A persistent mineralization process in alveolar bone throughout the postnatal growth stage in rats.

OBJECTIVE: Alveolar bone quality is essential for the maxillofacial integrity and function, and depends on alveolar bone mineralization. This study aims to investigate the in vivo changes in alveolar bone mineralization, from the perspective of mineral deposition and crystal transition in postnatal rats. DESIGN: Nine postnatal time points of Wistar rats, ranging from day 1 to 56, were set to obtain the maxillary alveolar bone samples. Each time point consisted of ninety rats, with 45 females and 45 males. Macromorphology of alveolar bone was reconducted by Micro-Computed Tomography and the mineral content was quantified via Thermogravimetric analysis, Scanning Electron Microscope, High-Resolution Transmission Electron Microscopy and vibrational spectroscopy. Furthermore, the crystallinity and composition were characterized by vibrational spectroscopy, X-ray Diffraction, X-ray Photoelectron Spectroscopy and Selected Area Electron Diffraction. RESULTS: The progressive increase of mineral deposition was accompanied by substantial growth in alveolar bone mass and volume in postnatal rats. Whereas the mineral percentage initially decreased and then increased, reaching a nadir on postnatal day 14 (P14) when tooth eruption was first observed. Besides, localized mineralization was initiated by the formation of amorphous precursors and then converted into mineral crystals, while there was no statistically significant change in the average crystallinity of the bone during growth. CONCLUSION: Mineralization of alveolar bone is ongoing throughout the early growth in postnatal rats. Mineral deposition increases with age, whereas the crystallinity remains stable within a certain range. Besides, the mineral percentage reaches its lowest point on P14, which may be attributed to tooth eruption.

Alveolar bone retention following treatment of dilacerated labial inversely impacted maxillary central incisors-a retrospective study.

The root of late-dental-age labial inversely impacted maxillary central incisors (LIIMCIs) typically develops to severe dilacerated morphology. Therefore, reliable posttreatment periodontal estimates of orthodontic treatment prognosis would be critical to the treatment value of impacted incisors. This study aims to analyze further changes in dimensions of the alveolar bone following the closed-eruption treatment of late-dental-age dilacerated LIIMCIs. Cone beam computed tomography (CBCT) scanning data of 16 patients with unilateral dilacerated late-dental-age LIIMCIs were collected, including the pretreatment (T1) and at the 2.23 ± 0.78 years follow-up stage (T2) respectively. Patients underwent closed-eruption treatments to bring the impacted incisor into the dental arch. Dolphin imaging software was used to measure alveolar bone height labially, palatally, and proximally to the site at T1 and T2, as well as alveolar bone thicknesses at 0, 2, 4, 6 and 8 mm below the initial measurement plane (IMP). The alveolar bone heights on the impacted and contralateral sides increased from T1 to T2 (p < 0.05). Alveolar bone growth on both sides had no significant difference. In T2, the mean values of labial and distal alveolar heights on the contralateral sides were greater than on the impacted sides (p < 0.05). The mean values of total alveolar bone thicknesses on the impacted sides in T1 were significantly smaller than those on the contralateral sides in IMP-0, 2, 4, 6, 8 (p < 0.05). The total thicknesses on the impacted sides in T2 increased and were significantly greater than on the contralateral sides (p < 0.05), except for the thickness in IMP-0. The closed-eruption treatment of dilacerated late-dental-age LIIMCIs results in no significant changes to alveolar bone height, except on the labial and distal sides, with increased alveolar bone thickness, suggesting that this approach may be viable first choice therapy for non-extraction orthodontic cases.

Application of cryopreservation to tooth germ transplantation for root development and tooth eruption.

We cryopreserved mouse tooth germs with widely open cervical margins of the enamel organ to overcome difficulties in cryoprotectant permeation and tested their efficacy by transplanting them into recipient mice. The upper right first molar germs of 8-day-old donor mice were extracted and categorized into the following four groups according to cryopreservation time: no cryopreservation, 1 week, 1 month, and 3 months. The donor tooth germs were transplanted into the upper right first molar germ sockets of the 8-day-old recipient mice. The upper left first molars of the recipient mice were used as controls. The outcome of the transplantation was assessed at 1, 2, and 3 weeks after transplantation. Stereomicroscopic evaluation revealed that most of the transplanted teeth erupted by 3 weeks after transplantation. Micro-computed tomography analysis revealed root elongation in the transplanted groups as well as in the controls. There was no significant difference between the cryopreserved and non-cryopreserved transplanted teeth, but the roots of the cryopreserved teeth were significantly shorter than those of the control teeth. Histological examination revealed root and periodontal ligament formations in all the transplanted groups. These results suggest that the transplantation of cryopreserved tooth germs facilitates subsequent root elongation and tooth eruption.

The Dual Effects of Reactive Oxygen Species on the Mandibular Alveolar Bone Formation in SOD1 Knockout Mice: Promotion or Inhibition.

The status of reactive oxygen species (ROS) correlates closely with the normal development of the oral and maxillofacial tissues. Oxidative stress caused by ROS accumulation not only affects the development of enamel and dentin but also causes pathological changes in periodontal tissues (periodontal ligament and alveolar bone) that surround the root of the tooth. Although previous studies have shown that ROS accumulation plays a pathologic role in some oral and maxillofacial tissues, the effects of ROS on alveolar bone development remain unclear. In this study, we focused on mandibular alveolar bone development of mice deficient in superoxide dismutase1 (SOD1). Analyses were performed using microcomputerized tomography (micro-CT), TRAP staining, immunohistochemical (IHC) staining, and enzyme-linked immunosorbent assay (ELISA). We found for the first time that slightly higher ROS in mandibular alveolar bone of SOD1(-/-) mice at early ages (2-4 months) caused a distinct enlargement in bone size and increased bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and expression of alkaline phosphatase (ALP), Runt-related transcription factor 2 (Runx2), and osteopontin (OPN). With ROS accumulation to oxidative stress level, increased trabecular bone separation (Tb.Sp) and decreased expression of ALP, Runx2, and OPN were found in SOD1(-/-) mice at 6 months. Additionally, dosing with N-acetylcysteine (NAC) effectively mitigated bone loss and normalized expression of ALP, Runx2, and OPN. These results indicate that redox imbalance caused by SOD1 deficiency has dual effects (promotion or inhibition) on mandibular alveolar bone development, which is closely related to the concentration of ROS and the stage of growth. We present a valuable model here for investigating the effects of ROS on mandibular alveolar bone formation and highlight important roles of ROS in regulating tissue development and pathological states, illustrating the complexity of the redox signal.

A study on the morphologic change of palatal alveolar bone shape after intrusion and retraction of maxillary incisors.

The purpose of this study is to evaluate the changes in the palatal alveolar bone thickness and find the factors related to the resorption of the palatal alveolar bone caused by tooth movement after the maxillary incisors were retracted and intruded during orthodontic treatment. The study group comprised of 33 skeletal Class II malocclusion patients who underwent extraction for orthodontic treatment. Palatal alveolar bone thickness changes and resorption factors were identified and analyzed. The changes of maxillary central incisors and palatal alveolar bone thickness were measured, and the corresponding sample t test was performed using SPSS (IBM SPSS version 22). The amount of palatal alveolar bone resorption was measured and various parameters were analyzed to determine which factors affected it. Correlation analysis adopting the amount of palatal alveolar bone resorption as a dependent variable demonstrated that the SNB, mandibular plane angle, and the inclination of the maxillary central incisor were significantly correlated with before treatment. On the other hand, mandibular plane angle, angle of convexity, the inclination of the upper incisor, and the occlusal plane (UOP, POP) were significantly correlated with post-treatment. In addition, the variables related to palatal contour (PP to PAS, SN to PAS, palatal surface angle) and occlusal planes (UOP/POP) were significantly correlated with the difference in palatal bone resorption. During initial diagnosis, high angle class II with normal upper incisor inclination can be signs of high-risk factors. In addition, maintaining the occlusal plane during treatment helps to prevent palatal bone loss.

Dual role of phosphatidylserine and its receptors in osteoclastogenesis.

Fusion and apoptosis share a breakdown of the membrane phospholipids asymmetry, modes of which are largely unknown in osteoclastogenesis. Here, we investigated the externalization of phosphatidylserine (PS) and its receptors, and their biological functions in osteoclastogenesis. Strong immunoreactivities in vivo for the PS receptors TIM4, BAI1, and STAB2 were observed in the TRAP-positive multinucleated cells in the alveolar bone that was being remodeled around the developing dental follicles in rats. These receptors were significantly upregulated during M-CSF/RANKL-induced in vitro osteoclastogenesis using mouse bone marrow-derived cells. PS externalization in preosteoclasts was increased by the M-CSF/RANKL treatment. Multinucleation of preosteoclasts was markedly inhibited by antibodies against PS and its receptors. Among the investigated lipid transporter proteins, floppases (Abcb4, Abcc5, and Abcg1) were upregulated, whereas flippases (Atp11c and Atp8a1) downregulated during osteoclastogenesis. Preosteoclast fusion was markedly blocked by the ATPase inhibitor Na3VO4 and siRNAs against Abcc5 and Abcg1, revealing the importance of these lipid transporters in PS externalization. Further, the levels of Cd47 and Cd31, don't-eat-me signal inducers, were increased or sustained in the early phase of osteoclastogenesis, whereas those of AnnexinI and Mfg-e8, eat-me signals inducers, were increased in the late apoptotic phase. In addition, Z-VAD-FMK, a pan caspase inhibitor, had no effect on preosteoclast fusion in the early phase of osteoclastogenesis, whereas Abs against PS, TIM4, and BAI1 decreased osteoclast apoptosis during the late phase. These results suggest that PS externalization is essential for the whole process of osteoclastogenesis and share PS receptors and transporters in the early stage fusion and late stage apoptosis. Therefore, modulation of PS and its receptors could be a useful strategy to develop anti-bone resorptive agents.

Comparison of morphological quantitative characteristics of the newly formed alveolar bone after the application of demineralized bone matrix and cortical bone allografts / Comparación de las características morfológicas cuantitativas del hueso alveolar recién formado después de la aplicación de matriz ósea desmineralizada y aloinjertos de hueso cortical

The allografts were used to obtain sufficient alveolar bone tissue for proper dental implant placement. The objective of the present study was to evaluate the morphological and quantitative characteristics (cellular and collagen densities) of the newly formed alveolar bone with the application of cortical bone (CB) and demineralized bone matrix (DBM) allografts. Six samples of alveolar bone tissue from 5 patients (50 ± 6.3 years) were obtained after 6 months of application of the allografts and immediately before the placement of the dental implants. The samples were fixed (buffered formaldehyde, pH7.2), decalcified (EDTA 10 %) and histologically processed (HE and Picro-Sirius) for histologic analysis. Morphological analysis revealed presence of osteocytes and trabeculae in neoformed bone tissue near the allografts and absence of inflammatory and allergic cells; the remnants of CB were located mainly in the periphery of the bone tissue and the remnants of DBM were more incorporated into the tissue. Osteogenitor cells were observed around the remaining material. The cell density was not modified in newly formed bone tissue with the application of both allografts as compared to mature bone tissue. The density of the type I and III collagens present in the osteoids interspersed with the remainder of the materials showed a tendency to increase in the samples treated with DBM. It was concluded that by the histological characteristics observed both grafts were biocompatible, however the bone treated with DBM presented better incorporation and a tendency of increase of the collagen content in the remnant region of the allografts.

Los aloinjertos son utilizados para obtener tejido óseo alveolar apropiado para la colocación correcta del implante dental. El objetivo de este trabajo fue evaluar las características morfológicas y cuantitativas (densidades celulares y de colágeno) del hueso alveolar recién formado con la aplicación de aloinjertos de hueso cortical (CB) y matriz desmineralizada de hueso (DBM). Seis muestras de tejido óseo alveolar fueron obtenidas de 5 pacientes (50 ± 6,3 años) después de 6 meses de aplicación de los aloinjertos e inmediatamente antes de la colocación de los implantes dentales. Las muestras fueron fijadas (formaldehído tamponado, pH 7,2), descalcificadas (EDTA al 10%) y procesadas histológicamente (HE y Picro-Sirius) para el análisis histológico. El análisis morfológico reveló la presencia de osteocitos y trabéculas en el tejido óseo neoformado cerca de los aloinjertos y la ausencia de células inflamatorias y alérgicas; los remanentes de CB se ubicaron principalmente en la periferia del tejido óseo y los remanentes de DBM se incorporaron más en el tejido. Se observaron células osteogenitoras alrededor del material restante. La densidad celular no se modificó en el tejido óseo recién formado con la aplicación de ambos aloinjertos en comparación con el tejido óseo maduro. La densidad de los colágenos de tipo I y III presentes en los osteoides intercalados con el resto de los materiales mostró una tendencia a aumentar en las muestras tratadas con DBM. Se concluyó que, debido a las características histológicas observadas, ambos injertos fueron biocompatibles, sin embargo, el hueso tratado con DBM presentó una mejor incorporación y una tendencia al aumento del contenido de colágeno en la región remanente de los aloinjertos.

Growth-dependent phenotype in FasL-deficient mandibular/alveolar bone.

FASL (CD178) is known for its role in triggering apoptosis, mostly in relation with immune cells but additional functions have been reported more recently, including those in bone development. Examination of postnatal FasL-deficient mice (gld) showed an increased bone deposition in adult mice when compared with wild types. However, a different phenotype was observed prenatally, when the gld bone was underdeveloped. The aim of the following investigation was to evaluate this indication for an growth-dependent bone phenotype of gld mice and to search for the 'switch point'. This study focused on the mandibular/alveolar bone as an important structure for tooth anchorage. In vivo micro-computed tomography (CT) analysis was performed at different stages during the first month (6, 12 and 24 days) of postnatal bone development. In 6-day-old gld mice, a decrease in bone volume/tissue volume (BV/TV), trabecular thickness and trabecular number was revealed. In contrast, the 12-day-old gld mice showed an increased BV/TV and trabecular thickness in the alveolar bone. The same observation applied for bone status in 24-day-old gld mice. Therefore, changes in the bone phenotype occurred between day 6 and 12 of the postnatal development. The switch point is likely related to the changing proportion of bone cells at these stages of development, when the number of osteocytes increases. Indeed, the immunohistochemical analysis of FASL localized this protein in osteoblasts, whereas osteocytes were mostly negative at examined stages. The impact of FASL particularly on osteoblasts would agree with an earlier in vivo observed effect of FASL deficiency on expression of Mmp2, typical for osteoblasts, in the gld mandibular/alveolar bone. Notably, an age-dependent bone phenotype was reported in Mmp2-deficient mice.

New Bone Formation Using an Extracted Tooth as a Biomaterial: A Case Report with Histologic Evidence.

This case report aims to demonstrate the regenerative potential of particles obtained from a crushed extracted tooth. Following tooth removal, the clean root was ground and the dentin and cementum granules were grafted into a fresh extraction socket for a ridge preservation procedure. After 24 weeks, a successful implant placement was allowed. Tissue healing was evaluated by histologic and radiologic analysis. The volume of the ridge was preserved. Histologically, a dentin-bone complex was reported. New bone formation was evident, with an intimate contact between bone and both dentin/cementum. This novel procedure suggests the use of tooth particles as graft material.

Numerical investigations of bone remodelling around the mouse mandibular molar primordia.

The formation of the alveolar bone, which houses the dental primordia, and later the roots of tooth, may serve as a model to approach general questions of alveolar bone formation. In this respect, this study aimed to investigate the potential interactions between the alveolar bone formation and tooth eruption by using finite element (FE) methods, and to figure out whether the expanding tooth systems induce shear stresses that lead to alveolar bone formation. 3D geometric surface models were generated from the 3D histological data of the heads of mice (C57 Bl/6J) ranging from stages embryonic (E) to postnatal (P) stages E15 to P20 using the reconstruction software 3-Matic. Bone, dentin, enamel and dental follicle around the primordia were generated and converted into 3D FE models. Models were imported into the FE software package MSC.Marc/Mentat. As material parameters of embryonic dentine, pulp, enamel, dental follicle, and bony structures basically are unknown, these were varied from 1% to 100% of the corresponding known material parameters for humans and a sensitivity analysis was performed. Surface loads were applied to the outside surface of dental follicle ranging from 0.1 to 5.0N/mm2. The validity of the model was analysed by comparing the activity pattern of the alveolar bone as determined in the histological study with the loading pattern from the numerical analysis. The results show that when varying the surface loads, the distribution of shear stresses remained same, and while varying the material properties of the hard tissues, the location of highest shear stresses remained stable. Comparison of the histologically determined growth regions with the distribution of shear stresses computed in the numerical model showed a very close agreement. The results provide a strong proof to support Blechschmidt's hypothesis that the bone in general is created under the influence of shear forces.