Abstract Synonymous single nucleotide variants (sSNVs) do not alter the primary structure of a protein, thus it was previously accepted that they were neutral. Recently, several studies demonstrated their significance to a range of diseases. Still, variant prioritization strategies lack focus on sSNVs. Here, we identified 22,841 deleterious synonymous variants in 125,748 human exomes using two in silico predictors (SilVA and CADD). While 98.2% of synonymous variants are classified as neutral, 1.8% are predicted to be deleterious, yielding an average of 9.82 neutral and 0.18 deleterious sSNVs per exome. Further investigation of prediction features via Heterogeneous Ensemble Feature Selection revealed that impact on amino acid sequence and conservation carry the most weight for a deleterious prediction. Thirty nine detrimental sSNVs are not rare and are located on disease associated genes. Ten distinct putatively non-deleterious sSNVs are likely to be under positive selection in the North-Western European and East Asian populations. Taken together our analysis gives voice to the so-called silent mutations as we propose a robust framework for evaluating the deleteriousness of sSNVs in variant prioritization studies.

ABSTRACT: Thoracoscopy is replacing open lung biopsies because it is less invasive, usually the technique is done using three portals and intracorporeal suture technique. This study described the feasibility and efficacy of a novel pre tied loop ligature and to propose a thoracoscopic access strategy with two portals to perform lung biopsy in patients under 5 kg. Ten rabbits were positioned in dorsal recumbency. Total thoracoscopic lung biopsy was performed using a combined transdiaphragmatic approach and a right intercostal approach. A pre tied loop ligature was placed to perform a caudal lung lobe biopsy. Insufflation of the thoracic cavity was not performed. The total surgery time was 41.4 ± 14.5 min. The procedure was carried out free of complications that prevented slippage or tightening the knot or that made it come loose after the lung biopsy; there was no serious complication during the surgical procedure. The samples obtained averaged 1x0.64x0.45 cm (Length, Width, Depth) and were considered satisfactory according to the histopathologic evaluation. Thorax radiographs taken before and after the surgeries were compared and showed no pneumothorax or hemothorax. Necropsy confirmed no knot failure occurred at the biopsy site. The use of the novel pre tied loop ligature is a safe and effective technique, avoiding problems with the limited size of the thoracic cavity in small patients.

RESUMO: A toracoscopia está substituindo as biópsias pulmonares abertas por ser menos invasiva. Usualmente a técnica é feita utilizando três portais e ligaduras feitas intra corporalmente. O objetivo deste estudo foi descrever a viabilidade e eficácia de uma nova ligadura com alça pré-amarrada e propor uma estratégia de acesso toracoscópico com dois portais para realizar biópsia pulmonar em pacientes com menos de 5kg. Dez coelhos foram posicionados em decúbito dorsal. A biópsia pulmonar por toracoscopia total foi feita pela abordagem combinada transdiafragmática e abordagem intercostal direita. Uma nova ligadura pré-amarrada foi colocada para realizar uma biópsia do lobo pulmonar caudal. Não foi realizada insuflação da cavidade torácica. O tempo total de cirurgia foi de 41,4 ± 14,5 min. Os procedimentos foram realizados sem complicações que impedissem o escorregamento ou aperto do nó ou que o soltassem após a biópsia pulmonar; não houve nenhuma intercorrência grave durante o procedimento cirúrgico. As amostras coletadas tinham tamanho médio 1x0,64x0,45 cm e foram consideradas satisfatórias de acordo com a avaliação histopatológica. As radiografias de tórax feitas antes e depois das cirurgias foram comparadas e não mostraram pneumotórax ou hemotórax. A necropsia confirmou que não ocorreu falha do nó no local da biópsia. Conclui-se que uso da nova ligadura com alça pré-amarrada é uma técnica segura e eficaz, evitando problemas com o tamanho limitado da cavidade torácica em pacientes pequenos.

Resumo Há uma enorme disparidade entre os países de alta renda e outros em termos de acesso a dispositivos médicos cardíacos, como marca-passos e desfibriladores implantáveis. Os custos são uma das principais barreiras ao uso de dispositivos cardíacos nesses países. Existem iniciativas internacionais que visam reduzir essa disparidade, e o reuso de marca-passos tem sido discutido como uma possível alternativa. O conceito de reutilização de marca-passos não é novo; entretanto, estudos recentes têm se mostrado seguros, éticos e eficazes para aqueles que precisam de dispositivos eletrônicos cardíacos implantáveis e não tem como adquiri-los. Parte dos países de língua portuguesa, especialmente na África, precisam de uma resposta imediata que beneficie seus inúmeros pacientes que sofrem de arritmias tratáveis. marcapassos marca passos alternativa novo entretanto seguros adquirilos. adquirilos adquiri los. los adquiri-los portuguesa África tratáveis

Abstract There is a gap between high-income countries and others in terms of access to medical cardiac devices, such as pacemakers and implantable cardioverter defibrillators. Costs are one of the main barriers to the use of cardiac devices in these countries. There are international initiatives that aim to reduce the gap. The reuse of pacemakers has been discussed as a possible alternative to this problem. The concept of reusing pacemakers is not new; however, recent studies have proven to be safe, ethical, and effective for those who need cardiac implantable electronic devices and cannot afford them. Part of the Portuguese-speaking countries, especially in Africa, need an immediate response that benefits their countless patients who suffer from treatable arrhythmias. highincome high income defibrillators problem new however safe ethical them Portuguesespeaking Portuguese speaking Africa arrhythmias

Abstract Introduction Tinnitus is experienced by a significant part of the patients suffering from otosclerosis. Objective To assess the prevalence of tinnitus in otosclerosis, its main features, and the impact on the daily life. Methods Patients diagnosed with otosclerosis in 2019 in a tertiary hospital were enrolled in the study. Demographic data were retrieved and, besides a regular audiometric evaluation, the patients underwent acuphenometry to assess the psychoacoustic measurements (pitch and loudness), and the Tinnitus Handicap Inventory (THI). Results In total, 66 patients fulfilled the inclusion criteria, with a female predominance (63.6%; n = 42), and a mean age of 48.7 years. The mean air-bone gap was of 26.3 dB. A total of 72.7% complained of tinnitus; it was mostly unilateral, identified in the low frequencies, namely 500 Hz, with median loudness of 7.5 dB. The median score on the THI score was of 37; most patients had a mild handicap (33.3%, n = 16), followed by those with a severe handicap (22.9%; n = 11). The female gender had a statistically significant association with the presence of tinnitus. The THI scores were higher in middle-aged patients (age groups: 40 to 49 and 50 to 59 years), which was statistically significant. No correlation was found between audiometry results and the prevalence of tinnitus or score on the THI. On the other hand, high-pitched tinnitus, compared to low pitched-tinnitus, was associated with larger air-bone gaps. Conclusion The prevalence of tinnitus in our population was in line with the prevalences reported in the literature. It caused a catastrophic handicap in 22.9% of the patients. High-pitched tinnitus was associated with higher handicap. Nonetheless, the existence of tinnitus and its severity were not associated with the degree of hearing loss.

RESUMO: A haste intramedular bloqueada representa excelente opção para abordagem cirúrgica de fraturas em ossos longos na medicina veterinária. Todavia, falhas decorrentes principalmente de “folga” presente na interface da haste com os parafusos foram relatadas. O presente estudo teve como objetivo testar e comparar mecanicamente, por cargas de compressão axial, dois novos modelos de haste intramedular bloqueada de ângulo estável com orifícios roscados, sendo uma delas uniplanar e outra multiplanar com orifícios distais dispostos ortogonalmente. Vinte e um corpos de prova confeccionados em ácido polilático (PLA) foram utilizados para implantação das hastes intramedulares bloqueadas, divididos em três grupos: Haste bloqueada convencional (G1); haste bloqueada nova de ângulo estável com orifícios dispostos em único plano (G2); haste bloqueada nova de ângulo estável em dois planos, com o penúltimo orifício em 90 graus dos demais (G3). Utilizou-se ensaios biomecânicos por carga de compressão axial, cíclicos e destrutivos, para comparação entre eles. Todos mostraram deformação plástica nos parafusos após testes destrutivos, tanto proximais como distais, sendo mais intensa no grupo 1. Os grupos 2 e 3 das hastes de ângulo estável suportaram cargas superiores em relação ao G1 em todos os testes realizados (P<0.05). As hastes novas não diferiram estatisticamente entre si (P>0,05). A hipótese inicial de que os novos modelos proporcionariam maior estabilidade foi confirmada, entretanto não foi demonstrada diferenças entre eles. O sistema de bloqueio roscado dos parafusos nas hastes permitiu valores elevados de resistência nos testes realizados, mostrando-se efetivo e, potencialmente aplicável em situações clínicas reais.

ABSTRACT: The interlocking nail represents an excellent option as a surgical approach to treat fractures in long bones in veterinary medicine. However, failures were reported mainly due to a slack present in the interface of the rod with the screws. The present study tested and mechanically compare axial compression loads of two novel models of stable angle interlocking nails with threaded holes. Among the two models, one was uniplanar and the other was multiplanar with orthogonally arranged distal holes. Twenty-one specimens made of polylactic acid were used for the implantation of interlocking nail´s rods, divided into three groups: conventional interlocking nail (G1), novel interlocking nail with a stable angle with holes arranged in a single plane (G2), and novel interlocking nail with a stable angle in two planes, with the penultimate hole at 90 degrees from the others (G3). Biomechanical tests were performed using axial, cyclic, and destructive compression load for comparison between them. All the specimens showed plastic deformation in the screws after destructive tests, in both proximal and distal sides, being highly intense in G1. G2 and G3 of the stable angle rods supported higher loads than G1 in all tests performed (P<0.05). The novel stems did not differ statistically from each other (P>0.05). The initial hypothesis that the novel models would provide increased stability was confirmed; however, no differences were demonstrated between them. The screw locking system on the rods allowed high resistance values in the tests performed, proving to be effective and potentially applicable in real clinical situations.

ABSTRACT Purpose To evaluate clinical outcome following minimally invasive plate osteosynthesis (MIPO) associated with percutaneous transplantation of allogeneic adipose-derived mesenchymal stem cells (AD-MSC) at the tibial fracture site in dogs. Methods Thirty-six dogs presenting with nonarticular complete tibial fracture were included in this study. All fractures were treated by the same MIPO technique. The animals were divided in group 1 (n = 20) received a percutaneous application of 3 × 106 AD-MSC at the fracture site and group 2 (n = 16) did not receive any adjuvant treatment. Postoperative radiographic examinations were made at 15, 30, 60, 90 and 120 days. Results Fifty-eight percent of the patients were classified as skeletally immature. The median weight of the animals was 18.8 kg. The mean radiographic union time differed statistically between the AD-MSC group (28.5 days) and the control group (70.3 days). Sixty percent of dogs in group 1 and 56.25% of the group 2 were considered immature. Conclusions The use of allogeneic AD-MSC cell therapy and MIPO is a safe, viable and effective technique for promoting bone healing in nonarticular tibial fractures in dogs.

ABSTRACT Purpose To use a 3D printed poly (L-lactide) acid (PLLA) and hydroxyapatite (HA) composite as a bone substitute for reconstruction of a critical bone defect in the radius of rabbits. Methods A 1.5 cm ostectomy was performed in the radial diaphysis of 60 New Zealand white rabbits. The rabbits were divided into three groups according to surgical treatment of the bone defect (group I – control, group II – bone graft, group III – 3D PLLA). Each group was divided into four subgroups with different radiographic and histopathologic evaluation times (T1 – 15 days, T2 – 30 days, T3 – 60 days, T4 – 90 days). Results The implant group had greater clinically lameness (p = 0.02), edema (p = 0.007), pain (p = 0.04) and more complications at the surgical site (p = 0.03). Histologically, this group showed greater congestion (p = 0.04), hemorrhage (p = 0.04) and inflammation. Osteogenesis was microscopically similar between days (p = 0.54) and treatments (p = 0.17), even though radiographically, more effective bone healing occurred in the graft group (II), with more callus and bone bridge formation. Conclusions The customization of a 3D PLLA/HA scaffold was successful. However, in animals receiving the polymer-ceramic composite less bone callus and bone bridge was formed compared to the graft group.

RESUMO: Este estudo teve como objetivo avaliar radiograficamente as alterações no alinhamento da tíbia nos planos frontal e sagital em cães submetidos à osteossíntese minimamente invasiva com placa (MIPO) sem o auxílio de intensificadores de imagem. Foram incluídas e avaliadas radiografias de cães com fraturas completas da tíbia não articulares submetidos a MIPO, sem o auxílio de intensificador de imagem transoperatório e/ou associação de implantes. Os ângulos mecânicos tibiais (mMPTA, mMDTA, mCaPTA e mCrDTA) foram mensurados por três avaliadores. Os dados obtidos pela média de todas as avaliações foram comparados com resultados de estudos previamente publicados. Vinte e sete animais foram incluídos no estudo. A média e desvio padrão das alterações angulares foram os seguintes: mMPTA= 2,54° ± 3,10 (-1,1º a 8,7º); mMDTA= 0,03º ± 0,16 (-3,44º a 0,79º); mCaPTA= 37º ± 4,29 (-6,23º a 14,87º); e mCrDTA= 8,25 ° ± 5,53 (-0,2º a 17,28º). Houve uma correlação negativa entre “mCaPTA” e “mCrDTA”. A realização de MIPO em tíbia sem o uso de intensificadores de imagem e associação de implantes pode causar alterações angulares, o que pode levar a deformidades clinicamente relevantes após a cicatrização óssea.

ABSTRACT: This study assessed radiographically changes in tibial alignment in the frontal and sagittal planes in dogs that underwent minimally invasive plate osteosynthesis (MIPO) without the aid of image intensifiers. Radiographs of dogs with complete non-articular tibial fractures submitted to MIPO were included and evaluated, without the aid of a transoperative image intensifier and / or an association of implants. The tibial mechanical angles (mMPTA, mMDTA, mCaPTA and mCrDTA) were measured by three evaluators. The data obtained were compared with results from previously published studies. Twenty-seven animals were included in the study. The mean and standard deviation of the angular changes were as follows: mMPTA, 2.54° ± 3.10 (-1.1º to 8.7º); mMDTA, 0.03º ± 0.16 (-3.44º to 0.79º); mCaPTA, 37º ± 4.29 (-6.23º to 14.87º); and mCrDTA, 8.25° ± 5.53 (-0.2º to 17.28º). There was a negative correlation between “mCaPTA” and “mCrDTA”. MIPO of the tibia without using image intensifiers and implant association can potentially cause angular changes, which can lead to clinically relevant deformities after bone healing.

ABSTRACT Purpose To investigate the applicability of piezosurgery for cervical ventral slot (CVS), comparing it with the conventional technique of using high-speed burs for bone wear. Methods Thirty rabbits (Oryctolagus cuniculus) were divided into two treatment groups (T1 and T2) corresponding to CVS between C3-C4. In T1, the surgery was performed with piezoelectric apparatus, and in T2 with high-speed burs. The evaluated parameters were: duration of each stage of surgery, temperature variations during CVS, visibility of the surgical field, intra and postoperative complications, and anesthetic monitoring. At 14, 28, and 56 postoperative days, five animals from each treatment group were submitted for histopathological study of the surgical site. Results Compared with T2, T1 had more precise bone cut, and better visibility of the operative field, although it required longer total surgical time (p = 0.02) and triggered a greater number of intraoperative complications (p < 0.01), microscopic lesions in the spinal cord (p < 0.05), and transient neurological deficits in the postoperatively (p < 0.05). Conclusions It is necessary to perform surgical planning and have several tips of the piezoelectric instrument available for the safe use of the piezoelectric device in neurosurgery.

ABSTRACT Purpose To evaluate the biomechanical properties of a novel total hip replacement femoral stem. Methods Eight pairs of femurs from dog cadavers were used. The femurs were separated into different groups. A novel femoral stem with a convex proximal portion (Stem B) was biomechanically evaluated and compared to awell-known veterinary collared stem (Stem A). Femoral stems were inserted into the contralateral femurs from the same dog, forming 16 constructs. A flexo-compression load was applied on the axial axis of each sample. Maximum strength, deflection, stiffness, and energy absorption were analysed. Results Group B constructs showed significantly higher values (p ? 0.05) for the variables, except stiffness. The mean maximum strength was 1,347 ± 357 N for Group A and 1,805 ± 123 N for Group B (p ? 0.0069). The mean deflection was5.54 ± 2.63 mm for Group A and 10.03 ± 3.99 mm for Group B (p ? 0.0056). For the energy variable, the force was 6,203 ± 3,488 N/mm for Group A and 12,885 ± 5,056 N/mm for Group B (p ? 0.0054). Stem B had greater maximum strength, deflection, and energy. Conclusions The new stem was effective in neutralizing the impact of axial flexion-compression stresses during biomechanical tests in cadaveric models.

Introdução: O hematoma do recto abdominal é uma entidade rara e causa incomum de dor abdominal. Apesar de habitualmente benigno, é potencialmente fatal, sobretudo em doentes idosos e hipocoagulados. Pretendemos estudar a ocorrência e apresentação clínica desta entidade, focando a sua relação com a terapêutica hipocoagulante. Material e Métodos: Análise retrospetiva de uma série de casos observados num serviço de Medicina de um hospital terciário universitário e revisão da literatura. Foram incluídos doentes com diagnóstico de hematoma do recto abdominal de 2010 a 2018. Resultados: No período de estudo foram identificados 9 casos em 14 730 admissões, com idade média de 78 ± 9 anos, 6 do sexo feminino. Todos apresentaram hematoma do recto abdominal espontâneo. Quarenta e quatro por cento foram graves, segundo a classificação de Berná e a mortalidade foi de 11%. A apresentação mais frequente foi dor abdominal com queda de hemoglobina (média de 3,7 ± 1,8 g/dL) e confirmou-se o diagnóstico por tomografia computorizada. Todos os doentes estavam hipocoagulados. Os doentes sob dicumarínicos apresentaram INR supra-terapêutico (média 6,0 ± 2,6) e um factor precipitante (tosse), ao contrário dos doentes sob heparina de baixo peso molecular. Foi necessário reverter a anticoagulação em 6 casos. Cinco doentes tiveram alta hipocoagulados, sem recorrência de hematoma do recto abdominal. Discussão/Conclusão: O hematoma do recto abdominal deve ser considerado no diagnóstico diferencial de dor abdominal em doentes anticoagulados. Apesar de ser uma complicação rara da terapêutica hipocoagulante, pode resultar num desfecho fatal. A administração correcta da heparina de baixo peso molecular, o controlo rigoroso do INR e a elevada suspeita clínica poderão contribuir para uma menor morbi-mortalidade

Introduction: Rectus sheath hematoma is a rare entity and seldomly the cause of acute abdominal pain. Although mostly benign, it can be fatal especially in older anticoagulated patients. We aim to study the occurrence and clinical presentation of this entity, focusing on its relationship with anticoagulant therapy. Material and Methods: Retrospective case series of patients admitted to an Internal Medicine Unit of a tertiary university hospital and revision of the literature. Patients with the diagnosis of rectus sheath hematoma from 2010 to 2018 were included. Results: In the study period, there were nine cases of rectus sheath hematoma in 14 730 admissions, with median age of 78 ± 9 years, of which 6 were women. All patients presented with spontaneous rectus sheath hematoma. Forty-four percent were severe, according to the Classification of Berná, and the mortality rate was 11%. Abdominal pain and a fall in the haemoglobin level (mean of 3.7 ± 1.8 g/dL) were the most common clinical manifestations and diagnosis was confirmed by computed tomography. All patients were under anticoagulants. Those treated with dicoumarins had a supratherapeutic INR (mean 6.0 ± 2.6) and a precipitating factor (cough), contrary to those under low-molecular weight heparin. Anticoagulation was reversed in 6 cases. Five patients were discharged under anticoagulants and had no rectus sheath hematoma recurrence. Discussion/Conclusion: Rectus sheath hematoma must be considered in the differential diagnosis of patients with abdominal pain under anticoagulant therapy. Although a rare complication, it can have a fatal outcome. Efforts to guarantee a correct administration of low-molecular weight heparin, rigorous INR control and high clinical suspicion with early diagnosis are necessary to avoid it.

Abstract Purpose To evaluate the use of demineralized bone matrix of caprine origin in experimental bone defects of the tibia in New Zealand rabbits. Methods Fragments of the tibia diaphysis were collected aseptically from clinically healthy goats. The bones were sectioned into 1 cm fragments and stored at -20°C for subsequent hydrochloric acid (HCL) demineralization. A 70 mg portion of DBMc was used to fill the experimental bone defects. Twenty-four female adult New Zealand rabbits were divided into 2 groups: the MG (matrix group, left tibia) and CG (control group, right tibia). Additionally, they were separated into 4 groups with 6 animals, according to the period of analysis (15, 30, 60 and 90 days postoperatively). Using microCT, volumetric parameters were evaluated: bone volume, relationship between bone volume and total volume, bone surface area, relationship between bone surface area and total volume, number of trabeculae, trabecular thickness and trabecular separation. Results There was a statistically significant difference (P<0.05) between groups considering bone volume (BV) and bone:total volume (BV/TV), on 15, 30 and 90 days postoperatively. Control group showed a statistically significant superiority (P < 0.05) considering the mean of the variables bone surface (BS), number of trabeculae (Tb.N) and between bone surface and total volume (BS/TV) at 15 and 90 days. Conclusions Caprine demineralized bone matrix was safe and tolerable. No signs of material rejection were seen macroscopically. It is an alternative for the treatment of bone defects when autologous graft is not available or in insufficient quantities.

Abstract Purpose: To evaluate clinically dogs that underwent tibial tuberosity advancement (TTA) six months previously. Methods: Dogs of various breeds, gender, weight, and age that had CCL rupture and underwent TTA for treatment were included in this study. Parapatellar arthrotomy was performed in all patients to assess the joint for a ruptured ligament and meniscal injury before the TTA. The appropriate cage for the TTA was chosen with planning surgery. The surgical procedure was performed according to the literature, using a modified Maquet technique. Six months after surgery, lameness during walking; muscular atrophy; crepitation, cranial drawer and tibial compression tests and quality of life based on owner’s evaluation were assessed. Results: Postoperative complications were observed in only one knee (4.76%), with a surgical site seroma. The mean lameness score at walking was 0.29 (± 0.64). The mean score regarding muscular atrophy was 0.95 (± 1.56). The mean score of the cranial drawer test, in a range from 0 to 5, was 1.52 (± 1.54). The owners rated the dog’s quality of life as excellent in 44%, good in 30%, and moderate in 17%. Conclusion: This clinical study supports the affirmation that patients who undergo TTA for treatment of CCL rupture have an acceptable response.

RESUMO: O objetivo deste trabalho foi descrever e analisar a funcionalidade da configuração tie-in na ostessíntese de tíbia em cães. Foram usados 21 cães com fratura de tíbia da rotina clinica do hospital veterinário. Somente a primeira cortical (Sis) foi perfurada, em seguida a broca foi inclinada a 45o e projetada no mesmo orifício no sentido distal do osso. foram confeccionados outros orifícios com auxílio de furadeira de baixa rotação e broca cujo diâmetro era menor que o implante escolhido. Os procedimentos radiográficos foram realizados no momento do atendimento clínico, no período pós-operatório imediato e aos 30, 60, 90 e 120 dias pós-cirúrgico. Não houve nenhuma complicação transoperatória, tão pouco deiscência de sutura até o momento da retirada dos pontos de cútis. A formação parcial de calo ósseo foi evidenciada em 20 cães com tempo médio de 76 dias. Três cães obtiveram consolidação óssea em 35 dias, nove aos 60 dias, três aos 90 dias e cinco aos 120 dias de pós-operatório. Dinamizações foram realizadas em nove animais. Após a confirmação radiográfica de consolidação óssea, os implantes dos 20 animais foram totalmente removidos. Concluiu-se que o acesso cirúrgico ao canal medular da tíbia canina por meio de orifício na face medial proximal, junto à tuberosidade da tíbia, permite a inserção do PIM sem risco de lesões articulares e periarticulares no joelho.

ABSTRACT: The aim of this study was to describe and analyse the adaptability and functionality of tie-in configuration in tibial osteosynthesis in dogs. Twenty dogs with tibial fracture were included in this study. An orifice was made on the proximal tibial fragment, on the medial side, close to the tibial crest. The drill piece was angled at 45º and projected into the same orifice on the distal sense of the bone. Others orifices made with the aid of a low rotation drill and drill piece with diameter smaller than the chosen implant. After 10 days post-operative, the animals were evaluated. X-ray analysis was performed at the time of clinical examination; immediate post-operative period; and at 30, 60, 90, and 120 days post-surgery. A questionnaire was given to the owners to provide details on the post-operative period. There were no trans-operative complications or suture dehiscence up to the day of suture removal. Partial development of bone callus was observed in 20 dogs within a mean period of 76 days. Three animals showed bone consolidation within 35 days, nine by 60 days, three by 90 days, and 5 by 120 days post-operative. Dynamization was carried out in 9 animals. The surgical access to the tibial medullary canal through the orifice at the proximal medial face, by the tibial tuberosity, enables the insertion of IMP without risks to articular and peri-articular lesions in the knee in dogs.

Electrochemical aptasensor arrays have been recently used to detect several proteins reported as disease biomarkers, mainly due to their known advantages, that include high affinity, sensitivity, specificity and low costs, among others. This study describes the development of a label-free electrochemical multi-aptasensor array, for the simultaneous detection of human osteopontin (OPN), using two specific aptamers. To enable multiplexed protein assay, RNA and DNA aptamers were immobilized in the dual-screen-printed gold working electrodes via streptavidin-biotin interaction, and using the [Fe(CN)6]3-/4- as the redox probe for cyclic voltammetry measurements. The multi-aptasensor array herein developed exhibited a good response and selectivity to detect human OPN in the presence of other interfering proteins. Considering these preliminary results, the DNA/RNA dual aptasensor array could potentially be used as an analytical tool for the specific detection of human OPN, and for cancer diagnosis overall.