Influence of botulinum toxin A on craniofacial morphology after injection into the right masseter muscle of dystrophin deficient (mdx-) mice

Background: Severe craniofacial and dental abnormalities, typical for patients with progressive Duchenne muscular dystrophy (DMD), are an exellcent demonstration of Melvin L. Moss "functional matrix theory", highlighting the influence of muscle tissue on craniofacial growth and morphology. However, the currently best approved animal model for investigation of this interplay is the mdx-mouse, which offers only a limited time window for research, due to the ability of muscle regeneration, in contrast to the human course of the disease. The aim of this study was to evaluate craniofacial morphology after BTX-A induced muscle paralysis in C57Bl- and mdx-mice, to prove the suitability of BTX-A intervention to inhibit muscle regeneration in mdx-mice and thus, mimicking the human course of the DMD disease.

Methods: Paralysis of the right masseter muscle was induced in 100 days old C57Bl- and mdx-mice by a single specific intramuscular BTX-A injection. Mice skulls were obtained at 21 days and 42 days after BTX-A injection and 3D radiological evaluation was performed in order to measure various craniofacial dimensions in the sagittal, transversal and vertical plane. Statstical analysis were performed using SigmaStat®Version 3.5. In case of normal distribution, unpaired t-test and otherwise the Mann-Whitney-U test was applied. A statistical significance was given in case of p ≤ 0.05.

Results: In contrast to C57Bl-mice, in mdx-mice, three weeks after BTX-A treatment a significant decrease of skull dimensions was noted in most of the measurements followed by a significant increase at the second investigation period.

Conclusions: BTX-A can induce changes in craniofacial morphology and presumably partially inhibit muscle regeneration in mdx-mice, but cannot completely intensify craniofacial effects elicited by dystrophy. Further research is necessary in order to fully understand muscle-bone interplay after BTX-A injection into dystrophic muscles.