BALB/c mice (male, 6−8 weeks old) were obtained from Orient Bio Co., Ltd. (Gyeonggi-do, Republic of Korea). The mice were housed in animal facility with environmental conditions (temperature of 22 ± 2°C, a humidity of 60 ± 10%, and a 12-h light/dark cycle), with free access to food and water. This experimental protocol was approved by the Committee for Animal Welfare at Daejeon University (DJUARB2019-021) and was performed in accordance with the committee guidelines. The mice received intranasal administration of a mixture of PM10 (ERM CZ-120, Sigma-Aldrich, USA) and diesel exhaust particles (DEP, SRM 2975, Sigma-Aldrich, St. Louis, MO, USA) on days 4, 7, and 10 and were orally treated with L. paracasei ATG-E1 every other day for 12 days. PM10 (3 mg/mL) and DEP (0.6 mg/mL) were dissolved in 1% aluminum hydroxide gel adjuvant. The mice were divided into following groups (n = 8 per group): (1) 1% aluminum hydroxide gel adjuvant-treated mice (NC), (2) PM10 + DEP-sensitized mice (PM10D-CTL), (3) 4 × 1010 CFU of ATG-E1-treated PM10 + DEP-sensitized mice (PM10D-ATG-E1), and (4) 3 mg/kg dexamethasone-treated PM10 + DEP-sensitized mice (PM10D-Dexa, positive control). All mice were sacrificed and blood, bronchoalveolar lavage fluid (BALF), and lung and intestinal tissues were harvested on day 12.
Erm cz120
Manufactured by Merck Group
Sourced in United States
The ERM-CZ120 is a laboratory-grade centrifuge designed for general-purpose use. It is capable of reaching speeds up to 12,000 RPM with a maximum RCF of 20,800 x g. The centrifuge features a versatile rotor accommodating various sample tube sizes and volumes. Key specifications include a brushless motor, digital speed and time controls, and automatic rotor identification.
Automatically generated - may contain errors
Lab products found in correlation
Srm 2975, by Merck Group (1 mentions)
Balb c mice, by Orient Bio (1 mentions)
M16 medium, by Merck Group (1 mentions)
Epigallocatechin gallate (egcg), by Merck Group (1 mentions)
Resveratrol, by Merck Group (1 mentions)
Punicalagin, by Merck Group (1 mentions)
Fine dust pm10 like, by Merck Group (1 mentions)
Ellagic acid, by Merck Group (1 mentions)
Erm cz100, by Merck Group (1 mentions)
8 protocols using erm cz120
1
Intranasal PM10 and DEP-induced Allergy Study
2
Particulate Matter Exposure in Mice
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PM was administered to the YPM, APM, YPMEX, and APMEX groups three times a week for 8 weeks. The PM used in the experiment was Fine dust (PM10-like, ERM-CZ120), which was purchased from Sigma-Aldrich (St. Louis, MO, USA), and whose PM10-like composition and concentration are certified by the European Reference Materials (ERM). Following the methodology described by Bai and van Eeden [28 (link)], 15 μg of PM was suspended in 200 μL of saline and 0.5 μg of PM per weight (in g) of mice was injected into the relevant groups via the tail vein. For the YCO, ACO, YEX, and AEX groups, 200 μL of saline was intravenously administered via the tail vein.
3
In vitro PM10 exposure protocol
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For the in vitro study, PM10 (PM10-like, European reference material ERM-CZ120) (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in phosphate buffered saline (PBS) at a concentration of 5 mg/ml and diluted to a range of concentrations with the basal medium of each cell type. The PM10 suspension was then sonicated for 30 minutes to prevent aggregation. Experiments were conducted within 1 hour of PM preparation to avoid variability in PM components at different concentrations between replicates.
4
PM10 Exposure in Mouse Oocyte
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All animal studies were approved and conducted according to the guidelines of the Animal Research Committee of Korea Research Institute of Bioscience and Biotechnology (KRIBB-AEC-19126). Germinal vesicle (GV) intact oocytes were collected from the ovaries of 6–8-week-old CD-1 mice and cultured in M16 medium (Sigma-Aldrich, St. Louis, MO) with an overlaying mineral oil layer at 37°C under 5% CO2.
For PM10 (#ERM-CZ120; Sigma Aldrich) treatment, oocytes were cultured in media supplemented with concentrated PM10. Before the experiment, PM10 was dissolved in M16 medium according to the experimental concentration (1, 5, and 10 mg/mL), and pre-warmed at 37°C at least overnight. Thereafter, only completely dissolved PM10 was obtained by filtration through a 0.22-μm filter. Oocytes were then cultured in PM, which was 100% soluble.
For PM10 (#ERM-CZ120; Sigma Aldrich) treatment, oocytes were cultured in media supplemented with concentrated PM10. Before the experiment, PM10 was dissolved in M16 medium according to the experimental concentration (1, 5, and 10 mg/mL), and pre-warmed at 37°C at least overnight. Thereafter, only completely dissolved PM10 was obtained by filtration through a 0.22-μm filter. Oocytes were then cultured in PM, which was 100% soluble.
5
Acne-Inducing C. acnes Mouse Model
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Female 6-week-old HR-1 mice were purchased from SLC (Japan) and acclimatized for 1 week. C. acnes was obtained and isolated from the pustules of Korean patients with acne. This study was conducted using our previously reported inflammatory acne mouse model.16 (link)
C. acnes were injected at 109 colony-forming units/20 μL into four sites on a mouse’s back using a 30-gauge needle. After 1 week, 100 μL of 100 μg/mL PM10 (PM10-like, European reference material ERM-CZ120; Sigma-Aldrich, St Louis, MO, USA) and 5 μM punicalagin, 1 μM EGCG, or 1 μM resveratrol (Sigma-Aldrich) were applied to the treated back skin for 2 weeks. All mice (3 for punicalagin, 3 for EGCG, and 3 for resveratrol) were sacrificed by CO2 inhalation to obtain dorsal skin samples. Tissue samples, including the inflammatory nodules, were cut to 1 × 1 cm for RNA isolation and tissue staining. The experiment was conducted to contain the same amount of RNA after RNA isolation. The experimental scheme is presented inSupplemental material Figure 2 . Animal experiments were conducted from 2020 at the animal experiment center managed by the School of Medicine, Kyungpook National University. Our study design was approved by the Institutional Animal Care and Use Committee of Kyungpook National University (IRB Number KNU 2021-0068) and followed the Guide for the Care and Use of Laboratory Animals.
C. acnes were injected at 109 colony-forming units/20 μL into four sites on a mouse’s back using a 30-gauge needle. After 1 week, 100 μL of 100 μg/mL PM10 (PM10-like, European reference material ERM-CZ120; Sigma-Aldrich, St Louis, MO, USA) and 5 μM punicalagin, 1 μM EGCG, or 1 μM resveratrol (Sigma-Aldrich) were applied to the treated back skin for 2 weeks. All mice (3 for punicalagin, 3 for EGCG, and 3 for resveratrol) were sacrificed by CO2 inhalation to obtain dorsal skin samples. Tissue samples, including the inflammatory nodules, were cut to 1 × 1 cm for RNA isolation and tissue staining. The experiment was conducted to contain the same amount of RNA after RNA isolation. The experimental scheme is presented in
6
Punicalagin and Ellagic Acid Extraction
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Punicalagin (purity > 98%, a mixture of 40% α and 60% β anomers) and ellagic acid (purity > 98%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fine dust (PM10-like) (European reference material ERM-CZ120) was purchased from Sigma-Aldrich. PPE was obtained from Hwasoomok Co. (Youngchen, Korea). The extract was prepared by extracting dry raw materials with water at 55°C for 2 h, followed by concentration and spray drying.
7
Preparation of Standard PM Particulates
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PM2.5, which is a standard diesel PM (SRM1650b) issued by the National Institute of Standard and Technology (Gaithersburg, MD, USA), was bought from Sigma-Aldrich (St. Louis, MO, USA). It was dissolved in dimethyl sulfoxide (DMSO) at 50-mg/ml concentration. PM10-like fine dusts (ERM-CZ100 and ERM-CZ120), which are issued by the European Reference Materials (ERM, Belgium), were brought from Sigma-Aldrich. The former (PM10-PAH) includes several PAHs (benzoanthracene, benzopyrene, benzofluoranthene, and dibenzoanthracene, etc.) in ambient PM10, the latter (PM10) contains heavy metals (arsenic, cadmium, lead, and nickel). They were suspended in phosphate buffered saline (PBS) at 5-mg/ml concentration. PM was prepared just before cell application and sonicated in an ultrasonic bath for 10 min to avoid variability in PM composition and aggregation of particles.
8
Dieckol Alleviates Acne-Induced Inflammation
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Six-week-old female HR-1 mice were purchased from SLC Inc. (Hamamatsu, Japan) and acclimatized for 1 week. We obtained and isolated C. acnes from the pustules of the Korean patients with acne. We injected C. acnes as 109 colony-forming units/20 µl at four sites on the backs of the mice using a 30-gage needle. One week after injection, we applied the back skin of the mice with 100 µl of 100-µg/ml PM10 (PM10-like European reference material ERM-CZ120; Sigma, Burlington, MA, USA) or 5 µM of dieckol for 2 weeks. In addition, mice were also applied separately with 100 µl of 100-µg/ml PM10 in the morning and with 5 µM of dieckol in the afternoon to observe changes in the PM-induced gene expression by Dieckol. We measured the size of the C. acnes suspension-induced inflammatory nodules at weeks 1 and 2 after treatment.
We followed the experiment guidelines for the use of laboratory animals. The study was permitted by the Institutional Animal Care and Use Committee of Kyungpook National University (IACUC no. KNU 2021-0068).
We followed the experiment guidelines for the use of laboratory animals. The study was permitted by the Institutional Animal Care and Use Committee of Kyungpook National University (IACUC no. KNU 2021-0068).
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