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This murine model combines a septic insult with hindlimb muscle disuse to recapitulate the bedridden feature of the typical septic patient. The model represents a significant departure from previous models to study muscle dysfunction in sepsis and is a reproducible approach to addressing therapeutic strategies to treat this condition.
Sepsis is a major cause of in-hospital deaths. Improvements in treatment result in a greater number of sepsis survivors. Approximately 75% of the survivors develop muscle weakness and atrophy, increasing the incidence of hospital readmissions and mortality. However, the available preclinical models of sepsis do not address skeletal muscle disuse, a key component for the development of sepsis-induced myopathy. Our objective in this protocol is to provide a step-by-step guideline for a mouse model that reproduces the clinical setting experienced by a bedridden septic patient. Male C57Bl/6 mice were used to develop this model. Mice underwent cecal ligation and puncture (CLP) to induce sepsis. Four days post-CLP, mice were subjected to hindlimb suspension (HLS) for seven days. Results were compared with sham-matched surgeries and/or animals with normal ambulation (NA). Muscles were dissected for in vitro muscle mechanics and morphological assessments. The model results in marked muscle atrophy and weakness, a similar phenotype observed in septic patients. The model represents a platform for testing potential therapeutic strategies for the mitigation of sepsis-induced myopathy.
Sepsis is a life-threatening condition due to an overactive immune response that adversely affects multiple organ systems, resulting in a major burden to health systems worldwide1. More recently, the in-hospital mortality linked to sepsis has decreased due to improved intensity care unit (ICU) management1,2. However, approximately 75% of the patients surviving the initial septic insult develop skeletal muscle atrophy (e.g., reductions in cross-sectional area) and weakness (e.g., reductions in force production capacity)3,4. This ....
The procedures have been reviewed and approved by the University of Florida IACUC (#202200000227). Male C57BL/6J mice, 17 weeks old, with a body mass ranging from 27 g to 34 g, were used for the present study. The experimental procedures and timeline outlined in this protocol are depicted in Figure 1. As indicated, the protocol spans a total of 11 days. Animals undergo survival surgery (CLP/Sham) on Day 0, followed by four days of fluid and analgesic support. On Day 4, animals begin HLS for .......
For the representative data shown in the results, male C57BL/6J mice, 17 weeks old, with a body mass ranging from 27 to 34 g, were used. The entire protocol takes eleven days to complete and consists of the surgery intervention (CLP or sham), saline and analgesic support (days 0 to 4), and the HLS disuse (days 4 to 11). Terminal experiments can be performed at any point over the suspension phase. To better understand the impact of the model on skeletal muscle function, the results are compiled from several experiments ac.......
The current protocol provides technical guidelines for the implementation of a new preclinical model of sepsis-induced myopathy. All materials and important steps are described in detail for the reproduction of the model. This approach can reproduce the skeletal muscle dysfunction observed in septic patients, highlighting the role of disuse as a crucial component in worsening myopathy. Thus far, the majority of the preclinical studies addressing sepsis-induced myopathy did not include the disuse as an active component of.......
This work was supported by NIH R21 AG072011 to OL.
....Name | Company | Catalog Number | Comments |
4-0 Ethicon Coated Vicryl | Ethicon | D5792 | Absorbable suture used for closure of muscle layer and for ligation of the cecum. |
4-0 Ethilon Black 18" | Ethicon | 662G | Non absorbable suture for closure of the skin layer. |
BD PrecisionGlide Needle 26-28 G | BD | 305136 for 27g needle | Needle for puncturing the cecum. |
C57BL/6J mice | Jackson Laboratory | strain #000664 | |
Cotton Tipped Applicators | Puritan | S-18991 | Swabs for topical application of iodine. |
Cryostat | (Leica CM1950) | ||
Dynarex Povidone Iodine Prep Solution | Dynarex | 1415 | Topical Antiseptic Liquid for Skin and Mucosa |
Ethanol 200 Proof (100%) | Fisher Scientific | To make 70% ethanol for cleaning skin. | |
Hindlimb Suspension Cages | Custom Made | N/A | These custom made cages will be highlighted in the video recordings of the MS. |
Optixcare Eye Lube | Optixcare | Eye lube for protection during survival surgery. | |
Scalpel blades #11 | Fine Science | Blade used to make incisions on skin and muscle. | |
Skin-Trac | Zimmer | 736579 | Foam tape for fixing the tail to the suspension apparatus. |
SomnoSuite Low-Flow Digital Vaporizer | Kent Scientific Corporation | SS-01 | Vaporizer for Isoflurane Anesthesia |
Tissue bath apparatus | Aurora Scientific | Model 800A, Dual Mode Muscle Lever 300C |
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