Development and Characterization of Squalene-Loaded Topical Agar-Based Emulgel Scaffold: Wound Healing Potential in Full-Thickness Burn Model.

Full-thickness burns pose a major challenge for clinicians to handle because of their restricted self-healing ability. Even though several approaches have been implemented for repairing these burnt skin tissue defects, all of them had unsatisfactory outcomes. Moreover, during recent years, skin tissue engineering techniques have emerged as a promising approach to improve skin tissue regeneration and overcome the shortcomings of the traditional approaches. Although previous literatures report the wound healing effects of the squalene oil, in the current study, for the first time, we developed a squalene-loaded emulgel-based scaffold as a novel approach for potential skin regeneration. This squalene-loaded agar-based emulgel scaffold was fabricated by using physical cross-linking technique using lecithin as an emulsifier. Characterization studies such as X-ray diffraction, Fourier-transform infrared spectroscopy, and field emission scanning electron microscopy revealed the amorphous nature, chemical interactions, and cross-linked capabilities of the developed emulgel scaffold. The squalene-loaded emulgel scaffold showed excellent wound contraction when compared with the agar gel and negative control. In case of the histopathology and recent immunohistochemistry findings, it was clearly evidenced that squalene-loaded emulgel promoted faster rate of the revascularization and macrophage polarization in order to enhance the burn wound healing. Moreover, the findings also revealed that the incorporation of squalene oil into the formulation enhances collagen deposition and accelerates the burnt skin tissue regeneration process. Finally, we conclude that the squalene-loaded emulgel scaffold could be an effective formulation used in the treatment of the burnt skin tissue defects.

Bioactive role of plant stress hormone methyl jasmonate against lipopolysaccharide induced arthritis.

The current investigation was carried out to screen antiarthritic potential of Methyl Jasmonate (MJ) against lipopolysaccharide (LPS) induced arthritis. Cartilage damage was induced in experimental animals by intraplantar administration of LPS (1 mg/kg) and antiarthritic effect of MJ was screened in two doses of MJ-1 (20 mg/kg), MJ-2 (40 mg/kg) by intraperitoneally administration. Indomethacin (30 mg/kg p.o.) was used as standard drug. The severity of arthritis was evaluated by assessing arthritis score, secondary lesions, motility test, stair climbing ability, and dorsal flexion pain score method. The estimation of blood cytokine tumor necrosis factor- aplha (TNF-α),interleukine (IL-2 and IL-6) and thymus/spleen index was carried out to access the severity of inflammation. Estimation of hepaticenzymatic antioxidant activity superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), glutathione peroxidase (GPx)and radiological examination was carried out on 28th day. Results indicated that MJ showed significant reduction in severity of arthritis by decreasing arthritis score, secondary lesions where as significant increase in motility, climbing ability and flexion pain score was observed. Significant decreased in blood cytokine viz. TNF-α, IL-2, IL-6 andthymus/spleen index was observed in MJ treated animals in dose dependent manner. MJ treated animals showed significant increased and restoration of hepatic antioxidant enzymatic activityof SOD, CAT, GSH, GPx where asradiological examination indicates protective effect on joint structure as compared to LPS treated rats. These current studies conclude that MJ has protective role in arthritis.

Molecular mechanism of plant stress hormone methyl jasmonate for its anti-inflammatory activity.

Plant stress hormones (Phytohormones/PTH) are abundantly present in numerous vascular plants. Several classes of plant stress hormones like auxins (AU) & gibberellins (GA), cytokinins (CK), abscisic acid (ABA), ethylene (ET), salicylic acid (SA), jasmonates (JA), brassinosteroids (BR) and strigolactones are synthesized within specialized plant cells. Among them, jasmonate are prominent class of stress hormones involved in survival of plants in stressful conditions. Methyl jasmonate (MeJA) is ester of jasmonic acid is extensively studied for its potential clinical benefits. MeJA is used as an effective antimicrobial agent, food preservative, antioxidant in food and agricultural sectors. The clinical benefits of MeJA have been related to their prominent interactions with inflammatory NF-κB pathways, inhibition of enzymes, gene expression for synthesis of inflammatory mediators, signaling molecules, oxidative stress and modulation of pain perception/nociceptive responses. The objective of the present review is to provide an cohesive relation of MeJA in inflammation with reference to past and recent in-vivo and in-vitro investigations in broad perspectives.

Icariin-Loaded Polyvinyl Alcohol/Agar Hydrogel: Development, Characterization, and In Vivo Evaluation in a Full-Thickness Burn Model.

Tissue regeneration has become a promising strategy for repairing damaged skin tissues. Among the hydrogels for tissue regeneration applications, topical hydrogels have demonstrated great potential for use as 3D-scaffolds in the burn wound healing process. Currently, no report has been published specifically on icariin-loaded polyvinyl alcohol (PVA)/agar hydrogel on full-thickness burn wounds. In the present study, burn tissue regeneration based on biomimetic hydrogel scaffolds was used for repairing damaged extracellular matrix. Furthermore, a skin burn model was developed in rats, and the icariin-loaded PVA/agar hydrogels were implanted into the damaged portions. The regeneration of the damaged tissues with the help of the icariin-loaded hydrogel group exhibited new translucent skin tissues and repaired extracellular matrix, indicating that the hydrogel can enhance the wound healing process. Moreover, characterization studies such as X-ray diffraction, Fourier-transformed infrared spectroscopy, and differential scanning calorimetry reported the extent of compatibility between icariin and its polymers. Results of the field emission scanning electron microscopy images revealed the extent of the spread of icariin within the polymer-based hydrogel. Furthermore, the wound healing potential, confirmed by histopathological and histochemical findings at the end of 21 days, revealed the visual evidence for the biomimetic property of icariin-loaded PVA/agar hydrogel scaffolds with the extracellular matrix for tissue regeneration.

Methyl jasmonate a stress phytohormone attenuates LPS induced in vivo and in vitro arthritis.

The objective of present study was to screen the effect of methyl jasmonate (MJ) in lipopolysaccharide (LPS) induced in vivo and in vitro arthritis. Arthritis was induced in wistar rats by intraplantar administration of LPS (1 mg/Kg) and effect of MJ was screened in two doses (20, 40 mg/Kg, IP), indomethacin (30 mg/Kg p.o) was used as standard. The anti-nociceptive effect was evaluated through behavioral assessment viz. cold allodynia, Paw thermal hyperalgesia and Tail cold hyperalgesia on 1st, 7th, 14th, 21st and 28th day. The Myeloperoxidase (MPO), Cathepsin D (CAT-D), articular elastase (ELA), and nitrite levels were estimated in articular cartilage tissues on the 28th day. Rat paw was subjected to histopathology after radiological examination on 28th day. In vitro effect of MJ was evaluated for three concentrations (5, 10, 20 µg/ml) in LPS (1 µg/ml) stimulated CHNO001 cells. Estimation of pro-inflammatory mediators was carried using ELISA. Significant reduction in pro-inflammatory mediators was observed in MJ treated chondrocyte cells. % proteinase inhibition was assessed for 10, 50, 100, 250, 500 µg/mL and IC50 was found 266.15. MJ significantly reducesnociceptive response against hot and cold allodynia. Significant reduction in MPO, ELA, and nitrite levels was observed. The CAT-D levels significantly restored. Minimum focal mild infiltration of lymphocytes was observed at synovial area in standard and MJ treated rats. These current studies conclude that MJ has protective role in arthritis.

Hepatic perturbations provoked by azathioprine: a paradigm to rationalize the cytoprotective potential of Ficus hispida Linn.

The present study was embarked upon in an endeavor to ascertain whether Ficus hispida leaf extract (FHLE) modulates azathioprine-induced hepatic damage. Azathioprine treated rats displayed a plethora of pathological events, which include loss of hepatocellular membrane integrity, mitochondrial dysfunction, and nuclear damage; whilst FHLE pretreated rats significantly precluded these abnormalities. These data were in harmony with the transmission electron microscopic studies. Observations from this investigation directed us to propose the plausible mechanisms through which FHLE thwarts the repercussions of azathioprine-induced hepatocellular necrosis: upholding of thiol homeostasis, curtailing the membrane effects, and perpetuation of adenine nucleotide status. These data offer credence to the notion that FHLE might be a beneficial intervention in the prevention of hepatotoxicity in azathioprine therapy.

Influence of alpha lipoic acid on antioxidant status in D-galactosamine-induced hepatic injury.

D-Galactosamine (GalN)-induced liver injury is associated with reactive oxygen species and oxidative stress. In the present study, we evaluated the effect of alpha lipoic acid (ALA) supplementation on acute GalN-induced oxidative liver injury. Hepatotoxicity induced by single intraperitoneal injection of GalN (500 mg/kg body wt) was evident from increase in lipid peroxidation and serum marker enzymes (asparate transaminase, alanine transaminase, alkaline phosphatase, and lactate dehydrogenase). The decreased activities of enzymic antioxidants (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) as well as glutathione levels were the salient features observed in GalN-induced hepatotoxicity. Pretreatment with ALA (50 mg/kg body weight for 7 days) significantly precluded these changes and prevents the hepatic injury. Hence, this study clearly exemplified that ALA might be a suitable candidate against GalN-induced cellular abnormalities.

Mitigation of azathioprine-induced oxidative hepatic injury by the flavonoid quercetin in wistar rats.

ABSTRACT Azathioprine (AZA), one of the widely prescribed immunosuppressant drugs in organ transplantation and autoimmune diseases, could cause hepatotoxicity in the course of therapy. The current work was designed to assess the protective role of the dietary flavonoid, quercetin (QE), in oxidative hepatic damage induced by AZA. Adult male Wistar rats were divided into four treatment groups. Two groups were treated with single intraperitoneal injection of AZA (50 mg/kg body weight); one of these groups was pretreated with QE (50 mg/kg body weight) intraperitoneally once a day for 7 days. A vehicle treated control group and a QE control group were also included. Hepatotoxicity, evident from increased levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and gamma glutamyl transpeptidase (GGT) in serum 24 h after AZA treatment, was significantly (p < 0.05) normalized by QE pretreatment. AZA administered rats displayed declined levels of endogenous antioxidants [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione (GSH)], along with elevated levels of malondialdehyde (MDA). However, pretreatment with QE significantly precluded lipid peroxidation and maintained the activities of antioxidant defenses at a near normal status. Besides, AZA induced oxidative stress and subsequent DNA damage was effectively manifested by QE, which was confirmed by agarose gel electrophoresis. These findings highlight the salubrious effect of QE as a hepatoprotectant in AZA-induced oxidative stress mediated hepatic injury.

Immune Augmentation of Single Contact Hepatitis B Vaccine by Using PLGA Microspheres as an Adjuvant.

The present study was aimed to replace the alum type adjuvant for hepatitis B vaccine. The hepatitis B vaccine was encapsulated in poly (DL-lactide-co-glycolide) microspheres by solvent evaporation technique. The formulated microspheres were characterized in terms of morphology, particle size analysis, in vitro release study and in vivo immune response in male Wistar rats. The FT IR spectrum illustrates the characteristics bands of poly (DL-lactide-co-glycolide) microspheres and hepatitis B vaccine at 1750 cm(-1) and 1650 cm(-1), respectively. The hepatitis B vaccine loaded poly (DL-lactide-co-glycolide) microspheres were able to release antigens till day 42. Significant enhancement of specific antibodies to HBsAg was produced till day 90 after a single administration of HBsAg encapsulated poly (DL-lactide-co-glycolide) microspheres. However, the conventional alum adsorbed hepatitis B vaccine was not found to produce any significant specific antibody levels till day 90 after a single dose. The results showed that poly (DL-lactide-co-glycolide) microspheres show potential as an adjuvant for hepatitis B vaccine.