Lipid Monolayer on Cell Surface Protein Templates Functional Extracellular Lipid Assembly.

When the ancestors of men moved from aquatic habitats to the drylands, their evolutionary strategy to restrict water loss is to seal the skin surface with lipids. It is unknown how these rigid ceramide-dominated lipids with densely packed chains squeeze through narrow extracellular spaces and how they assemble into their complex multilamellar architecture. Here it is shown that the human corneocyte lipid envelope, a monolayer of ultralong covalently bound lipids on the cell surface protein, templates the functional barrier assembly by partly fluidizing and rearranging the free extracellular lipids in its vicinity during the sculpting of a functional skin lipid barrier. The lipid envelope also maintains the fluidity of the extracellular lipids during mechanical stress. This local lipid fluidization does not compromise the permeability barrier. The results provide new testable hypotheses about epidermal homeostasis and the pathophysiology underlying diseases with impaired lipid binding to corneocytes, such as congenital ichthyosis. In a broader sense, this lipoprotein-mediated fluidization of rigid (sphingo)lipid patches may also be relevant to lipid rafts and cellular signaling events and inspire new functional materials.

Homology Modeling, Molecular Docking, Molecular Dynamic Simulation, and Drug-Likeness of the Modified Alpha-Mangostin against the ß-Tubulin Protein of Acanthamoeba Keratitis.

Acanthamoeba species are capable of causing amoebic keratitis (AK). As a monotherapy, alpha-mangostin is effective for the treatment of AK; however, its bioavailability is quite poor. Moreover, the efficacy of therapy is contingent on the parasite and virulent strains. To improve readiness against AK, it is necessary to find other derivatives with accurate target identification. Beta-tubulin (BT) has been used as a target for anti-Acanthamoeba (A. keratitis). In this work, therefore, a model of the BT protein of A. keratitis was constructed by homology modeling utilizing the amino acid sequence from NCBI (GenBank: JQ417907.1). Ramachandran Plot was responsible for validating the protein PDB. The verified BT PDB was used for docking with the specified ligand. Based on an improved docking score compared to alpha-mangostin (AM), two modified compounds were identified: 1,6-dihydroxy-7-methoxy-2,8-bis(3-methylbut-2-en-1-yl)-9H-xanthen-9-one (C1) and 1,6-dihydroxy-2,8-bis(3-methylbut-2-en-1-yl)-9H-xanthen-9-one (C2). In addition, molecular dynamics simulations were conducted to analyze the interaction characteristics of the two bound BT-new compound complexes. During simulations, the TRP9, ARG50, VAL52, and GLN122 residues of BT-C1 that align to the identical residues in BT-AM generate consistent hydrogen bond interactions with 0-3 and 0-2. However, the BT-C2 complex has a different binding site, TYR 258, ILE 281, and SER 302, and can form more hydrogen bonds in the range 0-4. Therefore, this study reveals that C1 and C2 inhibit BT as an additive or synergistic effect; however, further in vitro and in vivo studies are needed.

In vitro galactose-targeted study of RSPP050-loaded micelles against liver hepatocellular carcinoma.

Andrographolide is a group of diterpenoid lactone isolated from Andrographis paniculata (Burm. F.) NEES. One of the analogues is 19-O-triphenylmethylandrographolide (RSPP050) which possesses anticancer activity. In seeking to capitalise on the last property, we have investigated the in vitro tumour targeting capabilities and MRI imaging for hepatocellular carcinoma. In this study, we have designed galactose-targeted and non-targeted micelles comprised of poly(ethylene glycol)-b-poly(lactide) that enveloped RSPP050 as an anticancer agent and superparamagnetic iron oxide (SPIO) as a contrast agent. The targeting abilities were endeavored by examining the cellular uptake with MTT assay, fluorescence microscopy, Prussian blue staining, and in vitro MRI. Targeted SPIO micelles as a T2* contrast agent decreased the relative T2* MRI intensity at 3 h. Results revealed that galactose micelles displayed 10.91 ± 0.19% drug loading content, -37.17 ± 0.63 mV zeta potential, and these micelles at the concentration of 0.5 µg/ml exhibited higher cytotoxicity than non-targeted micelles and free RSPP050 after incubation for 24 h. Fluorescence microscopy and Prussian blue staining at 3 h demonstrated significant cellular uptake by HepG2 cells. Thus, anticancer activity of RSPP050 could be improved using galactose as a targeting ligand and theranostic function was achieved using SPIO.

Sinigrin and Its Therapeutic Benefits.

Sinigrin (allyl-glucosinolate or 2-propenyl-glucosinolate) is a natural aliphatic glucosinolate present in plants of the Brassicaceae family, such as broccoli and brussels sprouts, and the seeds of Brassica nigra (mustard seeds) which contain high amounts of sinigrin. Since ancient times, mustard has been used by mankind for its culinary, as well as medicinal, properties. It has been systematically described and evaluated in the classical Ayurvedic texts. Studies conducted on the pharmacological activities of sinigrin have revealed anti-cancer, antibacterial, antifungal, antioxidant, anti-inflammatory, wound healing properties and biofumigation. This current review will bring concise information about the known therapeutic activities of sinigrin. However, the information on known biological activities is very limited and, hence, further studies still need to be conducted and its molecular mechanisms also need to be explored. This review on the therapeutic benefits of sinigrin can summarize current knowledge about this unique phytocompounds.

In vitro anti-cancer effects of artemisone nano-vesicular formulations on melanoma cells.

Artemisone is a 10-amino-artemisinin derivative that is markedly superior in vitro and in vivo to current artemisinins against malaria and also possesses antitumor activity. In seeking to capitalise on the last property, we have examined the encapsulation of artemisone in nano-vesicular niosomes and solid lipid nanoparticles, and have evaluated efficacies of the free and encapsulated artemisone against human melanoma A-375 cells and effects on human keratinocytes (HaCaT). Artemisone is successfully encapsulated into the nano-vesicles with encapsulation efficiencies of 67±6% and 79±5%, and with average particle sizes being 211±10nm and 295±18nm respectively. The formulations displayed highly selective cytotoxicity towards the melanoma cells with negligible toxicity towards the normal skin cells. The artemisone-loaded nano-vesicles almost completely inhibited the melanoma cells compared to the free drug. The results overall suggest a potentially more useful therapeutic strategy that needs to be evaluated for the treatment of melanoma and other cancers. FROM THE CLINICAL EDITOR: Apart from being an effective anti-malarial drug, a surprising action of artemisone also has antitumor activity. Nonetheless, its low water solubility and bioavailability has limited its clinical use. In this article, the authors enacapsulated artemisone in nano- vesicles and solid lipid nano-particles (SLNs). In-vitro studies confirmed the selective cytotoxicity towards melanoma cells. Further in-vivo and pre-clinical studies are awaited.

Alginate/pectin dressing with niosomal mangosteen extract for enhanced wound healing: evaluating skin irritation by structure-activity relationship.

Most modern wound dressings assist the wound-healing process. In contrast, conventional wound dressings have limited antibacterial activity and promote sporadic fibroblast growth. Therefore, wound dressings with prolonged substance release must be improved. This research aimed to develop hydrogel films. These were synthesized from alginate and pectin, incorporated with mangosteen extract (ME), and encapsulated in niosomes (ME-loaded niosomes). Subsequently, we examined the in vitro release and physical characteristics of ME-loaded niosomes. These characteristics included particle pH, size, charge, polydispersity index (PDI), and drug loading properties. These properties included drug loading content (DLC), entrapment efficiency (EE), and yield (Y). Additionally, we examined the swelling ratio and biological characteristics of the hydrogel film. These characteristics included antibacterial activity, cytotoxicity (L929), cell attachment to the tested materials, cell migration, hemocompatibility, and in vivo irritation. Significant results were obtained using a 2:1 niosome preparation containing Span60 and cholesterol. Ratio influenced size, charge, PDI, DLC, EE, and Y. The results were 225.5 ± 5.83 nm, negatively charged, 0.38, 16.2 ± 0.87%, 64.8 ± 3.49%, and 87.3 ± 3.09%, respectively. Additionally, the release of encapsulated ME was pH sensitive because 85% of the ME can be released at a pH of 5.5 within seven days and decrease to 70% at a pH of 7.4. The maximum swelling ratios of patches with 0.5% and 1% Ca2+ crosslinking were 867 wt% and 1,025 wt%, respectively, after 30 min. These results suggested that a medium dose (15 mg) of niosomal ME incorporated in a hydrogel film provided better bacterial inhibition, cell migration, and cell adhesion in an in vitro model. Additionally, no toxicity was observed in the fibroblasts and red blood cells. Therefore, given the above-mentioned advantages, this product can be a promising candidate for wound dressing applications.

Acidic pH Is Required for the Multilamellar Assembly of Skin Barrier Lipids In Vitro.

Lipid membrane remodeling belongs to the most fundamental processes in the body. The skin barrier lipids, which are ceramide dominant and highly rigid, must attain an unusual multilamellar nanostructure with long periodicity to restrict water loss and prevent the entry of potentially harmful environmental factors. Our data suggest that the skin acid mantle, apart from regulating enzyme activities and keeping away pathogens, may also be a prerequisite for the multilamellar assembly of the skin barrier lipids. Atomic force microscopy on monolayers composed of synthetic or human stratum corneum lipids showed multilayer formation (approximately 10-nm step height) in an acidic but not in a neutral environment. X-ray diffraction, Fourier transform infrared spectroscopy, and permeability studies showed markedly altered lipid nanostructure and increased water loss at neutral pH compared with that at acidic pH. These findings are consistent with the data on the altered organization of skin lipids and increased transepidermal water loss under conditions such as inadequate skin acidification, for example, in neonates, the elderly, and patients with atopic dermatitis.

Layer-by-layer nanocoating of antibacterial niosome on orthopedic implant.

The major clinical hindrance of orthopedic implants is the bacterial infection, which can lead to biofilm formation and ultimately results in implant rejection. In this research, layer-by-layer nanocoating consists of vancomycin/PLA/vancomycin-loaded niosomes was designed. Vancomycin-loaded niosomes were formulated by thin film hydration method and the attributes of niosomes in terms of size, zeta potential, drug loading and EE, were assessed. The size was 340.5 ±â€¯2.95 nm with the zeta potential and %EE was 45.4 ±â€¯0.77 mV and 50.47 ±â€¯3.66% respectively. The dip coating technique was used to deposit a thin film, which was characterized morphologically under FE-SEM. Drug release from coated bone plates with and without vancomycin-loaded niosomes was also studied and results suggested that bone plates coated with vancomycin-loaded niosomes have accumulated more vancomycin than the control group and hence aided in the prolonged release up to two weeks. These niosomes-coated bone plates demonstrated superior antibacterial activity for longer time period, without exhibiting any cytotoxic effects towards normal cells (L929). These findings offer a promising approach to control the bacterial colonization and biofilms formation. This thin film nano-coating can also be utilized in coating of other medical devices, which are prone to infections.

In vitro wound healing and cytotoxic activity of the gel and whole-leaf materials from selected aloe species.

ETHNOPHARMACOLOGICAL RELEVANCE: Aloe vera is one of the most important medicinal plants in the world with applications in the cosmetic industry and also in the tonic or health drink product market. Different parts of Aloe ferox and Aloe marlothii are used as traditional medicines for different applications. Although wound healing has been shown for certain aloe gel materials (e.g. A. vera ) previously, there are conflicting reports on this medicinal application of aloe leaf gel materials. AIM OF THE STUDY: The present study aimed at determining the wound healing properties of the gel and whole-leaf materials of Aloe vera, Aloe ferox and Aloe marlothii, as well as their cytotoxic effects on normal human keratinocyte cells (HaCaT). MATERIALS AND METHODS: Nuclear magnetic resonance spectroscopy was used to chemically fingerprint the aloe gel and whole-leaf materials by identifying characteristic marker molecules of aloe gel and whole-leaf materials. An MTT assay was performed to determine the cytotoxicity of the various aloe whole-leaf and gel materials on HaCaT cells. Wound healing and in vitro cell migration were investigated with HaCaT cells by means of the CytoSelect™ assay kit. RESULTS: The in vitro wound healing assay suggested that all the aloe gel and whole-leaf materials examined, exhibited faster wound healing activity than the untreated control group. After 48h, all the aloe gel and whole-leaf materials almost completely caused full wound closure, displaying 98.07% (A. marlothii whole-leaf), 98.00% (A. vera gel), 97.20% (A. marlothii gel), 96.00% (A. vera whole-leaf), 94.00% (A. ferox gel) and 81.30% (A. ferox whole-leaf) wound closure, respectively. It was noteworthy that the gel materials of all the three aloe species exhibited significantly faster (p<0.05) wound healing actions when compared to their respective whole-leaf materials at 32h. CONCLUSION: The gel and whole-leaf materials of A. vera, A. ferox and A. marlothii have shown the ability to heal wounds at a faster rate and to a larger extent than untreated keratinocytes. The MTT assay results suggested that the gel and whole-leaf materials of all the selected Aloe species showed negligible toxicity towards the HaCaT cells.

In vitro wound healing and cytotoxic effects of sinigrin-phytosome complex.

Sinigrin is a class of glucosinolates found naturally in plants of the Brassicaceae family. Lately, studies have shown that sinigrin possesses anticancer, antibacterial and anti-inflammatory activities. Since its efficacy has not been explored on wound healing, we examined the effect of sinigrin on HaCaT cells. We also aimed at formulating sinigrin into phytosome to form a sinigrin-phytosome complex and study the wound healing and cytotoxic activities on A-375 and HaCaT cells. Sinigrin was efficiently formulated into the phytosome with an average particle size of 153 ± 39 nm, zeta potential of 10.09 ± 0.98 mV and complex efficiency of 69.5 ± 5%. The formation of the sinigrin-phytosome complex was confirmed by DSC and FTIR analysis. The sinigrin-phytosome complex significantly exhibited wound healing effects when compared to sinigrin alone. After 42 h, the phytosome complex completely healed the wound, whereas sinigrin alone showed only 71% wound closure. The sinigrin-phytosome complex displayed minimal toxicity towards HaCaT cells and at higher concentrations, it showed potent activity towards A-375. The results indicated that sinigrin-phytosome complex augmented the therapeutic potential of sinigrin towards the wound healing activity and this approach should be explored further for cancerous wound treatment.

In vitro skin permeation of sinigrin from its phytosome complex.

OBJECTIVES: Sinigrin is a major glucosinolate present in plants of the Brassicaceae family. Recently, sinigrin and its phytosome formulations have been investigated for its wound-healing actions, by our research group. The aim of this study was to demonstrate sinigrin drug release from its phytosome complex and also to determine whether the phytosome complex enhances the delivery of sinigrin into the skin when compared to free sinigrin. METHODS: In vitro Franz cell diffusion studies were performed on human abdominal skin. The morphology of the phytosome complex was examined by transmission electron microscopy. The in vitro drug release was determined using dialysis sacks. KEY FINDINGS: The in vitro drug release indicated a controlled and sustained release of sinigrin from the phytosome complex. Tape stripping results showed that the sinigrin-phytosome complex (0.5155 µg/ml) statistically significantly enhanced the delivery of sinigrin into the stratum corneum-epidermis when compared to the free sinigrin (0.0730 µg/ml). CONCLUSIONS: These results suggested the possibility of utilizing sinigrin-phytosome complex, to optimally deliver sinigrin to the skin which can be further used for various skin-related diseases including wound healing.

In vitro skin permeation of artemisone and its nano-vesicular formulations.

The artemisinin derivative artemisone has antitumor activity. In particular when encapsulated in solid lipid nanoparticles (SLNs) and niosomes, it is active against human melanoma A-375 cells, although such formulations have a negligible effect on human keratinocyte cells. The aim here was to determine whether these formulations could enhance the topical delivery and skin permeation of artemisone as a prelude to evaluating use of artemisone and related compounds for melanoma treatment. In vitro skin permeation studies were conducted to determine the concentration of artemisone delivered into the stratum corneum-epidermis and epidermis-dermis. Artemisone-SLNs delivered artemisone into the stratum corneum-epidermis at significantly higher concentration (62.632 µg/mL) than the artemisone-niosomes (12.792 µg/mL). Neither of the controls delivered artemisone into the stratum corneum-epidermis. In the epidermis-dermis, artemisone (13.404 µg/mL) was only detected after application of the SLN formulation. Overall, the excellent topical delivery of artemisone with the SLN formulation coupled with the intrinsic activity of formulated artemisone confirms potential for use in treatment of melanoma.