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.

Validated HPTLC and antioxidant activities for quality control of catechin in a fermented tea (Camellia sinensis var. assamica).

Miang, a Thai traditional fermented tea (Camellia sinensis var. assamica), is exploited as nutraceutical and cosmeceutical ingredients despite limited standardization studies. Thus, this research aimed to develop a simple and rapid method for miang quality control using catechin and high-performance thin-layer chromatography (HPTLC) validated according to the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) and the Association of Official Analytical Collaboration (AOAC). The developing solvent consisting of toluene: ethyl acetate: acetone: formic acid (6:6:6:1 v/v/v/v) showed acceptable specificity with R f value of 0.54 ± 0.02 and linearity with correlation coefficient of 0.9951. The recovery was 98.84%-103.53%, and the RSD of intra- and inter-day precision was 0.70%-3.00% and 1.93%-4.94%, respectively. Miang ethyl acetate fraction is suggested to be attractive ingredient due to rich catechin (25.78 ± 0.53%), prolonged stability at 40 ◦C, and strong antioxidants determined by the assays of ABTS (IC50 = 3.32 ± 0.74 mg/ml), FRAP (89.05 ± 15.49 mg equivalent of FeSO4/g), and inhibition of lipid peroxidation (IC50 = 4.36 ± 0.67 mg/ml).

FLT3, a prognostic biomarker for acute myeloid leukemia (AML): Quantitative monitoring with a simple anti-FLT3 interaction and flow cytometric method.

BACKGROUND: Overexpression of fms-like tyrosine kinase 3 (FLT3) protein in leukemia is highly related to poor prognosis and reduced survival rate in acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) patients. Simple but efficient quantification of FLT3 protein levels on the leukemic cell surface using flow cytometry had been developed for rapid determination of FLT3 on intact cell surface. METHODS: Quantitation protocol for FLT3 biomarker in clinical samples was developed and validated. Cell model selection for calibration curve construction was identified and evaluated. Selected antibody concentrations, cell density, and incubation time were evaluated for most appropriate conditions. Comparison of the developed FLT3 determination protocol with the conventional Western blot analysis was performed. RESULTS: EoL-1 cell line was selected for using as positive control cells. Calibration curve (20%-120% of FLT3 positive cells) and quality control (QC) levels were constructed and evaluated. The results demonstrated good linearity (r2  > 0.99). The intra- and inter-day precision and accuracy, expressed as the coefficient of variation (%CV) and % recovery, were <20% and fell in 80%-120% in all cases. When compared with Western blotting results, FLT3 protein expression levels in leukemia patient's bone marrow samples were demonstrated in the same trend. CONCLUSIONS: The effective, reliable, rapid, and economical analytical technique using the developed flow cytometric method was demonstrated for FLT3 protein determination on leukemic cell surface. This method provided a practical analysis of FLT-3 biomarker levels which is valuable for physician decision in acute leukemia treatment.

Enhancement of anti-inflammatory activity of polyphenolic flavonoid rutin by encapsulation.

The cellular mechanisms underlying the anti-inflammatory activity of rutin which has been found to have in vivo inhibitory effects merit more evaluation. The effects of rutin and encapsulated-rutin on lipopolysaccharide (LPS)-induced IL-6 secretion, NF-κB expression, as well as protein denaturation were investigated. The secretion of IL-6 was not found to have significantly reduced upon incubation with either rutin or encapsulated-rutin at all concentrations. At 100 µg/mL, the cells treated with encapsulated-rutin brought about slightly reduced IL-6 secretion but significantly inhibited NF-kB protein expression and protein denaturation in comparison with rutin. Inflammation can be resolved through many mechanisms. The inhibition of IL-6 and NF-kB can serve not only to terminate inflammation but also to inhibit other cytokines or mechanisms. Further investigations are necessary to clarify, verify and establish the anti-inflammatory mechanisms of rutin. Additionally, the encapsulation is an interesting technique for enhancing rutin activity.

Development of Microemulsion Delivery System of Essential Oil from Zingiber cassumunar Roxb. Rhizome for Improvement of Stability and Anti-Inflammatory Activity.

The present study aims to investigate the major constituents of the essential oil from Zingiber cassumunar rhizome (EO) and to develop microemulsions with enhanced chemical stability and anti-inflammatory activity of EO. The major constituents of EO were terpinen-4-ol (40.5 ± 6.6%) and sabinene (17.4 ± 1.4%) as determined by gas chromatography-mass spectrometry. These compounds were responsible for the anti-inflammatory activities of EO. Sabinene and terpinen-4-ol significantly reduced nuclear factor-kappa B (NF-kB) expression by 47 ± 5 and 78 ± 8%, respectively (p < 0.001) and significantly reduced the interleukin-6 (IL-6) secretion levels to 64 ± 4% (p < 0.05) and 50 ± 1% (p < 0.001), respectively. EO microemulsions, developed using the system of EO/Tween 20 and propylene glycol (2:1)/water, showed the internal droplet size in the range of 211.5 ± 63.3 to 366.7 ± 77.8 nm. Both EO and EO microemulsions were shown to be safe for human use since there was no apparent toxic effect on human peripheral blood mononuclear cells. Interestingly, EO microemulsion could significantly protect sabinene from the evaporation after heating-cooling stability test, which leads to a good stability and high efficacy. Moreover, EO microemulsions significantly enhanced the anti-inflammatory effect comparing to the native EO. Therefore, microemulsions were attractive delivery system for natural anti-inflammatory compounds since they could enhance both efficacy and stability of EO.

LFA-1 on leukemic cells as a target for therapy or drug delivery.

Leukemia therapeutics are aiming for improved efficacy by targeting molecular markers differentially expressed on cancerous cells. Lymphocyte function-associated antigen-1 (LFA-1) expression on various types of leukemia has been well studied. Here, the role and expression of LFA-1 on leukemic cells and the possibility of using this integrin as a target for drug delivery is reviewed. To support this rationale, experimental results were also included where cIBR, a cyclic peptide derived from a binding site of LFA-1, was conjugated to the surface of polymeric nanoparticles and used as a targeting ligand. These studies revealed a correlation of LFA-1 expression level on leukemic cell lines and binding and internalization of cIBR-NPs suggesting a differential binding and internalization of cIBR-NPs to leukemic cells overexpressing LFA-1. Nanoparticles conjugated with a cyclic peptide against an accessible molecular marker of disease hold promise as a selective drug delivery system for leukemia treatment.