Bacterial colonization associated with fixed orthodontic appliances. A scanning electron microscopy study.

This investigation was undertaken to assess bacterial plaque accumulation adjacent to orthodontic brackets. Experiments were carried out on 11 subjects who were scheduled for orthodontic treatment including extraction of two or four premolars. Metal brackets were bonded to the premolars to be extracted using macro-filled bonding composite. A conventional elastomeric ring was placed around one bracket and a steel ligature wire around the bracket on the contralateral tooth. The subjects were told to continue their normal oral hygiene regimen. Teeth were extracted at 1, 2, or 3 weeks after bracket bonding. Scanning electron microscopic (SEM) examination of brackets, excess composite, and buccal enamel revealed that mature plaque was present on excess composite at 2 and 3 weeks after bonding, whereas plaque on the gingival enamel surface was still at an early stage of development. The results demonstrate that excess composite around the bracket base is the critical site for plaque accumulation due to its rough surface and the presence of a distinct gap at the composite-enamel interface. The method of ligation does not appear to influence the bacterial morphotypes on both composite and enamel surfaces.

Periodontal repair in dogs: effect of rhBMP-2 concentration on regeneration of alveolar bone and periodontal attachment.

The objective of this study was to evaluate the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) concentration on regeneration of alveolar bone and cementum, and on associated root resorption and ankylosis. Contralateral, critical size, supra-alveolar, periodontal defects were surgically produced and immediately implanted with rhBMP-2 in an absorbable collagen sponge (ACS) carrier in 8, young adult, male, beagle dogs. 6 animals received rhBMP-2/ACS (rhBMP-2 at 0.05, 0.10, or 0.20 mg/mL; total construct volume/defect approximately 4.0 mL) in contralateral defects following an incomplete block design. 2 animals received rhBMP-2/ACS (rhBMP-2 at 0 and 0.10 mg/mL) in contralateral defects (controls). The animals were euthanised at 8 weeks post-surgery and block sections of the defects were collected for histologic and histometric analysis. Supra-alveolar periodontal defects receiving rhBMP-2 at 0.05, 0.10, or 0.20 mg/ml exhibited extensive alveolar regeneration comprising 86%, 96%, and 88% of the defect height, respectively. Cementum regeneration encompassed 8%, 6%, and 8% of the defect height, respectively. Root resorption was observed for all rhBMP-2 concentrations. Ankylosis was observed in almost all teeth receiving rhBMP-2. Control defects without rhBMP-2 exhibited limited, if any, evidence of alveolar bone and cementum regeneration, root resorption, or ankylosis. Within the selected rhBMP-2 concentration and observation interval, there appear to be no meaningful differences in regeneration of alveolar bone and cementum. There also appear to be no significant differences in the incidence and extent of root resorption and ankylosis, though there may be a positive correlation with rhBMP-2 concentration.

Effect of allogeneic freeze-dried demineralized bone matrix on regeneration of alveolar bone and periodontal attachment in dogs.

This split-mouth study was designed to evaluate regeneration of alveolar bone and periodontal attachment following implantation of allogeneic, freeze-dried, demineralized bone matrix (DBM). Buccal fenestration defects (6x4 mm) were created on the maxillary canine teeth in 6 beagle dogs. DBM was implanted into one randomly selected defect in each animal. The contralateral defect served as surgical control. Tissue blocks were harvested following a 4-week healing interval and prepared for histometric analysis. DBM was discernible in all implanted defects with limited evidence of bone metabolic activity. The DBM particles appeared invested within a dense connective tissue, often in close contact to the instrumented root. Fenestration defect height averaged 3.8+/-0.1 and 3.7+/-0.3 mm, total bone regeneration 0.9+/-0.9 and 0.4+/-1.2 mm, and total cementum regeneration 2.3+/-1.5 and 0.6+/-0.7 mm for DBM and control defects, respectively. Differences with regards to cementum regeneration were statistically significant (p=0.03). In summary, the results of this study suggest that DBM implants may enhance cementum regeneration in this defect model, and that they have no apparent effect on alveolar bone regeneration. Enhanced cementum regeneration may be possibly be explained by provisions for guided tissue regeneration from the implant suppressing a significant influence of the gingival connective tissue on the healing process. Moreover, a 4-week healing interval appears insufficient for turnover of DBM.

Effect of allogenic freeze-dried demineralized bone matrix on guided tissue regeneration in dogs.

This randomized, split-mouth study was designed to evaluate the adjunctive effect of allogenic, freeze-dried, demineralized bone matrix (DBM) to guided tissue regeneration (GTR). Contralateral fenestration defects (6 x 4 mm) were created 6 mm apical to the buccal alveolar crest on maxillary canine teeth in 6 beagle dogs. DBM was implanted into one randomly selected fenestration defect. Expanded polytetrafluoroethylene (ePTFE) membranes were used to provide bilateral GTR. Tissue blocks including defects with overlying membranes and soft tissues were harvested following a four-week healing interval and prepared for histometric analysis. Differences between GTR+DBM and GTR defects were evaluated using a paired t-test (N = 6). DBM was discernible in all GTR+DBM defects with limited, if any, evidence of bone metabolic activity. Rather, the DBM particles appeared solidified within a dense connective tissue matrix, often in close contact to the instrumented root. There were no statistically significant differences between the GTR+DBM versus the GTR condition for any histometric parameter examined. Fenestration defect height averaged 3.7+/-0.3 and 3.9+/-0.3 mm, total bone regeneration 0.8+/-0.6 and 1.5+/-0.8 mm, and total cementum regeneration 2.0+/-1.3 and 1.6+/-1.7 mm for GTR+DBM and GTR defects, respectively. The histologic and histometric observations, in concert, suggest that allogenic freeze-dried DBM has no adjunctive effect to GTR in periodontal fenestration defects over a four-week healing interval. The critical findings were 1) the DBM particles remained, embedded in dense connective tissue without evidence of bone metabolic activity; and 2) limited and similar amounts of bone and cementum regeneration were observed for both the GTR+DBM and GTR defects.

Microvascular system of the rat incisor enamel organ. A scanning electron microscopic study of vascular corrosion casts.

The rat incisor is a commonly used model in studies of tooth eruption, amelogenesis and effects of mechanical loading on the dental and periodontal tissues. The purpose of this study was to assess the three-dimensional architecture of the microvascular bed of the rat incisor enamel organ, to describe the direction of blood flow, and to provide a histometric assessment of the vascular categories that can be statistically analyzed. Vascular corrosion casts were prepared and examined by scanning electron microscopy. The microvasculature of the labial periodontal space was arranged in three distinct layers. The inner layer in direct relation to the enamel organ consisted of a capillary network which was drained by short venules at the cemento-enamel junction. The intermediate layer consisted of arterioles oriented parallel to the long axis of the incisor mainly mid-labially, branching off smaller arterioles to the capillary network. The outer layer was formed by flattened sinusoid vessels of larger caliber. Blood supply was from the anterior superior alveolar artery branches through the arterioles into the capillary network. Drainage was postero-laterally along the cemento-enamel junction via short venules which emptied into the sinusoid vessels, finally to flow through Volkmann's canals into the alveolar bone via small venules. The findings demonstrate that the microvasculature of the rat incisor enamel organ has an exceptionally high level of physiologically-adapted structural organization.

Oral tissue reactions to suture materials.

Tissue reactions to natural and synthetic braided and monofilament suture materials in gingiva and oral mucosa were studied. A total of 138 sutures made of four commonly used materials were placed in the edentulous ridges and vestibular mucosa of eight beagle dogs. Biopsy specimens including the suture loop and surrounding tissues were obtained after 3, 7, and 14 days and processed for histologic analysis. The inflammatory reaction was more rapid and intense than the reaction that has been reported after suture placement in skin. Bacterial invasion of the suture track was a common sequela regardless of the material used, but it was particularly prominent for silk. The formation of a perisutural epithelial sleeve was well under way at 3 days and in some instances included the entire suture track within 7 days. Connective tissue reactions consisted of several well-defined, concentric perisutural zones. At 14 days, these zones were partly replaced by granulation tissue surrounded by a fibrous capsule. The synthetic monofilament suture elicited a mild inflammatory tissue response. The results showed that sutures placed in gingiva and oral mucosa produce a prolonged tissue response that is most likely a result of the continual influx of microbial contamination along the suture channel, which may be a lesser problem when sutures are placed in other surgical compartments. The results indicate that chromic gut sutures are rapidly and unpredictably absorbed when used in an environment characterized by moisture and infectious potential.

A correlation study of inflammatory cell mobilization in response to subgingival microbial colonization.

This study evaluated site-by-site the relations between subgingival microbial colonization and gingival tissue reactions. Experimental, deep periodontal defects were established at buccal surfaces of mandibular and maxillary canine teeth in 5 beagle dogs. The root surfaces were instrumented by a flame-shaped, fine-grained, rotating diamond point, or by a sharp curet. Following a 10-day postsurgical healing period, the dogs were fed a plaque-inducing diet for 70 days. The animals were then sacrificed and tissue blocks of the experimental sites including teeth and periodontal tissues were secured. The buccal gingiva was removed and processed for histomorphometric analysis while the teeth were prepared for scanning electron microscopic evaluation of the extent of subgingival microbial colonization. The results revealed that inflammatory cell density in the junctional epithelium and in the connective tissue were positively correlated to subgingival microbial colonization (P < 0.01). Furthermore, the degree of significance decreased with increasing distance from the plaque. The present study demonstrates that a close relation may exist between the extent of subgingival microbial colonization and inflammatory gingival tissue reactions.

Variability in fluoride content of subsurface water reservoirs.

Groundwater may contain high concentrations of fluoride. In most countries, however, information on the fluoride content is scarce and anecdotal. The aim of the present study was to make a comprehensive assessment of F- in the groundwater of a representative area of Norway, thereby establishing a more solid basis for appropriate health counseling. Relevant technical information was collected, together with water samples from 1063 underground water sources in 31 municipalities in the county of Hordaland. One thousand and two water samples were analyzed for F- and pH with an F(-)-selective electrode and a pH electrode, respectively. Mean F- was 0.30 mg/l (range, < 0.02-9.48). Fourteen per cent of the wells contained water with F- levels > or = 0.50 mg/l. In three municipalities well water had a mean F- concentration > 0.07 mg/l; in one instance the mean was as high as 1.45 mg/l. In 10 municipalities maximum F- values were > 1.50 mg/l. F- values showed a positive correlation with the pH of the water and the depth of the wells (P < 0.01) and a negative correlation with the age of the well (P < 0.05). The results indicated that low-capacity wells deliver water with a higher F- value than high-capacity wells. This trend, however, was not statistically significant. The results show that high-F groundwater is prevalent and emphasize that information on domestic water supply must be available before supplementary fluoride is prescribed for caries prophylactic purpose.

Wound healing after mucoperiosteal surgery in the cat.

The purpose of this study was to examine possible tissue-dependent differences in rate of healing after mucogingival flap surgery. After intrasulcular incision and a vertical-releasing incision distal to the maxillary and mandibular cuspids, buccal, full-thickness mucogingival flaps were raised in four quadrants of 10 adult cats. The triangular flaps were left open for 30 min and then repositioned and sutured. Tissue reactions were studied histologically after 1, 3, 7, 14, and 28 days of healing. Although new collagen occasionally was observed in the wound space in the free gingiva at 3 days, collagenous union between the cut dentogingival fibers and the flap seemed well established at 7 days. Flap reattachment to the denuded cortical bone was seen at 14 days in the region of the attached gingiva. In the region of the alveolar mucosa, however, residual coagulum and inflammatory reaction was present as late as at 28 days in several specimens. These observations indicate a marked difference in rate of healing among the different interfaces involved. These variations seem to be related to variations in size of the resulting wound space when a full-thickness mucoperiosteal flap is readapted over cervical root surfaces, alveolar bone crest, and denuded cortical bone, respectively.

Cemental tear: a risk factor in periodontal attachment loss.

The objective of this retrospective study was to evaluate the influence of cemental tear as a risk factor in periodontal attachment loss. Seventeen extracted, single-rooted teeth showing loss of attachment and having one cemental tear surface and one opposite intact surface were examined. The teeth were stained in 0.1% toluidine blue to visualize attached periodontal ligament remnants and examined in a light microscope under incident light. On each tooth, loss of attachment was measured along the long axis of the root from the cemento-enamel junction to the most coronal level of the periodontal ligament on intact as well as on defect surfaces. Cemental tear surfaces demonstrated a significantly greater loss of attachment than opposite intact surfaces (P < 0.0001). In one specimen, the cemental tear fragment was partially attached to the root after the extraction procedure. This specimen was processed for light microscopy to determine the location of the cemental tear fracture. Histological examination clearly revealed that the split between the root and the fragment had occurred along the cemento-dentinal border. The results indicate that cemental tear should be considered as a possible etiologic entity in localized rapid periodontal breakdown.

Atypically located paramolar. Report of a rare case.

A paramolar mesial to the mandibular second molar would be predicted from the atavistic theory on phylogenetic evolution of the human dentition. The actual finding of this feature, however, appears to have been reported only once. A 4 mm long tooth was found mesial to the right mandibular second molar in a 54-yr-old Japanese female. The tooth had a ball-shaped crown, a relatively normal crown/root length ratio, and a tapering root located entirely in the gingival soft tissue.

Influence of tooth instrumentation roughness on gingival tissue reactions.

Histological studies have demonstrated a relationship between the amount of subgingival plaque and the magnitude and extension of gingival tissue reactions. The objective of the present study was to evaluate inflammatory reactions in the gingival tissues facing plaque accumulation at a diamond and curet-instrumented root surfaces. Experimental, deep periodontal defects were established at buccal surfaces of mandibular and maxillary canine teeth in 5 beagle dogs. The root surfaces were instrumented by a flame-shaped, fine-grained. rotating diamond point, or by a sharp curet. Next, the dogs were fed a plaque-inducing diet for 70 days. The animals were then sacrificed, and tissue blocks of the experimental sites including teeth, alveolar bone, and gingival tissues were secured. The gingival soft tissue was processed for histomorphometric analyses at 3 levels. Epithelium and connective tissue area measurements showed no differences between the two instrumentations. Junctional epithelium (JE) cell point counts exhibited a higher proportion of inflammatory cells (IC)in specimens facing diamond compared to curet-instrumented defects. A higher proportion of IC was present within the coronal compared to the apical aspect of the JE for both instrumentations (P < 0.05). A significant difference in IC density between instrumentations was detected for non-infiltrated (P < 0.05), as well as for infiltrated (P < 0.01) connective tissue. The infiltrated connective tissue (ICT) inflammatory cell density was significantly (P < 0.01) and positively correlated to the JE inflammatory cell density (r = 0.75), and to area measurements of ICT (r = 0.55). The overall results demonstrate that the character of subgingival root instrumentations significantly affects gingival inflammatory reactions, most likely by influencing subgingival plaque formation.

Does root surface conditioning with citric acid delay healing?

Effects of topical citric acid application on tissue maturation was studied in standardized periodontal defects in 6 beagle dogs. Following elevation of facial mucoperiosteal flaps, fenestration defects, 3 mm in diameter, were made through the cortical bone and recessed 0.5 mm into the dentin of maxillary canines. 1 defect in each dog was conditioned with a saturated solution of citric acid for 3 min and then rinsed with saline. Control defects in contralateral teeth were treated with saline only for the same length of time. The defects were covered with an expanded polytetrafluoroethylene membrane and the flaps repositioned and sutured. 14 days postsurgery, healing appeared more advanced along the defect walls and floor than in the center of the defect in all instances. Histometrically, citric acid-conditioned defects exhibited a higher density of collagen fibers along the defect walls and floor and adjacent to the barrier membrane as well as more advanced resolution of the residual blood clot than the surgical controls. Differences in fibroblast density within specimen pairs were non-significant. All control defects but none of the acid-conditioned defects showed an artifactual split between the dentin walls and the granulation tissue. This study failed to support the contention that topical application of citric acid to root surfaces may delay healing following periodontal surgery.

Intra - and inter-examiner reproducibility in constant force probing.

This study evaluated intra- and inter-examiner reproducibility for a conventional manual probe versus a computer-interfaced force-controlled periodontal probe. 2 examiners recorded probing depths (PD) and relative attachment levels (AL) at 1128 sites in 15 periodontal maintenance patients. Each site was evaluated 2x, 7 to 10 days apart by both examiners. Probing force for the electronic probe was 15 g. PD intra-examiner reproducibility (within +/- 1.0 mm) for shallow sites (PD < or = 3 mm) was 98.6% versus 91.5% for the conventional versus the electronic probe for examiner 1 and 98.5% versus 88.7% for examiner 2. Corresponding values for deeper sites (PD > 3 mm) were 96.4% versus 85.9% for examiner 1 and 95.1% versus 77.0% for examiner 2. Generally, AL intra-examiner reproducibility was 1 to 3% lower than for PD. PD inter-examiner reproducibility (within +/- 1.0 mm) was 99.2% versus 90.7% for the conventional versus the electronic probe, respectively, for shallow sites and 95.4% versus 76.9% for deeper sites. AL inter-examiner reproducibility (within +/- 1.0 mm) was 1 to 5% lower than for PD. Both intra- and inter-examiner reproducibility was higher for anterior than for posterior sites. Mean PD and AL were similar for both examiners. However, the electronic probe consistently recorded 0.1 to 0.2 mm higher values than the conventional probe. Standard deviations indicated a greater variability for electronic than for manual probing. The results suggest that intra- and inter-examiner reproducibility may not necessarily be higher with an electronic, force-controlled periodontal probe than with a conventional manual probe.

Microvascular adaptation to transmucosal implants. A scanning electron microscopic study in the rat.

The profuse gingival microvasculature surrounding natural teeth has a unique architecture that contributes to the biologic seal and the anti-infective defense in the sulcular region. This study tested the hypothesis that this characteristic vascular arrangement is not replicated when gingival tissues adapt to a transmucosal implant. In 20 4-week-old Wistar rats, the upper first molar was extracted bilaterally and the gingiva at this site excised. Four weeks later, a 5-mm-long titanium screw was implanted at the extraction site on one side. After another 4 weeks, the rats were injected with liquid plastic resin through the carotid arteries and vascular casts prepared for scanning electron microscopy. The results showed that within 4 weeks after implant insertion, the microvasculature of the regenerated soft tissues immediately surrounding the implant had formed a characteristic arrangement that consisted of a dense subepithelial network as well as an orderly array of vertically oriented capillary loops. The density and diameter of the capillaries varied within a wide range, possibly related to the extent of healing and/or inflammatory conditions. On the contralateral, edentulous side, the capillaries formed a flattened meshwork showing a conspicuous absence of capillary loops. This study indicates that, when a transmucosal implant is inserted in an edentulous area, the mucosal microvasculature adapts and forms an arrangement very similar to that surrounding natural teeth.

Dynamics of wound healing in periodontal regenerative therapy.

Experimental animal and clinical studies have increased understanding of the biology of healing following periodontal regenerative therapy and factors that may influence the outcome. Formation of a new connective tissue attachment rather than a long junctional epithelium at the tooth-gingival flap interface is dependent upon the completion of a series of interactions among the root surface, plasma and tissue factors, and the connective tissue of the gingival flap. First, plasma proteins must adsorb to and remain in undisturbed contact with a generally noncompromised root surface. Next, adhesion of the established fibrin clot to the root surface must remain intact. Within days, a cellular and fibrous attachment will form; however, the tooth-gingival flap interface will still be vulnerable to wound-rupturing forces. Within two weeks the interface may have gained sufficient mechanical strength to offset such forces, at least in limited periodontal defects. Eventually, maturation of the fibrous attachment, including bone and cementum formation, will occur, particularly following adequate space provision by barrier membranes (guided tissue regeneration). A novel research focus involves how advances in molecular biology can translate to periodontal regenerative therapy. Bone morphogenetic proteins (BMPs) have been shown to support healing in a variety of skeletal sites. Recent research has demonstrated clinically significant alveolar bone and cementum regeneration with the use of a recombinant human BMP-2 implant. Such data suggest that growth-promoting substances, such as rhBMP-2, may significantly enhance periodontal regeneration and that use of such substances may radically recast current periodontal regenerative therapy.

"Collagen adhesion" revisited.

Histologic studies of periodontal reconstructive therapies have repeatedly shown a zone of nondescript connective tissue adaptation, or collagen adhesion, to the root surface between the apical extension of the junctional epithelium and the coronal extension of identifiable regenerated cementum. To clarify the nature of this zone, supra-alveolar periodontal defects were created bilaterally in the mandibular premolar region in three beagle dogs, and exposed roots were instrumented to remove the cementum. Flaps were then coronally advanced and sutured. Histologic analysis after 12 weeks of healing showed instrumented root dentin not associated with junctional epithelium, new cementum, root resorption, or ankylosis in 27 of 60 root surfaces. In 10 of these specimens subsequently examined by transmission electron microscopy, collagen fibrils were generally oriented parallel to and in close proximity to the root. A mechanism of collagen attachment was, in fact, functioning at sites of collagen adhesion; these areas may validly be included in the histologic attachment level reported in studies of periodontal reconstructive therapy.

Effect of specimen preparation method on relative volume of the microvasculature--a light microscopic study in rat tongue.

The study of corrosion casts by scanning electron microscope is an established means of investigating three-dimensional microvascular architecture. However, concern has been raised that the volume of the vasculature is affected by the perfusion procedure. Ten rats (group A) were standard perfusion fixed, while 10 rats (group B) were perfused with acrylic resin after fixation. Vascular volume was assessed by computer-aided analysis of thin sections of tongue lamina propria. The results showed that vascular volume in lamina propria of rat tongue is 13.6 +/- 4.1% (mean +/- SD). No significant differences in vascular volume were found between group A and group B specimens. It can be concluded that plastic-perfused specimens may be as reliable as histologic sections in obtaining a true image of vascular systems. Thus, corrosion casts are suitable for histometric analysis of the microvasculature.