Analysis of the cells isolated from epithelial cell rests of Malassez through single-cell limiting dilution.

The epithelial cell rests of Malassez (ERM) are essential in preventing ankylosis between the alveolar bone and the tooth (dentoalveolar ankylosis). Despite extensive research, the mechanism by which ERM cells suppress ankylosis remains uncertain; perhaps its varied population is to reason. Therefore, in this study, eighteen unique clones of ERM (CRUDE) were isolated using the single-cell limiting dilution and designated as ERM 1-18. qRT-PCR, ELISA, and western blot analyses revealed that ERM-2 and -3 had the highest and lowest amelogenin expression, respectively. Mineralization of human periodontal ligament fibroblasts (HPDLF) was reduced in vitro co-culture with CRUDE ERM, ERM-2, and -3 cells, but recovered when an anti-amelogenin antibody was introduced. Transplanted rat molars grown in ERM-2 cell supernatants produced substantially less bone than those cultured in other cell supernatants; inhibition was rescued when an anti-amelogenin antibody was added to the supernatants. Anti-Osterix antibody staining was used to confirm the development of new bones. In addition, next-generation sequencing (NGS) data were analysed to discover genes related to the distinct roles of CRUDE ERM, ERM-2, and ERM-3. According to this study, amelogenin produced by ERM cells helps to prevent dentoalveolar ankylosis and maintain periodontal ligament (PDL) space, depending on their clonal diversity.

Aberrantly elevated Wnt signaling is responsible for cementum overgrowth and dental ankylosis.

Vertebrate teeth are attached to the jawbones using a variety of methods but in mammals, a fibrous connection is the norm. This fibrous periodontal ligament (PDL) allows teeth to move in the jawbones in response to natural eruptive forces, mastication, and orthodontic tooth movement. In some disease states the PDL either calcifies or is replaced by a mineralized tissue and the result is ankylosis, where the tooth is fused to the alveolar bone. To understand how the PDL maintains this fibrous state, we examined a strain of mice in which tooth movement is arrested. DaßcatOt mice express a stabilized form of ß-catenin in DMP1-positive alveolar bone osteocytes and cementocytes, which results in elevated Wnt signaling throughout the periodontium. As a consequence, there is an accrual of massive amounts of cellular cementum and alveolar bone, the PDL itself calcifies and teeth become ankylosed. These data suggest that to maintain its fibrous nature, Wnt signaling must normally be repressed in the PDL space.

A qualitative investigation of RANKL, RANK and OPG in a rat model of transient ankylosis.

INTRODUCTION: Previous studies have found ankylosis occurs as a part of the inflammatory process of aseptic root resorption initiated in a rat model. The physiologic mechanisms behind the development of dentoalveolar ankylosis and healing response are still unclear. While receptor activator of nuclear factor-κß ligand (RANKL), receptor activator of nuclear factor-κß (RANK) and osteoprotegerin (OPG) have gained momentum in the understanding of resorption, no study to date has investigated their role in dentoalveolar ankylosis. AIMS: The aims of this study were to investigate if, and when, ankylosis occurred in the rat PDL, whether the resolution of ankylosis occurred with time and, finally, to observe the expression of RANKL, RANK and OPG during the ankylotic process. MATERIALS AND METHODS: Dry ice was applied for 20 minutes to the upper right first molar crown of 15 eight-week-old, male Sprague-Dawley rats. An additional three rats served as untreated external controls. Groups of three rats were sacrificed after the thermal insult on day 0, 4, 7, 14 and 28 respectively. Each maxilla was dissected out and processed for histological examination and RANKL, OPG and RANK immunohistochemistry. RESULTS: By the use of light microscopy and H&E staining, no ankylosis was detected in the external control group and the experimental groups at days 0 and 4. On day 7, disruption within the periodontal ligament was observed in the interradicular region and the initial signs of ankylosis were seen in the form of finger-like projections extending from the alveolar bone towards the cementum. Fourteen days after the thermal insult, all animals exhibited extensive ankylosis that spanned the entire interradicular periodontal space. At 28 days, the development of ankylosis appeared to have ceased and repair was observed, together with an intact periodontal ligament in all but one rat. Positive staining results were obtained with RANKL, RANK and OPG antibodies. The expressions of RANKL, RANK and OPG were similar in the external control group, 0-, 4-, and 28-day experimental groups. In the 7- and 14-day experimental groups, RANKL, RANK and OPG were expressed in the blood vessels within the ankylotic regions. CONCLUSIONS: During the development of ankylosis and its resolution, it was concluded from their simultaneous presence that there is a complex interaction between RANKL, RANK and OPG that requires further investigation.

A role for TNF in bone resorption of deciduous molars in human beings.

In this study, tumor necrosis-alpha was sampled from the gingival crevice of human deciduous molars; this was compared with values measured from the crevice of those deciduous molars missing a permanent successor, and from the crevice of deciduous ankylosed molars. Tumor necrosis-alpha was harvested from the gingival crevice with magnetic microspheres coated with tumor necrosis-alpha antibodies. The amount of bead-bound tumor necrosis-alpha was quantified with the use of an enzyme-linked immunosorbent assay. One hundred seven sites (from 41 patients) were sampled; for each patient, the normal value was compared with either the molars missing a permanent successor or ankylosed value. The tumor necrosis-alpha levels were 1.6 times higher from the crevice of ankylosed deciduous molars when compared with normal deciduous molars and 2.6 times higher from the crevice of sites with a molar missing a permanent successor. The mean and standard error mean distribution of tumor necrosis-alpha expressed as picograms was: normal molars 91 pg (standard error mean +/- 20), ankylosed molars 150 pg (standard error mean +/- 31), and missing permanent successor 236 pg (standard error mean +/- 67). Analysis of variance showed the difference among the 3 means was close to attaining significant difference (F [2.104] = 2.7905, P =.066). Multiple comparison procedures indicated that the mean for molars missing a permanent successor and the normal groups were significantly different, P =.05. The results of this study suggest tumor necrosis-alpha values are elevated in the gingival crevice of deciduous molars with ankylosis and where the permanent tooth bud is congenitally missing.