Effect of age on fatty infiltration of supraspinatus muscle after experimental tendon release in rats

Abstract

Background: Rotator cuff tendon tear is a leading cause for atrophy, fibrosis and fatty infiltration of the rotator cuff muscles. The pathophysiology of fatty muscle infiltration is not well understood. An animal model suited to study cellular and molecular mechanisms would therefore be desirable. While a rat model has been established for chronic rotator cuff tendon pathology, sufficient and easily identifiable fatty infiltration of the muscle has not yet been shown in rats. As younger animals regenerate better, we hypothesized that the absence of a sufficient amount of fatty infiltration in previous experiments was due to the selection of young animals and that older animals would exhibit higher amounts of fatty infiltration after tendon tear.

Findings: The supraspinatus tendon was released using tenotomy in 3 young (6 weeks old) and in 3 aged (24 months old) Sprague Dawley rats (group I and II). Another 3 aged (24 months old) rats underwent sham surgery and served as a control group (group III). In group I and II retraction of the musculotendinous unit was allowed for 6 weeks. All animals were sacrificed 6 weeks after surgery and the supraspinatus muscles were harvested. Each sample was examined for fatty infiltration of the muscle by histological methods and micro-CT. In histology, fat cells were counted with a 10-fold magnification in 6 fields of view twice. An adjusted measurement setup was developed for the use of micro-CT to quantify the absorption coefficient of the muscle as a reciprocal indicator for fatty infiltration, based on the established procedure for quantification of fatty infiltration on CT in humans.Tenotomy resulted in an insignificant increase of fat cells in histological sections in both, aged and young rats. Micro-CT was able to quantify small differences in the absorption coefficients of muscle samples; the absorption coefficient was 8.1% ± 11.3% lower in retracted muscles (group I and II) compared with the control (group III), indicating a tendency towards a higher amount of intra- and/or extracellular fat. Absorption was 4.28% ± 3.2% higher in aged compared with young muscles; however, this difference could not be confirmed by histology.

Conclusions: Substantial fatty muscle infiltration following chronic retraction after supraspinatus tenotomy could be documented histologically neither in young nor aged rats. Although micro-CT was able to reveal minor differences in absorption between the studied groups, the differences were too small to make the rat supraspinatus model interesting to study fatty infiltration of the chronically retracted muscle.

Figure 1

Fat cells in histological sections. Fat cells (arrows) were found 6 weeks after tendon release in muscles of aged animals (left) in a small amount and less frequently in the control group that did undergo sham surgery (right).

Figure 2

Micro-CT values. The absorption coefficient of the muscle measured by micro-CT in mu (1/mm) as a reciprocal indicator for fatty infiltration was higher in group I (0.93 ± 0.01, aged rats, tenotomy) than group group II (0.89 ± 0.01, young rats, tenotomy) but lower than in group III (0.99 ± 0.05, aged rats, sham surgery).