Integration of heterologous bone matrix associated with polymethylmethacrylate in induced tibial bone defects. An experimental study in rabbits

ABSTRACT Purpose To analyze and compare the reactions at the interface between the composite, composed of fragmented heterologous mineralized bone matrix (MOMHF) and polymethylmethacrylate (PMMA), and the rabbit's tibias, through macroscopic evaluation and scanning electron microscopy (SEM) in different periods. Methods In this study, 12 New Zealand adult rabbits were used (E1: n = 3, E2: n = 3, E3: n = 3 and E4: n = 3). They had the right tibial defects filled with composite and were evaluated immediately after surgery and at 30, 60, 90, and 120 days. Results The composites were incorporated and integrated into the recipient beds in 100% of the cases, defined by the MOMHF osseointegration and the PMMA fibrointegration, with no sign of infection, migration, or rejection. Conclusions The behavior of the composites in the recipient beds demonstrates that these biomaterials have the potential to be used in bone defect repairs, offering, thus, better quality of life to the orthopedic patient.

Região metafisária da tíbia de coelhos: uma opção para estudos dos substitutos ósseos / Metaphyseal region of the tibia of rabbits: an option for bone substitutes studies

Apresenta-se um modelo experimental de ostectomia de tíbia em coelhos para estudo de biomateriais em processos de reparação óssea. Foi realizada falha segmentar de 6mm de diâmetro na região metafisária medial proximal de tíbia de 72 coelhos as quais foram preenchidas com substituto ósseo e avaliadas clinicamente, por exame radiográfico e por meio de tomografia computadorizada feixe cônico, em diferentes tempos. Conclui-se que a metáfise medial proximal de tíbia de coelhos é adequada como modelo para estudos que avaliem o comportamento de enxertos e/ou biomateriais em falhas ósseas.

Is presented an experimental model of tibial ostectomy in rabbits to study biomaterials during bone repair process. Segmental failure of 6 mm diameter was performed in the medial proximal tibial metaphyseal region of 72 rabbits, which were filled with bone substitute and evaluated by clinical exam, X-ray, and cone beam computed tomography at different times. It is concluded that the medial proximal tibial metaphysis region of rabbits is suitable as a model for studies that evaluate the behavior of grafts and/or biomaterials on bone defects.

Região metafisária da tíbia de coelhos: uma opção para estudos dos substitutos ósseos / Metaphyseal region of the tibia of rabbits: an option for bone substitutes studies

Apresenta-se um modelo experimental de ostectomia de tíbia em coelhos para estudo de biomateriais em processos de reparação óssea. Foi realizada falha segmentar de 6mm de diâmetro na região metafisária medial proximal de tíbia de 72 coelhos as quais foram preenchidas com substituto ósseo e avaliadas clinicamente, por exame radiográfico e por meio de tomografia computadorizada feixe cônico, em diferentes tempos. Conclui-se que a metáfise medial proximal de tíbia de coelhos é adequada como modelo para estudos que avaliem o comportamento de enxertos e/ou biomateriais em falhas ósseas.

Is presented an experimental model of tibial ostectomy in rabbits to study biomaterials during bone repair process. Segmental failure of 6 mm diameter was performed in the medial proximal tibial metaphyseal region of 72 rabbits, which were filled with bone substitute and evaluated by clinical exam, X-ray, and cone beam computed tomography at different times. It is concluded that the medial proximal tibial metaphysis region of rabbits is suitable as a model for studies that evaluate the behavior of grafts and/or biomaterials on bone defects.

Intra-articular concentration of gentamicin administered by intravenous regional limb perfusion in healthy horses / Concentração intra-articular de gentamicina administrada por perfusão regional intravenosa em membros de equinos sadios

ABSTRACT: Intravenous regional limb perfusion (IRLP) is an efficient method to treat horses with synovial infections. However, information on the dose, volume, and intervals of administration is lacking. The aim of this study was to evaluate the intra-articular concentration of gentamicin administered by IRLP in horses over 24 h post-administration and evaluate the influence of the total perfused volume. Twenty horses were assigned to two treatment groups. Gentamicin 60 group (G60, n=10) and Gentamicin 250 group (G250, n=10) received IRLP of 6.6 mg/kg of gentamicin diluted in Ringer's lactate for a total volume of 60 ml (G60) and 250 ml (G250), respectively. Synovial fluid harvests were performed in the metacarpophalangeal joint before (0) and 4, 8, 12, 16, 20, and 24 hours after IRLP. Agar diffusion was the assay for measuring gentamicin concentrations. Four hours after IRLP, the intra-articular concentration of gentamicin was 47.58±49.21 μg/ml in G60 and 10.92±6.11 μg/ml in G250. During the 24 h of harvest, the intra-articular concentration of gentamicin remained above the minimum inhibitory concentration for horses (MIC; 2 μg/ml) in both experimental groups. In G60, the concentration of gentamicin in synovial fluid reached the maximal concentration and remained 8 to 10-fold higher than the MIC (Cmax: 16 to 20 μg/ml) during 12 h after IRLP. These findings support that IRLP with 6.6 mg/kg of gentamicin diluted to a volume of 60 ml promotes intra-articular concentrations higher than 250 ml, remaining above the MIC for 24 h and at the maximum concentration for 12 h.

RESUMO: A perfusão regional intravenosa (PRI) em membros de equinos é um método comprovadamente eficiente para o tratamento de infecções sinoviais, porém ainda existem lacunas quanto à dose, volume e intervalos de administração a serem utilizados. Neste estudo, objetivou-se avaliar a concentração intra-articular da gentamicina administrada por PRI em equinos, ao longo de 24 horas pós-administração, bem como avaliar a influência do volume total perfundido. Vinte equinos foram distribuídos em dois grupos experimentais, Grupo Gentamicina 60 (G60) e Grupo Gentamicina 250 (G250), representados por 10 equinos em cada grupo que receberam, por PRI, 6,6 mg/kg de gentamicina diluída em Ringer lactato para um total de 60 ml (G60) ou 250 ml (G250). As colheitas de líquido sinovial foram realizadas, na articulação metacarpo-falangeana, antes do início do experimento e após 4, 8, 12, 16, 20, e 24 horas. Difusão em ágar foi o método para doseamento das concentrações de gentamicina. Quatro horas após a PRI, a concentração intra-articular da gentamicina era de 47,58±49,21 µg/ml no G60 e 10,92±6,11 µg/ml no G250. Durante as 24 horas de colheita, a concentração intra-articular de gentamicina permaneceu acima da concentração inibitória mínima (CIM) (2 μg/ml) em ambos os grupos experimentais. No G60, as concentrações de gentamicina no líquido sinovial atingiram a concentração máxima e mantiveram-se 8 a 10 vezes mais elevadas que a CIM (Cmáx: 16 a 20 μg/ml) durante 12 horas após a PRI. Conclui-se que a PRI em membros, com 6,6 mg/kg de gentamicina diluída para o volume de 60 ml, promove concentrações intra-articulares mais elevadas que o volume de 250 ml, permanecendo acima da CIM durante 24 horas e em concentração máxima durante 12 horas.

Evaluation of polyamide surgical mesh as an abdominal ventral implant in rabbits

Abstract Purpose: To evaluate abdominal ventral wound healing by using a specific biomaterial, a handmade polyamide surgical mesh. Methods: A surgical incisional defect was made in ten rabbits to simulate a hernia in the ventral abdominal musculature. A polyamide surgical mesh was used in hernioplasty. They were monitored for surgical wound healing, and macroscopically and histologically evaluated at the end of the experiment. The polyamide surgical mesh did not cause foreign body reaction, pain, edema, or infection in the surgical site. The manure production was not affected by intestinal tissue adherences to the mesh, consistent with the ultrasonography result where adherences were not observed and organized scarring tissue formed in the incisional defect. The polyamide mesh was fixed over the abdominal wall, and its external and internal sides were surrounded by a vascularized connective tissue. Results: None of the experimental animals developed adherences from internal organs to the polyamide mesh, except two rabbits where the omentum formed adherence to the internal scarring tissue without present herniation or compromise of the rabbit's health. Conclusion: Polyamide surgical mesh for hernioplasty presents, in rabbits, excellent biocompatibility, with minimal body adverse reactions and low cost.

Didatic approach of ultrasonographic examination for evaluation of the carpal joint in horses / Abordagem didática do exame ultrassonográfico da articulação do carpo em equinos

ABSTRACT: The carpus is a complex articulation, which is often involved in injuries in equine athletes. The objective of this study was to suggest a didactic approach for ultrasonography training for the examination of the carpal joint in horses. Ultrasonograhy examination was performed in a healthy 14-year-old horse. The images were compared with those of a dissected anatomic specimen of the carpal region of a senior horse and with those reported in the literature. Identifiable structures were as follows: (dorsal) tendon of the muscle extensor carpi radialis, tendon of the muscle extensor carpi obliquus, tendon of the muscle commom digital extensor, dorsal synovial outpouchings, joint capsule and fat cushion, (lateral) tendon of the muscle lateral digital extensor, lateral collateral ligament (deep and superficial components), (medial) medial collateral ligament (deep and superficial components) and (palmar) palmar intercarpal ligament, carpal sheath, carpal tunnel, superficial digital flexor tendon, proximal origin of the deep digital flexor tendon, and palmar carpal ligament. Prior knowledge of the anatomy in combination with the study of anatomical specimens is essential for recognizing musculoskeletal structures during ultrasound examination, contributing to the training and learning processes.

RESUMO: O carpo é uma articulação complexa que, com frequência, é sede das doenças que acometem cavalos atletas. O objetivo deste estudo foi sugerir uma abordagem didática para o treinamento de exame ultrassonográfico do carpo em equinos. Foi realizado o exame ultrassonográfico de um equino hígido de catorze anos, as imagens foram comparadas com uma peça anatômica dissecada da mesma região de um cadáver de equino sênior e com dados da literatura. As estruturas passíveis de identificação foram: (dorsal) tendão do músculo extensor carporradial, tendão do músculo extensor oblíquo do carpo, tendão do músculo extensor digital comum, recessos sinoviais dorsais, cápsula articular, coxim de gordura, (lateral) tendão do músculo extensor digital lateral, ligamento colateral lateral (componente profundo e superficial), (medial) ligamento colateral medial (componente profundo e superficial), (palmar) ligamento intercarpiano palmar, bainha carpal, canal carpal, tendão do músculo flexor digital superficial, origem proximal do tendão do músculo flexor digital profundo e o ligamento carpal palmar. O conhecimento prévio da anatomia associada ao estudo em peças anatômicas são elementos essenciais para o reconhecimento das estruturas musculoesqueléticas ao ultrassom, contribuindo para o treinamento e processo de aprendizagem.

Tomographic imaging of fragmented cortical bone heteroimplant and methylmethacrylate in segmental bone defect of rabbit tibia

PURPOSE: To evaluate the performance of composites consisting of fragmented cortical bone heteroimplant in association with methylmethacrylate preserved in 98% glycerin, in segmental bone defect of rabbit tibia medial metaphysis. METHODS: In this study were used twelve adult New Zealand rabbits, divided into three groups of four animals each: G30 (30 days), G60 (60 days) and G90 (90 days). The bone defects previously created in the tibia were filled with composites and both were evaluated by cone-beam computed tomography, immediately after surgery and after 30, 60, and 90 days. RESULTS: The composites fulfilled and remained in the sites of bone defects in all cases and were not registered signals of infection, migration or rejection. CONCLUSIONS: The implanted composites promoted the bone defects repair without signals of infection and/or rejection. The composites are one more option for bone defects repair. .

Tomographic imaging of fragmented cortical bone heteroimplant and methylmethacrylate in segmental bone defect of rabbit tibia

PURPOSE: To evaluate the performance of composites consisting of fragmented cortical bone heteroimplant in association with methylmethacrylate preserved in 98% glycerin, in segmental bone defect of rabbit tibia medial metaphysis. METHODS: In this study were used twelve adult New Zealand rabbits, divided into three groups of four animals each: G30 (30 days), G60 (60 days) and G90 (90 days). The bone defects previously created in the tibia were filled with composites and both were evaluated by cone-beam computed tomography, immediately after surgery and after 30, 60, and 90 days. RESULTS: The composites fulfilled and remained in the sites of bone defects in all cases and were not registered signals of infection, migration or rejection. CONCLUSIONS: The implanted composites promoted the bone defects repair without signals of infection and/or rejection. The composites are one more option for bone defects repair.

Expression of matrix metalloproteases-2 and -9 in horse hoof laminae after intestinal obstruction, with or without Hydrocortisone treatment / Expressão de metaloproteinases 2 e 9 no tecido laminar do casco de equinos após obstrução intestinal e tratamento com hidrocortisona

Twenty horses were used in the experiment, for composed control group, (Cg) instrumented group, (Ig;without intestinal obstruction), treated group (Tg;submitted to intestinal obstruction and hydrocortisone treatment) and non-treated group (Ntg;submitted to intestinal obstruction without treatment). Immunohistochemistry and zymography techniques were used for researches on MMPs 2 and 9 in horse hoof laminae. There was an increase in the expression of MMP-2 in animals of Tg and Ntg. MMP-9 increased on animals from groups Ntg and Ig, however there was no rise of this MMP on the Tg when compared to the other groups in the immunohistochemistry analysis. Based on the results, it was observed that the intestinal injury caused by enterotomy and intestinal obstruction raise the quantities of MMPs in the hoof laminae.

Vinte cavalos foram usados no experimento: para compor o grupo controle (Cg), grupo instrumentado, Ig (sem obstrução intestinal), grupo tratado, Tg (submetidos à obstrução intestinal e tratamento com hidrocortisona) e grupo não tratado, Ntg (submetidos à obstrução intestinal, sem tratamento. Técnicas de zimografia e imunoistoquímica foram utilizadas para pesquisa de MMP-2 e MMP-9 no tecido laminar do casco dos equinos. Houve um aumento na expressão de MMP-2 nos animais dos grupos Tg e Ntg. A MMP-9 aumentou nos animais dos grupos Ig e Ntg. Houve aumento desta MMP no Tg quando comparado aos demais grupos na análise por zimografia. Observou-se que a injúria intestinal, causada pela enterotomia e obstrução intestinal, eleva a quantidade de MMPs no tecido laminar do casco.

Anestesia por isofluorano em eqüinos submetidos à infusão contínua de medetomidina ou xilazina / Isoflurane anesthesia in horses during medetomidine or xilazine continuous infusion

Avaliaram-se oito eqüinos sob anestesia geral inalatória com isofluorano (1CAM) e infusão contínua de xilazina (0,35mg kg-1h-1) ou medetomidina (3,5µg kg-1h-1), em relação à freqüência cardíaca, ritmo cardíaco, freqüência respiratória, pressão arterial, hemogasometria arterial e temperatura, nos tempos T0 (imediatamente antes do início da infusão contínua) e T10 ao T60 (intervalos de 10 minutos, após início da infusão contínua). Houve redução da freqüência cardíaca e da temperatura e elevação da pressão arterial média. A paCO2 (no GM) elevou-se e a paO2 mostrou-se maior no GM que no GX. Conclui-se que a infusão contínua de doses equipotentes de xilazina e medetomidina, durante anestesia geral inalatória, com isofluorano, em eqüinos, promove alterações cardiocirculatórias, respiratórias, térmicas e hemogasométricas discretas e equivalentes.

Eight horses under inhalant general anesthesia with isoflurane (1MAC) and continuous infusion of xylazine (0.35mg kg-1h-1) or medetomidine (3.5µg kg-1h-1) were evaluated for heart rate and rhythm, respiratory rate, arterial blood pressure, arterial blood gas analysis and temperature immediately before the beginning of the continuous infusion (T0) and in intervals of 10 minutes after the beginning of the continuous infusion (T10 to T60). Heart rate and temperature decreased and mean arterial pressure increased. PaCO2 (in GM) increased and GM showed a higher paO2 than GX. We conclude that equipotent doses of continuous infusion of medetomidine and xylazine during inhalant general anesthesia with isoflurane in horses promote slight and equivalent cardiocirculatory, respiratory, thermic and arterial blood gases changes.

Aspecto radiológico de heteroenxerto ósseo cortical fragmentado na reparação de falhas ósseas em coelhos / Radiologic aspects of fragmented cortical bone heterograft in bone defect repair in rabbits

Foi realizada falha segmentar de 6mm na região metafisária da tíbia de 12 coelhos,sendo utilizado enxerto ósseo cotical heterólogo fragmentado conservado em glicerina (98 por cento), para a sua reconstrução. O enxerto foi depositado no leito receptor e a integração deste foi avaliada radiologicamente por 20, 40 e 60 dias. Houve incorporação gradativa, em relação ao tempo, do enxerto no leito receptor, em 100 por cento dos casos. O uso de heteroenxerto ósseo cortical fragmentado e conservado em glicerina a 98 por cento promove a reparação das falhas ósseas, sem sinais de infecção ou rejeição, podendo ser utilizado como substituto ósseo na medicina veterinária...

It was realized a 6mm of segmental defect at the metaphyseal region of the tibia of 12 rabbits and fragmented heterolog cortical bone graft conserved in glycerin (98 percent) was used to the reconstruction. The graft was placed in the receptor bed and its integration was evaluated by radiological exam after 20, 40, and 60 days. There was gradative graft incorporation, by the time, in the receptor bed, in 100 percent of the cases. The use of fragmented cortical bone heterograf conserved in glycerin 98 percent promotes bone defect reparation without signs of infection and can be used like osseous substitute in the veterinary medicine...

Implante de tubo de silicone com e sem colágeno na regeneração de nervos em eqüinos / Implant of silicone tube with or without collagen in nerve regeneration of horses

A reconstituição cirúrgica de nervos em humanos, em cães e em diversos animais de laboratório é bastante utilizada e tem indicações clínicas e experimentais importantes. No entanto, em eqüinos há poucas informações sobre esta prática. Há relatos sobre a excessiva proliferação de tecido conjuntivo e a formação de neuromas à neurorrafia experimental, mesmo quando se utilizam tubos de silicone para condução do crescimento axonal. O presente estudo teve o objetivo de acrescentar informações sobre o processo de reparação de nervos periféricos em eqüinos por meio de implante de tubo de silicone preenchido ou não com colágeno. Para tanto, foram utilizados oito eqüinos, alocados em dois grupos: GI-13 semanas e GII-26 semanas de observação. Foi realizada secção dos nervos ulnares (NUs) e dos ramos cutâneos laterais dos 17º nervos torácicos (NTs), bilateralmente, seguindo tubulização, realizada em cada animal, alternando-se tubos de silicone vazios (TS), em um dos antímeros, ou preenchidos com solução de colágeno (TSC), no contralateral, deixando-se um espaço de 5mm entre os cotos. Nenhuma alteração foi encontrada ao exame do aparelho locomotor e as primeiras reações positivas ao teste de sensibilidade cutânea nos NUs e NTs com TS e TSC foram observadas a partir da 9ª semana, em ambos os grupos. Ao final do período de observação, verificou-se, macroscopicamente, que os nervos encontravam-se envolvidos por tecido conjuntivo e o interior da câmara estava preenchido por tecido de coloração esbranquiçada, de forma cilíndrica, interligando os cotos proximal e distal. Microscopicamente, constatou-se a presença de axônios mielinizados interligando os cotos, as células de Schwann e o processo de remielinização do coto distal, principalmente no TSC. Em ambos os grupos, não houve formação de neuromas e o processo inflamatório limitou-se às áreas perineurais. Concluiu-se que o implante de tubo de silicone conduz à regeneração de NUs e NTs de eqüinos...

Surgical nerve repair in humans, dogs and laboratory animals is widely utilized for important clinical and experimental purposes. However, in horses there is scarce information. Neuroma and excessive conjunctive proliferation are reported. The aim of the present study was to add information about the repair process utilizing implant of silicone tubes with or without collagen in nerve regeneration of horses. Eight horses were allocated in two groups: GI-13 weeks and GII-26 weeks of observation. A complete section of the ulnar nerves (UNs) and of the lateral cutaneous branch of the 17th thoracic nerves (TNs), bilaterally, was followed by repair with silicone tubes alternately filled with (STC) or without collagen (ST), with a 5mm gap between stumps. Clinical evaluation was performed weekly, by cutaneous sensitivity testing of the region innervated by the respective operated nerves and physical examination of the thoracic limbs. No alterations were found in the locomotor apparatus. The first positive reactions to the cutaneous test occurred at the 9th week post-surgery. At the end of the observation period it was verified, grossly, that the nerves were involved by conjunctive tissue and the lumen of the tube was filled by a whitish tissue, in a cylindrical shape. The ends nerves were interconnected. Microscopically, myelinated axons and Schwann cells were present in the gap between the stumps. In the GII the number of myelinated fibers was greater. The distal nerve showed remyelination process. There was no presence of neuroma and the inflammatory process was limited to the perineural areas. It was concluded that the implant of silicone tubes results in regeneration of UNs and TNs of horses without formation of neuromas and that de addition of collagen promotes an increase in the number of myelinated fibers.