Mentha piperita silver nanoparticle-loaded hydrocolloid film for enhanced diabetic wound healing in rats.

OBJECTIVE: To investigate the role of Mentha piperita silver nanoparticle-loaded carbopol gel for enhanced wound healing in a diabetic rat model. This research further aims to explore bioactive compounds derived from Mentha piperita obtained from high altitude. METHOD: Methanolic extracts of Mentha piperita (MP), Mentha spicata (MS) and Mentha longifolia (ML) were used to synthesise silver nanoparticles (AgNP). AgNP synthesis was confirmed by ultraviolet-visible (UV-Vis) spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The antioxidant activity was assessed by 2, 2-diphenyl-1-picrylhydrazyl (DDPH) assay. Antiglycation potential was determined by measuring the fluorescent advanced glycation end products. The bioactive compound identified in the Mentha piperita methanolic (MPM) fraction through electrospray ionisation tandem mass spectrometric analysis (ESI-MS) was responsible for the highest antiglycation. The effects of MPM and MPM.AgNP-loaded Carbopol (Sanare Lab, India) on wound healing were compared in male, alloxan-induced, diabetic albino rats (200-250g), divided into control and treated groups. Effects on wound healing were assessed via histopathology. RESULTS: UV-Vis and FTIR confirmed NP synthesis with peaks for flavonoids and polyphenols. SEM and XRD explored the cubical, 30-63nm crystalline NP. The maximum antioxidant and antiglycation potential was observed in order of; MP.AgNP>MS.AgNP>ML.AgNP. The highest antioxidant activity was observed by methanolic and aqueous MP.AgNPs (88.55% and 83.63%, respectively) at 2mg.ml-1, and (75.16% and 69.73%, respectively) at 1mg.ml-1, compared to ascorbic acid (acting as a positive control, 90.01%). MPM.AgNPs demonstrated the best antiglycation potential of 75.2% and 83.3% at 1mg.ml-1 and 2mg.ml-1, respectively, comparable to positive control (rutin: 88.1%) at 14 days post-incubation. A similar trend was observed for antimicrobial activity against Bacillus subtilis, Micrococcus luteus and Escherichia coli with an inhibition zone of 21mm, 21.6mm and 24.6mm. Rosmarinic acid was the active compound present in Mentha piperita, as identified by ESI-MS. MPM.AgNP-loaded Carbopol resulted in 100% wound closure compared with control at 20 days post-wounding. In the treatment group, re-epithelialisation was achieved by day 18, compared with 25 days for the positive control group. CONCLUSION: MPM.AgNP-loaded Carbopol demonstrated safer and more effective biological properties, hence accelerating the diabetic excision wound healing process in alloxan-induced diabetic rats.

Tumor necrosis factor-alpha, prostaglandin-E2 and interleukin-1ß targeted anti-arthritic potential of fluvoxamine: drug repurposing.

Fluvoxamine, a selective serotonin re uptake inhibitor, is used to treat depression. The aim of present study was to evaluate fluvoxamine in acute (egg albumin-induced inflammation) and chronic inflammatory rat models (formaldehyde and complete Freund's adjuvant (CFA)-induced arthritis). Fluvoxamine showed highly significant (p<0.001) protective effect at dose of 50 mg/kg orally with percentage suppression 21.3% as compared to disease control group in acute model. Likewise, formaldehyde-induced arthritic experiment confirmed the significant (p<0.001) anti-arthritic behavior, showed by fluvoxamine (50 mg/kg orally) throughout the study. Moreover, In CFA-induced model, the higher dose (fluvoxamine 50 mg/kg) exhibited highly significant (p<0.001) decrease in paw thickness and arthritic score with significant increase in weight of animals from 123.8± 1.934 g to 130.2± 1.655 g, significantly decreased the level of RF and CRP to level of 12.0±0.707 and 11.40±0.50 respectively and restoration of SOD, CAT (69.8±1.5, 72.0±1.4 respectively). Furthermore, the level of TNF-α, PGE2, and IL-1ß (147.0±2.0, 406.8±2.5, and 93.8±1.3 respectively) in arthritic animals was reduced to significant (p<0.001) level (53.8±1.3, 205±3.6, and 42.0±1.4 respectively) after treatment with fluvoxamine. Furthermore, molecular docking of fluvoxamine against TNF-α, PGE2, and IL-1ß protein targets showed good binding energies which hereby from computational studies proves our compound anti-inflammatory potential. In addition, absorption, distribution, metabolism, excretion, and toxicity (ADMET) studies reveled that fluvoxamine has very good pharmacokinetic profile with no specific hepatic toxicity and good absorption level. In addition, the skin sensitization test in vitro human cell line activation test (h-CLAT) and KeratinoSens have revealed that isolated flavone is not skin sensitive with confidence score of 59.6% and 91.6%. The current findings validated the anti-arthritic potential of fluvoxamine but it should be recommended for clinical investigation in future research.

Geranium wallichianum D. Don Ex Sweet Ameliorates Rheumatoid Arthritis by Curtailing the Expression of COX-II and Inflammatory Cytokines as Well as by Alleviating the Oxidative Stress.

Geranium wallichianum D. Don ex sweet traditionally been used as home remedy for backaches, joint pain, colic, and rheumatism. The objective of this study was to investigate the therapeutic benefits of plant in an adjuvant-induced arthritis paradigm. Immune-mediated rheumatoid arthritis was developed by injecting complete Freund's adjuvant (CFA) into the hind paws of rats and the aqueous methanolic crude extract was administered. The animals were physically monitored for changes in paw edema size and arthritic score. Hematological parameters and systemic inflammatory indicators evaluated. Genetic expressions of tumor necrosis factor (TNF-α), interleukins (IL-1ß, IL-6), necrosis factor (NF-κB), and cyclooxygenase (COX-II) enzyme were studied using real-time qPCR. PGE2 levels in blood were quantified through Enzyme Linked Immunosorbent Assay (ELISA). On the 14th day, Immunoglobulin E (IGE) exhibited a substantial decline in paw edema and arthritic score. At the doses of 500 mg/Kg (P ≤ .05) and 1000 mg/Kg (P ≤ .001), IGE significantly reduced TNF-α, interleukins, and COX-II mRNA expression. IGE significantly lowered the MDA levels at the doses of 500 and 1000 mg/Kg (13.18 ± .70 and 9.04 ± .26 µM/L respectively) as compared to arthritic control (30.82 ± 1.12 µM/L) group. IGE significantly improved the antioxidant enzyme activities of CAT and SOD (P ≤ .001) in treated animals. TNF-α, interleukins, and COX-II mRNA expression were also significantly reduced at the doses of 300 (P ≤ .05), 500 (P ≤ .01) and 1000 mg/Kg (P ≤ .001) which were expressed as fold changes. This study shows that Geranium wallichianum D. Don ex sweet has a strong potential to alleviate immune-mediated arthritis by lowering oxidative stress and downregulating the proinflammatory cytokines signaling mechanisms.

Caralluma tuberculata exhibits analgesic and anti-arthritic potential by downregulating pro-inflammatory cytokines and attenuating oxidative stress.

Caralluma tuberculata N.E. Brown (Common name: Chongan), belonging to the family Asclepiadaceae is distributed widely in hilly areas of Dir, Swat, Kohat and in plain lands of Punjab, Pakistan. The plant has been used as a source of vegetable as well as home remedy for headache, muscle spasms and rheumatism. The present study was proposed to investigate the analgesic, anti-inflammatory and anti-arthritic potential of the aqueous methanolic extract of C. tuberculata (ICE). The dried shoots of plant were used to prepare aqueous methanolic extract (30:70) by 3 days thrice maceration and filtration followed by evaporation under reduced pressure. ICE was screened for the presence of phytochemicals using preliminary phytochemical analysis and HPLC. The antioxidant potential was evaluated through DPPH assay. Analgesic potential of ICE was studied using hot plate and tail immersion methods, and anti-inflammatory activity was performed using turpentine oil and carrageenan-induced inflammation models, in wistar albino rats. Formaldehyde-induced and Complete Freund's Adjuvant-induced arthritis models were used for the assessment of anti-arthritic activity of ICE and its effects on serum levels of PGE-2 as well as gene expression levels of pro-inflammatory cytokines were studied. ICE displayed a dose-dependent (300-1000 mg/Kg p.o.) analgesic effect in hot plate (maximum retention time of 10.87 and 13 s) and tail immersion (response time of 11 and 13.64 s) tests at the doses of 500 and 1000 mg/Kg, respectively. The extract exhibited a significant decrease in paw inflammation of rats at the doses of 500 and 1000 mg/Kg as compared to the disease control group. ICE also exhibited a remarkable decline in arthritic score and a dose-dependent drop in serum levels of prostaglandin E2. There was a significant suppression in the expression of TNF-α, IL-1ß, IL-6, NF-κB and cyclooxygenase enzyme in treatment groups. This study concludes that Caralluma tuberculata exhibits strong analgesic, anti-inflammatory, antioxidant and anti-arthritic activities thus upholding the vernacular use of the plant for pain and rheumatism.

DNA binding and cleavage, BRCA1 gene interaction, antiglycation and anticancer studies of transition metal complexes of sulfonamides.

A series of 4-((4-methylphenylsulfonamido)methyl)cyclohexanecarboxylic acid (NaMSCCA) transition metal complexes [Cu(II), Zn(II), Ni(II), Mn(II), and Co(II)] have been synthesized by precipitation method. The characterization was done by physical techniques, FT-IR spectroscopy, mass spectrometry, and NMR spectroscopy. The molecular structures of nickel (II) AZ-3 and cobalt (II) AZ-5 complexes were determined by the X-ray diffraction technique and found to crystallize in the triclinic space group P-1. The coordination geometry around the central nickel (AZ-3) and cobalt (AZ-5) atoms was square planar bipyramidal. Molecular docking was performed with duplex DNA of sequence d(CGCGAATTCGCG)2 DNA to determine the probable binding mode of compounds. Then these synthesized compounds were used to perform DNA cleavage activity through the agarose gel electrophoresis method. Among the compounds, compounds AZ-1 and AZ-2 exhibited good nuclease activity. The DNA sequence of breast-cancer suppressor gene 1 (BRCA1) was amplified through PCR and interaction studies of compounds AZ-1 and AZ-2 were performed through gel electrophoresis and fluorescence emission spectroscopy. The expression analysis of the BRCA1 gene was also performed to quantify the expression relative fold change (2^-(∆∆CT)) after treatment with compounds. All synthesized compounds were evaluated for their antioxidant and antiglycation activities and AZ-2 exhibited excellent results. The molecular docking study of these compounds was performed against the protein structure of advanced glycation end products to support the experimental results. Anticancer activity of compounds was performed through MTT assay. Copper and zinc complexes depicted the highest anticancer activity against human breast adenocarcinoma (MCF7) and human corneal epithelial cell (HCEC) cell lines.

Pharmacological support to anti-arthritic prospective of physostigmine: a new approach.

Rheumatoid arthritis (RA) is a slowly progressing inflammatory autoimmune disease. Several features are involved in the RA pathogenesis in addition to environmental and genetic factors. Previously it has been reported that acetyl cholinesterase (AChE) activity is enhanced in old age and may contribute in the progression of RA. The current experimental work was projected to assess the activity of physostigmine (a cholinesterase inhibitor) for treatment of RA. In vitro and in vivo approaches were used for such evaluation. However, enzyme linked immunosorbent assays (ELISA) was performed to determine the concentrations of Prostaglandins E2 (PGE2) and tumor necrosis factor-α in arthritic rats after treatment with physostigmine. Moreover, anti-oxidant assays were employed to calculate the level of super oxide dismutase (SOD) and catalase peroxidase (CAT) in tissue of treated animals. The results claimed the dose dependent protective and stabilizing effect of physostigmine on denaturation of albumin (egg and bovine serum) protein and human red blood cell membrane, respectively, through in vitro studies. Furthermore, the physostigmine (10 and 20 mg/kg) significantly (p < 0.001) reduced the swelling of paw after induction of arthritis with formaldehyde or complete Freund's adjuvant (CFA) as compared to arthritic control animals. Moreover, significant (p < 0.001) reduction in the levels of inflammatory markers (PGE2 and TNF-α) at doses of 10 and 20 mg/kg of physostigmine has been observed in ELISA test. Likewise, there was a prominent rise in levels of SOD and CAT in animals treated with physostigmine. These findings pharmacologically conclude the anti-arthritic effect of physostigmine.

Antioxidant and antiglycation activities of traditional plants and identification of bioactive compounds from extracts of Hordeum vulgare by LC-MS and GC-MS.

Glycation has been involved in Schiff base reaction lead to hyperglycemia at cellular level. The current study aimed to identify the bioactive compounds from selected folkloric plants for their antiglycation and antioxidant potential. Methanol extracts demonstrated the highest activities, therefore, it was further fractionated using n-hexane, dichloromethane, ethyl acetate, and methanol solvents to isolate the nonpolar compounds from the Hordeum vulgare. Moreover, n-hexane and dichloromethane fractions of H. vulgare demonstrated the best antioxidant (61.58% and 62.89%) and antiglycation activities (72.52% and 61.52%) at 2 mg/ml, respectively. Analytical techniques of LC-MS and GC-MS were employed for identification of bioactive compounds; Biochanin A in dichloromethane (DCM) and Vitamin E in n-hexane fractions. There was a strong correlation between antioxidant and antiglycation activities (r = 0.97 and r = 0.96) of DCM & n-hexane fractions of H. vulgare. Findings of this study established the role of Biochanin A and Vit E from H. vulgare as potent antiglycation agents. PRACTICAL APPLICATIONS: The results of this study confirmed the potential role of Black Barley has involved in the inhibition of protein glycation, which can be the potential treatment to reduce the complications of Diabetic Patients. The Black Barley has a rich source of identified compounds Biochanin A and Vitamin E. We can use this plant as a staple food in curing the severity of diabetes. The other practical approach is to use this plant as an ingredient of different food products. The extraction of identified bioactive compounds from the plant will be a good and cheap source of the treatment.