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. 2020 Nov 7;5(4):80.
doi: 10.3390/jfmk5040080.

Immunofluorescence Evaluation of Myf5 and MyoD in Masseter Muscle of Unilateral Posterior Crossbite Patients

Giovanna Vermiglio  1 , Antonio Centofanti  1 , Guglielmo Ramieri  2 , Michele Tepedino  3 , Michele Runci Anastasi  4 , Antonio Girolamo Micali  1 , Alba Arco  1 , Maria Grazia Piancino  2
Affiliations

Immunofluorescence Evaluation of Myf5 and MyoD in Masseter Muscle of Unilateral Posterior Crossbite Patients

Giovanna Vermiglio et al. J Funct Morphol Kinesiol. .

Abstract

A unilateral posterior crossbite is a malocclusion where the low activity of the affected masseter muscle is compensated by the contralateral muscle hypertrophy. It is still unknown if, in the same condition, myogenesis with new fibre formation takes place.

Aim: the aim of the present study was to evaluate the expression of myogenesis markers, such as Myf5 and MyoD, in masseter muscles of unilateral posterior crossbite patients.

Materials and methods: biopsies from fifteen surgical patients with unilateral posterior crossbites have been analysed by immunofluorescence reactions. The results show the expression of Myf5 and MyoD in the contralateral muscle but not in the ipsilateral one. Moreover, statistical analysis shows the higher number of satellite cells in the contralateral side if compared to the ipsilateral one.

Conclusions: these results suggest that in contralateral muscle, hyperplastic events take place, as well as hypertrophy.

Keywords: Myf5; MyoD; PAX7; immunofluorescence; masseter muscle; unilateral posterior crossbite.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Images of immunofluorescence reactions performed on contralateral masseter muscle; it is possible to observe satellite cells that co-express both PAX7 (red channel) and Myf5 (green channel) (A,B). (C) sub-figure is an high magnification of PAX7 and Myf-5 positive cells. The yellow asterisk indicates a small group of possibly proliferating PAX7 and Myf-5 positive cells. The yellow fluorescence is obtained from the merge of red and green channels. Nuclei are stained with DAPI (blue channel).
Figure 2
Figure 2
Splitting of channels from Figure 1 with green channel corresponding to Myf5 (A) and red channel corresponding to PAX7 (B). Results show the presence of cells that are positive for PAX7 (B, white arrow) but negative for Myf5 (A, white arrow).
Figure 3
Figure 3
Immunofluorescence reaction in double localization performed on the contralateral masseter muscle. Pictures show muscle fibre characterized by cells that are positive for MyoD (B) but negative for Myf5 (A); these cells start fusing to form young myotubes, characterised by centrally located nuclei (C, dashed line). Nuclei are stained with DAPI (blue channel).
Figure 4
Figure 4
Immunofluorescence reaction in double localization performed on ipsilateral masseter muscle. Results show no Myf5 positive cells (A) and a small number of PAX7 (red channel) positive cells (B); double yellow asterisks indicate PAX7 positive cells. Figure (C) shows the transmitted light.
Figure 5
Figure 5
Immunofluorescence reaction in double localization performed on the ipsilateral masseter muscle showing cells that are negative both for Myf5 (A) and MyoD (B). Nuclei are stained with DAPI (blue channel). The (C) pictures show the transmitted light.
Figure 6
Figure 6
Graphic based on PAX7 positive cells count in 150 fibres, both for ipsilateral and contralateral muscles. Results show a lower number of satellite cells in the ipsilateral muscle if compared to the contralateral one.
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