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

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

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

Comparison of bone graft healing between autogenous bone, blood clot and anorganic bovine bone matrix: radiographic and histological analyses / Comparasión de la reparación de injertos óseos entre hueso autógeno, coágulo sanguíneo y matriz ósea bovina anorgánica: análisis histológico y radiográfico

Surgical procedures involving the rehabilitation of the maxillofacial region frequently require use of bone grafts. Bone graft follow-up is generally done through clinical analysis and especially through image tests, although few studies specifically correlate both. The object of this research was to establish a relation between radiographic exams and the bone repair process stage in created defect with autogenous bone, blood clot and anorganic bovine bone matrix. Three 8 mm diameter defects were performed in the parietal bone of 6 male adult beagle dogs, choosing the selected graft for each defects; 3 and 6 week period were used for radiographic and histological analyses. The result show that autogenous bone and blood clot were similar between histological and radiograph analyses; for heterogeneous bone was present areas described how bone in radiograph that were residual particles in histological exam. We concluded that radiographic tests could be used as a parameter for reconstruction follow-up only when autogenous bone graft is used.

Los procedimientos que envuelven la rehabilitación de la región maxilofacial frecuentemente requieren el uso de injertos óseos. El seguimiento de la evolución del injerto óseo frecuentemente es realizado con análisis clínico y especialmente a través de estudios de imágenes, aunque pocos estudios han correlacionado ambos. El objetivo de esta investigación fue establecer la relación entre radiografías y las etapas de la reparación ósea en defectos creados con hueso autógeno, coagulo sanguíneo y matriz ósea de hueso bovino. Tres defectos de 8 mm de diámetro fueron realizados en el parietal de 6 perros adultos, escogiendo el injerto seleccionado para cada defecto; 3 y 6 semanas después fueron realizados los estudios histológicos y radiográficos. Los resultados mostraron que el hueso autógeno y el coágulo sanguíneo presentaron semejanzas en los análisis histológico y radiográfico; para el hueso bovino se observó áreas decritas como hueso en la radiografia mientras realmente fueron partículas de hueso heterogeno descritas en el análisis histológico. Concluimos que el test radiográfico puede ser usado como parámetro para la reconstrucción y seguimiento preferentemente cuando es usado el injerto autógeno.

Regeneración ósea en defectos críticos tratados con matriz ósea bovina inorgánica aplicada con dos diferentes vehículos / Bone regeneration in critical defect treated with bovine inorganic bone matrix with two different carriers

Múltiples materiales se han utilizado en la reconstrucción ósea; la matriz ósea bovina ha sido ampliamente estudiada y aun existen investigaciones que identifican su capacidad de contribuir en la formación ósea. El objetivo de esta investigación fue establecer la reparación ósea en defectos críticos de calotas de perros rellenados con matriz ósea bovina inorgánica transportada con carboxi-metil-celulosa. Se diseñó un estudio experimental en 6 perros donde se realizaron 3 defectos de 8mm de diámetro en los parietales de cada animal; el defecto fue rellenado aleatoriamente con hueso autógeno particulado (grupo I), matriz ósea bovina inorgánica con suero fisiológico al 0,9 por ciento (grupo II) y matriz ósea bovina con carboxi-metil-celulosa (grupo III). Se realizó el sacrificio de los animales a la tercera y sexta semana momento en el que se realizaron los estudios radiográficos de los defectos tratados y los estudios histológicos de rutina con tinciones de hematoxilina-eosina. No se observó procesos infecciosos vinculados a los materiales injertados. En el grupo del hueso autógeno se observó una adecuada aposición ósea en la tercera y sexta semana junto a una relación correcta con la imagen radiográfica; en el grupo II se observó etapas de formación ósea y permanencia de los fragmentos de hueso injertado; en el grupo III se observó infiltrado inflamatorio en los dos periodos de sacrificio, abundante tejido conectivo y bajo nivel de formación ósea. Las radiografías de los grupos II y III mostraron signos de formación ósea que en la histología eran sólo remanentes del material injertado. Se concluye que la matriz ósea bovina inorgánica es compatible con el tejido óseo y que puede contribuir a la formación ósea aunque el empleo de la carboxi-metil-celulosa, como vehículo, podría obstaculizar la regeneración ósea.

In bone reconstruction has been used different bioamterials; bovine bone matrix has been studied and nowadays new research analyses the capacity for bone regeneration. The aim of this research was to evaluate the bone reparation in critical defects on skull of dog filled with inorganic bovine bone matrix carried by carboxi-methyl-celulose. Was design an experimental research with 6 dog; on parietal bone was realized a 8mm diameter defect with a trephine and the defect was filled by particle autogenous bone (group I), inorganic bovine bone matrix carried with saline solution (0,9 percent) (group II) and inorganic bovine bone matrix carried with carboxi-methyl-celulose. Was realized the sacrifice of animals in a third and sixth week and was performed the radiographic image and the histological study with hemaotoxilin-eusin in a routine technique. Non infection was observed in any of sample. In the autogenous bone graft was observed an adequately bone formation in the third and sixth week analyses and was related to radiographic image; for group II was observed some bone formation and presence of bovine bone particles and for group III was observed inflammatory cells for two period of analyses with a low level of bone formation. The radiographic analyses show sign of bone formation but histological analyses show only permanence of bovine bone particles confounding the radiograph results. Is conclude that bovine bone inorganic matrix in COMPATIBLE con bone tissue and can contributed to bone formation although the use of carboxi-methyl-celulose can be an obstacle for bone regeneration.

Treatment of Intrabony Defects with Anorganic Bone Matrix/P-15 or Guided Tissue Regeneration in Patients with Aggressive Periodontitis

Intrabony periodontal defects present a particular treatment problem, especially in patients with generalized aggressive periodontitis (G-AgP). Regenerative procedures have been indicated for this clinical situation. The aim of this study was to compare treatment outcomes of intrabony periodontal defects with either anorganic bone matrix/cell binding peptide (ABM/P-15) or guided tissue regeneration (GTR) in patients with G-AgP. Fifteen patients, with two intrabony defects ≥3 mm deep, were selected. Patients were randomly allocated to be treated with ABM/P-15 or GTR. At baseline and at 3 and 6 months after surgery, clinical and radiographic parameters and IL-1β and IL-6 gingival fluid concentrations were recorded. There was a significant probing pocket depth reduction (p<0.001) for both groups (2.27 ± 0.96 mm for ABM/P-15 group and 2.57 ± 1.06 mm for GTR group). Clinical attachment level gain (1.87 ± 0.94 mm for ABM/P-15 group and 2.09 ± 0.88 mm for GTR group) was also observed. There were no statistically significant differences in clinical parameters between the groups. The radiographic bone fill was more expressive in ABM/P-15 group (2.49 mm) than in GTR group (0.73 mm). In subtraction radiographs, the areas representing gain in density were 93.16% of the baseline defect for ABM/P-15 group versus 62.03% in GRT group. There were no statistically significant differences in inter-group and intra-group comparisons with regards to IL-1β and IL-6 quantification. Treatment of intrabony periodontal defects in patients with G-AgP with ABM/P-15 and GTR improved significantly the clinical outcomes. The use of ABM/P-15 promoted a better radiographic bone fill.

Defeitos periodontais infra-ósseos representam um desafio particular no tratamento, especialmente em pacientes com periodontite agressiva generalizada (PAg-G). Procedimentos regenerativos tem sido indicados para esta situação clínica. O objetivo deste estudo foi comparar os resultados do tratamento de defeitos periodontais infra-ósseos com associação de matriz óssea inorgânica bovina com o P-15 (MOI/P-15) ou regeneração tecidual guiada (RTG) em pacientes com PAg-G. 15 pacientes com PAg-G, com pelo menos dois defeitos periodontais infra-ósseos (profundidade de sondagem ≥4 mm e componente infra-ósseo ≥3 mm) foram selecionados. Os pacientes foram aleatoriamente alocados para serem tratados com MOI/P-15 ou RTG. No exame inicial, e aos 3 e 6 meses após a cirurgia, os parâmetros clínicos e radiográficos e as concentrações de IL-1β e IL-6 no fluido gengival foram registrados. Houve uma redução significativa profundidade de sondagem (p<0,001) para ambos os grupos (2,27 ± 0,96 mm para o grupo MOI/P-15 e 2,57 ± 1,06 mm para o grupo RTG). Um ganho no nível clínico de inserção (1,87 ± 0,94 mm para o grupo MOI/P-15 e 2,09 ± 0,88 mm para o grupo RTG) também foi observado. Na comparação entre grupos, não houve diferenças estatisticamente significativas nos parâmetros clínicos. O preenchimento ósseo radiográfico foi mais expressivo no grupo MOI/P-15 (2,49 mm) do que no grupo RTG (0,73 mm). Na análise radiográfica, as radiografias de subtração apresentaram ganho médio de área radiopaca em relação ao defeito inicial de 93,16% para grupo MOI/P-15, contra 62,03% para o grupo RTG. Na análise das citocinas, não foram observadas diferenças estatisticamente significantes nas comparações intra e entre os grupos. O tratamento de defeitos infra-ósseos com MOI/P-15 ou RTG em pacientes com PAg-G, em um período de 6 meses, levou a melhoras nos parâmetros clínicos. O uso de MOI/P-15 levou a um maior preenchimento radiográfico.

Grafting Using Injectable Calcium Sulfate in Bone Tumor Surgery: Comparison with Demineralized Bone Matrix-based Grafting

BACKGROUND: Injectable calcium sulfate is a clinically proven osteoconductive biomaterial, and it is an injectable, resorbable and semi-structural bone graft material. The purpose of this study was to validate the clinical outcomes of injectable calcium sulfate (ICS) grafts as compared with those of a demineralized bone matrix (DBM)-based graft for filling in contained bony defects created by tumor surgery. METHODS: Fifty-six patients (41 males and 15 females) with various bone tumors and who were surgically treated between September 2003 and October 2007 were included for this study. The patients were randomly allocated into two groups, and either an ICS graft (28 patients) or a DBM-based graft (28 patients) was implanted into each contained defect that was developed by the surgery. The radiographic outcomes were compared between the two groups and various clinical factors were included for the statistical analysis. RESULTS: When one case with early postoperative pathologic fracture in the DBM group was excluded, the overall success rates of the ICS and DBM grafting were 85.7% (24/28) and 88.9% (24/27) (p > 0.05), respectively. The average time to complete healing was 17.3 weeks in the ICS group and 14.9 weeks in the DBM group (p > 0.05). Additionally, the ICS was completely resorbed within 3 months, except for one case. CONCLUSIONS: Although the rate of resorption of ICS is a concern, the injectable calcium sulfate appears to be a comparable bone graft substitute for a DBM-based graft, with a lower cost, for the treatment of the bone defects created during surgery for various bone tumors.

Radiographic follow-up evaluation of sinus augmentation with deproteinized bovine bone and implant installation after loading.

Computed tomography (CT) has been used in diagnosis and implant treatment, and CT can be used to assess the dimensional stability of graft materials after maxillary sinus augmentation. A 50-year-old male patient was treated for the simultaneous placement of implants with sinus augmentation and two post-operative CT scans were performed after the delivery of the prosthesis at 9 months and 15 months after the operation. There were no significant changes in alveolar bone height and the buccal window seemed to show remodeling over time. The continuity of the defect and the cortication were apparent in the 15-month post-operative CT scan. Implants installed simultaneously with sinus augmentation were well in function and the graft material seemed to be stable in maintaining tissue dimensions after the loading of the implants. The presented results need to be validated in further large case series or case-controlled studies.

Comparison of the regenerative potential of an allograft used alone and that in conjunction with an immunosuppressive drug in the treatment of human periodontal infrabony defects: A clinical and radiological study.

Background: Experimentation confirmed the conclusion that bone allografts, like other tissue and organ allografts, are immunogenic. These immune responses cause resorption of allograft, thus lowering the bone formation capacity of the graft. An attempt has been made in this study to prevent immune reactions and achieve enhanced regeneration of allograft-demineralized freeze-dried bone matrix by incorporating it with an immunosuppressive drug Cyclosporine-A (CsA) in the treatment of human periodontal infrabony defects. Materials and Methods: Fifteen patients showing clinical evidence of almost bilateral infrabony defects requiring bone grafting procedures were selected. In each patient, the infrabony defect of one side of the arch was designated as Group A (control site) and the infrabony defect of the contralateral side of the same arch was designated as Group B (test site). Results: On comparative evaluation of the two groups (by Student t-test), the mean values of reduction in probing depth (P=0.81 NS ) and gain in clinical attachment level (P=1.00 NS ) of Group B were found to be greater than that of Group-A, but the differences were statistically non-significant. The mean linear bone fill (P=0.010 ** ) of Group B was also detected to be higher than that of Group A, and the difference was found to be statistically significant. Conclusion: Increase in linear bone fill in Group B signifies the role of CsA in augmenting the regenerative potential of allograft by eliminating immune reactions.

Digital subtraction radiographic analysis of the combination of bioabsorbable membrane and bovine morphogenetic protein pool in human periodontal infrabony defects

OBJECTIVES: This study assessed the bone density gain and its relationship with the periodontal clinical parameters in a case series of a regenerative therapy procedure. MATERIAL AND METHODS: Using a split-mouth study design, 10 pairs of infrabony defects from 15 patients were treated with a pool of bovine bone morphogenetic proteins associated with collagen membrane (test sites) or collagen membrane only (control sites). The periodontal healing was clinically and radiographically monitored for six months. Standardized pre-surgical and 6-month postoperative radiographs were digitized for digital subtraction analysis, which showed relative bone density gain in both groups of 0.034 ± 0.423 and 0.105 ± 0.423 in the test and control group, respectively (p>0.05). RESULTS: As regards the area size of bone density change, the influence of the therapy was detected in 2.5 mm² in the test group and 2 mm² in the control group (p>0.05). Additionally, no correlation was observed between the favorable clinical results and the bone density gain measured by digital subtraction radiography (p>0.05). CONCLUSIONS: The findings of this study suggest that the clinical benefit of the regenerative therapy observed did not come with significant bone density gains. Long-term evaluation may lead to a different conclusions.

Restoration of the maxillary anterior tooth using immediate implantation with simultaneous ridge augmentation.

Atrophy is most severe during the first month of post-extraction in the anterior maxilla with the degree of horizontal bone resorption being nearly twice as high as that of vertical bone resorption. The loss of the buccal alveolar plate following tooth extraction may lead to palatal implant positioning of the implants. Thus, immediate or early implant placement in the extraction socket has been suggested, because it would reduce the time period and the number of surgical intervention and yield higher patient satisfaction compared with delayed placed implants. However, placement of an implant immediately after tooth extraction may result in a gap between the occlusal portion of the implant and the surrounding alveolar bone crest. In this case report, an implant-supported restoration which is in harmony with the surrounding hard and soft tissue was created by the immediate implant placement with ridge augmentation in anterior region with high satisfaction from the patient.

Histological and radiographic evaluation of the muscle tissue of rats after implantation of bone morphogenic protein (rhBMP-2) in a scaffold of inorganic bone and after stimulation with low-power laser light.

Objective: The present study histologically and radiologically evaluates the muscle tissue of rats after implantation of bone morphogenic protein (rhBMP-2) in a natural inorganic bone mineral scaffold from a bull calf femur and irradiation with low-power light laser. Materials and Methods: The right and left hind limbs of 16 rats were shaved and an incision was made in the muscle on the face corresponding to the median portion of the tibia, into which rhBMP-2 in a scaffold of inorganic bone was implanted. Two groups of limbs were formed: control (G1) and laser irradiation (G2). G2 received diode laser light applied in the direction of the implant, at a dose of 8 J/cm2 for three minutes. On the 7th, 21st, 40th and 112th days after implantation, hind limbs of 4 animals were radiographed and their implants removed together with the surrounding tissue for study under the microscope. The histological results were graded as 0=absence, 1=slight presence, 2=representative and 3=very representative, with regard to the following events: formation of osteoid structure, acute inflammation, chronic inflammation, fibrin deposition, neovascularization, foreign-body granuloma and fibrosis. Results: There were no statistically significant differences in these events at each evaluation times, between the two groups (P>0.05; Mann-Whitney test). Nevertheless, it could be concluded that the natural inorganic bone matrix with rhBMP-2, from the femur of a bull calf, is a biocompatible combination. Conclusions: Under these conditions, the inductive capacity of rhBMP-2 for cell differentiation was inhibited. There was a slight acceleration in tissue healing in the group that received irradiation with low-power laser light.

Bone regeneration after demineralized bone matrix and castor oil (Ricinus communis) polyurethane implantation

Innocuous biocompatible materials have been searched to repair or reconstruct bone defects. Their goal is to restore the function of live or dead tissues. This study compared connective tissue and bone reaction when exposed to demineralized bovine bone matrix and a polyurethane resin derived from castor bean (Ricinus communis). Forty-five rats were assigned to 3 groups of 15 animals (control, bovine bone and polyurethane). A cylindrical defect was created on mandible base and filled with bovine bone matrix and the polyurethane. Control group received no treatment. Analyses were performed after 15, 45 and 60 days (5 animals each). Histological analysis revealed connective tissue tolerance to bovine bone with local inflammatory response similar to that of the control group. After 15 days, all groups demonstrated similar outcomes, with mild inflammatory reaction, probably due to the surgical procedure rather than to the material. In the polymer group, after 60 days, scarce multinucleated cells could still be observed. In general, all groups showed good stability and osteogenic connective tissue with blood vessels into the surgical area. The results suggest biocompatibility of both materials, seen by their integration into rat mandible. Moreover, the polyurethane seems to be an alternative in bone reconstruction and it is an inexhaustible source of biomaterial.

Diagnosis and treatment of extraction sockets in preparation for implant placement: report of three cases

This article addresses diagnostic parameters that should be assessed in the treatment of extraction sockets with dental implant placement by presenting three case reports that emphasize the relevance of the amount of remaining bone walls. Diagnosis was based on the analysis of clinical and radiographic parameters (e.g.: bone defect morphology, remaining bone volume, presence of infections on the receptor site). Case 1 presents a 5-wall defect in the maxillary right central incisor region with severe root resorption, which was treated with immediate implant placement. Cases 2 and 3 present, respectively, two- and three-wall bone defects that did not have indication for immediate implants. These cases were first submitted to a guided bone regeneration (GBR) procedure with bone graft biomaterial and membrane barriers, and the implants were installed in a second surgical procedure. The analysis of the preoperative periodontal condition of the adjacent teeth and bone defect morphology is extremely important because these factors determine the choice between immediate implant or GBR treatment followed by implant installation in a subsequent intervention.

Este artigo aborda os parâmetros clínicos que devem ser observados no diagnóstico e tratamento de alvéolos de extração por meio da instalação de implantes através da apresentação de casos clínicos, enfatizando a importância do número de paredes ósseas. O diagnóstico baseou-se na análise de parâmetros clínicos e radiográficos (p. ex: morfologia do defeito ósseo, volume ósseo remanescente, presença de infecção na área receptora). O Caso 1 apresenta um defeito de 5 paredes relacionado a um incisivo central superior direito que apresentava extensa reabsorção radicular e foi tratado através da técnica de implante imediato. Os Casos 2 e 3 consistem em defeitos de 2 e 3 paredes ósseas presentes, respectivamente, que inicialmente não devem ser tratados com implantes imediatos, e, por isso, foram submetidos inicialmente ao procedimento de regeneração óssea guiada, com uso de biomateriais para enxertia e barreiras de membrana, para receberem implantes numa segunda etapa cirúrgica. A avaliação prévia das condições periodontais dos dentes adjacentes e da morfologia do defeito ósseo são extremamente importantes, uma vez que determinam se determinada área será tratada pela técnica de ROG ou pela técnica de implantes imediatos.

Comparação histológica entre o osso desmineralizado e polímero de mamona sobre a regeneração óssea / Histological comparison of demineralized bone matrix and the Ricinus communis polymer on bone regeneration

O presente estudo tem por objetivo analisar histologicamente o efeito de uma matriz óssea desmineralizada de origem humana e de uma resina poliuretana derivada do óleo da mamona sobre o processo de reparação óssea. MATERIAIS E MÉTODOS: Foram utilizados 24 coelhos e em cada calvária foram preparadas duas cavidades cirúrgicas, sendo uma do lado direito e a outra do lado esquerdo da sutura parietal. Os animais foram divididos em dois grupos. No grupo I, a cavidade experimental (lado direito) foi preenchida com a matriz óssea desmineralizada de origem humana, enquanto no grupo II a cavidade experimental foi preenchida com a poliuretana derivada do óleo de mamona. As cavidades de controle foram preenchidas apenas com o sangue do animal. Os animais foram sacrificados nos períodos pós-operatórios de 4, 7 e 15 semanas. RESULTADOS: A análise histológica revelou que tanto o grupo controle quanto os grupos I e II apresentaram um aumento na neoformação óssea ao longo do tempo, sendo que esta reparação se deu mais rapidamente no grupo controle, mesmo mostrando diminuição importante na espessura. CONCLUSÕES: Ambos os materiais apresentaram-se biocompatíveis, sendo a poliuretana reabsorvida mais tardiamente e considerada de melhor resultado em relação ao outro material empregado.

Aim the aim of the present study is to make a histologic analysis the effects of a human demineralized bone matrix and a polyurethane resin derived from the Ricinus communis, on bone regeneration process. MATERIALS AND METHODS: For this, 24 rabbits were submitted to two surgical calvaria bone defects, one on the right and another on the left side of the parietal suture. The animals were divided in two groups. In group I the experimental defect (right side) was treated with a human demineralized bone matrix, while in group II the experimental cavity was treated with the polyurethane resin derived from Ricinus communis. The control defects were filled with the animalÆs own blood. The animals were slaughtered after subsequent periods of 04, 07 and 15 weeks. RESULTS: The histological analysis revealed that all groups (control and experiment), presented increased bone regeneration with time, but this repair was faster in the control group, even showing important decrease in defect thickness. CONCLUSION: Both materials proved to be biologically compatible, however polyurethane was more slowly resorbed presented considerable better results when compared with demineralized bone matrix.

Assessment of bovine biomaterials containing bone morphogenetic proteins bound to absorbable hydroxyapatite in rabbit segmental bone defects

PURPOSE: To evaluate the osteo-regenerative capacity of two proprietary bone grafting materials, using a segmental defect model in both radial diaphyses of rabbits. METHODS: The right defect was filled with pooled bone morphogenetic proteins (pBMPs) bound to absorbable ultrathin powdered hydroxyapatite (HA) mixed with inorganic and demineralized bone matrix and bone-derived collagen, derived from bovine bone (Group A). The left defect was filled with bovine demineralized bone matrix and pBMPs bound to absorbable ultrathin powdered HA (Group B). In both groups, an absorbable membrane of demineralized bovine cortical was used to retain the biomaterials in the bone defects, and to guide the tissue regeneration. The rabbits were euthanized 30, 90 and 150 days after surgery. Radiographic, tomographic and histologic evaluations were carried out on all specimens. RESULTS: At 30 days, the demineralized cortical bone cover was totally resorbed in both groups. HA was totally resorbed from Group A defects, whereas HA persisted in Group B defects. A prominent foreign body reaction was evident with both products, more pronounced in sections from Group B. At 90 days, the defects in Group B exhibited more new bone than Group A. However, at 150 days after surgery, neither treatment had stimulated complete repair of the defect. CONCLUSION: The partial bone healing of the segmental defect occurred with low or none performance of the biomaterials tested.

OBJETIVO: Avaliar a capacidade osteo-regenerativa de dois biomateriais utilizando um modelo de defeito segmentar efetuado nas diáfises do rádio de coelhos. MÉTODOS: O defeito direito foi preenchido com pool de proteínas morfogenéticas ósseas (pBMPs) e hidroxiapatita em pó ultrafina absorvível (HA) combinada com matriz óssea inorgânica desmineralizada e colágeno, derivados do osso bovino (Grupo A). O defeito esquerdo foi preenchido com matriz óssea desmineralizada bovina com pBMPs e hidroxiapatita em pó ultrafina absorvível (Grupo B). Em ambos os defeitos utilizou-se membrana reabsorvível de cortical bovina desmineralizada para reter os biomateriais no defeito ósseo e guiar a regeneração tecidual. Os coelhos foram submetidos à eutanásia aos 30, 90 e 150 dias após a cirurgia. Foram efetuados exames radiográficos, tomográficos e histológicos em todos os espécimes. RESULTADOS: Aos 30 dias de pós-cirúrgico, o osso cortical desmineralizado foi totalmente reabsorvido em ambos os grupos. A HA tinha reabsorvido nos defeitos do Grupo A, mas persistiu nos do Grupo B. Uma reação de corpo estranho foi evidente com ambos os produtos, porém mais pronunciada no Grupo B. Aos 90 dias os defeitos do grupo B tinham mais formação óssea que os do Grupo A. Entretanto, aos 150 dias após a cirurgia, nenhum tratamento havia promovido o completo reparo do defeito. CONCLUSÃO: Os biomateriais testados contribuíram pouco ou quase nada para a reconstituição do defeito segmentar.

Matriz óssea homóloga desmineralizada associada à medula óssea autógena fresca na artrodese vertebral dorsolateral lombar em coelhos / Homologous demineralized bone matrix associated to autologous bone marrow in a rabbit dorsolateral lumbar vertebral fusion

Foi avaliada a matriz óssea homóloga desmineralizada (MOD), associada à medula óssea autógena fresca (MO), na artrodese vertebral dorsolateral lombar em 24 coelhos (grupo 1), depositada sobre os processos transversos de L5-L6 previamente descorticados. Em outros nove coelhos (grupo-controle), foi realizada apenas a descorticação dos processos transversos e nas diferentes avaliações, foi observada insignificante reação periosteal local, sem união óssea. No grupo 1, às cinco semanas de pós-operatório, 87,5 por cento dos animais apresentaram união (artrodese) à palpação, seguido por 75 e 100 por cento às sete e nove semanas, respectivamente. Os índices de artrodese aos RX foram de 50 por cento, 62 por cento e 75 por cento, no mesmo período de avaliação. Histologicamente, às cinco semanas havia características de osteointegração da MOD com os processos transversos descorticados, de onde partiram os vasos sangüíneos que penetraram a matriz, e presença de ilhas de cartilagem no centro do enxerto. Nas semanas seguintes, a ossificação endocondral evoluiu e a MOD foi quase totalmente substituída por tecido ósseo trabecular, formando uma ponte de osso maduro entre e sobre os processos transversos adjacentes, propiciando ao segmento operado maior resistência mecânica em comparação aos segmentos adjacentes. A associação da MOD com a MO mostrou-se uma opção exeqüível e de baixo custo na artrodese vertebral dorsolateral lombar em coelhos.

Estudo do reparo ósseo com matriz óssea bovina desvitalizada e calcitonina em ratos / Study of bone repair with bovine bone devitalized matrix and calcitonin in rats

OBJETIVO: Estudar o efeito da calcitonina no reparo de defeito femoral preenchido com matriz óssea bovina desvitalizada. MÉTODOS: Quarenta e oito ratos Wistar, machos e adultos foram distribuídos aleatoriamente em dois grupo com 24 animais. Defeitos femorais bilaterais, com 2mm de diâmetro, foram produzidos na diáfise femoral e preenchidos com matriz óssea bovina desvitalizada. O grupo experimento recebeu 0,05mL de calcitonina, intramuscular, no pós-operatório imediato; o grupo controle recebeu 0,05mL de cloreto de sódio 0,9 por cento. Os animais foram sacrificados com 7, 14 e 21 dias e os fêmures submetidos a avaliação clínica, microscópica e histométrica. Foram utilizados os testes "t" de Student, Kruskal-Wallis e Mann-Whitney para a análise estatística dos resultados. RESULTADOS: Os resultados clínicos não mostraram diferença entre os grupos controle e experimento. Microscopicamente, os defeitos mostraram reação inflamatória e formação óssea ativa nas proximidades da matriz óssea bovina. A análise quantitativa mostrou maior formação óssea nas fases iniciais da osteogênese reparadora no grupo experimento (Z=4,82 e Z=4,43), sem diferenças em relação à reação inflamatória em ambos grupos. CONCLUSAO: O uso de calcitonina em defeitos preenchidos com matriz óssea bovina desvitalizada estimula a neoformação óssea com 7 e 14 dias de pós-operatório, mas não diminui a reação inflamatória.

Potencial osteoindutor da matriz óssea homóloga desmineralizada de coelho / Osteoinductivity of demineralized homologous bone graft in rabbits

O trabalho objetivou avaliar o potencial osteoindutor da matriz óssea homóloga (MOD) desmineralizada em ácido clorídrico. Em doze coelhos adultos, foi realizada uma incisäo sagital mediana na pele abdominal, de aproximadamente 5 cm, e a divulsäo do tecido subcutâneo, permitindo que três fragmentos de MOD, com 1 cm de comprimento, fossem implantados em "bolsas" confeccionadas cirurgicamente no músculo reto abdominal. A MOD foi preparada a partir de rádios de coelhos sacrificados por superdosagem anestésica e estocada em etanol 70 por cento, em temperatura ambiente, por, no mínimo, 15 dias, até o momento do uso. Para permitir análise prévia às enxertias, fragmentos de MOD preservados em etanol 70 por cento e fixados em formol a 10 por cento tamponado foram submetidos à rotina histológica. Quatro coelhos foram sacrificados a cada duas, quatro e seis semanas de pós-operatório. A rotina histológica foi realizada, também, em dois dos enxertos implantados em cada animal, em cortes longitudinais e transversais, corados por H&E e no fragmento restante a deposiçäo de osso novo foi evidenciada pela fluorescência da tetraciclina administrada aos animais. Os resultados da pesquisa permitiram concluir que o ácido clorídrico 0,6 N promoveu desmineralizaçäo eficaz e manteve o potencial indutor da matriz óssea; a imersäo da matriz óssea desmineralizada em etanol 70 por cento propiciou a esterilizaçäo dos enxertos homólogos e näo inibiu o potencial osteoindutor; e também que, a MOD, devido a fatores indutores inerentes que atuam sobre as células dos tecidos adjacentes, promoveu quimiotaxia, mitose e diferenciaçäo celular.

Matriz óssea homóloga desmineralizada na preparaçäo de falhas ósseas segmentares produzidas no rádio de coelhos / Repair of segmental defects produced in the radii rabbit by homologous demineralized bone matrix

No presente trabalho, foram avaliados os potenciais osteoindutor e osteocondutor da matriz óssea homóloga desmineralizada (MOD) em ácido clorídrico 0,6 N e conservada em etanol 70 por cento e sua utilizaçäo no preenchimento de falhas osteoperiosteais induzidas em rádio de coelhos. O rádio operado de todos os animais foi radiografado imediatamente após a cirurgia e na 3ª, 6ª, 9ª e 12ª semanas após. No grupo tratado, sete animais foram sacrificados a cada três semanas, sendo realizadas tomografia computadorizada e avaliaçöes macro e microscópica e da fluorescência óssea à tetraciclina. Os animais do grupo controle foram sacrificados na 12ª semana de pós-operatório. Foi constatado que a matriz acelerou o processo de formaçäo óssea, devido a fatores indutores inerentes que atuaram sobre as células dos tecidos adjacentes e que os canais vasculares, as lacunas e o canal medular vazios da MOD, serviram de arcabouço para a penetraçäo celular, determinando osteoconduçäo.

Emprego de matriz óssea orgânica bovina e hidroxiapatita no reparo de defeito induzido em crânio de ratos / Use of bovine organic bone matrix and hydroxyapatite in repair of skull induced defects of rats

OBJETIVO: Avaliar a resposta biológica e poder indutor da combinação matriz óssea orgânica bovina e hidroxiapatita. MÉTODOS: Foram produzidos defeitos ósseos em crânios de 20 ratos, Wistar, machos, com peso inicial médio entre 300 e 350 gramas, divididos eqüitativamente em dois grupos. No grupo I o defeito foi preenchido com uma associação de matriz óssea orgânica bovina e hidroxiapatita, ao passo que no grupo II considerado controle, nada foi empregado. Para o estudo histológico foram submetidos à eutanásia, cinco animais de cada grupo, aos 30 e 60 dias de pós-operatório. RESULTADOS: Com 30 dias de pós-operatório a reabsorção óssea foi maior no grupo II. A presença de cartilagem não diferiu entre os momentos de um mesmo grupo, porém houve diferença estatística entre os grupos I e II aos 30 dias, sendo maior no grupo I. Aos 60 dias de pós-operatório, o grupo I apresentava uma maior quantidade de osteoblastos e presença de osso comparativamente ao grupo II. CONCLUSÃO: A combinação matriz óssea orgânica bovina e hidroxiapatita não causa reações adversas e favorece a reparação óssea.

Effect of a persistent inflammatory process on experimental heterotopic ossification: the influence of macrophages

1. We investigated the effect of a persistent carrageenin- or nystatin-induced inflammatory reaction on heterotopic ossification produced by the subcutaneous implant of a demineralized bone matrix in female Swiss mice (25 to 35 g). 2. Subcutaneous carrageenin injection (0.3 ml of a 2 per cent solution in saline) into mice induced an inflammatory reaction characterized by a mature granuloma predominantly of macrophages containing particles of the irritant in their cytoplasm and which remained unchanged until the end of the experiment (40th day). 3. Subcutaneous nystatin inoculation (30,000 IU in 0.3 ml saline) induced an inflammatory reaction consisting initially of macrophages (4th day) but later turning into an epithelioid granuloma (7th day) consisting predominantly of epithelioid cells and which was present up to the 21st day when it was gradually replaced by adipocytes up to the 30th day. 4. An intramuscular implant of demineralized bone matrix (DBM, approximately 10 mg) induced the formation of cartilage and bone tissue and of hemopoietic bone marrow (heterotopic ossification) in 100 per cent of the control animals (N = 5). An intramuscular DBM implant in animals that received carrageenin (N = 19) or nystatin (N = 21) induced heterotopic ossification in 100 and 57 per cent (P < 0.01) of the animals, respectively. 5. The response to a dorsal subcutaneous DBM implant was essentially negative in control animals (N = 5), whereas implants performed near the site injected with carrageenin (N = 28) or nystatin (N = 31) produced a response in 71 (P < 0.01) and 36 per cent (P < 0.01) of the animals, respectively. A DBM implant into the contralateral (control) dorsal subcutaneous tissue of the same animals that received carrageenin (N = 25) or nystatin (N = 29) resulted in heterotopic ossification in 64 (P < 0.01) and 7 per cent of the animals, respectively. 6. The results suggest that the macrophages present in the mature granuloma induced by carrageenin somehow favored the development of metaplastic plates after subcutaneous DBM implant and that this effect may be systemic since the same response was observed in contralateral subcutaneous tissue