Ultrastructural study by backscattered electron imaging and elemental microanalysis of bone-to-biomaterial interface and mineral degradation of porcine xenografts used in maxillary sinus floor elevation.

OBJECTIVES: The aim of this study was to carry out an ultrastructural study of the biomaterial-to-bone interface and biomaterial mineral degradation in retrieved bone biopsies following maxillary sinus augmentation using collagenized porcine xenografts (Osteobiol(®) Mp3) in 15 clinical cases. MATERIALS AND METHODS: Nine months after sinus lifting, bone cores were harvested from the maxillary sinus. The specimens were processed for observation under a scanning electron microscope with backscattered electron imaging (SEM-BSE). In addition, chemical analysis and elemental mapping of the mineral composition were generated using a microanalytical system based on energy-dispersive X-ray spectrometry (EDX). RESULTS: No clinical complications were evident during surgery. Scanning electron microscopy revealed that newly formed bone had become closely attached to the xenograft. Statistical analysis showed a significantly high Ca/P ratio in the biomaterial (2.46 ± 0.16) and at the bone interface (2.00 ± 0.48) compared to bone (1.97 ± 0.36), which suggests that there may be a gradual diffusion of Ca ions from the biomaterial into the newly forming bone at the interface as part of the biomaterial's resorption process. EDX analysis of the residual porcine biomaterial at different points showed some particle categories with different mean ratios of Ca/P according to size, pointing to different stages of the resorption process. CONCLUSIONS: The biomaterial proved to be biocompatible, bioreabsorbable and osteoconductive when used as a bone substitute for maxillary sinus elevation. SEM-BSE revealed that newly formed bone had become closely attached to the xenografts. EDX analysis monitored the resorption process of the porcine bone xenograft. Elemental mapping showed that there was a gradual diffusion of Ca ions from the biomaterial to the newly forming bone at the interface.

Ultrastructural study by backscattered electron imaging and elemental microanalysis of biomaterial-to-bone interface and mineral degradation of bovine xenografts in maxillary sinus floor elevation.

OBJECTIVES: The aim of this study was to perform an ultrastructural study of the bone-to-biomaterial interface and biomaterial mineral degradation in retrieved bone biopsies following maxillary sinus augmentation using bovine xenografts (Endobon(®)) in 15 clinical cases. MATERIALS AND METHODS: Nine months after sinus lifting, bone cores were harvested from the maxillary sinus. The specimens were processed for observation under a scanning electron microscope with backscattered electron imaging (SEM-BSE). In addition, chemical analysis and elemental mapping of the mineral composition were generated using a microanalytical system based on energy-dispersive X-ray spectrometry (EDX). RESULTS: No clinical complications occurred during surgery. Scanning electron microscopy revealed that newly formed bone was closely attached to the xenograft. Elemental analysis showed a significantly high Ca/P ratio in the residual biomaterials (3.031 ± 0.104) compared with the interface (2.908 ± 0.115) and new bone (2.889 ± 0.113), which suggests that there may be a gradual diffusion of Ca ions from the biomaterial into the newly forming bone at the interface as part of the biomaterial's resorption process. EDX analysis of the residual bovine biomaterial showed particle categories with different mean Ca/P ratios according to size, pointing to different stages of the resorption process. CONCLUSIONS: The biomaterial proved to be biocompatible and osteoconductive when used as a bone substitute for maxillary sinus elevation. The grafting material used is not a completely resorbable material over the time period covered by this study.

[Structure of maxillary sinus mucous membrane under normal conditions and in odontogenic perforative sinusitis].

Methods of light, electron microscopy and immunohistochemistry were used to study the samples of maxillary sinus (MS) mucous membrane (MM) under normal conditions and in odontogenic sinusitis. To study the normal structure, the samples were obtained at autopsy from 26 human corpses 12-24 hours after death. Electron microscopic and immunohistochemical study was performed on biopsies of grossly morphologically unchanged MS MM, obtained during the operations for retention cysts in 6 patients. MS MM in perforative sinusitis was studied using the biopsies obtained from 43 patients. The material is broken into 4 groups depending on perforative sinusitis duration. Under normal conditions, MS MM is lined with a pseudostratified columnar ciliated epithelium. Degenerative changes of ciliated epithelial cells were already detected at short time intervals after MS perforations and become apparent due to reduction of specific volume of mitochondria and, rough endoplasmic reticulum, and increase of nuclear-cytoplasmic ratio. In the globlet cells, the reduction of nuclear-cytoplasmic ratio was associated with the disturbance of the secretory product release. At time intervals exceeding 3 months, epithelium underwent metaplasia into simple cuboidal and stratified squamous keratinized, while in MS MM lamina propria, cellular infiltration was increased. CD4+ cell content in sinus MM gradually increased, while at late periods after perforation occurrence it decreased. Low CD4+ cell count within the epithelium and the absence of muromidase on the surface of MS MM was detected. With the increase of the time interval since MS perforation, the number of CD8+ and CD20+ cells in MS MM was found to increase.

Back-scattered electron imaging and elemental microanalysis of retrieved bone tissue following maxillary sinus floor augmentation with calcium sulphate.

OBJECTIVES: To investigate the presence and composition of residual bone graft substitute material in bone biopsies from the maxillary sinus of human subjects, following augmentation with calcium sulphate (CaS). MATERIAL AND METHODS: Bone cores were harvested from the maxillary sinus of patients who had undergone a sinus lift procedure using CaS G170 granules 4 months after the initial surgery. Samples from seven patients, which contained residual biomaterial particles, were examined by field emission scanning electron microscopy and energy dispersive X-ray spectroscopy was used to determine the composition of the remaining bone graft substitute material. RESULTS: Residual graft material occurred in isolated areas surrounded by bone and consisted of individual particles up to 1 mm in length and smaller spherical granules. On the basis of 187 separate point analyses, the residual material was divided into three categories (A, B and C) consisting of: A, mainly CaS (S/P atomic% ratio > or =2.41); B, a heterogeneous mixture of CaS and calcium phosphate (S/P=0.11-2.4) and C, mainly calcium phosphate (S/P< or =0.11; C), which had a mean Ca : P ratio of 1.63+/-0.2, consistent with Ca-deficient hydroxyapatite. Linescans and elemental maps showed that type C material was present in areas which appeared dense and surrounded, or were adjacent to, more granular CaS-containing material, and also occurred as spherical particles. The latter could be disintegrating calcium phosphate in the final stages of the resorption process. CONCLUSIONS: CaS resorption in the human maxillary sinus is accompanied by CaP precipitation which may contribute to its biocompatibility and rapid replacement by bone.

A histologic and histomorphometric evaluation of anorganic bovine bone retrieved 9 years after a sinus augmentation procedure.

BACKGROUND: Anorganic bovine bone (ABB) has been shown to have osteoconductive properties and no inflammatory or adverse responses as grafting materials used in sinus augmentation procedures. Despite these successful results, histologic data in humans over the long-term period are scarce. The purpose of this study was to analyze the histomorphometric data 9 years after surgery in a case of maxillary sinus augmentation using ABB. METHODS: The histologic evaluation was performed in five different thin sections of the specimen, comparing histomorphometric measures for newly formed bone, marrow spaces, biomaterial particles remnants, and number of osteocytes embedded in both trabecular bone and bone tissue near the ABB. The investigation was carried out by means of scanning electron microscopy and brightfield and circularly polarized light microscopy. RESULTS: We observed a mean amount of newly formed bone of 46.0% +/- 4.67%, ABB remnants of 16.0% +/- 5.89%, and marrow spaces of 38.0% +/- 8.93%. The osteocyte index was 4.43 for bone around ABB and 3.27 in the trabecular bone at a distance from the particles. CONCLUSIONS: After 9 years, the tissue pattern appeared composed by residual ABB particles in close contact to the newly formed bone. The bone mineralized matrix around the ABB had collagen fibers randomly oriented and more osteocytes embedded. The results demonstrate both a high level of osteoconductivity and a "biomimetic" behavior over the long term.

The presence of bacterial microcolonies on the maxillary sinus ciliary epithelium in healthy young individuals.

OBJECTIVE: The aim of this cross-sectional in vitro study was to evaluate the mucosal surfaces of healthy maxillary sinuses, explore different forms of bacterial microorganism colonies present on the mucous membrane, and determine a mucosal surface area they occupy. METHODS: Samples of the maxillary sinus mucosa were collected from 30 healthy patients (M = 11; F = 19). The material was obtained during the Le Fort I osteotomy performed during corrective jaw surgery. The morphological and morphometric analysis of sinus mucosa and bacterial film that was grown on it was performed using scanning electron microscopy (SEM) as well as imaging software. RESULTS: Scanning electron microscopy analysis showed the presence of different bacterium and bacteria-like structures in all the analyzed samples. In most cases, the bacterial film was mostly composed of diplococci-like and streptococci-like structures on the mucosa of the paranasal sinus. In any case, the mucous layer did not cover the whole lining of the evaluated sample. Each colony consists of more than 20 single bacterial cells, which has grown in aggregates. CONCLUSIONS: Under the conditions of normal homeostasis of the body, the maxillary sinuses present diverse bacterial colonization. The bacteria are dispersed or concentrated in single microcolonies of the biofilm on the border of the mucous covering the ciliary epithelium. There is no uniform layer of the biofilm covering the mucosa of the maxillary sinuses. Because the biofilm is detected on healthy individuals sinus mucosa, the clinical question if it may become pathogenic is unclear and require an explanation.

Histologic and ultrastructural analysis of regenerated bone in maxillary sinus augmentation using a porcine bone-derived biomaterial.

BACKGROUND: The purpose of the present study was the histologic and ultrastructural evaluation of a biomaterial composed of cortical pig bone in the form of granules. METHODS: After maxillary sinus augmentation using this biomaterial, 10 specimens were retrieved after 5 months in 10 patients using this biomaterial. The specimens were processed to be observed under light microscopy (LM) and transmission electron microscopy (TEM). Histomorphometric measurements were presented by means +/- standard deviations. RESULTS: LM showed that most of the particles were surrounded by newly formed bone. In some areas, the osteoid matrix was present; however, mainly compact bone was present at the interface. There was no evidence of an acute inflammatory infiltrate. The newly formed bone was 36% +/- 2.8% and marrow spaces were 38% +/- 1.6%, whereas residual grafted material was 31% +/- 1.6%. Under TEM, all phases of bone formation (osteoid matrix, woven, and lamellar bone) were observed in proximity with the biomaterial particles. The bone-biomaterial interface showed a close contact between the porcine bone particles and the surrounding bone that had mainly features of mature bone with numerous osteocytes. A lamina limitans was sometimes present at this interface. CONCLUSIONS: According to our knowledge, this is the first study presenting data on TEM of a porcine bone-derived biomaterial used in sinus augmentation procedures in humans. Our findings show that this is a biocompatible biomaterial that can be used for maxillary sinus augmentation procedures without interfering with the normal reparative bone processes.

[An increase of plasma cells in the regenerated mucous membrane of maxillary sinus. Experimental study]. / Aumento de células plasmáticas en la mucosa regenerada del seno maxilar. Estudio experimental.

In ten albino New Zealand rabbits we studied in the regenerated mucosa of the maxillary sinus what happens after three months of radical removal of sinus mucosa. The study was realized by light and electronic microscopy, with special attention to the plasma cells around the submucosal glands. There is an increased of plasma cells, in a proportion of 10:1.

[Basal and non-hair cells in the regenerated mucosa of the maxillary sinus]. / Células basales y no ciliadas en la mucosa regenerada del seno maxilar. Estudio experimental.

In 10 albino New Zealand rabbits, a study was made of the regeneration of the hair cells in the maxillary sinus mucosa three months after removing the sinus mucosa. Using optical microscopy and transmission and scanning electron microscopy, basal and non-hair cells were studied as the probable origin of hair cells. The regenerated sinus mucosa showed areas of epithelium consisting only of a basement membrane and a layer of basal cells. Ultrastructurally, two types of basal cell were visible in the regenerated mucosa. The proportion of non-hair cells was greater than in normal mucosa.

[Lower antrostomy in surgery of the maxillary sinus: experimental study]. / La antrostomía inferior en la cirugía del seno maxilar: estudio experimental.

Inferior nasoantral windows, with or without radical surgical removal of the maxillary sinus mucosa, were evaluated in 15 New Zealand white rabbits. After three months, specimens were obtained for examination. Bacteriological cultures and light and electron microscopic studies were made. Mucociliary clearance was studied. Differences were found in the antrostomy patency rate between radical antrostomy and inferior nasoantral windows. After three months, the creation of a temporary inferior nasoantral window did not change the maxillary sinus mucosa and did not increase the rate of sinus infections in rabbits.

Experimental study on histological changes in the sinus membrane following sinus lift.

UNLABELLED: The aim of this study is to assess the histological changes in the sinus mucosa adjacent to the alloplastic material used for subantral augmentation. MATERIALS AND METHODS: The study included ten sheep and a dog. The first group of five sheep underwent a sinus lift procedure, using PerioGlas as an augmentation material; the second similar group of sheep was the control group. The dog underwent a sinus lift procedure, with PerioGlas augmentation, after the sinus membrane was intentionally perforated and two implants were placed in the same operative step. RESULTS: Scanning electron microscopy (SEM) of the sinus mucosa in the control group revealed cells without cilia between goblet cells. The cilia were uniformly arranged in sections in the same direction. Changes occurred in the sinus mucosa after grafting, such as drastic reduction of ciliated cells, which seemed to be replaced by goblet cells. In all sheep undergoing grafting, generalized fibrosis was found in the mucosal area that came into contact with PerioGlas. In two of the sheep in which grafting was performed, mucoid cysts with pseudo stratified ciliated epithelium were present. Even when the sinus mucosa was perforated (in the dog), the inflammatory process developed in the mucosa did not prevent the integration of the graft and implants. In conclusion, following the sinus lift procedure, changes occur in the sinus membrane to adapt to the new situation, without the appearance of chronic or acute suppurative processes.

Histologic changes in the maxillary sinus membrane after sinus membrane elevation and the simultaneous insertion of dental implants without the use of grafting materials.

OBJECTIVE: The aim of this study was to evaluate the histologic changes in the maxillary sinus membrane after sinus membrane elevation and the simultaneous insertion of dental implants without additional grafting material. STUDY DESIGN: In 6 mongrel dogs, bilateral edentulated flat alveolar ridges were created in the maxilla. After 3 months of healing, an implant was unilaterally placed in the maxillary sinus in such a way that it protruded 5 mm into the maxillary sinus after sinus membrane elevation. On the opposite side, the maxillary sinus was left untreated as a control site. The animals were killed 6 months after surgery. The maxillary sinus mucosa was examined using light microscopy and scanning and transmission electronic microscopy. RESULTS: There were no morphologic or ultrastructural differences in the sinus membrane between groups. CONCLUSION: The results indicate that the surgical procedure by which implants are inserted into the sinus cavity by elevating the sinus membrane without adding any graft material appears to have little influence on the histologic characteristics of the sinus membrane.

Human histologic and electromicroscopic analysis with synthetic peptide enhanced hydroxyapatite in the maxillary sinus elevation procedure: a case report.

Elevation of the sinus floor to augment the alveolar process to place implants is an effective treatment. The traditional use of autologous bone implies a secondary donor site and the related morbidity, and, hence, is not readily accepted by many patients. The new graft material, synthetic peptide enhanced hydroxyapatite (PepGen P-15 Flow, Ceramed Dental, Lakewood, CO, DENTSPLY Int.,York, PA), shows the ease in use and ability to generate new bone growth in the sinus elevation procedure. Histologic and electromicroscopic analysis was performed on the new bone formed after the application.

Maxillary sinus augmentation using different grafting materials and osseointegrated dental implants in monkeys. Part II. Evaluation of porous hydroxyapatite as a grafting material.

The aim of this study was to evaluate clinically, histologically and histometrically the use of porous hydroxyapatite (i.e. Interpore-200) as a bone grafting material for maxillary sinus augmentation procedures. In 4 adult male rhesus monkeys (i.e. Macaca mulatta) the 1st, 2nd and 3rd maxillary molars on one side of the jaws were extracted and the remaining bone between the alveolar crest and the bottom of the sinus was reduced to 3-4 mm. After 3 months, maxillary sinus augmentation procedures were performed on one side of the jaws in each monkey, and the sinuses grafted with the porous hydroxyapatite. Two IMZ titanium plasma-sprayed cylinder implants were then immediately placed into the augmented sinus (i.e. simultaneous implants-loaded group). After 4 months, 2 additional similar implants were placed into the previously augmented sinuses (i.e. delayed implants-loaded group). Four months later, the abutment connection was performed and all 4 implants were loaded with a gold-alloy bridge for 6 months (i.e. until sacrifice). The contralateral side of each monkey received the same treatment with the exception that the extractions were performed 7 months after those in the opposite side and that the implants in this side were not loaded. Thus, 2 additional study groups (i.e. simultaneous implants-unloaded group and delayed implants-unloaded group) were obtained. Clinically, all loaded implants were stable at the day of sacrifice. Histologic analysis demonstrated a significant amount of new bone formation in the augmented sinuses. Porous hydroxyapatite graft particles appeared to be integrated to the new bone. The percentage of direct mineralized bone-to-implant contact in the augmented area was greater on the delayed-placed implants than on the simultaneously-placed implants. Also, the percentage of direct mineralized bone-to-implant contact was greater in the residual bone than in the augmented area. It was concluded that this porous hydroxyapatite bone graft enhanced bone formation and bone-to-implant contact in the augmented sinuses and that the delayed implant placement in combination with the sinus augmentation procedure appears to result in a higher percentage of direct mineralized bone-to-implant contact.

Maxillary sinus augmentation using different grafting materials and dental implants in monkeys. Part IV. Evaluation of hydroxyapatite-coated implants.

The aim of this study was to evaluate clinically, histologically and histometrically the use of hydroxyapatite-coated dental implants in conjunction with maxillary sinus augmentation procedures. In 4 adult male Rhesus monkeys (Macaca mulatta) the 3 maxillary molars on 1 side of the jaws were extracted and the remaining bone between the alveolar crest and the floor of the sinus was reduced to 3-4 mm. After 3 months, maxillary sinus augmentation procedures were performed in each monkey and the sinuses were grafted with a porous hydroxyapatite bone graft (Interpore-200). At the same time, 2 hydroxyapatite-coated cylinder implants (IMZ) were immediately placed into the augmented sinuses (i.e. simultaneous-implants-loaded group). Four months later, 2 additional similar implants were placed into the previously augmented sinuses (i.e. delayed-implants-loaded group). After 4 months, the abutment connection was performed and all 4 implants were loaded with a gold-alloy bridge for 6 months (i.e. until sacrifice of the animals). The contralateral side of each monkey received the same treatment with the exception that removal of the maxillary molars was performed 7 months after those in the opposite side, and that the implants in this side were not loaded. Thus, 2 additional study groups (i.e. simultaneous-implants-unloaded group and delayed-implants-unloaded group) were obtained. Clinically, all loaded and unloaded implants were stable the day of sacrifice. Histologically, the grafted sinuses exhibited a significant amount of new bone formation with integration of the porous hydroxyapatite graft particles and hydroxyapatite-coat of the dental implants to the new bone. Histometric analysis indicated that on the loaded side the implants placed simultaneously with the sinus lift procedure exhibited greater direct mineralized bone-to-implant contact than the delayed placed implants. In addition, the percentage of direct mineralized bone-to-implant contact was significantly greater in the residual bone in comparison to the augmented area in all groups. Loading of the implants exhibited a positive effect on the percentage of direct mineralized bone-to-implant contact in the augmented area. It could be concluded that hydroxyapatite-coated implants may be of benefit when used in conjunction with sinus augmentation procedures.

Maxillary sinus augmentation using different grafting materials and dental implants in monkeys. Part I. Evaluation of anorganic bovine-derived bone matrix.

The aim of this study was to evaluate clinically, histologically and histometrically the use of anorganic bovine bone matrix (i.e. Bio-oss) as a grafting material for maxillary sinus augmentation procedures. In 4 adult male rhesus monkeys (i.e. Macaca mulatta) the 1st, 2nd and 3rd maxillary molars on one side of the jaws were extracted. The remaining bone between the alveolar crest and the bottom of the sinus was then reduced to 3-4 mm. After 3 months, maxillary sinus augmentation procedures were performed on one side of the jaws in each monkey and the sinuses were grafted with the bovine bone matrix. At that time, 2 IMZ pure titanium plasma coated implants were immediately placed into the augmented sinuses (i.e. simultaneous implants-loaded group). After 4 months, 2 additional similar implants were placed into these previously augmented sinuses (i.e. delayed implants-loaded group). Four months later, the abutment connection was performed and all 4 implants were loaded with a gold-alloy bridge for 6 months (i.e. until sacrifice of the animals). The contralateral side of each monkey received the same treatment with the exception that the extractions were performed 7 months after those in the opposite side and that the implants in this side were not loaded. Thus, 2 additional study groups (i.e. simultaneous implants-unloaded group and delayed implants-unloaded group) were obtained. Clinically, all loaded implants were stable at the day of sacrifice. Histologically, the grafted sinuses exhibited significant bone formation with integration of the bovine bone matrix particles to the new bone. Direct mineralized bone-to-implant contact was greater for the delayed implant placement groups than for the implants installed simultaneously with the sinus augmentation. Furthermore, the percentage of direct mineralized bone-to-implant contact was greater in the residual bone than in the augmented area. It was concluded that the anorganic bovine bone matrix facilitated bone formation and implant osseointegration in the augmented sinuses and that the delayed implant placement in combination with the sinus augmentation procedure seemed to be preferable.