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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

This protocol presents an improved method to obtain transient myocardial hypertrophy with absorbable suture, simulating left ventricular hypertrophy decrease after removing pressure overload. It could be valuable for the studies on myocardial hypertrophic preconditioning.

Abstract

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate cardiomyocyte hypertrophy and slow down progression to heart failure. For novices, however, the MHP model is usually quite difficult to establish because of the technical obstacles in ventilator operation, opening the chest repeatedly, and bleeding caused by debanding. To facilitate this model, to increase the surgical success rate and to reduce the incidence of bleeding, we switched to absorbable sutures for the first TAC combing with a ventilator-free technique. Using a 2-week absorbable suture, we demonstrated that this procedure could cause significant myocardial hypertrophy in 2 weeks; and 4 weeks after surgery, myocardial hypertrophy was almost completely regressed to the baseline. Using this protocol, the operators could master the MHP model easily with a lower operation mortality.

Introduction

Ischemic preconditioning is a phenomenon that induces brief non-lethal episodes of ischemia and reperfusion to the heart and has the capacity to dramatically reduce myocardial injury1. Given the obvious clinical implications of ischemic preconditioning, such as limiting myocardial infarct size2 and suppressing ventricular tachyarrhythmias after myocardial revascularization3, there have been lots of research to dissect the mechanisms underlying cardio-protective effects induced by preconditioning4,5. In contrast, other non-ischemic types of....

Protocol

All procedures were carried out following the guidelines of the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised in 1996). C57BL/6J male mice (8–10 weeks, 20–25 g) were provided by the Animal Center of South Medical University.

1. Preoperative preparation

  1. Pinch off the tip of a 25 G needle with a needle holder and blunt it with a hard object like the holder.
  2. Pass a 5–0 absor.......

Representative Results

In this study, we randomly divided 45 mice into three groups, the sham, the silk suture group, and the absorbable suture group (the number of each group on D0 (baseline), D14, and D28 after TAC was 15, 10, and 5, respectively). On D7, D14, D21, and D28 after the surgery, the constricted peak velocity was determined by echocardiography. We found that the blood flow velocity at the constriction was still greater than 3,000 mm/s in the second week after TAC even though an absorbable suture had been used to constrict the aor.......

Discussion

There is still a vastly underexplored area in cardiac non-ischemic preconditioning. Based on our previous studies, we switched to using absorbable sutures to improve the myocardial hypertrophic preconditioning model.

In previous reports, many investigators used silk suture to constrict the aortic arch8,14,15. Silk suture was easily available and was often used for surgical wound suture, tissue ligatio.......

Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (81770271; to Y, Liao), the Joint Funds of the National Natural Science Foundation of China (U1908205; to Y, Liao), and the Municipal Planning Projects of Scientific Technology of Guangzhou (201804020083; to Dr Liao).

....

Materials

NameCompanyCatalog NumberComments
Absorbable suture (5-0)Shandong Kang Lida Medical Products Co., Ltd5-0Ligation
Animal ultrasound system VEVO2100Visual SonicVEVO2100Echocardiography
Cold light illuminatorOlympusILD-2Light
Heat pad- thermostatic surgical system (ALC-HTP-S1)SHANGHAI ALCOTT BIOTECH COALC-HTP-S1Heating
IsofluraneRWD life scienceR510-22Inhalant anaesthesia
Matrx VIP 3000 Isofurane VaporizerMidmark CorporationVIP 3000Anesthetization
Medical nylon suture (5-0)Ningbo Medical Needle Co.5-0Close the skin
Pentobarbital sodium saltMerck25MGAnesthetization
Precision electronic balanceDenver InstrumentTB-114Weighing sensor
Self-made spacer25-gauge needle
Silk suture (5-0)Yangzhou Yuankang Medical Devices Co., Ltd.5-0Ligation
Small animal microsurgery equipmentNapoxMA-65Surgical instruments
Transmission GelGuang Gong pai250MLEchocardiography
Veet hair removal creamReckitt BenchiserRQ/B 33 Type 2Remove hair of mice
Vertical automatic electrothermal pressure steam sterilizerHefei Huatai Medical Equipment Co.LX-B50LAuto clean the surgical instruments

References

  1. Murry, C. E., Jennings, R. B., Reimer, K. A. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation. 74 (5), 1124-1136 (1986).
  2. Ban, K., et al.

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