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.

Prevalence of hypodontia and supernumerary teeth in a German cleft lip with/without palate population.

BACKGROUND: The distribution of dental abnormalities among cleft patients concerning cleft type frequently poses ambiguity wherefore the aim of this study was to evaluate the prevalence of hypodontia and supernumerary teeth in an exemplary German cleft population dependent on the cleft type. METHODS: Radiographs and dental records of cleft patients, which had been treated and followed up in the Department of Oral and Maxillofacial Surgery, University Hospital Carl Gustav Carus Campus, Dresden, Germany (investigation period of 22 years) were evaluated concerning hypodontia and supernumerary teeth dependent on the cleft type. Out of 386 records, 108 patients met the inclusion criteria: non-syndromic cleft of the alveolus with or without palate (CL/P), at least one clear panoramic x-ray, sufficient dental records. Statistical analysis was performed using x-square and binominal test (p ≤ 0.05). RESULTS: Hypodontia was more frequent (54/50%) than supernumerary teeth (36/33.3%) and was more common in bilateral clefts of the lip and palate (BCLP) (70.1%) than in unilateral clefts of the lip and palate (UCLP) (51.6%) or clefts of the lip and alveolus (CLA) (34.5%) (p << 0.001). There was an average of 0.9 missing teeth per patient, thereof the upper lateral incisor was most often affected (23.2%). In contrast, supernumerary teeth were more frequent in CLA (51.7%; p = 0.014) than UCLP (29.0%) and BCLP patients (17.6%). CONCLUSION: The prevalence for numerical dental anomalies was significantly different among the cleft types. Hypodontia significantly increased with the extend of the cleft, whereas the prevalence of supernumerary teeth decreased.

Histological features of masticatory muscles after botulinum toxin A injection into the right masseter muscle of dystrophin deficient (mdx-) mice.

OBJECTIVE/BACKGROUND: The most frequently used animal model for human DMD (Duchenne muscular dystrophy) research is the mdx mouse. In both species, characteristic histological changes like inflammation, muscle fiber degeneration and fibrosis are the same, but in contrast to humans, in mdx mice, phases of muscle fiber degeneration are compensated by regeneration processes. AIM: Therefore, the interest of this study was to evaluate histological features in masticatory muscles after BTX-A injection into the right masseter muscle of wild type and dystrophic (mdx) mice, illustrating de- and regeneration processes induced by this substance. MATERIAL AND METHODS: The right masseter muscle of 100 days old healthy and mdx mice were selectively paralyzed by a single intramuscular BTX-A injection. Masseter as well as temporal muscle of injection and non-injection side were carefully dissected 21 days and 42 days after injection, respectively, and fiber diameter, cell nuclei position, necrosis and collagen content were analyzed histomorphologically in order to evaluate de- and regeneration processes in these muscles. Statistical analysis was performed using SigmaStat Software and Mann Whitney U-test (significance level: p < 0.05). RESULTS: At both investigation periods and in both mouse strains fiber diameter was significantly reduced and collagen content was significantly increased in the right injected masseter muscle whereas fiber diameters in mdx mice were much smaller, and these differences were even more apparent at the second investigation period. Necrosis and central located nuclei could generally be found in all mdx mice muscles investigated with an amount of centronucleation exceeding 60%, and a significant increase of necrosis six weeks after injection. In wild type mice central located nuclei could primarily be found in the treated masseter muscle with a portion of 2.7%, and this portion decreased after six weeks, whereas in mdx mice a decrease could also be seen in the non-injected muscles. In contrast, in wild type mice necrosis was not apparent at any time and in all muscles investigated. CONCLUSION: From our results it can be concluded that in mdx mice masticatory muscles de- and regeneration processes were extended, triggered by a selective BTX-A injection, or mdx mice at this age, independently of BTX-A treatment, went through another cycle of de- and regeneration as a characteristic of this disease.

Influence of Botulinumtoxin A on the Expression of Adult MyHC Isoforms in the Masticatory Muscles in Dystrophin-Deficient Mice (Mdx-Mice).

The most widespread animal model to investigate Duchenne muscular dystrophy is the mdx-mouse. In contrast to humans, phases of muscle degeneration are replaced by regeneration processes; hence there is only a restricted time slot for research. The aim of the study was to investigate if an intramuscular injection of BTX-A is able to break down muscle regeneration and has direct implications on the gene expression of myosin heavy chains in the corresponding treated and untreated muscles. Therefore, paralysis of the right masseter muscle was induced in adult healthy and dystrophic mice by a specific intramuscular injection of BTX-A. After 21 days the mRNA expression and protein content of MyHC isoforms of the right and left masseter, temporal, and the tongue muscle were determined using quantitative RT-PCR and Western blot technique. MyHC-IIa and MyHC-I-mRNA expression significantly increased in the paralyzed masseter muscle of control-mice, whereas MyHC-IIb and MyHC-IIx/d-mRNA were decreased. In dystrophic muscles no effect of BTX-A could be detected at the level of MyHC. This study suggests that BTX-A injection is a suitable method to simulate DMD-pathogenesis in healthy mice but further investigations are necessary to fully analyse the BTX-A effect and to generate sustained muscular atrophy in mdx-mice.

Radiographic Study of the Prevalence and Distribution of Hypodontia Associated with Unilateral and Bilateral Clef Lip and Palate in a Hungarian Population.

BACKGROUND Cleft defects are one of the most frequent birth-deformities of the orofacial region and they are commonly associated with anomalies of the tooth structure, size, shape, formation, eruption, and tooth number. The aim of our study was to evaluate the prevalence, distribution, and potential association of combined hypodontia in cleft-affected patients with regard to all types of teeth in both jaws in the permanent dentition. MATERIAL AND METHODS This retrospective radiographic analysis included patients with various types of clefts treated orthodontically in the Department of Orofacial Orthopedics and Orthodontics at Heim Pàl Children's Hospital, Budapest. There were 150 patients (84 males, 66 females) with non-syndromic unilateral (UCLP; n=120 patients) or bilateral (BCLP; n=30 patients) cleft formation (lip, alveolus and palate) who met the inclusion criteria. Statistical analysis was performed using the chi-square test and Fisher's exact test (significance level p<0.05). RESULTS Hypodontia was significantly more frequent in patients with cleft-sided lateral incisor (104 patients, 69%), with a total of 235 missing teeth, followed by the second premolars of the upper and lower jaw. A significant correlation of congenital missing teeth was observed in left-sided clefts between the upper and lower second premolar in the cleft area CONCLUSIONS Hypodontia inside and outside the cleft area was frequently observed. This should affect the therapy plans, especially if the cleft-sided premolar is also absent. Further comprehensive research including numerous random samples is necessary for better estimating other possible associations.

Mandibular midline distraction osteogenesis.

In orthodontics, bone structure, its density and dimensions play an essential role by explaining limitations in magnitude, size and extent of tooth movement. Severe anterior crowding is one of the most frequently encountered dental malocclusions. Its therapy is mostly limited by lack of basal and alveolar bone and it often involves tooth extractions. Mandibular midline distraction osteogenesis is a method of natural bone generation and also a treatment option to achieve space regaining in a much-reduced lower jaw with distinctive frontal place deficit and severe anterior crowding, without sacrificing permanent teeth. McCarthy and Guerrero were of the first researchers reporting on this method applied on human lower jaws and they increased clinical interest in this approach. Although this method has been clinically used ever since, many questions concerning effects on bone regeneration speed, bone quality, tooth movement into regenerated area, periodontal health and long-time stability of treatment outcomes have not been sufficiently investigated. This overview should present the current clinical and biological state of knowledge about bone gain and tooth movement through regenerate bone. Furthermore it should encourage interest in further research on this topic.