Tooth transplantation and replantation: Biological insights towards therapeutic improvements.

Transplantation and replantation of teeth are effective therapeutic approaches for tooth repositioning and avulsion, respectively. Transplantation involves transplanting an extracted tooth from the original site into another site, regenerating tissue including the periodontal ligament (PDL) and alveolar bone, around the transplanted tooth. Replantation places the avulsed tooth back to its original site, regenerating functional periodontal tissue. In clinical settings, transplantation and replantation result in favorable outcomes with regenerated PDL tissue in many cases. However, they often result in poor outcomes with two major complications: tooth ankylosis and root resorption. In tooth ankylosis, the root surface and alveolar bone are fused, reducing the PDL tissue between them. The root is subjected to remodeling processes and is partially replaced by bone. In severe cases, the resorbed root is completely replaced by bone tissue, which is called as "replacement resorption." Resorption is sometimes accompanied by infection-mediated inflammation. The molecular mechanisms of ankylosis and root resorption remain unclear, although some signaling mechanisms have been proposed. In this mini-review, we summarized the biological basis of repair mechanisms of tissues in transplantation and replantation and the pathogenesis of their healing failure. We also discussed possible therapeutic interventions to improve treatment success rates.

Platelet-derived growth factor-BB regenerates functional periodontal ligament in the tooth replantation.

Tooth ankylosis is a pathological condition of periodontal ligament (PDL) restoration after tooth replantation. Platelet-derived growth factor-BB (PDGF-BB) has been proposed as a promising factor for preventing tooth ankylosis. Using rat tooth replantation model, we investigated whether PDGF-BB accelerates the repair of PDL after tooth replantation without ankylosis, and its molecular mechanisms. In PDGF-BB pretreated replanted teeth (PDGF-BB group), ankylosis was markedly reduced and functionally organized PDL collagen fibers were restored; the mechanical strength of the healing PDL was restored to an average of 76% of that in non-replanted normal teeth at 21 days. The numbers of PDGF-Rß- and BrdU-positive cells in the periodontal tissues of the PDGF-BB group were greater than those of atelocollagen pretreated replanted teeth (AC group). Moreover, in the PDGF-BB group, the periodontal tissues had fewer osteocalcin-positive cells and decreased number of nuclear ß-catenin-positive cells compared to those in the AC group. In vitro analyses showed that PDGF-BB increased the proliferation and migration of human periodontal fibroblasts. PDGF-BB downregulated mRNA expressions of RUNX2 and ALP, and inhibited upregulatory effects of Wnt3a on ß-catenin, AXIN2, RUNX2, COL1A1, and ALP mRNA expressions. These findings indicate that in tooth replantation, topical PDGF-BB treatment enhances cell proliferation and migration, and inhibits canonical Wnt signaling activation in bone-tooth ankylosis, leading to occlusal loading of the PDL tissues and subsequent functional restoration of the healing PDL. This suggests a possible clinical application of PDGF-BB to reduce ankylosis after tooth replantation and promote proper regeneration of PDL.

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.

Transcriptome analysis of ankylosed primary molars with infraocclusion.

Primary molar ankylosis with infraocclusion can retard dental arch development and cause dental asymmetry. Despite its widespread prevalence, little is known about its molecular etiology and pathogenesis. To address this, RNA sequencing was used to generate transcriptomes of furcal bone from infraoccluded (n = 7) and non-infraoccluded (n = 9) primary second molars, all without succeeding biscuspids. Of the 18 529 expressed genes, 432 (2.3%) genes were differentially expressed between the two groups (false discovery rate < 0.05). Hierarchical clustering and principal component analysis showed clear separation in gene expression between infraoccluded and non-infraoccluded samples. Pathway analyses indicated that molar ankylosis is associated with the expression of genes consistent with the cellular inflammatory response and epithelial cell turnover. Independent validation using six expressed genes by immunohistochemical analysis demonstrated that the corresponding proteins are strongly expressed in the developing molar tooth germ, in particular the dental follicle and inner enamel epithelium. The descendants of these structures include the periodontal ligament, cementum, bone and epithelial rests of Malassez; tissues that are central to the ankylotic process. We therefore propose that ankylosis involves an increased inflammatory response associated with disruptions to the developmental remnants of the dental follicle and epithelial rests of Malassez.

Diagnosis of tooth ankylosis using panoramic views, cone beam computed tomography, and histological data: a retrospective observational case series study.

Objectives: The aim of this study was to determine whether cone beam computed tomography is a reliable radiological method to diagnose tooth ankylosis. Materials and methods: A series of teeth clinically diagnosed as ankylosed were collected after extraction in a private practice from 2009 to 2015 and analyzed retrospectively. Inclusion criteria comprised permanent molars extracted due to failed tooth eruption in the absence of any visible mechanical obstruction, existing panoramic view (PV), and cone beam computed tomography (CBCT) and histological sections of sufficient quality. The CBCT scans and PVs were evaluated twice for signs of ankylosis by two independent observers using the following score: clear signs, possible signs, and no signs. The histological sections were evaluated and graded similarly to the radiographs by a specialist blinded to the radiographs and treatment. Results: Out of an initial group of 22 patients, 9 subjects with 10 affected teeth were included for final evaluation. The age ranged from 8.3 to 17 years. No agreement was seen in comparing the PV scores to the histological sections. Fair to moderate agreement was seen in comparing the CBCT scores to the histological sections. All histologically confirmed ankylosis were detected in CBCT by both observers but some false positive results were found. Limitation: Only a small sample size was available as the disorder is rare. It is difficult to distinguish ankylosis from primary failure of eruption. Conclusion: CBCT images can be a useful adjunctive diagnostic tool to diagnose ankylosed teeth, but cannot be recommended as a single diagnostic modality as false positive results were found.

Introduction of a mixture of ß-tricalcium phosphate into a complex of bone marrow mesenchymal stem cells and type I collagen can augment the volume of alveolar bone without impairing cementum regeneration.

BACKGROUND: The purpose of this study is to evaluate whether ß-tricalcium phosphate (ß-TCP) could be a promising modality to help augment alveolar bone in periodontal tissue regeneration by bone marrow mesenchymal stem cells (BMMSCs). METHODS: Expanded BMMSCs and atelocollagen (Col) were mixed together (MSC/Col). A combination of ß-TCP with MSC/Col was also prepared (MSC/Col/TCP). MSC/Col/TCP or MSC/Col was transplanted into experimental periodontal Class III furcation defects that had been exposed to inflammation in beagle dogs. Periodontal tissue regeneration was evaluated by histologic and morphometric analyses at 4 and 8 weeks after transplantation. RESULTS: MSC/Col and MSC/Col/TCP enhanced periodontal tissue regeneration compared to Col and TCP/Col according to hematoxylin and eosin staining. The percentage of new cementum length in the MSC/Col/TCP group was not significantly different from that in the MSC/Col group at 4 and 8 weeks. On the other hand, the percentage of new bone area in the MSC/Col/TCP group was much higher than that in the MSC/TCP group at 4 weeks. However, at 8 weeks, no significant difference in new bone area was found between the two groups. In the MSC/Col/TCP group, ß-TCP was surrounded by newly formed bone. Multinucleated cells, which were positive for osteopontin and tartrate-resistant acid phosphatase, were present in the interconnected macropores of ß-TCP. CONCLUSION: These findings suggest that ß-TCP is applicable as a scaffold for BMMSCs transplantation and helps augment alveolar bone without impairing regeneration of cementum.

Delayed tooth replantation: MTA as root canal filling.

MTA has been investigated as a root-end filling material. Its mechanism of action has some similarities to that of Ca(OH)2. The purpose of this study was to evaluate the repair process taking place in the delayed replantation of monkey teeth using calcium hydroxide and MTA as root canal filling materials. Five monkeys had their lateral incisors extracted and bench-dried for 60 minutes. After root canal preparation, the teeth were assigned to two groups according to root canal filling material: I, calcium hydroxide; and II, MTA. The same treatment sequence was followed for both groups: coronal seal, periodontal ligament removal, immersion of the tooth in 2% acidulated-phosphate sodium fluoride, irrigation of the socket with saline and replantation. Both groups exhibited replacement resorption, areas of ankylosis and absence of inflammatory root resorption. Statistically similar results (p > 0.05) were observed for both groups regarding replacement root resorption, but the groups differed significantly (p < 0.05) regarding the occurrence of ankylosis. MTA may be a viable clinical option for filling teeth submitted to delayed replantation, and is an acceptable option for treating replanted permanent teeth in order to prevent tooth resorption, particularly when dressing changes are not possible.

Coronectomy of two neighbouring ankylosed mandibular teeth--a case report.

INTRODUCTION: Intentional partial odontectomy or coronectomy is primarily advocated in situations with intimate relationships between the roots of the tooth indicated for extraction and the inferior alveolar nerve (IAN). The aim of this report was to present a patient with two neighbouring infraoccluded teeth in the right mandible indicated for extraction prior to prosthetic rehabilitation, which were coronectomied as to prevent injuring of the IAN and causing iatrogenic fracture of the mandibular body. CASE REPORT: Coronectomy of both teeth was performed as recommended in the literature. The patient had no special discomfort after the operation or deficit in sensitive nerve function, and the wound healed uneventfully. The patient was followed regularly during a 2-year period. CONCLUSION: The presented case suggests coronectomy as quite beneficial solution for avoiding serious problems (injuring of the IAN and possible fracture of the mandible) and making feasible forthcoming prosthetic rehabilitation.

Effects of the bisphosphonate alendronate on molars of young rats after lateral luxation.

BACKGROUND AND AIM: The bisphosphonate alendronate (ALN) was employed with the aim of investigating its effects on dental and periodontal tissues after lateral luxation of developing molars. MATERIAL AND METHODS: Twenty-one-day-old Wistar rats had their second upper molars laterally luxated. Daily 2.5 mg kg(-1) ALN injections started at the day of the luxation; controls received sterile saline solution. The teeth were analyzed 7, 14, and 21 days after the procedure. On the days cited, the maxillae were fixed, decalcified, and embedded in paraffin or Spurr resin. The paraffin sections were stained with H&E, incubated for TRAP histochemistry or immunolabeled for osteopontin (OPN). Spurr ultrathin sections were examined in a transmission electron microscope. RESULTS: After 21 days, the root apex of luxated molars without ALN was wide open and disorganized and also covered by an irregular layer of cellular cementum, which was not observed in ALN-treated animals. Ankylosis sites were observed in ALN rats in both luxated and non-luxated teeth. The TRAP-positive osteoclasts were more numerous in ALN group, despite their latent ultrastructural appearance without the presence of resorption apparatus compared to controls. OPN immunolabeling revealed a thick immunopositive line in the dentin that must be resultant from the moment of the luxation, while ALN-treated specimens did not present alterations in dentin. CONCLUSION: The present findings indicate that alendronate inhibits some alterations in dentin and cementum formation induced by dental trauma.

Histologic and micro-computed tomographic analyses of replanted teeth stored in different kind of media.

INTRODUCTION: Coconut water (CW) and soy milk (SM) have been proposed as storage media for avulsed teeth because of their nutrients that preserve cell viability. The present study investigated the periodontal healing process of dog teeth replanted after storage in CW, SM, and whole milk (WM) using micro-computed tomographic (µCT) and histologic analyses compared with immediate tooth replantation. METHODS: Forty roots of 10 adult beagle dogs were extracted and subjected to the following protocols: immediate replantation after extraction (control), stored in CW with an adjusted pH, and SM and WM for 50 minutes before replantation. The animals were euthanized 28 days postoperatively, and the obtained specimens were scanned using a µCT scanner and subjected to routine processing for histometric analyses under an optical microscope. RESULTS: CW and SM performed similarly to WM; however, SM showed significantly higher ankylosis than the control group. CONCLUSIONS: Additionally, this study showed that the combined use of histologic analysis and µCT is a promising method to better identify tooth resorption and the repair process and to evaluate the total extension of the periodontium. CW as a storage medium is a promising transport media for avulsed teeth.