Age-related alterations of hyaluronan and collagen in extracellular matrix of the muscle spindles

Background: Muscle spindles (MSs) play a crucial role in proprioception and locomotor co-ordination. Although the elasticity and viscosity of the extracellular matrix (ECM) within which MSs are embedded may play a key role in MS function, the impact of aging on ECM components is unclear. The aim of the current study was to investigate the age-related physiological changes of the ECM and to verify if these could be due to alterations of the environment directly surrounding MSs. Methods: Hematoxylin Eosin and picrosirius-red staining was carried out; collagen types I (COLI) and III (COLIII) were assessed, and biotinylated hyaluronan binding protein (HABP) immunohistochemical analysis was undertaken to evaluate alterations of the ECM in the intramuscular connective tissue (IMCT) of the hindlimbs of C57BL/6J male mice. Assessments were carried out on 6-week-old (Group A), 8-month-old (Group B), and 2-year-old (Group C) laboratory mice. Results: The capsule’s outer layer became progressively thicker with aging (it was 3.02 ± 0.26 µm in Group A, 3.64 ± 0.31 µm in Group B, and 5.81 ± 0.85 µm in Group C). The collagen in IMCT around and within the MSs was significantly higher in Group C, but there were no significant differences between Groups A and B. The MS capsules and continuous IMCT were primarily made up of COLI and COLIII. The average optical density (AOD) values of COLI in IMCT surrounding MS were significantly higher after aging (p < 0.05), but there were no significant differences in COLIII in the three groups (p > 0.05). HA was present in IMCT and filled the MSs capsule. The AOD of HABP of MS showed that there were lower HA levels in Group C with respect to Group A (p = 0.022); no significant differences were noted neither between Groups A and B nor between Groups B and C (p > 0.05). Conclusion: Age-related collagen accumulation and lower HA in the ECM in which the MSs were embedded may probably cause more stiffness in the ECM in vivo, which could help to partly explain the peripheral mechanisms underlying the age-related decline in functional changes related to MSs.