Anti-inflammatory Assay Research Articles

Egyptian Citrus Essential Oils Recovered from Lemon, Orange, and Mandarin Peels: Phytochemical and Biological Value

Citrus peels are an important by-product of citrus processing industries, but a large part is considered waste. There has been increased attention in the last five years on these industrial by-products, especially those containing residual essential oils (EOs). Lemon, orange, and mandarin peels from Egypt were subjected to hydro-distillation to obtain EOs, which were analyzed via mass spectrometry (GC/MS) and by building Global Natural Products Social Molecular Networking (GNPS-MN) for the purpose of visually exploring the volatile components of citrus species. The constructed MN revealed that D-Limonene, α-pinene, and β-pinene are the dominant volatile constituents in the three Egyptian citrus species. The EOs from three citrus peels exhibited promising activities as antioxidants using two tested methods: 1,1-diphenyl-2-picryl-hydrazil (DPPH) and nitric oxide (NO) compared with vitamin C. Lemon EO proved excellent antimicrobial activity against Gram-positive and negative bacteria. Additionally, the three citrus EOs showed good activities against the yeast Candida albicans. Regarding the anti-inflammatory assay, the three citrus EOs showed promising activities as COX-1 and COX-2 inhibitors. This study concludes that EOs extracted from citrus peel waste can be valorized as an innovative strategy for food preservation or may be incorporated in cosmetics and pharmaceutical formulations in alignment with circular economy principles.

Evaluation of Antioxidant, Anti-inflammatory, and Antimicrobial Activities of Raspberry Fruit Extract: An In Vitro Study.

Introduction Rubus idaeus (R. idaeus)is an edible fruit thatcontains numerous significant bioactive compounds that hold important biological properties and are categorized as nutraceuticals owing to the health benefits it imparts including decreasing the risk of chronic diseases like diabetes mellitus, cancer, heart disease, and many other diseases.The objective of the present research was to explorethe antimicrobial, antioxidant, and anti-inflammatory characteristics of the aqueous raspberry extractthrough in vitro assays. Materials and methods R. idaeus aqueous extract was prepared and examined for its antimicrobial activity against Streptococcus mutans (S. mutans) bacteria and Candida albicans (C. albicans) fungi using the agar-well diffusion method, and the antioxidant activity was evaluated using the DPPH (2, 2-Diphenyl-1-picrylhydrazyl hydrate) radical scavenging assay and the hydrogen peroxide radical scavenging assay. The anti-inflammatory activity of the prepared extract was investigated using bovine serum albumin (BSA) and egg albumin denaturation assays. Results R. idaeus fruit extract displayed strong antimicrobial activity at a higher concentration of 100 µL with a 26 mm zone of inhibition against Streptococcus mutans and 24 mm for Candida albicans. The extract showed 87.42% hydrogen peroxide free radical scavenging activity and inhibited 91.12% DPPH free radicals at the highest concentration of 50 µg/mL. The extract showed effective anti-inflammatory activity by preventing the denaturation of bovine serum albumin (80%) and egg albumin proteins (77%) at the highest concentration of 50 μg/mL.The free radical scavenging activity positively correlates with the increased concentration of the prepared extract against DPPH and hydrogen peroxide free radicals, thus showing the raspberry extract's potent antioxidant activity. Similarly, the anti-inflammatory assay result shows that the prepared raspberry aqueous extract has excellent anti-inflammatory activity by preventing the denaturation of bovine serum albumin and egg albumin protein in a dose-dependent manner. Conclusion The meticulously prepared raspberry extract exhibited noteworthy antimicrobial, antioxidant, and anti-inflammatory characteristics, and owing to its astounding therapeutic benefits it holds a tremendous promise as a natural alternative in the field of oral medicine especially in the management of oral mucosal lesions, oral potentially malignant lesions such as lichen planus and leukoplakia, candidiasis, oral cancer and oral mucositis. Further animal studies and clinical trials are recommended to fully reap the therapeutics potential of raspberry extract.

Protective and Ameliorative Effects of Hydroethanolic Extract of Piper nigrum (L.) Stem against Antiretroviral Therapy-Induced Hepatotoxicity and Dyslipidemia in Wistar Rats.

Antiretroviral therapy (ART) has revolutionized the lives of people living with HIV/AIDS by overall improving their quality of life and increasing life expectancy. However, ART-associated hepatotoxicity and metabolic disorders in HIV/AIDS patients are growing concerns to clinicians, especially due to the long-term use of the drugs. This study reported on the phytochemical and pharmacological profile of hydroethanolic extracts of Piper nigrum stem (PNS) and evaluated its protective effect against tenofovir/lamivudine/efavirenz (TLE)-induced hepatotoxicity and dyslipidemia in Wistar rats. Cytotoxic, antioxidant, and anti-inflammatory assays were performed on PNS. Thirty-six rats divided into 6 groups of 6 animals/group were administered: distilled water, 17 mg/kg TLE, 17 mg/kg TLE and 100 mg/kg silymarin, 17 mg/kg TLE, and Piper extract (200 mg/kg, 400 mg/kg, or 800 mg/kg) orally for 28 days. The body weight of animals was recorded every 7 days. On Day 29, the rats were sacrificed, and blood samples were collected for hematological and biochemical tests. Portions of the liver and kidneys were collected for histological evaluation, while liver homogenates were prepared from the rest to measure antioxidant enzymes. PNS possessed in vitro cytotoxic, antioxidant, and anti-inflammatory activities. A significant decrease (p < 0.05) in the body weight of rats treated with PNS was observed. A significant high platelet count (p < 0.05) was observed in the PNS800 mg/kg group. A considerable decrease in alkaline phosphatase and triglycerides was observed in the silymarin and PNS group compared to the TLE-only group. The findings also show a significant increase in catalase and glutathione in the TLE-only group compared to the normal group, while SOD decreased. Histological observations revealed normal hepatic and renal tissues in the silymarin, and PNS-treated groups compared to the normal control, while leucocyte infiltration was observed in the TLE-only group. These results suggest that PNS extract possessed antioxidant activity that alleviated TLE-induced toxicity. Further studies are necessary to understand the pharmacokinetic interactions between ART and PNS.

In-Vitro Evaluation of Cytotoxicity and Antiinflammatory Potential of Hypnea Musciform (Rhodophyta) Collected from the Coastal Area of Karachi, Pakistan

Background: The extensive littoral zones of Karachi, Pakistan, are home to a diverse marine phytoplankton population, including the underutilized Hypnea musciformis. This red algae is an unexplored potential source of bioactive compounds for pharmacological use. Objective: This study aimed to evaluate the cytotoxicity and anti-inflammatory potential of the aqueous extract of Hypnea musciformis collected from the coastal areas of Karachi. Methods: Hypnea musciformis was collected from November 2019 to February 2021, with environmental parameters such as seawater temperature, pH, and salinity recorded. The collected samples were dried, powdered, and extracted in water. The cytotoxicity was assessed using a brine shrimp lethality test across a range of concentrations (1, 10, 100, and 1000 µg/ml), determining the LC50 value. Anti-inflammatory activity was evaluated via a membrane stabilization method on human red blood cells (HRBC), comparing the effects with the standard drug Diclofenac at concentrations ranging from 100 to 200 µg/ml. Results: The brine shrimp lethality assay showed a dose-dependent increase in mortality with the highest concentration exhibiting 83% mortality, yielding an LC50 value of 550 µg/ml. The anti-inflammatory assay demonstrated that the aqueous extract of Hypnea musciformis provided a concentration-dependent protection with a maximum protection percentage of 78.64% at 200 µg/ml, while the control substance, Diclofenac, displayed higher efficacy. Conclusion: The aqueous extract of Hypnea musciformis possesses significant cytotoxic and anti-inflammatory activities, suggesting its potential as a source of natural compounds for therapeutic purposes. However, further studies are required to elucidate the mechanisms of action and to confirm the safety and effectiveness of the extract in clinical applications.

New sesquiterpenoids from Biscogniauxia sp.

Five new sesquiterpenoids, including a campherenane-type (1), a bergamotane-type (2), a drimane-type (3), and two bisabolane-type (5-6) sesquiterpenoids have been isolated from Biscogniauxia sp. 71-10-1-1. Their structures were determined by spectroscopic analyses, quantum chemical ECD calculations,13C chemical shifts calculations, and X-ray crystallography. This is the first report of campherenane-type and drimane-type sesquiterpenoids from Biscogniauxia. Furthermore, the anti-inflammatory assays of all compounds are evaluated, and the results showed that compounds 3 and 7 exhibited the effects against the production of the pro-inflammatory cytokine TNF-α.

Anti-rheumatic potential vis-à-vis aconitine and hypaconitine content analysis in different Aconitum spp. from Sikkim Himalayas (India).

Aconitum spp. are important medicinal plants mentioned in Ayurveda as Ativisa or Vatsanabha. The present study aims to evaluate anti-rheumatic potential in seven Aconitum species and correlation with aconitine and hypaconitine content. Anti-rheumatic potential was analyzed through in vitro xanthine oxidase inhibition, anti-inflammatory and ROS scavenging assays; and quantification of aconitine and hypaconitine with RP-HPLC method validated as per ICH guidelines. The findings reveal that A. palmatum possessed the most promising response (IC50 =12.68±0.15 μg/ml) followed by A. ferox (IC50 =12.912±1.87 μg/ml) for xanthin oxidase inhibition. We observed a wide variation in aconitine and hypaconitine content ranging from 0.018 %-1.37 % and 0.0051 %-0.077 % respectively on dry weight basis. Aconitine and hypaconitine showed moderate positive correlation (r=0.68 and 0.59 respectively) with anti-rheumatic potential. The study identifies potential alternative species of Aconitum that can help in sustainable availability of quality raw material.

Evaluation of antimicrobial, antioxidant, and anti-inflammatory abilities of sustainably synthesized Co3O4 NPs

The cobalt oxide nanoparticles (Co3O4 NPs) sustainably synthesized by Citrus tangerina leaf aqueous extract and it was characterized by standard analytical experiments including UV–vis spectra, FT-IR, DLS, and SEM analyses. Furthermore, their antibacterial, antifungal, antioxidant, and anti-inflammatory competence were investigated through in-vitro approach. The aqueous extract derived from C. tangerina leaf has the potential to synthesize the Co3O4 NPs and it was evidenced by peak appeared at 399 nm under UV–vis spectrum study. Furthermore, FTIR analysis results demonstrated that the occurrence of most active functional groups (alcohols (C–O), phenols (C–O), carbonyl (C=O), and amide (CO = )). The DLS and SEM analyses revealed that the average size as well as shape of C. tangerina leaf extract mediated Co3O4 NPs were found as ranging between 90 and 130 nm and octahedral in shape respectively. The C. tangerina leaf fabricated Co3O4 NPs demonstrated dose dependent activity for antibacterial (e.g. S. aureus 6538 > E. coli O157 > K. pneumoniae 342 > B. subtilis 6633 at 200 μg mL−1 concentration), antifungal, antioxidant, and anti-inflammatory assays. The obtained findings conclude that the C. tangerina leaf aqueous extract has the potential to synthesize the Co3O4 NPs and synthesized particle has considerable selective biomedical applications.

Fabrication and physicochemical investigation of pH-responsive alginate/pectin hybrid network hydrogel for improved stability and controlled release of diallyl thiosulfinate

The present investigation explores the formulation and fabrication of alginate-pectin (with varying degree of methoxylation) hybrid network hydrogel beads through ionotropic gelation process as a biocompatible carrier material for encapsulating diallyl thiosulfinate (allicin). The primary aspiration of this research is to overcome allicin's inherent instability which restricts its use in functional food fortification and medicinal products as a bioactive compound with several intriguing health-promoting characteristics. Fabrication of the beads were done through structured parameter optimization. Chemical interaction between allicin and biopolymer matrix was assessed by FTIR-ATR. The ARGE entrapped hydrogel beads had a near spherical shape with sphericity factor ∼1.0 with an average diameter of ∼3.5 mm in the hydrated state (density ∼1.06 g/cm3). The beads showed encapsulation efficiency of ∼70.8 %. The beads efficaciously reduced degradation (<5 %) and stabilized ampule amount of allicin for a prolonged time. Various physicochemical characterizations of the produced beads were done. Further, in vitro release kinetics of allicin from beads were observed in simulated gastrointestinal environment and it showed a sustained release pattern. The release behavior of allicin exhibited a higher release at intestinal pH (∼88 %) than stomach pH (∼35 %). The formulation expressed commendable antimicrobial activity against gram positive and negative bacteria in both solid and liquid media. Moreover, the encapsulation process retained significant antioxidant potential (>60 %). Anti-inflammatory and hemolysis assay expressed admirable biocompatibility. Hence, the results suggest that the fabricated formulation is suitable for being a drug carrier material for allicin while accentuating its aptitude for upholding hemocompatibility, allicin’s concentration, sustained release and bioactivity.

Optimization of silver nanoparticles synthesis via Plackett-Burman experimental design: in vitro assessment of their efficacy against oxidative stress-induced disorders.

Nanoparticles possess remarkable biological activities owing to their small size and large surface-to-volume ratio. Given the increasing adoption of environmentally sustainable practices in silver nanoparticle (AgNP) fabrication, this study presents a simple lab-scale green synthesis of AgNPs using banana peels. Large amounts of banana peels are disposed off in Pakistan every day. As the fruit is available throughout the year and contains many active components with potent biological activities, we aimed to synthesize silver nanoparticles using its peel, through an energy-efficient and inexpensive route. The synthesis was optimized according to the Plackett-Burman design (PDB) of experiments, which helped identify significant factors and saved time and resources. For characterization, UV-Vis spectroscopy and SEM-EDX analysis were performed, revealing spherical particles in the 45-65 nm size range. To investigate functional groups, FT-IR analysis was performed, revealing the presence of N-C[double bond, length as m-dash]O amide I bonds of proteins, C-H bonds of tannins and C-O bonds involved in the capping and stabilization of nanoparticles. The free radical scavenging property of banana peel-mediated silver nanoparticles (BP-AgNPs) was studied against 2,2-diphenyl-1-picrylhydrazyl (DPPH), and the antioxidant potential was found to be 79% at 500 μg mL-1 concentration. The efficacy of BP-AgNPs with respect to certain biological activities were studied through anti-inflammatory assays, which demonstrated better results compared to a standard drug, and an anti-glycation assay, wherein only 4% of AGEs were formed, demonstrating 96% of AGE inhibition in vitro. The findings not only demonstrated the effectiveness of the PBD approach but also highlighted the potent property of BP-AgNPs against disorders associated with oxidative stress.

Neolignan Glycoside and Other Constituents From the Leaves of Ligustrum sinense and Their Anti-Inflammatory Activity

Objectives: The aim of the project was the isolation, structure elucidation, and anti-inflammatory evaluation of compounds from the leaves of Ligustrum sinense Lour. Methods: Chromatographic techniques were used to isolate anti-inflammatory compounds from the methanol extract of L sinense leaves. The structures of compounds were elucidated by analyses of mass spectra, nuclear magnetic resonance data, and Circular dichroism spectra. Compounds were evaluated using anti-inflammatory assays. Results: One new neolignane glucoside, neoligustrume A (1), along with eleven known compounds, ligujaponoside A (2), ligujaponoside B (3), ligustroside (4), oleuropein (5), ligustaloside A dimethyl acetal (6), ligustaloside B dimethyl acetal (7), olivil (8), (+)-cycloolivil (9), kaempferol 3- O-β-D-glucopyranoside-7- O-α-L-rhamnopyranoside (10), oleanoic acid (11), and hydroxytyrosol (12) were isolated from the methanol extract of L sinense leaves. Compounds 2, 3, and 12 showed significant inhibitory NO production with IC50 values of 16.3 ± 0.5, 18.2 ± 1.1, and 15.7 ± 1.9 μM, respectively. Compounds 4-7 and 11 showed moderate inhibitory NO production with IC50 values ranging from 21.4 to 45.0 µM. Moreover, compound 12 showed the most TNF-α inhibition with 58.1 ± 5.7% at a concentration of 50 μM. Conclusions: One new and 11 known compounds were isolated from L sinense. This is the first report of compounds 2-9 from L sinense. Compounds 2, 3, and 12 showed significant inhibitory NO production with IC50 values of 16.3 ± 0.5, 18.2 ± 1.1, and 15.7 ± 1.9 μM, respectively. Compound 12 could be the anti-inflammatory source.

Exploring the Anti-Inflammatory Potential of Phytochemicals from Anvillea radiata: In Vitro Assay, Molecular Docking, and Molecular Dynamics Simulations

Abstract This study delves into the comprehensive evaluation of phytochemicals extracted from Anvillea radiata, with a focus on their potential anti-inflammatory applications. High-performance liquid Chromatography analysis reveals the dominance of catechin in the aqueous extract, alongside other compounds such as syringic acid, 3,4-dihydroxybenzoic acid, and vanillic acid, as well as quercetin-3-β-D-glucoside and rutin in smaller quantities. An in vitro anti-inflammatory assay demonstrates the superior effectiveness of A. radiata aqueous extract at lower concentrations compared to Aspirin, raising possibilities for reduced side effects and enhanced cost-effectiveness. Drug-likeness predictions emphasize the potential of catechin, syringic acid, and vanillic acid for drug development due to their molecular characteristics. Toxicity risk assessment indicates that catechin, quercetin-3-β-D-glucoside, and rutin exhibit low toxicity risks and favorable drug-likeness properties. Molecular docking analysis highlights promising interactions between the identified compounds and crucial anti-inflammatory (COX-2, MPO, IL1β, IL6, and TNFα.) target proteins. Notably, catechin demonstrates a strong binding affinity to these proteins. Molecular dynamics simulations further support catechin’s potential, revealing significant deformability in complexes with COX-2 and MPO, indicating high structural flexibility. The variance map shows higher cumulative variances and the elastic network map yields satisfactory outcomes, further reinforcing the potential of A. radiata aqueous extract compounds, with catechin as a standout candidate for anti-inflammatory drug development.

Size Matters: Influence of Particle Size on Antioxidant, β-Glucan, and Anti-Inflammatory Potential in Pleurotus floridanus (Agaricomycetes).

Cellular damage resulting from elevated levels of free radicals can lead to persistent health issues. Pleurotus floridanus, an edible white oyster mushroom, is rich in β-glucans with potent antioxidant and anti-inflammatory properties. In this research, we examined the β-glucan content, total phenolic content, as well as antioxidant and anti-inflammatory potential of hot water extracts with varying particle sizes (< 75, 75-154, 154-300, and 300-600 μm) of both whole and sliced fruiting bodies of P. floridanus. The findings revealed that the в-glucan content increased as the particle size increased, although no significant differences were observed. Conversely, smaller particle sizes (< 75 μm) of whole and sliced fruiting bodies of P. floridanus exhibited higher phenolic content, 2,2-diphenyl-1-picryl-hy-drazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, and reducing ability compared with larger particle size (> 75 μm). Of the four samples (AW2, AW3, AS1, and AS2) with the highest antioxidant activity selected for anti-inflammatory assays, all demonstrated the ability to reduce nitric oxide and tumor necrosis factor-alpha levels, but did not enhance interleukin-10 expression in lipopolysaccharide-stimulated RAW264.7 cells. Interestingly, particle size < 75 to 300 μm did not appear to influence the anti-inflammatory activity, because no significant differences were observed among the particle sizes. Therefore, a particle size < 300 μm in a P. floridanus hot water extract could serve as a valuable source of antioxidant and anti-inflammatory compounds to counteract the harmful effects of free radicals.

Comparative Anti-Inflammatory Activity of Rutin-Based Mouthwash and Diclofenac Sodium: An in-Vitro Evaluation

Background: One of the most common oral inflammatory conditions is Gingivitis. It involves the inflammation of gingiva supporting the tooth. Onset of gingivitis is due to the recurrent deposition of plaque and its inefficient removal. Use the right oral dentifrice and mouthwash to keep the inflammation at bay. Rutin is a natural substance extracted from citrus fruits. This citrus flavonoid is known to possess antimicrobial, antioxidant, and cytotoxic properties. Aim: This study aims to formulate a mouthwash with rutin and compare its anti-inflammatory potential with an already commercially anti-inflammatory agent. Materials and Methods: For this in-vitro study two different anti-inflammatory assays were performed using different substrates. The first test performed was the bovine serum albumin anti-inflammatory test and the second one was the egg albumin denaturation test. Results: The results suggest that the rutin mouthwash made in our study shows a similar anti-inflammatory activity as the Diclofenac sodium based formulation. Rutin mouthwash and the Diclofenac sodium have exhibited a non-significance in the percent protein anti-denaturation with the increase in the concentrations ranging from to . Conclusion: Rutin-based mouthwash possesses satisfactory anti-inflammatory activity compared to standard commercial mouthwash.

Anti-inflammatory activity of verbascoside- and isoverbascoside-rich Lamiales medicinal plants

Verbascoside and isoverbascoside are two active phenylethanoid glycosides mainly found in plants of the order Lamiales. This study analyzes the verbascoside and isoverbascoside levels and the total phenolic contents in the water and ethanolic extracts of 20 medicinal plants of the order Lamiales commonly used in Thailand. The related bioactivities, including the antioxidant activity via the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reduction activity potential assays and anti-tyrosinase and -inflammatory activities via the cyclooxygenase and nitric oxide assays are also investigated. The extracts of several plant species, including Barleria prionitis, B. lupulina, Rhinacanthus nasutus, Orthosiphon aristatus, and Nicoteba betonica, exhibit high verbascoside and isoverbascoside content levels. The correlation analysis between the bioactive activities and the active compounds demonstrates a significant association between the verbascoside level in the water extracts and both the DPPH antioxidant activity and the nitric oxide level in the anti-inflammatory assays. This study provides the first report on the verbascoside and isoverbascoside quantification of several plant samples. The findings provide valuable insights for future research on lesser-studied plants possessing high verbascoside and isoverbascoside levels, which exhibit promising anti-inflammatory activities.

Unveiling the therapeutic potential: Evaluation of anti-inflammatory and antineoplastic activity of Magnolia champaca Linn’s stem bark isolate through molecular docking insights

Magnolia champaca Linn. has traditionally been used for medicinal activity in Asia for treating various chronic diseases as well as a source of food, medicines, and other commodities. Due to the long-used history of this plant, the present study was designed to explore the in vitro, in vivo and in silico anti-inflammatory and antineoplastic properties of the methanolic extract and fractions and the pure compound isolated from the most active chloroform fraction (CHF) of the stem bark of the plant. The isolated compound from the most active CHF was characterized and identified as a glycoside, trans-syringin, through chromatographic and spectroscopic (1H-NMR and 13C-NMR) analyses. In the in vitro anti-inflammatory assay, CHF was most effective in inhibiting inflammation and hemolysis of RBCs by 73.91 ± 1.70% and 75.92 ± 0.14%, respectively, induced by heat and hypotonicity compared to standard acetylsalicylic acid. In the egg albumin denaturation assay, CME and CHF showed the highest inhibition by 56.25 ± 0.82% and 65.82 ± 3.52%, respectively, contrasted with acetylsalicylic acid by 80.14 ± 2.44%. In an in vivo anti-inflammatory assay, statistically significant (p < 0.05) decreases in the parameters of inflammation, such as paw edema, leukocyte migration and vascular permeability, were recorded in a dose-dependent manner in the treated groups. In the antineoplastic assay, 45.26 ± 2.24% and 68.31 ± 3.26% inhibition of tumor cell growth for pure compound were observed compared to 73.26 ± 3.41% for standard vincristine. Apoptotic morphologic alterations, such as membrane and nuclear condensation and fragmentation, were also found in EAC cells after treatment with the isolated bioactive pure compound. Such treatment also reversed the increased WBC count and decreased RBC count to normal values compared to the untreated EAC cell-bearing mice and the standard vincristine-treated mice. Subsequently, in silico molecular docking studies substantiated the current findings, and the isolated pure compound and standard vincristine exhibited −6.4 kcal/mol and −7.3 kcal/mol binding affinities with topoisomerase-II. Additionally, isolated pure compound and standard diclofenac showed −8.2 kcal/mol and −7.6 kcal/mol binding affinities with the COX-2 enzyme, respectively. The analysis of this research suggests that the isolated bioactive pure compound possesses moderate to potent anti-inflammatory and antineoplastic activity and justifies the traditional uses of the stem bark of M. champaca. However, further investigations are necessary to analyze its bioactivity, proper mechanism of action and clinical trials for the revelation of new drug formulations.

Evaluation of Opuntia-carrageenan superporous hydrogel (OPM-CRG SPH) as an effective biomaterial for drug release and tissue scaffold

The process of wound healing involves complex interplay of systems biology, dependent on coordination of various cell types, both intra and extracellular mechanisms, proteins, and signaling pathways. To enhance these interactions, drugs must be administered precisely and continuously, effectively regulating the intricate mechanisms involved in the body's response to injury. Controlled drug delivery systems (DDS) play a pivotal role in achieving this objective. A proficient DDS shields the wound from mechanical, oxidative, and enzymatic stress, against bacterial contamination ensuring an adequate oxygen supply while optimizing the localized and sustained delivery of drugs to target tissue. A pH-sensitive SPH was designed by blending two natural polysaccharides, Opuntia mucilage and carrageenan, using microwave irradiation and optimized according to swelling index at pH 1.2, 7.0, and 8.0 and % porosity. Optimized grade was analyzed for surface hydrophilicity-hydrophobicity using OCA. Analytical characterizations were performed using FTIR, TGA, XRD, DSC, reflecting semicrystalline behavior. Mechanical property confirmed adequate strength. In vitro drug release study with ciprofloxacin-HCL as model drug showed 97.8 % release within 10 h, fitting to the Korsmeyer-Peppas model following diffusion and erosion mechanism. In vitro antimicrobial, anti-inflammatory assays, zebrafish toxicity, and animal studies in mice with SPH concluded it as a novel biomaterial.

Heterobivalent Peptide Agonists Comprising Covalently Linked Protease Activated Receptor 1 and Protease Activated Receptor 3 Peptides Mimic the Cytoprotective Cell Signaling Activities of Activated Protein C

Introduction: Activated protein C (APC) exhibits both anticoagulant and cytoprotective cell signaling actions, including anti-inflammatory and anti-apoptotic activities and endothelial barrier stabilization. Extensive in vitro and in vivo data for APC's cell signaling benefits identified requirements for two G-protein coupled receptors (GPCR), protease-activated receptor (PAR) PAR1 and PAR3 which are cleaved by APC at the noncanonical sites (R46 and R41, respectively). Agonism of PARs following proteolysis of the N-terminal tail is driven by the newly revealed N-terminus and related synthetic peptides, e.g., for APC, PAR1 peptides (P1) beginning with N47 or PAR3 peptides (P3) beginning with G42. We found that the non-canonical P1-47-66 and P3-42-65 peptides mimic APC in that each is anti-inflammatory by reducing NLRP3-driven caspase-1 activity in human THP-1 cells (Healy LD et al, J Thromb Haemost 2021;19:269-280). Remarkably, these two peptides act synergistically to attenuate activity of caspase-1 at low concentrations where either peptide alone has little activity. Thus, we undertook studies of P1 peptides that were covalently linked to P3 peptides for ability to mimic APC's anti-inflammatory and endothelial barrier stabilization actions. Methods: Monocytes (THP-1 cells) were differentiated into macrophages using PMA, then pretreated with APC or peptides prior to stimulation with lipopolysaccharide and adenosine triphosphate to induce caspase-1 whose activity was measured using a luminescent caspase-1 substrate. Endothelial barrier stability was monitored for transendothelial electrical resistance (TEER) using an electric cell-substrate sensing system (iCelligence apparatus) when cultured human EAhy926 or murine aortic endothelial cells (EC) were exposed to thrombin treatment (0.25 nM) plus or minus treatment with P1 and/or P3 peptides or G10-related peptides. Synthetic P1 and P3 sequence-derived peptides of various compositions were made, and bivalent P1:P3 peptides were made using oligo-Gly-peptide linkers containing from 4 to 14 Gly residues. Results: Using THP-1 cell anti-inflammatory assays for various peptides, studies showed that P1-47-55 was as potent as P1-47-66 and that the N-terminal N47 was essential. In anti-inflammatory activity assays, some P3 peptides shorter than P3-42-65 (e.g., P3-44-54, P3-51-65, and P3-51-59) were as active as P3-42-65. Remarkably, this is the first example for any PAR-agonist peptide that does not require a native-like sequence with an intact free alpha-amino N-terminus to agonize the PAR response. For bivalent P1:P3 studies, we covalently linked P3-47-55 to P3-51-65 using oligo-Gly linkers with 4-14 Gly residues. In anti-inflammatory activity assays, the maximal activity was seen for a 10-Gly linker. The bivalent peptide comprising P1-47-55-[Gly(10 residues)]-P3-51-65, designated as the G10 peptide, was much more potent than any P1 peptide or P3 peptide in anti-inflammatory THP-1 cell assays in dose-response studies. In TEER studies of thrombin-challenged EAhy926 EC where APC markedly attenuates thrombin-induced endothelial barrier dysfunction, P1-47-55 and the G10 peptide mimicked APC's protective actions. In dose-response studies, the G10 peptide was much more potent than the P1-47-55 peptide or any combination of P1-47-55 with a P3 peptide. The P3 peptides alone were very weak. In TEER studies of thrombin-challenged murine aortic EC where murine APC markedly attenuates thrombin-induced endothelial barrier dysfunction, the G10 peptide comprising murine PAR sequences mimicked murine APC's protective actions. Conclusions: Covalent linkage of a PAR1-derived peptide agonist to a PAR3-derived peptide agonist using a 10-Gly-residue linker provided the first heterobivalent PAR peptide GPCR agonist. This heterobivalent G10 peptide potently mimicked APC's cytoprotective activities of anti-inflammation and endothelial barrier stabilization. Molecular mechanisms for G10 likely involve both orthosteric and allosteric effects for PAR1 and PAR3 which can form heterodimers. Given APC's beneficial cell signaling effects in many preclinical injury models and given the increasing usage of peptide-based drugs, studies of the G10 peptide for beneficial APC-mimicking effects may help to identify important directions for future translational research.

A new sulphur-containing metabolite from a mangrove endophytic fungus Aspergillus sp. GXNU-MA

A new sulphur-containing metabolite, asperiguxidione A (1), was isolated from a mangrove endophytic fungus Aspergillus sp. GXNU-MA, and four known alkaloids 2-5, were isolated together from this strain. Their structures were determined by the com­bination of 1D and 2D NMR spectroscopy, HR-ESI-MS, and ECD analysis. Compounds 1 and 2 exhibited mediate activity against Staphylococcus aureus and Enterobacter aerogenes with equal MIC values of 12.5 μg/mL. Compound 3 reduced NO production in LPS-stimulated cells with an IC50 value of 13.329 ± 0.53 μg/mL in the anti-inflammatory assay.

Synthesis, Characterization and in-vitro Biological Studies of Curcumin decorated Biogenic Selenium Nanoparticles

Selenium nanoparticles have drawn interest recently, due to their distinctive physicochemical characteristics and potential usage in numerous applications, including medicine, electronics and catalysis. In this study, we employed Biancaea sappan (BS), often known as sappan wood or Brazilwood, which is a flowering tree that is endemic to Southeast Asia. Due to its antioxidant, anti-inflammatory and anticancer properties, this tree’s heartwood is frequently used in traditional medicine. Additionally, the wood is used to make natural dyes. In this study, we aim to develop an ecofriendly selenium nanoparticle using BS[Formula: see text] as a natural reducing agent and curcumin as a capping agent. The nanoparticles were synthesized using the green synthesis method and characterized using various techniques. Biocompatibility was evaluated using hemolytic assay and the bioactivity of the nanoparticles was assessed using anti-inflammatory and antioxidant assays. Selenium nanoparticles (BCSeN) were successfully synthesized using BS bark extract and functionalized with curcumin. The nanoparticles were characterized by UV-Vis Spectroscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy and Energy-dispersive X-ray spectroscopy (EDX) analysis, confirming their morphology, crystallinity, functionalization, elemental composition, size and stability. In vitro, bioactivity studies revealed that the BCSeN exhibited significant anti-inflammatory activity. They also demonstrated notable antioxidant efficacy against DPPH (2,2-diphenyl-1-picrylhydrazyl) and were also found to have minimal hemolytic potential. Our findings highlight the potential of BCSeN as a promising candidate for anti-inflammatory and antioxidant applications. However, further in-depth analysis is required to fully understand their efficacy and toxicity.

Bioprospecting-based untargeted metabolomics identifies alkaloids as potential anti-inflammatory bioactive markers of Ocotea species (Lauraceae).

Species within the Ocotea genus (Lauraceae), have demonstrated an interesting profile of bioactivities. Renowned for their diverse morphology and intricate specialized metabolite composition, Ocotea species have re-emerged as compelling candidates for bioprospecting in drug discovery research. However, it is a genus insufficiently studied, particularly regarding anti-inflammatory activity. To investigate the anti-inflammatory activity of Ocotea spp. extracts and determine the major markers in this genus. Extracts of 60 different Ocotea spp. were analysed by an ex vivo anti-inflammatory assay in human whole blood. The experiment estimates the prostaglandin E2 levels, which is one of the main mediators of the inflammatory cascade, responsible for the classical symptoms of fever, pain, and other common effects of the inflammatory process. Untargeted metabolomics analysis through liquid chromatography coupled with high-resolution mass spectrometry was performed, along with statistical analysis, to investigate which Ocotea metabolites are correlated with their anti-inflammatory activity. The anti-inflammatory screening indicated that 49 out of 60 Ocotea spp. extracts exhibited significant inhibition of PGE2 release compared to the vehicle (p < 0.05). Furthermore, 10 of these extracts showed statistical similarity to the reference drugs. The bioactive markers were accurately identified using multivariate statistics combined with a fold change (> 1.5) and adjusted false discovery rate analysis as unknown compounds and alkaloids, with a majority of aporphine and benzylisoquinolines. These alkaloids were annotated with an increased level of confidence since MSE spectra were compared with comprehensive databases. This study represents the first bioprospecting report revealing the anti-inflammatory potential of several Ocotea spp. The determination of their anti-inflammatory markers could contribute to drug discovery and the chemical knowledge of the Ocotea genus.