Evaluation of the Regeneration Capacity of Satellite Cells From the Quadriceps Compared to That of the Diaphragm

NCT05194631 | Completed

• 1 other identifier: RECHMPL21_0384
• Study Type: interventional
• Target: 12
• Locations: 1 country
• Sites: 1

Brief Summary

Actually no treatment exists to prevent the loss of diaphragmatic function induced by mechanical ventilation during an intensive care unit stay. The consequence is a growing number of survivors with moderate to severe chronic respiratory disease so called Ventilation-Induced Diaphragmatic Dysfunction (VIDD). This study complements the BOTAN study (IRB Accreditation number: 198711, NCT02900300) on 24/11/2021 under number 2021\_IRB-MTP\_08-37), which aim was the constitution of a biological bank of diaphragmatic tissue in order to be able to propose an innovative tool for the in vitro screening of bioactive molecules of interest in human, to improve the regenerative capacity of this main and essential muscle for breathing. In this present study, in order to offer this evaluation tool in the screening of bioactive molecules of interest, without the need of the diaphragm biopsy, but simply of the quadriceps, the investigators propose, by adding a quadriceps muscular biopsy to patients already included in BOTAN study, to compare the regenerative capacity of quadriceps and diaphragmatic muscle in the same individual. Thus, this study is the first, which will propose a new minimally invasive tool for the evaluation and optimization of future pharmacological treatments targeted to limit the impact of VIDD. Thus our main objective of thus study is to compare in the same patient the regenerative capacity of the muscular fiber of the quadriceps to that of the diaphragm in order to set up an in vitro model allowing to test in humans future pharmacological treatment devoted to limit the incidence of VIDD. In this study will be recruited patients treated surgically for a benign or malignant tumor of the liver requiring surgical excision in contact with the diaphragm and who have agreed to participate in the BOTAN study. The resection to be complete (R0) must pass within the diaphragmatic muscle allowing us to obtain without any constraint for the patient, diaphragmatic tissue. The provision of 10 samples will enable us to produce the cell cultures needed to achieve the study objectives. In order to take into account subjects who leave the study without meeting the primary endpoint, we plan to include up to 15 subjects. During this surgery, for the purposes of the present study, the investigators will perform a quadriceps biopsy. Quadriceps biopsies are widely used in biomedical research and no deleterious effects have been described. The surgical team is fully qualified for this type of sampling and all experiments necessary to compare regenerative capacity of quadriceps and diaphragmatic muscular fiber use laboratory techniques perfectly mastered by our research team. For each patients, this study will be performed over two visits. The first, during a consultation will allow the inclusion of the patient, as well as the collection of clinical data and a second during the scheduled surgical intervention where quadriceps biopsy will be realised. The expected duration of inclusions will be one year. No immediate individual benefit is expected for the patient. This study aims to improve, in the future, therapies that reduce the incidence of diaphragmatic dysfunction induced by mechanical ventilation. In fact, diaphragmatic dysfunction induced by ventilation is observed in at least 50% of patients in intensive care; this dysfunction has been implicated in the significant lengthening of the hospital stay, the difficulties in weaning from mechanical ventilation and in the increased risk of co-morbidity and mortality. The validation of the first in vitro model in humans of this dysfunction, from quadriceps biopsy, will allow us to pre-select antioxidant molecules and the most effective concentrations to use for a future therapeutic trial aimed at limiting the deleterious effects of mechanical ventilation on the diaphragm in humans.

Conditions

Diaphragm Injury; Mechanical Ventilation Complication

Keywords

Satellite cells; mechanical and oxidative stress; regenerative function

Outcome Measures

Primary Outcomes (1)

• Comparison of the number and size of differentiating myoblasts obtained per satellite cells from the quadriceps with that of the diaphragm in the same patient

  The quadriceps and diaphragm muscle biopsies will be placed in a culture dish treated with type 1 collagen in a thin layer of matrigel. 6 to 8 days later, the myoblasts are purified by MACS (Magnetic Activated Cell Sorting) based on the property of myoblasts to express the CD56 surface antigen. After culture on a collagen dish, proliferation and differentiation of the CD56 positive cells will be assessed by cell counting and immunofluorescence with antibodies directed against muscle markers.

  3 months

Secondary Outcomes (3)

• Elongation of muscle fibre length in a human model of ventilation-induced diaphragmatic dysfunction (VIDD) using myoblasts from the diaphragm and quadriceps

  3 months

• Evaluation of mechanical stresses of muscle fibres from the quadriceps and the diaphragm in a human model of ventilation-induced diaphragmatic dysfunction (VIDD)

  3 months

• Evaluation of the in vitro efficacy in humans of a mitochondria-specific anti-oxidant (SS-31) to prevent a deficit in muscle regeneration induced by oxidative stress

  3 months

Study Arms (1)

Regeneration Capacity of Satellite Cells

EXPERIMENTAL

In vitro Regeneration Capacity of Satellite Cells from the Quadriceps Compared to That of the Diaphragm isolated From one patient

Other: Quadriceps microbiopsy using a Tru-Cut biopsy needle

Interventions

Quadriceps microbiopsy using a Tru-Cut biopsy needle OTHER

During surgery for a benign or malignant tumor of the liver a quadriceps microbiopsy will be performed. A 1 cm incision of the skin and fat planes will be made using a scalpel until contact with the quadriceps aponeurosis. The biopsy needle will be inserted superficially into the muscular part allowing, through a suction system, a micro biopsy of the quadriceps of approximately 25 mg.

Regeneration Capacity of Satellite Cells

Eligibility Criteria

• Age: 30 Years - 80 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• age between 30 and 80 years old
• patient treated by the digestive surgery department of Montpellier University Hospital for a benign or malignant tumor of the liver requiring surgical excision in contact with the diaphragm who agreed to participate in the BOTAN study (Approval Number assigned by the IRB of Montpellier : 2021\_IRB-MTP\_08-37)
• patient clinically stable at the time of the study, ie not requiring any treatment

You may not qualify if:

• Patients undergoing treatment with antibiotics or corticosteroids, or recently within the last 4 months
• Pregnant or breastfeeding woman.
• Patients with a body mass index \> 30
• Patients with any criteria that may in themselves impair respiratory muscle function such as chronic obstructive pulmonary disease, heart failure, systemic infection, neuromuscular pathology, psychiatric pathology or metabolic disorder.
• Patients with coagulopathy or thrombocytopenia.

Sponsors & Collaborators

• University Hospital, Montpellier: lead

Study Sites (1)

University Hospital

Montpellier, 34295, France

Location

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NA
• Masking: NONE
• Purpose: BASIC SCIENCE
• Intervention Model: SINGLE GROUP
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: This is a prospective cross-sectional study involving category humans, which aims is to assess the pathophysiological mechanisms of a deficit in muscle fiber regeneration

Study Record Dates

• First Submitted: December 9, 2021
• First Posted: January 18, 2022
• Study Start: February 18, 2022
• Primary Completion: May 30, 2024
• Study Completion: May 30, 2024
• Last Updated: December 6, 2024

Record last verified: 2023-08

Data Sharing

• IPD Sharing: Will not share

Locations

FR (1)