Participants performed a 3-minute warm-up at 50% of their maximal heart rate (HRmax), as determined during baseline cardiopulmonary exercise testing (ergospirometry), followed by 40 minutes of continuous cycling at 65% of their VO₂max. Sessions were conducted three times per week for a total duration of 8 weeks.

Participants performed a 3-minute warm-up at 50% of their maximal heart rate (HRmax), as determined during baseline cardiopulmonary exercise testing (ergospirometry). The main training protocol consisted of alternating 2-minute cycling bouts: 2 minutes at 65% HRmax followed by 2 minutes at 90% HRmax, repeated continuously for a total of 28 minutes. Sessions were conducted three times per week over an 8-week period.

Percutaneous muscle biopsies of the vastus lateralis (quadriceps) were performed under local anesthesia. Samples were collected at baseline and/or post-intervention to assess microvascular structure, fiber composition, and molecular adaptations to aerobic training.

Muscle tissue samples obtained from the vastus lateralis were processed using optical clearing and immunohistochemical staining techniques to visualize the microvascular network. Confocal microscopy was employed to acquire high-resolution three-dimensional images of arterioles, capillaries, and venules. Quantitative assessments included:

1. Internal diameter measurements, based on endothelial cell boundaries (intima layer)
2. External diameter measurements, based on smooth muscle cells and pericytes (media layer)
3. Capillary density, determined by counting the number of capillaries per sample volume

VO₂max was determined using a graded, incremental exercise test performed on a cycle ergometer until volitional exhaustion. Throughout the test, respiratory gases were continuously measured using a metabolic cart (e.g., COSMED K5), which provided breath-by-breath analysis.

The following parameters were recorded :

1. VO₂ (oxygen uptake) in mL/min
2. VCO₂ (carbon dioxide production) in mL/min
3. Minute ventilation (VE) in L/min