C57BL/6J mice were purchased (CLEA Japan INC, Tokyo, Japan) and were maintained in the animal facility at the Jikei University School of Medicine. All experimental procedures are approved by the Jikei University School of Medicine Animal Care Committee (#25031). A dose of 3 U/kg bleomycin (Nippon Kayaku Co., Tokyo, Japan) was intratracheally administered in 50 μL saline using MicroSprayer™ Aerosolizer and a high pressure syringe (PennCentury, Philadelphia, PA). Intraperitoneal dose of metformin (300 mg/kg) were given from day 7 to day 20. On the 21th day the lungs were removed. The lungs were fixed overnight in 10 % buffered formalin, embedded in paraffin, and the sections are stained with hematoxylin & eosin (HE).
High pressure syringe
Manufactured by Penn-Century
Sourced in United States
The High Pressure Syringe is a precision instrument designed to accurately deliver liquids at high pressures. It features a durable construction and a precise mechanism to control the amount and rate of liquid flow. The core function of this syringe is to provide a reliable and consistent way to apply pressurized liquid for various applications.
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3 protocols using high pressure syringe
1
Bleomycin-Induced Pulmonary Fibrosis in Mice
2
BALB/c Mice Lung Inflammation Model
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The 6−8-week old BALB/c mice were purchased from Jinan Pengyue Experimental Animal Breeding Co. LTD (Certificate No. SCXK 20190003, Shandong, China). The animals were fed with standard commercial diets and water and housed in independent ventilated cages in a temperature-controlled animal house (25 ± 2 °C) with a 12-h day/night cycle. The animals were allowed to acclimatize for 1 week. The experiment was carried out according to the Declaration of Helsinki, and all animal protocols and procedures were approved by the Animal Care and Use Committee of Shandong University (No. 2016020, Jinan, Shandong, China). The mice were randomly divided into 5 groups (n = 5): control group (C), model group (M), LPS + SAMC 20 group (20 mg/kg, L), LPS + SAMC 40 group (40 mg/kg, H), LPS + NAC group (500 mg/kg, P). After anesthesia, LPS (10 mg/kg) was instilled into the lungs of mice through a high-pressure syringe (Penn-Century, PA, USA). Then, mice were orally administrated with SAMC, NAC or the vehicle after 6 and 18 h, respectively. Control mice received the same volume of saline. All mice were sacrificed at 24 h.
3
Ameliorating Bleomycin-Induced Lung Fibrosis
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Animal experiments were performed in compliance with the guidelines of the Institute for Laboratory Animal Research, The Jikei University School of Medicine (Number: 2018–071). C57BL/6J mice (Charles River Laboratories) were used in all experiments. A dose of 2.5 U/kg bleomycin (Nippon Kayaku Co., 4234400D4032) was intratracheally administered in 50 ml saline using the MicroSprayer Aerosolizer and a high‐pressure syringe (PennCentury, Philadelphia, PA, USA). HBEC EVs, BM‐MSC EVs and NHDF EVs (2.0×109/body) were given intratracheally on day 7 and day 14 after bleomycin administration. Equivalent volumes of 0.9% normal saline were used as controls. On the 17th day, the lung tissue and blood were collected for protein determination, histological examination and blood biochemistry. The right lung was inflated with 10% buffered formalin at a pressure of 20 cm H2O. Fixed lungs were embedded in paraffin and cut into 4‐μmsections for tissue staining. The left lungs were removed for hydroxyproline evaluation.
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