Biosynthesis of silver nanoparticles using Sambucus nigra L. fruit extract for targeting cell death in oral dysplastic cells.

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 É£H2AX 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.

The impact of silver nanoparticles phytosynthesized with Viburnum opulus L. extract on the ultrastrastructure and cell death in the testis of offspring rats.

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 cell's 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.

Influence of Different Sweeteners on the Stability of Anthocyanins from Cornelian Cherry Juice.

Cornelian cherries are red fruits which can be considered as a valuable dietary source of antioxidant biologically active compounds, especially anthocyanins. The purpose of the present study was to investigate the anthocyanins degradation process in Cornelian cherry juice supplemented with different sweeteners. Four formulations of Cornelian cherry juice were prepared using different sugars (sucrose, fructose) or artificial sweeteners (aspartame and acesulfame potassium). The obtained juices were stored at three distinct temperatures (2 °C, 25 °C, and 75 °C) in order to evaluate the effects of the sweetener and storage conditions on the pigment stability. The rate constants (k) and the half time values (t1/2) of the degradation processes were determined. The highest stability was observed for the anthocyanins from the unsweetened juice stored at 2 °C (k = 0.5·10-3 h-1), while the most accelerated degradation was registered for the fructose sweetened juice stored at 75 °C (k = 91.65·10-3 h-1). The presence of the different sweeteners in the Cornelian cherry juice affects their pigment stability during storage. The highest change in the retention of anthocyanins was determined by the presence of fructose, while acesulfame potassium had the less deleterious effect.

The in vivo modulatory effects of Cornus mas extract on photodynamic therapy in experimental tumors.

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.

Effects of silver and gold nanoparticles phytosynthesized with Cornus mas extract on oral dysplastic human 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.

Effects of silver nanoparticles functionalized with Cornus mas L. extract on architecture and apoptosis in rat testicle.

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.

UV-light mediated green synthesis of silver and gold nanoparticles using Cornelian cherry fruit extract and their comparative effects in experimental inflammation.

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.

In vitro and in vivo anti-inflammatory properties of green synthesized silver nanoparticles using Viburnum opulus L. fruits extract.

A green, rapid and cost effective method for the bio-synthesis of silver nanoparticles (AgNPs), using polyphenols present in European cranberry bush fruit extracts was developed. The obtained AgNPs were characterized by ultra-violet visible spectroscopy (UV-VIS), Fourier transform - infrared spectroscopy (FT-IR), transmission electron microscopy (TEM) and X-ray diffraction patterns (XRD). The average size of the spherical AgNPs was found to be 25nm. The anti-inflammatory effect of the biomaterials was investigated, both in vitro (on HaCaT cell line, exposed to UVB radiation) and in vivo (on acute inflammation model in Wistar rats). Our results support the conclusion that the photosynthesized silver nanoparticles present a potent anti-inflammatory activity and could be successfully used as therapeutic tools for treatment of inflammation.

The effect of Sambucus nigra L. extract and phytosinthesized gold nanoparticles on diabetic rats.

Nanomaterials such as gold nanoparticles (NPs) conjugated with natural products have shown good results in lowering the glycated hemoglobin and have an anti-inflamatory effect. The aim of our study is to evaluate the antidiabetic effect of NPs functionalized with Sambucus nigra L. (SN) extract on experimental model of diabetes in rats. Diabetes was induced to 18 Wistar male rats (n=6) by a single intramuscular injection of streptozotocin (30mg/kg body weight - b.w.). SN extract (15mg/kg b.w.), NPs (0.3mg/kg b.w.) and vehicle (normal saline) were administered by gavage once a day, every morning, for 2 weeks. Other 18 animals were used as control groups and were treated with the same compounds, at the same time. Afterwards, blood, liver and muscle samples were taken to assess the oxidant/antioxidant status and the liver for the evaluation of metalloproteinases (MMP)-2 and 9 activities, COX-2 and NFKB expressions and for immunohistochemistry. Serum glycemia, cholesterol, alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT) were also measured. The administration of NPs extract increased the muscle and systemic GSH/GSSG ratio in the diabetic group vs. diabetic (p<0.03) or non-diabetic groups treated with vehicle (p<0.05) and decreased MDA levels compared to non-diabetic group (p<0.05). COX-2 expression (p<0.0001) and proMMP-2 activity (p<0.05) decreased after pretreatment with NPs in parallel with the reduction of Kupffer cells percent (<0.001). No morphological abnormalities were detected in histopathology. NPs present a great potential for further usage as adjuvants in the diabetic therapy due to the increase of antioxidant defence and reduction of MMPs activity and inflammation in liver tissue.

Green synthesis, characterization and anti-inflammatory activity of silver nanoparticles using European black elderberry fruits extract.

This research aimed at reporting the synthesis, characterization and evaluation of the anti-inflammatory effects of some new biomaterials based on silver nanoparticles and polyphenols rich natural extracts. A fast and eco-friendly extracellular biosynthesis of silver nanoparticles (AgNPs), using European black elderberry (Sambucus nigra - SN, Adoxaceae family) fruit extracts was developed. The phytosynthesized nanoparticles exhibited an absorbance peak at 426nm, characteristic for AgNPs and their sizes were ranged from 20 to 80nm. The anti-inflammatory properties of AgNPs were assessed in vitro on HaCaT cells exposed to UVB radiation, in vivo on acute inflammation model and in humans on psoriasis lesions. In vitro, our results demonstrated the anti-inflammatory effects of functionalized AgNPs by the decrease of cytokines production induced by UVB irradiation. In vivo, the pre-administration of AgNPs reduced the edema and cytokines levels in the paw tissues, early after the induction of inflammation. The present study also demonstrated the possible use of synthesized AgNPs for the treatment of psoriasis lesions.

Degradation kinetics of anthocyanins from European cranberrybush (Viburnum opulus L.) fruit extracts. Effects of temperature, pH and storage solvent.

European cranberrybush (Viburnum opulus L.) fruits are well known for their biological properties, of which some are due to the presence of anthocyanins in the berries. Current literature provides little information concerning these fruits. The stability of anthocyanins from Viburnum opulus fruits, in aqueous and ethanolic extracts, stored under darkness for 7 days at different temperatures (2 °C, 37 °C and 75 °C) and pH values (pH = 3 and 7), was studied here. The lowest stability was showed by the anthocyanins from the water extract stored at 75 °C and pH = 7, with half-life and constant rate values of 1.98 h and 0.3488 h⁻¹, respectively. The results showed a good correlation between the total anthocyanin content (determined using the pH differential method) and the time of storage, with determination coefficients varying from R² = 0.9298 to R² = 0.9971. Results indicate that the storage degradation of anthocyanins followed first-order reaction kinetics under all investigated conditions.