Key Engineering Materials Vol. 819

Abstract: The total phenolic content and antioxidant activities of five edible mushrooms (Pleurotus djamor, Agaricus bisporus, Hericlum erinaceus, Coprinus atramentraris and P. ostreatus) were evaluated. The Coprinus atramentraris ethanolic extract (CE) showed the promising antioxidant property and had a strong relationship with phenolic compound content. Liposome-containing CE formulations, were prepared with various lipid composition. The liposome formulation, L1, which composed of phospholipids and cholesterol at ratio of 6:4 gave the highest entrapment efficiency (24.03%), small particle size (143.03 nm) and favorable particle size distribution (0.311) with small values of zeta potential (-30.2 mV). L1 showed good anti-elastase property (IC50 0.029) and no cytotoxicity effect to the human skin fibroblasts and melanoma cells even at the highest concentration of 1 mg/mL compared to the unformulated extract. The in vitro skin permeation studies, using human cadaver skin and modified Franz-diffusion cells, showed that the L1 was able to considerably increase the rate of permeation of phenolic compounds in L1 compared to the CE solution. These results reflect the use of L1 as active agents in cosmeceuticals.

Abstract: Modified coconut oil (MCO) has been reported as a promising material for eradication of various fungi which are the primary causes of tinea capitis. The antifungal shampoo is reported as an effective adjuvant therapy for fungi infection. In order to develop MCO as an active ingredient in antifungal shampoo, formulation factors affecting the physical and antifungal properties of shampoo including type and amount of surfactants were investigated. The results indicated that formula containing triethanolamine lauryl sulfate (TLE), ranging from 40-70% w/w, showed the most transparency and stable among the primary surfactants used, suggesting the good compatibility with MCO. An addition of polyoxyethylene 20 sorbitan monolaurate (PSL), an auxiliary surfactant, in the range from 5-40%w/w could enhance the clearness of shampoo while the suitable amount of PSL is also depended on TLE concentration. Various surfactant mixtures of TLE ranging from 40-60%w/w and 20-40% w/w of PSL were selected to formulate MCO shampoo. All formulations of shampoo showed high transparency, effective cleaning action, good stability and Newtonian flow behavior. Moreover, all formulations could inhibit T. rubrum and M. canis both before and after temperature cycling test. Therefore, this study revealed good feasibility of MCO shampoos for tinea capitis treatment.

Abstract: Andrographolide (AG) is a diterpenoid lactone found in Andrographis paniculata leaves and stems. It has excellent activity against various cancer cells, for example, skin cancer cells. However, application of AG for skin cancer treatment in clinical trials is limited due to its poor water solubility. To overcome this problem, oil in water AG-loaded nanoemulsion (AG-NE) would be prepared. The objectives of this study were to investigate physicochemical properties of AG-NE and to determine its activity against non-melanoma skin cancer cells. Nanoemulsion (NE) without AG (NE base) and NE containing AG (0.1%w/w) were prepared by high-pressure homogenization technique. They contained a mixture of Tween 80 and Span 80 (5:1) (10% w/w) as an emulsifier. Their droplet size, zeta potential and physical stability were evaluated. Cytotoxicity of AG and AG-NE to non-melanoma skin cancer cells (A-431 cells) and normal skin fibroblast cells (HFF-1 cells) were investigated. The results showed that NE base and AG-NE had droplet size in a nanometer range. They had low viscosity with the flow behavior consistent with Newtonian liquids. Although their zeta potential values were slightly low, they showed good physical stability against centrifugal force. AG and AG-NE were not toxic to HFF-1 cells, but they could induce apoptosis of A-431 cells with IC50 of 25.83 μg/ml and 58.32 mg/ml, respectively. Therefore, AG-NE has become possible to use for investigation of its efficacy and safety in animal models and clinical trials.

Abstract: Nanofibers have been widely used for tissue engineering. Using charged polymers for the preparation of nanofibers can be useful for the loading of substances or macromolecules. Dual charge nanofiber mats are expected to be able to immobilize both positively charged and negatively charged substances in one versatile nanofiber mat. The purpose of this study was to prepare and characterize dual-charge nanofibers generated from poly (vinyl alcohol) (PVA)/poly-(acrylic acid-co-maleic acid) (PAMA) and chitosan (CS)/PVA. The polymer solutions of PAMA/PVA (1:1.63 w/w) and CS/PVA (1:2.33 w/w) were electrospun to form the nanofibers using dual-jet electrospinning process. The obtained dual-charge nanofibers were thermally crosslinked by leaving the nanofibers in the oven at 110-130 °C for 0.5, 1, 3, 5 h. The appearance of the nanofiber mat was characterized by a scanning electron microscope (SEM), and the diameter of nanofibers were determined by an image analysis software (J-micro vision^®). The percentage water insolubilization and FT-IR spectra were also determined. The dual-size nanofiber mats with smooth and bead-free fibers were obtained. The diameter of the PAMA/PVA and CS/PVA fibers was 574.54 ± 142.98 nm and 225.69 ± 41.92 nm, respectively. The desirable temperature and time for the crosslink of the dual-charge nanofiber mats was 130 °C for 1 h which could provide a high insolubilization with water capacity of 93.22 ± 2.23%.

Abstract: Lipid-based formulations have been used as a widespread carrier to improve gene delivery. Niosomes, one type of lipid-based vesicular systems are produced from non-ionic surfactants which are generally inexpensive and potentially more stable than phospholipids. This article was to develop PEGylated cationic niosomes for DNA delivery. Thin film hydration and sonication method were applied for cationic niosomes. The niosome formulations were composed of Span 20, cholesterol (Chol) and plier-like cationic lipid B (PCL-B) with or without cholesterol-polyethylene glycol 2000 (Chol-PEG). The physicochemical properties of cationic niosomes and nioplexes were evaluated including particle size, zeta potential, DNA condensation and serum protection. The transfection efficiency and cell viability were examined in HeLa cells. The particle size and surface charge of PEGylated cationic niosome containing Span 20: Chol: PCL-B: Chol-PEG at the molar ratio of 2.5: 2.5: 1.5: 0.14 (N-PEG2) were 129.47 ± 2.15 nm and 25.93 ± 4.18 mV, respectively. These PEGylated cationic niosomes could condense pDNA into the nanosize particles and also enhance the serum protection ability for at least 6 h. Moreover, N-PEG2 exhibited high transfection efficiency in comparison with lipofectamine^® 2000 and low cytotoxicity. Therefore, the novel PEGylated cationic niosomes have the capability to develop as a promising potential carrier for DNA delivery.

Abstract: In this study, astaxanthin (ASTA), with potential anti-tyrosinase and anti-melanin synthesis in melanoma cells (B₁₆F₁₀) was developed as a cosmetic ingredient in the form of microemulsions (MEs). The results showed that ASTA (1 mg/mL) had no toxic effects on melanoma cells and it exhibited high potential for reduction of tyrosinase and melanin content, representing 80.57% and 75.86%, respectively. However, the use of ASTA is limited due to its low stability resulting from its decomposition under light, heat, and oxygen. In order to overcome this drawback, ASTA was encapsulated within ME. ASTA-MEs, consisting of 0.5% w/w of ASTA, oil, surfactant and water, were prepared using titration method.The effect of IPM concentration into microemulsions were investigated at 10 % w/w (ASTA-ME1) and 20% w/w (ASTA-ME2). The physical stability after accelerated condition of all the formulations was also investigated. The results indicated that a thermodynamically stable of microemulsion could improve the physical stability of ASTA. Nonetheless, the oil concentration had a slight influence on the physical stability of ASTA-ME1 and ASTA-ME2. In conclusion, nanoencapsulation can improve the physical stability of pigment extract to be used as a cosmetic ingredient in skin brightening products.

Abstract: This research aimed to develop clotrimazole (CT)-loaded mucoadhesive nanofiber patches for oral candidiasis. The three-layered sandwich-like nanofiber patches were prepared by electrospinning technique. The spinning solution for the middle layer composed of 8 %wt polyvinylpyrrolidone (PVP), 90 mM hydroxy propyl-β-cyclodextrin (HPβCD) and 10 % (wt to polymer) of CT in a solvent mixture of ethanol:water:benzyl alcohol. The outer layers were fabricated from a mixture of 1 %wt hyaluronic acid (HA) or catechol bearing hyaluronic acid (HA-cat) and 10 %wt polyvinyl alcohol (PVA) at varied weight ratios. The thickness of the outer layers was varied by adjusting the volume of coating polymer solution ranging from 1 to 3 mL. Desirable smooth nanosized fibers were obtained from the electrospinning process. Increasing the thickness of the outer layer brought about a significant increase in the fiber strength and flexibility. The viscosity of HA-cat/mucin mixture showed good polymer-mucin interaction indicating higher mucoadhesive property of the nanofibers. The drug loading capacity (LC) displayed the potential of the nanofibers for drug encapsulation. The highest LC value of 123.80 ± 5.61 μg/mg was obtained from the nanofibers coated with 1 mL of the coating solution. CT was rapidly released from the nanofiber in the first hour followed by a steady release. The released amount reach above 80% in 2 h. The nanofibers provided superior antifungal activity against Candida albicans compared to CT powder. Moreover, they were found to be nontoxic to the human gingival fibroblast cells. Thus, the sandwich nanofibers may be further developed to be a potential candidate for oral candidiasis treatment in the near future.

Abstract: Niosomes are a lipid nanoparticle which have been widely used as non-viral carrier for therapeutic DNA or siRNA. They are formulated from non-ionic surfactant and other helper lipids. The aim of this study were to formulate niosome containing spermine-based cationic lipid with different linkers and to evaluate the efficiency of siRNA delivery in cervical cancer cell (HeLa cell). The niosomes were formulated from cholesterol (Chol), Span 20 and different cationic lipid (Ay, By, Cy and Dy) at various molar ratios. The properties of niosomes and ability of niosome to complex with siRNA were characterized. The cellular uptake, gene silencing efficiency and cytotoxicity were also determined. From the results, niosomes formulated at Chol:Span20:lipid molar ratio of 2.5:2.5:2 showed positive zeta potential and they were in nanosize (<200 nm). The binding ability of cationic niosomes to siRNA depended on types of cationic lipid. Among niosome/siRNA complexes, the niosome By/siRNA complex provided the highest gene silencing efficiency at weight ratio of 20. The highest cellular uptake also obtained by using niosome By as a carrier. The cytotoxicity revealed that cationic niosomes had low toxicity (cell viability > 80%). In conclusion, the cationic niosomes prepared from Chol, Span 20 and spermine-based cationic lipids are able to complex with siRNA and suitable for siRNA delivery with low toxicity.

Abstract: Pueraria mirifica (PM) extract is locally used to promote hair growth. However, the effective transdermal delivery system should be prepared to deliver the extract through the skin barrier. The objective of this study was to develop solid lipid nanoparticles (SLN) containing PM ethanolic extract for hair growth promotion. The cell viability and proliferation of human follicle dermal papilla cells (HFDPCs) treated with PM extract were evaluated by MTT assay. SLN formulations were developed as a transdermal delivery system of the PM extract, compared with liposomes. The physicochemical properties of these nanoparticles were determined. The in vitro skin permeation study was also evaluated by Franz type diffusion cells. For the result, PM extract was a good safety herbal extract, which no cytotoxicity at the concentrations from 1 to 1,000 μg/ml. The cell proliferation of PM extract treated HFDPCs significantly increased in a dose-dependent manner, indicating the possibility to promote hair growth at the concentrations from 10 to 100 μg/ml. For formulation development, 5% (w/v) PM extract-loaded SLN exhibited small particle size (93.83 ± 0.32 nm) with narrow size distribution and negatively charged. This formulation had the highest percent entrapment efficiency (42.64 ± 0.47%), followed by SLN containing 1% (w/v) PM extract (8.84 ± 0.24%) and undetectable in liposomes. For the skin permeation result, SLN containing 5% (w/v) of PM extract could penetrate through the skin more than solution form. Therefore, the small particle size and high PM extract entrapped in SLN exhibited higher PM extract penetrated through the skin barrier and hair follicles than PM ethanolic extract solution. PM extract-loaded SLN might be an effective formulation for hair growth disorders treatment.

Abstract: Recently, coconut oil has become an attractive natural material consisting of monolaurin which exhibits antibacterial, antifungal, antiviral, and antiprotozoal effects. However, coconut oil is relatively immiscible with water, the main composition of human body. This study was thus focused on the development of nanoemulsions containing coconut oil employing the combination of several surfactants and variation of mixing speed for the purpose of overcoming the immiscible problem. Formulations of nanoemulsions were prepared within this study. The HLB values were then measured and used to determine the compatibility between coconut oil and surfactants. Moreover, the speed of homogenizer which might have a significant impact on the physical properties of nanoemulsions, was taken into account. Meanwhile, particle size, zeta potential, pH, long-term stability, and antibacterial activity were also examined. According to the results, the nanoemulsions prepared from 10% (w/w) of coconut oil and 10% (w/w) of surfactants comprising of Tween^® 80 and Span^® 80 in the ratio of 2 to 3 at the homogenization speed of 15000 rpm seemed to be suitable for topical administration with the characteristics as follows: the droplet size of 254.7±0.016 nm, the zeta potential of-4.41±1.46 mV, and the pH values of 6.13± 0.01. In addition, coconut oil formulated in the form of nanoemulsions demonstrated the efficient antimicrobial activities against Staphylococcus aureus, a gram-positive bacteria. In conclusion, this study represents the effect of mixing speed on the properties of nanoemulsions containing coconut oil which could be further developed as a substitute for topical antibiotics.