Egg Test-chorioallantoic Membrane Research Articles

Antifungal, Antioxidant, and Irritative Potential of Citronella Oil (Cymbopogon nardus) Associated with Phenethyl Ester of Caffeic Acid (CAPE)

The present study aimed to analyze the antifungal, antioxidant, and irritant potential of citronella oil, both isolated and combined with caffeic acid phenethyl ester (CAPE), for topical oral candidiasis. The antioxidant potential was evaluated using two methods, the DPPH test and the reducing power test (FRAP), while the irritant potential of the solutions was assessed through the hen’s egg chorioallantoic membrane test (HET-CAM). The DPPH test (IC50) values for the CITRO III + CAPE III combination were 32 ± 9 mg/mL, and for isolated CAPE, 13 ± 3 mg/mL. The results from the FRAP method revealed a low iron-reducing power for the combination of 1.25 mg/mL of citronella and 0.0775 mg/mL of CAPE (CITRO III + CAPE III), showing no significant difference compared to the isolated solution of 0.15 mg/mL of CAPE. The antibacterial activity of CAPE and isolated citronella in vitro against microorganisms was evaluated using two methods: microdilution and biofilm assay. The results showed that the MIC and MFC values were 0.5 mg/mL for citronella at both tested times (24 h and 48 h). For CAPE, the MFC values were 0.031 mg/mL. For the biofilm assay, the isolated compounds and combinations at 1 min and 6 h showed significantly different results from the controls (p < 0.05). Furthermore, the HET-CAM results demonstrated the absence of irritability. Based on these premises, the antifungal and antioxidant actions, and absence of irritability were proven. Moreover, this work presents a natural antifungal of interest to the pharmaceutical industry.

Gallic acid loaded self-nano emulsifying hydrogel-based drug delivery system against onychomycosis.

Aim: To developed and investigate gallic acid (GA) loaded self-nanoemulsifying drug delivery systems (SNEDDS) for treating onychomycosis via transungual route.Materials & methods: The SNEDDS were prepared by direct dispersion technique and were evaluated for characteristics parameters using Fourier transform infrared, differential scanning calorimetry, confocal microscopy, transmission electron microscopy and zeta sizer. Furthermore, the safety of prepared formulation was evaluated via Hen's egg test-chorioallantoic membrane study and stability was confirmed using different parameters. Also, its effectiveness was evaluated against fungal strain Trichophyton mentagrophytes.Results: The SNEDDS displayed a particle size of 199.8±4.21nm and a zeta potential; of -22.75±2.09mV. Drug release study illustrated a sustained release pattern with a release of 70.34±0.20% over a period of 24h. The penetration across the nail plate was found to be 1.59±0.002µg/mg and 0.97±0.001µg/mg forGA loaded SNEDDS and GA solution respectively. An irritation score of 0.52±0.005 and 3.84±0.001 was reported for GA loaded SNEDDS hydrogel and GA solution, indicating a decrease in the drug's irritation potential from slightly irritating to nonirritating due to its entrapment within the SNEDDS.Conclusion: GA loaded SNEDDS has potential to address limitations of conventional treatments, enhancing the drug's efficacy and reducing the likelihood of resistance in the treatment of Onychomycosis.

Kaempferia galanga L. alleviates the expression of COX-2 and NF-kappaB-p65 in the oral mucosa ulcer of Wistar rats and exhibits no irritation toward the vascular membrane of chicken eggs and the skin of albino rabbits

The global incidence of recurrent aphthous stomatitis in 2018 reached 5–66 % of the population, while in Indonesia 8 %. Moreover, the prevalence of oral mucosal fibrosis and recurrent aphthous stomatitis among male doctors and nurses in China was 21.24 % and 24.27 %, respectively. Our previous study has shown that the ethanol extract of Kaempferia galanga L. rhizome (EKGR) revealed an accelerated wound-healing effect in the oral mucosa ulcer of Wistar rats. This study aims to explore the effects of EKGR on the expression of NF-kappaB-p65 and COX-2 in the tongue tissue of male Wistar rats by Western blot analysis and immunohistochemistry technique, its safety towards the vascular membrane of the egg chorioallantoic membrane, and its single-dose application on the skin of male rabbits. The rats were randomly assigned into 7 groups: the normal control; the negative control; the positive control (treated with triamcinolone acetonide); and 4 treatment groups of EKGR (0.5 %; 1 %; 2 %; 4 %). Western blot and immunohistochemistry methods were used to measure the expression of NF-kappaB-p65 and COX-2. The hen's egg test-chorioallantoic membrane assay was employed to predict the safety of EKGR towards the vascular membrane. Moreover, the effect of 200 mg/kg BW EKGR application on the dorsal skin of male albino rabbits was also evaluated. EKGR inhibits the expression of NF-kappaB-p65 and COX-2 as proven by WB and IHC results. In the HET-CAM assay, all concentrations of EKGR do not induce irritation responses, which elicits the safety of EKGR. The administration of EKGR causes mild irritation to the dorsal skin of male rabbits but does not induce erythema and edema, no significant changes in BW, no toxic effects on organ macroscopic examination or histopathology, and does not induce abnormalities in the hematological profile of male albino rabbits. EKGR has confirmed its anti-inflammatory activity by suppressing the expression of COX-2 and NF-kappaB-p65 in the oral mucosa ulcer of Wistar rats. EKGR is safe as it does not exhibit irritating potential and harmful effects.

Antioxidant Extracts from Greek and Spanish Olive Leaves: Antimicrobial, Anticancer and Antiangiogenic Effects.

Olea europaea L. is the most valuable species of the Olea type, and its products offer a wide range of therapeutical uses. The olive tree has been extensively studied for its nourishing qualities, and the "Mediterranean diet", which includes virgin olive oil as a key dietary component, is strongly associated with a reduced risk of cardiovascular disease and various malignancies. Olive leaves, a by-product in the olive harvesting process, are valued as a resource for developing novel phytomedicines. For this purpose, two ethanolic extracts obtained from Olivae folium from Spain (OFS) and Greece (OFG) were investigated. Our findings contribute to a wider characterization of olive leaves. Both extracts displayed important amounts of phenolic compounds and pentacyclic triterpenes, OFG having higher concentrations of both polyphenols, such as oleuropein and lutein, as well as triterpenes, such as oleanolic acid and maslinic acid. The antioxidant capacity is similar for the two extracts, albeit slightly higher for OFG, possibly due to metal polyphenol complexes with antioxidant activity. The extracts elicited an antimicrobial effect at higher doses, especially against Gram-positive bacteria, such as Streptococcus pyogenes. The extract with lower inorganic content and higher content of polyphenols and triterpenic acids induced a strong anti-radical capacity, a selective cytotoxic effect, as well as antimigratory potential on A375 melanoma cells and antiangiogenic potential on the CAM. No irritability and a good tolerability were noted after evaluating the extracts on the in vivo Hen's Egg Test-Chorioallantoic Membrane (HET-CAM). Therefore, the present data are suggestive for the possible use of the two types of olive leaf products as high-antioxidant extracts, potentially impacting the healthcare system through their use as antimicrobial agents and as anticancer and anti-invasion treatments for melanoma.

Enhanced Antioxidant, Anti-Aging, Anti-Tyrosinase, and Anti-Inflammatory Properties of Vanda coerulea Griff. Ex Lindl. Protocorm through Elicitations with Chitosan.

This study aimed to investigate the effects of elicitors on Vanda coerulea Griff. Ex Lindl. protocorms to enhance bioactive compound production and evaluate their biological activities relevant to cosmeceutical applications. The protocorms were developed from the callus treated with different elicitors, including 6-benzylaminopurine (BA), methyl jasmonate (MeJA), and chitosan. Both the adult plant and protocorms were extracted by maceration in 80% methanol and investigated for their chemical compositions using high-performance liquid chromatography. The extracts were evaluated for antioxidant, anti-collagenase, anti-elastase, and anti-tyrosinase activities. In addition, anti-inflammatory properties were assessed using a real-time polymerase chain reaction. The irritation potency was evaluated using the hen's egg test-chorioallantoic membrane test. The findings revealed that protocorms treated with BA and chitosan developed a greener color, while those treated with MeJA exhibited a distinct darker coloration. Elicitation with BA and chitosan resulted in protocorms with comparable or higher levels of syringic acid, rutin, and quercin compared with the adult plant, with rutin being the most prominent identified compound. Furthermore, rutin was reported as the compound responsible for all biological activities. The chitosan-treated protocorm extract exhibited potent inhibition against oxidation, collagenase, elastase, tyrosinase, and inflammatory cytokines, along with a nonirritating effect, making it a promising candidate for cosmeceutical applications.

In vitro and in vivo studies of ocular topically administered NLC for the treatment of uveal melanoma

Uveal melanoma is one of the most common and aggressive intraocular malignancies, and, due to its great capability of metastasize, it constitutes the most incident intraocular tumor in adults. However, to date there is no effective treatment since achieving the inner ocular tissues still constitutes one of the greatest challenges in actual medicine, because of the complex structure and barriers. Uncoated and PEGylated nanostructured lipid carriers were developed to achieve physico-chemical properties (mean particle size, homogeneity, zeta potential, pH and osmolality) compatible for the ophthalmic administration of (S)-(–)-MRJF22, a new custom-synthetized prodrug for the potential treatment of uveal melanoma. The colloidal physical stability was investigated at different temperatures by Turbiscan® Ageing Station. Morphology analysis and mucoadhesive studies highlighted the presence of small particles suitable to be topically administered on the ocular surface. In vitro release studies performed using Franz diffusion cells demonstrated that the systems were able to provide a slow and prolonged prodrug release. In vitro cytotoxicity test on Human Corneal Epithelium and Human Uveal Melanoma cell lines and Hen’s egg-chorioallantoic membrane test showed a dose-dependent cytotoxic effect of the free prodrug on corneal cells, whose cytocompatibility improved when encapsulated into nanoparticles, as also confirmed by in vivo studies on New Zealand albino rabbits. Antiangiogenic capability and preventive anti-inflammatory properties were also investigated on embryonated eggs and rabbits, respectively. Furthermore, preliminary in vivo biodistribution images of fluorescent nanoparticles after topical instillation in rabbits’ eyes, suggested their ability to reach the posterior segment of the eye, as a promising strategy for the treatment of choroidal uveal melanoma.

Potential of Bioactive Protein and Protein Hydrolysate from Apis mellifera Larvae as Cosmeceutical Active Ingredients for Anti-Skin Aging.

This study aimed to extract bioactive proteins and protein hydrolysates from Apis mellifera larvae and assess their potential application in cosmetics as well as their irritation properties. The larvae were defatted and extracted using various mediums, including DI water, along with 0.5 M aqueous solutions of sodium hydroxide, ascorbic acid, citric acid, and hydrochloric acid. Subsequently, the crude proteins were hydrolyzed using the Alcalase® enzyme. All extracts underwent testing for antioxidant activities via the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) and Griess assays. Anti-aging properties were evaluated in terms of anti-collagenase and anti-hyaluronidase effects. Irritation potential was assessed using the hen's egg chorioallantoic membrane (HET-CAM) test. The results revealed that the sodium hydroxide extraction showed promising outcomes in terms of yield, protein content, and effectiveness in inhibiting hyaluronidase, with the highest inhibition at 78.1 ± 1.5%, comparable to that of oleanolic acid. Conversely, crude protein extracted with ascorbic acid and its hydrolysate showed notable antioxidant and collagenase-inhibitory activities. Remarkably, their anti-collagenase effects were comparable to those of ascorbic acid and lysine. Additionally, it demonstrated safety upon testing with the CAM. In conclusion, the findings provided valuable insights into the utilization of A. mellifera larval proteins as active ingredients with a wide range of cosmeceutical applications, particularly due to their antioxidant, anti-aging, and low irritation properties, which hold significant promise for anti-skin wrinkles.

Cytisine eye drops for benzalkonium chloride-induced dry eye: safety and efficacy evaluation

This experiment aimed to investigate the feasibility of cytisine (CYT) in treating eye diseases with ocular topical application. An in vitro cytotoxicity test, a hen’s egg test-chorioallantoic membrane (HET-CAM), and a mouse eye tolerance test were used to fully reveal the ocular safety profiles of CYT. For the efficacy evaluations, CYT’s effects on cell wound healing, against H2O2-induced oxidative stress damages on cells, and on benzalkonium chloride (BAC)–induced dry eye disease (DED) in mice were evaluated. Results showed that CYT did not show any cytotoxicities at concentrations no higher than 250 μg/ml, while lipoic acid (α-LA) at 250 μg/ml and BAC at 1.25 μg/ml showed significant cytotoxicities within 48 h incubation. The HET-CAM and mouse eye tolerance test confirmed that 0.5% CYT eye drops demonstrated good safety characteristics. Efficacy evaluations showed that CTY significantly promoted cell migration and wound healing. CYT significantly improved cell survival against H2O2-induced oxidative stress damage by reversing the imbalance between the reactive oxygen species (ROS) and antioxidant defense mechanisms. The animal evaluation of the BAC-induced dry eye model revealed that CYT demonstrated a strong treatment effect, including reversing ocular surface damages, recovering corneal sensitivity, and inhibiting neovascularization; HMGB1/NF-κB signaling was involved in this DED treatment by CTY. In conclusion, CYT had strong experimental treatment efficacy against DED with good ocular safety profiles, and it might be a novel and promising drug for DED.

Glycol Chitosan-Poly(lactic acid) Conjugate Nanoparticles Encapsulating Ciprofloxacin: A Mucoadhesive, Antiquorum-Sensing, and Biofilm-Disrupting Treatment Modality for Bacterial Keratitis.

Bacterial keratitis (BK) causes visual morbidity/blindness if not treated effectively. Here, ciprofloxacin (CIP)-loaded nanoparticles (NPs) using glycol chitosan (GC) and poly(lactic acid) (PLA) conjugate at three different ratios (CIP@GC(PLA) NPs (1:1,5,15)) were fabricated. CIP@GC(PLA) NPs (1:1) were more effective than other tested ratios, indicating the importance of optimal hydrophobic/hydrophilic balance for corneal penetration and preventing bacterial invasion. The CIP@GC(PLA) (NPs) (1:1) realized the highest association with human corneal epithelial cells, which were nonirritant to the hen's egg-chorioallantoic membrane test (HET-CAM test) and demonstrated significant antibacterial response in the in vitro minimum inhibitory, bactericidal, live-dead cells, zone of inhibition, and biofilm inhibition assays against the keratitis-inducing pathogen Pseudomonas aeruginosa. The antiquorum sensing activity of GC has been explored for the first time. The NPs disrupted the bacterial quorum sensing by inhibiting the production of virulence factors, including acyl homoserine lactones, pyocyanin, and motility, and caused significant downregulation of quorum sensing associated genes. In the in vivo studies, CIP@GC(PLA) NPs (1:1) displayed ocular retention in vivo (∼6 h) and decreased the opacity and the bacterial load effectively. Overall, the CIP@GC(PLA) NP (1:1) is a biofilm-disrupting antiquorum sensing treatment regimen with clinical translation potential in BK.

Development of pH-Sensitive Nanoparticle Incorporated into Dissolving Microarray Patch for Selective Delivery of Methotrexate.

Rheumatoid arthritis (RA) is a systemic autoimmune disease that attacks human joints. Methotrexate (MTX), as one the most effective medications to treat RA, has limitations when administered either orally or by injection. To overcome this limitation, we formulated MTX through a smart nanoparticle (SNP) combined with dissolving microarray patch (DMAP) to achieve selective-targeted delivery of RA. SNP was made using the combination of polyethylene glycol (PEG) and polycaprolactone (PCL) polymers, while DMAP was made using the combination of hyaluronic acid and polyvinylpyrrolidone K-30. SNP-DMAP was then evaluated for its mechanical and chemical characteristics, ex vivo permeation test, in vivo pharmacokinetic study, hemolysis, and hen's egg test-chorioallantoic membrane (HET-CAM) test. The results showed that the characteristics of the SNP-DMAP-MTX formulas meet the requirements for transdermal delivery, with the particle size of 189.09 ±12.30 nm and absorption efficiency of 65.40 ± 5.0%. The hemolysis and HET-CAM testing indicate that this formula was non-toxic and non-irritating. Ex vivo permeation shows a concentration of 51.50 ± 3.20 µg/mL of SNP-DMAP-MTX in PBS pH 5.0. The pharmacokinetic profile of SNP-DMAP-MTX showed selectivity and sustained release compared with oral and DMAP-MTX with values of t1/2 (4.88 ± 0 h), Tmax (8 ± 0 h), Cmax (0.50 ± 0.04 μg/mL), AUC (3.15 ± 0.54 μg/mL.h), and mean residence time (MRT) (9.13 ± 0 h). The developed SNP-DMAP-MTX has been proven to deliver MTX transdermal and selectively at the RA site, potentially avoiding conventional MTX side effects and enhancing the effectiveness of RA therapy.

The potential of Paeonia lactiflora pall seeds oil as a pure natural cosmetics raw material: InVitro findings.

As a traditional Chinese herbal medicine, Paeonia lactiflora Pall is rich in various active ingredients such as polysaccharides and total flavonoids while having ornamental value. It has potential application value in the development of food and cosmetics. To study the invitro efficacy of Paeonia lactiflora Pall seeds oil. Firstly, the levels of linolenic acid and linoleic acid in Paeonia lactiflora Pall seeds oil were quantified using gas chromatography. The impact of Paeonia lactiflora Pall seeds oil on the proliferation rate of B16F10 cells was assessed through the CCK-8 method, while the melanin content of B16F10 cells was determined using the sodium hydroxide lysis method. The inhibitory effects of Paeonia lactiflora Pall seeds oil on elastase, collagenase and hyaluronidase were evaluated by biochemical techniques invitro. Lastly, the hen's egg chorioallantoic membrane test (HET-CAM) was conducted to confirm the absence of eye irritation caused by Paeonia lactiflora Pall seeds oil. Paeonia lactiflora Pall seeds oil within a certain volume concentration range (0.5%-4%) had no effect on the proliferation of B16F10 cells. Paeonia lactiflora Pall seeds oil showed significant inhibition of elastase, collagenase and hyaluronidase. Notably, the highest concentration tested, 4% Paeonia lactiflora Pall seed oil, yielded the most pronounced outcomes without causing any irritation. A certain concentration of Paeonia lactiflora Pall seeds oil has a significant effect on decreasing the melanin content in B16F10 cells and inhibiting the activities of elastase, collagenase, and hyaluronidase, which can provide a reference for the development of pure natural cosmetics raw materials.

Advancing ocular safety research: A comprehensive examination of benzocaine acute exposure without animal testing

Benzocaine is a widely employed local anaesthetic; however, there is a notable dearth of preclinical and clinical evidence regarding its safety in ophthalmological products. To address this, a comprehensive strategy incorporating in silico and in vitro methodologies was proposed for assessing benzocaine's ocular toxicity without animal testing. To collect the in silico evidence, the QSAR Toolbox (v4.5) was used. A single exposure to two benzocaine concentrations (2% and 20%) was evaluated by in vitro methods. Hen’s Egg Chorioallantoic Membrane Test (HET-CAM) was performed to evaluate the effects on the conjunctiva. To study corneal integrity, Short Time Exposure test (STE) and Bovine Corneal Opacity and Permeability (BCOP) assay, followed by histopathological analysis, were carried out. Results from both in silico and in vitro methodologies categorize benzocaine as non-irritating. The histopathological analysis further affirms the safety of using benzocaine in eye drops, as no alterations were observed in evaluated corneal strata. This research proposes a useful combined strategy to provide evidence on the safety of local anaesthetics and particularly show that 2% and 20% benzocaine solutions do not induce eye irritation or corneal damage, supporting the potential use of benzocaine in the development of ophthalmic anesthetic products.

Unlocking the beauty benefits: Exploring biosurfactants from Scheffersomyces shehatae for cosmetics

AbstractAs the cosmetic market continues to transform, innovative ingredients such as biosurfactants (BS) emerge as a fundamental step towards a generation of cosmetic products characterized by efficacy, safety, and environmental responsibility. In alignment with the global push towards natural, renewable, and non‐toxic materials, BS stands out as an interesting alternative to conventional synthetic surfactants. BS are a type of green surfactants and compared to synthetic surfactants derived from petroleum they are more biodegradable and have low or no toxicity. This work aimed to investigate the suitability of a biosurfactant extracted from the yeast Scheffersomyces shehatae (BS‐SS) to be used in cosmetic applications. The cytotoxicity of the BS‐SS was tested in fibroblast (L929) and keratinocyte (HaCaT) cell lines and the potential for ocular irritation was assessed by the hen's egg chorioallantoic membrane test (HET‐CAM). The cleaning efficiency of the BS‐SS was measured as the ability of make‐up removal in pig skin. Our results showed that the IC 50 values were 10.49 mg/mL for fibroblasts and 11.77 mg/mL for keratinocytes, and the biosurfactant did not result in hemorrhage, lysis, and coagulation damages in the chorioallantoic membrane, which classifies the biosurfactant solution as non‐irritant. Therefore, ocular toxicity and in vitro skin cell toxicity results showed that BS‐SS can be tolerable and safe for cosmetic purposes. BS‐SS featured promising cleaning properties, similar to the cleansing activity of the 1% SDS solution and of micellar water for lipstick and foundation products. In conclusion, the results indicate its potential use in cosmetic applications.

Antimicrobial Properties Related to Anti-Acne and Deodorant Efficacy of Hedychium coronarium J. Koenig Extracts from Pulsed Electric Field Extraction.

This study investigated the potential of pulsed electric field (PEF) extraction in enhancing the antimicrobial properties related to anti-acne and deodorant properties of Hedychium coronarium extract. The dried leaf and rhizome of H. coronarium were extracted using 95% v/v ethanol through both conventional solvent extraction and PEF extraction techniques (10, 14, and 20 kV/cm). The chemical composition of the extracts was analyzed. The antimicrobial activities, specifically in relation to acne treatment against Cutibacterium acnes and deodorant properties against Staphylococcus aureus, Bacillus subtilis, Micrococcus luteus, Pseudomonas aeruginosa, and Escherichia coli, were determined. The irritation profile of was evaluated using the hen's egg chorioallantoic membrane test. The results showed that PEF extraction increased the extract yield, particularly at an electric field strength of 20 kV/cm. Furthermore, PEF extraction significantly enhanced the ellagic acid content, particularly in the leaf extract. Furthermore, the leaf extract demonstrated stronger inhibitory effects against microorganisms associated with body odor and acne compared to the rhizome extract. Notably, all extracts exhibited no signs of irritation, indicating their safety. Overall, the findings suggest that PEF extraction from H. coronarium enhances yield, bioactive compound content, and antimicrobial effects. This indicates the potential of the extract for acne treatment and deodorant use.

Exploring the Ocular Absorption Pathway of Fasudil Hydrochloride towards Developing a Nanoparticulate Formulation with Improved Performance.

Rho-kinase (ROCK) inhibitors represent a new category of anti-glaucoma medications. Among them, Fasudil hydrochloride, a selective ROCK inhibitor, has demonstrated promising outcomes in glaucoma treatment. It works by inhibiting the ROCK pathway, which plays a crucial role in regulating the trabecular meshwork and canal of Schlemm's aqueous humor outflow. This study aims to investigate the ocular absorption pathway of Fasudil hydrochloride and, subsequently, develop a nanoparticle-based delivery system for enhanced corneal absorption. Employing the ionic gelation method and statistical experimental design, the factors influencing chitosan nanoparticle (Cs NP) characteristics and performance were explored. Fasudil in vitro release and ex vivo permeation studies were performed, and Cs NP ocular tolerability and cytotoxicity on human lens epithelial cells were evaluated. Permeation studies on excised bovine eyes revealed significantly higher Fasudil permeation through the sclera compared to the cornea (370.0 μg/cm2 vs. 96.8 μg/cm2, respectively). The nanoparticle size (144.0 ± 15.6 nm to 835.9 ± 23.4 nm) and entrapment efficiency range achieved (17.2% to 41.4%) were predominantly influenced by chitosan quantity. Cs NPs showed a substantial improvement in the permeation of Fasudil via the cornea, along with slower release compared to the Fasudil aqueous solution. The results from the Hen's Egg Test Chorioallantoic Membrane (HET-CAM) and Bovine Corneal Opacity and Permeability (BCOP) tests indicated good conjunctival and corneal biocompatibility of the formulated chitosan nanoparticles, respectively. Lens epithelial cells displayed excellent tolerance to low concentrations of these nanoparticles (>94% cell viability). To the best of our knowledge, this is the first report on the ocular absorption pathway of topically applied Fasudil hydrochloride where the cornea has been identified as a potential barrier that could be overcome using Cs NPs.

Phytochemical Compositions and Cosmeceutical Activities of Rosa damascena Mill. Leaf Extracts from Environmentally Friendly Extraction Technique

A significant amount of Rosa damascena Mill. leaves were discarded as waste during the harvesting and pruning for flowering. The current study aimed to investigate the potential of using these agricultural waste products in the cosmetics industry. Dried R. damascena leaves were extracted using maceration or environmentally friendly extractions, including infusion, digestion, ultrasonic, microwave, micellar, and pulsed electric fields (PEF) extraction. The extracts were analyzed for their chemical compositions and assessed for antioxidant, anti-tyrosinase, and anti-aging properties. The irritation profile of each extract was investigated by the hen’s egg-chorioallantoic membrane (HET-CAM) test. The results noted that extraction by ethanol yielded significantly higher extract content than deionized water (P < 0.05). However, total phenolic and flavonoid content was found to be greater, whereas the bioactive compounds, including rutin and rosmarinic acid, were only found in the aqueous extracts. On the other hand, kojic acid was found in the extract from infusion and maceration. Interestingly, the extracts from micellar and PEF extractions were found to significantly scavenge the radical, which were related to their rutin contents. Their collagenase inhibition (69.1 ± 10.1% and 54.8 ± 19.1%, respectively) were equivalent to those of epigallocatechin gallate, a well-known anti-aging compound (76.0 ± 1.2%). Besides, all extracts were safe since they induced no irritation in the HET-CAM test. In conclusion, environmentally friendly extraction of bioactive components from R. damascena leaves were suggested, with the potential to be exploited as an anti-aging and whitening ingredient for further use in cosmeceutical area. Keywords: Rosa damascene, Green extraction, Antioxidant, Anti-tyrosinase, Collagen, Elastin; Hyaluronan, Irritation

Protective Effect of Panax notoginseng Extract Fermented by Four Different Saccharomyces cerevisiae Strains on H2O2 Induced Oxidative Stress in Skin Fibroblasts.

To produce Panax notoginseng extract as a cosmetic ingredient through Saccharomyces cerevisiae fermentation. We first compared the total sugar content, polysaccharide content, reducing sugar content, total phenolic content, total saponin content, DPPH free radical, ABTS free radical, hydroxyl free radical scavenging ability and ferric reducing antioxidant power (FRAP) of Panax notoginseng fermented extract (pnFE) and unfermented extract (pnWE). Their potential correlations were analyzed by Pearson's correlation analysis. Then, the oxidative stress model of H2O2-induced MSFs was used to evaluate the effects of different pnFE on MSF viability, reactive oxygen species (ROS), malondialdehyde (MDA), and the activities of catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) to explore their protective effects on MSFs subjected to H2O2-induced cellular oxidative damage. Finally, their safety and stability were evaluated by using the red blood cell (RBC) test and hen's egg test-chorioallantoic membrane (HET-CAM) assay, and changes in pH and content of soluble solids, respectively. Compared with pnWE, pnFE has more active substances and stronger antioxidant capacity. In addition, pnFE has a protective effect on H2O2-induced oxidative stress in MSFs with appropriate safety and stability. PnFE has broad application prospects in the field of cosmetics.

Fluocinolone Acetonide Loaded Chitosan Nanofiber Scaffolds for Treatment of Ocular Disorders: In Vitro Characterization, Ex-Vivo Corneal and Ex-Vivo Scleral Evaluation

Purpose Drugs administered in the ocular region need to overcome ocular barriers without permanently damaging the ocular tissues. Moreover, ocular disorders of the posterior segment are more difficult to treat due to invasive procedures required to reach the posterior segment. Hence, to treat posterior disorders of the eye an attempt was made to develop nanofiber (NF) scaffolds for effective management of chronic posterior uveitis. Nanofibers (NFs) were formulated using the electrospinning technique. Methods NF scaffolds were formulated using the electrospinning technique. The effect of different concentrations of chitosan on NF production was studied by considering different ratios of chitosan (CS) and polyvinyl alcohol (PVA). Physicochemical characterization of NFs was performed to evaluate developed NFs. Results The optimized NF scaffold had a diameter of 129 ± 3 nm. NF scaffolds were found to have a tensile strength of 0.2882 ± 0.078 N/m2, thickness of 0.16 ± 0.05 mm, and drug entrapment of 95 ± 2.0%. The bioadhesive strength of the NF was found to be 257.3 ± 0.04 g/cm2 indicating high bioadhesion of NFs to the ocular tissues. The in-vitro, ex-vivo corneal and ex-vivo scleral drug release after 12 h was found to be 78.4 ± 1.0%, 65.33 ± 0.2% and 78.41 ± 1.0%, respectively. Ex-vivo whole eye model experiment indicated a concentration of about 40 ± 1.75% of drug permeated from corneal layer to the vitreous humor after 12 h. The Hen’s egg test-chorioallantoic membrane study (HET-CAM) study and in-vitro cytotoxicity study on Statens Seruminstitut Rabbit Cornea (SIRC) cell lines indicated that the developed drug-loaded NF scaffolds were found to be non-toxic as compared to pure drug, thus suggesting cytocompatibility. Conclusion Results of HET-CAM, sterility and ex-vivo studies indicate that the developed formulation is non-toxic, sterile, and effective for the ocular delivery of fluocinolone acetonide to the posterior segment of eye.

Promising Anti-Wrinkle Applications of Aromatic Extracts of Hedychium coronarium J. Koenig via Antioxidation and Collagenase Inhibition.

This study aimed to extract aromatic compounds from the rhizomes, leaf sheaths, and leaves of Hedychium coronarium and investigate their chemical compositions, cosmetic/cosmeceutical activities, and irritation potency. The chemical compositions were investigated via gas chromatography-mass spectrometry. The antioxidant activities were evaluated via spectrophotometry. The anti-skin wrinkle properties were investigated via collagenase, elastase, and hyaluronidase inhibition. The irritation potency was observed via a hen's egg-chorioallantoic membrane test. Eucalyptol was detected as a major component in the rhizomes and leaf sheaths, while β-caryophyllene was predominant in the leaves. The absolutes from the rhizomes were the strongest antioxidants, with ABTS scavenging properties similar to L-ascorbic acid. Interestingly, the equivalent concentration (EC1) of the absolute from the rhizome was 0.82 ± 0.01 µg FeSO4/g extract, which was significantly more potent than L-ascorbic acid (0.43 ± 0.03 µg FeSO4/g extract). The rhizome-derived absolute was the most effective against collagenase, while the concretes from the rhizomes and leaf sheaths showed promising anti-hyaluronidase activity with inhibitions of 90.5 ± 1.6% and 87.4 ± 5.1%, respectively. The irritability of the aromatic extracts was not different from that of the vehicle control, proving their safety. Therefore, the Hedychium coronarium rhizome-derived absolute was an attractive and potent antioxidant with anti-collagenase activities, indicating its potential for use in anti-aging formulations.

Ocular antibacterial chitosan-maleic acid hydrogels: In vitro and in vivo studies for a promising approach with enhanced mucoadhesion

The aim was to design and evaluate a chitosan-based conjugate providing high mucoadhesiveness and antibacterial activity for ocular infections treatment. Chitosan was conjugated with maleic acid via amide bond formation and infrared spectroscopy. Furthermore, 2,4,6-Trinitrobenzene sulfonic acid (TNBS) allowed characterization and quantification of conjugated groups, respectively. Biocompatibility was tested via hemolysis assay and Hen's Egg-Chorioallantoic membrane test. Characterization of the pH and osmolarity of hydrogels was followed by mucoadhesion assessment utilizing rheology. In addition, antibacterial studies were carried out towards Escherichia coli by broth microdilution test and agar-disk diffusion assay. In vivo studies were carried out following the already established Draize test and determining pharmacokinetic profile of dexamethasone in aqueous humour. The conjugate exhibited a degree of modification of 50.05 % and no toxicity or irritability. Moreover, mucoadhesive properties were enhanced in 2.68-fold and 1.81-fold for elastic and viscous modulus, respectively. Furthermore, rheological synergism revealed the presence of a gel-like structure. Additionally, broth microdilution and agar disk diffusion studies exhibited enhancement in antibacterial activity. Finally, in vivo studies manifested that hydrogels were highly tolerated, evidencing promising characteristics of the developed conjugate. The conjugate presented promising antimicrobial, long lasting mucoadhesive features and highly improved pharmacokinetics, leading to a revolutionizing approach in the treatment of ocular bacterial infections.