Tomographic, Histological, and Immunohistochemical Evidences on the Use of N-Butyl-2-Cyanoacrilate for Onlay Graft Fixation in Rabbits

Background: The bone tissue responses to Cyanoacrylate have been described in the literature, but none used N-butyl-2-cyanoacrilate (NB-Cn) for bone graft fixation. Purpose: The aims of the study were: (a) to analyze the bone grafts volume maintenance fixed either with NB-Cn or titanium screw; (b) to assess the incorporation of onlay grafts on perforated recipient bed; and (c) the differences of expression level of tartrate-resistant acid phosphatase (TRAP) protein involved in bone resorption. Materials and Methods: Eighteen New Zealand White rabbits were submitted to calvaria onlay grafting on both sides of the mandible. On one side, the graft was fixed with NB-Cn, while on the other hand the bone graft was secured with an osteosynthesis screw. The computed tomography (CT) was performed just after surgery and at animals sacrifice, after 1 (n=9) and 6weeks (n=9), in order to estimate the bone grafts volume along the experiments. Histological sections of the grafted areas were prepared to evaluate the healing of bone grafts and to assess the expression of TRAP protein. Results: The CT scan showed better volume maintenance of bone grafts fixed with NB-Cn (p≤0.05) compared with those fixed with screws, in both experimental times (analysis of variance). The immunohistochemical evaluation showed that the TRAP expression in a 6-week period was significantly higher compared with the 1-week period, without showing significant difference between the groups (Wilcoxon and Mann-Whitney). Histological analysis revealed that the NB-Cn caused periosteum damage, but provided bone graft stabilization and incorporation similar to the control group. Conclusion: The perforation provided by screw insertion into the graft during fixation may have triggered early revascularization and remodeling to render increased volume loss compared with the experimental group. These results indicate that the NB-Cn possesses equivalent properties to titanium screw to be used as bone fixation material in osteosynthesis. © 2010 Wiley Periodicals, Inc.

Block Bone Graft Fixation ( Onlay): A Modification of the Surgical Technique

Several reconstructive methods of the alveolar ridge have been reported to make possible future rehabilitations with implants. Many of these methods come from studies done in animals, mainly rats. With this clinical practice based on scientific evidence, any experimental procedure that can be undertaken in real life is fundamental. Thus, any research that emulates as closely as possible those techniques used in humans are important. This study describes the modification of the technique for block bone graft fixation (onlay) in rats using the lag screw-type technique, normally used in clinical procedures for grafts in humans. The conclusion was that the execution of the described procedures minimizes interference of blood flow in the area because of the maintenance of the muscle insertion in the buckle aspect of the most anterior region of the mandible, providing better stability to the graft and better contact interface of the graft and receptor bed.

Utilization of Ethyl Cyanoacrylate and 2-Octyl Cyanoacrylate Adhesives for Autogenous Bone Graft Fixation: Histomorphometric Study in Rats

The present study analyzes the repair process of autogenous bone graft in a block fixed with ethyl cyanoacrylate and 2-octyl cyanoacrylate adhesives in rat calvaria. Forty-eight rats, divided into 3 groups, received round osteotomies at the right parietal bone for the attainment of autogenous bone graft fragment, which was fixed at the opposite side to the donor site with ethyl cyanoacrylate (ethyl group) and 2-octyl cyanoacrylate (octyl group) adhesives. In the control group, bone fragment was only juxtaposed at the parietal bone surface without any fixation material. The animals were euthanized after 10 and 60 postoperative days. The calvariae were processed in a laboratory for the attainment of slides stained through the hematoxylin and eosin technique for histological and histometric analysis. The qualitative analysis showed a discrete inflammatory infiltrate in the control group and moderate inflammatory infiltrate in the ethyl and octyl groups at the 10-day period, which remained at the 60-day period, mainly in the octyl group. The bone fragment remained bonded to the recipient site through the adhesive, but graft incorporation was not observed in any of the specimens. Resorption was higher in the octyl group followed by the ethyl and control groups, both at the 10-and 60-day periods, but with no statistical significance (P < .05). Although promoting graft fixation and its maintenance at the recipient site, both studied adhesives did not allow the graft incorporation, producing a localized and discrete inflammatory reaction, which persisted at 60 days, being more intense in the octyl cyanoacrylate group.

Reconstrução do L.C.P. do joelho: técnica de fixação no leito tibial (INLAY). Avaliação objetiva e subjetiva de 30 casos

A reconstrução cirúrgica do ligamento cruzado posterior (L.C.P.) do joelho ainda permanece como um grande desafio terapêutico. Neste trabalho avaliamos 30 pacientes submetidos à reconstrução cirúrgica do L.C.P. com a técnica de fixação do enxerto tendíneo no leito tibial por abordagem direta (INLAY). 28 pacientes eram do sexo masculino e 2 do feminino, com idade média de 31,10 anos. O tempo médio de lesão foi de 34,24 meses em 67% dos casos a lesão foi secundária a acidente motociclístico. As lesões condrais e do ligamento cruzado anterior (L.C.A.) do joelho estavam presentes em 67% e 33% dos casos, respectivamente. Os pacientes foram avaliados objetivamente (teste de gaveta posterior) e subjetivamente (Escala de Lysholm). O seguimento pós-operatório médio foi de 21,7 meses. Cerca de 66% dos casos foram classificados como bom e excelente na avaliação subjetiva e objetiva. A análise estatística apresentou comportamento semelhante para as duas avaliações.Os resultados clínicos pós-operatórios obtidos neste trabalho têm nos encorajado a seguir com esta técnica cirúrgica.

Comparação entre parafuso de interferência e transcondilar na reconstrução do LCA

OBJETIVO: A fixação femoral na reconstrução do Ligamento Cruzado Anterior (LCA) com enxerto de tendões flexores pode variar muito de acordo com a disposição dos materiais e a experiência do cirurgião. Porém os trabalhos que comparam os sistemas de fixação são efetuados na maioria das vezes em cadáveres ou em animais, sem avaliar os resultados clínicos, prejudicando sua comparação com pacientes humanos vivos. Neste artigo avaliamos o resultado clínico de dois métodos de fixação do enxerto ao fêmur (parafuso de interferência de titânio e parafuso transcondilar de titânio) para saber se, do ponto de vista clinico e subjetivo, há diferença entre estes métodos. MÉTODOS: Foram selecionados 40 pacientes com lesão do LCA sendo que 20 pacientes tiveram seu enxerto fixado ao fêmur com parafuso de interferência e 20 com parafuso transcondilar. Todos foram reavaliados com no mínimo dois anos de pós operatório para medição da gaveta anterior, Pivot Shift e teste de Lachman, além da obtenção dos questionário de Lysholm e IKDC (International Knee Documentation Committee). RESULTADOS: Os resultados não foram estatisticamente diferentes para os critérios avaliados. CONCLUSÃO: As duas formas de fixação são eficientes para esta técnica dentro dos parâmetros estabelecidos. Nível de Evidência II, Estudo Prospectivo Comparativo.

Miniplate fixation for the repair of segmental mandibular defects filled with autogenous bone in cats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fixation of autogenous bone grafts with ethyl-cyanoacrylate glue or titanium screws in the calvaria of rabbits

This study compared the fixation of autogenous onlay bone grafts with cyanoacrylate glue (Super Bonder) and with titanium screws. Twenty rabbits underwent bilateral parietal ostectomies. Bone segments were fixed anteriorly to the resulting bone defect. In group I, the grafts were fixed with 4 min long, 1.5 mm diameter screws; in group II, adhesive was used. The animals were killed after 5, 15, 30, 60 and 120 days. Histomorphometric analysis was used to quantify the maintenance of the graft area. Discrete areas of inflammatory reaction were seen in both groups after 5 days and for group II after 15 days. After 30 days, new bone formation was seen at the interface of the grafts. After 120 days, the graft was incorporated into the host bed in group I and partially incorporated in group II. There was a significant statistical difference regarding the mean graft areas between 15 and 120 days (p < 0.001) and between fixation methods (p < 0.002). Fixation with adhesive promoted a significantly greater area of bone graft than screw fixation, independent of time period. The adhesive was biocompatible, presented similar stability to the screw and maintained the bone area, although there was a delay in graft incorporation.

Immunohistochemical, tomographic, and histological study on onlay bone graft remodeling. Part III: allografts

Objective In the last decades aroused the interest for bone tissue bank as an alternative to autogenous grafting, avoiding donor sites morbidity, surgical time, and costs reduction. The purpose of the study was to compare allografts (ALg) with autografts (AUg) using histology, immunochemistry, and tomographic analysis. Material and methods Fifty-six New Zealand White rabbits were submitted to surgical procedures. Twenty animals were donors and 36 were actually submitted to onlay grafting with ALg (experimental group) and AUg (control group) randomly placed bilaterally in the mandible. Six animals of each group were sacrificed at 3, 5, 7, 10, 20, and 60 postoperative days. Immunolabeling was accomplished with osteoprotegerin (OPG); receptor activator of nuclear factor-k ligand (RANKL); alkaline phosphatase (ALP); osteopontin (OPN); vascular endothelial growth factor (VEGF); tartrate-resistant acid phosphatase (TRAP); collagen type I (COL I); and osteocalcin (OC). Density and volume of the grafts was evaluated on tomography obtained at the surgery and sacrifice. Results The ALg and AUg exhibited similar patterns of density and volume throughout the experiments. The intra-group data showed statistical differences at days 7 and 60 in comparison with other time points (P = 0.001), in both groups. A slight graft expansion from fixation until day 20 (P = 0.532) was observed in the AUg group and then resorbed significantly at the day 60 (P = 0.015). ALg volume remained stable until day 7 and decreased at day 10 (P = 0.045). The light microscopy analysis showed more efficient incorporation of AUg onto the recipient bed if compared with the ALg group. The immunohistochemical labeling picked: at days 10 and 20 with OPG in the AUg group and at day 7 with TRAP in the ALg group (P = 0.001 and P = 0.002, respectively). Conclusions ALg and AUg were not differing in patterns of volume and density during entire experiment. Histological data exhibit more efficient AUg incorporation into recipient bed compared with the ALg group. Immunohistochemistry outcomes demonstrated similar pattern for both ALg and AUg groups, except for an increasing resorption activity in the ALg group mediated by TRAP and in the AUg group by higher OPG labeling. However, this latter observation does not seem to influence clinical outcomes.

Reconstruction of maxillary ridge atrophy caused by sequela of dentoalveolar trauma with autogenous bone graft harvested from mentum

Dentoalveolar traumatisms, particularly those that affect the anterior teeth, interfere adversely in the patient s life.Among them, tooth avulsion is pointed out because it is characterized as a complex injury that affects multiple tissues, andbecause there is no effective treatment available for its resolution with a stable long-term outcome.Aim/Hypothesis: The aim of the present study was to relate a clinical case of complete reconstruction of atrophy of the alveolarbone corresponding to tooth 11, lost by tooth resorption 10 years after the tooth reimplantation procedure.Material and methods: Reconstruction was performed with autogenous bone harvested from the mentum donor site. Surgicalaccess began in the receptor area with a Newman mucoperiosteal incision using a scalpel blade 15 mounted in a scalpel handlefor detachment and exposure of the receptor site. Extensive bone resorption was observed in the vestibular-palatine direction,proved by the thinness of the receptor bed. Decorticalization of the vestibular bone plate was performed. After preparing thereceptor bed, and incision was made in the mucosa in the depth of the anterior vestibular fornix, then a perpendicular muscleperiostealincision to detach and exposure the donor area. The bone graft necessary for reconstruction of the donor area wasdelimited, followed by monocortical osteotomy and the monocortical graft was removed. The next stage was to perform shapingfor passive graft accommodation and fixation by means of two bicortical screws. After fixation of the graft the sharp angles wererounded off in order to avoid possible exposure and/or fenestrations of the reconstructed area, then the receptor and donor areawere sutured. After the 6-month period to allow incorporation of the autogenous graft, an osseointegrated dental implant wasinserted. At the end of the 6-month period of waiting for osseointegration to occur, the process of fabricating the screw-retainedmetal ceramic

A comparison of two agar gel immunodiffusion methods and a complement fixation test for serologic diagnosis of Brucella ovis infection in experimentally infected rams

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Genetic characterization and nitrogen fixation capacity of Rhizobium strains on common bean

O objetivo deste trabalho foi a caracterização genética de quatro novas estirpes de Rhizobium e a avaliação de sua capacidade de fixação de N2 e nodulação, comparadas a estirpes comerciais e à população nativa de rizóbios de um Latossolo Vermelho. Dois experimentos foram conduzidos em blocos ao acaso, em casa de vegetação. No primeiro experimento, conduzido em tubetes com vermiculita, avaliaram-se a nodulação e a capacidade de fixação das novas estirpes, em comparação com as estirpes comerciais CIAT-899 e PRF-81 e com a população nativa do solo. Das colônias puras isoladas, extraiu-se o DNA genômico e realizou-se o seqüenciamento do espaço intergênico, para a caracterização genética das estirpes e da população nativa de rizóbios. O segundo experimento foi realizado em vasos com solo, para determinação da produtividade e da nodulação do feijoeiro, cultivar Pérola, com o uso das estirpes isoladamente ou em mistura com a PRF-81. A população nativa do solo foi identificada como Rhizobium sp. e se mostrou ineficiente na fixação de nitrogênio. Foram encontradas três espécies de Rhizobium entre as quatro novas estirpes. As estirpes LBMP-4BR e LBMP-12BR estão entre as que têm maior capacidade de nodulação e fixação de N2, e apresentam respostas diferenciadas quando misturadas à PRF-81.

Study of the carbon dioxide chemical fixation-activation by guanidines

Fixation of CO2 is one of the most important priorities of the scientific community dedicated to reduce global warming. In this work, we propose new methods for the fixation of CO2 using the guanidine bases tetramethylguanidine (TMG) and 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-a]-pyrimidine (TBD). In order to understand the reactions occurring during the CO2 fixation and release processes, we employed several experimental methods, including solution and solid-state NMR, FTIR, and coupled TGA-FTIR. Quantum mechanical NMR calculations were also carried out. Based on the results obtained, we concluded that CO2 fixation with both TMG and TBD guanidines is a kinetically reversible process, and the corresponding fixation products have proved to be useful as transcarboxylating compounds. Afterward, CO2 thermal releasing from this fixation product with TBD was found to be an interesting process for CO2 capture and isolation purposes. (C) 2008 Elsevier Ltd. All rights reserved.

Selectivity of the cyclic carbonate formation by fixation of carbon dioxide into epoxides catalyzed by Lewis bases

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Graft versus host disease in a rat small bowel transplant model after T-cell depleted donor specific bone marrow infusion

Baixas doses de irradiação associadas à infusão de células da medula óssea não previnem a ocorrência da reação do enxerto versus hospedeiro após o transplante intestinal. OBJETIVO: Neste estudo foi avaliado a potencial vantagem em estender o regime imunossupressor associado a infusão de células de medula óssea do doador depletadas de células T na prevenção da reação do enxerto versus hospedeiro após o transplante intestinal. MÉTODOS: Transplante heterotópico de intestino delgado foi realizado em ratos Lewis como receptores e da como doadores, distribuídos em cinco grupos de acordo com a duração da imunossupressão, irradiação e do uso de medula óssea normal ou depletada: G1 (n=6), sem irradiação e G2 (n=9), G3 (n=4), G4 (n=5) e G5 (n=6) foram irradiados com 250 rd. Grupos1, 2, 4 e G3 e 5 foram infundidos com 100 x 10(6) células da medula normal e depletada respectivamente. Animais no G1,2,3 foram imunossuprimidos com 1mg/kg/FK506/ IM por cinco dias e G4 e cinco por 15 dias. Anticorpos monoclonais contra células CD3 e colunas magnéticas foram utilizadas para a depleção da medula óssea. Os animais foram examinados para a presença de rejeição, reação do enxerto versus hospedeiro, chimerismo e biópsias intestinais e da pele. RESULTADOS: Rejeição mínima foi observada em todos os grupos; entretanto, a reação do enxerto versus hospedeiro somente nos animais irradiados. Extensão da imunossupressão alterou a gravidade da reação nos animais dos G4 e 5. Rejeição foi a causa mortis no G1 e a reação do enxerto versus hospedeiro nos Grupos 2,3,4 e 5, não controlada com a infusão de medula óssea depletada. O chimerismo total e de células T do doador foi estatisticamente maior nos grupos irradiados em comparação ao G1. CONCLUSÃO: A extensão do regime de imunossupressão associado a baixas doses de irradiação diminui a gravidade da reação do enxerto versus hospedeiro, não abolida pelo uso de medula óssea depletada.

Muscle reinnervation in one or two stages?: experimental study in rats with end-to-side nerve graft

OBJETIVO: Comparar a reinervação muscular com enxerto de nervo em um e dois tempos operatórios, utilizando a neurorrafia término-lateral (NTL) sem lesão do nervo doador. MÉTODOS: Vinte ratos foram distribuídos em quatro grupos. O grupo 1 (G1), um estágio, recebeu o enxerto que foi suturado ao nervo tibial (NT), por meio de NTL, e seu coto livre foi suturado por NTL ao coto distal do nervo peroneal (NP), seccionado a um centímetro do NT, na mesma cirurgia. O grupo 2 (G2), dois estágios, recebeu o enxerto de nervo na primeira cirurgia, como já descrito. Dois meses depois, na segunda cirurgia, o NP foi seccionado e seu coto distal ligado ao coto distal do enxerto como em G1. O grupo controle de normalidade (Gn) recebeu o enxerto da mesma forma, apenas. E o grupo controle de denervação (Gd), além de receber o enxerto, teve o NP seccionado e seus cotos sepultados na musculatura adjacente, com a finalidade de denervar o músculo tibial cranial (MTC), alvo deste estudo. Os parâmetros utilizados para avaliar a reinervação do MTC foram massa muscular, diâmetro mínimo da fibra muscular e área. RESULTADOS: O grupo G2 apresentou superioridade (p<0,0001) em relação ao G1 na massa do MTC, no diâmetro mínimo e na área das fibras musculares. Na comparação entre os quatro grupos, estes mesmos parâmetros tiveram sua expressão máxima em Gn e mínima em Gd, como era esperado. CONCLUSÃO: A reinervação muscular em dois estágios apresenta melhor resultado quando comparada à técnica em um tempo.