Effects of Photobiomodulation with Low-level Laser Therapy on Muscle Repair Following a Peripheral Nerve Injury in Wistar Rats

Peripheral nerve injury (PNI) can lead to sensory and/or motor impairment. As a treatment photobiomodulation (PBM) has demonstrated positive effects in terms of the maintenance of muscle activation and trophism. Wistar rats were divided into five groups: control, injury, injury + PBMn (irradiation over injured nerve), injury + PBMm (irradiation over affected muscle) and injury + PBMnm (irradiation over nerve and muscle). The left sciatic nerve was submitted to a crushing injury. Treatment was administered with low-level laser (780 nm, 0.04 cm², 1 W cm⁻², 3.2 J) over the injured nerve and/or the tibialis anterior muscle. The effects of PBM were favorable on muscle morphology and gene expression of calcineurin, myogenin and acetylcholine receptors. PBM led to an acceleration on muscle repair process, and effects were more evident in 2 weeks after PNI. Thus, PBM is indicated for the area over both the injured nerve and the affected muscle.     

Effects of a Single Exposure to UVB Radiation on the Activities and Protein Levels of Copper-Zinc and Manganese Superoxide Dismutase in Cultured Human Keratinocytes

Abstract— Ultraviolet B irradiation has been believed to decrease or impair the activity of reactive oxygen species (ROS) scavenging enzymes such as superoxide dismutase (SOD) in the skin. It has been recently reported that two isozymes of SOD, namely copper-zinc SOD (Cu-Zn SOD) and manganese SOD (Mn SOD), exist in mammalian cells and that the two enzymes play different roles in living systems. The aim of this study was to investigate changes in SOD activities and protein levels in cultured human keratinocytes after acute UVB irradiation. In addition, the protein levels of Cu-Zn SOD and Mn SOD were quantified separately. A single exposure to UVB irradiation produced an increase in SOD activity and protein level that peaked immediately after UVB irradiation, after which a decline was observed, with subsequent recovery to baseline levels 24 h after irradiation. In individual assays of Mn SOD and Cu-Zn SOD, the amount of Mn SOD protein decreased and then gradually recovered 24 h after irradiation. In contrast, the amount of Cu-Zn SOD protein increased immediately after UVB irradiation, and then gradually declined. To evaluate the mechanisms of these changes, we examined the effects of the cytokines, interleukin-1α (IL-1α) and tumor necrosis factor-α (TNF-α), which can be secreted from keratinocytes after UVB irradiation, on the SOD activity and protein levels in keratinocytes. Interleukin-la and TNF-α enhanced both the SOD activity and protein level of Mn SOD, while these cytokines had no effect on Cu-Zn SOD protein levels in cultured human keratinocytes after incubation for 24 h. Furthermore, when neutralizing antibodies against IL-1α and TNF-α were added separately or together to the culture medium before UVB irradiation, the recovery of total SOD activity and Mn SOD protein level were markedly inhibited 24 h after irradiation. Our results suggest that significant increases in SOD activity and protein level occur as a cutaneous antioxidant defense mechanism that protects against the cytotoxicity as a result of UVB irradiation, and that this increase in SOD is attributed to Cu-Zn SOD. The Cu-Zn SOD and Mn SOD protein levels changed in a different manner after UVB irradiation. The former may participate in an early phase and the latter in a late phase defense mechanism directed against oxidant cytotoxicity through UVB irradiation. In addition, the recovery of Mn SOD to baseline levels 24 h after UVB irradiation seems to be mediated through cytokines such as IL-1α and TNF-α, which are secreted from keratinocytes.     

Activity-guided investigation of Carissa carandas (L.) roots for anti-inflammatory constituents

The present study was structured to investigate the anti-inflammatory potential of the extracts, fractions and compounds isolated from Carissa carandas (L.) roots. Bioassay guided fractionation of methanol extract based on inhibitory potential towards proinflammatory mediators (TNF-α, IL-1β and nitric oxide (NO)) led to the identification of stigmasterol (1), lupeol (2), oleanolic acid (3), carissone (4) and scopoletin (5) as potential anti-inflammatory agents. Carissone (4) (IC₅₀ = 20.1 ± 2.69 μg/mL) and scopoletin (5) (IC₅₀ = 24.6 ± 1.36 μg/mL) exhibited significant inhibition of NO production comparable to specific NO inhibitor (L-NAME; IC₅₀ = 19.82 ± 1.64 μg/mL) without affecting the cell viability. Also, 4 and 5 at a concentration of 30 μM were found to inhibit 41.88–53.44% of TNF-α and IL-1β. To the best of our knowledge, this is the first report displaying the anti-inflammatory effects of C. carandas (L.) roots, partially mediated by inhibition of TNF-α, IL-1β and NO.     

Isolation of a new natural kingiside aglucone derivative and other anti-inflammatory constituents from Syringa reticulata

Abstract

A new secoiridoid, (1R,5S,8S,9R)-1-methyl-kingiside aglucone (1), along with nine known compounds (2–10), were isolated from the ethanol extract of the stem bark of Syringa reticulata (Bl.) Hara. The structure of compound 1 was elucidated using HR-ESI-MS, 1D and 2D NMR spectroscopy. Compounds 1–10 were evaluated for their inhibitory activity against NO, TNF-α and IL-6 production. Compounds 1, 3, 5 and 7–10 significantly inhibited the levels of NO, TNF-α and IL-6 in LPS-induced RAW264.7 cells from concentrations of 3 to 30?μM.     

Potential cardioprotective influence of bupropion against CCl4-triggered cirrhotic cardiomyopathy

Bupropion, an atypical anti-depressant and smoking cessation aid, attenuates complications arising from the activation of inflammatory and oxidative pathways. In this study, the effect of bupropion on an inflammatory and oxidative condition induced by carbon tetrachloride (CCl₄) namely cirrhotic cardiomyopathy (CCM) was investigated in rats. CCM was induced by intraperitoneal injection of CCl₄ (0.4 g/kg, i.p.). Bupropion was treated orally at doses 30 and 60 (mg/kg, p.o.) for 8 weeks. CCl₄ treatment significantly lowered hepatic antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH) while enhanced Malondialdehyde (MDA). Elevations in serum nitric oxide (NO) metabolites nitrite/nitrate, and cardiac tumor necrosis factor alpha (TNF-α) and interleukin 1-beta (IL-1β) levels were observed. Cirrhosis also decreased contractility in response to isoproterenol (10^?10 to 10^?5 M). The spleen weight and intrasplenic pressure increased and QTc, QRS and RR intervals prolonged. Pathological damages in the liver for example focal necrosis, fibrosis and the hepatic blocking increased. On the other hand, bupropion increased GSH, CAT and SOD and lowered MDA. Bupropion reduced NO metabolites and TNF-α levels and decreased IL-1β. The cardiac contractile force improved at maximal effect (Rmax) 10^-5 M by bupropion. The intrasplenic pressure was reduced by bupropion. Bupropion reduces QTc, QRS and RR intervals and the liver tissue damages. Bupropion played a cardioprotective role reducing inflammatory and oxidative factors. It may recover the impairment of cardiac contractility and hyperdynamic condition in CCM, and this effect could be mediated at least in part by a NO-dependent mechanism.     

Combination of Natural Extracts and Photobiomodulation in Keratinocytes Subjected to UVA Radiation

Natural extracts (NE) with antioxidant properties can minimize the effects of photoaging. Photobiomodulation (PBM) has proven to be a useful tool for the modulation of cell metabolism. Here, we investigate the associations of antioxidants with PBM with the aim of promoting skin rejuvenation. We began with standardization of the experimental protocol. Extracts of chamomile, rosemary, blueberry, green tea, figs, pomegranate and nutwood were tested. A custom irradiation system (366 ± 10 nm) was used to simulate sun exposure. A light emitting diode system (640 ± 12.5 nm) was used for PBM. Viability assessments were performed by the (3‐(4,5‐Dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide) MTT assay method. Based on the results, radiant exposure to UVA was defined as 9 and 1 J cm^?2 for PBM. Extract concentrations were established on the basis of dark toxicities, which ranged from 0.01% to 0.3%. The data show that PBM is a promising therapy to restore keratinocytes after UVA damage; however, the detailed mechanism and effects require further exploration. Moreover, although the combination of PBM with NE may be a useful strategy, the choice of a NE is challenging, since the working concentration and other properties, such as photosensitivity, may bring about unwanted results.     

Synthesis and anti-inflammatory and antimicrobial activities of some novel 2-methylquinazolin-4(3H)-one derivatives bearing urea,thiourea and sulphonamide functionalities

A variety of novel 2-methylquinazolin-4(3H)-one derivatives (1–29) bearing urea, thiourea and sulphonamide functionalities at position 3 of biological interest have been synthesized and screened for their anti-inflammatory (TNF-α and IL-6) and antimicrobial activities (antibacterial and antifungal). “Biological evaluation study revealed that the compounds 3, 4, 6, 9, 16 and 18 were found to have promising anti-inflammatory activity (up to 62–84% TNF-α and 73–92% IL-6 inhibitory activity) at concentration of 10 μM with reference to standard dexamethasone (75% TNF-α and 84% IL-6 inhibitory activities at 1 μM) and 7, 10, 12, 23, 25 and 28 have antimicrobial activity at MIC of 10–30 μg/mL against selected pathogenic bacteria and fungi.”     

Evaluation of the pharmacological function of ulinastatin in experimental animals

Organ protection is a routine therapy in severe injuries. Our aim was to evaluate the beneficial effects of ulinastatin in experimental rats. Rats were randomly divided into a sham control, a model control and an ulinastatin-treated group. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were determined. Serum amylase, serum aspartate aminotransaminase (AST), lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CKMD) activities, interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), nitric oxide (NO) and cardiac troponin I (nTnl) levels were examined. Results showed that ulinastatin decreased MDA levels and ameliorated the down-regulation of SOD activity. In addition, ulinastatin pretreatment may decrease serum AST, LDH and CKMD activities, IL-8, TNF-α, and nTnl levels, and enhance NO level. Our results demonstrated that oxidative injury occurred after IR and that ulinastatin exhibits significant protective effects against these effects.     

Low Level Light Therapy Modulates Inflammatory Mediators Secreted by Human Annulus Fibrosus Cells during Intervertebral Disc Degeneration In Vitro

Intervertebral disc degeneration (IVD) is one of the important causes of low back pain and is associated with inflammation induced by interaction between macrophages and the human annulus fibrosus (AF) cells. Low‐level light therapy (LLLT) has been widely known to regulate inflammatory reaction. However, the effect of LLLT on macrophage‐mediated inflammation in the AF cells has not been studied till date. The aim of this study is to mimic the inflammatory microenvironment and to investigate the anti‐inflammatory effect of LLLT at a range of wavelengths (405, 532 and 650 nm) on the AF treated with macrophage‐like THP‐1 cells conditioned medium (MCM) containing proinflammatory cytokines and chemokines (interleukin‐1beta, tumor necrosis factor‐alpha, interleukin‐6 and 8). We observed that AF cells exposed to MCM secrete significantly higher concentrations of IL‐6, IL‐8, IL‐1β and TNF‐α. LLLT markedly inhibited secretion of IL‐6 at 405 nm in a time‐dependent manner. Level of IL‐8 was significantly decreased at all wavelengths in a time‐dependent manner. We showed that MCM can induce the inflammatory microenvironment in AF cells and LLLT selectively suppressed IL‐6 and 8 levels. The results indicate that LLLT is a potential method of IVD treatment and provide insights into further investigation of its anti‐inflammation effect on IVD.     

Lignans from Schisandra chinensis rattan stems suppresses primary Aβ1-42-induced microglia activation via NF-κB/MAPK signaling pathway

Microglia cells play important roles in neurodegenerative diseases for clearing amyloid-β and reducing the occurrence of inflammation. In this study, the neuroinflammatory effect and the mechanism of lignans from Schisandra chinensis rattan stems (rsSCH-L) were evaluated by Aβ_1-42-induced primary microglia cell model. The results have shown that rsSCH-L could reduce the levels of pro-inflammatory cytokines, including IL-1β, TNF-α and NO. Moreover, rsSCH-L suppressed the phosphorylations of NF-κB and IκBα as well as p38, JNK and ERK proteins in Aβ_1-42-induced microglia cells. Taken together, rsSCH-L could attenuate microglia cells from neuroinflammation by activating the NF-κB/MAPK signaling pathway.     

Modulation effect of Semen Ziziphi Spinosae extracts on IL-1β, IL-4, IL-6, IL-10, TNF-α and IFN-γ in mouse serum

This study aims to explore and evaluate the effects of Semen Ziziphi Spinosae extracts on the serum levels of interleukin (IL)-6, IFN-γ, IL-1β, TNF-α, IL-10 and IL-4 in mice, and the regulative effect of Semen Ziziphi Spinosae on the cytokine system. Using an ELISA assay, the serum levels of IL-6, IFN-γ, IL-1β, TNF-α, IL-10 and IL-4 were examined in mice after intraperitoneal injection (i.p.) with Semen Ziziphi Spinosae extracts. The results showed that the levels of IL-6 and IL-1β were significantly increased compared with the control groups (p<0.01), while the level of tumour necrosis factor-α (TNF-α) was significantly decreased (p<0.01). Semen Ziziphi Spinosae possesses certain modulation effects on cytokines, and the immuno-regulating function and hypnosis effects of Semen Ziziphi Spinosae may be relevant to these modulation effects on cytokines.     

Effects of Photobiomodulation on Daphnia magna Straus and their Sensitivity to Toxicant

This paper deals with the effect of photobiomodulation (PBM) on Daphnia magna S. and their sensitivity to cadmium sulfate, a known high toxic pollutant. In a first series of experiments, the effect of different He-Ne laser fluences irradiation (range 0.9-4300 mJ cm⁻²) on the fertility of both parent and filial generations (F1–F3) of the crustacean was studied. It was found that PBM in some cases significantly influenced the fertility of both irradiated crustaceans and their nonirradiated offspring. By selecting two fluences (9 ± 2 mJ cm⁻² reducing fertility and 4.3 ± 0.9 J cm⁻² increasing it), the effect of these on toxicity of cadmium sulfate was evaluated. These experiments have shown that prior irradiation with low-intensity light of a helium–neon laser with 632.8 nm wavelength can change the sensitivity of aquatic organisms to toxin cadmium sulfate. The degree and direction of changes depend on the toxicant concentration and the irradiation dose.     

Effects of Photobiomodulation on Root Resorption Induced by Orthodontic Tooth Movement and RANKL/OPG Expression in Rats

The aim of this study was to evaluate the effects of photobiomodulation on the repair of induced root resorption (RR) after orthodontic tooth movement. Twenty male rats were used in this study. Forty right and left upper first molars were evaluated and divided into four groups (n = 10): negative control group (NC), no tooth movement or irradiation; positive control group (PC), induced tooth movement and root resorption; conventional treatment group (CT), force was removed after 7 days; and photobiomodulation group (PBM) after force removal molars were irradiated every 48 h for 7 days using GaAlAs diode laser (810 nm). Energy per point was 1.5 J (100 mW, 15 s, 75 J cm^?2). NC and PC were euthanized on day 7; CT and PBM on day 14. Histomorphometric and immunohistochemsitry analyses showed increase in area of root resorption in all groups (P < 0.05) compared to NC. RR lacunae were larger in CT compared to PC and PBM at the compression side of the distal root. OPG was higher in PBM group (P < 0.05). PBM group showed low expression of RANKL compared to PC and CT on the tension side. PBM can potentially affect RR progression by increasing OPG expression in the compression area and decreasing number of clastic cells in the root surface.     

Bilirubin protects grafts against nonspecific inflammation-induced injury in syngeneic intraportal islet transplantation

Nonspecific inflammatory response is the major cause for failure of islet grafts at the early phase of intraportal islet transplantation (IPIT). Bilirubin, a natural product of heme catabolism, has displayed anti-oxidative and anti-inflammatory activities. The present study has demonstrated that bilirubin protected islet grafts by inhibiting nonspecific inflammatory response in a syngeneic rat model of IPIT. The inflammation-induced cell injury was mimicked by exposing cultured rat insulinoma INS-1 cells to cytokines (IL-1β, TNF-α and IFN-γ) in in vitro assays. At appropriate lower concentrations, bilirubin significantly attenuated the reduced cell viability and enhanced cell apoptosis induced by cytokines, and protected the insulin secretory function of INS-1 cells. Diabetic inbred male Lewis rats induced by streptozotocin underwent IPIT at different islet equivalents (IEQs) (optimal dose of 1000, and suboptimal doses of 750 or 500), and bilirubin was administered to the recipients every 12 h, starting from one day before transplantation until 5 days after transplantation. Administration of bilirubin improved glucose control and enhanced glucose tolerance in diabetic recipients, and reduced the serum levels of inflammatory mediators including IL-1β, TNF-α, soluble intercellular adhesion molecule 1, monocyte chemoattractant protein-1 and NO, and inhibited the infiltration of Kupffer cells into the islet grafts, and restored insulin-producing ability of transplanted islets.     

Tilianin alleviates airway inflammation in ovalbumin-induced allergic asthma in mice through the regulation of Th2 cytokines and TGF-β1/Smad markers

BackgroundAllergic asthma is a inflammatory disease defined as a condition of chronic airway inflammation. Asthma can be provoked by various stimuli like allergens inhalation like dust particles, pollen, and pollutants in the air.ObjectiveThis exploration was dedicated to investigate the anti-asthmatic properties of tilianin against the ovalbumin (OVA)-initiated asthma in mice.MethodologyThe asthma was provoked to the mice via administering 100 μl of aluminum hydroxide containing 20 μg of OVA and treated with the 10 and 20 mg/kg of tilianin, respectively. The levels of Th2 cytokines, OVA-specific IgE, eotaxin, pro-inflammatory mediators, antioxidants, and other markers were inspected by marker specific assay kits. The mRNA expressions of TGF-β1, Smad, iNOS, and COX-2 was assessed using RT-PCR analysis. The lung histology was analyzed microscopically to detect the histological changes.ResultsTilianin treatment remarkably suppressed the IL-4, IL-5, and IL-13, IFN-γ, eotaxin, and IgE levels. The NO, MPO, and inflammatory makers TNF-α, IL-6, IL-12, and TXB2 was substantially diminished by the tilianin treatment. The TGF-β1, iNOS, and COX-2 expressions were appreciably suppressed by the tilianin. The histological findings proved that the tilianin treatment alleviated the OVA-provoked histopathological changes in the lung tissues.ConclusionOur findings proved that tilianin effectively alleviated the OVA-provoked asthma in animals and it could be a talented anti-asthmatic candidate.     

Secretion of adenylate kinase 1 is required for extracellular ATP synthesis in C2C12 myotubes

Extracellular ATP (exATP) has been known to be a critical ligand regulating skeletal muscle differentiation and contractibility. ExATP synthesis was greatly increased with the high level of adenylate kinase 1 (AK1) and ATP synthase beta during C2C12 myogenesis. The exATP synthesis was abolished by the knock-down of AK1 but not by that of ATP synthase beta in C2C12 myotubes, suggesting that AK1 is required for exATP synthesis in myotubes. However, membrane-bound AK1beta was not involved in exATP synthesis because its expression level was decreased during myogenesis in spite of its localization in the lipid rafts that contain various kinds of receptors and mediate cell signal transduction, cell migration, and differentiation. Interestingly, cytoplasmic AK1 was secreted from C2C12 myotubes but not from C2C12 myoblasts. Taken together all these data, we can conclude that AK1 secretion is required for the exATP generation in myotubes.     

Effect of Ceramide on Mesenchymal Stem Cell Differentiation Toward Adipocytes

Proinflammatory cytokines such as tumor necrosis factor (TNF) α are well known to inhibit adipocyte differentiation. TNF-α triggers ceramide synthesis through binding of TNF-α to its p55 receptor. Therefore, ceramide is implicated in many of the multiple signaling pathways initiated by TNF-α. In breast tissue engineering, it is important to know how to modulate adipocyte differentiation of the stem cells with exogenous additives like ceramide in vitro. We hypothesized that stem cell adipogenesis could be retained in TNF-α-treated preadipocytes in which ceramide synthesis was blocked and that exogenous ceramide could inhibit adipocyte differentiation. We first studied the effect of ceramide synthase inhibitor, Fumonisin B2, on the adipogenesis of murine mesenchymal stem cells (D1 cells), treated with TNF-α. We then studied the effect of specific exogenous C6-ceramide on D1 cell viability and differentiation. It was found that 1 ng/ml of TNF-α significantly inhibited D1 cell adipogenesis. Cells treated with 5 μM of Fumonisin B2 were able to undergo adipogenesis, even when treated with TNF-α. High concentrations of exogenous C6-ceramide (>50 μM) had an inhibitory effect, not only on the pre-confluent proliferation of the D1 cells but also on the post-confluent cell viability. High concentrations of C6-ceramide (>50 μM) also inhibited mitotic clonal expansion when D1 cell differentiation was induced by the addition of an adipogenic hormonal cocktail. C6-ceramide at low concentrations (10–25 μM) inhibited lipid production in D1 cells, demonstrated by decreased levels of both total triglyceride content and specific fatty acid composition percentages. Genetic expression of peroxisome proliferator-activated receptor (PPAR) γ and aP2 in D1 cells was reduced by C6-ceramide treatment. CCAAT/enhancer-binding protein (C/EBP) β levels in D1 cells were reduced by C6-ceramide treatment during early differentiation; PPARγ and aP2 protein levels were reduced at terminal differentiation. C6-ceramide at lower concentrations also decreased lipid accumulation of differentiating D1 cells. Our results suggest that ceramide synthase inhibitor retains the adipogenic potential of TNF-α-treated mesenchymal stem cells, while exogenous ceramide at lower concentrations inhibit the adipogenesis of mesenchymal stem cells. Ceramide, therefore, could be a modulator candidate in breast tissue engineering strategies.     

980 nm Photobiomodulation Promotes Osteo/Odontogenic Differentiation of the Stem Cells from Human Exfoliated Deciduous Teeth Via the Cross talk Between BMP/Smad and Wnt/β-Catenin Signaling Pathways

Increasing evidence suggests stem cells from human exfoliated deciduous teeth (SHEDs) serve as desirable sources of dentin regeneration. Photobiomodulation (PBM) has shown great potential in enhancing the proliferation and osteogenesis of human bone marrow mesenchymal stem cells (hBMMSCs). However, the specific role of PBM in odontogenic differentiation of SHEDs is little know, and we further investigated potential mechanism of PBM osteo/odontogenisis. A 980 nm diode laser with different energy densities of (0.5, 5, 10 J cm⁻²) in a 100-mW continuous wave was used for irradiation every 24 h. Osteo/odontogenic differentiation of SHEDs was achieved by performing alkaline phosphatase (ALP) and alizarin red staining (ARS) and osteo/odontogenic markers were also evaluated by qRT-PCR and western blotting. Additionally, western blot and immunohistochemical staining were performed to evaluate the levels of BMP/Smad and Wnt/β-catenin signaling-related proteins. We found that PBM at 5 J cm⁻¹ increased mineral deposition and upregulated the expression of related osteo/odontogenic markers along with the elevated expression of β-catenin and phosphorylation level of Smad1/5/9. Furthermore, Wnt signaling inhibition using DKK1 and BMP signaling inhibition using noggin inhibited PBM-induced osteo/odontogenic marker expression when used individually or jointly. In conclusion, PBM induces the osteo/odontogenic differentiation of SHEDs through cross talk between BMP/Smad and Wnt/β-catenin signaling pathways.