Structural characteristics of periodontal ligaments in the pig model with experimental periodontitis.

To investigate how the structure of Sharpey's fibers in the periodontal ligaments (PDL) were affected by experimental periodontitis in a young pig model, 7 were periodically inoculated with four types of bacteria and a ligature around the last maxillary molar for 8 weeks to induce periodontitis (PG), and 10 served as controls (CG). The harvested molar blocks were sectioned coronally and stained with either hematoxylin & eosin (H&E) or Sirius Red (SR). The H&E-stained images were first reviewed. Then, images of each adjacent SR-stained sections were captured at the region close to the apex of mesial roots under polarizing light microscopy. Sharpey's fiber numbers in each bundle, total bundle numbers, connection of the bundle to the cementum and alveoli, as well as bundle angulations to the molar long axis were quantified in the defined area 500µm apical to the root apex. Compared to CG, PG showed the following features: 15.13% less total bundle number; 17.8% less bundle size; significantly less connected fiber bundles; 9.2% more interrupted fiber bundles; and 3.85% more oriented toward the cervical in the bundle angulation. These results suggest that experimental periodontitis alters the PDL structure, leading to more interruptions of Sharpey's fiber attachments to the cementum.

Repeated botulinum treatment of rabbit masseter causes cumulative tissue damage.

OBJECTIVE: Botulinum neurotoxins (BoNT) are used in masticatory muscles for pain relief, unloading of the mandible, and cosmetic facial contouring. Treatment is often repeated every few months as function returns. This study assessed masticatory function and musculoskeletal structure after multiple BoNT treatment of the rabbit masseter. DESIGN: Female rabbits received 3 injections of BoNT (n = 13) or saline (n = 5) into one masseter muscle at intervals of 12 weeks. The contralateral side served as control. Periodic measurements of masticatory electromyography (EMG) and stimulated anterior bite force were made. After the final 12-week recovery interval, neuromuscular connection was investigated by stimulating the masseteric nerve to elicit an evoked EMG response. Mandibular specimens were collected for microCT analysis, and masseters were collected for histomorphometry and counts of replicating cells. RESULTS: Control and saline-injected muscles maintained consistent masticatory EMG and anterior bite force throughout the study. BoNT-injected masseters showed strong declines after each injection; during the 12-week recovery period, masticatory EMG and anterior bite force improved, although only electrical activity reached normal levels. Multiple injection resulted in persistently atrophied muscle fibers with fibrosis, and notable loss of bone from the mandibular body and condyle. The uninjected masseters of the BoNT group also showed evidence of mild toxin-related changes. CONCLUSIONS: Although muscle function is mostly regained after each injection, masseters receiving multiple doses of BoNT show extensive damage. In addition, mandibular bone density is decreased on the injected side.

Functional tooth mobility in young pigs.

Mobility is a fundamental characteristic of mammalian teeth, and has been widely used to determine individual tooth prognosis. However, the direction and extent of tooth movement under functional loads are unknown. This study investigated maxillary molar mobility, alveolar bending, and periodontal space (PDL) fluid pressure during mastication and masseter muscle contraction in young pigs, along with PDL space measurements. Twelve three-month-old farm pigs were instrumented with some or all of the following: (1) ultrasonic crystals, one implanted into the pulp chamber of a deciduous maxillary molar and additional crystals glued onto its buccal and palatal alveolar plates; (2) rosette strain gauges affixed to the buccal and palatal of alveolar ridges; (3) a pressure transducer inserted into palatal alveolar bone facing the PDL. Tooth mobility, alveolar bending, and fluid pressure were simultaneously recorded during unrestrained feeding and subsequent masseter muscle stimulation. The PDL widths were measured using micro-CT. The results indicate that during the power stroke of mastication, (1) the molar displaced buccally and apically (192 ± 95 µm) regardless of the side of chewing; (2) compressive bone strain was greater on the buccal than on the palatal alveolar plate; and (3) PDL pressure increased during the power strok (3.63 ± 0.80 kPa). Masseter contraction produced similar results but with generally lower values. The PDL widths were larger than the range of tooth mobility, and showed no correlation with the mobility. Thus occlusal function causes buccal tipping and intrusion of maxillary molars with concomitant compression of the buccal alveolar plate and raised pressure within the PDL space.

Alveolar bone loss and mineralization in the pig with experimental periodontal disease.

OBJECTIVE: To address how experimental periodontal disease affects alveolar bone mass and mineral apposition in a young pig model. MATERIALS AND METHODS: Seven three-month-old pigs were periodically inoculated with 4 types of periodontal bacteria, along with a ligature around the last maxillary deciduous molar for 8 weeks to induce periodontal disease (PG). Eight same-aged pigs served as the control (CG). Segmentations of 3D cone-beam CT images were performed to quantify volumes of the total alveolar bone, alveolar ridge, and all roots of the target molar. Calcein and alizarin were administered for labeling mineral apposition before euthanasia. The harvested molar blocks were sectioned and examined under epifluorescence. The inter-label distance between the two vital markers at regional bone surfaces were measured and mineral apposition rate (MAR) was calculated. RESULTS: A significant reduction of total alveolar bone volume was seen in PG with the major loss at the alveolar ridge. MAR was significantly higher at the root furcation region than those at both buccal and palatal ridges in CG. Compared with CG, PG animals showed more interrupted labeled bands with significantly lower MAR at the furcation region. MARs were positively associated with both the volumes of total alveolar bone and ridge in CG, but only with the total alveolar bone in PG. CONCLUSIONS: In young growing pigs, mineral apposition is region specific. The experimental periodontal disease not only leads to alveolar bone loss, but also perturbs mineral apposition for new bone formation, thus impairing the homeostasis of alveolar bone remodeling.

Botulinum toxin in masticatory muscles: short- and long-term effects on muscle, bone, and craniofacial function in adult rabbits.

Paralysis of the masticatory muscles using botulinum toxin (BTX) is a common treatment for cosmetic reduction of the masseters as well as for conditions involving muscle spasm and pain. The effects of this treatment on mastication have not been evaluated, and claims that the treatment unloads the jaw joint and mandible have not been validated. If BTX treatment does decrease mandibular loading, osteopenia might ensue as an adverse result. Rabbits received a single dose of BTX or saline into one randomly chosen masseter muscle and were followed for 4 or 12 weeks. Masticatory muscle activity was assessed weekly, and incisor bite force elicited by stimulation of each masseter was measured periodically. At the endpoint, strain gages were installed on the neck of the mandibular condyle and on the molar area of the mandible for in vivo bone strain recording during mastication and muscle stimulation. After termination, muscles were weighed and mandibular segments were scanned with micro CT. BTX paralysis of one masseter did not alter chewing side or rate, in part because of compensation by the medial pterygoid muscle. Masseter-induced bite force was dramatically decreased. Analysis of bone strain data suggested that at 4 weeks, the mandibular condyle of the BTX-injected side was underloaded, as were both sides of the molar area. Bone quantity and quality were severely decreased specifically at these underloaded locations, especially the injection-side condylar head. At 12 weeks, most functional parameters were near their pre-injection levels, but the injected masseter still exhibited atrophy and percent bone area was still low in the condylar head. In conclusion, although the performance of mastication was only minimally harmed by BTX paralysis of the masseter, the resulting underloading was sufficient to cause notable and persistent bone loss, particularly at the temporomandibular joint.