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J Microbiol Biotechnol
2022 Nov 28;3211:1382-1389. doi: 10.4014/jmb.2207.07018.
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Asterias pectinifera-Derived Collagen Peptides Mixed with Halocynthia roretzi Extracts Exhibit Anti-Photoaging Activities during Exposure to UV Irradiation, and Antibacterial Properties.
Oh SJ
,
Park JY
,
Won B
,
Oh YT
,
Yang SC
,
Shin OS
.
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Asterias pectinifera, a species of starfish and cause of concern in the aquaculture industry, was recently identified as a source of non-toxic and highly water-soluble collagen peptides. In this study, we investigated the antioxidant and anti-photoaging functions of compounds formulated using collagen peptides from extracts of Asterias pectinifera and Halocynthia roretzi (AH). Our results showed that AH compounds have various skin protective functions, including antioxidant effects, determined by measuring the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl radicals, as well as anti-melanogenic effects, determined by measuring tyrosinase inhibition activity. To determine whether ethosome-encapsulated AH compounds (E(AH)) exert ultraviolet (UV)-protective effects, human dermal fibroblasts or keratinocytes were incubated with E(AH) before and after exposure to UVA or UVB. E(AH) treatment led to inhibition of photoaging-induced secretion of matrix metalloproteinase-1 and interleukin-6 and -8, which are associated with inflammatory responses during UV irradiation. Finally, the antibacterial effects of AH and E(AH) were confirmed against both gram-negative and gram-positive bacteria. Our results indicate that E(AH) has the potential for use in the development of cosmetics with a range of skin protective functions.
Fig. 1. Asterias pectinifera-derived collagen peptides mixed with Halocynthia roretzi extracts (AH) show antioxidant activity and tyrosinase inhibitory effect.(A) To evaluate the extracellular antioxidant level of the AH extracts, the scavenging activities of DPPH radicals were measured with various concentrations of AH extracts. Ascorbic acid (AA) was used as a positive control (1 mg/ml). (B) Various concentrations of AH were incubated with the tyrosine and tyrosinase solution and a mushroom tyrosinase inhibition (MTI) assay was performed. AA was used as a positive control (0.3 mg/ml). Values are expressed as the mean ± SD of three determinations. Statistical analysis: ***p < 0.001 vs. control (Ctl); ##p < 0.01, ###p < 0.001 vs. each concentration of AH01.
Fig. 2. Ethosome-encapsulated AH (E(AH)) shows antioxidant activity and tyrosinase inhibitory effect.(A) The scavenging activities of DPPH radicals by E(AH) at various concentrations were measured. Ascorbic acid (AA) was used as a positive control (0.03 mg/ml). (B) Mushroom tyrosinase inhibition (MTI) assay was performed using E(AH) compounds. Statistical analysis: ***p < 0.001 vs. Ctl.
Fig. 3. E(AH) treatment does not cause cytotoxicity in UV-irradiated cultured human epidermal keratinocytes (HaCaT) and human dermal fibroblasts (HDF).(A) A schematic view of the MTT assay in E(AH)-treated HaCaT and HDF cells. (B) HaCaT and HDF cells were treated with various concentrations of E(AH). After incubation, MTT assay was performed to measure the cell viability of E(AH). All data were indicated as mean ± SD of at least three independent experiments. Statistical analysis: *p < 0.05, **p < 0.01. (C) Schematic views of the MTT assay in UV-irradiated cells before and after treatment of the E(AH). (D) HaCaT and HDF cells were treated with either a control (Ctl), epigallocatechin gallate (EGCG), or various concentrations of E(AH) before or after irradiation with UVA (3.0 J/cm2) or UVB (0.03 J/cm2). At 48 h, cell viability was examined by MTT assay. EGCG was used as a positive control for the following experiments. All data were indicated as mean ± SD of at least three independent experiments. Statistical analysis: *p < 0.05, **p < 0.01, ***p < 0.001 vs. Ctl-treated cells.
Fig. 4. E(AH) treatment in human epidermal keratinocytes (HaCaT) inhibits the secretion of photoaginginduced matrix metalloproteinase-1 (MMP-1) and cytokines.HaCaT cells were treated with a Ctl, EGCG, or various concentrations of E(AH) before or after irradiation with UVB (0.03 J/cm2). After 48 h, (A) MMP-1, (B) IL-6, and (C) IL-8 secretion levels in the supernatant were measured by ELISA assay. All data were indicated as mean ± SD of at least three independent experiments. Statistical analysis: *p < 0.05, **p < 0.01, ***p < 0.001 vs. Ctl-treated cells.
Fig. 5. E(AH) treatment in human dermal fibroblasts (HDF) leads to inhibition of photoaging-induced secretion of MMP-1 and cytokines.HDF cells were treated with a Ctl, EGCG, or various concentrations of E(AH) before or after irradiation with UVA (3.0 J/cm2). After 48 h, (A) MMP-1, (B) IL-6, and (C) IL-8 secretion levels in the supernatant were measured by ELISA assay. All data were indicated as mean ± SD of at least three independent experiments. Statistical analysis: *p < 0.05, **p < 0.01, ***p < 0.001 vs. Ctl-treated cells.
Fig. 6. AH and E(AH) show antibacterial activities against multiple bacterial strains.(A) For paper disc diffusion assay S. aureus, E. coli, and P. aeruginosa were cultured overnight. Filter paper discs (8 mm) were soaked in Halocynthia roretzi extracts and placed on the plate and incubated at 37°C for 24 h. Then, the diameter of the clear zone was measured. (B) A paper disc diffusion assay was performed to examine the antibacterial effect of AH compounds against S. aureus, E. coli, and P. aeruginosa. (C) Bacteria in the exponential growth phase were inoculated in Halocynthia roretzi extracts or AH compoundcontaining media broth. After overnight incubation, absorbance at 600 nm was measured. (D) AH05 compound or E(AH) were added to bacterial suspensions of S. aureus, E. coli, and P. aeruginosa. Next day, bacterial suspensions were serially diluted and plated onto the LB or TSB plate to measure the colony-forming units. All data were indicated as mean ± SD of at least three independent experiments.
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