as BMI, perceived health, serum cytokine levels, and periodontal parameters seem to minimally influence these levels in PMW. However, age may be a stronger confounding factor.

Effects of acute exposure to environmentally realistic tebuconazole concentrations on stress responses of kidney and digestive gland of Lymnaea stagnalis.

This study aimed to investigate the effects of 24 and 72 h exposure to environmentally relevant concentrations of tebuconazole (TEB) (10, 100 and 500 µg/L) fungicide on the freshwater snail Lymnaea stagnalis. The focus was induction of oxidative stress, alteration of gene expressions and histopathological changes in the kidney and digestive gland. TEB treatment induced a time- and concentration-dependent increase in intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) levels, while the total antioxidant capacity (TAC) was decreased. The activities of glutathione peroxidase (GPx), glutathione reductase (GR), and catalase (CAT) also increased in a time- and concentration-dependent manner in both tissues. TEB exposure significantly increased the mRNA levels of CAT, GPx, GR, heat shock proteins HSP40 and HSP70. Histological analysis revealed nephrocyte degeneration and disrupted digestive cells. The study concludes that acute exposure to TEB induces oxidative stress, alters antioxidant defense mechanisms, and leads to histopathological changes in L. stagnalis.

Bioevaluation of Spiro N-Propargylic ß-Enaminones as Anti-Breast Cancer Agents: In Vitro and Molecular Docking Studies.

The study aimed to investigate the in vitro inhibitory activities of spiro N-propargylic ß-enaminones, SPEs 1-31, against BCa cells, to perform in silico molecular docking studies to understand the nature of the interaction between the compounds and the ERα, PR, EGFR, and Her2, and to determine the ADMET and drug-likeness properties. Cytotoxic activity was investigated via MTT assay. DNA fragmentation was evaluated via ELISA assay. Cell cycle distributions were investigated by flow cytometry. Expression levels of Bcl-2, Bax, p21 and Cyclin D1 were measured by qRT-PCR and western blot analysis. Molecular docking was done using Autodock/vina software. ADMET analysis was calculated using the ADMETlab 2.0 tool. SPEs 1, 22, and 28 showed selective cytotoxic activity against all BCa cells with SI values >2. SPEs induced apoptosis and caused significant changes in Bcl-2 and Bax levels. The cell cycle was arrested at the S phase and levels of p21 and Cyclin D1 were induced in all BCa cells. Molecular docking analysis revealed that SPE1, SPE22, and SPE28 showed high binding affinities with ERα, PR, EGFR, and Her2. ADMET analysis revealed that SPEs are drug-like compounds as they obey the five rules of Lipinsky and are not toxic. Therefore, these potential anticancer compounds should be further validated by in vivo studies for their appropriate function in human health with a safety profile, and a comprehensive drug interaction study should be performed.

Effects of leptin on the viability of human ovarian cancer cells and changes in cytokine expression levels.

Background: Obesity is associated with increased mortality among ovarian cancer and is a poor prognostic factor. There are significant links between the leptin hormone, a product of the obesity gene, and the development of ovarian cancer. Leptin is a vital hormone-like cytokine secreted from adipose tissue and is mainly involved in the maintenance of energy homeostasis. It regulates several intracellular signaling pathways and also interacts with various hormones and energy regulators. It acts as a growth factor by stimulating cell proliferation and differentiation and in this way contributes to cancer cell development. The aim of the study was to investigate the effects of leptin on human ovarian cancer cells. Methods: In this study, the effects of increasing the concentration of leptin were investigated on the cell viability of OVCAR-3 and MDAH-2774 ovarian cancer lines by MTT assay. Moreover, to elucidate the molecular mechanisms of leptin in ovarian cancer cells, changes in the expression levels of 80 cytokines were evaluated after leptin treatment via a human cytokine antibody array. Results: Leptin increases the proliferation of both ovarian cancer cell lines. IL-1 level was increased in OVCAR-3 cells and TGF-ß level was increased in MDAH-2774 cells after leptin treatment. A decrease in IL-2, MCP-2/CCL8 and MCP-3/CCL7 levels was detected in both ovarian cancer cell lines with leptin administration. An increase in IL-3 and IL-10 expressions, insulin-like growth factor binding proteins (IGFBP) IGFBP-1, IGFBP-2 and IGFBP-3 levels were detected in both ovarian cancer cell lines with leptin administration. In conclusion; leptin has a proliferative effect on human ovarian cancer cell lines and affects different cytokines in different types of ovarian cancer cells.

N-Propargylic ß-enaminones in breast cancer cells: Cytotoxicity, apoptosis, and cell cycle analyses.

Breast cancer is one of the most common cancers worldwide and the discovery of new cytotoxic agents is needed. Enaminones are regarded to be a significant structural motif that is found in a variety of pharmacologically active compounds however the number of studies investigating the anticancer activities of N-propargylic ß-enaminones (NPEs) is limited. Herein we investigated the potential cytotoxic and apoptotic effects of 23 different NPEs (1-23) on human breast cancer cells. Cytotoxicity was evaluated via MTT assay. Apoptotic cell death and cell cycle distributions were investigated by flow cytometry. CM-H2DCFDA dye was used to evaluate cellular ROS levels. Expression levels of Bcl-2, Bax, p21, and Cyclin D1 were measured by quantitative real-time PCR. ADME properties were calculated using the ADMET 2.0 tool. NPEs 4, 9, 16, and 21 showed selective cytotoxic activity against breast cancer cells with SI values >2. NPEs induced apoptosis and caused significant changes in Bcl-2 and Bax mRNA levels. The cell cycle was arrested at the G0/G1 phase and levels of p21 and Cyclin D1 were upregulated in both breast cancer cells. ROS levels were significantly increased by NPEs, suggesting that the cytotoxic and apoptotic effects of NPEs were mediated by ROS. ADME analysis revealed that NPEs showed favorable distributions in both breast cancer cell lines, meaning good lipophilicity values, low unfractionated values, and high bioavailability. Therefore, these potential anticancer compounds should be further validated by in vivo studies for their appropriate function in human health with a safety profile, and a comprehensive drug interaction study should be performed.

Design and synthesis of benzimidazole derivatives as apoptosis-inducing agents by targeting Bcl-2 protein.

Bcl-2, an anti-apoptotic protein, is a well-known and appealing cancer therapy target. Novel series of benzimidazole derivatives were synthesized and tested for their activity as Bcl-2 inhibitors on T98G glioblastoma, PC3 prostate, MCF-7 breast, and H69AR lung cancer cells. MTT assay was used to evaluate the cytotoxic effect. PI Annexin V Apoptosis Detection Kit was used to detect apoptosis. Expression levels of the Bcl-2 protein were examined by the Western blot analysis and qRT-PCR. All synthesized benzimidazole derivatives exhibited a cytotoxic effect on cancer cells with IC50 values in the range of 25.2-88.2 µg/mL. Among all derivatives, compounds C1 and D1 demonstrated a higher cytotoxic effect on cancer cells with IC50 values < 50 µg/mL, while a lower cytotoxic effect against human embryonic kidney cells with IC50 values of > 100 µg/mL. C1 and D1 caused a significant increase in the percentage of apoptotic cells in all types of cancer cell cells and both Bcl-2 mRNA and protein levels were significantly reduced. These results suggest that the novel benzimidazole derivatives may be candidates for apoptosis-inducing agents in cancer treatment by targeting anti-Bcl-2 proteins in cancer cells.

Trabectedin (ET-743) in prostate cancer: Endoplasmic reticulum stress-induced apoptotic effect.

Trabectedin is a chemotherapy agent originating from a tunicate, Ecteinascidia turbinata. In this study, DNA-independent action mechanisms of trabectedin are investigated in prostate cancer (PCa) cells. Cell viability was assessed via XTT assay. Apoptosis was evaluated via flow cytometry. Tetramethylrodamine ethyl ester (TMRE) dye was utilized to determine mitochondrial membrane potential (MMP). Cell cycle distribution was investigated via flow cytometric analysis. Reactive oxygen species (ROS) were monitored using fluorescence CM-H2DCFDA dye. Changes in CHOP, p-eIF2α, GRP78 and p-PERK which are endoplasmic reticulum (ER) stress-involved proteins were investigated via western blot. Trabectedin induced cytotoxicity and cell cycle arrest at the G2/M phase. Trabectedin decreased MMP via ROS generation in PCa cells. ER stress-related proteins CHOP, p-eIF2α, GRP78 and p-PERK were also elevated by trabectedin treatment indicating the induction of ER stress-induced apoptosis. The results of this study show that trabectedin may be an effective chemotherapeutic for PCa.

Synthesis of silver nanoparticles using Alpinia officinarum rhizome extract induces apoptosis through down-regulating Bcl-2 in human cancer cells.

In this study, silver nanoparticles were synthesized using Alpinia officinarum rhizome extract via an eco-friendly green synthesis method. The silver nanoparticles (AO-AgNPs) were characterized by UV-Vis spectrometry, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and dynamic light scattering. Further, the cytotoxic and apoptotic effects of AO-AgNPs were investigated in human cancer cells with different tissue origins via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and flow cytometric analyses, respectively. The expression levels of anti-apoptotic Bcl-2 protein were evaluated via a real-time polymerase chain reaction. The synthesized AO-AgNPs induced a significant cytotoxic effect in all tested cancer cells but not in normal cells. AO-AgNPs induced the percentage of apoptotic cells and reduced the levels of anti-apoptotic Bcl-2 mRNA levels in cancer cells. These results demonstrate the potential application of AO-AgNPs in cancer treatment.

Endoplasmic Reticulum Stress-Induced Apoptotic Effects of Novel 1-Pyrroline (3,4-Dihydro-2H-pyrrole) Derivatives on Breast Cancer Cells.

Heterocyclic compounds have emerged as promising and appealing scaffolds for developing effective antitumor agents. Here, the effects of synthesized 24 different 1-pyrroline derivatives (PDs) containing substituted aryl sulfide moiety were investigated on human breast cancer cell lines. The viability of cells was assessed via MTT assay. Reactive oxygen species (ROS) generation was analyzed via fluorescent dye CM-H2DCFDA. Apoptotic cells were determined via flow cytometry. Endoplasmic reticulum (ER) stress-associated protein levels were analyzed via western blot analysis. Four of the PDs (PD-12, -14, -16 and -17) had great cytotoxic selectivity against breast cancer cells. Apoptotic cell death was induced by PDs via the generation of ROS. PDs significantly increased the GRP78, p-PEAK, p-eIF2α, and CHOP protein levels indicating ER stress in breast cancer cells. These results imply that newly synthesized PDs may be potential anticancer agents as they selectively inhibit breast cancer cells.

mRNA as a Therapeutics: Understanding mRNA Vaccines.

Vaccination is one of the important approaches in the prevention and control of diseases. Although the capacity to present antigens other than the disease-specific antigen in the traditional vaccine composition provides a potential benefit by increasing its protective efficacy, many components that are not needed for the related disease are also transferred. These components can reduce vaccine activity by lowering immunity against protective antigens. The reasons such as the low effectiveness of traditional vaccines and the high cost of production and time-consuming reasons show that it is necessary to develop a new vaccine method for our world, which is struggling with epidemics almost every year. Among nucleic acids, mRNA has many advantages, such as genomic integration, induction of anti-DNA autoantibodies, and immune tolerance induced by long-term antigen expression. mRNA vaccines have become a therapeutic target for reasons such as efficacy, safety, fast and non-expensive production. The fact that mRNA triggers both humoral and cellular immunity and goes only to the cytoplasm, not to the nucleus, makes it highly efficient. The mRNA must cross the lipid bilayer barrier and entry to the cytoplasm where it is translated into protein. There are two main ways of mRNA vaccine delivery for this: ex vivo loading of mRNA into dendritic cells (DCs) and direct injection of mRNA with or without a carrier. Studies continue to understand which delivery system is therapeutically more efficient. Preclinical and clinical trials showed that mRNA vaccines trigger a long-lasting and safe immune response.

Drug Delivery Systems for Cancer Treatment: A Review of Marine-derived Polysaccharides.

Cancer is a disease characterized by uncontrolled cell proliferation and the spread of cells to other tissues and remains one of the worldwide problems waiting to be solved. There are various treatment strategies for cancer, such as chemotherapy, surgery, radiotherapy, and immunotherapy, although it varies according to its type and stage. Many chemotherapeutic agents have limited clinical use due to lack of efficacy, off-target toxicity, metabolic instability, or poor pharmacokinetics. One possible solution to this high rate of clinical failure is to design drug delivery systems that deliver drugs in a controlled and specific manner and are not toxic to normal cells. Marine systems contain biodiversity, including components and materials that can be used in biomedical applications and therapy. Biomaterials such as chitin, chitosan, alginate, carrageenan, fucoidan, hyaluronan, agarose, and ulvan obtained from marine organisms have found use in DDSs today. These polysaccharides are biocompatible, non-toxic, biodegradable, and cost-effective, making them ideal raw materials for increasingly complex DDSs with a potentially regulated release. In this review, the contributions of polysaccharides from the marine environment to the development of anticancer drugs in DDSs will be discussed.

Liquidambar orientalis Mill. gum extract induces autophagy via PI3K/Akt/mTOR signaling pathway in prostate cancer cells.

Liquidambar orientalis Mill (LOM), is an endemic species having a local distribution in the southwestern coastal district of Turkey. Styrax liquidus gum (SLG), is a gum-like resinous which exudates in response to injury of the trunk of LOM. The aim of the study was to investigate the cytotoxic effects and the molecular mechanisms of the ethanolic SLG extract in human prostate cancer cells. GC-MS analysis was performed to identify the volatile compound composition. Cytotoxicity was determined by XTT analysis. Apoptosis and necrosis were evaluated via ELISA assay. Autophagic cell death was detected via monodansylcadaverine (MDC) staining and by measuring the levels of LC3I and LC3II. The protein levels of p-PI3K, p-Akt and p-mTOR were evaluated by western blot analysis. In the present study, it is shown that the SLG extract containing a considerable amount of ravidomycin derivate induced autophagic cell death in prostate cancer cells via inhibiting the PI3K/Akt/mTOR pathway.

Preparation, Characterization, and Drug Delivery of Hexagonal Boron Nitride-Borate Bioactive Glass Biomimetic Scaffolds for Bone Tissue Engineering.

In this study, biomimetic borate-based bioactive glass scaffolds containing hexagonal boron nitride hBN nanoparticles (0.1, 0.2, 0.5, 1, and 2% by weight) were manufactured with the polymer foam replication technique to be used in hard tissue engineering and drug delivery applications. To create three-dimensional cylindrical-shaped scaffolds, polyurethane foams were used as templates and covered using a suspension of glass and hBN powder mixture. Then, a heat treatment was applied at 570 °C in an air atmosphere to remove the polymer foam from the structure and to sinter the glass structures. The structural, morphological, and mechanical properties of the fabricated composites were examined in detail. The in vitro bioactivity of the prepared composites was tested in simulated body fluid, and the release behavior of gentamicin sulfate and 5-fluorouracil from glass scaffolds were analyzed separately as a function of time. The cytotoxicity was investigated using osteoblastic MC3T3-E1 cells. The findings indicated that the hBN nanoparticles, up to a certain concentration in the glass matrix, improved the mechanical strength of the glass scaffolds, which mimic the cancellous bone. Additionally, the inclusion of hBN nanoparticles enhanced the in vitro hydroxyapatite-forming ability of bioactive glass composites. The presence of hBN nanoparticles accelerated the drug release rates of the system. It was concluded that bioactive glass/hBN composite scaffolds mimicking native bone tissue could be used for bone tissue repair and regeneration applications.

4-Propargyl-substituted 1H-pyrroles induce apoptosis and autophagy via extracellular signal-regulated signaling pathway in breast cancer.

Novel pyrrole derivatives (PDs) with propargyl units (1-7) were investigated for their anticancer activity on breast cancer cells. The MTT assay was used to assess the cell viability. Morphological changes in human breast cancer cells were visualized under a phase-contrast microscope. Apoptosis and autophagy were detected using the DNA fragmentation assay and staining by autophagic vacuoles, respectively. The levels of apoptosis- and autophagy-related proteins such as cytochrome c, Bcl-2, LC3-I/II were investigated by Western blot analysis. The effect of PDs on the ERK1/2 signaling pathway was investigated using specific inhibitors. All the tested PDs were found to be active in the range of 36.7 ± 0.2 to 459.7 ± 4.2 µM. Compounds 3 and 4 showed cytotoxic activity in breast cancer cells, but were found to be safer with lower cytotoxicity on human nontumorigenic epithelial breast cells. Compound 4 induced apoptosis, whereas compound 3 induced autophagy. Both compounds inhibited the ERK signaling pathway in breast cancer cells. The present study revealed that both synthesized PDs induced different programmed cell death types by inhibiting the ERK signaling pathway in two genotypically different breast cancer cells. Therefore, novel PDs might be promising anticancer agents for breast cancer therapy and further structural modifications of PDs may yield promising anticancer agents.

Does peri-implant bone loss affect the LL-37 and proteinase 3 levels in peri-implant sulcus fluid?

BACKGROUND: Inactive human cathelicidin antimicrobial peptide is present in neutrophils, and proteinase 3 activates this peptide by producing active LL-37 peptide. LL-37 acts as a defensive peptide in the oral tissues. In the present study, the aim was to evaluate LL-37 and proteinase 3 levels in peri-implant sulcus fluid (PISF) in implants with and without peri-implantitis. METHODS: Patients who simultaneously had dental implants with peri-implantitis and without peri-implantitis were included in the study. Forty-four samples with peri-implantitis and 34 samples without peri-implantitis from 16 patients were obtained. Intraoral evaluations such as pocket depth, modified sulcus bleeding index, and modified plaque index were noted. Enzyme-linked immunosorbent assay was used for the evaluation of PISF LL-37 and proteinase 3 levels. RESULTS: PISF volume was significantly increased in the implants with peri-implantitis than those without peri-implantitis (p < 0.05). No difference was present between PISF LL-37 and proteinase 3 total amounts between the implants with and without peri-implantitis (p > 0.05). Pocket depths and PISF LL-37 and proteinase 3 levels were not correlated in the groups (p > 0.05). CONCLUSIONS: PISF volume might be increased in response to peri-implant bone destruction. However, peri-implant tissue destruction caused by peri-implantitis does not seem to affect PISF LL-37 and proteinase 3 levels.

Novel benzimidazole derivatives: Synthesis, in vitro cytotoxicity, apoptosis and cell cycle studies.

The aim of the study was to synthesize a new series of benzimidazole derivatives and to investigate the underlying molecular mechanisms of the potential cell cycle inhibition and apoptotic effects against a panel of selected human cancer cell lines along with HEK-293 human embryonic kidney cells. MTT assay was used to evaluate cytotoxic effects. Muse™ Cell Analyzer was used to assess cell cycle progression. Annexin-V/PI staining assay was used for detecting apoptosis. All the synthesized compounds showed a significant cytotoxic effect against cancer cells with the IC50 values between 9.2 and 166.1 µg/mL. Among the tested derivatives, compound 5 showed significant cytotoxic activity against MCF-7, DU-145 and H69AR cancer cells with the IC50 values of 17.8 ± 0.24, 10.2 ± 1.4 and 49.9 ± 0.22 µg/mL respectively. The compounds 5 was also tested on HEK-293 human embryonic kidney cells and found to be safer with lesser cytotoxicity. The results revealed that compound 5 significantly increased cell population in the G2/M-phase which is modulated by a p53 independent mechanism. Compound 5 caused an increase in the percentage of late apoptotic cells in all tested cancer cells in a concentration-dependent manner. Among all synthesized derivatives, compound 5 the bromo-derivative, showed the highest cytotoxic potential, induced G2/M cell cycle arrest and apoptotic cell death in genotypically different human cancer cells. These results suggest that compound 5 might be a promising agent for cancer therapy and further structural modifications of benzimidazole derivatives may create promising anticancer agents.