Experiments were approved by the Austria animal care and use committee and were conform to the “Guide for the Care and Use of Laboratory Animals” published by the US National Institutes of Health (NIH Publication No. 85–23, 1996, revised 2011; available from: www.nap.edu/catalog/5410.html . Hind limb ischemia was induced as described previously18 (link). Briefly, 12–15 weeks old male C57/BL6 mice (Charles River, Sulzfeld, Germany) were administered to anesthesia via an intraperitoneal injection of ketamine hydrochloride (Ketanest, Graeub, Switzerland, 80 mg/kg body weight) and xylazine hydrochloride (Xylasol, aniMedica, Germany, 5 mg/kg body weight). Popliteal artery and femoral artery proximal to the branching into saphenous were ligated with 7–0 polypropylene sutures (Ethicon, USA) and femoral artery was excited subsequently. Limb perfusion was measured using a laser Doppler perfusion image analyser (Moor Instruments, USA). Therefore, animals were kept on a 37 °C tempered heating plate. Limb perfusion was calculated as ration of left (operated, ischemic limb) to right (not operated, non-ischemic limb). For elevation of necrosis upon SWT in vivo, mice were administered to anesthesia as described above. Limbs were treated with SWT subsequently. 15 min after SW application skeletal muscle was harvested for histological processing.
7 0 polypropylene sutures
Manufactured by Johnson & Johnson
Sourced in United States
7-0 polypropylene sutures are a type of surgical suture material made from polypropylene. They are designed for delicate tissue approximation and repair.
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Lab products found in correlation
6 0 polypropylene, by Johnson & Johnson (2 mentions)
C57bl 6 mice, by Charles River Laboratories (1 mentions)
Butorphanol, by Meiji Seika Pharma (1 mentions)
Midazolam, by Astellas Pharma (1 mentions)
Atipamezole, by Nippon Zenyaku Kogyo (1 mentions)
Medetomidine hydrochloride, by Nippon Zenyaku Kogyo (1 mentions)
6 protocols using 7 0 polypropylene sutures
1
Hind Limb Ischemia Induction in Mice
2
Femoral Artery Ligation Ischemic Model
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Anesthesia was administered by an intraperitoneal injection of ketamine hydrochloride (Graeub, Switzerland; 80 mg/kg body weight) and xylazine hydrochloride (aniMedica, Germany; 5 mg/kg body weight). Left femoral artery was ligated and excised between the inguinal ligament and proximal to the branching into saphenous and popliteal artery using 7-0 polypropylene sutures (Ethicon, USA). For analysis, the whole adductor muscle was harvested and split in half resulting in a proximal and distal portion. Of the distal portion, the middle part around the former femoral artery was used for analysis to avoid sampling variances of regions with greater or lesser ischemia.
3
Myocardial Infarction Induction for Ischemic Cardiomyopathy
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Myocardial infarction was induced as it is the standard model for the induction of ischaemic cardiomyopathy. It was performed as described previously 17 . Briefly, animals were anesthetized by an intraperitoneal injection of ketamine hydrochloride (Graeub, Bern, Switzerland; 80 mg/kg bodyweight) and xylazine hydrochloride (aniMedica, Senden, Germany; 5 mg/kg bodyweight). A left lateral thoracotomy was performed in the fourth intercostal space for exposure of the heart. The left anterior descending artery (LAD) was ligated at the level of the pulmonary artery using 7‐0 polypropylene sutures (Ethicon, Somerville, NJ, USA).
4
Allogeneic Endometrial Transplantation in Mice
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Endometrial transplantation was performed as described previously.15 , 27 Briefly, mice were anaesthetized by intraperitoneal (i.p.) injection of a mixture of 0.75 mg/kg medetomidine hydrochloride (Nippon Zenyaku Kogyo Co., Ltd.), 4.0 mg/kg midazolam (Astellas Pharma Inc) and 5.0 mg/kg butorphanol (Meiji Seika Pharma Co., Ltd.). Then, through paravertebral incisions, the mice were bilaterally ovariectomized to exclude the effects of endogenous oestrogen and menstrual cycle. After the procedure, the effect of medetomidine was reversed by i.p. injection of 0.75 mg/kg atipamezole (Nippon Zenyaku Kogyo Co., Ltd.), and the animals were allowed to recover. All donor and recipient mice received subcutaneous injections of estradiol dipropionate (100 mg/kg) in sesame oil (Obahormone Depot, Aska) every week from the time of ovariectomy. Seven days after ovariectomy, the uterine horns from the donor were removed. A round endometrial fragment (3 mm in diameter) was transplanted to each side of the peritoneal wall of recipient mice and secured using 7‐0 polypropylene sutures (Ethicon, Johnson & Johnson). The day of implantation was defined as day 0. WT or RAMP1−/− recipient mice bearing an implant from donor WT or RAMP1−/− mice are referred to hereafter using the terms WT → WT and RAMP1−/− → RAMP1−/−.
5
Evaluating Myocardial Infarction and Surgical Ventricular Restoration in Rats
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Figure 1a shows the experimental protocol. Fifteen 10-week-old male Sprague-Dawley rats weighing 325-368 g were randomized into the following three groups: sham/sham (control group), myocardial infarction (MI)/sham (sham group), and MI/SVR (SVR group), with an interval of 28 days. MI and SVR were performed as previously reported [6] (link). Briefly, in MI, following a left lateral thoracotomy under ventilation, the proximal left anterior descending artery was ligated with 7-0 polypropylene sutures (Ethicon, Somerville, NJ, USA). In SVR, a horizontal sternotomy was performed 28 days after MI, and plication of the akinetic scar area was conducted using three mattress sutures with pledgetted 6-0 polypropylene (Ethicon).
The sham operation was a sternotomy without SVR. Rats were anesthetized with intramuscular ketamine (90 mg/kg) and xylazine (10 mg/kg) for MI and SVR. Rats were euthanized 28 days after SVR (or sham) by intraperitoneal injection of sodium pentobarbitone (150 mg/kg). The excised hearts stopped beating immediately after being soaked in ice-cold phosphate-buffered saline. Histological examination, quantitative realtime reverse transcription polymerase chain reaction (RT-PCR), and western blotting of the left ventricular myocardium were performed. The other 15 rats were used to evaluate the early effects of SVR (2 days) on cardiac function.
The sham operation was a sternotomy without SVR. Rats were anesthetized with intramuscular ketamine (90 mg/kg) and xylazine (10 mg/kg) for MI and SVR. Rats were euthanized 28 days after SVR (or sham) by intraperitoneal injection of sodium pentobarbitone (150 mg/kg). The excised hearts stopped beating immediately after being soaked in ice-cold phosphate-buffered saline. Histological examination, quantitative realtime reverse transcription polymerase chain reaction (RT-PCR), and western blotting of the left ventricular myocardium were performed. The other 15 rats were used to evaluate the early effects of SVR (2 days) on cardiac function.
6
Rat Model of Left Ventricular Plication
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General anesthesia was induced by intramuscular ketamine (90 mg/kg) and xylazine (10 mg/kg). Following left lateral thoracotomy under ventilation, the proximal LAD was ligated with 7-0 polypropylene sutures (Ethicon, Somerville, New Jersey). The MI at the anterior LV wall was confirmed by akinesis through an echocardiographic examination before the chest was closed. Four weeks after the LAD ligation, the rats developed a large scar, but not an aneurysm, on the anterior LV wall with hypokinesis in the non-ischemic lesion (Video 1).
LVP was performed as reported previously under the same anesthesia as the MI. 9 Briefly, after ligation of the internal mammary arteries, a horizontal sternotomy was performed at the level of the fifth intercostal space to expose the entire heart. Thereafter, LVP was performed by plicating the akinetic scar area using three mattress sutures with pledgetted 6-0 polypropylene (Ethicon). Much attention was paid not to stitch on the scar tissue but on the viable myocardium. An over-and-over suture was then performed on the plicated scar tissue (Video 1).
LVP was performed as reported previously under the same anesthesia as the MI. 9 Briefly, after ligation of the internal mammary arteries, a horizontal sternotomy was performed at the level of the fifth intercostal space to expose the entire heart. Thereafter, LVP was performed by plicating the akinetic scar area using three mattress sutures with pledgetted 6-0 polypropylene (Ethicon). Much attention was paid not to stitch on the scar tissue but on the viable myocardium. An over-and-over suture was then performed on the plicated scar tissue (Video 1).
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