Fluorescent Microspheres Injected into Skeletal Muscle to Observe Fluid Movement in Interstitial Space

Abstract

The force within a muscle may be assessed using intramuscular pressure measurements (IMP). However, a limiting factor in implementing this technology in the clinic is the observed variability of IMP measurements. The variability may be due in part to the flow of interstitial fluid. The objective of this study was to develop a technique for tracking interstitial fluid flow during muscle lengthening using fluorescent microspheres. Additionally, the effect of microsphere diameter on dispersion was evaluated. Fluorescent microspheres were injected into the rat tibialis anterior (TA) either before (right limb) or after (left limb) lengthening to 10% strain (n = 5). To help discriminate the effects of microsphere size, 10 µl of microspheres (0.25% by volume) were injected simultaneously: 0.1 µm in sky blue and 0.2 µm in yellow green. Once frozen, 20 µm muscle cross‐sections were stained and imaged using confocal microscopy to evaluate the microsphere's axial dispersion as a percent of muscle length. The microsphere dispersion was normalized to the muscle length. The fluorescent microspheres flowed with the interstitial fluid and did not penetrate the muscle fiber. There was significantly greater microsphere dispersion with passive strain (p‐value > 0.01) for lengthened samples (31±7%) compared to static samples (17±9%). Dispersion did not differ for the two bead sizes. In conclusion, this illustrates the microspheres track the interstitial fluid and passive lengthening results in additional dispersion.Supported by NIH under award number R01HD31476 and T32HL105355.