Elucidating the bacterial inactivation mechanism by argon cold atmospheric pressure plasma jet through spectroscopic and imaging techniques.

AIMS: This study aims to assess the potential bacterial inactivation pathway triggered by argon (Ar) cold atmospheric pressure plasma jet (CAPJ) discharge using spectroscopic and imaging techniques. METHODS AND RESULTS: Electrical and reactive species of the Ar CAPJ discharge was characterized. The chemical composition and morphology of bacteria pre- and post-CAPJ exposure were assessed using Fourier transform infrared (FTIR), Raman micro-spectroscopy, and transmission electron microscopy (TEM). A greater than 6 log reduction of Escherichia coli and Staphylococcus aureus was achieved within 60 and 120 s of CAPJ exposure, respectively. Extremely low D-values (<20 s) were recorded for both the isolates. The alterations in the FTIR spectra and Raman micro-spectra signals of post-CAPJ exposed bacteria revealed the degree of destruction at the molecular level, such as lipid peroxidation, protein oxidation, bond breakages, etc. Further, TEM images of exposed bacteria indicated the incurred damages on cell morphology by CAPJ reactive species. Also, the inactivation process varied for both isolates, as evidenced by the correlation between the inactivation curve and FTIR spectra. It was observed that the identified gas-phase reactive species, such as Ar I, O I, OH•, NO+, OH+, NO2-, NO3-, etc. played a significant role in bacterial inactivation. CONCLUSIONS: This study clearly demonstrated the effect of CAPJ exposure on bacterial cell morphology and molecular composition, illuminating potential bacterial inactivation mechanisms.

Effects of Argon Gas Plasma Treatment on Biocompatibility of Nanostructured Titanium.

In this study, we applied argon plasma treatment to titanium surfaces with nanostructures deposited by concentrated alkali treatment and investigated the effects on the surface of the material and the tissue surrounding an implant site. The results showed that the treatment with argon plasma removed carbon contaminants and increased the surface energy of the material while the nanoscale network structure deposited on the titanium surface remained in place. Reactive oxygen species reduced the oxidative stress of bone marrow cells on the treated titanium surface, creating a favorable environment for cell proliferation. Good results were observed in vitro evaluations using rat bone marrow cells. The group treated with argon plasma exhibited the highest apatite formation in experiments using simulated body fluids. The results of in vivo evaluation using rat femurs revealed that the treatment improved the amount of new bone formation around an implant. Thus, the results demonstrate that argon plasma treatment enhances the ability of nanostructured titanium surfaces to induce hard tissue differentiation and supports new bone formation around an implant site.

Chemiluminescent Analysis of Oxidative Metabolism in Rat Blood under the Influence of Argon and Helium.

We studied the nature of the action of course treatment with argon and helium (1 min, 3 procedures) on the oxidative metabolism in rat blood plasma. The study was performed on 30 Wistar rats divided into 3 groups (n=10 in each group): intact and 2 experimental (treatment of the skin of the back with a stream of argon and helium, respectively). After completion of the treatment course, the intensity of free radical processes, the total antioxidant activity, and malondialdehyde concentration were evaluated in the blood plasma. It was found that argon and helium gas flows provide stimulation of antioxidant systems, but the mechanisms of their effect were different. Treatment with helium did not affect the intensity of free radical processes, but significantly increased the overall antioxidant activity of blood plasma and reduced malondialdehyde concentration in comparison with the effect of argon flow.

Positive Neuroprotective Effect of Argon Inhalation after Photochemically Induced Ischemic Stroke Model in Rats.

We studied the effect of 2-h inhalation of argon-oxygen mixture (Ar 70%/O2 30%) after photochemically induced stroke and on days 2 and 3 after stroke modeling on the severity of neurological deficit and brain damage (by MRI data) in Wistar rats. Neurological deficit was assessed within 14 days using the limb placement test. MRI and histological study of the brain with an assessment of the size of damage were performed on day 14 after ischemia. Significant differences were obtained in limb placement scores on days 3, 7, and 14, as well as in the volume of ischemic focus by MRI in comparison with the control (ischemia+N2 70%/O2 30%). Inhalation of argon-oxygen mixture for 2 h a day over 3 days after photoinduced stroke decreased the volume of brain damage by 2 times and reduced the severity of neurological deficit.

Neuroprotection by the noble gases argon and xenon as treatments for acquired brain injury: a preclinical systematic review and meta-analysis.

BACKGROUND: The noble gases argon and xenon are potential novel neuroprotective treatments for acquired brain injuries. Xenon has already undergone early-stage clinical trials in the treatment of ischaemic brain injuries, with mixed results. Argon has yet to progress to clinical trials as a treatment for brain injury. Here, we aim to synthesise the results of preclinical studies evaluating argon and xenon as neuroprotective therapies for brain injuries. METHODS: After a systematic review of the MEDLINE and Embase databases, we carried out a pairwise and stratified meta-analysis. Heterogeneity was examined by subgroup analysis, funnel plot asymmetry, and Egger's regression. RESULTS: A total of 32 studies were identified, 14 for argon and 18 for xenon, involving measurements from 1384 animals, including murine, rat, and porcine models. Brain injury models included ischaemic brain injury after cardiac arrest (CA), neurological injury after cardiopulmonary bypass (CPB), traumatic brain injury (TBI), and ischaemic stroke. Both argon and xenon had significant (P<0.001), positive neuroprotective effect sizes. The overall effect size for argon (CA, TBI, stroke) was 18.1% (95% confidence interval [CI], 8.1-28.1%), and for xenon (CA, TBI, stroke) was 34.1% (95% CI, 24.7-43.6%). Including the CPB model, only present for xenon, the xenon effect size (CPB, CA, TBI, stroke) was 27.4% (95% CI, 11.5-43.3%). Xenon, both with and without the CPB model, was significantly (P<0.001) more protective than argon. CONCLUSIONS: These findings provide evidence to support the use of xenon and argon as neuroprotective treatments for acquired brain injuries. Current evidence suggests that xenon is more efficacious than argon overall.

Antibacterial and anticandidal effects of atmospheric-pressure, non-thermal, nitrogen- and argon-plasma pulses.

Atmospheric-pressure, non-thermal plasma destroys microorganisms by directly reacting with hydrocarbon molecules in the cell wall and/or by damaging the cytoplasmic membrane, proteins, and DNA with charged particles and reactive species. The aim of our study was to evaluate the antibacterial and anticandidal effects of atmospheric-pressure, non-thermal, nitrogen- and argon-plasma pulses on various pathogen preparations. The resultant antibacterial and anticandidal effects were assessed by evaluating percent and log reduction values for pathogen colonies. Nitrogen-plasma pulses emitted at an energy of 1.5 J and argon-plasma pulses generated at 0.5 J elicited remarkable antibacterial effects on Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA) and anticandidal effects on Candida albicans. Nitrogen-plasma pulses at a pulse count of five elicited remarkable antibacterial effects on Cutibacterium acnes at the energy settings of 1.75, 2.5, and 3 J, but not at 1 J. Meanwhile, argon-plasma pulses showed antibacterial effects on C. acnes at an energy of 0.5 and 0.65 J. Nitrogen- or argon-plasma pulses exert antibacterial and anticandidal effects on bacterial and fungal pathogens.

Inactivation of Escherichia coli by atmospheric pressure plasma jet in water.

The main aim of this work is inactivation of Escherichia coli in water using a laboratory-scale radio-frequency atmospheric pressure Argon plasma jet. This bacterium is widely present in the environment, especially in drinking water, and its pathogenic effects are very harmful. For this purpose, an Argon flow rate of 3.5 slm, maximum plasma power of 200 W, and discharge frequency of 13.56 MHz was conducted to generate a uniform plasma plume for water treatment. 150 ml of drinking water contaminated by E. coli was exposed to the radiation of plasma placed about 3 cm within the water, the treatment time varied from 2 to 6 minutes at 100, 150, and 200 W of plasma input power. The temperature of the plume, discharge current and voltage, and electron density were all measured to characterize the plasma. Active species such as excited molecules, ions, and radicals produced in the plasma in water were detected using the optical emission spectroscopy method. The decreasing behavior of live bacteria versus exposure time and plasma jet input power was observed, and finally, at the discharge power of 200 W and 6 min, an effective inactivation was achieved and the number of bacteria reduced from 92×104 to less than 1.7 MPN/100 ml.

Timely and Appropriate Administration of Inhaled Argon Provides Better Outcomes for tMCAO Mice: A Controlled, Randomized, and Double-Blind Animal Study.

BACKGROUND: Inhaled argon (iAr) has shown promising therapeutic efficacy for acute ischemic stroke and has exhibited impressive advantages over other inert gases as a neuroprotective agent. However, the optimal dose, duration, and time point of iAr for acute ischemic stroke are unknown. Here, we explored variable iAr schedules and evaluated the neuroprotective effects of acute iAr administration on lesion volume, brain edema, and neurological function in a mouse model of cerebral ischemic/reperfusion injury. METHODS: Adult ICR (Institute of Cancer Research) mice were randomly subjected to sham, moderate (1.5 h), or severe (3 h) transient middle cerebral artery occlusion (tMCAO). One hour after tMCAO, the mice were randomized to variable iAr protocols or air. General and focal deficit scores were assessed during double-blind treatment. Infarct volume, overall recovery, and brain edema were analyzed 24 h after cerebral ischemic/reperfusion injury. RESULTS: Compared with those in the tMCAO-only group, lesion volume (p < 0.0001) and neurologic outcome (general, p < 0.0001; focal, p < 0.0001) were significantly improved in the group administered iAr 1 h after stroke onset (during ischemia). Short-term argon treatment (1 or 3 h) significantly improved the infarct volume (1 vs. 24 h, p < 0.0001; 3 vs. 24 h, p < 0.0001) compared with argon inhalation for 24 h. The concentration of iAr was confirmed to be a key factor in improving focal neurological outcomes relative to that in the tMCAO group, with higher concentrations of iAr showing better effects. Additionally, even though ischemia research has shown an increase in cerebral damage proportional to the ischemia time, argon administration showed significant neuroprotective effects on infarct volume (p < 0.0001), neurological deficits (general, p < 0.0001; focal, p < 0.0001), weight recovery (p < 0.0001), and edema (p < 0.0001) in general, particularly in moderate stroke. CONCLUSIONS: Timely iAr administration during ischemia showed optimal neurological outcomes and minimal infarct volumes. Moreover, an appropriate duration of argon administration was important for better neuroprotective efficacy. These findings may provide vital guidance for using argon as a neuroprotective agent and moving to clinical trials in acute ischemic stroke.

Experimental Evaluation of the Effect of Argon Cold Plasma on Oxidative Metabolism of the Blood.

We studied the effect of course exposure to argon cold plasma (ten 1- and 2-min procedures) on some parameters of the oxidative metabolism of rat blood plasma. The intensity of free radical processes, the total antioxidant activity, and malondialdehyde concentration in rat plasma were evaluated. It was found that 2-min exposure to argon cold plasma and nonionized argon stream produce a prooxidant effect, while 1-min exposure argon plasma led to stimulation of the antioxidant reserves of the blood.

The amino acid metabolism is essential for evading physical plasma-induced tumour cell death.

BACKGROUND: Recent studies have emphasised the important role of amino acids in cancer metabolism. Cold physical plasma is an evolving technology employed to target tumour cells by introducing reactive oxygen species (ROS). However, limited understanding is available on the role of metabolic reprogramming in tumour cells fostering or reducing plasma-induced cancer cell death. METHODS: The utilisation and impact of major metabolic substrates of fatty acid, amino acid and TCA pathways were investigated in several tumour cell lines following plasma exposure by qPCR, immunoblotting and cell death analysis. RESULTS: Metabolic substrates were utilised in Panc-1 and HeLa but not in OVCAR3 and SK-MEL-28 cells following plasma treatment. Among the key genes governing these pathways, ASCT2 and SLC3A2 were consistently upregulated in Panc-1, Miapaca2GR, HeLa and MeWo cells. siRNA-mediated knockdown of ASCT2, glutamine depletion and pharmacological inhibition with V9302 sensitised HeLa cells to the plasma-induced cell death. Exogenous supplementation of glutamine, valine or tyrosine led to improved metabolism and viability of tumour cells following plasma treatment. CONCLUSION: These data suggest the amino acid influx driving metabolic reprogramming in tumour cells exposed to physical plasma, governing the extent of cell death. This pathway could be targeted in combination with existing anti-tumour agents.

Inhaled gases as novel neuroprotective therapies in the postcardiac arrest period.

PURPOSE OF REVIEW: The purpose of this review is to summarize recent advances about inhaled gases as novel neuroprotective agents in the postcardiac arrest period. RECENT FINDINGS: Inhaled gases, as nitric oxide (NO) and molecular hydrogen (H2), and noble gases as xenon (Xe) and argon (Ar) have shown neuroprotective properties after resuscitation. In experimental settings, the protective effect of these gases has been demonstrated in both in-vitro studies and animal models of cardiac arrest. They attenuate neuronal degeneration and improve neurological function after resuscitation acting on different pathophysiological pathways. Safety of both Xe and H2 after cardiac arrest has been reported in phase 1 clinical trials. A randomized phase 2 clinical trial showed the neuroprotective effects of Xe, combined with targeted temperature management. Xe inhalation for 24 h after resuscitation preserves white matter integrity as measured by fractional anisotropy of diffusion tensor MRI. SUMMARY: Inhaled gases, as Xe, Ar, NO, and H2 have consistently shown neuroprotective effects in experimental studies. Ventilation with these gases appears to be well tolerated in pigs and in preliminary human trials. Results from phase 2 and 3 clinical trials are needed to assess their efficacy in the treatment of postcardiac arrest brain injury.

Influence of helium, xenon, and other noble gases on cryopreservation of Hela and l929 cell lines.

Any biological material contains dissolved gases that affect physical and biological processes associated with cooling and freezing. However, in the cryobiology literature, little attention has been paid to the effect of gasses on cryopreservation. We studied the influence of helium, neon, krypton, xenon, argon, nitrogen, and sulfur hexafluoride on the survivability of HeLa and L929 cell lines during cryopreservation. Saturation of a cell suspension with helium, neon, and sulfur hexafluoride enhanced survival of HeLa and L929 cells after cryopreservation. Helium exerted the most significant effect. For a range of noble gases, the efficiency of the positive effect decreased as the molecular mass of the gas increased. This paper discusses possible mechanisms for the influence of gases on the cryopreservation of biological material. The most probable mechanism is the disruption of the frozen solution structure with gas-filled microbubbles produced during water crystallization. Ultimately, it was concluded that helium and neon can be used to improve methods for cryopreservation of cell suspensions with a low concentration of conventional penetrating cryoprotectants or even without them.

Argon Inhalation for 24 h After Closed-Head Injury Does not Improve Recovery, Neuroinflammation, or Neurologic Outcome in Mice.

BACKGROUND/OBJECTIVE: In recent years, the noble gas argon (Ar) has been extensively studied for its organ protection properties. While mounting in vitro and in vivo evidence indicates that argon provides neuroprotection in ischemic brain injury, its neuroprotective potential in traumatic brain injury (TBI) has not been evaluated in vivo. We tested the hypothesis that prolonged inhalation of 70% or 79% argon for 24 h after closed-head injury (CHI) improves neurologic outcome and overall recovery at 36 days post-injury. We also compared effects of the 30% or 21% residual oxygen on argon's potential neuroprotective capacity. METHODS: Adult male C57/black mice (n = 240) were subjected to closed-head traumatic brain injury, followed by inhalation of 70% argon or nitrogen (30% oxygen), or 79% argon or nitrogen (21% oxygen) for 24 h. Neurologic outcome (rotarod, neuroscore, and Morris water maze) was evaluated for up to 36 days post-injury. Histologic parameters of neurologic degeneration (Fluoro-Jade staining) and inflammation (F4/80 microglia immunostaining) were assessed in subgroups at 24 h and on post-injury day 7. RESULTS: Our CHI protocol consistently resulted in significant brain injury. After argon inhalation for 24 h at either concentration, mice did not show significant improvement with regard to neuroscores, rotarod performance, Morris water maze performance, or overall recovery (body weight), compared to nitrogen controls, up to 36 days. At 7 days post-injury, histologic markers of neurodegeneration and inflammation, particularly in the hippocampus, consistently demonstrated significant injury. Notably, recovery was reduced in mice treated with the higher oxygen concentration (30%) after CHI compared to 21%. CONCLUSIONS: Prolonged argon treatment did not improve neurologic outcome, overall recovery (weight), nor markers of neurodegeneration or neuroinflammation after significant CHI compared to nitrogen. While neuroprotective in predominately ischemic injury, argon did not provide protection after TBI in this model, highlighting the crucial importance of assessing argon's strengths and weaknesses in preclinical models to fully understand its organ protective potential in different pathologies and gas mixtures.

Argon Plasma Exposure Augments Costimulatory Ligands and Cytokine Release in Human Monocyte-Derived Dendritic Cells.

Cold physical plasma is a partially ionized gas expelling many reactive oxygen and nitrogen species (ROS/RNS). Several plasma devices have been licensed for medical use in dermatology, and recent experimental studies suggest their putative role in cancer treatment. In cancer therapies with an immunological dimension, successful antigen presentation and inflammation modulation is a key hallmark to elicit antitumor immunity. Dendritic cells (DCs) are critical for this task. However, the inflammatory consequences of DCs following plasma exposure are unknown. To this end, human monocyte-derived DCs (moDCs) were expanded from isolated human primary monocytes; exposed to plasma; and their metabolic activity, surface marker expression, and cytokine profiles were analyzed. As controls, hydrogen peroxide, hypochlorous acid, and peroxynitrite were used. Among all types of ROS/RNS-mediated treatments, plasma exposure exerted the most notable increase of activation markers at 24 h such as CD25, CD40, and CD83 known to be crucial for T cell costimulation. Moreover, the treatments increased interleukin (IL)-1α, IL-6, and IL-23. Altogether, this study suggests plasma treatment augmenting costimulatory ligand and cytokine expression in human moDCs, which might exert beneficial effects in the tumor microenvironment.

Effect of Argon-Based Atmospheric Pressure Plasma Treatment on Hard Tissue Formation on Titanium Surface.

In this paper, we suggest that the atmospheric pressure plasma treatment of pure titanium metal may be useful for improving the ability of rat bone marrow cells (RBMCs) to induce hard tissue differentiation. Previous studies have reported that the use of argon gas induces a higher degree of hard tissue formation. Therefore, this study compares the effects of plasma treatment with argon gas on the initial adhesion ability and hard tissue differentiation-inducing ability of RBMCs. A commercially available titanium metal plate was used as the experimental material. A plate polished using water-resistant abrasive paper #1500 was used as the control, and a plate irradiated with argon mixed with atmospheric pressure plasma was used as the experimental plate. No structural change was observed on the surface of the titanium metal plate in the scanning electron microscopy results, and no change in the surface roughness was observed via scanning probe microscopy. X-ray photoelectron spectroscopy showed a decrease in the carbon peak and the formation of hydroxide in the experimental group. In the distilled water drop test, a significant decrease in the contact angle was observed for the experimental group, and the results indicated superhydrophilicity. Furthermore, the bovine serum albumin adsorption, initial adhesion of RBMCs, alkaline phosphatase activity, calcium deposition, and genetic marker expression of rat bone marrow cells were higher in the experimental group than those in the control group at all time points. Rat distal femur model are used as in vivo model. Additionally, microcomputed tomography analysis showed significantly higher results for the experimental group, indicating a large amount of the formed hard tissue. Histopathological evaluation also confirmed the presence of a prominent newly formed bone seen in the images of the experimental group. These results indicate that the atmospheric pressure plasma treatment with argon gas imparts superhydrophilicity, without changing the properties of the pure titanium plate surface. It was also clarified that it affects the initial adhesion of bone marrow cells and the induction of hard tissue differentiation.

Patient-Derived Human Basal and Cutaneous Squamous Cell Carcinoma Tissues Display Apoptosis and Immunomodulation following Gas Plasma Exposure with a Certified Argon Jet.

Reactive oxygen species (ROS) have been subject of increasing interest in the pathophysiology and therapy of cancers in recent years. In skin cancer, ROS are involved in UV-induced tumorigenesis and its targeted treatment via, e.g., photodynamic therapy. Another recent technology for topical ROS generation is cold physical plasma, a partially ionized gas expelling dozens of reactive species onto its treatment target. Gas plasma technology is accredited for its wound-healing abilities in Europe, and current clinical evidence suggests that it may have beneficial effects against actinic keratosis. Since the concept of hormesis dictates that low ROS levels perform signaling functions, while high ROS levels cause damage, we investigated herein the antitumor activity of gas plasma in non-melanoma skin cancer. In vitro, gas plasma exposure diminished the metabolic activity, preferentially in squamous cell carcinoma cell (SCC) lines compared to non-malignant HaCaT cells. In patient-derived basal cell carcinoma (BCC) and SCC samples treated with gas plasma ex vivo, increased apoptosis was found in both cancer types. Moreover, the immunomodulatory actions of gas plasma treatment were found affecting, e.g., the expression of CD86 and the number of regulatory T-cells. The supernatants of these ex vivo cultured tumors were quantitatively screened for cytokines, chemokines, and growth factors, identifying CCL5 and GM-CSF, molecules associated with skin cancer metastasis, to be markedly decreased. These findings suggest gas plasma treatment to be an interesting future technology for non-melanoma skin cancer topical therapy.

Argon Attenuates Multiorgan Failure in Relation with HMGB1 Inhibition.

Argon inhalation attenuates multiorgan failure (MOF) after experimental ischemic injury. We hypothesized that this protection could involve decreased High Mobility Group Box 1 (HMGB1) systemic release. We investigated this issue in an animal model of MOF induced by aortic cross-clamping. Anesthetized rabbits were submitted to supra-coeliac aortic cross-clamping for 30 min, followed by 300 min of reperfusion. They were randomly divided into three groups (n = 7/group). The Control group inhaled nitrogen (70%) and oxygen (30%). The Argon group was exposed to a mixture of argon (70%) and oxygen (30%). The last group inhaled nitrogen/oxygen (70/30%) with an administration of the HMGB1 inhibitor glycyrrhizin (4 mg/kg i.v.) 5 min before aortic unclamping. At the end of follow-up, cardiac output was significantly higher in Argon and Glycyrrhizin vs. Control (60 ± 4 and 49 ± 4 vs. 33 ± 8 mL/kg/min, respectively). Metabolic acidosis was attenuated in Argon and Glycyrrhizin vs. Control, along with reduced amount of norepinephrine to reverse arterial hypotension. This was associated with reduced interleukin-6 and HMGB1 plasma concentration in Argon and Glycyrrhizin vs. Control. End-organ damages were also attenuated in the liver and kidney in Argon and Glycyrrhizin vs. Control, respectively. Argon inhalation reduced HMGB1 blood level after experimental aortic cross-clamping and provided similar benefits to direct HMGB1 inhibition.

Low-Temperature Argon Plasma Regulates Skin Moisturizing and Melanogenesis-Regulating Markers through Yes-Associated Protein.

Extensive water loss and melanin hyperproduction can cause various skin disorders. Low-temperature argon plasma (LTAP) has shown the possibility of being used for the treatment of various skin diseases, such as atopic dermatitis and skin cancer. However, the role of LTAP in regulating skin moisturizing and melanogenesis has not been investigated. In this study, we aimed to determine the effect of LTAP on yes-associated protein (YAP), a major transcriptional coactivator in the Hippo signaling pathway that is involved in skin moisturizing and melanogenesis-regulating markers. In normal human epidermal keratinocytes (NHEKs), the human epidermal keratinocyte line HaCaT, and human dermal fibroblasts (HDFs), we found that LTAP exhibited increased expression levels of YAP protein. In addition, the expression levels of filaggrin (FLG), which is involved in natural moisturizing factors (NMFs), and hyaluronic acid synthase (HAS), transglutaminase (TGM), and involucrin (IVL), which regulate skin barrier and moisturizing, were also increased after exposure to LTAP. Furthermore, collagen type I alpha 1 and type III alpha 1 (COL1A1, COL3A1) were increased after LTAP exposure, but the expression level of matrix metalloproteinase-3 (MMP-3) was reduced. Moreover, LTAP was found to suppress alpha-melanocyte stimulating hormone (α-MSH)-induced melanogenesis in murine melanoma B16F10 cells and normal human melanocytes (NHEMs). LTAP regulates melanogenesis of the melanocytes through decreased YAP pathway activation in a melanocortin 1 receptor (MC1R)-dependent manner. Taken together, our data show that LTAP regulates skin moisturizing and melanogenesis through modulation of the YAP pathway, and the effect of LTAP on the expression level of YAP varies from cell to cell. Thus, LTAP might be developed as a treatment method to improve the skin barrier, moisture content, and wrinkle formation, and to reduce melanin generation.

Photoautotrophic cultures of Chlamydomonas reinhardtii: sulfur deficiency, anoxia, and hydrogen production.

The aim of this work was a comparative study of S-repleted and S-depleted photoautotrophic cultures of Chlamydomonas reinhardtii under aerobic and anoxic conditions with the main focus on PSII activity. For that we used photobioreactor with short light path connected on-line to PAM fluorometer and cultivated microalgae in twice concentrated HS medium to avoid any uncontrolled limitation by mineral elements. Photoautotrophic cultures grown under Ar + CO2 gas mixture did not reach the same Chl (a + b) concentration as control culture (grown under air + CO2). At pO2 40% of air saturation (96 µM O2), the actual quantum yield of PSII started to decrease. Under microaerobic conditions when cultures stopped growing, the most significant changes in PSII function were observed. Maximum quantum yield Fv/Fm decreased significantly along with performance index, PIabs. It was accompanied by increase of fluorescence at J point, Vj. Results indicate that microaerobic conditions are stressful for photoautotrophic cultures. Photoautotrophic cultures of microalgae under S-deprivation in aerobic or anaerobic conditions showed similar behavior as photoheterotrophic ones described earlier. However, photoautotrophic cultures during anaerobiosis establishment did not show sharp "switch off" effect of actual quantum yield. We show also that S-deprivation under air or argon as well as the growth under Ar + CO2 cause significant increase of initial rise of fluorescence, which indicates that PSII and oxygen-evolving complex might be disintegrated.

Fibroblast Interaction with Different Abutment Surfaces: In Vitro Study.

BACKGROUND: Attaining an effective mucosal attachment to the transmucosal part of the implant could protect the peri-implant bone. AIM: To evaluate if chair side surface treatments (plasma of Argon and ultraviolet light) may affect fibroblast adhesion on different titanium surfaces designed for soft tissue healing. METHODS: Grade 5 titanium discs with four different surface topographies were subdivided into 3 groups: argon-plasma; ultraviolet light, and no treatment. Cell morphology and adhesion tests were performed at 20 min, 24 h, and 72 h. RESULTS: Qualitative observation of the surfaces performed at the SEM was in accordance with the anticipated features. Roughness values ranged from smooth (MAC Sa = 0.2) to very rough (XA Sa = 21). At 20 min, all the untreated surfaces presented hemispherical cells with reduced filopodia, while the cells on treated samples were more spread with broad lamellipodia. However, these differences in spreading behavior disappeared at 24 h and 72 h. Argon-plasma, but not UV, significantly increased the number of fibroblasts independently of the surface type but only at 20 min. Statistically, there was no surface in combination with a treatment that favored a greater cellular adhesion. CONCLUSIONS: Data showed potential biological benefits of treating implant abutment surfaces with the plasma of argon in relation to early-stage cell adhesion.