In-vitro human whole blood clots, approximately 200–300 μm in diameter, comparable to intracerebral segments of the middle cerebral arteries were prepared. [48 (link), 49 , 35 (link)]. Briefly, whole blood was obtained from human volunteers by sterile venipuncture after Institutional Review Board (IRB) approval of the protocol and written informed consent. Aliquots were placed into 1.5 mm diameter micropipettes and allowed to clot around 7-0 silk sutures (Ethicon) at room temperature for approximately 10 minutes. The clots were incubated at 37° C for 3 hours in a temperature-controlled water bath. This method yielded sample clots that were 230 ± 34 μm in diameter. The clots were stored at 4° C in a humidified environment to ensure platelet viability. [50 (link)]
7 0 silk suture
Manufactured by Johnson & Johnson
Sourced in Germany, United Kingdom
The 7-0 silk suture is a non-absorbable, sterile, braided surgical suture made from natural silk fibers. It is designed for use in fine suturing procedures, such as in ophthalmology or microsurgery, where precise wound closure is required.
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7 protocols using 7 0 silk suture
1
In-vitro Human Blood Clot Formation
2
Murine Myocardial Infarction Model via LAD Ligation
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Myocardial infarction surgery was performed on 2-month-old C57BL/6 mice by ligation of the left anterior descending (LAD) coronary artery [60 (link)]. Mice were anesthetized with isoflurane. After the chest was shaved and cleaned with 75% alcohol, a suture was placed around the front upper incisors and pulled taut so that the neck was slightly extended. For oral intubation, the tongue was retracted and held with forceps, and a 20G catheter was inserted into the trachea. The catheter was then attached to the mouse ventilator via a Y-shaped connector. Ventilation was performed with a tidal volume of 225 µL for a 25-g mouse and a respiratory rate of 130 breaths per minute. 100% oxygen was provided to the inflow of the ventilator. Then the chest was opened through a left parasternal incision, and the heart exposed at the left 3rd–4th intercostal space. Chest retractor was applied to facilitate the view. The pericardium was opened, and the ligation was performed on the LAD coronary artery using 7–0 silk sutures (Ethicon). The lungs were slightly overinflated to assist in removal of air in the pleural cavity. Dissected intercostal space and chest skin were closed using a 6–0 silk suture (Ethicon). The sham group underwent the same surgical procedure (except that the LCA was not occluded).
3
Cardiac Ischemia and Monocyte Analysis in Mice
4
Mouse Model of Myocardial Infarction
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Mouse MI surgery was performed as previously described.18 (link) Briefly, mouse was anaesthetized with 1.5% isoflurane, orally intubated, and then connected to a rodent ventilator. After removing the fur and opening the left thorax, the heart was exposed and MI was produced by permanent ligation of the left anterior descending (LAD) artery using a 7-0 silk suture (Ethicon, Somerville, NJ). Bleaching of the distal myocardium verified ischemia. Sham-operated mice underwent the same procedure without LAD ligation. Mice within 1 to 10 days of operation were used to analyze heart immune cell infiltration. Mice at 1, 7, and 28 days post-MI 30 days were used to analyze the heart function with a pre-harvest echocardiogram being performed for reference. Hearts were used for real-time PCR, immunoblot, and immunohistological or immunofluorescent staining.
5
Neonatal Hypoxic-Ischemic Brain Injury Model
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PND5 and PND9 pups were exposed to neonatal HI as previously described (21 (link), 22 (link)). Pups were anesthetized with isoflurane (IsoFlo vet 100%; Abbott Laboratories Ltd, Illinois, USA), 5% for induction and 1.5% for maintenance, in 1:1 oxygen-nitrogen gas mixture. The left common carotid artery was ligated with a 7.0 silk suture (Ethicon; Vömel, Germany) and the incision was closed and infiltrated with a local anesthetic (Xylocain 20 mg/ml, lidocaine hydrochloride; Astra Zeneca, Södertälje, Sweden). After surgery, mice were returned to their dams for 1 h, before being placed in a chamber with circulating humidified air (36 °C). In the chamber, PND5 mice were exposed to 10 min of air, followed by 60 min of hypoxia (10% O2 in 90% N2) followed by another 10 min of air. PND9 mice were treated in the same manner except the hypoxia time was 50 min. This procedure results in a similar degree of brain injury in PND5 and PND9 mice as previously shown (21 (link), 23 (link)) and Supplementary Figure 1 . Following the hypoxic exposure, pups were returned to their dams until sacrifice.
6
Sciatic Nerve Excision in Animal Model
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Animals were prepared for surgery with a subcutaneous injection of Vetergesic (0.05 mg kg−1 Alstoe-Sogeval UK Ltd.). They were then anaesthetized by inhalation of isoflurane and oxygen (2–5%, 0.4–1 L/min; Merial Animal Health, Harlow, UK). The area surrounding the sciatic notch was shaved and wiped with a sterilizing solution. A small incision was made, and then blunt dissection was used to expose the sciatic nerve. A small section (~2 mm) of the nerve was removed. The wound was then closed with 7/0 silk suture (Ethicon, Livingston, UK).
7
Induction of Myocardial Infarction in Mice
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Myocardial infarctions were produced as described previously Xiao et al. (2011 (link)). In brief, mice were anesthetized by intramuscular injection with a mixture of ketamine (9 mg/100 g), Acepromazine (4 mg/100 g), atropine (0.06 mg/100 g), and prepared for sterile surgery. Animals were intubated with a 20G catheter and ventilated with a mixture of O2 and room air, using a model 845 ventilator (Harvard Apparatus, Holliston, MA). The stroke volume was set at 0.3 mL and the respiratory rate was 125 breaths/min. After a left anterior thoracotomy, the heart was exposed and the location of the left coronary artery (LCA) on the surface of LV anterior wall was identified. A 7–0 silk suture (Ethicon) was placed around the LCA and was tightened. Occlusion of the LCA was confirmed by change in the color at the involved LV wall. Lungs were expanded to displace air before the chest was closed; the mouse was then extubated. Mice were allowed to recover from the surgery in an oxygen‐filled chamber over a heating pad. Sham‐operated mice underwent similar surgery without occlusion of the LCA.
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