Uso de la matriz extracelular como material para ingeniería de tejidos

Introducción: La evaluación de injertos vasculares de submucosa de intestino delgado para la regeneración de vasos sanguíneos ha producido una permeabilidad variable (0-100%) que ha sido concurrente con la variabilidad en las técnicas de fabricación. Metodología: Investigamos los efectos de fabricación en permeabilidad y regeneración en un diseño experimental de 22factorial que combino: 1) preservación (P) o remoción (R) de la capa estratum compactum del intestino, y 2) deshidratada (D) o hidratada (H), dentro de cuatro grupos de estudio (PD, RD, PH, RH). Los injertos fueron implantados en las Arterias Carótidas de porcinos (ID 4.5mm, N=4, 7d). Permeabilidad, trombogenicidad, reacción inflamatoria, vascularización, infiltración de fibroblastos, perfil de polarización de macrófagos y fuerza tensil biaxial fueron evaluadas. Resultados: Todos los injertos PD permanecieron permeables (4/4), pero tuvieron escasa vascularización e infiltración de fibroblastos. El grupo RD permaneció permeable (4/4), presentó una extensa vascularización e infiltración de fibroblastos, y el mayor número del fenotipo de macrófagos (M2) asociado a regeneración. El grupo RH presentó menor permeabilidad (3/4), una extensa vascularización e infiltración de fibroblastos, y un perfil dominante de M2. El grupo PH presentó el menor grado de permeabilidad, y a pesar de mayor infiltración celular que PD, exhibió un fenotipo de macrófagos dominante adverso. La elasticidad de los injertos R evolucionó de una manera similar a las Carótidas nativas (particularmente RD, mientras que los injertos P mantuvieron su rigidez inicial. Discusión: Concluimos que los parámetros de fabricación afectan drásticamente los resultados, siendo los injertos RD los que arrojaron mejores resultados.

Effects of alloying elements on the cytotoxic response of titanium alloys

Titanium alloys, alloys, especially beta-type alloys containing beta-stabilizing elements, constitute a highly versatile category of metallic materials that have been under constant development for application in orthopedics and dentistry. This type of alloy generally presents a high mechanical strength-to-weight ratio, excellent corrosion resistance and low elastic modulus. The purpose of this study is to evaluate the cytotoxicity and adhesion of fibroblast cells on titanium alloy substrates containing Nb, Ta, Zr, Cu, Sn and Mo alloying elements. Cells cultured on polystyrene were used as controls. In vitro results with Vero cells demonstrated that the tested materials, except Cu-based alloy, presented high viability in short-term testing. Adhesion of cells cultured on disks showed no differences between the materials and reference except for the Ti-Cu alloy, which showed reduced adhesion attributed to poor metabolic activity. Titanium alloys with the addition of Nb, Ta, Zr, Sn and Mo elements show a promising potential for biomedical applications. (C) 2011 Elsevier B.V. All rights reserved.

Development of a prototype for measuring plantar forces

Studies on the distribution of plantar pressure between the sole of the foot and the ground were developed before the 19th century. Currently, the most often employed plantar pressure measurement systems are Pedar® and FScan®, which have restrictions such as operational difficulty and high cost. In the present study, a device was constructed from two pressure plates capable of measuring plantar forces in discreet areas of the feet at a low cost, using strain-gages attached to sixteen strategic points of the mechanical elements. Sixteen prismatic beams were soldered to each frame, for which the free extremity of each beam represented a specific point of the foot. Two strain gauges were attached to each beam - one near the upper fixed extremity and the other near the lower fixed extremity. Using a Wheatstone bridge electric circuit, the gauges were used to measure the force acting on the extremity of the beam. Precision and accuracy of the prototype was about 10%. In some measurements, accuracy was 2%. The low precision and accuracy were mainly due to the restrictions of the available equipment, which only permitted four measurements at a time. Thus, it was necessary for participants to stand on the plates four separate times, which signified possible changes in the position of the feet on the pressure plates. Despite some limitations, the aim was achieved. The prototype has been used in some studies and represents a contribution to biomechanics, demonstrating the viability of using strain gauges.

Three-dimensional finite element analysis of stress distribution in retention screws of different crown-implant ratios

The retaining screw of the implant-supported dental prosthesis is the weakest point of the crown/implant system. Furthermore, crown height is another important factor that may increase the lever arm. Therefore, the aim of this study was to assess the stress distribution in implant prosthetic screws with different heights of the clinical crown of the prosthesis using the method of three-dimensional finite element analysis. Three models were created with implants (3.75 mm × 10 mm) and crowns (heights of 10, 12.5 and 15 mm). The results were visualised by means of von Mises stress maps that increased the crown heights. The screw structure exhibited higher levels of stresses in the oblique load. The oblique loading resulted in higher stress concentration when compared with the axial loading. It is concluded that the increase of the crown was damaging to the stress distribution on the screw, mainly in oblique loading. © 2013 Taylor & Francis.

Estudos comparativos de três diferentes scaffolds pra crescimento de célula tronco mesenquimal, fibroblastos e queratinócitos

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Zirconia-based dental crown to support a removable partial denture: a three-dimensional finite element analysis using contact elements and micro-CT data

Veneer fracture is the most common complication in zirconia-based restorations. The aim of this study was to evaluate the mechanical behavior of a zirconia-based crown in a lower canine tooth supporting removable partial denture (RPD) prosthesis, varying the bond quality of the veneer/coping interface. Microtomography (μCT) data of an extracted left lower canine were used to build the finite element model (M) varying the core material (gold core - MAu; zirconia core - MZi) and the quality of the veneer/core interface (complete bonded - MZi; incomplete bonded - MZi-NL). The incomplete bonding condition was only applied for zirconia coping by using contact elements (Target/Contact) with 0.3 frictional coefficients. Stress fields were obtained using Ansys Workbench 10.0. The loading condition (L = 1 N) was vertically applied at the base of the RPD prosthesis metallic support towards the dental apex. Maximum principal (σmax) and von Mises equivalent (σvM) stresses were obtained. The σmax (MPa) for the bonded condition was similar between gold and zirconia cores (MAu, 0.42; MZi, 0.40). The incomplete bonded condition (MZi-NL) raised σmax in the veneer up to 800% (3.23 MPa) in contrast to the bonded condition. The peak of σvM increased up to 270% in the MZi-NL. The incomplete bond condition increasing the stress in the veneer/zirconia interface.

MATERIAL MODELING AND ANALYSIS FOR THE DEVELOPMENT OF A REALISTIC BLAST HEADFORM

Blast traumatic brain injury (BTBI) has become an important topic of study because of the increase of such incidents, especially due to the recent growth of improvised explosive devices (IEDs). This thesis discusses a project in which laboratory testing of BTBI was made possible by performing blast loading on experimental models simulating the human head. Three versions of experimental models were prepared – one having a simple geometry and the other two having geometry similar to a human head. For developing the head models, three important parts of the head were considered for material modeling and analysis – the skin, skull and brain. The materials simulating skin, skull and brain went through many testing procedures including dynamic mechanical analysis (DMA). For finding a suitable brain simulant, several materials were tested under low and high frequencies. Step response analysis, rheometry and DMA tests were performed on materials such as water based gels, oil based mixtures and silicone gels cured at different temperatures. The gelatins and silicone gels showed promising results toward their use as brain surrogate materials. Temperature degradation tests were performed on gelatins, indicating the fast degradation of gelatins at room temperature. Silicone gels were much more stable compared to the water based gels. Silicone gels were further processed using a thinner-type additive gel to bring the dynamic modulus values closer to those of human brain matter. The obtained values from DMA were compared to the values for human brain as found in literature. Then a silicone rubber brain mold was prepared to give the brain model accurate geometry. All the components were put together to make the entire head model. A steel mount was prepared to attach the head for testing at the end of the shock tube. Instrumentation was implemented in the head model to obtain effective results for understanding more about the possible mechanisms of BTBI. The final head model was named the Realistic Explosive Dummy Head or the “RED Head.” The RED Head offered potential for realistic experimental testing in blast loading conditions by virtue of its material properties and geometrical accuracy.

Local strain distribution in real three-dimensional alveolar geometries

Mechanical ventilation is not only a life saving treatment but can also cause negative side effects. One of the main complications is inflammation caused by overstretching of the alveolar tissue. Previously, studies investigated either global strains or looked into which states lead to inflammatory reactions in cell cultures. However, the connection between the global deformation, of a tissue strip or the whole organ, and the strains reaching the single cells lining the alveolar walls is unknown and respective studies are still missing. The main reason for this is most likely the complex, sponge-like alveolar geometry, whose three-dimensional details have been unknown until recently. Utilizing synchrotron-based X-ray tomographic microscopy, we were able to generate real and detailed three-dimensional alveolar geometries on which we have performed finite-element simulations. This allowed us to determine, for the first time, a three-dimensional strain state within the alveolar wall. Briefly, precision-cut lung slices, prepared from isolated rat lungs, were scanned and segmented to provide a three-dimensional geometry. This was then discretized using newly developed tetrahedral elements. The main conclusions of this study are that the local strain in the alveolar wall can reach a multiple of the value of the global strain, for our simulations up to four times as high and that thin structures obviously cause hotspots that are especially at risk of overstretching.

Second-Generation Orthopaedic Measurement Device for in-vivo Assessment of Bone Quality

There are two main types of bone in the human body, trabecular and cortical bone. Cortical bone is primarily found on the outer surface of most bones in the body while trabecular bone is found in vertebrae and at the end of long bones (Ross 2007). Osteoporosis is a condition that compromises the structural integrity of trabecular bone, greatly reducing the ability of the bone to absorb energy from falls. The current method for diagnosing osteoporosis and predicting fracture risk is measurement of bone mineral density. Limitations of this method include dependence on the bone density measurement device and dependence on type of test and measurement location (Rubin 2005). Each year there are approximately 250,000 hip fractures in the United States due to osteoporosis (Kleerekoper 2006). Currently, the most common method for repairing a hip fracture is a hip fixation surgery. During surgery, a temporary guide wire is inserted to guide the permanent screw into place and then removed. It is believed that directly measuring this screw pullout force may result in a better assessment of bone quality than current indirect measurement techniques (T. Bowen 2008-2010, pers. comm.). The objective of this project is to design a device that can measure the force required to extract this guide wire. It is believed that this would give the surgeon a direct, quantitative measurement of bone quality at the site of the fixation. A first generation device was designed by a Bucknell Biomedical Engineering Senior Design team during the 2008- 2009 Academic Year. The first step of this project was to examine the device, conduct a thorough design analysis, and brainstorm new concepts. The concept selected uses a translational screw to extract the guide wire. The device was fabricated and underwent validation testing to ensure that the device was functional and met the required engineering specifications. Two tests were conducted, one to test the functionality of the device by testing if the device gave repeatable results, and the other to test the sensitivity of the device to misalignment. Guide wires were extracted from 3 materials, low density polyethylene, ultra high molecular weight polyethylene, and polypropylene and the force of extraction was measured. During testing, it was discovered that the spring in the device did not have a high enough spring constant to reach the high forces necessary for extracting the wires without excessive deflection of the spring. The test procedure was modified slightly so the wires were not fully threaded into the material. The testing results indicate that there is significant variation in the screw pullout force, up to 30% of the average value. This significant variation was attributed to problems in the testing and data collection, and a revised set of tests was proposed to better evaluate the performance of the device. The fabricated device is a fully-functioning prototype and further refinements and testing of the device may lead to a 3rd generation version capable of measuring the screw pullout force during hip fixation surgery.

Population-based design of mandibular fixation plates with bone quality and morphology considerations

In this paper we present a new population-based implant design methodology, which advances the state-of-the-art approaches by combining shape and bone quality information into the design strategy. The method may enhance the mechanical stability of the fixation and reduces the intra-operative in-plane bending which might impede the functionality of the locking mechanism. The computational method is presented for the case of mandibular locking fixation plates, where the mandibular angle and the bone quality at screw locations are taken into account. The method automatically derives the mandibular angle and the bone thickness and intensity values at the path of every screw from a set of computed tomography images. An optimization strategy is then used to optimize the two parameters of plate angle and screw position. The method was applied to two populations of different genders. Results for the new design are presented along with a comparison with a commercially available mandibular locking fixation plate (MODUS(®) TriLock(®) 2.0/2.3/2.5, Medartis AG, Basel, Switzerland). The proposed designs resulted in a statistically significant improvement in the available bone thickness when compared to the standard plate. There is a higher probability that the proposed implants cover areas of thicker cortical bone without compromising the bone mineral density around the screws. The obtained results allowed us to conclude that an angle and screw separation of 129° and 9 mm for females and 121° and 10 mm for males are more suitable designs than the commercially available 120° and 9 mm.

Automated Portal Placement for Arthroscopic Hip Surgery

Placing portal incisions during arthroscopic hip surgery presents challenges for surgeons in terms of anatomic accessibility and patient safety. Based on key anatomic landmarks and portal placement information from recent literature, suggested portal incisions were determined. Guidance in the placement of the three most common portal incision locations (anterior, anterolateral, and posterolateral) for arthroscopic surgery; in addition to visual feedback on tool trajectory to the hip joint is provided in real time by a computer aided system for hip arthroscopy. By simplifying the portal placement process, one of the most challenging aspects of arthroscopic hip surgery, an increased use of this minimally invasive technique could be possible. In addition to portal information, improvements to an existing computer aided system for arthroscopic hip surgery, including a new hip model and redesigned mechanical tracking linkage, were completed.

Use of a 3D perfusion bioreactor with osteoblasts and osteoblast/endothelial cell co-cultures to improve tissue-engineered bone

The delivery of oxygen, nutrients, and the removal of waste are essential for cellular survival. Culture systems for 3D bone tissue engineering have addressed this issue by utilizing perfusion flow bioreactors that stimulate osteogenic activity through the delivery of oxygen and nutrients by low-shear fluid flow. It is also well established that bone responds to mechanical stimulation, but may desensitize under continuous loading. While perfusion flow and mechanical stimulation are used to increase cellular survival in vitro, 3D tissue-engineered constructs face additional limitations upon in vivo implantation. As it requires significant amounts of time for vascular infiltration by the host, implants are subject to an increased risk of necrosis. One solution is to introduce tissue-engineered bone that has been pre-vascularized through the co-culture of osteoblasts and endothelial cells on 3D constructs. It is unclear from previous studies: 1) how 3D bone tissue constructs will respond to partitioned mechanical stimulation, 2) how gene expression compares in 2D and in 3D, 3) how co-cultures will affect osteoblast activity, and 4) how perfusion flow will affect co-cultures of osteoblasts and endothelial cells. We have used an integrated approach to address these questions by utilizing mechanical stimulation, perfusion flow, and a co-culture technique to increase the success of 3D bone tissue engineering. We measured gene expression of several osteogenic and angiogenic genes in both 2D and 3D (static culture and mechanical stimulation), as well as in 3D cultures subjected to perfusion flow, mechanical stimulation and partitioned mechanical stimulation. Finally, we co-cultured osteoblasts and endothelial cells on 3D scaffolds and subjected them to long-term incubation in either static culture or under perfusion flow to determine changes in gene expression as well as histological measures of osteogenic and angiogenic activity. We discovered that 2D and 3D osteoblast cultures react differently to shear stress, and that partitioning mechanical stimulation does not affect gene expression in our model. Furthermore, our results suggest that perfusion flow may rescue 3D tissue-engineered constructs from hypoxic-like conditions by reducing hypoxia-specific gene expression and increasing histological indices of both osteogenic and angiogenic activity. Future research to elucidate the mechanisms behind these results may contribute to a more mature bone-like structure that integrates more quickly into host tissue, increasing the potential of bone tissue engineering.

Exploration of the role of serum factors in maintaining bone mass during hibernation in black bears

Disuse osteoporosis is a condition in which reduced mechanical loading (e.g. bed-rest, immobilization, or paralysis) results in unbalanced bone turnover. The American black bear is a unique, naturally occurring model for the prevention of disuse osteoporosis. Bears remain mostly inactive for up to half a year of hibernation annually, yet they do not lose bone mechanical strength or structural properties throughout hibernation. The long-term goal of this study is to determine the biological mechanism through which bears maintain bone during hibernation. This mechanism could pinpoint new signaling pathway targets for the development of drugs for osteoporosis prevention. In this study, bone specific alkaline phosphatase (BSALP), a marker of osteoblast activity, and tartrate resistant acid phosphatase (TRACP), a marker of osteoclast number, were quantified in the serum of hibernating and active black bears. BSALP and TRACP decreased during hibernation, suggesting a balanced reduction in bone turnover. This decrease in BSALP and TRACP were correlated positively to serum adiponectin and inversely to serum neuropeptide Y, suggesting a possible role of these hormones in suppressing bone turnover during hibernation. Osteocalcin (OCN) and undercarboxylated OCN increased dramatically in the serum of hibernating bears. These increases were inversely correlated with adiponectin, glucose, and serotonin, suggesting that OCN may have a unique role in energy homeostasis during hibernation. Finally, MC3T3-E1 osteoblasts were cultured in the serum from active and hibernating bears, and seasonal cell responses were quantified. Cells cultured in serum from hibernating bears had a reduced caspase-3/7 response, and more living cells, after apoptotic threat. The caspase-3/7 response was positively correlated to serum adiponectin and to gene expression of OCN and Runx2, suggesting that reduced caspase-3/7 activity may be related to the reduced differentiation potential of osteoblasts in hibernation serum, and that adiponectin is a potential effector hormone. In summary, the activities of osteoblasts and osteoclasts are reduced during hibernation in bears. This reduced turnover is due, in part, to hormonal control. Further study of potential effectors adiponectin and neuropeptide Y may provide insight into the biological mechanism through which bears maintain bone throughout hibernation.

An image-based method to automatically propagate bony landmarks: application to computational spine biomechanics.

In attempts to elucidate the underlying mechanisms of spinal injuries and spinal deformities, several experimental and numerical studies have been conducted to understand the biomechanical behavior of the spine. However, numerical biomechanical studies suffer from uncertainties associated with hard- and soft-tissue anatomies. Currently, these parameters are identified manually on each mesh model prior to simulations. The determination of soft connective tissues on finite element meshes can be a tedious procedure, which limits the number of models used in the numerical studies to a few instances. In order to address these limitations, an image-based method for automatic morphing of soft connective tissues has been proposed. Results showed that the proposed method is capable to accurately determine the spatial locations of predetermined bony landmarks. The present method can be used to automatically generate patient-specific models, which may be helpful in designing studies involving a large number of instances and to understand the mechanical behavior of biomechanical structures across a given population.

Image-based biomechanical assessment of vertebral body and intervertebral disc in the human lumbar spine

Life expectancy continuously increases but our society faces age-related conditions. Among musculoskeletal diseases, osteoporosis associated with risk of vertebral fracture and degenerative intervertebral disc (IVD) are painful pathologies responsible for tremendous healthcare costs. Hence, reliable diagnostic tools are necessary to plan a treatment or follow up its efficacy. Yet, radiographic and MRI techniques, respectively clinical standards for evaluation of bone strength and IVD degeneration, are unspecific and not objective. Increasingly used in biomedical engineering, CT-based finite element (FE) models constitute the state-of-art for vertebral strength prediction. However, as non-invasive biomechanical evaluation and personalised FE models of the IVD are not available, rigid boundary conditions (BCs) are applied on the FE models to avoid uncertainties of disc degeneration that might bias the predictions. Moreover, considering the impact of low back pain, the biomechanical status of the IVD is needed as a criterion for early disc degeneration. Thus, the first FE study focuses on two rigid BCs applied on the vertebral bodies during compression test of cadaver vertebral bodies, vertebral sections and PMMA embedding. The second FE study highlights the large influence of the intervertebral disc’s compliance on the vertebral strength, damage distribution and its initiation. The third study introduces a new protocol for normalisation of the IVD stiffness in compression, torsion and bending using MRI-based data to account for its morphology. In the last study, a new criterion (Otsu threshold) for disc degeneration based on quantitative MRI data (axial T2 map) is proposed. The results show that vertebral strength and damage distribution computed with rigid BCs are identical. Yet, large discrepancies in strength and damage localisation were observed when the vertebral bodies were loaded via IVDs. The normalisation protocol attenuated the effect of geometry on the IVD stiffnesses without complete suppression. Finally, the Otsu threshold computed in the posterior part of annulus fibrosus was related to the disc biomechanics and meet objectivity and simplicity required for a clinical application. In conclusion, the stiffness normalisation protocol necessary for consistent IVD comparisons and the relation found between degeneration, mechanical response of the IVD and Otsu threshold lead the way for non-invasive evaluation biomechanical status of the IVD. As the FE prediction of vertebral strength is largely influenced by the IVD conditions, this data could also improve the future FE models of osteoporotic vertebra.