Search

Drug-induced cardiotoxicity is a life-threatening side effect of doxorubicin (DOX) treatment that impacts patient prognosis and survival. In the majority of cases, the acute clinical form often remains asymptomatic, with few patients presenting rather nonspecific electrocardiographic abnormalities. While chronic toxicity has been more widely studied, the alterations appearing in acute cardiotoxicity are much less investigated. Thus, our in vivo study aimed to evaluate the process of DOX-induced acute myocardial toxicity by investigating oxidative stress and autophagy markers as mechanisms of myocardial toxicity in correlation with echocardiography and electrocardiography findings. Our results show that both autophagy and oxidative homeostasis were disrupted as soon as 7 days after DOX treatment, alterations that occurred even before the significant increase of NT-proBNP, a clinical marker for cardiac suffering. Moreover, we found a large number of alterations in the electrocardiography and echocardiography of treated rats. These findings suggest that DOX-induced myocardial toxicity started early after treatment initiation, possibly marking the initial phase of the unfolding process of cardiac damage. Further studies are required to completely decipher the mechanisms of DOX-induced cardiotoxicity. Graphical Abstract

Bioactive glasses (BGs), also known as bioglasses, are very attractive and versatile materials that are increasingly being used in dentistry. For this study, two new bioglasses—one with boron (BG1) and another with boron and vanadium (BG2)—were synthesized, characterized, and tested on human dysplastic keratinocytes. The in vitro biological properties were evaluated through pH and zeta potential measurement, weight loss, Ca2+ ions released after immersion in phosphate-buffered saline (PBS), and scanning electron microscopy (SEM) coupled with energy dispersive spectroscopy (EDS) analysis. Furthermore, biocompatibility was evaluated through quantification of lactate dehydrogenase activity, oxidative stress, transcription factors, and DNA lesions. The results indicate that both BGs presented the same behavior in simulated fluids, characterized by high degradation, fast release of calcium and boron in the environment (especially from BG1), and increased pH and zeta potential. Both BGs reacted with the fluid, particularly BG2, with irregular deposits covering the glass surface. In vitro studies demonstrated that normal doses of the BGs were not cytotoxic to DOK, while high doses reduced cell viability. Both BGs induced oxidative stress and cell membrane damage and enhanced NFkB activation, especially BG1. The BGs down-regulated the expression of NFkB and diminished the DNA damage, suggesting the protective effects of the BGs on cell death and efficacy of DNA repair mechanisms.

Background Ovine pulmonary adenocarcinoma (OPA) is caused by Jaagsiekte sheep retrovirus (JSRV) and is considered the most suitable animal model for human lung cancer. The precise mechanisms of OPA oncogenesis are still uncertain. Signal transducer and activator of transcription 3 (STAT3) is activated by interleukin-6 (IL-6) in many cancers, but this aspect is unknown in OPA. We aimed to evaluate the expression of IL-6 and STAT3 in OPA for its potential role in pulmonary carcinogenesis. Results Twenty cases of JSRV-positive OPA and 9 normal lung tissues from sheep were included in the study. Tissue samples were stained with antibodies for IL-6, STAT3, and JSRV-MA. IL-6 and STAT3 were semi-quantified in OPA and control groups using Western Blot (WB). IL‑6 was expressed in stromal, inflammatory, and epithelial cells in all cases of OPA, while STAT3 expression was restricted to epithelial cells. In the OPA group, the percentage of immunolabelled cells for STAT3 accounted for a mean value of 96%. Using the H-SCORE method, 95% of cases were considered positive for STAT3 expression. Control tissuesshowed multifocal andweak immunoexpression for both markers. Using WB analysis, a highly significant amount of both IL-6 (p=0.0078) and STAT3 (pConclusions Our data showed overexpression of IL-6 and STAT3 in OPA in comparison with healthylungs. These results suggest a potential role of IL6-STAT3 in OPA carcinogenesis.

Fungal infections are a growing global health problem. Therefore, our group has synthetized and characterized an improved antimycotic by co-crystallization of ketoconazole and para-amino benzoic acid, named KET-PABA. The aim was to increase bioavailability, biocompatibility, and efficiency of the parent drug-ketoconazole. Based on our previous results showing the cocrystal improved physical properties, such as stability in suspension, solubility, as well as antimycotic efficiency compared to ketoconazole, the current study investigated the local possible side effects induced on the skin of BALBc mice by the application of KET-PABA cocrystal, in view of a further use as a topically applied antimycotic drug. A specific test (mouse ear-swelling test) was used, combined with the histopathological examination and the measurement of pro and anti-inflammatory cytokines and inflammation mediators. KET-PABA application was safe, without signs of skin sensitization shown by the mouse ear sensitization test, or histopathology. KET-PABA strongly inhibited proinflammatory cytokines such as IL1 α, IL1 β, IL6 and TNF α, and other proinflammatory inducers such as NRF2, compared to vehicle. KET-PABA had no effect on the levels of the anti-inflammatory cytokine IL10, or proinflammatory enzyme COX2 and had minimal effects on the activation of the NF-κB pathway. Overall, KET-PABA application induced no sensitization, moreover, it decreased the skin levels of proinflammatory molecules. The lack of skin sensitization effects on BALBc mice skin along with the inhibition of the proinflammatory markers show a good safety profile for topical applications of KET-PABA and show promise for a further clinical use in the treatment of cutaneous mycosis.

Rhinosinusitis is a common disorder related to inflammation of paranasal sinuses and nasal cavity mucosa. Herbal medicines could be an option in the treatment of rhinosinusitis due to their anti-inflammatory and anti-oxidative properties. The study aims to investigate the effect of intranasal Sambucus nigra L. subsp. nigra (SN) extract against inflammation, oxidative stress, and tissue remodeling in nasal and sinus mucosa, but also in serum, lungs, and brain, in Wistar rat model of subacute sinonasal inflammation induced by local administration of lipopolysaccharides (LPS), from Escherichia Coli. The cytokines (TNF-α, IL-1β, IL-6) and oxidative stress (malondialdehyde) in nasal mucosa, blood, lungs, and brain were analyzed. In addition, a histopathological examination was performed, and NF-kB, MMP2, MMP9, TIMP1 expressions were also evaluated in nasal mucosa. Both doses of LPS increased the production of cytokines in all the investigated tissues, especially in the nasal mucosa and blood (p

The study investigated the potential neuroprotective effects of metformin (MET) on alcohol-induced neurotoxicity in adult Wistar rats. The animals were randomized in four groups (n = 10): control, alcohol (ALC), ALC + MET, and MET. ALC (2 g/kg b.w.) and MET (200 mg/kg b.w.) were orally administered for 21 days, once daily. For the ALC + MET group, MET was administered 2 h after ALC treatment. On day 22, the open field test (OFT) and elevated plus maze (EPM) were performed. MET improved global activity and increased the time spent in unprotected open arms, decreased oxidative stress, both in the frontal lobe and in the hippocampus, and increased neuroglobin expression in the frontal cortex. Histopathologically, an increased neurosecretory activity in the frontal cortex in the ALC + MET group was noticed. Thus, our findings suggest that metformin has antioxidant and anxiolytic effects and may partially reverse the neurotoxic effects induced by ethanol.

(1) Background: Photodynamic therapy (PDT) involves the selective killing of tumor cells by the generation of reactive oxygen species using a photosensitizer (PS) activated by irradiation. In melanoma, PDT efficiency is altered by several mechanisms, such as the presence of melanin and melanosomes and pro-survival pathways mediated by transcription factors such as: AP-1 (activator protein), MITF (microphthalmia inducible transcription factor), HIF1α (hypoxia inducible factor), and NF-kB (nuclear factor kappa B). The study aimed to investigate the anti-melanoma effects of PDT mediated by meso-5,10,15,20-tetrakis-(4-hydroxyphenyl)-porphyrin (THPP) as a photosensitizer. (2) Methods: Cocultures of melanoma, two human, WM35 and M1–15, and murine B16-F10, with endothelial cells, were used. Cytotoxicity, oxidative damage, angiogenesis markers, and melanogenesis were assessed using colorimetry, flowcytometry, confocal microscopy, spectrophotometry, ELISA, and Western blotting. (3) Results: The maximal killing efficiency of PDT was reached in WM35, followed by M1–15, and then B16-F10, and it occurred through both apoptosis and necrosis. Although constitutive pigmentation diminished the PDT efficiency, de novo melanogenesis exhibited no protection. PDT increased TNFα, and inhibited NFkB, MITF, HIF1α, and AP1, leading to inflammation and angiogenesis markers’ inhibition. (4) Conclusions: THPP-mediated PDT efficiently induced cell death through apoptosis, necrosis, and the inhibition of pro-survival pathways mediated by NFkB, AP1, HIF1α, and MITF in the melanoma coculture models.

Three nitrogen-doped graphene samples were synthesized by electrochemical exfoliation of graphite rods via pulses of current. Depending on the synthesis conditions, the samples had different content of nitrogen (0.79, 2.56 and 2.33 wt%) and were denoted as NGr-1, NGr-2 and NGr-3, respectively. All samples consist in a mixture of graphene oxide, few- and multi-layer graphene. Several biological effects (cytotoxicity, oxidative stress induction, apoptosis, autophagy and DNA lesions) induced by the N-doped graphenes on human endothelial and colon cancer cells were investigated. The in vitro effects of graphenes were different and depended on the type of cell and the amount of nitrogen in the structure. On human endothelial cells, the graphenes showed antioxidant effects and were less toxic than on colon cancer cells, especially the one with low content of nitrogen (NGr-1). In contrast, on colon cancer cells the graphenes induced cell death by various mechanisms: free radicals generation, apoptosis, autophagy and DNA damage. The most severe DNA lesions and ultrastructural changes were produced by graphene with high content of nitrogen (NGr-2 and NGr-3). Our research demonstrates the cytotoxic effect of graphene with high content of nitrogen on colon cancer cells and antioxidant and protective properties on human endothelial cells.

Background: Oral cancer is highly aggressive due to difficult diagnosis, therapy resistance and increasing frequency; thus finding prevention therapies is very important. Aim: This study evaluates the use of gold and silver nanoparticles (NPs), phyto-synthesized with Cornus mas extract against oral dysplastic lesions. Methods: NPs were characterized by UV–Vis, Fourier-transform infrared spectroscopy, transmission electron microscopy, x-ray diffraction and laser Doppler microelectrophoresis. Biological testing employed two human oral cell lines: gingival fibroblasts and dysplastic keratinocytes and evaluated viability, cell death mechanisms and cellular uptake. Results: NPs induced selective toxic effects against dysplastic cells. p53/BAX/BCL2 activation and PI3K/AKT inhibition led to cell death through necrosis and apoptosis. NPs also induced antioxidant and anti-inflammatory effects. Conclusion: NPs of gold and silver showed promising beneficial effects in the therapy of oral dysplasia.

The study evaluates the effect of adding graphene-Ag nanoparticles (G-AgNp) to a PMMA auto-polymerizing resin, with focus on antibacterial activity, cytotoxicity, monomer release, and mechanical properties. Auto-polymerizing acrylic resin (M) was loaded with 1 wt% G-AgNp (P1) and 2 wt% G-AgNp (P2). Methyl methacrylate monomer release (MMA) was measured after immersion of the samples in chloroform and cell medium respectively. Cell viability was assessed on dysplastic oral keratinocytes (DOK) and dental pulp stem cells. Oxidative stress and inflammatory response following exposure of dysplastic oral keratinocytes to the experimental resins was evaluated. Antibacterial activity against Staphylococcus aureus, Streptococcus mutans and Escherichia coli and also flexural strength of the resins were assessed. Residual monomer: For samples immersed in chloroform, MMA concentration reached high levels, 10.27 μg/g for sample P1; MMA increased at higher G-AgNp loading; 0.63 μg/g MMA was found in medium for P1, and less for sample P2. Cell viability: Both cell lines displayed a viability decrease, but remained above 75%, compared to controls, when exposed to undiluted samples. Inflammation: proinflammatory molecule TNF-α decreased when DOK cultures were exposed to G-AgNp samples. MDA levels indicated increased oxidative stress damage in cells treated with PMMA, confirmed by the antioxidant mechanism activation, while samples containing G-AgNp induced an antioxidant effect. All tested samples showed antibacterial properties against Gram-positive bacteria. Samples containing G-AgNp also exhibited bactericide action on E. coli. Mechanical properties: both samples containing G-AgNp improved flexural strength compared to the sample resin, measured through elastic strength parameters. PMMA resin loaded with G-AgNp presents promising antibacterial activity associated with minimal toxicity to human cells, in vitro, as well as improved flexural properties. These encouraging results obtained in vitro support further in vivo investigation, to thoroughly check whether the PMMA loaded with graphene-silver nanoparticles constitute an improvement over current denture materials.

In recent years, escitalopram (ESC) has been suggested to have different mechanisms of action beyond its well known selective serotonin reuptake inhibition. The aim of this study is to investigate the effects of escitalopram on oxidative stress, apoptosis, brain-derived neurotrophic factor (BDNF), Methyl-CpG-binding protein 2 (MeCP2), and oligodendrocytes number in the brain of chronic unpredictable mild stress-induced depressed rats. The animals were randomised in four groups (8 in each group): control, stress, stress + ESC 5 and stress + ESC 5/10. ESC was administered for 42 days in a fixed dose (5 mg/kg b.w.) or in an up-titration regimen (21 days ESC 5 mg/kg b.w. then 21 days ESC 10 mg/kg b.w.). Sucrose preference test (SPT) and elevated plus maze (EPM) were also performed. ESC improved the percentage of sucrose preference, locomotion and anxiety. ESC5/10 reduced the oxidative damage in the hippocampus and improved the antioxidant defence in the hippocampus and frontal lobe. ESC5/10 lowered caspase 3 activity in the hippocampus. Escitalopram had a modulatory effect on BDNF and the number of oligodendrocytes in the hippocampus and frontal lobe and also improved the MeCP2 expressions. The results confirm the multiple pathways implicated in the pathogenesis of depression and suggest that escitalopram exerts an antidepressant effect via different intricate mechanisms.

Aim To investigate the effects of prenatal exposure to AgNPs obtained by green synthesis with Viburnum opulus L. extract on the testis in male offspring rats. Material and methods Two different doses of AgNPs (0.8 and 1.5 mg/kg b.w.) and vehicle (PBS) were administered to Wistar female rats on days 3–14 of gestation. At 6 weeks after birth, the ultrastructural changes in correlation with the amount of silver as well as the parameters of oxidative stress, inflammation and cell death mechanisms in the testis of male offspring were evaluated. Results AgNPs administered during pregnancy crossed the placental and testicular barriers and induced oxidative stress, DNA damage and autophagy as mechanism of cell toxicity. The markers of inflammation and apoptosis decreased after AgNPs exposure while the NFkB activation increased. TEM examination revealed important ultrastructural changes of Sertoli cells, numerous vacuoles and cytoplasmic changes suggestive of the cellʹs evolution towards necrosis. Conclusion Phytoreduced silver nanoparticles with polyphenols from Viburnum opulus L. fruit extract, administered during the embryological development of the male gonad, have testicular toxic effects in offspring even at 6 weeks after birth.

The novelty of this study consists on the formulation and evaluation of five complex experimental natural photosensitizers (PS): gel with oregano essential oil (O), gel with methylene blue (AM), gel with a mixture of essential oils (Thieves-H), gel with arnica oil and curcuma extract (CU) and gel with frankincense essential oil (T), used as photosensitizing agents (PS) in antimicrobial photodynamic therapy (aPDT) in the control of microbial biofilm in oral cavity. The experimental PS were characterized by gas chromatography-mass spectrometry (GC-MS), Fourier-transform infrared spectroscopy (FTIR), UV-Vis spectroscopy, cytotoxicity assay, antimicrobial effect and scanning electron microscopy (SEM). The IR spectra of the experimental PS with essential oils exhibit absorption bands due to the presence of water and glycerol in high quantities. The studied compounds had a reduced cytotoxic effect on cell cultures. The lowest cytotoxic effect was observed in experimental PS with oregano essential oil and methylene blue PS. Essential oils with proven antibacterial capabilities used in experimental PS confer antibacterial activity to the gels in which they are incorporated, an activity that may be more efficient use of a PDT therapy. Single bacteria were detected mainly by SEM after 12 h, while aggregate bacteria and micro colonies dominated the samples at 48 h.

Aim: To assess ultrastructural changes, alterations in matrix metalloproteinase activity and apoptosis induced by silver nanoparticles (AgNPs) in the rat testicle. Materials & methods: For 45 days, two groups of animals received different doses of AgNPs (0.8 and 1.5 mg/kg b.w.), and a control group was given the buffer used as vehicle for AgNPs. At 7 and 15 days post-treatment, transmission electron microscopy, TUNEL assay, evaluation of NFkB, pNFkB, p53, Bcl-2 and Nrf2 expressions were performed on the removed testes. Results: Transmission electron microscopy revealed severe ultrastructural changes of interstitial tissue and seminiferous epithelium sustained by positive signal for apoptosis. The promatrix metalloproteinase-2 activity and NFkB, Bcl-2 expressions were increased, mainly at 7 days. Conclusion: AgNPs induced severe cell lesions identified even a long time after the exposure.

Cancer nanotechnology is a new field of interdisciplinary research cutting across biology, chemistry, engineering, and medicine, aiming to lead to major advances in cancer treatment. Over the past several years, solid lipid nanoparticles (SLNs) have attracted the interest of researchers due to their ability to overcome the limitations of classic chemotherapeutics. We reviewed the most recent data on the therapeutic use of SLNs in oncology, presenting their main advantages and disadvantages, along with various production methods and different routes of administration. In accordance with these aspects, the long-term physical stability, the controlled release of the loaded drugs, and the efficient targeted delivery of drugs as methods of surpassing the pharmaceutical limitations of anticancer drugs, natural products and gene therapy have been discussed. In addition, we have also emphasized briefly the crosstalk between SLNs and the new trend in oncology, immunotherapy, as future possible antineoplastic treatment, especially in melanoma. This review highlights the potential of SLNs in providing very positive perspectives for future cancer treatment by improving the efficiency of present chemotherapy and reducing its side effects. SLNs allow targeted delivery of anticancer drugs and could improve the efficiency of current chemotherapy in neoplasia.

The aims of our study were to assess the cytotoxic effect of five orthodontic bonding materials, in vitro, on HUVECs to study the induction of apoptosis and inflammatory response generated to assess the shear-bond strength of the 5 tested materials in order to quantify their performance. Standardized samples from each material were obtained and incubated with HUVEC cells for 24 and 72 h immersed in complete medium. Cell viability was determined by means of MTS method. Active caspase 3 and TNFa protein levels were measured through ELISA techniques. The shear-bond strength was tested on 60 extracted premolars which were bonded with the same type of bracket, using the 5 different materials. Statistical analysis Student T-tests, Chi-square and Anova tests were used for results interpretation. Cell viability was decreased with material exposure in a time dependent manner. All materials exerted cytotoxic effects, the experimental materials showed a significantly higher decrease in cell viability at the 72 h reading. Shear Bond strength was superior for the resin commercial bracket-bonding materials. The study shows that orthodontic adhesives� cytotoxicity and physical performance is related to their chemical properties and proves that all orthodontic practitioners should use freely their material of choice on condition they are aware of all its� properties.

Background The antimicrobial activity and effects of a phytocomplex consisting of Tropaeolum flos (T) and Salviae folium (S) extracts on the cytokine levels and transcription factors on dermal fibroblast BJ exposed to bacterial lipopolysaccharides were examined. Methods In order to select the most optimal combination ratio of the two extracts for using in vitro, the physicochemical characterization of vegetal extract mixtures was performed and the antioxidant and antibacterial activities were evaluated on five different formulations of T : S, namely, 1 : 1, 1 : 2, 2 : 1, 3 : 1, and 1 : 3. The best combination of bioactive compounds with regard to antioxidant and antibacterial activities (T : S 1 : 2) was selected for in vitro evaluation of the anti-inflammatory effect. Human dermal fibroblast BJ cells were treated with two doses of the extract mixture and then exposed to bacterial lipopolysaccharides (LPS). The levels of the cytokines involved in inflammatory response, namely, interleukin- (IL-) 6, tumor necrosis factor- (TNF-) α, IL-31, and IL-33, were quantified by ELISA, and the expression of transcription factors, namely, signal transducer and activator of transcription (STAT) 3, nuclear factor kappa B (NFκB), and phosphorylated NFκB (pNFκB), were evaluated by western blot analysis. Results The results have shown that the mixture of T : S 1 : 2 exhibited significant antibacterial effects on Staphylococcus aureus ATCC 25923. LPS exposure increased the cytokine levels in BJ cells and enhanced the NFκB expression. The pretreatment of BF cells exposed to LPS with the two doses of the extract mixture markedly inhibited the increase of IL-33 and TNF-α levels and amplified the NFκB expression and its activation, especially with the high dose. The low doses of the extract reduced NFκB expression but increased its activation. Conclusions These experimental findings suggest that the mixture of T : S 1 : 2 can exert some protection against bacterial infections and inflammation induced by LPS in BJ cells being a good therapeutic option in related conditions associated with inflammation.

Background. The antimicrobial activity and effects of a phytocomplex consisting of Tropaeolum flos (T) and Salviae folium (S) extracts on the cytokine levels and transcription factors on dermal fibroblast BJ exposed to bacterial lipopolysaccharides were examined. Methods. In order to select the most optimal combination ratio of the two extracts for using in vitro, the physicochemical characterization of vegetal extract mixtures was performed and the antioxidant and antibacterial activities were evaluated on five different formulations of T : S, namely, 1 : 1, 1 : 2, 2 : 1, 3 : 1, and 1 : 3. The best combination of bioactive compounds with regard to antioxidant and antibacterial activities (T : S 1 : 2) was selected for in vitro evaluation of the anti-inflammatory effect. Human dermal fibroblast BJ cells were treated with two doses of the extract mixture and then exposed to bacterial lipopolysaccharides (LPS). The levels of the cytokines involved in inflammatory response, namely, interleukin- (IL-) 6, tumor necrosis factor- (TNF-) α, IL-31, and IL-33, were quantified by ELISA, and the expression of transcription factors, namely, signal transducer and activator of transcription (STAT) 3, nuclear factor kappa B (NFκB), and phosphorylated NFκB (pNFκB), were evaluated by western blot analysis. Results. The results have shown that the mixture of T : S 1 : 2 exhibited significant antibacterial effects on Staphylococcus aureus ATCC 25923. LPS exposure increased the cytokine levels in BJ cells and enhanced the NFκB expression. The pretreatment of BF cells exposed to LPS with the two doses of the extract mixture markedly inhibited the increase of IL-33 and TNF-α levels and amplified the NFκB expression and its activation, especially with the high dose. The low doses of the extract reduced NFκB expression but increased its activation. Conclusions. These experimental findings suggest that the mixture of T : S 1 : 2 can exert some protection against bacterial infections and inflammation induced by LPS in BJ cells being a good therapeutic option in related conditions associated with inflammation.

Background Photodynamic therapy (PDT) is a treatment of cancer due to its ability to induce cell death, oxidative stress and acute inflammatory reaction in targeted sites. To optimize the effect of PDT the addition of some compounds with supplementary cytotoxic effect on tumor cells was tried. Methods The study was performed on 35 Wistar male albino rats with Walker 256 carcinosarcoma. The animals were randomly assigned in seven groups (n = 5) and treated as follows: group 1 - control; group 2 – Cornus mas (CM) extract 15 mg/kg b.w., administered for 7 days; group 3 – CM extract administered for 7 days followed by irradiation (CM + IR); group 4 – one dose of tetra-p–sulfonato-phenyl-porphyrin (TSPP) 10 mg/kg b.w.; group 5 – TSPP + IR; group 6 – CM extract administered daily for 7 days before TSPP and IR (CM + TSPP + IR); group 7 – TSPP + IR followed by CM administered for 7 days (TSPP + IR + CM). Results The results showed that MDA and GSSG levels increased after PDT in parallel with the increasing of COX-2 expression and DNA damage. Apoptotic and necrotic index enhanced in TSPP + IR, effect improved by CM association before PDT. CM + TSPP + IR regimen also induced more intense inflammatory reactions, increased COX-2 expression, determined DNA damage, apoptosis and necrosis, compared to the TSPP + IR + CM group. Both combined therapeutic regimens reduced MDA levels in tumor tissue, especially CM + TSPP + IR and increased the antioxidant defense and iNOS expression. Conclusions Our results demonstrated that CM associated before PDT had beneficial effects in PDT and may represent a promising option in PDT strategies.

The study aims to evaluate the impact of silver nanoparticles, phytosynthesized with polyphenols from Sambucus nigra L. (SN) fruit extract (AgSN), on dysplastic oral keratinocytes (DOK) and human gingival fibroblasts (HGF) in terms of cell viability and apoptosis. The morphology and ultrastructure of treated cells as well as the mechanisms involved in cell death induction were investigated in DOK cultures. The structure of AgSN was studied by using the appropriate analysis tools such as UV–Vis, transmission electron microscopy, Raman spectroscopy, dynamic light scattering (DLS) and zeta potential assessment. DOK and HGF were treated either with silver nanoparticles capped with Sambucus nigra L. extract or with SN extract. Untreated cells were used as controls. Viability was determined by MTS assay. Transmission electronic microscopy (TEM) was used to evaluate the intracellular localization of the nanoparticles at 4 and 24 h. Annexin V-FITC/propidium iodide staining and the expressions of p53, BAX, BCL2, NFkB, phosphorylated NFkB (pNFkB), pan AKT, pan phosphoAKT, LC3B and ɣH2AX were evaluated to quantify the cell death. ELISA measurements of TNF-α and TRAIL was used for the study of the inflammatory response. Oxidative stress damage induced by nanoparticles was assessed by the malondialdehyde (MDA) level. Silver nanoparticles stimulated HGF proliferation and significantly diminished DOK viability at doses higher than 20 μg/ml. TEM analysis demonstrated the internalization of silver nanoparticles and showed ultrastructural changes of cells such as the appearance of vacuoles, autophagosomes, endosomes. AgSN inhibited the pro-survival molecules and regulators of apoptosis, diminished oxidative stress and inflammation and induced cell death through various mechanisms: necrosis, autophagy and DNA lesions. SN extract had antioxidant and anti-inflammatory effect and increased the DNA lesions and autophagy in DOK cells. Silver nanoparticles protected the normal cells and induced cell death in dysplastic cells by different mechanisms thus offering beneficial effects in the treatment of oral dysplasia.

Melanoma, a cancer derived from melanocytes is very difficult to treat, especially in advanced cases. There are several encouraging studies of the efficacy of photodynamic therapy (PDT) in melanoma. However, PDT has to overcome the main defense mechanisms like: defects in the apoptotic pathways, pigmentation, sequestration of the photosensitisers (PS) inside melanosomes and increased oxidative stress defense. Two meso-substituted porphyrins, meso-5,10,15,20-tetrakis (4-hydroxyphenyl) porphyrin (THOPP) and meso-5-(4-hydroxyphenyl)-10, 15, 20-tris (4-methoxyphenyl) porphyrin (THOMPP) were used as PS to investigate several mechanisms underlining the PDT anti-melanoma effects, on a lightly pigmented melanoma cell line (WM35), in vitro. γH2AX foci formation (a measure of DNA double strand brakes) was used for the assessment of DNA damage by means of immune-fluorescence and western blot. Cytoskeleton alterations were detected by phalloidin staining. Tyrosinase activity and melanin pigment were quantified by spectrophotometry, tyrosinase protein by western blot, total peroxidase activity by resorurfin reaction (Amplex Red). PDT induced high levels of DNA damage, cytoskeleton alterations and enhanced pigmentogenesis. THOPP mediated PDT was the most efficient. The melanogenesis stimulated by PDT was directly correlated to the PDT induced cellular damage and provided no protection against therapy. Thus, PDT induced melanogenesis combined with severe DNA damage was able to overcome the mechanisms of resistance and increased the efficiency of PDT in WM35 melanoma cells. These results are encouraging for a possible use of PDT, as an adjuvant therapy in lightly pigmented melanomas.

The present study presents a green, cost efficient and easy synthesis method of silver nanoparticles (AgNPs) using an aqueous extract of Cornus sanguinea L. fruits (CS). The phytosynthesized silver nanoparticles were characterized using various analytical techniques such as UV–Vis absorption spectroscopy, which confirmed the formation of AgNPs and FTIR spectroscopy, in order to certify the role of the biomolecules present in the fruit extract as reducing and capping agents of the AgNPs. The UV–Vis absorption spectrum showed a broad band at 407 nm characteristic for colloidal silver. Transmission electron microscopy was conducted to investigate the shape and size of the silver nanoparticles, revealing a spherical shape with an average particle size of 18 nm. The antioxidant and anti-inflammatory activities of the fruit extract and green synthesized silver nanoparticles were assessed in vivo on experimental inflammation. The obtained results showed that CS and AgNPs reduced oxidative stress in parallel with increasing of antioxidant defense and diminished the COX-2 expressions. CS extract had a dual effect on NFkB activation depending on the time of testing while AgNPs increased NFkB phosphorylation at 48 h. These results suggested that both AgNPs and CS extract exhibited antioxidant and anti-inflammatory activities but with a different dynamics of action.

The purpose of our study is to investigate the comparative effects of materials based on silver and gold nanoparticles functionalized with polyphenols from Cornus Mas extract (AgNPs-CM and AuNPs-CM) in vivo on experimental inflammation. The nanoparticles were obtained at room temperature under UV irradiation and were characterized by different methods: ultraviolet-visible spectroscopy, transmission electron microscopy, X ray diffraction, Fourier transform infrared spectroscopy and dynamic light scattering. The modulatory effects of AgNPs-CM and AuNPs-CM on inflammation were quantified by oxidative stress parameters, pro and anti-inflammatory cytokines levels and apoptosis assessment at 2 h, 24 and 48 h after induction of inflammation with carrageenan in the paw tissue of Wistar rats. Our results showed that silver and gold nanoparticles only partial and for a short period have mobilized the antioxidant defense mechanisms. In addition, they diminished inflammation and apoptosis in the early stage while later, at 48 h, exerted an immunomodulatory effect, activated ERK ½ and induced apoptosis. The photoreduced silver and gold nanoparticles, functionalized with natural compounds, modulated the inflammation in a similar manner in the soft tissue injected with carrageenan. In order to decipher the mechanisms involved in interactions of metallic nanoparticles with biological systems and for a complete assessment of the risks and benefits of these products in clinical practice long term studies are necessary.

Photodynamic therapy (PDT) could be an adjuvant therapy in melanoma, an aggressive cancer that arises from melanocytes. Several reports showed encouraging results of the efficacy of PDT in melanoma on experimental models and in clinical trials. Therefore, we studied the efficacy of two derivatives of tetraphenylporphyrin (TPP): meso-5,10,15,20-tetrakis (4-hydroxyphenyl) porphyrin (THOPP) and meso-5-(4-hydroxyphenyl)-10,15,20-tris (4-methoxyphenyl) porphyrin (THOMPP) as photosensitizers for PDT, compared to FDA approved delta aminolevulinic acid (ALA) against a lightly pigmented, melanoma cell line, WM35, in vitro. Both porphyrins were more efficient as photosensitizers, compared to ALA, without dark toxicity. The efficiency depended on the intracellular localization and the molecule structure. THOPP, the most efficient porphyrin localized mainly in mitochondria, while THOMPP accumulated in lysosomes; both showed melanosomal localization. The symmetric THOPP molecule was able to generate increased oxidative stress damage and apoptosis. THOPP also induced a low effect on the defense mechanisms like antioxidant enzyme SOD (superoxide dismutase), NF-kB (nuclear transcription factor kB) activation and MITF (microphthalmia transcription factor). The lower efficiency of the asymmetric molecule, THOMPP was probably due to a diminished photoactivation, which led to a lower ROS induced damage, combined with higher activation of the defense mechanisms.

Melanoma, a cancer that arises from melanocytes, is one of the most unresponsive cancers to known therapies and has a tendency to produce early metastases. Several studies showed encouraging results of the efficacy of photodynamic therapy (PDT) in melanoma, in different experimental settings in vitro and in vivo, as well as several clinical reports. Aims. Our study focuses on testing the antimelanoma efficacy of several new, synthetic photosensitisers (PS), from two different chemical classes, respectively four porphyrins and six phthalocyanines . Methods. These PS were tested in terms of cell toxicity and phototoxicity against a radial growth phase melanoma cell line (WM35), in vitro. Cells were exposed to different concentrations of the PS for 24h, washed, then irradiatied with red light (630 nm) 75 mJ/cm 2 for the porphyrins and 1 J/cm 2 for the phthalocyanins. Viability was measured using the MTS method. Results. Two of the synthetic porphyrins, TTP and THNP, were active photosensitizers against WM35 melanoma in vitro. Phthalocyanines were effective in producing a dose dependent PDT-induced decrease in viability in a dose-dependent manner. The most efficient was Indium (III) Phthalocyanine chloride, a metal substituted phthalocyanine. Conclusions. The most efficient photosensitizers for PDT in melanoma cells were the phthalocyanines in terms of tumor cell photokilling and decreased dark toxicity.

Background. The antimicrobial activity of theEquisetum arvenseL. extract and the mechanisms involved in thein vitroeffects on endothelial vascular cells exposed to hyperosmotic stress were evaluated.Methods. Antimicrobial activity was evaluated by disk diffusion method and minimum inhibitory concentration (MIC) determination, and oxidative stress, inflammation, and apoptosis, in pretreatment withEquisetum arvenseL., caffeic acid, and cathechin, were quantified.Results. The results have shown thatEquisetum arvenseL. exhibited antibacterial effects only on pathogenic gram-positive cocci.The modulatory activity ofEquisetum arvenseL. on endothelial cells exposed to hypertonic medium was different and depended on the concentration used. Low concentrations of tested compounds exerted antioxidant effect and diminished the activity of caspase-8 and also increased IκB expression while in high doses,Equisetum arvenseL. was prooxidant, induced apoptosis, and decreased IL-6 secretion.Conclusions. These experimental findings suggest thatEquisetum arvenseL. has antibacterial effects on gram-positive cocci and, administered in low dose, may be a new therapeutic approach for diseases associated with hypertonic conditions or oxidative stress and apoptosis.

Finding appropriate experimental designs and analysis methods in order to gain insight into the mechanisms of efficiency and toxicity of nanomaterials is a major focus in today’s research in nanomedicine. In this paper, we demonstrate the ability of scanning confocal Raman spectroscopy to emphasize the molecular changes in terms of inflammation resolution after administration of a single dose of metal nanoparticles functionalized with natural extracts, in experimental inflammation. Five experimental groups of Wistar rats were used, treated with one dose of gold nanoparticles (AuNPs–CM), one dose of silver nanoparticles (AgNPs–CM), Cornus mas (CM) extract, and vehicle, before intraplantar injection of 100 μL 1% carrageenan, and one group of untreated animals. The paw tissues were harvested and used for transmission electron microscopy (TEM) and evaluation of the metal content 4 and 24 hours after the induction of inflammation and, after 24 hours, Raman spectroscopy, histopathology and prostaglandin (PG) E2 level assessment were performed. TEM revealed varying degrees of alterations in dermo-epidermal junctions and capillaries, especially in tissues treated with AgNPs–CM and vehicle, in parallel with the increase in PGE2 levels. Besides ultrastructural changes highlighted by TEM, meaningful information about the molecular changes is provided by multivariate Raman spectral images. Indeed, thorough Raman spectral analysis shows that AuNPs–CM and CM restored the normal composition of unsaturated fatty acids while the specimens treated with AgNPs–CM were dominated by the protein component. Our results suggest that the Raman spectral analysis has real potential to be used in tandem with standard methods for monitoring the subtle molecular effects induced by the administration of nanoparticles.

Aim: To investigate the comparative effects of prenatal exposure to silver nanoparticles (AgNPs) functionalized with citrate and polyphenols on spatial cognition and also on nitro-oxidative stress and apoptosis in the hippocampus and cerebellum of offsprings. Materials & methods: Pregnant Wistar rats were orally administered substances from day 10 of gestation until day 21. Six weeks postpartum, six male offsprings from each group were used for behavioral evaluation and determination of oxidative stress and apoptosis. Results: Both groups exhibited hyperactivity and anxiety especially after AgNPs-Sambucus nigra L. administration. AgNPs-S. nigra L. group showed increase in induced nitric oxide synthase activity and decrease in superoxide dismutase activity and apoptosis in the hippocampus, while AgNPs-citrate coated administration exerted a moderate toxicity and induced apoptosis. Conclusion: AgNPs functionalized with natural extracts had a lower toxicity than citrate-coated silver particles.

Introduction Exposure of the endothelial cells to hypoxia, the decrease in oxygen supply can trigger an endothelial response. This response is involved in inflammatory diseases, tumorigenesis, and also with the micro vascular damage associated with aging. The aim of our study was to determine the hypoxia/re-oxygenation induced response in vitro, using human umbilical vein endothelial cells (HUVEC) cultures, at different time points with focus on cell viability, apoptosis oxidative stress and angiogenesis stimulation. Materials and methods Cells were exposed to 10%, 5% or 0% O2 for 6 h, 12 h, and 24 h. Viability was measured through colorimetry, apoptosis – annexin V-FITC staining, DNA lesions (γH2AX), endothelial activation (sICAM1), angiogenesis (HIF1α), oxidative stress (malondialdehyde, superoxidismutase and NFκB activation) were determined by ELISA, Western Blot and spectrophotometry. Results and discussion Hypoxia decreased viability, increased apoptosis, oxidative stress, endothelial activation and angiogenesis, depending on O2 concentration and time exposure. Short exposures to 5% and 10% O2, efficiently activated anti-apoptotic mechanisms through NFκB activation, HIF1α and γH2AX related DNA damage repair pathways. However, severe hypoxia and longer exposures to mild hypoxia induced high oxidative stress related damage and eventually led to apoptosis, through strong increases of HIF1α and accumulating DNA lesions.