Topical application of Zanthoxylum piperitum extract improves lateral canthal rhytides by inhibiting muscle contractions.

Facial wrinkles are the predominant phenotypes of skin aging. To date, one of the most effective ways to improve wrinkles is botulinum toxin type A (BoNT/A) injection, which inhibits muscle contractions by reducing acetylcholine release from neurons. However, since BoNT/A is a hazardous neurotoxin, the injection can only be performed by medical doctors and the procedure is only possible through invasive injection, causing inconveniences such as pain. To overcome these inconveniences, we tried to find a way to reduce wrinkles non-invasively via mechanisms similar to BoNT/A. We first designed in vitro assays to test BoNT/A-like muscle contraction inhibition in two different model systems. By using the assays, we identified Zanthoxylum piperitum (Z. piperitum) fruit extract as a BoNT-like reagent (27.7% decrease of muscle contraction rates by 1000 ppm of Z. piperitum extract treatment). Next, we determined mechanisms of how Z. piperitum extract decreases muscle contraction rates and found that the extract treatment inhibits electrical signal transduction in neurons. We also showed that among known components of Z. piperitum extract, quercitrin is responsible for muscle contraction inhibition. We further identified that Z. piperitum extract has synergistic effects with acetyl hexapeptide-8 and BoNT/A light chain, which are well-known BoNT-like peptides. Finally, we showed that topical treatment of the Z. piperitum extract indeed decreases facial wrinkles and treatment of Z. piperitum extract with acetyl hexapeptide-8 has a tendency to improve wrinkles synergistically (14.5% improvement on average). The synergistic effect of the combination is expected to improve wrinkles effectively by implementing the BoNT/A mechanisms in a non-invasive way.

Enhancement of exfoliating efficacy of L-carnitine with ion-pair method monitored by nuclear magnetic resonance spectroscopy.

Carnitine (CAR), an amino acid derivative, has great potential as a facial exfoliating agent owing to its calcium chelating property under weakly acidic or neutral conditions. However, its application is limited by its poor transdermal penetration. To optimise its exfoliation efficacy with minimal concentration, we propose the ion-pair method. The ionic interaction between CAR and a zwitterionic substance was successfully monitored by measuring conductivity. The alterations of penetration and exfoliation efficacy for CAR addition to different types of counter ions were investigated in vitro and in vivo. We found that hydrogenated soya phosphatidylcholine (HSC), an amphiphilic counter ion, significantly increases the stratum corneum penetration and exfoliation efficacy of CAR. The changes of the CAR-HSC ionic interaction in the presence of calcium ions were also investigated by 1H nuclear magnetic resonance (NMR) spectroscopy. The NMR spectra for amino groups of CAR first decreased with HSC and then gradually recovered and shifted as calcium ions were added. From the results, a noble exfoliating complex of CAR with high exfoliation efficacy could be proposed. Moreover, the results demonstrate that NMR spectroscopy is useful to obtain direct experimental evidence of the molecular dynamics simulations of the alteration of an exfoliating complex as it penetrates.

Amino acids disrupt calcium-dependent adhesion of stratum corneum.

In the stratum corneum, the intercellular junction made up of cadherin proteins provides the structural integrity of the framework. Ca2+ ions are known to play a key role in maintaining this junction. In this study, we hypothesized that Ca2+ chelation in stratum corneum will weaken the bond of the tissue and consequently promote exfoliation. Amino acids, ubiquitously existing as metabolites and building blocks of the body, have the molecular property to chelate Ca2+ ions. In the current study, we verified the Ca2+ chelating property of amino acids and demonstrated that amino acids can interfere with the interaction of cadherins, separate stratum corneum into pieces, and thereby stimulate the exfoliation process of skin. These results validate the importance of Ca2+ ion in the skin exfoliation process. Importantly, our findings indicate that amino acids may be efficiently used for improving skin conditions.

Identification and validation of amino acid-based mild exfoliating agents through a de novo screening method.

BACKGROUND: Due to internal or external factors, the desquamation (turnover) rate of the stratum corneum slows down, resulting in skin problems. Therefore, adjusting the exfoliation rate through cosmetics is an important issue. OBJECTIVE: This report aimed to develop exfoliating agents with lesser adverse effects and higher efficiency through an ex vivo screening method and in vivo turnover rate analysis of human skin. METHODS: Various molecules were evaluated by the ex vivo porcine skin exfoliation method and screened for their potential as effective agents. To confirm the effect and mechanism of each agent found, the exfoliation efficiency was evaluated. Each agent was also applied to the actual human skin to determine its efficacy and side effects. RESULTS: Despite the pH 6, carnitine and serine, which have similar or better efficiency compared to PHA, were selected. Based on the results, it was confirmed that calcium. And it is nonirritating to the human skin and increases the turnover rate (~130%). CONCLUSION: Amino acid-based exfoliating agents, such as carnitine and serine, were identified and verified to enhance the rate of exfoliation of the stratum corneum. It is expected that the improvement of dullness, mild acne, fine wrinkles, and pores through skin exfoliation in the field of cosmetics can be achieved safely through these agents.

Four-fold Channel-Nicked Human Ferritin Nanocages for Active Drug Loading and pH-Responsive Drug Release.

Human ferritins are emerging platforms for non-toxic protein-based drug delivery, owing to their intrinsic or acquirable targeting abilities to cancer cells and hollow cage structures for drug loading. However, reliable strategies for high-level drug encapsulation within ferritin cavities and prompt cellular drug release are still lacking. Ferritin nanocages were developed with partially opened hydrophobic channels, which provide stable routes for spontaneous and highly accumulated loading of FeII -conjugated drugs as well as pH-responsive rapid drug release at endoplasmic pH. Multiple cancer-related compounds, such as doxorubicin, curcumin, and quercetin, were actively and heavily loaded onto the prepared nicked ferritin. Drugs on these minimally modified ferritins were effectively delivered inside cancer cells with high toxicity.

A Rhizavidin Monomer with Nearly Multimeric Avidin-Like Binding Stability Against Biotin Conjugates.

Developing a monomeric form of an avidin-like protein with highly stable biotin binding properties has been a major challenge in biotin-avidin linking technology. Here we report a monomeric avidin-like protein-enhanced monoavidin-with off-rates almost comparable to those of multimeric avidin proteins against various biotin conjugates. Enhanced monoavidin (eMA) was developed from naturally dimeric rhizavidin by optimally maintaining protein rigidity during monomerization and additionally shielding the bound biotin by diverse engineering of the surface residues. eMA allowed the monovalent and nonperturbing labeling of head-group-biotinylated lipids in bilayer membranes. In addition, we fabricated an unprecedented 24-meric avidin probe by fusing eMA to a multimeric cage protein. The 24-meric avidin and eMA were utilized to demonstrate how artificial clustering of cell-surface proteins greatly enhances the internalization rates of assembled proteins on live cells.

A facile synthesis of fluorescent silver nanoclusters with human ferritin as a synthetic and interfacing ligand.

Water-soluble fluorescent silver nanoclusters (NCs) formed on biomolecule ligands have been extensively studied due to their great potential as new biocompatible fluorescent materials for biosensors. As synthetic ligands, proteins in particular can provide unique structures and functions to the assembled fluorescent silver clusters. A key challenge, however, is to develop appropriate protein ligands and synthetic approaches for cluster formation, especially using native aqueous solutions, to fully preserve the valuable properties of the protein templates. Here we report a human ferritin-templated synthesis of fluorescent silver NCs under neutral aqueous buffer conditions. The unique metal binding property of ferritin and an optimized silver ion reduction allowed us to produce highly stable fluorescent silver NCs that are steadily assembled in the cage-like ferritin proteins. The fluorescent clusters were also successfully assembled on genetically engineered ferritin with antibody-binding protein G. The resulting protein G-ferritin-silver NC complex fully retained the ferritin structure as well as the antibody binding ability. The present silver nanoclusters on ferritin (Ft-Ag NCs) also showed highly specific Cu(2+)-induced fluorescence quenching. By exploiting the large but stable nature of ferritin, we fabricated a highly robust and porous hydrogel sensor system for rapid Cu(2+) detection, where the Ft-Ag NCs were stably encapsulated in surface-bound hydrogels with large pore sizes. Our Ft-Ag NCs that are formed under native aqueous conditions will have great potential as a new fluorescent material with the high structural and functional diversities of ferritin.

Mesoporous silica nanoparticle-based cisplatin prodrug delivery and anticancer effect under reductive cellular environment.

This work demonstrates the judicious application of a prodrug delivery strategy to achieve a highly improved anticancer drug effect of cisplatin using mesoporous silica nanoparticles. Our effort primarily addressed several pressing needs to overcome various impediments such as toxicity concerns, rapid inactivation, and low drug efficiency of cisplatin prodrug. The developed delivery system utilizes fluorescent mesoporous silica nanoparticles as a template to host the cisplatin prodrug through a reducible linkage. The inactive oxidized Pt(iv) complex installed on the surface of the mesoporous silica nanoparticles in the prodrug conferred stability to cisplatin; however, in the reductive environment of cancer cell lines the active cisplatin form was regenerated. Prodrug-conjugated nanoparticles showed 63 times lower IC50 value than that of cisplatin in HeLa cell line. The delivery system not only demonstrates enhanced cellular uptake but also shows a high drug effect which should diminish the associated side effects. Furthermore, a new, easy and inexpensive fluorescent based Pt quantification method has been adopted instead of the commonly used ICP-based quantification method and this strategy of quantification could be elaborated to monitor fluorescent prodrug nanoparticles during real-time diagnosis.

Nano-engineered optimal templates for surface-plasmon enhanced Raman scattering.

We investigated the critical conditions to realize reliable and nano-engineered templates for surface-plasmon enhanced Raman scattering (SERS). Ultra-sensitive SERSs of thymine oligonucleotides were successfully realized on the template of Au nanoparticle arrays which were prepared by the combination of electron-beam lithography and post-chemical modification techniques. Drastic enhancement of Raman signal from the thymine oligonucleotides was only observed on the optimized templates, where the tuning of the plasmon resonance condition and the formation of the hot spots were both critical. Our results suggest that the artificial generation of reproducible and controlled hot spots can be achieved by our approach.

Solvent-free acid-catalyzed ring-opening of epoxidized oleochemicals using stearates/stearic acid, and its applications.

Toxic solvent and strong acid catalysts causing environmental issues have been mainly used for ring-opening of epoxidized oleochemicals. Here, we demonstrated that magnesium stearate (Mg-stearate) was a high efficient catalyst for solvent-free ring-opening of epoxidized methyl oleate, a model compound of midchain epoxide. Mg-stearate resulted in the highest yield (95%) and conversion rate (99%) toward midchain alkoxyesters under the same conditions (160 °C, 12 h) superior to other fatty acid derivatives such as a Lewis acid (lithium and sodium stearate) and Brønsted acid (stearic acid). Based on this chemical study, we synthesized biogrease and thermoplastic using epoxidized soybean oil (ESO) and Mg-stearate via one-pot, solvent-free, and purification-free process. Mg-stearate played a significant role as a reactant for epoxide ring-opening and as a thickener when excess loading rate was used; viscosity increased from 1800 to 4500 Pa·s at 25 °C when ESO:Mg-stearate increased from 1:1 equiv to 1:2, then behaved like thermoplastics (T(g) = -27 °C, T(m) = 90 °C) with 1:4.