Search

The present study aimed to assess the feasibility of developing low-cost multipurpose iron oxide/TiO

There is a fine balance between inflammation and tumorigenesis. While environmentally induced inflammatory condition can precede a malignant transformation, in other cases an oncogenic change of unknown origin can induce an inflammatory microenvironment that promotes the development of tumors. Regardless of its origin, maintaining the inflammation milieu has many tumor-promoting effects. As a result, inflammation can aid the proliferation and survival of malignant cells, can promote angiogenesis and metastasis, can down-regulate innate/adaptive immune responses, and can alter responses to hormones and chemotherapeutic agents. There is an abundance of studies unveiling molecular pathways of cancer-related inflammation; this wealth of information brings new insights into biomarkers domain in the diagnosis and treatment improvement pursue. In cutaneous tissue there is an established link between tissue damage, inflammation, and cancer development. Inflammation is a self-limiting process in normal healthy physiological conditions, while tumorigenesis is a complex mechanism of constitutive pathway activation. Once more, in cutaneous melanoma, there is an unmet need for inflammatory biomarkers that could improve prognostication. Targeting inflammation and coping with the phenotypic plasticity of melanoma cells represent rational strategies to specifically interfere with metastatic progression. We have shown that there is a prototype of intratumor inflammatory infiltrate depicting a good prognosis, infiltrate that is composed of numerous T cells CD3+, Langerhans cells, few/absent B cells CD20+ and few/absent plasma cells. Circulating immune cells characterized by phenotype particularities are delicately linked to the stage melanoma is diagnosed in. Hence circulatory immune sub-populations, with activated or suppressor phenotype would give the physician a more detailed immune status of the patient. A panel of tissue/circulatory immune markers can complete the immune status, can add value to the overall prognostic of the patient and, as a result direct/redirect the therapy choice. The future lies within establishing low-cost, affordable/available, easily reproducible assays that will complete the pre-clinical parameters of the patient.

Phthalocyanines (Pc) and their metallated derivatives are strongly considered for photodynamic therapy (PDT) possessing unique properties as possible new photosensitizers (PS). We have used toxicological assessments, real‐time monitoring of cellular impedance, and imagistic measurements for assessing the in vitro dark toxicity and PDT efficacy of Ga(III)‐Pc in SHSy5Y neuroblastoma cells. We have established the non‐toxic concentration range of Ga(III)‐Pc, a compound which shows a high intracellular accumulation, with perinuclear distribution in confocal microscopy. By choosing Ga(III)Pc non‐toxic dose, we performed in vitro experimental PDT hampering cellular proliferation. Our proposed Ga(III)‐Pc could complete a future PS panel for neuroblastoma alternate therapy.

Neoplastic disease has multifactorial etiology and insidious evolution which make it unlikely to be detected in early stages and very difficult to treat at later times. The effectiveness of standard therapeutic approaches is limited by severe adverse effects, metastasis, and tumor capacity to develop multidrug resistance.

Recently, a large spectrum of biomaterials emerged, with emphasis on various pure, blended, or doped calcium phosphates (CaPs). Although basic cytocompatibility testing protocols are referred by International Organization for Standardization (ISO) 10993 (parts 1–22), rigorous in vitro testing using cutting-edge technologies should be carried out in order to fully understand the behavior of various biomaterials (whether in bulk or low-dimensional object form) and to better gauge their outcome when implanted. In this review, current molecular techniques are assessed for the in-depth characterization of angiogenic potential, osteogenic capability, and the modulation of oxidative stress and inflammation properties of CaPs and their cation- and/or anion-substituted derivatives. Using such techniques, mechanisms of action of these compounds can be deciphered, highlighting the signaling pathway activation, cross-talk, and modulation by microRNA expression, which in turn can safely pave the road toward a better filtering of the truly functional, application-ready innovative therapeutic bioceramic-based solutions.

We report results regarding the in vitro toxicology of γ-Bi2O3 represented by its isomorphous phase Bi12TiO20 (γ-BTO). The γ-BTO microparticles were synthesized by two methods: coprecipitation from a bismuth nitrate–tetrabutyl titanate solution and solid state reaction of Bi2O3 and TiO2 oxides. The structural and morphological characteristics of the obtained materials were determined using X-ray diffraction (XRD), selected area electron diffraction (SAED), transmission (TEM) and scanning (SEM) electron microscopy. The elemental composition was investigated using energy dispersive spectrometry (EDS). The cytotoxicity and oxidative/nitrosative stress (intracellular reactive oxygen species (ROS) and nitric oxide (NO) release) induced by the studied microparticles in HepG2, SH-SY5Y and 3T3-L1 cell cultures were determined using the MTT, DCF-DA (2′,7′-dichlorfluorescein-diacetate) and Griess methods respectively. Depending on the cell type and γ-BTO concentration, results showed only weak cytotoxic effects after 24 h of γ-BTO exposure and cell proliferation effects for longer treatment times. Only reduced NO release increases (corresponding to high γ-BTO concentrations) were detected in case of SH-SY5Y and 3T3-L1 cells. The intracellular ROS production (higher for HepG2 cells) appeared inversely proportional to the γ-BTO concentration. The obtained results indicated a promising in vitro biocompatibility of γ-BTO and encourage further studies regarding its potential for biomedical applications.

We report results of noncellular tests, revealing the occurrence of photocatalytic interactions between titanium dioxide (TiO2, titania) nanoparticles and the MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide] cytotoxicity indicator. These interactions induce the reduction of MTT and formation of purple formazan under biologically relevant conditions. Classical MTT assays have been performed to evaluate the production of formazan in DMEM-F12 and RPMI-1640 cell culture media (containing 10% fetal bovine serum-FBS) treated with Degussa-P25 TiO2 nanoparticles, in the absence of cells. The colorimetric determinations revealed the noncellular MTT to formazan transformation induced by TiO2 nanoparticles, under conditions commonly used for in vitro cytotoxicity testing of nanomaterials. The formazan precipitation was found to be proportional to the TiO2 concentration, being enhanced under laboratory daylight exposure. The photocatalytic nature of the studied effect was assessed under UV irradiation at 365nm. The biological significance of the reported reaction was established with respect to cellular reference experiments performed on V79-4, HeLa and B16 cell lines. The results show false viability increases with up to 14% (for TiO2 concentrations generally higher than 50μg/ml), induced by the TiO2-MTT reaction. This type of artifacts may lead to underestimated toxicity or false proliferation results.

The aim of this study was to obtain a novel hydroxyapatite-based material with high biocompatibility. The structural properties of the samples were well characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and X-ray Photoelectron Spectroscopy (XPS). The X-ray diffraction studies revealed the characteristic peaks of hydroxyapatite in each sample. Other phases or impurities were not observed. The scanning electron microscopy observations suggest that the doping components have no influence on the surface morphology of the samples, which reveals a homogeneous aspect of the synthesized particles for all samples. The presence of calcium (Ca), phosphor (P), oxygen (O) and silver (Ag) in the Ag:HAp is confirmed by energy dispersive X-ray (EDAX) and X-ray Photoelectron Spectroscopy analyses. Nanocrystalline silver doped HAp stimulated viability and potentiated the activation of murine macrophages.

In the context of an emerging market of food supplements, the proven quality of the antioxidant products should be the main criteria for using them. The production process has to be carefully controlled and complementary tests are needed to demonstrate the correspondence between real and declared properties of final product. Using well characterized compounds with proven antioxidant activity in biological systems as reference brings a plus of rigorously to the testing protocol. The aim of this study was to determine the acceptance range for the antioxidant (peroxyl radicals scavenger) capacity of "Natural SOD" by using for comparison ascorbic acid (vitamin C). The established acceptance range complete our previous results concerning the antioxidant capacity of Natural SOD using validated ORAC method and creates premises for supplementary checking of the batches in the current production and improving the product quality.

AIM: The aim of the study described in the present paper was to assess several in vitro effects of TiO 2 nanoparticles with different colloidal and photocatalytic properties on RAW 264.7 macrophages. METHODS: The cells were exposed to Degussa P25 titania and two other types of nanoparticles synthesized by a hydrothermal procedure in our laboratory: undoped and Fe 3 +-doped TiO 2 . Compared to Degussa P25, the hydrothermal nanomaterials were significantly less active in inducing cytotoxicity, production of intracellular reactive oxygen species (ROS) and release of pro-inflammatory cytokine interleukin-6 (IL-6). The induced effects were analysed with respect to nanoparticle size, surface charge, hydrophilicity, semiconductor bandgap energy and photocatalytic generation of ROS under non-cellular conditions. RESULTS: The overall results indicated that TiO 2 nanoparticles with higher surface charge, hydrophilic surfaces and enhanced photocatalytic properties may preferentially induce macrophage cell damage and inflammation compared to other TiO 2 nanomaterials. CONCLUSION: The present findings are relevant for studies regarding the evaluation of risks raised by self-cleaning technologies involving nanosized hydrophilic TiO 2 photocatalysts as well as development of synthesis methods optimized for producing biocompatible TiO 2 nanomaterials.

This paper presents a new device generating high-voltage pulsed cold atmospheric plasma jets. With these plasmas, the quantity of atomic oxygen (and, accordingly, the chemical activity) is a lot higher than that in previous researches. The main characteristic of the new device is the usage of three tubular needle-type electrodes connected in parallel. By applying high-voltage pulses (with 20-30-kV amplitude, duration of hundreds of nanoseconds, and hundreds of pulses per second), three independent discharges are formed in the discharge room. The plasma-forming gas is He, with a low quantity of oxygen introduced through the high-voltage electrodes. Each discharge has an optimal percentage of 0.5 % vol. O2 in He, which maximizes the quantity of atomic oxygen in the plasma. The plasmas of the three discharges unite to a single jet though, which contains a quantity of atomic oxygen that is a lot higher than that of the plasma of each discharge. The emission spectra of the plasma jets show the maximization of the intensity of the O I 777 nm line when a concentration of 1.5% vol. O2 in He (three times higher than so far) is introduced in the plasma-forming gas.

High-voltage pulsed cold atmospheric plasma jets were used to treat tumoral (B16 and COLO320) and normal (macrophage) cells. Cold atmospheric plasma jet generator and plasma jet physical characteristics are presented. The treatment of the tumoral cells intended to obtain their apoptosis. For tumoral cells, percents of up to 70% apoptosis have been obtained, using helium plasma, activated with up to 2% oxygen. In the same experimental conditions, the plasma treatment did not induce apoptosis in normal (macrophage) cells. In addition, the macrophage cells were not activated by the plasma jets.

Influence of the novel arthritis drug-substance MCS-18 on the antibody (Ab) production against tetanus toxoid (TT) and diphtheria toxoid (DT) antigens was tested in vivo. Possible involvement of MCS-18 in Toll-like receptor (TLR) signalling pathway was further considered.Immunization of male CD1 mice was done with subcutaneous injection of TT emulsified in Freund's Complete (FCA) or Incomplete Adjuvant (FIA) and mixed diversly with MCS-18 and different test substances. To investigate the influence of TLR activation Pam3Cys and lipopolysaccharide (LPS) emulsified in FIA were tested in combinations with MCS-18. Antibody production was analysed in vivo by tetanus- or diphtheria-toxin neutralization test.Immunogenicity of TT was significantly enhanced if administered together with FCA or TLR agonists Pam3Cys or LPS emulsified in FIA. It was shown that MCS-18 attenuated strongly the production of anti-TT Ab if administered together with the Ab elicitor FCA or TLR agonists in various combinations. MCS-18 was also active via oral administration.These findings suggest that MCS-18 could be a potent, non-toxic antagonist or a down-regulator of TLR signalling pathway. Investigations on further models are needed to establish ifMCS-18 may influence particularly the production of RA-specific auto-antibodies, too.

High-mobility group box protein 1 (HMGB1) is an intracellular protein that may be released actively from monocytes and macrophages or passively from necrotic or damaged cells. Its inhibition in animal experiments, even in the late phase of septic shock, significantly enhanced the survival rate of rodents. The aim of our study was to investigate the effect of a vegetal fraction isolated and highly purified from Helleborus purpurascens regarding the modulation of HMGB1 release either from tumor cells or human blood mononuclear cells. Our results showed that the vegetal fraction was able to down-regulate the release of HMGB1 from activated human blood mononuclear cells (PBMCs) and tumor cells. By combining the purified fraction with Cyclophosphamide the release of HMGB1 from tumor cells was strongly decreased. This synergism was not noticed when the ve getal product was associated with Doxorubicin. We also studied the effect of the purified fraction in mice with septic shock induced by cecal ligation and puncture (CLP) method. The tested vegetal product increased significantly the survival rate of animals compared to the mice not treated with it. Our data suggest that the purified vegetal fraction may modulate inflammation by down-regulating the HMGB1, which can also explain its efficacy in septic shock in mice.

The cells of immune system such as monocytes and macrophages are in first line defence against dangerous signals. In the present paper the recognition of Dectin 1 receptors and the modulation of Interleukin-10 (IL-10) and Tumor Necrosis Factor-alpha (TNF-alpha) cytokine production by Curdlan and Curdlan derivatives in peripheral blood mononuclear cells (PBMCs) were studied. The effect of Curdlan or Curdlan derivatives on the expression of Dectin 1 receptors in PBMCs was revealed by flow-cytometry and the levels of IL-10 and TNFalpha were measured by ELISA kit in supernatants of PBMCs cultured in presence or absence of Curdlan, Curdlan derivatives and LPS. Our results suggested that Curdlan and Curdlan derivatives were able to increase the expression of Dectin-1 receptors on monocyte cells. The combined treatment of Curdlan/Curdlan derivatives and Pam3Cys produced an increase of CD14+ cells possessing Dectin-1 receptors. We demonstrated that Curdlan (at 20 microg unique dose) up-regulated TNF-alpha production and down-regulated IL-10 production in PBMCs. Conversely, Palm CM/SP-Curdlan (20 microg unique dose) was able to down-regulate TNF-alpha production and to up-regulate IL-10 production in PBMCs. For instance, Palm CM/SP-Curdlan determined a 5 times decrease of TNF-alpha production than Curdlan. Regarding the effect of Palm CM/SP-Curdlan on IL-10 production in PBMCs, we noticed that the level of IL-10 was about 4 times greater than Curdlan activity. We observed that a combined treatment of Curdlan/Curdlan derivatives and LPS induced about 5 times decrease in TNF-alpha production in PBMCs. IL-10 production induced by Palm CM/SP-Curdlan and LPS was about 6 times greater than the combined effect of Curdlan and LPS. The treatment of PBMCs with SP-Curdlan alone affected neither TNF-alpha production nor IL-10 production. Our results are in accordance with other studies demonstrating that Dectin-1 and TLR2/TLR6 signaling combine to enhance the responses triggered by each receptor and the signaling pathway induced by Dectin-1 could mediate the production of pro-inflammatory cytokines.

The effects of cold plasmas are due to charged particles, reactive oxygen species (ROS), reactive nitrogen species (RNS), UV photons, and intense electric field. In order to obtain a more efficient action on mammalian cells (useful for cancer therapy), we used in our studies chemically activated cold plasma (He and O2 gas mixture). V79-4 cells were exposed to plasma jet for different time periods (30, 60, 90, 120 and 150s), using different combinations of helium and oxygen inputs (He:2.5l/min + 02:12.5ml/min; He:2.51/min + O2:25ml/min; He:2.51/min + O2:37.5 ml/min). Using MTT test we demonstrated that plasma jet induced cell viability decrease in all cases. The effect of chemically activated cold plasma--apoptosis or necrosis--depends on gas mixture and treatment period. Taking into account that ROS density in cell microenvironment is related to O2 percent in the gas mixture and treatment period, we can presume that cell death is due to ROS produced in plasma jet.

The chemotherapy success to kill cancer cells depends on its ability to stop cell division. The faster the cells are dividing, the more likely it is that chemotherapy will kill the cells, causing the tumor to shrink. Taking into account the severe side effects of chemotherapy, drugs producers also focus on natural products obtained either from medicinal plants, or from microorganisms. The complex polysaccharides named beta-glucans are active compounds with immune activity. beta-glucan polymers belong to a class of drugs with effects on the immune system, such as: anti-tumoral, anti-infectious, protection against fungi, bacteria and viruses infections. The correct selection of beta-glucans is essential to identify compounds with favorable clinical effects. The aim of this study was to investigate the capacity of six Curdlan (beta-glucan) derivatives to up-regulate the Doxorubicin, Actinomycin D and Cyclophophamide cytostatic drug activity on tumor cells (murine B16 melanoma and human HEp-2 laryngeal carcinoma cell lines). Our results demonstrated that Palm SP derivative, as well as SP and Palm CM/SP derivatives were able to potentiate Doxorubicin action or Actinomycin D effect on B16 tumor cells. SP derivative significantly enhanced cytostatic activity of Cyclophosphamide on B16 cells. All the investigated Curdlan derivatives (SP, Palm CM/SP, CM/SP, Palm CM, Palm SP and CM) were able to inhibit HEp-2 tumor cell growth, by up-regulating Doxorubicin and Actinomycin D cytostatic activity.

Cold atmospheric plasma treatment acts at the cellular level to remove diseased tissue without inflammation and damage, to suppress infections and to modulate the viability (apoptosis/necrosis) of tumoral cells. It is also known that, a major cause of anti-tumor chemotherapy failure is the development of multidrug resistance (MDR) of tumors. This study reveals the effect of high voltage pulsed, repetitive cold atmospheric plasma jets which are chemically activated with oxygen, on B16 tumoral cells (murine melanoma cell line) and COLO320DM multidrug resistant cells (human colon cancer cell line). The tests have been performed on human colon cancer cell line COLO320DM and murine melanoma cell line B16-F10. These cell lines have been treated with cold helium or helium-oxygen generated plasma jets and the consequent apoptosis has been analyzed by means of flow cytometric method. A treatment time-dependent apoptosis has been observed only in the case of 816-F10 cells interacting with helium-oxygen plasma and no apoptosis has been identified when the cells were treated only with helium plasma jets. These results indicate the need of oxygen for the chemical activation of plasma. The COLO320DM cells (that over-express the MDR efflux pumps) have been exposed to helium-oxygen plasmas only, or in a combination with vegetal extract MCS D161 as MDR efflux pumps inhibitor. For the secondly mentioned case the results have showed an increased apoptosis rate compared to the plasma treatment alone. The obtained data represent a starting point for the study of a possible combined treatment (atmospheric pressure cold plasmas and a MDR efflux pumps inhibitor applied with chemotherapy).

Reactive oxygen species (ROS) are well known to be cytotoxic and have been implicated in the etiology of a wide array of human diseases including diabetes, neurodegenerative diseases, cancer and also influence central cellular processes such as proliferation, apoptosis, senescence etc. If in these pathological or degenerative conditions characterized by free radicals excess, reactive species are not eliminated, they can maintain destructive processes, already initiated at different cellular levels. Understanding the role of ROS as key mediators in signaling cascades may provide various opportunities for pharmacological intervention. Toll-like receptors and C-type lectin receptor class V--Dectin-1, as members of Pattern Recognition Receptors play an essential role in innate immune response against bacteria and fungi respectively, contributing to pathogens recognition, phagocytosis, ROS production and induction of pro-inflammatory cytokines secretion. Using a high performance chemiluminometric method, we studied the action of six Curdlan derivatives on the ROS production and release by activated human polymorphonuclear cells (PMNs) isolated from the peripheral blood of healthy donors. Our results demonstrated that Curdlan derivatives containing sulfopropyl groups did not activate human PMNs to release ROS. These compounds blocked Dectin-1 and were able to inhibit co-operation between Dectin-1 and TLR-2. Curdlan derivatives containing palmithoyl, carboxi-methyl and sulfopropyl groups increased ROS release by human PMNs activated at TLR-2 level. Taking into account the fact that Dectin-1 can actively collaborate with TLR-2 to modulate the subsequent adaptive immune response, we can presume that Curdlan derivatives containing sulfopropyl group or palmithoyl/carboxi-methyl/sulfopropyl groups, as possible Dectin-1 antagonists/agonists, could influence TLR-2 signaling.

Toll-like receptors (TLRs) and Dectin-1, as members of Pattern Recognition Receptors play an essential role in innate immune response against bacteria and fungi respectively, contributing to pathogens recognition, phagocytosis, etc. Dectin-1 and TLR-2/TLR-6 can interact for intracellular signal transduction. Dectin-1 is expressed at low levels on macrophages and at high levels on dendritic cells. Dectin-1 and TLRs are synergistic in mediating cytokines production, such as IL-12 and tumor necrosis factor alpha (TNF alpha). In the present paper we studied the expression of Dectin-1 (beta-Glucan Receptor C-type lectin receptor class V) and TLR-2 on human normal monocytes cells and also the role of different Curdlan derivatives and highly purified natural extracts, especially their capacity to recognize these receptors and their Dectin-1 agonist/antagonist properties. Our results demonstrated that Curdlan derivatives containing sulfopropyl or palmythoil/carboximethyl/sulfopropyl groups and natural extracts could be potent immunomodulators with many potential applications (possible antagonists of Dectin-1, blockers of Dectin-1 cooperation with TLR-2).

Angiogenesis, the biological process by which new capillaries are formed from pre-existing vessels, is a tightly controlled and complex process involving several factors with both stimulating and inhibiting steps. In solid tumor growth, a specific clinical turning point is the transition to the vascular phase. Once it develops an intrinsic vascular network, a tumor grows indefinitely. Tumor angiogenesis depends mainly on the release by neoplasic cells of growth factors specific for endothelial cells (ECs), able to stimulate growth of the host blood vessels. The aim of this study was to analyze the apoptotic effect of some cytostatics, Vinblastine, Rapamycin and Doxorubicin, and vegetal extracts (called VOB) isolated and purified from Vitis sp., on human EA.hy926 endothelial cell line. In a proliferation assay using Crystal Violet, we demonstrated that Vinblastine and Rapamycin cytostatics have synergistic effect on endothelial cell line EA.hy926 growth inhibition. The inhibitory effects of Vinblastine and Doxorubicin were enhanced by VOB vegetal extracts. A combined treatment of cytostatics and VOB vegetal extracts resulted in a stronger antiproliferative effect of EA.hy926 endothelial cells. Results obtained regarding the apoptosis induced on EA.hy926 endothelial cells showed that each compound alone was able to induce a significant percent of apoptotic cells in a dose-dependent manner.

There are many studies demonstrating by different experimental models that non-steroidal antiinflammatory drugs (NSAIDs), also known as cyclooxygenase-2 (COX-2) inhibitors, can modulate immune response such as lymphoid cells differentiation and proliferation. There are experimental data which show that activated B cells can express mRNA COX-2, release prostaglandins (PGs) and produce immunoglobulins in PGs dependent manner. In this study, using different COX-2 inhibitors and applying personalized immunization scheme, we confirmed that it is possible to modulate in vivo antibody response against T cell dependent antigens, substantiating the importance of PGE2 and E prostanoid receptor (EP-R) in antibody generation. Our results point out the fact that we must be more careful when we apply vaccines containing T-cell dependent antigens, such as tetanus or diphteric anatoxin, to the patients under an intense antiinflammatory treatment.