Compressed Periodontal Ligament Research Articles

GDF15 Contributes to the Regulation of the Mechanosensitive Responses of PdL Fibroblasts through the Modulation of IL-37.

During orthodontic tooth movement (OTM), areas of compressive and tensile forces are generated in the periodontal ligament (PdL), a mechanoreactive connective tissue between the teeth and alveolar bone. Mechanically stimulated PdL fibroblasts (PdLFs), the main cell type of PdL, express significantly increased levels of growth differentiation factor 15 (GDF15). In compressed PdL areas, GDF15 plays a fundamental role in modulating relevant OTM processes, including inflammation and osteoclast activation. However, the specific function of this factor in tensile areas has not yet been investigated. Thus, the aim of this study was to investigate the role of GDF15 in the mechanoresponse of human PdLFs (hPdLFs) that were exposed to biaxial tensile forces in vitro. Using siRNA-mediated knockdown experiments, we demonstrated that GDF15 had no impact on the anti-inflammatory force response of elongated hPdLFs. Although the anti-inflammatory markers IL1RN and IL10, as well as the activation of immune cells remained unaffected, we demonstrated an inhibitory role of GDF15 for the IL-37 expression. By analyzing osteogenic markers, including ALPL and RUNX2, along with an assessment of alkaline phosphatase activation, we further showed that the regulation of IL-37 by GDF15 modulates the osteogenic differentiation potential of hPdLFs. Despite bone resorption in tensile areas being rather limited, GDF15 was also found to positively modulate osteoclast activation in those areas, potentially by adjusting the IL-37 levels. In light of our new findings, we hypothesize that GDF15 modulates force-induced processes in tissue and bone remodeling through its various intra- and extracellular signaling pathways as well as interaction partners. Potentially acting as a master regulator, the modulation of GDF15 levels may hold relevance for clinical implications.

Differential gene expression and protein-protein interaction networks of human periodontal ligament stromal cells under mechanical tension

Orthodontic treatment is based on complex strategies and takes up to years until a desired therapeutic outcome is accomplished, implying long periods of high costs and discomfort for the patient. Choosing the optimal settings for force intensities in the initial phase of orthodontic tooth movement is the key to successful orthodontic treatment. It is known that orthodontic tooth movement is mainly mediated by tensile and compressive forces that are communicated to the alveolar bone via the periodontal ligament. While the revelation of the complex molecular network was already approached by transcriptomic analysis of compressed periodontal ligament cells, the entity of molecular key players activated by tensile forces remains elusive. Therefore, the aim of this study was to assess the effect of mechanical tensile forces on the gene expression profile of human primary periodontal ligament stromal cells, mimicking the initial phase of orthodontic tooth movement. A transcriptomic analysis of tension-treated and untreated periodontal ligament stromal cells yielded 543 upregulated and 793 downregulated differentially expressed genes. Finally, six highly significant genes were found in the transcriptome that are related to biological processes with relevance to orthodontic tooth movement, including apelin, fibroblast growth factor receptor 2, noggin, sulfatase 1, secreted frizzled-related protein 4 and stanniocalcin 1. Additionally, differences of gene expression profiles between individual cell donors showed a high effect size. Closer understanding of the roles of the identified candidates in the initial phase of orthodontic tooth movement could help to clarify the underlying mechanisms, which will be essential for the development of personalized treatment strategies in orthodontics.

Response of the tooth-periodontal ligament-bone complex to load: A microCT study of the minipig molar

One strategy evolved by teeth to avoid irreversible damage is to move and deform under the loads incurred during mastication. A key component in this regard is the periodontal ligament (PDL). The role of the bone underlying the PDL is less well defined. We study the interplay between the PDL and the underlying alveolar bone when loaded in the minipig. Using an Instron loading device we confirmed that the force-displacement curves of the molars and premolars of relatively fresh minipig intact mandibles are similar to those obtained for humans and other animals. We then used this information to obtain 3D images of the teeth before and after loading the tooth in a microCT such that the load applied is in the third linear part of the force displacement curve. We observed that at many locations there is a complimentary topography of the cementum and alveolar bone surface, strongly suggesting an active interplay between the tooth and the bone during mastication. We also observed that the loaded tooth does not come into direct contact with the underlying bone surface. A highly compressed layer of PDL is present between the tooth and the bone. The structure of the bone in the upper furcation region has a unique appearance with little obvious microstructure, abundant pores that have a large size range and at many locations the bone at the PDL interface has a needle-like shape. We conclude that there is a close interaction between the tooth, the PDL and the underlying alveolar bone during mastication. The highly compressed PDL layer that separates the tooth from the bone may fulfill a key shock absorbing function.

Effects of low magnitude high frequency mechanical vibration combined with compressive force on human periodontal ligament cells in vitro.

Vibration can be used to accelerate tooth movement, though the exact mechanisms remain unclear. This study aimed to investigate the effects of low magnitude high frequency (LMHF) vibration combined with compressive force on periodontal ligament (PDL) cells in vitro. Human PDL cells were isolated from extracted premolar teeth of four individuals. To determine the optimal frequency for later used in combination with compressive force, three cycles of low-magnitude (0.3 g) vibrations at various frequencies (30, 60, or 90 Hz) were applied to PDL cells for 20 min every 24 h. To investigate the effects of vibration combined with compressive force, PDL cells were subjected to three cycles of optimal vibration frequency (V) or 1.5 g/cm2 compressive force for 48 h (C) or vibration combined with compressive force (VC). Cell viability was assessed using MTT assay. PGE2, soluble RANKL (sRANKL), and OPG production were quantified by ELISA. RANKL, OPG, and Runx2 expression were determined using real-time PCR. Cell viability was decreased in groups C and VC. PGE2 and RANKL, but not OPG, were increased in groups V, C, and VC, thus increasing the RANKL/OPG ratio. The highest level was observed in group VC. sRANKL was increased in groups V, C, and VC; however, no significant different between the experimental groups. Runx2 expression was reduced in groups C and VC. Vibration increased PGE2, RANKL, and sRANKL, but not OPG and Runx2. Vibration had the additive effects on PGE2 and RANKL, but not sRANKL in compressed PDL cells.

Cementocyte cell death occurs in rat cellular cementum during orthodontic tooth movement.

To clarify the mechanism of root resorption during orthodontic treatment, we examined cementocyte cell death and root resorption in the cellular cementum on the pressure side during experimental tooth movement. Using 8-week-old male Wistar rats, the right first molar was pushed mesiobuccally with a force of 40 g by a Ni-Ti alloy wire while the contralateral first molar was used as a control. Localization and number of cleaved caspase-3-positive and single-stranded DNA (ssDNA) - positive cells were evaluated using dual-label immunohistochemistry with anticleaved caspase-3 and anti-ssDNA antibodies. In addition, tartrate-resistant acid phosphatase (TRAP)-positive cells in the cellular cementum were evaluated using TRAP histochemical staining. Caspase-3- and ssDNA-positive cells appeared at 12 hours, but were restricted to the compressed periodontal ligament (PDL) and not the cellular cementum. Cleaved caspase-3-positive cementocytes were observed in the cellular cementum adjacent to the compressed PDL on day 1. From days 2 to 4, the number of caspase-3- and ssDNA-positive cementocytes increased. TRAP-positive cells appeared on the cellular cementum at the periphery of the hyalinized tissue on day 7, and resorption progressed into the broad surface of the cementum by day 14. Cementocytes adjacent to the hyalinized tissue underwent apoptotic cell death during orthodontic tooth movement, which might have been associated with subsequent root resorption.

Expression of SOST/sclerostin in compressed periodontal ligament cells

Background/purposeBone resorption and inhibition of bone formation occur on the compressed side during orthodontic tooth movement. Bone formation inhibitory factors such as sclerostin (encoded by SOST) are secreted on the compressed side by periodontal ligament (PDL) cells. PDL cells control bone metabolism, and compressed PDL cells inhibit bone formation during orthodontic tooth movement. The aim of this study was to identify the inhibitory factors of bone formation in PDL cells. Materials and methodsChanges in SOST expression and subsequent protein release from human PDL (hPDL) cells were assessed using the real-time polymerase chain reaction (PCR), semiquantitative PCR, and immunofluorescence in hPDL cells subjected to centrifugal force (40g and 90g). To confirm the effects on bone formation, human alveolar bone-derived osteoblasts (hOBs) were grown with the addition of sclerostin peptide. In vivo, a compressive force was applied using the Waldo method in rats, and the distribution of sclerostin in PDL tissues was examined by immunohistochemistry. ResultsSOST expression was downregulated in vitro by centrifugation at 90g for 24 hours but upregulated by centrifugation at 40g based on real-time PCR, as was confirmed by immunofluorescence staining. The addition of sclerostin peptide significantly decreased the mineralized area in hOBs. However, slightly weakly sclerostin-positive PDL cells were observed on the compressed side in vivo. ConclusionThese results indicate that PDL cells subjected to light compressive force exhibit increased expression of SOST/sclerostin, which inhibits bone formation on the compressed side during orthodontic tooth movement.

Histochemical examination of cathepsin K, MMP1 and MMP2 in compressed periodontal ligament during orthodontic tooth movement in periostin deficient mice

The purpose of this study was to investigate immunolocalization of collagenolytic enzymes including cathepsin K, matrix metalloproteinase (MMP) 1 and 2 in the compressed periodontal ligament (PDL) during orthodontic tooth movement using a periostin deficient (Pn-/-) mouse model. Twelve-week-old male mice homozygous for the disrupted periostin gene and their wild type (WT) littermates were used in these experiments. The tooth movement was performed according to Waldo's method, in which elastic bands of 0.5mm thickness were inserted between the first and second upper molars of mice under anesthesia. At 1 and 3days after orthodontic force application, mice were fixed with transcardial perfusion of 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), and the first molars and peripheral alveolar bones were extracted for histochemical analyses. Compared with WT mice, immunolocalization of cathepsin K, MMP1 and MMP2 was significantly decreased at 1 and 3days after orthodontic tooth movement in the compressed PDL of Pn-/- mice, although MMP1-reactivity and MMP2-reactivity decreased at different amounts. Very little cathepsin K-immunoreactivity was observed in the assessed regions of Pn-/- mice, both before and after orthodontic force application. Furthermore, Pn-/- mice showed a much wider residual PDL than WT mice. Taken together, we concluded that periostin plays an essential role in the function of collagenolytic enzymes like cathepsin K, MMP1 and MMP2 in the compressed PDL after orthodontic force application.

Endotoxins potentiate COX-2 and RANKL expression in compressed PDL cells

This study aims to demonstrate in vitro the synergistic effect of orthodontic forces and periodontal pathogens on cyclooxygenase-2 regulation and the subsequent receptor activator of nuclear factor kappa-B ligand (RANKL) production from periodontal ligament (PDL) cells. In comparison to a control group, three experimental groups were formed from human primary PDL cells stressed with compressive forces, bacterial endotoxins, or a combination of both. Gene expression of cyclooxygenase-2 and RANKL was analysed with RT real-time PCR. The prostaglandin E2 production was determined with ELISA. A co-culture of PDL cells and an osteoclast-progenitor cell line was used in order to demonstrate the osteoclast formation effect caused by the simultaneous combined stress. The simultaneous combined stress resulted in a 56-fold up-regulation of cyclooxygenase-2 gene expression with a subsequent noticeable rise in the prostaglandin E2 in the culture medium. The RANKL/osteoprotegerin gene expression ratio was 50-fold up-regulated and the osteoclast formation assay revealed 153.5 ± 15.7 tartrate-resistant acid phosphatase (TRAP)-positive cells per well compared with 42.3 ± 3.8 TRAP-positive cells per well of the control group. The synergistic action of periodontal pathogens and orthodontic forces leads to an increased expression of cyclooxygenase-2 from PDL cells that intensify the RANKL production which in turn induces osteoclast differentiation and subsequent osteoclastogenesis. The present study puts an emphasis on the detrimental effect of orthodontic forces on patients with an active periodontal disease by underlining the significance of cyclooxygenase-2 activity and RANKL binding on the osteoclastogenesis process.

Experimental orthodontic tooth movement and extensive root resorption: periodontal and pulpal changes

Previous studies have reported changes both in dental pulp and in periodontal ligament (PDL) following orthodontic tooth movement. However, pulpal changes following extensive root resorption after orthodontic tooth movement have not been studied in detail. The aim of this study was therefore to evaluate inflammatory changes, both in the dental pulp and in the compressed PDL, after experimentally induced extensive root resorption. Extensive root resorption was induced in rats by the activation and re-activation of orthodontic force, with a short intervening period of no force application. The distribution of immune cells, nerve fibres and blood vessels was studied immunohistochemically using antibodies against CD68-immunoreactive (IR) cells, major histocompatibility complex (MHC) class II Ia-expressing cells, CD43-IR cells, protein gene product 9.5 (PGP 9.5), and laminin. In the compressed PDL of experimental first molars, significantly increased density of CD68-IR cells and MHC class II Ia-expressing cells were found, whereas the density of CD43-IR cells were unchanged when compared with control second molars. In the compressed PDL, there was an increased density of blood vessels, but no sprouting of nerve fibres. In the dental pulp, however, no increased density of immune cells or sprouting of nerve fibres was recorded. In conclusion, inflammation after extensive root resorption was confined to the compressed PDL, whereas the dental pulp was unaffected.

Effect of Ga‐Al‐As laser irradiation on COX‐2 and cPLA2‐α expression in compressed human periodontal ligament cells

Prostaglandin E(2) (PGE(2)), which has bone-resorptive activity, is produced by human periodontal ligament (PDL) cells in response to mechanical stress. We previously reported that low level laser (LLL) irradiation inhibited PGE(2) production in PDL cells in response to mechanical stress; however, the mechanism underlying this inhibitory effect was unclear. Thus, we sought to determine the mechanism underlying the inhibitory effect of LLL on PGE(2) production in compressed PDL cells. A compressive force of 2.0 g/cm(2) was applied for 24 hours to human PDL cells obtained from premolars extracted for orthodontic treatment. LLL irradiation (Ga-Al-As laser, 830 nm, 3.82 J/cm(2)) was applied 6 hours before, 1 hour before, and immediately after the application of compressive force. The mRNA expression of COX-2 and cPLA(2)-alpha was then examined by real-time PCR. LLL irradiation significantly inhibited COX-2 and cPLA(2)-alpha mRNA expression, which was increased in response to the application of a compressive force. Moreover, LLL irradiation immediately after compression had the strongest inhibitory effect on the expression of these genes. The inhibition of PGE(2) production by LLL irradiation in compressed PDL cells may be due to the inhibition of COX-2 and cPLA(2)-alpha expression and is most pronounced immediately after the application of a compressive force.

Early tooth movement with a clear plastic appliance in rats

In this preliminary study, we investigated early histologic changes of paradental tissues in response to a clear plastic appliance in rats. Fifteen rats were divided into 3 groups. Group I was the untreated controls; group II received a clear plastic appliance made from a model, with the maxillary left first molar repositioned mesially 0.5 mm from the origin; and group III had a closed-coil spring to move the molar mesially. Specimens were prepared in parasagittal sections, and changes in paradental tissues were evaluated on days 1, 4, and 7 by light microscopy. In group II, the periodontal ligament (PDL) was compressed in the bifurcation and apical areas of the roots of the molar. On day 7, the PDL of the apical and distal aspects of the roots and the bifurcation area showed further compression, with the PDL of the roots stretched along the mesial side. In group III, a disorganized and compressed PDL in the mesial cervical half and interradicular septum was observed, and the stretched fibers were at the distal aspects of the roots after days 4 to 7. Early histologic changes in response to the clear plastic appliance were intrusion and distal tipping despite the intended mesial movement. In this rat model, the observed histologic changes were subject to the direction and magnitude of forces generated by the clear aligner.

Periodontal response to experimental tooth movement by interrupted orthodontic force in rats

The upper first molars of Wistar rat were moved experimentally using an expansion screw in order to clarify whether an interrupted orthodontic force that maintains the blood circulation in compressed periodontal ligament (PDL) can cause efficient tooth movement without pathological damage. In the 30-μm group with moving distance of one-third of PDL width, degenerating tissue with toluidine blue staining and appreciable root resorption were not seen during the experimental period. On the other hand, in the 60-μm group with moving distance of two-third of PDL width degenerating tissue and severe root resorption appeared. On observation of bone formation using Villanueva fluorescent staining, a thick labeling line was observed on the tension side in both experimental groups. In statistical analysis about the ratio between mesial and distal PDL areas, expressing expansion rate of mesial PDL, there was no significant difference between 30-μm and control groups, except an experimental period of day 11. In contrast, significant differences were seen between 60-μm and control groups in 23–31 days after tooth movement. From these results, it is evident that the interrupted force that induces tooth movement within a distance of approximately one-third of PDL width enables tooth movement without toluidine blue-stained degenerating tissue, regarded as pathological damage, and promotes bone formation on the tension side.

Morphological changes in the rat periodontal ligament and its vascularity after experimental tooth movement using superelastic forces

The aim of this study was to statistically assess the morphological changes of the rat periodontal ligament (PDL) and its vascularity in relation to varied magnitudes of superelastic force in experimental tooth movement using nickel-titanium (NiTi) alloy wire. Forces of 0.8, 1.6, 4, 8, and 18 g were applied to the upper first molars of five groups of 10-week-old male Wistar rats (300-320 g) for 1, 7, 14, 21, and 28 days. A control group with no orthodontic appliance application was assessed in accordance with the five experimental periods. The specimens were observed under light microscopy, processed by computer imaging, and analysed statistically with Tukey's HSD non-parametric test. One day after the start of the experiment, a few blood vessels could be seen in the compressed PDL with forces of 0.8 and 1.6 g. The cross-sectional areas of blood vessels (CAV) and periodontal ligament (CAPL) in the experimental groups where a force of over 4 g was applied were significantly smaller than those where 0.8 and 1.6 g forces were used, and in the control group. On day 7, large CAV were seen in the 1.6, 4, and 8 g groups. On day 28, the 8 and 18 g groups showed significantly larger CAPL than the 0.8, 4 g, or control groups. The findings suggest that a light continuous force, under 1.6 g, maintains the vascular structure during experimental tooth movement. In contrast, a heavy continuous force over 4 g causes the vascular structure to be absent in the early stages of tooth movement, but a dynamic regeneration of the PDL with vascularity and expansion follows.

A new idea and method of tooth movement using a ratchet bracket

Since ideally effective tooth movement in orthodontics should occur without causing damage to the periodontal ligament (PDL), a new bracket with a ratchet-locking system, the 'Ratchet Bracket', was designed to produce tooth movement while maintaining blood circulation. To define the mechanism of the appliance, a histological study was carried out on four Beagle dogs (9 months old) and a clinical study on five female patients (11 years to 38 years 10 months of age). Five upper canines in the dogs were moved 1.82 mm per month. On light microscopic observations, vascular forms showed a round-oval shape, without undermining bone resorption. No root resorption was observed in the compressed PDL at days 1, 14, and 35 of the experimental period. On fluorescent images at day 46, distinctive bone formation was apparent at the tension side. In the clinical investigation, nine upper canines in the five female patients were moved 1.92 mm per month. A wide and long alveolar hard line was seen only on the tension side of the canines on dental radiographs, indicating bodily tooth movement, without obvious signs of root resorption in all subjects. Neither spontaneous pain nor pain during biting were reported. The findings indicate that use of the ratchet bracket could result in rapid and pain-free tooth movement with vascular clarity to maintain blood circulation in the PDL.

ALP activity decreased in compressed PDL cells obtained from severe orthodontically root resorption

Alkaline phosphatase (ALP) plays an important role in periodontal ligament (PDL) itself and alveolar bone in relation of calcification. However, very little is known about the relationship between external apical root resorption (EARR) during orthodontic treatment and ALP. We examined the effect of compressive force (CF) on ALP activity in human PDL (HPDL) cells from patients with severe orthodontically induced EARR. ALP activity, PGE 2 and IL-1β release were determined. The decrease of ALP and the increase of PGE 2 and IL-1β were greater in the severe root resorption (SRR) group than in the non-resorption (NR) group ( p < 0.001, two-way repeated measured ANOVA). NS-398, selective cyclooxygenase-2 (COX-2) inhibitor, and/or IL-1β antibody partially prevented the decrease in ALP activity. The presence of both factors together prevented almost completely the decrease in ALP activity to the control level ( p < 0.001). Therefore, the decrease in ALP activity was mediated mainly by PGE 2 and IL-1β produced by compressed PDL cells obtained from SRR. These results suggest that the decrease of ALP in compressed HPDL cells may be highly involved in the incidence of severe EARR during orthodontic treatment.

RANKL Increase in Compressed Periodontal Ligament Cells from Root Resorption

The ligand receptor activator of NFkappaB (RANKL) plays an important role in osteoclast formation. However, very little is known about the relationship between external apical root resorption during orthodontic treatment and RANKL. We hypothesized that compressive force is responsible for RANKL formation and up-regulation of osteoclastogenesis in periodontal ligament (PDL) cells from patients with severe orthodontically induced external apical root resorption. RANKL and osteoprotegerin (OPG) production, TRAP-positive cells, and resorptive pits were determined. The increase of RANKL and the decrease of OPG were greater in the severe root resorption group than in the non-resorption group. The numbers of TRAP-positive cells and resorptive pits were also increased in the severe root resorption group than in the non-resorption group. These results support the hypothesis that the compressed PDL cells obtained from tissues with severe external apical root resorption may produce a large amount of RANKL and up-regulate osteoclastogenesis.

Local OPG Gene Transfer to Periodontal Tissue Inhibits Orthodontic Tooth Movement

Previously, we discovered that RANKL expression is induced in compressed periodontal ligament cells, and that this promotes osteoclastogenesis on the compression side in orthodontic tooth movement. We hypothesized that local OPG gene transfer to the periodontium would neutralize the RANKL activity induced by mechanical compressive force, thereby inhibiting osteoclastogenesis and diminishing tooth movement. The upper first molars of six-week-old male Wistar rats were moved palatally by means of a fixed-orthodontic wire. A mouse OPG expression plasmid [pcDNA3.1(+)-mOPG] was constructed, and the production of functional OPG protein was confirmed in vitro. The inactivated HVJ envelope vector containing pcDNA3.1(+)-mOPG or PBS was injected periodically into the palatal periodontal tissue of upper first molars. When this local OPG gene transfer was performed, OPG production was induced, and osteoclastogenesis was inhibited. Local OPG gene transfer significantly diminished tooth movement. In this study, we report that OPG gene transfer to periodontal tissue inhibited RANKL-mediated osteoclastogenesis and inhibited experimental tooth movement.

Sympathectomy causes increased root resorption after orthodontic tooth movement in rats: immunohistochemical study.

Accumulating evidence suggests that the sympathetic nervous system modulates inflammatory responses and bone remodeling. We have studied the effects of sympathectomy and orthodontic tooth movement (OTM) on root resorption, immunocompetent cell recruitment, neuropeptide, neurokinin-1 receptor (NK1-R), and interleukin 6 (IL-6) expression. Experimental rats (n=8) had the right superior cervical ganglion surgically removed, whereas control rats (n=6) underwent sham surgery. Three days later, all rats had the right maxillary first molar moved mesially by an orthodontic appliance. The rats were perfused 13 days later, and the right maxillae were processed for immunohistochemistry by using primary antibodies directed against ED1 antigen, CD43, substance P (SP), NK1-R, neuropeptide Y (NPY), and IL-6. Following OTM, sympathectomized (SCGx) rats had significantly more root resorption (P<0.01) and SP-immunoreactive (IR) fibers (P=0.01) in the compressed periodontal ligament (PDL) compared with control rats. There was a significant decrease in recruitment of CD43+ cells in the pulp after OTM in SCGx rats compared with control rats (P=0.02). An upregulation of NK1-R immunoreactivity was observed surrounding the hyalinized tissue, and an increase in the number of NK1-R IR cells and density of SP-IR fibers was present in first molar pulp of all rats. NPY-IR fibers were absent in the compressed PDL of SCGx and control rats. Thus, OTM induces remodeling not only around the periodontal tissues, but also in the dental pulp. The sympathetic nerves have an inhibitory effect on hard tissue resorption and a stimulatory effect on CD43+ cell recruitment after OTM.

Apoptosis of periodontal ligament cells induced by mechanical stress during tooth movement.

The mechanical force generated during tooth movement creates compressed and cell-free areas in the periodontal membrane. The way in which periodontal ligament cells disappear at the compressed area during tooth movement remains unclear. In the present study we examined whether periodontal ligament cells undergo apoptosis by mechanical stress during tooth movement using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end-labelling method (TUNEL). TUNEL-positive stainings of periodontal ligament cells began to appear at the compressed areas 12 h after tooth movement, and the number of those cells reached maximum at 24 h after tooth movement. Thereafter TUNEL-positive cells disappeared at 48 h, and direct and undermining bone resorption began at the same area 72 h after tooth movement. These results showed that compressed periodontal ligament cells were eliminated by apoptosis in the early phase of tooth movement.

Cellular roles in relapse processes of experimentally-moved rat molars.

This histological study was designed to clarify the relapse processes of rat molars following orthodontic tooth movement by the Waldo method. An elastic band was inserted between the upper first and second molars, and removed either 7 or 21 days later. The relapse processes of first molars were examined at 1 or 4 days after completing the 7-day-insertion and at 5 or 10 days after the end of the 21-day-insertion. At the end of the 7-day-insertion period, the mean interdental space was 158 microns, but decreased rapidly, reaching 44 microns by 1 day after removal, and 16 microns by day 4. In the periodontal ligament (PDL) compressed by elastic band insertion, partial hyalinization and/or mineralization occurred. The stretched PDL consisted of irregularly-distributed, thin principal fibres, approximately 2.4 microns in diameter. On days 1 and 4 after removal, osteoclasts, macrophage-like cells and fibroblast-like cells were localized around hyalinized tissues, but thick principal fibres, approximately 3.8 microns in diameter, were regularly distributed throughout the PDL proper. The mean interdental space after the 21-day-insertion was 536 microns, but this had decreased to 108 microns by day 5, and 71 microns by day 10. On day 5 after removal, numerous osteoclasts were evenly distributed in the compressed PDL, while thick principal fibres, approximately 3.5 microns, were present in the stretched PDL. These results suggest that, during relapse of experimentally-moved rat molars, (i) rapid remodelling of the PDL and surrounding alveolar bones is the main cause of tooth relapse and (ii) hyalinized and/or mineralized tissues occurred by compression are rapidly resorbed by osteoclasts, macrophage-like cells and fibroblast-like cells.