Geometrical optimization of a short-stem hip implant for the reduction of proximal stress shielding (ottimizzazione geometrica di uno stelo corto di protesi d?anca per la riduzione della schermatura del carico prossimale)

Nowadays the number of hip joints arthroplasty operations continues to increase because the elderly population is growing. Moreover, the global life expectancy is increasing and people adopt a more active way of life. For this reasons, the demand of implant revision operations is becoming more frequent. The operation procedure includes the surgical removal of the old implant and its substitution with a new one. Every time a new implant is inserted, it generates an alteration in the internal femur strain distribution, jeopardizing the remodeling process with the possibility of bone tissue loss. This is of major concern, particularly in the proximal Gruen zones, which are considered critical for implant stability and longevity. Today, different implant designs exist in the market; however there is not a clear understanding of which are the best implant design parameters to achieve mechanical optimal conditions. The aim of the study is to investigate the stress shielding effect generated by different implant design parameters on proximal femur, evaluating which ranges of those parameters lead to the most physiological conditions.

Proliferation, differentiation and gene expression of osteoblasts in boron-containing associated with dexamethasone deliver from mesoporous bioactive glass scaffolds

Boron is one of the trace elements in the human body which plays an important role in bone growth. Porous mesopore bioactive glass (MBG) scaffolds are proposed as potential bone regeneration materials due to their excellent bioactivity and drug-delivery ability. The aims of the present study were to develop boron-containing MBG (B-MBG) scaffolds by sol-gel method and to evaluate the effect of boron on the physiochemistry of B-MBG scaffolds and the response of osteoblasts to these scaffolds. Furthermore, the effect of dexamethasone (DEX) delivery in B-MBG scaffold system was investigated on the proliferation, differentiation and bone-related gene expression of osteoblasts. The composition, microstructure and mesopore properties (specific surface area, nano-pore volume and nano-pore distribution) of B-MBG scaffolds have been characterized. The effect of boron contents and large-pore porosity on the loading and release of DEX in B-MBG scaffolds were also investigated. The results have shown that the incorporation of boron into MBG scaffolds slightly decreases the specific surface area and pore volume, but maintains well-ordered mesopore structure and high surface area and nano-pore volume compared to non-mesopore bioactive glass. Boron contents in MBG scaffolds did not influence the nano-pore size distribution or the loading and release of DEX. B-MBG scaffolds have the ability to maintain a sustained release of DEX in a long-term span. Incorporating boron into MBG glass scaffolds led to a controllable release of boron ions and significantly improved the proliferation and bone-related gene expression (Col I and Runx2) of osteoblasts. Furthermore, the sustained release of DEX from B-MBG scaffolds significantly enhanced alkaline phosphatase (ALP) activity and gene expressions (Col I, Runx2, ALP and BSP) of osteoblasts. These results suggest that boron plays an important role in enhancing osteoblast proliferation in B-MBG scaffold system and DEX-loaded B-MBG scaffolds show great potential as a release system to enhance osteogenic property for bone tissue engineering application.

Lateral ridge augmentation using a PCL-TCP scaffold in a clinically relevant but challenging micropig model

In implant dentistry, there is a need for synthetic bone substitute blocks to support ridge augmentation in situations where large bone volumes are missing. Polycaprolactone-based scaffolds demonstrated excellent results in bone tissue engineering applications. The use of customized polycaprolactone-tricalcium phosphate (PCL-TCP) displayed promising results from recent rat femur and rabbit calvaria studies. However, data from clinically representative models in larger animals do not exist.

Development of Vapor Deposited Silica Sol-Gel Particles for a Bioactive Materials System to Direct Osteoblast Behavior

Tissue engineering and regenerative medicine have emerged in an effort to generate replacement tissues capable of restoring native tissue structure and function, but because of the complexity of biologic system, this has proven to be much harder than originally anticipated. Silica based bioactive glasses are popular as biomaterials because of their ability to enhance osteogenesis and angiogenesis. Sol-gel processing methods are popular in generating these materials because it offers: 1) mild processing conditions; 2) easily controlled structure and composition; 3) the ability to incorporate biological molecules; and 4) inherent biocompatibility. The goal of this work was to develop a bioactive vaporization system for the deposition of silica sol-gel particles as a means to modify the material properties of a substrate at the nano- and micro- level to better mimic the instructive conditions of native bone tissue, promoting appropriate osteoblast attachment, proliferation, and differentiation as a means for supporting bone tissue regeneration. The size distribution, morphology and degradation behavior of the vapor deposited sol-gel particles developed here were found to be dependent upon formulation (H2O:TMOS, pH, Ca/P incorporation) and manufacturing (substrate surface character, deposition time). Additionally, deposition of these particles onto substrates can be used to modify overall substrate properties including hydrophobicity, roughness, and topography. Deposition of Ca/P sol particles induced apatite-like mineral formation on both two- and three-dimensional materials when exposed to body fluids. Gene expression analysis suggests that Ca/P sol particles induce upregulation osteoblast gene expression (Runx2, OPN, OCN) in preosteoblasts during early culture time points. Upon further modification-specifically increasing particle stability-these Ca/P sol particles possess the potential to serve as a simple and unique means to modify biomaterial surface properties as a means to direct osteoblast differentiation.

Immunohistochemical localization of RANK, RANKL and OPG in healthy and arthritic canine elbow joints

OBJECTIVE: To determine if the receptor activator of nuclear factor-kappaB-receptor activator of nuclear factor-kappaB ligand-osteoprotegerin (RANK-RANKL-OPG) system is active in bone remodeling in dogs and, if so, whether differences in expression of these mediators occur in healthy and arthritic joints. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Fragmented processus coronoidei (n=20) were surgically removed from dogs with elbow arthritis and 5 corresponding healthy samples from dogs euthanatized for reasons other than elbow joint disease. METHODS: Bright-field immunohistochemistry and high-resolution fluorescence microscopy were used to investigate the distribution of RANK, RANKL, and OPG in healthy and arthritic joints. RESULTS: All 3 molecules were identified by immunostaining of canine bone tissue. In elbow dysplasia, the number of RANK-positive osteoclasts was increased. In their vicinity, cells expressing RANKL, a mediator of osteoclast activation, were abundant whereas the number of osteoblasts having the potential to limit osteoclastogenesis and bone resorption via OPG was few. CONCLUSIONS: The RANK-RANKL-OPG system is active in bone remodeling in dogs. In elbow dysplasia, a surplus of molecules promoting osteoclastogenesis was evident and is indicative of an imbalance between the mediators regulating bone resorption and bone formation. Both OPG and neutralizing antibodies against RANKL have the potential to counterbalance bone resorption. CLINICAL RELEVANCE: Therapeutic use of neutralizing antibodies against RANKL to inhibit osteoclast activation warrants further investigation.

BMP-2 liberated from biomimetic implant coatings induces and sustains direct ossification in an ectopic rat model

INTRODUCTION: Using a rat model, we evaluated the kinetics and histomorphometry of ectopic bone formation in association with biomimetic implant coatings containing BMP-2. MATERIALS AND METHODS: One experimental and three control groups were set up: titanium-alloy discs coated with a biomimetically co-precipitated layer of calcium phosphate and BMP-2 [1.7 microg per disc (incorporated-BMP group)]; uncoated discs (control); discs biomimetically coated with a layer of calcium phosphate alone (control); and discs biomimetically coated with a layer of calcium phosphate bearing superficially adsorbed BMP-2 [0.98 microg per disc (control)]. Discs (n = 6 per group) were implanted subcutaneously in rats and retrieved at 7-day intervals over a period of 5 weeks for kinetic, histomorphometrical, morphological and histochemical analyses. RESULTS: In the incorporated-BMP-2 group, osteogenic activity was first observed 2 weeks after implantation and thereafter continued unabated until the end of the monitoring period. The net weekly rates of bone formation per disc were 5.8 mm3 at 2 weeks and 3.64 mm3 at 5 weeks. The total volumes of bone formed per disc at these junctures were 5.8 mm3 and 10.3 mm3, respectively. Bone tissue, which was formed by a direct ossification mechanism, was deposited at distances of up to 340 microm from the implant surfaces. The biomimetic coatings were degraded gradually, initially by foreign body giant cells alone and then also by osteoclasts. Forty percent of the coating material (and thus presumably of the incorporated BMP-2) remained at the end of the monitoring period. Hence, 60% of the incorporated BMP-2 had been released. At this 5-week juncture, no bone tissue was associated with any of the control implants. CONCLUSION: BMP-2 incorporated into biomimetic calcium phosphate coatings is capable not only of inducing bone formation at an ectopic site in vivo but also of doing so with a very high potency at a low pharmacological level, and of sustaining this activity for a considerable period of time. The sustainment of osteogenic activity is of great clinical importance for the osseointegration of dental and orthopedic implants.

Osteoinductive implants: the mise-en-scène for drug-bearing biomimetic coatings

In orthopaedic and dental implantology, novel tools and techniques are being sought to improve the regeneration of bone tissue. Numerous attempts have been made to enhance the osteoconductivity of titanium prostheses, including modifications in their surface properties and coating with layers of calcium phosphate. The technique whereby such layers are produced has recently undergone a revolutionary change, which has had profound consequences for their potential to serve as drug-carrier systems. Hitherto, calcium phosphate layers were deposited upon the surfaces of metal implants under highly unphysiological physical conditions, which precluded the incorporation of proteinaceous osteoinductive drugs. These agents could only be adsorbed, superficially, upon preformed layers. Such superficially adsorbed molecules are released too rapidly within a biological milieu to be effective in their osteoinductive capacity. Now, it is possible to deposit calcium phosphate layers under physiological conditions of temperature and pH by the so-called biomimetic process, during which bioactive agents can be coprecipitated. Since these molecules are integrated into the inorganic latticework, they are released gradually in vivo as the layer undergoes degradation. This feature enhances the capacity of these coatings to act as a carrier system for osteogenic agents.

Multiple osteosclerotic lesions in an Iron Age skull from Switzerland (320 - 250 BC) - An unusual case

The single Hochdorf burial was found in 1887 during construction work in the Canton of Lucerne, Switzerland. It dates from between 320 and 250 BC. The calvarium, the left half of the pelvis and the left femur were preserved. The finding shows an unusual bony alteration of the skull. The aim of this study was to obtain a differential diagnosis and to examine the skull using various methods. Sex and age were determined anthropologically. Radiological examinations were performed with plain X-ray imaging and a multislice computed tomography (CT) scanner. For histological analysis, samples of the lesion were taken. The pathological processing included staining after fixation, decalcification, and paraffin embedding. Hard-cut sections were also prepared. The individual was female. The age at death was between 30 and 50 years. There is an intensely calcified bone proliferation at the right side of the os frontalis. Plain X-ray and CT imaging showed a large sclerotic lesion in the area of the right temple with a partly bulging appearance. The inner boundary of the lesion shows multi-edged irregularities. There is a diffuse thickening of the right side. In the left skull vault, there is a mix of sclerotic areas and areas which appear to be normal with a clear differentiation between tabula interna, diploë and tabula externa. Histology showed mature organised bone tissue. Radiological and histological findings favour a benign condition. Differential diagnoses comprise osteomas which may occur, for example, in the setting of hereditary adenomatous polyposis coli related to Gardner syndrome.

Periodontal and peri-implant wound healing following laser therapy

Laser irradiation has numerous favorable characteristics, such as ablation or vaporization, hemostasis, biostimulation (photobiomodulation) and microbial inhibition and destruction, which induce various beneficial therapeutic effects and biological responses. Therefore, the use of lasers is considered effective and suitable for treating a variety of inflammatory and infectious oral conditions. The CO2 , neodymium-doped yttrium-aluminium-garnet (Nd:YAG) and diode lasers have mainly been used for periodontal soft-tissue management. With development of the erbium-doped yttrium-aluminium-garnet (Er:YAG) and erbium, chromium-doped yttrium-scandium-gallium-garnet (Er,Cr:YSGG) lasers, which can be applied not only on soft tissues but also on dental hard tissues, the application of lasers dramatically expanded from periodontal soft-tissue management to hard-tissue treatment. Currently, various periodontal tissues (such as gingiva, tooth roots and bone tissue), as well as titanium implant surfaces, can be treated with lasers, and a variety of dental laser systems are being employed for the management of periodontal and peri-implant diseases. In periodontics, mechanical therapy has conventionally been the mainstream of treatment; however, complete bacterial eradication and/or optimal wound healing may not be necessarily achieved with conventional mechanical therapy alone. Consequently, in addition to chemotherapy consisting of antibiotics and anti-inflammatory agents, phototherapy using lasers and light-emitting diodes has been gradually integrated with mechanical therapy to enhance subsequent wound healing by achieving thorough debridement, decontamination and tissue stimulation. With increasing evidence of benefits, therapies with low- and high-level lasers play an important role in wound healing/tissue regeneration in the treatment of periodontal and peri-implant diseases. This article discusses the outcomes of laser therapy in soft-tissue management, periodontal nonsurgical and surgical treatment, osseous surgery and peri-implant treatment, focusing on postoperative wound healing of periodontal and peri-implant tissues, based on scientific evidence from currently available basic and clinical studies, as well as on case reports.

Análisis de micromovilidad en vástagos femorales mediante ensayo dinámico de oscilaciones libres

La artroplastia de cadera se considera uno de los mayores avances quirúrgicos de la Medicina. La aplicación de esta técnica de Traumatología se ha incrementado notablemente en los últimos anos, a causa principalmente del progresivo incremento de la esperanza de vida. En efecto, con la edad aumentan los problemas de artrosis y osteoporosis, enfermedades típicas de las articulaciones y de los huesos que requieren en muchos casos la sustitución protésica total o parcial de la articulación. El buen comportamiento funcional de una prótesis depende en gran medida de la estabilidad primaria, es decir, el correcto anclaje de la prótesis en el momento de su implantación. Las prótesis no cementadas basan su éxito a largo plazo en la osteointegración que tiene lugar entre el material protésico y el tejido óseo, y para lograrla es imprescindible conseguir unas buenas condiciones de estabilidad primaria. El aflojamiento aséptico es la principal causa de fallo de artroplastia total de cadera. Este es un fenómeno en el que, debido a complejas interacciones de factores mecánicos y biológicos, se producen movimientos relativos que comprometen la funcionalidad del implante. La minimización de los correspondientes danos depende en gran medida de la detección precoz del aflojamiento. Para lograr la detección temprana del aflojamiento aséptico del vástago femoral se han ensayado diferentes técnicas, tanto in vivo como in vitro: análisis numéricos y técnicas experimentales basadas en sensores de movimientos provocados por cargas transmitidas natural o artificialmente, tales como impactos o vibraciones de distintas frecuencias. Los montajes y procedimientos aplicados son heterogéneos y, en muchas ocasiones, complejos y costosos, no existiendo acuerdo sobre una técnica simple y eficaz de aplicación general. Asimismo, en la normativa vigente que regula las condiciones que debe cumplir una prótesis previamente a su comercialización, no hay ningún apartado referido específicamente a la evaluación de la bondad del diseño del vástago femoral con respecto a la estabilidad primaria. El objetivo de esta tesis es desarrollar una metodología para el análisis, in vitro, de la estabilidad de un vástago femoral implantado, a fin de poder evaluar las técnicas de implantación y los diferentes diseños de prótesis previamente a su oferta en el mercado. Además se plantea como requisito fundamental que el método desarrollado sea sencillo, reversible, repetible, no destructivo, con control riguroso de parámetros (condiciones de contorno de cargas y desplazamientos) y con un sistema de registro e interpretación de resultados rápido, fiable y asequible. Como paso previo, se ha realizado un análisis cualitativo del problema de contacto en la interfaz hueso-vástago aplicando una técnica optomecánica del campo continuo (fotoelasticidad). Para ello se han fabricado tres modelos en 2D del conjunto hueso-vástago, simulando tres tipos de contactos en la interfaz: contacto sin adherencia y con holgura, contacto sin adherencia y sin holgura, y contacto con adherencia y homogéneo. Aplicando la misma carga a cada modelo, y empleando la técnica de congelación de tensiones, se han visualizado los correspondientes estados tensionales, siendo estos más severos en el modelo de unión sin adherencia, como cabía esperar. En todo caso, los resultados son ilustrativos de la complejidad del problema de contacto y confirman la conveniencia y necesidad de la vía experimental para el estudio del problema. Seguidamente se ha planteado un ensayo dinámico de oscilaciones libres con instrumentación de sensores resistivos tipo galga extensométrica. Las muestras de ensayo han sido huesos fémur en todas sus posibles variantes: modelos simplificados, hueso sintético normalizado y hueso de cadáver, seco y fresco. Se ha diseñado un sistema de empotramiento del extremo distal de la muestra (fémur) con control riguroso de las condiciones de anclaje. La oscilación libre de la muestra se ha obtenido mediante la liberación instantánea de una carga estética determinada y aplicada previamente, bien con una maquina de ensayo o bien por gravedad. Cada muestra se ha instrumentado con galgas extensométricas convencionales cuya señal se ha registrado con un equipo dinámico comercial. Se ha aplicado un procedimiento de tratamiento de señal para acotar, filtrar y presentar las respuestas de los sensores en el dominio del tiempo y de la frecuencia. La interpretación de resultados es de tipo comparativo: se aplica el ensayo a una muestra de fémur Intacto que se toma de referencia, y a continuación se repite el ensayo sobre la misma muestra con una prótesis implantada; la comparación de resultados permite establecer conclusiones inmediatas sobre los efectos de la implantación de la prótesis. La implantación ha sido realizada por un cirujano traumatólogo utilizando las mismas técnicas e instrumental empleadas en el quirófano durante la práctica clínica real, y se ha trabajado con tres vástagos femorales comerciales. Con los resultados en el dominio del tiempo y de la frecuencia de las distintas aplicaciones se han establecido conclusiones sobre los siguientes aspectos: Viabilidad de los distintos tipos de muestras sintéticas: modelos simplificados y fémur sintético normalizado. Repetibilidad, linealidad y reversibilidad del ensayo. Congruencia de resultados con los valores teóricos deducidos de la teoría de oscilaciones libres de barras. Efectos de la implantación de tallos femorales en la amplitud de las oscilaciones, amortiguamiento y frecuencias de oscilación. Detección de armónicos asociados a la micromovilidad. La metodología se ha demostrado apta para ser incorporada a la normativa de prótesis, es de aplicación universal y abre vías para el análisis de la detección y caracterización de la micromovilidad de una prótesis frente a las cargas de servicio. ABSTRACT Total hip arthroplasty is considered as one of the greatest surgical advances in medicine. The application of this technique on Traumatology has increased significantly in recent years, mainly due to the progressive increase in life expectancy. In fact, advanced age increases osteoarthritis and osteoporosis problems, which are typical diseases of joints and bones, and in many cases require full or partial prosthetic replacement on the joint. Right functional behavior of prosthesis is highly dependent on the primary stability; this means it depends on the correct anchoring of the prosthesis at the time of implantation. Uncemented prosthesis base their long-term success on the quality of osseointegration that takes place between the prosthetic material and bone tissue, and to achieve this good primary stability conditions is mandatory. Aseptic loosening is the main cause of failure in total hip arthroplasty. This is a phenomenon in which relative movements occur, due to complex interactions of mechanical and biological factors, and these micromovements put the implant functionality at risk. To minimize possible damage, it greatly depends on the early detection of loosening. For this purpose, various techniques have been tested both in vivo and in vitro: numerical analysis and experimental techniques based on sensors for movements caused by naturally or artificially transmitted loads, such as impacts or vibrations at different frequencies. The assemblies and methods applied are heterogeneous and, in many cases, they are complex and expensive, with no agreement on the use of a simple and effective technique for general purposes. Likewise, in current regulations for governing the conditions to be fulfilled by the prosthesis before going to market, there is no specific section related to the evaluation of the femoral stem design in relation to primary stability. The aim of this thesis is to develop a in vitro methodology for analyzing the stability of an implanted femoral stem, in order to assess the implantation techniques and the different prosthesis designs prior to its offer in the market. We also propose as a fundamental requirement that the developed testing method should be simple, reversible, repeatable, non-destructive, with close monitoring of parameters (boundary conditions of loads and displacements) and with the availability of a register system to record and interpret results in a fast, reliable and affordable manner. As a preliminary step, we have performed a qualitative analysis of the contact problems in the bone-stem interface, through the application of a continuous field optomechanical technique (photoelasticity). For this proposal three 2D models of bone–stem set, has been built simulating three interface contact types: loosened an unbounded contact, unbounded and fixed contact, and bounded homogeneous contact. By means of applying the same load to each model, and using the stress freezing technique, it has displayed the corresponding stress states, being more severe as expected, in the unbounded union model. In any case, the results clearly show the complexity of the interface contact problem, and they confirm the need for experimental studies about this problem. Afterward a free oscillation dynamic test has been done using resistive strain gauge sensors. Test samples have been femur bones in all possible variants: simplified models, standardized synthetic bone, and dry and cool cadaveric bones. An embedding system at the distal end of the sample with strong control of the anchoring conditions has been designed. The free oscillation of the sample has been obtained by the instantaneous release of a static load, which was previously determined and applied to the sample through a testing machine or using the gravity force. Each sample was equipped with conventional strain gauges whose signal is registered with a marketed dynamic equipment. Then, it has applied a signal processing procedure to delimit, filter and present the time and frequency response signals from the sensors. Results are interpreted by comparing different trials: the test is applied to an intact femur sample which is taken as a reference, and then this test is repeated over the same sample with an implanted prosthesis. From comparison between results, immediate conclusions about the effects of the implantation of the prosthesis can be obtained. It must be said that the implementation has been made by an expert orthopedic surgeon using the same techniques and instruments as those used in clinical surgery. He has worked with three commercial femoral stems. From the results obtained in the time and frequency domains for the different applications the following conclusions have been established: Feasibility of the different types of synthetic samples: simplified models and standardized synthetic femur. Repeatability, linearity and reversibility of the testing method. Consistency of results with theoretical values deduced from the bars free oscillations theory. Effects of introduction of femoral stems in the amplitude, damping and frequencies of oscillations Detection of micromobility associated harmonics. This methodology has been proved suitable to be included in the standardization process of arthroplasty prosthesis, it is universally applicable and it allows establishing new methods for the analysis, detection and characterization of prosthesis micromobility due to functional loads.

Desenvolvimento de hidroxiapatita contendo prata via precipitação e imersão: avaliação do efeito antimicrobiano

A utilização de substitutos ósseos para recuperação da função perdida é uma constante busca dentro da área médica. Por isso os biomateriais têm recebido uma atenção muito grande por parte da comunidade científica, dentre eles os materiais a base de fosfato de cálcio. A hidroxiapatita, Ca10 (PO4)6 (OH) 2, tem sido muito estudada, pois além de representar a constituição da massa dos ossos naturais e dentes em 30 a 70%, possui propriedades de bioatividade e osteocondutividade, favorecendo e auxiliando o crescimento do tecido ósseo. Em contrapartida, infecções bacterianas podem surgir após o implante ocasionando a perda da funcionalidade a curto e médio prazo. Várias alternativas estão sendo testadas, geralmente associadas ao uso de antibióticos convencionais incorporados aos biomateriais. Uma alternativa a tais antibióticos seria a utilização de metais que possuem propriedades antibacterianas. A prata (Ag) é conhecida como um metal bactericida e por isso ganhou lugar de destaque dentre os estudos como um aliado importante no controle das infecções pós-cirúrgicas. Este trabalho teve como objetivo sintetizar, caracterizar e avaliar o efeito antimicrobiano da adição de íons de prata em hidroxiapatita. Foram obtidos pós de hidroxiapatita contendo prata (HAAg), nas concentrações de 0,1M; 0,01M e 0,001M pelo método de precipitação em temperatura ambiente e por imersão do pó de hidroxiapatita em soluções aquosas. As fases cristalinas e os grupamentos iônicos foram analisados para cada condição por técnicas de difração de raios X (DRX) e espectroscopia no infravermelho (IV) respectivamente. As informações sobre a morfologia e identificação de elementos químicos foi realizado pela técnica de microscopia eletrônica de varredura com espectroscopia de energia dispersiva (MEV EDS). As avaliações antimicrobianas foram realizadas por ensaios qualitativos e quantitativos, o ensaio qualitativo utilizou o teste de halo de difusão em disco para Staphylococcus aureus e Escherichia coli e o ensaio quantitativo utilizou contagem de bactérias para as cepas de Staphylococcus aureus. Os resultados de DRX e IV indicaram que independentemente do método de obtenção da HAAg foi possível observar a presença de prata metálica caracterizada pelos picos em 2θ=38,1º e 44,3º nas amostras HAAg0,1Im, HAAg0,1Pr e HAAg0,01Pr. Observou-se também a presença de AgO, correspondente ao pico em 2θ=37,5º nas amostras de HAAg0,01Pr e HAAg0,001Pr. Nos espectros de IV estão presentes as bandas que caracterizam a fase HA, referentes aos grupamentos PO43-, OH- e CO32-. Analisados em conjunto os ensaios qualitativos e quantitativos, as amostras HAAg0,01Im e HAAg0,001Im sintetizadas por imersão indicaram os melhores resultados para o ensaio de disco difusão, por apresentarem formação de halo inibição do crescimento bacteriano para a bactéria S. aureus. Para os ensaios quantitativos as amostras obtidas por precipitação com concentrações 0,1M e 0,01M de prata apresentaram melhor resultado por inibirem o crescimento bacteriano para as cepas S. aureus.

Participação do nicho endosteal na regulação da hemopoese de camundongos submetidos à desnutrição proteica

O nicho endosteal da medula óssea abriga as células-tronco hemopoéticas (CTH) em quiescência/autorrenovação. As CTH podem ser classificadas em dois grupos: células que reconstituem a hemopoese em longo prazo (LT-CTH) e curto prazo (CT-CTH). Investigamos, neste trabalho, os efeitos da desnutrição proteica (DP) no tecido ósseo e a participação do nicho endosteal na sinalização osteoblasto-CTH. Para tanto, utilizamos camundongos submetidos à DP induzida pelo consumo de ração hipoproteica. Os animais desnutridos apresentaram pancitopenia e diminuição nas concentrações de proteínas séricas e albumina. Quantificamos as CTH por citometria de fluxo e verificamos que os desnutridos apresentaram menor porcentagem de LT-CTH, CT-CTH e de progenitores multipotentes (PMP). Avaliamos a expressão das proteínas CD44, CXCR4, Tie-2 e Notch-1 nas LT-CTH. Observamos diminuição da expressão da proteína CD44 nos desnutridos. Isolamos as células LT-CTH por cell sorting e avaliamos a expressão gênica de CD44, CXCR4 e NOTCH-1. Verificamos que os desnutridos apresentaram menor expressão de CD44. Em relação ao ciclo celular, verificamos maior quantidade de LT-CTH nas fases G0/G1. Caracterizamos as alterações do tecido ósseo femoral, in vivo. Observamos diminuição da densidade mineral óssea e da densidade medular nos desnutridos. A desnutrição acarretou diminuição da área média das seções transversais, do perímetro do periósteo e do endósteo na cortical do fêmur dos animais. E na região trabecular, verificou-se diminuição da razão entre volume ósseo e volume da amostra e do número de trabéculas, aumento da distância entre as trabéculas e prevalência de trabéculas ósseas em formato cilíndrico. Avaliamos a expressão de colágeno, osteonectina (ON) e osteocalcina (OC) por imuno-histoquímica, e de osteopontina (OPN) por imunofluorescência no fêmur e verificamos diminuição da marcação para OPN, colágeno tipo I, OC e ON nos desnutridos. Evidenciamos, pela técnica do Picrosírius, desorganização na distribuição das fibras colágenas e presença de fibras tipo III nos fêmures dos desnutridos, além de maior número de osteoclastos evidenciados pela reação da fosfatase ácida tartarato resistente. Os osteoblastos da região femoral foram isolados por depleção imunomagnética, imunofenotipados por citometria de fluxo e cultivados em meio de indução osteogênica. Observamos menor positividade para fosfatase alcalina e vermelho de alizarina nas culturas dos osteoblastos dos desnutridos. Avaliamos, por Western Blotting, a expressão de colágeno tipo I, OPN, osterix, Runx2, RANKL e osteoprotegerina (OPG), e, por PCR em tempo real, a expressão de COL1A2, SP7, CXCL12, ANGPT1, SPP1, JAG2 e CDH2 nos osteoblastos isolados. Verificamos que a desnutrição acarretou diminuição da expressão proteica de osterix e OPG e menor expressão gênica de ANGPT1. Avaliamos a proliferação das células LSK (Lin-Sca1+c-Kit+) utilizando ensaio de CFSE (carboxifluoresceína succinimidil ester). Foi realizada cocultura de células LSK e osteoblastos (MC3T3-E1) na presença e ausência de anti-CD44. Após uma semana, verificamos menor proliferação das LSK dos desnutridos. O bloqueio de CD44 das LSK do grupo controle diminuiu a proliferação destas em três gerações. Entretanto, nos desnutridos, esse bloqueio não afetou a proliferação. Concluímos que a DP promoveu alterações no tecido ósseo e nas CTH. Entretanto, não podemos afirmar que as alterações observadas no sistema hemopoético foram decorrentes de alterações exclusivas do nicho endosteal.

Risco de fratura da anca na população idosa institucionalizada no distrito de Portalegre: a influência dos fatores de risco da osteoporose

A osteoporose é uma doença caracterizada pela diminuição da massa óssea e deterioração da microarquitectura do tecido ósseo, levando à fragilidade mecânica e consequente predisposição a fraturas. As fraturas relacionadas com a osteoporose, além de poderem provocar consequências fatais, também constituem um grave problema de saúde pública com elevados reflexos a nível económico e social. Este estudo surgiu com a finalidade de perceber qual o risco de fratura a que a população idosa institucionalizada no distrito de Portalegre está sujeita e de que forma é que os fatores de risco da osteoporose são uma influência. Para chegar a respostas conclusivas neste âmbito, foram implementados dois instrumentos de avaliação a 278 idosos institucionalizados. Através da realização dos questionários, foi possível verificar que existe falta de conhecimento sobre os riscos da osteoporose e a forte relação que esta tem com os hábitos de vida diários. Associado a este problema, enfatiza-se a necessidade de políticas educativas, orientando a população para a necessidade de estilos de vida saudáveis.

Poly(beta-hydroxybutyrate-co-beta-hydroxyvalerate) supports in vitro osteogenesis

Studies have demonstrated that polymeric biomaterials have the potential to support osteoblast growth and development for bone tissue repair. Poly( beta- hydroxybutyrate- co- beta- hydroxyvalerate) ( PHBV), a bioabsorbable, biocompatible polyhydroxy acid polymer, is an excellent candidate that, as yet, has not been extensively investigated for this purpose. As such, we examined the attachment characteristics, self- renewal capacity, and osteogenic potential of osteoblast- like cells ( MC3T3- E1 S14) when cultured on PHBV films compared with tissue culture polystyrene ( TCP). Cells were assayed over 2 weeks and examined for changes in morphology, attachment, number and proliferation status, alkaline phosphatase ( ALP) activity, calcium accumulation, nodule formation, and the expression of osteogenic genes. We found that these spindle- shaped MC3T3- E1 S14 cells made cell - cell and cell - substrate contact. Time- dependent cell attachment was shown to be accelerated on PHBV compared with collagen and laminin, but delayed compared with TCP and fibronectin. Cell number and the expression of ALP, osteopontin, and pro- collagen alpha 1( I) mRNA were comparable for cells grown on PHBV and TCP, with all these markers increasing over time. This demonstrates the ability of PHBV to support osteoblast cell function. However, a lag was observed for cells on PHBV in comparison with those on TCP for proliferation, ALP activity, and cbfa- 1 mRNA expression. In addition, we observed a reduction in total calcium accumulation, nodule formation, and osteocalcin mRNA expression. It is possible that this cellular response is a consequence of the contrasting surface properties of PHBV and TCP. The PHBV substrate used was rougher and more hydrophobic than TCP. Although further substrate analysis is required, we conclude that this polymer is a suitable candidate for the continued development as a biomaterial for bone tissue engineering.

Long-term loading inhibits ERK1/2 phosphorylation and increases FGFR3 expression in MC3T3-E1 osteoblast cells

Bone tissue homeostasis relies upon the ability of cells to detect and interpret extracellular signals that direct changes in tissue architecture. This study utilized a four-point bending model to create both fluid shear and strain forces (loading) during the time-dependent progression of MC3T3-E1 preosteoblasts along the osteogenic lineage. Loading was shown to increase cell number, alkaline phosphatase (ALP) activity, collagen synthesis, and the mRNA expression levels of Runx2, osteocalcin (OC), osteopontin, and cyclo-oxygenase-2. However, mineralization in these cultures was inhibited, despite an increase in calcium accumulation, suggesting that loading may inhibit mineralization in order to increase matrix deposition. Loading also increased fibroblast growth factor receptor-3 (FGFR3) expression coincident with an inhibition of FGFR1, FGFR4, FGF1, and extracellular signal-related kinase (ERK)1/2 phosphorylation. To examine whether these loading-induced changes in cell phenotype and FGFR expression could be attributed to the inhibition of ERK1/2 phosphorylation, cells were grown for 25 days in the presence of the MEK1/2 inhibitor, U0126. Significant increases in the expression of FGFR3, ALP, and OC were observed, as well as the inhibition of FGFR1, FGFR4, and FGF1. However, U0126 also increased matrix mineralization, demonstrating that inhibition of ERK1/2 phosphorylation cannot fully account for the changes observed in response to loading. in conclusion, this study demonstrates that preosteoblasts are mechanoresponsive, and that long-term loading, whilst increasing proliferation and differentiation of preosteoblasts, inhibits matrix mineralization. In addition, the increase in FGFR3 expression suggests that it may have a role in osteoblast differentiation.