MICROEMULSION-BASED HYDROGEL FOR TOPICAL DELIVERY OF INDOMETHACIN

Objective: The present study aims to develop and characterize microemulsion from herbal infused oil of Zingiber cassumunar (HO) and microemulsion-based hydrogel (MBH) containing indomethacin. The release patterns of indomethacin from MBH were also investigated.Methods: HO was produced by hot extraction of Z. cassumunar rhizome in coconut oil, and characterized for acid value, iodine value, and saponification value. The cytotoxicity of HO on human peripheral blood mononucleared cells (PBMCs) was also investigated. Pseudoternary phase diagram was constructed to study suitable compositions of microemulsion containing HO, oleic acid, Triton X-114, propan-2-ol, and water. Indomethacin was then incorporated into the microemulsion and finally blended with gel base (2% Carbopol 940 or 3% sodium carboxymethylcellulose) to produce MBH. The indomethacin MBH was characterized for appearance, pH, viscosity, and in vitro release characteristics.Results: HO exhibited an acid value of 0.203 ± 0.004 mg of KOH/g, iodine value of 7.39 ± 0.15 g of I2/100 g, and saponification value of 265.4 ± 7.3 mg of KOH/g with no cytotoxic effect on human PBMCs. The microemulsion region in the pseudoternary phase diagram of HO, oleic acid, Triton X-114, propan-2-ol, and water was 45.25%. Six microemulsions (ME1– ME6) containing 10% of HO and oleic acid mixture (1:1) as the oil phase and Triton X-114 and propan-2-ol (3:2) as surfactant mixture were formulated and characterized. The droplet size was in the range of 26 to 32 nm with polydispersity index less than 0.3. They showed a Newtonian flow behavior with the viscosity ranging from 15.12 ± 0.15 to 16.78 ± 0.12 Pas. The microemulsion was incorporated into hydrogel using 3% sodium carboxymethylcellulose or 2% Carbopol 940. Only ME1 – ME3 gave clear MBH; therefore, they were studied in the in vitro release of indomethacin, and the results indicated the sustained-release characteristic fitted the Higuchi model.Conclusion: The topical MBH, containing microemulsion of HO, oleic acid, Triton X-114, propan-2-ol and water, might be a promising approach for sustained transdermal delivery of poor water-soluble compounds, including indomethacin.Â

BIOLOGICAL ACTIVITIES AND CHARACTERIZATION OF THE POD EXTRACTS FROM SOMPOI (ACACIA CONCINNA LINN) GROWN IN NORTHERN THAILAND

Objective: Sompoi or Shikakai (Acacia concinna Linn.) pod extracts were produced and investigated for antioxidant and antityrosinase activities together with the determination of their total phenolic and flavonoid content. In addition, phytochemical screening, cytotoxic effect and HPLC-fingerprint of the extract were also performed.Methods: The pods of A. concinna were extracted using three different solvents including 95%ethanol, 50%ethanol and deionized water and named as ethanolic extract (ET), freeze-dried hydroethanolic extract (HEF), Spray-dried hydroethanolic extract (HES), freeze-dried aqueous extract (HF), and spray-dried aqueous extract (HS). Antioxidant activity of the extracts were determined by three different assays including 1-Diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging assay, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS)-radical scavenging assay and linoleic acid peroxidation assay. Total phenolic content in each extract was evaluated using folin-ciocalteu spectrophotometric assay and the total flavonoid content was also determined using aluminium chloride colorimetric assay. Cytotoxic effect on human peripheral blood mononuclear cells (PBMCs) was investigated using MTT assay. HPLC-fingerprint of the selected extract was finally analysed.Results: HES showed the greatest radical scavenging activity on DPPH and ABTS assays with IC50 of 0.8280±0.0131 mg/ml and 0.1418±0.0012 mg/ml, respectively. Moreover, it showed the highest inhibition on lipid peroxidation with IC50 of 1.8751±0.0307 mg/ml. Among five extracts, HES contained the highest amount of gallic acid and rutin expressed as gallic acid equivalent value (GAE) of 100.51±0.06 mg/g and rutin equivalent value (RE) of 14.67±0.01 mg/g. Therefore, its highest antioxidant activity was due to its highest phenolic and flavonoid content. The %viability of HES-treated PBMCs was comparable to ascorbic acid at each individual concentration. The HPLC-fingerprint of HES, detected at the wavelength of 280 nm, showed one major peak at 9.88 min that can be used as a marker of the HES.Conclusions: A. concinna pod extracts showed good in vitro antioxidant and antityrosinase activities with moderate total phenolic and flavonoid content. HES showed the greatest activity with safety profile comparable to ascorbic acid. It might be a promising approach for application in the cosmetic or cosmeceutical products.Â

The in vitro effects of black soldier fly larvae (Hermitia illucens) oil as a high-functional active ingredient for inhibiting hyaluronidase, anti-oxidation benefits, whitening, and UVB protection

Objective: Larvae of Hermitia illucens, or black soldier fly larvae (BSFL), have been recognized for their high lipid yield with a remarkable fatty acid profile. BSFL oil (SFO) offers the added value of a low environmental footprint and a sustainable product. In this study, the characteristics and cosmetic-related activities of SFO were investigated and compared with rice bran oil, olive oil and krill oil which are commonly used in cosmetics and supplements.Methods: The physicochemical characteristics were determined including acid value, saponification value, unsaponifiable matter and water content of SFO. The fatty acid composition was determined using GC-MS equipped with TR-FAME. The in vitro antioxidant properties were determined using DPPH, FRAP and lipid peroxidation inhibition assays. Antihyaluronidase (anti-HAase) activity was measured by detecting enzyme activity and molecular docking of candidate compounds toward the HAase enzyme. The safety assessment towards normal human cells was determined using the MTT assay and the UVB protection upon UVB-irradiated fibroblasts was determined using the DCF-DA assay. The whitening effect of SFO was determined using melanin content inhibition.Results: SFO contains more than 60% polyunsaturated fatty acids followed by saturated fatty acids (up to 37%). The most abundant component found in SFO was linoleic acid (C18:2 n-6 cis). Multiple anti-oxidant mechanisms of SFO were discovered. In addition, SFO and krill oil prevented hyaluronic acid (HA) degradation via strong HAase inhibition comparable with the positive control, oleanolic acid. The molecular docking confirmed the binding interactions and molecular recognition of major free fatty acids toward HAase. Furthermore, SFO exhibited no cytotoxicity on primary human skin fibroblasts, HaCaT keratinocytes and PBMCs (IC50 values > 200 μg/mL). SFO possessed significant in-situ anti-oxidant activity in UVB-irradiated fibroblasts and the melanin inhibition activity as effective as well-known anti-pigmenting compounds (kojic acid and arbutin, p < 0.05).Conclusion: This study provides scientific support for various aspects of SFO. SFO can be considered an alternative oil ingredient in cosmetic products with potential implications for anti-skin aging, whitening and UVB protection properties, making it a potential candidate oil in the cosmetic industry

Chemical Constituents, Antioxidant, Anti-MMPs, and Anti-Hyaluronidase Activities of Thunbergia laurifolia Lindl. Leaf Extracts for Skin Aging and Skin Damage Prevention

This study aimed to investigate the potential usage of Thunbergia laurifolia Lindl. leaf extracts in the cosmetic industry. Matrix metalloproteinases (MMPs) and hyaluronidase inhibition of T. laurifolia leaf extracts, prepared using reflux extraction with deionized water (RE) and 80% v/v ethanol using Soxhlet’s apparatus (SE), were determined. Rosmarinic acid, phenolics, and flavonoids contents were determined using high-performance liquid chromatography, Folin–Ciocalteu, and aluminum chloride colorimetric assays, respectively. Antioxidant activities were determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and linoleic acid-thiocyanate assays. MMP-1 inhibition was investigated using enzymatic and fluorescent reactions, whereas MMP-2, MMP-9, and hyaluronidase inhibition were investigated using gel electrophoresis. Cytotoxicity on human fibroblast cell line was also investigated. The results demonstrated that SE contained significantly higher content of rosmarinic acid (5.62% ± 0.01%) and flavonoids (417 ± 25 mg of quercetin/g of extract) but RE contained a significantly higher phenolics content (181 ± 1 mg of gallic acid/g of extract; p < 0.001). SE possessed higher lipid peroxidation inhibition but less DPPH• scavenging activity than RE. Both extracts possessed comparable hyaluronidase inhibition. SE was as potent an MMP-1 inhibitor as gallic acid (half maximal inhibitory concentration values were 12.0 ± 0.3 and 8.9 ± 0.4 mg/cm3, respectively). SE showed significantly higher MMP-2 and MMP-9 inhibition than RE (p < 0.05). Therefore, SE is a promising natural anti-ageing ingredient rich in rosmarinic acid and flavonoids with antioxidant, anti-hyaluronidase, and potent MMPs inhibitory effects that could be applied in the cosmetic industry

Enhanced Antioxidant, Hyaluronidase, and Collagenase Inhibitory Activities of Glutinous Rice Husk Extract by Aqueous Enzymatic Extraction

In this research, we aimed to compare the biological activities related to cosmeceutical applications of glutinous rice husk extracted by aqueous enzymatic extraction (AEE) and conventional solvent extraction. Cellulase enzymes were used to assist the extraction process. The vanillic and ferulic acid contents of each extract were investigated by high-performance liquid chromatography, and their antioxidant and anti-aging activities were investigated by spectrophotometric methods. The irritation effects of each extract were investigated by the hen’s egg test on chorioallantoic membrane. The rice husk extract from AEE using 0.5% w/w of cellulase (CE0.5) contained the significantly highest content of vanillic and ferulic acid (p v/v ethanol was used. Therefore, AEE is suggested as a green extraction method that can be used instead of the traditional solvent extraction technique given its higher yield and high quality of bioactive compounds. Additionally, CE0.5 is proposed as a potential source of natural antioxidants and anti-aging properties for further development of anti-wrinkle products

CHARACTERIZATION OF HYDRODISTILLATED POMELO PEEL OIL AND THE ENHANCEMENT OF BIOLOGICAL ACTIVITIES USING MICROEMULSION FORMULATIONS

Objective: The present study aims to investigate the compositions and biological activities of essential oil extracted from pomelo peel and develop into microemulsions.Methods: Four subspecies of pomelo including Kao-Namphung (KN), Kao-Puang (KP), Kao-Tang-Gwa (KT), and Kao-Yai (KY) were subjected to the hydrodistillation to yield essential oils. The constituents of each oil was analyzed by GC-MS. Radical scavenging activities were determined by ABTS and DPPH assays, whereas, lipid antioxidant activity was determined by linoleic acid peroxidation assay. Antityrosinase activity and safety on human PBMCs were also investigated. Pseudoternary phase diagrams were constructed to reveal the effects of each compostions on the microemulsion regions. The microemulsion was formulated and chracterized for the particle size, rheological behavior and biological activities.Results: Limonene was the major constituent in KN, KP, KT, and KY oil which was detected up to 86.19%, 85.76%, 79.36%, and 80.20%, respectively. Among four oils, KT oil exhibited the highest radical scavenging, antioxidant and antityrosinase activities. The MTT assay revealed that KT oil had no toxicity on human PBMCs. The microemulsion formulation (ME) containing 15% KT, 36% Tween 20, 9% PEG 400, and 40% water, were formulated and chracterized. ME was transparent liquid with the particle size of 90.28 ± 1.60 nm. ME exhibited the Newtonian flow behavior with low viscosity (16.78 ± 0.12 Pas). In a comparison with KT oil, ME show significant higher radical scavenging and antioxidant activities (p< 0.01).Conclusion: Development of microemulsion increased radical scavenging and antioxidant activities of KT oil and would be an attractive system for further development to effective topical products.Â

CURCUMIN-LOADED MULTI-VALENT LIGANDS CONJUGATED-NANOPARTICLES FOR ANTI-INFLAMMATORY ACTIVITY

Objective: The present study aims to develop curcumin-loaded muti-valent ligands conjugated-nanoparticlesfor targeted cell and study the biological activities including anti-inflammatory activity and kinetic cellular uptake.Methods: Curcumin encapsulated PLGA nanoparticles were formulated by solvent displacement method. The cIBR and cLABL peptides were conjugated on the surface of PLGA nanoparticles using carbodiimide reaction to produce curcumin encapsulated cIBR-cLABL-nanoparticles (cIBR-cLABL-NPs). The expression of LFA-1 and ICAM-1 on the membrane of U937 cells were determined by flow cytometry. Kinetic binding and internalization profile of cIBR-cLABL-NPs and untargeted nanoparticles were also investigated by flow cytometry. Safety profile on PBMC and cytotoxicity profile on U937 cells were evaluated using the MTT assay. Anti-inflammatory activity was determined using aprotein denaturation method.Results: The densities of cIBR and cLABL peptide on the surface of PLGA nanoparticles were 22.0±4.4 and 19.6±2.8 pmol/cm2, respectively. The particle size and total surface area of PLGA were 256.1 nm and 0.038 m2/g. The result showed that U937 cells expressed both LFA-1 and ICAM-1 proteins on their membranes indicating the possibility to use U937 cells as a targeted cell. According to the kinetic binding and internalization profiles, the cellular uptake of cIBR-cLABL-NPs was significantly higher than that of untargeted-NPs at all-time points indicating the specific uptake of cIBR-cLABL-NPs to target cell. Moreover, the rate of binding and internalization interpreted by the slope of linear regression of cIBR-cLABL-NPs was more rapid than that of untargeted-NPs. The MTT assay revealed the safety on human PBMC of cIBR-cLABL-NPs. The IC50 of free curcumin and curcumin-loaded cIBR-cLABL-NPs were 0.13 and 2.27 μg/ml, respectively. Protein denaturation assay presented the concentration dependenceinhibition of protein denaturation by curcumin and cIBR-cLABL-NPs indicating anti-inflammatory activities of free curcumin and encapsulated curcumin.Conclusion: cIBR-cLABL-nanoparicles increased the quantity of binding, rate of binding, and the internalization by target cells with safety profile on human PBMC. Moreover, the biological activity of encapsulating agent was maintained. Therefore, it could be used as a drug delivery system for encapsulating anti-infalmmatory agents.Â

Enhancement of acaricide activity of citronella oil after microemulsion preparation

The objectives of this study were to evaluate the preparation of microemulsions for citronella oil and subsequently to compare their acaricide efficacy as against conventional citronella oil. The chemical analysis of the commercial citronella oil used was carried out using an Agilent 6890 gas chromatograph (GC) coupled to an electron impact ionization (EI, 70 eV) with an HP 5973 mass selective detector (MSD). In comparison among the emulsifier systems, the mixture of Tween 20 with propylene glycol in ratios 3 : 1, 2 : 1, 1 : 1, 1 : 2, and 1 : 3, the pseudoternary phase diagrams were conducted and used to define the most suitable system for preparing citronella oil microemulsions and using them in acaricide efficacy testing. The citronella microemulsions in concentrations ranging from 0.39-25% w/w were prepared and tested for their acaricide efficacies using Adult Immersion Test and Larval Package Test. The acaricide efficacies were determined by percentages of mortalities of larval and adult ticks and the values of LC50 and LC99. Results show that the most suitable emulsifier system was in a ratio of 3 : 1. The physicochemical characteristics indicated that the size of the microemulsions were in the range of 19.6 ± 0.4 nm to 47.3 ± 2.3 nm with moderate polydispersity index (0.3-0.7). The microemulsions had stronger acaricide efficacy than the conventional citronella oil, indicated by significant lower LC50 and LC99 values. The LC99 of the microemulsions against larval and adult ticks at 24 h were 0.78% w/w (0.56-1.02) and 28.4% w/w (23.27-37.43), respectively. These results demonstrated that microemulsion preparation can be used to improve the acaricide efficacy of citronella oil

Enhancement of Cholinesterase Inhibition of Alpinia galanga (L.) Willd. Essential Oil by Microemulsions

This study aimed to investigate the chemical composition and reveal the selective inhibitory activity of Alpinia galanga (L.) Willd. essential oil (AGO) on acetylcholinesterase (AChE) compared to butyrylcholinesterase (BChE). The chemical composition of AGO was investigated by means of gas chromatography–mass spectrometry. Ellman’s method was used to determine the inhibitory activities against AChE and BChE. Microemulsion systems with desirable anticholinesterase effects were developed. Methyl cinnamate and 1,8-cineole were reported as the major component of AGO. The IC50 values of A. galanga oil against AChE and BChE were 24.6 ± 9.6 and 825.4 ± 340.1 µg/mL, respectively. The superior selectivity of AGO on AChE (34.8 ± 8.9) compared to galantamine hydrobromide (6.4 ± 1.5) suggested AGO to be an effective ingredient with fewer side effects for Alzheimer’s treatment. Interestingly, the microemulsion of AGO possessed significantly higher anticholinesterase activity than that of native oil alone. Therefore, microemulsion of AGO is a promising alternative approach for the treatment of Alzheimer’s disease