Density of nerve fibres and expression of substance P, NR2B-receptors and nerve growth factor in healthy human masseter muscle: An immunohistochemical study

Abstract

Background: In skeletal muscle, free nerve endings are mostly located within the connective tissue. However, the distribution of sensory afferent fibres in healthy human masseter muscle tissues has not been studied.

Objectives: Primarily to investigate human masseter muscle nerve fibre densities as well as expression of NR2B receptors, substance P (SP) and nerve growth factor (NGF), and secondarily to compare this between a) nerve fibres associated with myocytes and within connective tissue; b) sexes; and c) ages.

Methods: Microbiopsies of the masseter muscle were obtained from 60 sex- and age-matched healthy participants. Biopsy sections were analysed using immunohistochemistry and were visualised with a Leica TCS SPE confocal microscope. The Mann-Whitney U test was used for statistical analyses.

Results: The density of nerve fibres within connective tissue was significantly greater than in nerve fibres associated with myocytes (P < .001). Nerve fibres within connective tissue expressed SP alone or together with NR2B significantly more often than those associated with myocytes (P < .001). The frequency of nerve fibres, which expressed SP alone or in combination with NR2B or NGF, was significantly greater in women than in men (P < .050). Moreover, the co-expression of the three markers together was inversely correlated with age in women (P < .002).

Conclusions: There is a higher density and greater expression of sensory nerve fibres within the connective tissue than associated with myocytes in healthy human masseter muscle. This suggests that nerve fibres within connective tissue are more involved in nociception than nerve fibres associated with myocytes.

Keywords: NR2B; expression; immunohistochemistry; muscle biopsy; nerve growth factor; substance P.