Scanning electron microscopy study of new bone formation following small and large defects preserved with xenografts supplemented with pamidronate-A pilot study in Fox-Hound dogs at 4 and 8 weeks.

The aim of the present study was to evaluate the feasibility of SEM and EDX microanalysis on evaluating the effect of porcine xenografts (MP3®) supplemented with pamidronate during socket healing. Mandibular second premolars (P2) and first molars (M1) were extracted from six Beagle dogs. P2 were categorized as small defects (SD) and M1 as large defects (LD). Four random groups were created: SC (small control defects with MP3®), ST (small test defects MP3®+pamidronate), LC (large control defects with MP3®), and LT (large test defects MP3®+pamidronate). At four and eight weeks of healing the samples were evaluated fisically through scanning electron microscopy (SEM), and chemical element mapping was carried out by Energy dispersive X-ray spectroscopy (EDX). After four weeks of healing, SEM and EDX analysis revealed more mineralized bone in ST and LT groups compared with control groups (p<0.05). After eight weeks, Ca/P ratios were slightly higher for small defects (groups SC and ST); in SEM description, in both control and test groups, trabecular bone density was similar to the adjacent mineralized cortical bone. Within the limitations of this experimental study, SEM description and EDX elemental microanalysis have demonstrated to be useful techniques to assess bone remodelling of small and large defects. Both techniques show increased bone formation in test groups (MP3® modified with pamidronate) after four and eight weeks of healing.

Test in canine extraction site preservations by using mineralized collagen plug with or without membrane.

The aim of this study was to discuss the feasibility of porous mineralized collagen plug and bilayer mineralized collagen-guided bone regeneration membrane in site preservation in extraction sockets. The third mandibular premolars on both sides were extracted from four dogs, thus there were 16 alveolar sockets in all dogs and were randomly assigned into three groups. Group A had six alveolar sockets, and groups B and C had five alveolar sockets, respectively. Each alveolar socket of group A was immediately implanted with a porous mineralized collagen plug and covered with a bilayer mineralized collagen-guided bone regeneration membrane after tooth extraction. Alveolar sockets of group B were implanted with porous mineralized collagen plug only, and group C was set as blank control without any implantation. The healing effects of the extraction sockets were evaluated by gross observation, morphological measurements, and X-ray micro-computed tomography after twelve weeks. Twelve weeks after operation, both groups A and B had more amount of new bone formation compared with group C; in terms of the degree of alveolar bone height, group A was lower than groups B and C with significant differences; the bone mineral density in the region of interest and bone remodeling degree in group A were higher than those of groups B and C. As a result, porous mineralized collagen plug could induce the regeneration of new bone in extraction socket, and combined use of porous mineralized collagen plug and bilayer mineralized collagen guided bone regeneration membrane could further reduce the absorption of alveolar ridge and preserve the socket site.

Evaluación de la osteogénesis pos extracción en el alvéolo de rata: un modelo experimental / Osteogenic process evaluation following rat molar tooth extraction: an experimental model

coObjetivo: analizar la eficacia de un modelo experimental desarrollado en el laboratorio para evaluar radiográfica e histológicamente el proceso de osteogénesis y reparación alveolar. Materiales y métodos: se utilizaron 10 (n=10) ratas Wistar macho de aproximadamente 90 gramos de peso. Luego de anestesiados, se les realizó la extracción de dos molares superiores. Se formaron 2 grupos experimentales de 5 (n=5) ratas cada uno. Los maxilares fueron removidos, separados en mitade y fijados en una solución de formol-buffer al 10 por ciento. Resultados: en el grupo I se observaron imágenes radiolúcidas de los alvéolos vacíos. En los cortes histológicos se observaron imágenes compatibles con hemorragia y fibras periodontales remanentes. En el grupo II se encontraron imágenes radioopacas intraalveolares compatibles con tejido óseo. Los cortes histológicos revelaron el desarrollo de trabéculas óseas y espacios medulares que ocupaban casi la totalidad del espacio alveolar. Conclusiones: el modelo propuesto permitiría evaluar los cambios morfológicos producidos durante la osteogénesis posextracción. Adicionalmente, facilitaría la evaluación de los efectos producidos por diferentes condiciones experimentales.

Chronological changes in the microstructure of bone during peri-implant healing: a microcomputed tomographic evaluation.

Our objectives were to examine the titanium-bone interfaces chronologically and to clarify the process of osseointegration using microcomputed tomography (microCT). The mandibular premolars of 3 dogs were extracted and 12 weeks later 2 Straumann dental implants were installed in each quadrant. The microstructural changes at the bone-implant interface at the first, fourth, and eighth weeks after installation were evaluated by microCT and a 3-dimensional image was constructed. The microstructural measurements at the bone-implant interface, including the trabecular number (Tb.N), bone volume fraction (BV/TV), structure model index (SMI), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) and connectivity density (Conn.D), were measured. Experimental outcomes were analysed for correlations between time and microstructural variables of trabecular bone. Tb.N increased significantly during the eighth week compared with that during the first and fourth week (p=0.001, 0.002). BV/TV increased generally with time and there were significant differences (p=0.003) between each time group. While Tb.Sp decreased with time and changed significantly during the eighth week compared with that during the first week (p=0.021), differences in SMI, Tb.Th and Conn.D did not differ significantly according to time of implant. The significant increase in the Tb.N and BV/TV during the eighth week after insertion of implants indicates that the formation of new bone before the eighth week is the key to osseointegration.

Particulate bone matrix usage for alveolar bone conservation. A histomorphometric study.

UNLABELLED: Different filling materials have been used in an attempt to repair bone loss situations. OBJECTIVE: The present study aimed to examine the effect of a bone matrix in post - extraction remodelling of the alveolar bone, and to perform a histomorphometric analysis of the residual alveolar ridges in Wistar rats. MATERIAL AND METHODS: Both rat first lower molars were extracted and the right alveoli were filled with particles of a bone matrix with mineral components (MO - UNC) (experimental group, EG). The left alveoli were used as a control group (CG). The animals were sacrificed at 0 hr., 15, 30 and 60 days after extraction, and the samples were processed. Histological sections were made at the level of the mesial alveolus of the first lower molar. Repair of the alveoli was histologically evaluated and a histomorphometric study of total alveolar volume (TAV), height of the buccal plate (Bh), height of the lingual plate (Lh) and percentage of osseointegration (OI) of the particles was performed to compare the residual ridges of CG with those of the EG. Statistical analysis of the data was performed. RESULTS: In the cases of the experimental group, newly - formed bone tissue was identified around the MO - UNC particles (osseointegration). Histomorphometric data indicate that, at 60 days post - extraction, TAV was significantly greater for EG when compared with CG (p <0.05) and the percentage of osseointegration of the particles increased as a function of time (57.6 %, 90.5% y 95.5%, for EG at 15, 30 y 60 days respectively). CONCLUSIONS: The bone matrix (MO - UNC) evaluated in this study is an osteoconductive material that prevents the collapse of post - extraction alveolar bone.

Particulate bone matrix usage for alveolar bone conservation. A histomorphometric study / Particulate bone matrix usage for alveolar bone conservation. A histomorphometric study.

UNLABELLED: Different filling materials have been used in an attempt to repair bone loss situations. OBJECTIVE: The present study aimed to examine the effect of a bone matrix in post - extraction remodelling of the alveolar bone, and to perform a histomorphometric analysis of the residual alveolar ridges in Wistar rats. MATERIAL AND METHODS: Both rat first lower molars were extracted and the right alveoli were filled with particles of a bone matrix with mineral components (MO - UNC) (experimental group, EG). The left alveoli were used as a control group (CG). The animals were sacrificed at 0 hr., 15, 30 and 60 days after extraction, and the samples were processed. Histological sections were made at the level of the mesial alveolus of the first lower molar. Repair of the alveoli was histologically evaluated and a histomorphometric study of total alveolar volume (TAV), height of the buccal plate (Bh), height of the lingual plate (Lh) and percentage of osseointegration (OI) of the particles was performed to compare the residual ridges of CG with those of the EG. Statistical analysis of the data was performed. RESULTS: In the cases of the experimental group, newly - formed bone tissue was identified around the MO - UNC particles (osseointegration). Histomorphometric data indicate that, at 60 days post - extraction, TAV was significantly greater for EG when compared with CG (p <0.05) and the percentage of osseointegration of the particles increased as a function of time (57.6

, for EG at 15, 30 y 60 days respectively). CONCLUSIONS: The bone matrix (MO - UNC) evaluated in this study is an osteoconductive material that prevents the collapse of post - extraction alveolar bone.

Particulate bone matrix usage for alveolar bone conservation. A histomorphometric study. / Particulate bone matrix usage for alveolar bone conservation. A histomorphometric study.

UNLABELLED: Different filling materials have been used in an attempt to repair bone loss situations. OBJECTIVE: The present study aimed to examine the effect of a bone matrix in post - extraction remodelling of the alveolar bone, and to perform a histomorphometric analysis of the residual alveolar ridges in Wistar rats. MATERIAL AND METHODS: Both rat first lower molars were extracted and the right alveoli were filled with particles of a bone matrix with mineral components (MO - UNC) (experimental group, EG). The left alveoli were used as a control group (CG). The animals were sacrificed at 0 hr., 15, 30 and 60 days after extraction, and the samples were processed. Histological sections were made at the level of the mesial alveolus of the first lower molar. Repair of the alveoli was histologically evaluated and a histomorphometric study of total alveolar volume (TAV), height of the buccal plate (Bh), height of the lingual plate (Lh) and percentage of osseointegration (OI) of the particles was performed to compare the residual ridges of CG with those of the EG. Statistical analysis of the data was performed. RESULTS: In the cases of the experimental group, newly - formed bone tissue was identified around the MO - UNC particles (osseointegration). Histomorphometric data indicate that, at 60 days post - extraction, TAV was significantly greater for EG when compared with CG (p <0.05) and the percentage of osseointegration of the particles increased as a function of time (57.6

, 90.5

y 95.5

, for EG at 15, 30 y 60 days respectively). CONCLUSIONS: The bone matrix (MO - UNC) evaluated in this study is an osteoconductive material that prevents the collapse of post - extraction alveolar bone.

Histology of tooth attachment tissues and plicidentine in Varanus (Reptilia: Squamata), and a discussion of the evolution of amniote tooth attachment.

Few recent studies have examined the histological basis for tooth attachment in squamates. In the past few years, a surge of interest in this topic has led to the intriguing suggestion that the major tissues derived from the tooth germ (enamel, dentine, cementum and alveolar bone), are conservative and are present in all amniotes. In this study, we describe the histology and development of the tooth attachment complex in Varanus rudicollis, the rough-neck monitor. We provide the first published evidence for the role of cementum and alveolar bone in tooth attachment in varanoid lizards. In Varanus, cementum is deposited on the external surface of the tooth root as well as at the base of the tooth, where it plays a role in the attachment of the tooth to the jawbone. Alveolar bone is also involved in tooth ankylosis. Our results support the hypothesis that the major tooth germ tissues are found in all amniotes. We provide insights into the structure and development of plicidentine, defined as infolding of the dentine around the tooth base. This feature is unique to varanoids among extant tetrapods and is the third tissue implicated in tooth attachment in Varanus. Plicidentine develops asymmetrically along the labial-lingual axis of a tooth. Varanus is characterized by the presence of both primary and higher-order lamellae, which anastomose to form a honeycomb-like surface that then interacts with the more basal attachment tissues.

In vivo graft performance of an improved bone substitute composed of poor crystalline hydroxyapatite based biphasic calcium phosphate.

The ability to promote new bone formation of poor crystalline hydroxyapatite (PC-HA) based biphasic calcium phosphate (BCP) bone substitutes were investigated. Various ratios of porous PC-HA/ß-TCP (70/30, 60/40, and 0/100) grafts were fabricated. SEM and XRD measurements were performed to study the morphology and crystalline structure. Cylindrical artificial bone defects (3 × 6 mm(2)) were produced in alveolar bone at premolars extraction site and then filled with sterilized bone grafts. Commercial MBCP and unfilled empty defect served as control groups. At 8 weeks postoperation, samples were harvested from each artificial defect site for histological analysis. New bone formation of all the PC-HA/ß-TCP groups was significantly greater than that of the empty control group (p<0.05), but without statistical difference from that of MBCP group. The degree of uniformity of new bone formation within defect region for PC-HA/ß-TCP (60/40) was higher than that for MBCP. The PC-HA/ß-TCP grafts showed enhanced bone regenerations with a more even dispersion of new bone formation than the other materials without causing inflammation, suggesting that these materials may be an alternative choice for bone void fillers in dental applications.

Virtual histology by means of high-resolution X-ray CT.

Micro-CT is a non-destructive technique for 3D tomographic investigation of an object. A 3D representation of the internal structure is calculated based on a series of X-ray radiographs taken from different angles. The spatial resolution of current laboratory-used micro-CT systems has come down over the last years from a few tens of microns to a few microns. This opens the possibility to perform histological investigations in 3D on a virtual representation of a sample, referred to as virtual 3D histology. The advantage of micro-CT based virtual histology is the immediate and automated 3D visualization of the sample without prior slicing, sample preparation like decalcification, photographing and aligning. This not only permits a drastic reduction in preparation time but also offers the possibility to easily investigate objects that are difficult to slice. This article presents results that were obtained on punch biopsies of horse skin, (dental) alveolus of ponies and chondro-osseous samples from the tarsus of foals studied with the new high resolution micro-CT set-up (HRXCT) at the Ghent University (Belgium) (http://www.ugct.ugent.be). This state-of-the-art set-up provides a 1 micron resolution and is therefore ideally suited for a direct comparison with standard light microscopy-based histology.

[Enforcing osseointegration of dental implantates spray-coated by bioceramics with the help of hyaluronic acid and hydroxyapatite gel in experimental conditions].

Influence of HAP-gel (2 g of 2% solution of hyaluronic acid mixed with 0,5 g of hydroxyapatite and 0,1 ml of colloidal silver) upon osseointegration in case of delayed introduction of titanium implantates in dog jaw. By scanning electron microscopy it was shown that solely use either of HAP-gel or of ceramic spraying increased direct contact area between bone and implantates in the 6 and 9 months time period. Combination of spray-coated ceramic with HAP-gel was effective in 3 months after implantation, when solely the HAP-gel or the ceramic spraying were little effective. In the following terms of experiment (6 and 9 months) significant differences between groups 3 and 4 (implantate with ceramic spraying but without HAP-gel in the alveolus and implantate with ceramic spraying and with HAP-gel in the alveolus) were not found. The area of implantate integration with jaw bone (cortical part of it was excluded) was equal to 80% and was maximal for the given conditions of the experiment.

Análisis del comportamiento de un material de relleno, colágeno bovino desmineralizado, en alvéolos post extracción: caso clínico / Analysis of the behavior of a filling material, demineralized bovine collagen, in postextraction sockets: a clinical case

La utilización de sustitutos óseos de diferente composición es una alternativa terapéutica de creciente aplicación en el área odontológica y traumatológica. En estudios previos (Histomorphometric evaluation of the effect of bovine collagen granules on bone healing. An experimental study in rats Agorustovich, FJVeinsten, ORCosta, MBGuglielmotti, Rev. AOL Vol. 17 Nº 1-2/2004/9-13 -Fig 1, Sustituto óseo de colágeno en la reparación ósea FJVeinsten, MSivak, ORCosta, MBGuglielmotti- SAIO 2006- Fig 2) describimos la respuesta del tejido óseo en un modelo experimental de reparación ósea en presencia de un biomaterial de colágeno, siguiendo una técnica previamente descripta (Guglielmotti et. al. J. Oral Maxillofac. Surg. 43:359, 1985). El objetivo del presente estudio fue evaluar la reparación ósea post-exodoncia en un caso clínico, ante la colocación de un relleno de matriz desmineralizada de hueso bovino, que permita la formación de un nuevo hueso para la posterior colocación de implantes oseointegrados.Se realizó la colocación del material en un alvéolo post extracción del sector de incisivos inferiores que presentaban extrema movilidad, por infección de origen periodontal. En dicha zona se realizó a los 90 días una biopsia por punción aspirativa, utilizando una aguja de Jam Schidi. El material obtenido fue fijado en formol al 20%, analizado microscópicamente luego de su descalcificación, inclusión en parafina y coloración con hematoxilina - eosina.Resultados: el análisis histológico de las muestras obtenidas, mostró la formación de nuevo hueso con presencia de osteoblastos en relación directa con un área de formación ósea, no encontrando focos de respuesta inflamatoria adversa que compliquen el cuadro de reparación. No se observaron reacciones adversas ni complicaciones en el post operatorio inmediato o mediato

Low frequency noise effects on the periodontium of the Wistar rat - a light microscopy study

Long-term low-frequency noise (LFN)(m500Hz, infrasound included) exposure is known to cause extracellular matrix proliferation with fibrosis, in the absence of inflammatory signs. The aim of this work was to study the morphological alterations to the periodontium of Wistar rats exposed to LFN. 10 rats were exposed to LFN for 2184 consecutive hours and another 10 were kept in silence. The mandibles were removed, kept in 10%buffered formalin, sectioned sagitally, stained with haematoxylin-eosin (HE) and Masson¡¯s trichromic solution (TCM) and observed with light microscopy (LM).The results revealed a disappearance of thecementum, irregular erosion of surface alveolarbone, and signs of bone necrosis, with detached bone particles. The periodontal ligament was disorganized and had deficient anchorage of the fibers.These findings may be due to a direct effectof noise and vibration impinging on the structures,to stress, to vascular alterations or to a combination of these factors. They may also explain the alterations in alveolar bone, reported by other authors (AU)

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Histomorphometric study on the effects of age on orthodontic tooth movement and alveolar bone turnover in rats.

The aim of this study was to investigate, in a rat model, the effects of age on the amount of tooth movement and concomitant changes in alveolar bone turnover activity adjacent to orthodontically treated tooth roots. Rats (n = 48) of four different age groups (10, 30, 50, and 80 wk of age) were used in the experiment. Maxillary first molars were tipped mesially with a nickel titanium alloy coil-spring for 2 wk by a continuous force of 10 cN. Forty-eight age-matched untreated rats were used as controls. The changes in alveolar bone turnover were assessed histomorphometrically. Two weeks after the start of tooth movement, the amount of tooth movement was found to decrease with age and was significantly different among the four age groups. The histomorphometric study demonstrated that, in all experimental groups, turnover of alveolar bone increased significantly compared with that of each age-matched untreated group. However, the rate of increase decreased in an age-related manner. These results suggest that the age-dependent decrease in alveolar bone turnover activity, in response to mechanical forces, may negatively affect the amount of tooth movement.

Relationship between porotic changes in alveolar bone and spinal osteoporosis.

Epidemiological studies have shown that post-menopausal women who do not use an estrogen supplement have fewer teeth than those who do. We hypothesized that changes in the dentition of post-menopausal women might be due to alveolar bone alterations by estrogen deficiency. To clarify this, we analyzed the microstructural alveolar bone changes in ovariectomized monkeys and compared these with their lumbar bone mineral density. The % of baseline bone mineral density showed a significant decrease in the ovariectomized group as compared with the controls. The second-molar interradicular septa in ovariectomized monkeys showed a significantly decreased nodes number, cortices number, and an increased structural model index value. More pores were seen in the ovariectomized group at the top of the septa. This study demonstrated that, in such monkeys, estrogen deficiency led to fragility of the trabecular structure of the molar alveolar bone, and such fragility was inversely correlated with lumbar bone mineral density.

The canalicular structure of compact bone in the rat at different ages.

Osteocytes communicate through a canalicular system that maintains the vitality and mineral metabolism of bone. Casting the vascular canals and canaliculi of compact bone with methacrylate and viewing them with scanning electron microscopy shows their extent and relationships. Confocal laser scanning microscopy of the same specimen before corrosion establishes the degree of calcification of the different tissue components. These methods were used to compare basal with alveolar compact bone in the rat mandible at different ages. Sections of the mandibular molar region were placed in a methacrylate resin. After polymerization and study with confocal microscopy, the organic matrix was removed. Juvenile rats had large irregular central vascular canals and lacunae that were more concentric in the basal than the alveolar bone. Cast lacunae were round, and the canaliculi from these lacunae were short and thick in both bones. Adult rats had regular concentrically arranged lacunae in the basal bone. Cast lacunae were ellipsoid and flatter in the basal bone than in the alveolar bone. The intercommunicating canaliculi were increased and canaliculi had more branching than the juvenile rats. The aged rats had fewer vascular canals, lacunae, and canaliculi and had osteoporotic changes. The cast lacunae were slender and flat especially in the basal bone. The porosity of the mandible became more pronounced in the alveolar than in the basal bone with aging. The canaliculi of mandibular compact bone thinned and developed extensive branching with adulthood but decreased in size and number with advanced age. Lacunae proceed from the large circular structures of youth to the flat forms of the aged. These studies show that the internal structure of compact bone changes with age and mirrors its functional state.

Acute effects of ovariectomy on wound healing of alveolar bone after maxillary molar extraction in aged rats.

Acute effects of ovariectomy on the bone wound healing processes after maxillary molar extraction in aged rats were examined by means of quantitative scanning electron microscopy (SEM), backscattered electron image (BSE) analysis and energy-dispersive X-ray (EDX) microanalysis. Six-month-old female rats underwent either sham operation or bilateral ovariectomy, and 7 days postoperatively, the maxillary first molars were extracted. On post-extraction days 7, 30 and 60, the dissected maxillary bone surfaces were examined by SEM to reveal the bone formative and resorptive areas around the extracted alveolar sockets. In addition, the resin-embedded maxillae were micromilled in the transverse direction through the extracted alveolar sockets, and the newly-formed bone mass on the buccal bone surfaces and within the extracted sockets was examined by BSE analysis. Compared with sham-operated controls, the extent of newly-formed bone mass on the buccal bone surfaces in OVX rats was significantly decreased, due to increased bone resorption. On the other hand, new bone formation within the extracted sockets was similar in the experimental groups. In EDX microanalysis of these newly-formed bone matrices, both Ca and P weight % and Ca/P molar ratio were similar in the experimental groups. Our results suggest that 1) acute estrogen deficiency induced by ovariectomy stimulates sustained bone resorption, but has less effect on bone formation, and 2) bone wound healing after maxillary molar extraction within extracted alveolar sockets is not significantly delayed by ovariectomy, but bony support by newly-formed bone mass on the maxillary bone surfaces at the buccal side of the extracted sockets is significantly decreased, due to increased bone resorption.

Bone wound healing after maxillary molar extraction in ovariectomized aged rats: quantitative backscattered electron image analysis.

The processes of bone wound healing after maxillary molar extraction in ovariectomized aged rats were examined by means of quantitative backscattered electron image analysis and energy-dispersive X-ray microanalysis. Six-month-old female rats were either sham-operated or underwent bilateral ovariectomy (OVX), and 60 days postoperatively, the maxillary first molars were extracted. On post-extraction days 7, 30, and 60, the dissected and resin-embedded maxillae were micromilled in the transverse direction through the extracted alveolar sockets, and new bone formation on the buccal maxillary bone surface and within the extracted alveolar sockets was examined. In both sham-operated control and OVX rats, new bone formation was recognized on the buccal bone surface, as well as within the extracted sockets, and increased daily through to day 60. In comparison to sham-operated controls, new bone formation in OVX rats was significantly decreased both on the buccal bone surface and within the extracted sockets. Our results suggest that bone wound healing by new bone formation after maxillary molar extraction is significantly decreased in OVX-induced osteoporosis.