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Interleukin-1beta-induced release of matrix proteins into culture media causes inhibition of mineralization of nodules formed by periodontal ligament cells in vitro.

The mechanism by which interleukin-1beta (IL-1) inhibits the formation of mineralized tissue nodules by periodontal ligament (PDL) cells in vitro was investigated through the processes of morphological analysis, immunoprecipitation, and Northern blot analysis. PDL cells were obtained from a 2-day-old coagulum in tooth socket and cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bone serum (FBS) and antibiotics. Confluent cells were grown for up to 3 weeks in the presence of ascorbic acid (AA), beta-glycerophosphate (GP), and dexamethasone (Dex), or IL-1. PDL cells cultured in the presence of GP and AA did not differentiate, but those treated with Dex, GP, and AA (Dex group) underwent differentiation, showing four stages (confluent, multilayer, nodule, and mineralization) of disparate morphological characteristics. In contrast, the cells treated with IL-1, Dex, GP, and AA (IL-1 group) did form multilayers but failed to form mineralized nodules. Electron microscopy demonstrated that the Dex-induced mineralized nodules contain multilayers of fibroblastic cells, numerous collagen fibrils, and dense globular as well as fused electron dense patches that are associated with numerous apatite crystals. The nodule-like structures in the IL-1 group were also comprised of multilayered fibroblastic cells, but they contained only a small number of collagen fibrils, and no dense globular or fused patches. Von Kossa staining confirmed the presence of numerous mineralized nodules in the Dex group and their scarceness in the IL-1 group. Northern blot analysis of IL-1-treated cells, however, revealed the presence of mRNAs for type I collagen (Col I), secreted protein, acidic and rich in cysteine (SPARC), osteopontin (OPN), alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OC), whose expression patterns and levels were comparable to those of the Dex group. Immunoprecipitation analysis of OPN and BSP in the cell/matrix layers and the culture media after [35S]-methionine labeling showed their deposition primarily in the mineralized nodules of the Dex group, and their release into the media in the IL-1 group. Immunogold labeling demonstrated the location of OPN and BSP in mineralized nodules of the Dex group, but no significant labeling occurred in the nodule-like structures from the IL-1 group. Interestingly, IL-1 treatment increased the expression of collagenase mRNA by sevenfold, compared with that of the Dex group. These data suggest that the IL-1-induced formation of unmineralized nodules by PDL cells results not so much from the downregulated formation of matrix proteins, which plays a crucial role in the mineralization process, as from their release into the culture media. Finally, collagenase synthesis upregulated by IL-1 may be involved in this process.

Animals↗

[Osteonecrosis of the jaws by long term therapy with bisphosphonates].

For several decades bisphosphonates have been used to reduce skeletal related events in patients with both osteoporosis or bone metastases. Under long term application, besides the known therapy side effects, a new clinical picture has been described within the last few years. This is osteonecrosis of the jaws, which is characterized by its difficulty in treatment. Besides exposed jaw bone, the start of the disease usually lacks any symptoms. The typical clinical symptoms then are foetor ex ore, swelling, exsudation, loosening of teeth, pain or paresthesia. Later oro-antral/nasal or oro-cutaneous fistula can develop. The X-ray shows persisting tooth sockets after extractions and later cloudy radio-lucency, sequestra or fractures. The patient exposed to bisphosphonate can be grouped according to the risk for osteonecrosis: high risk patients with intravenous bisphosphonate therapy and additional chemo-, radiation or corticoid therapy--predominantly patients with a malignant underlying disease and bone metastases low risk patients with an oral bisphosphonate therapy without additional chemo-, radiation or corticoid therapy--preferably patients with non-corticoid-induced osteoporosis. Before starting a bisphosphonate therapy possible causes of infection should be treated and risk of injuries to the mucosa should be reduced according to the individual risk profile. This is supplemented by information of the patient about the risk of necrosis and the possibilities for prevention. Regular dental recall under bisphophonate therapy is emphasised for early recognition of possible problems. Prophylaxis is recommended for the prevention of periodontal infection combined with a follow up of removable denture for possible ulcera. Generally, conservative treatment measures are preferred to surgical ones. Inevitable operations are carried out non-traumatically using broad spectrum antibiotic prophylaxis until the day of suture removal (not before day 10). Long term follow up examinations are recommended.Patients with dental implants inserted before a bisphophonate therapy should be subject to intensive recall examinations. For patients undergoing or following a bisphosphonate therapy the indication for dental implants should be as strict as for patients following head and neck radiation therapy. In the present for patients with osteonecrosis, even after healing, dental implants are regarded as contra-indication. Therapy of the necrosis often requires general anaesthesia, hospitalisation, naso-gastral feeding tube and intravenous, systemic antiinfective treatment. The necrosis is removed completely and a tension free wound closure with vascularised tissue is intended. A literature review shows the metabolic effect of biphosphonates, the known pathogenesis of the bisphosphonate-induced jaw necrosis. It is essential to develop interdisciplinary communication, aiming at a joint care for this group of concerned patients and involving not only those medical disciplines, which order and use bisphosphonates, but especially dentists and maxillofacial surgeons.

Alveolar Process↗

Oral radiographs in the detection of early signs of osteoporosis.

A study was conducted to determine whether radiographic changes could be detected in the mandible of patients with mild-to-moderate postmenopausal osteoporosis and whether these changes could be used as a diagnostic tool to differentiate normal from osteoporotic patients. Subjects were classified as either osteoporotic (21) or normal (14) on the basis of bone density measurements of the lumbar spine and femoral neck, as determined by dual-photon absorptiometry. Mandibular bone density measurements were made on panoramic and periapical radiographs and expressed in terms of millimeters of aluminum equivalent. Thickness of the cortex at the angle of the mandible, sinus floor, and lamina dura of the tooth socket was also measured. There were no significant differences in any of the mandibular measurements between the normal and osteoporotic subjects. Whereas the skeletal bone measurements were correlated with each other, there was no correlation between skeletal and mandibular bone measurements. Women with mild-to-moderate osteoporosis could not be distinguished from women with normal bone density with the method described in this article.

Bone Density↗

Symptomatic foreign body reaction to haemostatic alginate.

Kaltostat, a haemostatic wound dressing made from non-woven sodium calcium alginate fibres is becoming more frequently and widely used by practitioners. We report a florid foreign body giant cell reaction elicited by Kaltostat which had been used to obtain haemostasis in an apicectomy cavity on an upper lateral incisor approximately 7 months earlier. The case demonstrates that alginate fibres left in situ may elicit a long-lasting and symptomatic adverse foreign body reaction. This material should be reserved for problematic haemorrhage and be removed from the tooth socket soon after haemostasis.

Alginates↗

Three-dimensional localization of maxillary canines with cone-beam computed tomography.

INTRODUCTION: Precise 3-dimensional (3D) localization of impacted canines is central to their clinical management. Recently introduced dental 3D volumetric imaging systems make precise localization possible. The purpose of this study was to describe the spatial relationship of impacted canines by using images obtained with the NewTom QR-DVT 9000 (QR Srl, Verona, Italy). METHODS: Unilaterally and bilaterally impacted canines (n = 27) from 19 consecutive patients (15 female, 4 male) were evaluated on images taken with the NewTom QR-DVT 9000. The spatial relationships of the impacted canines relative to adjacent structures and incisor resorption were assessed with 3D visualization software. RESULTS: Most (92.6%) of the 27 impactions were palatal. Incisor resorption adjacent to the impacted canine was present in 66.7% of the lateral incisors and 11.1% of the central incisors. Follicle size did not play a major role in influencing impacted canine position. The alveolus was narrower at the impacted canine side compared with the erupted canine side; however, the width of the alveolus on the impacted canine side is independent of the deciduous canines. A correlation was found between the proximity of the impacted canine to the incisors and their resorption. There was no common location where eruption was arrested, and great variation in the inclination of the impacted canine was found. CONCLUSIONS: 3D volumetric imaging of impacted canines can show the following: presence or absence of the canine, size of the follicle, inclination of the long axis of the tooth, relative buccal and palatal positions, amount of the bone covering the tooth, 3D proximity and resorption of roots of adjacent teeth, condition of adjacent teeth, local anatomic considerations, and overall stage of dental development. In short, 3D imaging is clearly advantageous in the management of impacted canines.

Adolescent↗

Randomized, double-blind study on effectiveness of intra-alveolar chlorhexidine gel in reducing the incidence of alveolar osteitis in mandibular third molar surgery.

Chlorhexidine (CHX) mouthwash is a good prophylactic agent for postextraction alveolar osteitis (AO), a very common complication. Recently, a bio-adhesive gel to deliver the active substance, CHX, has been introduced. The intra-alveolar positioning of the bio-adhesive gel allows more direct and prolonged action of CHX, which could be useful in the prevention of AO following extraction of impacted third molars. Presented here is a double-blind, randomised and parallel-group study evaluating the efficacy of the bio-adhesive 0.2% CHX gel (n=53) applied once within the alveolus during third molar surgery, compared to placebo gel only (n=50), in reducing the incidence of impacted third-molar postextraction AO. A 63.33% reduction in the occurrence of AO (P=0.019) was observed in the experimental group. In the control group, the occurrence of AO was 30% compared to 11% in the experimental group. Bio-adhesive 0.2% CHX gel, applied only once within the alveolus site at the time of surgery, may reduce the incidence of AO following removal of impacted third molars.

Adolescent↗

Interpretation of chemically created periapical lesions using direct digital imaging.

Perchloric acid (70%) was used to create simulated periapical lesions in tooth sockets of 15 dentate cadaver jaw specimens. Using the Trophy USA direct digital radiographic system, linear images were captured at selected time intervals after initial acid application and altered by contrast reversal, pseudocolor enhancement, and two forms of histogram equalization. The 525 total images were randomized for display on a computer monitor for evaluation by five endodontists. Images were evaluated twice by each rater, with viewings 1 to 2 wk apart. Statistical analysis determined interrater variability, intrarater reproducibility, and the relative merits of each enhancement technique. At 8, 12, 16, and 24 h after acid application, both techniques of histogram equalization yielded a statistically significant improvement over reverse contrast in perception of periapical patholais. Linear and pseudocolor-enhanced images were also significantly more diagnostic than reverse contrast at 12, 16, and 24 h. Intrarater reproducibility showed moderate agreement, but analysis showed only a fair level of interrater agreement.

Alveolar Bone Loss↗

A comparative investigation on autologous mandibular and iliac crest bone grafts. An experimental study in sheep.

Nine Friesian sheep (age 1.5-2 years) were used to evaluate the quality of bone repair in artificial cleft-like maxillary defects repaired with autologous bone grafts of embryologically different origin. After bilateral extraction of the upper first deciduous molar the tooth socket was enlarged resulting in an open connection between the oral and nasal cavities. In the same session a graft, obtained from the iliac crest, was implanted in one of the two bilateral maxillary defects and on the opposite side a graft, obtained from the mandible, was inserted. During the experiment, serial radiographs were taken. After six months the sheep were sacrificed and the bone of the grafted areas was submitted to histological and histomorphometric investigation. Comparing both areas, it appeared that the quality of bone remodelling and alveolar reconstruction was independent of the origin of the grafts.

Alveolar Process↗

Effects of projection geometry and number of projections on accuracy of depth discrimination with tuned-aperture computed tomography in dentistry.

OBJECTIVE: The purpose of this study was to determine the degree to which the number and angular disparity of component projections influence depth discrimination with tuned-aperture computed tomography. STUDY DESIGN: Groups of three tiny steel spheres served as fiducial references on and in four partially edentulous mandibles. Two spheres were attached to the facial and lingual surfaces of each mandible, and the third was fixed in the apical region of an open tooth socket. Errors in estimates of the depth of the apically positioned sphere relative to the other two spheres were determined from three-dimensional tuned-aperture computed tomography reconstructions. These data were compared with actual measurements produced independently with an optical micrometer. Multiple projections required by the tuned-aperture computed tomography reconstruction algorithm were produced from radially symmetric exposures bearing angular disparities of 5, 15, 30, and 45 degrees. The number of symmetrically dispersed projections per tuned-aperture computed tomography reconstruction likewise was varied systematically (2, 4, 8, 12, and 16 projections). These variables were manipulated through the use of a balanced factorial design. Depth estimates were performed by trained observers; the estimates were based on the determination of tuned-aperture computed tomography slices perceived as imaging the respective apical spheres in sharpest focus. Specimen and observer effects were also considered as independent variables. Resulting data were normalized by logarithmic transformation and analyzed statistically by analysis of variance. RESULTS: Significant differences (p < 0.005) were demonstrated for angular disparity and specimen effects, but the number of projections and the effect of the observer were not found to be statistically significant. CONCLUSIONS: In dentistry, angular disparities of 15 degrees or greater should be used when tuned-aperture computed tomography is being applied to diagnostic tasks requiring maximal depth discrimination accuracy.

Algorithms↗

Periodontal ligament cells secrete the factor that inhibits osteoclastic differentiation and function: the factor is osteoprotegerin/osteoclastogenesis inhibitory factor.

The periodontal ligament, a highly specialized connective tissue situated between the tooth and the alveolar bone of the tooth socket, has been thought to influence the remodeling of the alveolar bone. The effects of two human periodontal ligament fibroblastic cell populations (HPLFs) on osteoclast-like cell (OCL) formation and the function of authentic osteoclasts were examined. The addition of the conditioned media (CM) from both HPLF cultures (HPLF-CMs) to mouse bone marrow culture inhibited OCL formation in spite of the presence of 10(-8)M 1alpha, 25 dihydroxyvitamin D3 (1alpha,25(OH)2D3). This inhibitory effect was most remarkable when both CMs were added during day 6 to day 9 following bone marrow culture, just at the late stage of OCL differentiation. HPLF-CMs also induced a significant decrease in the pit area and the pit number formed by authentic osteoclasts on ivory slices. The administration of neutralizing monoclonal antibody (OI-1) against human osteoprotegerin (OPG)/osteoclastogenesis inhibitory factor (OCIF) with HPLF-CMs to mouse bone marrow culture almost completely blocked the inhibitory effect of these CMs on OCL formation. Immunofluorescent examination of HPLF with OI-1 revealed intense positive reactivity in the cytoplasm. Western blot analysis of HPLF-CM using anti-human OPG/OCIF polyclonal antibody resulted in the detection of bands of 60 kDa and 120 kDa which were consistent with those of OPG/OCIF. These results suggest that HPLF cells produce and secrete OPG/OCIF, and that this factor from HPLF prevents the differentiation of the late preosteoclast and the function of the mature osteoclasts.

Adolescent↗

The effect of streptozotocin-induced experimental diabetes mellitus on calvarial defect healing and bone turnover in the rat.

Tooth socket healing is delayed in diabetes mellitus due to impairment of the healing process. One reason for the poor healing may be an abnormal vascular response. The object of our experiments was to study the effect of diabetes mellitus on bone healing using a calvarial wound. Streptozotocin, injected intraperitoneally, was used to induce diabetes in rats. Both insulin-treated, streptozotocin-dosed animals and normal rats were used as controls. Bone formation was measured in the diabetic femur and tibia, and healing of bone defects by guided tissue regeneration was assessed. Cancellous bone volume and bone formation in the femur were greatly reduced in the diabetic model, indicating either a defect of mineralization or osteoid formation. The length, dry weight, ash weight and calcium content of the tibiae of diabetic rats were significantly less than those of the control groups. In a second experiment, a sterile wound was made in the calvaria of diabetic rats, and covered internally and externally with Gore-Tex membrane. Exuberant formation of a primitive bone was evident, with little evidence of osteoclastic resorption of the necrosed bone ends. This was despite the impaired bone formation observed in the long bones in the first experiment.

Animals↗

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.

Alveolar Process↗

Grafting mandibular third molar extraction sites: a comparison of bioactive glass to a nongrafted site.

OBJECTIVE: The purpose of this study was to evaluate the effectiveness of bioactive glass used as a graft material for regeneration of bone after the removal of impacted third molars. The healing distal to the second molars was followed, with documentation of the level of radiographic osseous fill and measurement of the changes in clinical attachment levels. METHODS: Fourteen (5 male and 9 female) patients completed the study at 1 year. After surgical removal of bilateral impacted third molars, one site was grafted with bioactive glass particles (BioGran, Orthovita, Malvern, Penn). The patients were followed up at 1 and 2 weeks after surgery for signs or symptoms of infection or any other complications that may have been related to the surgical procedures. At 3, 6, and 12 months, the clinical attachment levels distal to the second molars were assessed. At 6 and 12 months, radiographs were obtained to assess the amount of osseous fill distal to the second molars. RESULTS: A split-mouth design was used to test the efficacy of the grafting materials. Clinical attachment levels in the grafted sites were significantly higher than in the nongrafted sites. There was not a significant increase in bone formation when comparing the 2 sides at 1 year. CONCLUSION: This study suggests that treatment of third molar extraction sites with bioactive glass does significantly alter the clinical attachment level but not the level of osseous fill distal to the second molars after 1 year.

Adult↗

Botryomycosis: first report of human brain involvement.

A 57-year-old man, previously well except for dental caries and a history of Wolff-Parkinson-White syndrome, presented with marching right-sided motor seizures of sudden onset. Cerebral arteriography and scan demonstrated an avascular left frontal lobe mass. At operation, it was identified as an abscess and was totally excised. Histologically, granules resembling those seen in actinomycosis were demonstrated in the abscess wall, but special stains revealed gram-positive cocci in chains within and outside the granules. Fungal spores, mycelia, or branching filamentous structures were absent. Acid-fast stains were negative, and alpha-hemolytic streptococcus was recovered in pure culture from the abscess. While the patient was convalescing with penicillin therapy, a dental survey revealed the presence of periodontal abscesses which were drained by exodontia. Culture of the tooth sockets showed alpha-hemolytic streptococcus and Staphylococcus aureus. The literature on this relatively rare bacterial disease which histologically resembles actinomycosis is reviewed.

Actinomycosis↗

Expression of TGF-beta isoforms and their receptors during mineralized nodule formation by rat periodontal ligament cells in vitro.

Transforming growth factor-betas (TGF-beta s) and bone morphogenetic proteins (BMPs), members of a TGF-beta superfamily, are known to play an important role in osteogenic cell differentiation and consequently bone formation. We have reported previously that periodontal ligament (PDL) cells differentiate and form mineralized nodules when cultured in the presence of dexamethasone (Dex), beta-glycerophosphate (GP) and ascorbic acid (AA). To understand the roles of TGF-beta isoforms (TGF-beta 1, 2 and 3) and TGF-beta type I receptors (activin receptor-like kinase (ALK)-2, -3, -5 and -6) in PDL cell differentiation, their expression was investigated using Northern blot analysis. Rat PDL cells, derived from coagulum in the tooth socket, were cultured in the presence of Dex (5 microM), GP (10 mM) and AA (50 micrograms/ml) for up to 21 d. Total RNA was isolated from PDL cells after 0, 7, 14 and 21 d and used for northern blot analysis of mRNAs for matrix proteins, TGF-beta isoforms and their receptors using 32P-labeled cDNAs as probes. Four stages showing distinct morphological characteristics and matrix expression during development of mineralized nodules were identified. Type I collagen (Col I) and SPARC (secreted protein, acidic and rich in cysteine) mRNAs were expressed at the confluent stage, but decreased during the mineralization stage. Osteopontin (OPN) and alkaline phosphatase (ALP) transcripts were initially observed at multilayer stage, while bone sialoprotein (BSP) and osteocalcin (OC) at the nodule stage and all 4 were expressed thereafter. TGF-beta 1 mRNA expression increased with the progression of PDL cell differentiation, while a relatively high level of TGF-beta 3 transcript decreased slightly during their differentiation. TGF-beta 2 mRNA was not expressed. The expression of TGF beta-RI mRNA decreased, whereas that of TGF beta-RIII increased dramatically with PDL cell differentiation. TGF beta-RII gene activities remained high throughout all stages. ALK-2, ALK-3 and ALK-6 mRNA expression increased with the progression of PDL cell differentiation, suggesting that these receptors may play important roles in Dex-induced PDL cell differentiation and mineralized nodule formation.

Animals↗

Evaluation of fetal midface anatomy related to facial clefts: use of US.

PURPOSE: To identify the sonographic appearance of normal fetal midface anatomy of in vitro fetal specimens and to correlate the appearance with that of in utero fetuses to determine which aspects of this anatomy can be evaluated clinically. MATERIALS AND METHODS: The midface structures of 12 normal fetal specimens were examined in the axial, sagittal, and coronal planes with ultrasound. The results were correlated with those of other modalities. One hundred consecutive, normal in utero fetuses were scanned in the same planes in an attempt to identify the same anatomic landmarks identified in the fetal specimens. RESULTS: In the fetal specimens, the upper lip (coronal plane), alveolar ridge, tooth sockets, point of fusion of primary and secondary palates (axial plane), and fusion line of the secondary palate (sagittal plane) were consistently observed. In the 100 in utero fetuses, the same anatomy was visualized in the coronal plane in 95 (95%), in the axial plane in 97 (97%), and in the sagittal plane in 26 (26%). The mean scanning time was 2.8 minutes. CONCLUSION: Important aspects of fetal midface anatomy that relate to facial clefts can be observed in utero at prenatal sonography in the coronal and axial planes.

Cleft Lip↗

Lead exchange in teeth and bone--a pilot study using stable lead isotopes.

Stable lead isotopes and lead concentrations were measured in the enamel and dentine of permanent (n = 37) and deciduous teeth (n = 14) from 47 European immigrants to Australia to determine whether lead exchange occurs in teeth and how it relates to lead exchange in bone. Enamel exhibits no exchange of its European-origin lead with lead from the Australian environment. In contrast, dentine lead exchanges with Australian lead to the extent of approximately 1% per year. In one subject, trabecular bone from the tooth socket exchanged almost all its European lead with Australian lead over a a 15-year period (turnover of approximately 6% per year), similar to the approximately 8% per year proposed for lead turnover in trabecular bone. The repository characteristics of intact circumpulpal dentine were investigated by analyses of four sets of contiguous slices from six teeth: 1) a set consisting of slices with intact circumpulpal dentine and cementum; 2) a set in which these areas were removed; 3) another set consisting of slices with intact circumpulpal dentine and cementum; and 4) a set without cementum. These analyses show relatively small differences in isotopic composition between contiguous slices except that circumpulpal dentine appears to be the dominant control on lead concentration. There is a significant correlation (R2 = 0.19, p = 0.01, n = 34) of dentine lead concentration and rate of exchange with residence time from the country of origin and Australian lead, but there is no such correlation with enamel lead concentration. Analyses of permanent and deciduous teeth of subjects from other countries who have resided in Australia for varying lengths of time should resolve some of the questions arising from this pilot study.

Adult↗

Management strategies of the unsalvageable tooth.

UNLABELLED: A tooth is deemed unsalvageable when it can no longer be preserved for use of function or aesthetic purposes. If proper treatment planning is done, tooth extraction can be performed with adjunctive procedures, to provide a more predictable restorative outcome for the patient. This review paper summarizes the possible treatment strategies available for the management of a tooth with a poor prognosis, so as to manage the patient better before the restorative treatment phase commences. CLINICAL RELEVANCE: The management of an unsalvageable tooth using interdisciplinary approaches offers the clinician strategies for providing a more favourable restorative outcome.

Alveolar Bone Loss↗