Impact of chronic psychological stress on platelet membrane fatty acid composition in a rat model of type 1 diabetes Mellitus.

BACKGROUND: Chronic stress and diabetes mellitus are highly associated with oxidative stress and inflammation, resulting in cell membrane disruption and platelet activity. This study aims to evaluate the impact of chronic psychological stress on the composition of the platelet phospholipid membrane and platelet activation in type 1 diabetes mellitus (T1DM). METHODS: We enrolled 35 mature healthy female Wistar rats and randomly divided them into 4 groups, namely the control group (n = 9), stress group (n = 10), T1DM group (n = 8), and T1DM + Stress group (n = 8). The Wistar rats were treated in different experimental conditions for 28 days while being provided free access to feed and water. The concentration of corticosterone in blood serum and hair samples was measured using a competitive enzyme-linked immunosorbent assay. Gas chromatography-mass spectrometry was conducted to identify the methyl esters of fatty acids (FAs) in the platelet phospholipid membrane. A quantitative determination of 11-dehydro-thromboxane B2 in the blood serum was also performed using a competitive enzyme-linked immunosorbent assay. RESULTS: After 28 days, the concentration of corticosterone in blood serum (ng/mL) was observed to be higher in the stress group as compared to the T1DM and T1DM + Stress groups (P = 0.031 and P = 0.008, respectively). The percentage of C 16:0 FA in the platelet membrane was greater in the T1DM + Stress group, but its levels of C 20:1 omega (ω) 9 FA, including C 18:3ω3 FA, C 20:5ω3 FA, and the total sum of ω3 FAs, were lower as compared to the control group (P = 0.016; P = 0.016; P = 0.031; P = 0.016, P = 0.031). The concentration of 11-dehydro-thromboxane B2 in blood serum (pg/mL) was observed to be higher in the stress group than in rats with T1DM (P = 0.063). CONCLUSION: Chronic psychological stress is related to higher levels of corticosterone, saturated FAs acids in the platelet membrane, and greater platelet activation. This study proves how a low percentage of unsaturated fatty acids in the DM and stress groups indicates the disturbing impact of the oxidative/inflammatory environment to lipid metabolism and neuroendocrine response.

Assessing the efficacy of a 100 ps Nd:YAG laser for tattoo removal in a minipig model.

Using a minipig model, we evaluated the efficacy of the 100 ps Nd:YAG laser in the removal of tattoo pigments, specifically blue, green, red, and yellow. We observed distinct pigment responses to 532/1064 nm wavelengths at various energy settings. Through a combination of clinical, spectroscopic, and histological methods, we found the 532 nm wavelength to be most effective in disrupting all colors, with notable results for green and yellow at 0.4 J/cm2 and red at 0.72 J/cm2. The 1064 nm wavelength reduced pigment in yellow (1.51 J/cm2), green (1.35 J/cm2), and blue (1.11 J/cm2) tattoos, but was surpassed by the 532 nm in efficiency. Our data underscores the crucial interplay between pigment traits and laser settings in tattoo removal. We advocate for tailored treatment strategies, integrating pigment hue and laser wavelength, to enhance removal outcomes.

Comprehensive examination of tattoo removal using a 150 ps Nd:YAG laser in a porcine model.

This study aimed to investigate the efficacy of a Nd:YAG laser with a pulse duration of 150 ps at different laser parameters. The effects on multiple-colored tattoos with such ultrashort pulses has not been previously described in the literature. In vivo experiments were conducted on porcine skin to analyze the fragmentation efficiency of five different tattoo colors using different wavelengths, pulse energies, and spot sizes. The results showed that the optimal tattoo clearance to safety ratio for blue, green, red, and yellow tattoos with a 532 nm wavelength was 0.96-2.39 J/cm2. The laser with a wavelength of 1064 nm demonstrated the highest efficacy in eliminating black tattoos, with positive results observed for green and blue pigments at a fluence of 3.02 J/cm2. The study provides valuable insights into the efficacy of laser treatment with 150 ps for removing tattoos of different colors using different laser parameters. This information can help dermatologists and practitioners perform more efficient and effective tattoo removal with fewer side effects.

Microlesion healing dynamics in in vivo porcine skin after treatment with 1064 nm picosecond-domain Nd:YAG laser.

The fractionated picosecond laser produces microscopic lesions in the epidermis and dermis due to laser-induced optical breakdown (LIOB). There have been multiple histological reports, but the present literature lacks detailed in vivo studies after treatment with high-power laser systems. Our study aimed to characterize the healing patterns of microlesions induced with 150 ps duration 1064 nm MLA-type picosecond laser. The induced picosecond laser-tissue reactions with pulse energy of 50-250 mJ and different treatment modes were observed in in vivo porcine skin model over 10 days after the laser procedure. A macroscopic evaluation was combined with microscopic histological analysis to observe the healing dynamics of laser-induced microlesions. Superficial, intraepidermal cavitation bubbles were induced using microbeam fluence of 4-20 J/cm2 . Skin irritation scores positively correlated with pulse energy and dose. Our findings demonstrate that dose and pulse energy had a direct impact on epidermal thickness and lesions healing dynamics.

Antioxidant effects of gold nanoparticles on early stage of collagen-induced arthritis in rats.

OBJECTIVE: To determine antioxidant effects of prophylactic treatment with gold nanoparticles (AuNPs) in the early stage of collagen-induced arthritis (CIA). METHODS: The preventive treatment with 13 nm or 50 nm AuNPs injected intraarticularly (i.a.) was started at the induction day (0) of CIA and finished in the early stage of arthritis at the day 10. At the end of experiment blood indices (erythrocyte sedimentation rate, leukocyte and erythrocyte counts), pro-/antioxidant status of blood serum (the amount of malondialdehyde, catalase and total antioxidant activity), and internal organs' weight as well as the changes in the joint tissues and their microscopic structure were evaluated. RESULTS: Both 13 nm and 50 nm AuNPs showed antioxidant effect by increasing the level of catalase activity in the early stage of experimental arthritis. Preventive treatment with 50 nm more than with 13 nm AuNPs suppressed joint swelling. Histopathological asssesment revealed statistically significant reduction of synovial angiogenesis and erosion formation in the cartilage. Pilot transmission electron microscopy (TEM) analysis showed predominant accumulation of 13 nm in the synovial fibroblast lineage cells. CONCLUSIONS: Intraarticular injections of 13 nm or 50 nm AuNPs showed an antioxidant action significantly raising catalase activity without causing negative effects on hematological indices. Prophylactic treatment with 50 nm more than with 13 nm AuNPs suppressed joint swelling, synovial angiogenesis and cartilage erosion in the initial stage of arthritis.

The safety and efficacy of light emitting diodes-based ultraviolet A1 phototherapy in bleomycin-induced scleroderma in mice.

PURPOSE: To define the efficacy and safety of narrowband ultraviolet A1 (UVA1) for the treatment of dermal fibrosis in bleomycin-induced mouse model of scleroderma. MATERIALS AND METHODS: 42 DBA/2 strain mice were included in the study: healthy mice and mice with established scleroderma, treated with high or medium dose of UVA1. Non-treated groups served as control. The equipment emitting 365±5nm UVA1 radiation was used in the study. The average cumulative doses were 1200J/cm2 for high and 600J/cm2 for medium dose course. Histological analysis was performed for the evaluation of the dermal thickness and mast cells density. The expressions of p53 and Ki-67 proteins were assessed by immunohistochemical analyses. RESULTS: Skin thickness of mice with scleroderma, treated with high and medium dose of UVA1, were lower (272.9±113.2µm and 394±125.9µm, respectively) in comparison to the dermal thickness of non-treated animals (599±55.7µm). The dermal mast cells count in mice with scleroderma was reduced after high and medium dose treatment to 11±1.7 and 13±2.2, respectively, as compared to that in non-treated mice (23±3.0). No significant upregulation of p53 nor Ki-67 proteins was observed in the skin of healthy mice and mice with scleroderma after high- and medium-dose of UVA1. CONCLUSIONS: The results of this study indicate that 365nm UVA1 with the cumulative doses of 1200J/cm2 and 600J/cm2 is safe and effective for the dermal fibrosis treatment.

The impact of high-dose narrowband ultraviolet A1 on dermal thickness, collagen and matrix-metalloproteinases in animal model of scleroderma.

OBJECTIVE: The main purpose of the present study was to define the impact of high-dose of 365±5nm ultraviolet A1 (UVA1) on dermal fibrosis in the pre-established, bleomycin-induced mouse model of scleroderma. METHODS: DBA/2 strain mice with the pre-established, bleomycin-induced scleroderma were irradiated with cumulative UVA1 dose of 1200J/cm2 and in parallel were challenged with prolonged administration of bleomycin. Non-treated groups served as the control. Light source emitting a narrow band UVA1 light of 365±5nm and 21mW/cm2 power density was used in the study. Histological analysis was performed for the evaluation of dermal thickness. The expressions of matrix-metalloproteinase-1 (MMP-1), matrix-metalloproteinase-3 (MMP-3), collagen types I and III were evaluated by immunohistochemical analyses. The Mann - Whitney U test was used for statistical analysis. RESULTS: Dermal thickness in mice injected with bleomycin during all the experiment (8weeks) and irradiated with UVA1 for the last 5weeks was significantly lower than that in mice challenged only with bleomycin for 8weeks (253.96±31.83µm and 497.43±57.83µm, respectively; P=0.002). The dermal thickness after phototherapy was lower as compared with the pre-existing fibrotic changes observed after 3weeks of bleomycin injections (253.96±31.83µm and 443.87±41.76µm, respectively; P=0.002). High-dose of UVA1 induced the 5.8- and 5.2-fold increase in MMP-1 and MMP-3 expressions, respectively, and the 1.2- and 1.4-fold decrease in collagen type I and collagen type III expressions in the pre-established, bleomycin-induced scleroderma model as compared to that in the control non-irradiated mice (P=0.002). CONCLUSIONS: Our study has demonstrated that a cumulative 365±5nm UVA1 radiation dosage of 1200J/cm2 not only prevents the progression of dermal fibrosis, but also induces a regression of pre-existing fibrotic changes.

Cell surface markers and exogenously induced PpIX in synovial mesenchymal stem cells.

The aim of present study was to assess the expression of surface markers and the accumulation of protoporphyrin IX in synovial mesenchymal stem cells (SMSCs). SMSC from patients with rheumatoid arthritis (RA, n = 5) and osteoarthritis (OA, n = 5-6) were characterized and their PpIX accumulation rates were evaluated by flow cytometry. The expression of the 21 out of 24 tested surface markers, related to stem-like features and aggressiveness of cells showed no statistically significant differences between RA and OA groups. However, the cells from RA group had the significantly lower levels of expression for the integrin-associated protein CD47 and the grow factor receptor CD271 (P = 0.018), while the higher levels of cell membrane zinc-dependent metalloproteinase CD10 (P = 0.006), as compared to the cells from OA group. Comparison of the mean intensities of PpIX fluorescence revealed no statistically significant differences between the RA and OA groups, as well as no relation to proliferation rates or cell size, although some conspicuous distinction in PpIX accumulation was observed in certain specimens within these groups, suggesting possibilities of this method application for characterization of individual SMSC populations. CD10, CD47, and CD271 were differently expressed in RA and OA SMSC, while had no direct association with the PpIX fluorescence intensity.

Multidimensional visualization of healthy and sensitized rabbit knee tissues by means of confocal microscopy.

This study combines several fluorescence detection methods to distinguish structural features of the synovium and cartilage tissues and to visualize the localization of endogenous porphyrins in the sensitized tissues. Specimens of synovium and cartilage tissues obtained from rabbits with antigen-induced monoarthritis after intra-articular 5-aminolevulinic acid methyl ester injection and those from healthy rabbits were investigated ex vivo by means of fluorescence spectroscopy, fluorescence intensity, and lifetime microscopy. The presence of endogenous porphyrins was confirmed with the fluorescence spectra measured on sliced sensitized specimens. Application of the lifetime-gating method on fast fluorescence lifetime imaging microscopy images, allowed separate visualization of tissue structures possessing different average lifetimes. The presence of the structures has been validated by histopathological imaging based on conventional rapid hematoxylin­eosin staining of the specimens. The fluorescence lifetime of endogenous protoporphyrin IX has been assessed and employed for visualization of sensitized tissues.

Induction of protoporphyrin IX in patient-derived synoviocytes, cartilage explants and chondrons after application of 5-aminolevulinic acid or its methyl ester.

OBJECTIVE: To compare the accumulation of protoporphyrin IX between synoviocytes of patients with rheumatoid arthritis (RA) or osteoarthritis (OA) and cartilage explants (CE) as well as chondrons of patients with OA after the application of 5-aminolevulinic acid (ALA) or its methyl ester (ALA-Me). MATERIALS AND METHODS: Samples of synovial and cartilage tissues were obtained from joint replacement surgeries. The accumulation of PpIX was determined by measuring fluorescence spectra from 2 × 10(5) synoviocytes or chondrons suspended in a glass tube or directly from CE surface after 2, 4, 8 and 24h of incubation with ALA or ALA-Me. RESULTS: No differences were found between the average fluorescence intensity values of PpIX in synoviocytes of patients with RA and OA. These values were non-significantly higher after incubation with ALA in comparison with ALA-Me at almost all time points. The average fluorescence intensity of PpIX in CE and chondrons was about ten times lower than in synoviocytes. The presence of preparation of hyaluronic acid (HA) significantly enhanced PpIX induction in chondrons versus treatment only with ALA. CONCLUSIONS: A potential for the selective synovial sensitization with endogenous PpIX in comparison with cartilage tissue has been demonstrated in vitro after application of ALA or ALA-Me.