Orientador: Gustavo Franco Barbosa / Resumo: O presente projeto de pesquisa, tem como objetivo estudar a viabilidade técnica e financeira para implementação de um manipulador robótico colaborativo,para bioimpressão, utilizando seringas para aplicação da biotinta, definindo os parâmetros de impressão, materiais e equipamentos disponíveis. Propõe-se um estudo sobre os desafios da bioimpressão, tanto na área de desenvolvimento de novos equipamentos,processos e materiais, assim como os desafios sobre impressão de tecidos humanos com células vivas. O trabalho foi realizado com base na revisão da literatura, abordando os principais autores e empresas especializadas no tema proposto, de modo a alcançar o objetivo. O presente projeto estudará a viabilidade de implementação das seringas como efetuador em um manipulador robótico. A metodologia consiste em modelo teórico, seleção dos materiais disponíveis, dividido em quatro etapas, onde serão analisados: concepção, modelo 3D, validação da ideia com simulação em ambiente virtual e fabricação do protótipo em um momento futuro. Os métodos aplicados no projeto contam com um enfoque descritivo qualitativo, com o objetivo de estudar as características e equipamentos apresentados para a fabricação de arcabouços (scaffolds) e tecidos com o uso de robô colaborativo. / Abstract: This research project aims to study the technical and financial feasibility of implementing a collaborative robotic manipulator, for bioprinting, using syringes for bioinkapplication, defining the printing parameters, materials and equipment available. A study is proposed on the challenges of bioprinting, both in the area of development of new equipment, processes and materials, as well as the challenges of printing human tissues with living cells. The work was based on the literature review, approaching the main authors and companies specialized in the proposed theme, in order to reach the objective. This project will study the feasibility of implementing syringes as an effector in a robotic manipulator. The methodology consists of theoretical model, selection of available materials, divided into four stages, which will be analyzed: conception, 3D model, idea validation with simulation in virtual environment and prototypefabrication in a future moment. The methods applied in the project have a qualitative descriptive approach, aiming to study the characteristics and equipment presented for the manufacture of scaffolds and fabrics using a collaborative robot. / Mestre

Organ transplantation has made great progress since the first successful kidney transplant in 1953 and now more than one million tissue transplants are performed in the United States every year (www.organdonor.gov/statistics-stories, 2015). However, the hope and success of organ transplants are often overshadowed by their reputation as being notoriously difficult to procure because of donor-recipient matching and availability. In addition, those that are fortunate enough to receive a transplant are burdened with a lifetime of immunosuppressants. The field of regenerative medicine is currently making exceptional progress toward making it possible for a patient to be their own donor. Cells from a patient can be collected, reprogrammed into stem cells, and then differentiated into specific cell types. This technology combined with recent advances in 3D printing provides a unique opportunity. Cells can now be accurately deposited with computerized precision allowing tissue engineering from the inside out (Gill, 2016). However, more work needs to be done as these techniques have yet to be perfected. Bioprinters can cost hundreds of thousands of dollars, and the bioink they consume costs thousands per liter. The resulting cost in development of protocols required for effective tissue printing can thus be cost-prohibitive, limiting the research to labs which can afford this exorbitant cost and in turn slowing the progress made in the eventual creation of patient derived stem cell engineered organs. The objective of my research is to develop a simple and low-cost introductory system for biological additive manufacturing (Otherwise known as 3D bioprinting). To create an easily accessible and cost-effective system several design constraints were implemented. First, the system had to use mechanical components that could be purchased “off-the-shelf” from commonly available retailers. Second, any mechanical components involved had to be easily sterilizable, modifiable, and compatible with open-source software. Third, any customized components had to be fabricated using only 3D printing and basic tools (i.e. saw, screwdriver, and wrench). Fourth, the system and any expendable materials should be financially available to underfunded school labs, in addition to being sterilizable, biocompatible, customizable, and biodegradable. Finally, all hardware and expendables had to be simple enough as to be operated by high school science students.

Controlled drug delivery systems have been utilized to enhance the therapeutic effects of many current drugs by effectively delivering drugs in a time-dependent and repeatable manner. The ability to control the delivery of drugs, whether through sequential, instantaneous, sustained, delayed and/or enhanced release has the potential to provide effective dosing regimens with enhanced therapeutic effects for a plethora of diseases and injuries. For instance, such systems can enhance anti-tumoral responses or, alternatively, promoting tissue regeneration. The current need for organ and tissue replacement, repair and regeneration for patients is continually growing such that supply is not meeting the high demand primarily due to a paucity of donors as well as biocompatibility issues that lead to immune rejection of the transplant. To overcome this problem, scientists working in the field of tissue engineering and regenerative medicine have investigated the use of scaffolds as an alternative to transplantation. These scaffolds are designed to mimic the extracellular matrix (ECM) by providing structural support as well as promoting attachment, proliferation, and differentiation with the goal of yielding functional tissues or organs. Continued advancement and hybrid approaches using different material combinations and printing methodologies will further advance the progress of 3D printing technologies toward developing scaffolds, and other implantable drug delivery devices, capable of being utilized in the clinic. Such advancements will not only make inroads into improving structural integrity of implantable devices but will also provide platforms for controlled drug delivery from such devices. The primary focus of this thesis will be on controlled drug delivery as well as the integration of controlled drug delivery into 3D printed devices aimed at promoting tissue regeneration. We initially assessed the efficacy of a controlled drug delivery system for the treatment of cancer using on-demand, and sustained, release of an anticancer drug, doxorubicin (DOX), for the treatment of melanoma in a murine model. Using a melanoma model, we investigated the antitumor potential of combining ultrasound (US) with poly(lactic-co-glycolic acid) (PLGA) microspheres loaded with DOX. An in vitro release assay demonstrated an ability of US to affect the release kinetics of DOX from DOX-loaded PLGA microspheres by inducing a 12% increase in rate of release where this treatment resulted in synergistic tumor cell (B16-F10 melanoma cells) killing. Melanoma-bearing mice treated intratumorally with DOX (8 µg)-loaded microspheres and subjected to US treatment at the tumor site were shown to significantly extended survival compared to untreated mice or mice subjected to either treatment alone. The synergistic increase in survival of melanoma-challenged mice treated with the combination of US and DOX-loaded microspheres implicates a promising additional tool for combatting an otherwise currently incurable cancer. We then further investigated other novel control drug delivery systems which included a 3D printed device (tube) for the purposes of sequential drug delivery. 3D printed hollow alginate tubes were fabricated through co-axial bioprinting and then injected with PLGA to provide sequential release of distinct fluorescent dyes (model drugs), where fluorescein was initially released from alginate followed by the delayed release (up to 55 h) of rhodamine B in PLGA. With an alginate shell and a PLGA core, the fabricated tubes showed no cytotoxicity when incubated with the human embryonic kidney (HEK293) cell line or bone marrow stromal stem cells (BMSC). Microscale printing through two-photon polymerization (2PP) was then investigated for controlled drug delivery potential. Poly(ethylene glycol) dimethacrylate (PEGDMA) devices were fabricated using a Photonic Professional GT two-photon polymerization system while rhodamine B was homogenously entrapped inside the polymer matrix during photopolymerization. These devices were printed with varying porosity and morphology and using varying printing parameters such as slicing and hatching distance. Overall, tuning the hatching distance, slicing distance, and pore size of the fabricated devices provided control of rhodamine B release due to resulting changes in the motility of the small molecule and its access to structure edges. In general, increased spacing provided higher drug release while smaller spacing resulted in some occlusion, preventing media infiltration and thus resulting in reduced drug release. 2PP was further explored for its ability to tailor topographical cues in addition to controlled drug release. These physical cues, similar to those of the ECM, have been seen to promote differentiation. With 2PP, we explored microscale topographies with nanoscale precision, where different star size topographies were fabricated. It was observed that the smallest star size topographies differentiated human iPSCs towards the endoderm and mesoderm germ layer. Integrating the facility for controlled drug release into 3D printed devices provides a demand for constructs that not only need to fulfill their purpose of temporarily substituting for the missing tissue at the site of injury, but also providing the necessary cues to promote appropriate tissue regeneration. With 3D printing technology, novel drug delivery constructs were fabricated and tested to appraise functionality such as the ability to control drug delivery and the ability to function as a non-toxic medium for cellular attachment, proliferation, and forced differentiation.

Orientador: Gustavo Franco Barbosa / Resumo: Há décadas, a engenharia de tecidos passou a ser considerada em diversas aplicações, um tratamento médico adequado, devido suas excelentes vantagens, além da escassez de órgãos e disponibilidade de tecidos para serem transplantados. Conhecida como regeneração de novos tecidos, esse ramo da engenharia biomédica fundamentada nos conhecimentos de Biologia, Química e Física, torna-se uma grande alternativa quando tratamentos farmacêuticos convencionais não são mais aplicáveis, utilizando-se de três tipos básicos de ferramentas: célula, scaffolds e fator de crescimento. Dessa forma, esse trabalho tem como propósito principal a manufatura de scaffolds, utilizando a tecnologia de impressão 3D a partir de polímeros termoplásticos biodegradáveis e fosfatos de cálcio (em escala micrométrica), com o objetivo de se obter estruturas 3D complexas e porosas que apresentem propriedades mecânicas adequadas (em relação a ossos) e interconectividade estruturada entre os poros. Com os modelos 3D dos scaffolds projetados, e a seleção e preparação dos materiais envolvidos, foram realizados ajustes de parâmetros para o processamento dos scaffolds e posterior fabricação dos mesmos, mediante o uso da tecnologia de manufatura aditiva com bioimpressora de microextrusão que utiliza sistema de distribuição pneumático para extrusão contínua do material. Por fim os scaffolds foram caracterizados por técnica de análise de propriedade mecânica por ensaio de compressão e as amostras avaliadas pelo método de M... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: For decades, tissue engineering has come to be considered in several applications, an adequate medical treatment, due to its excellent advantages, in addition to the scarcity of organs and the availability of tissues to be transplanted. Known as regenerating of new tissues, this branch of biomedical engineering grounded in the knowledge of biology, chemistry and physics, becomes a great alternative when conventional pharmaceutical treatments are no longer applicable, using three basic types of tools: cell, scaffolds and growth factor. Thus, the main purpose of this work is the manufacture of scaffolds, using the technology of 3D printing from biodegradable thermoplastic polymers and calcium phosphates (in micrometric scale), with the objective of obtaining complex and porous 3D structures that present properties mechanical (in relation to bones) and structured interconnectivity between the pores. With the 3D models of the scaffolds designed, and the selection and preparation of the materials involved, adjustments were made to the processing parameters of the scaffolds and their subsequent manufacture, using the technology of additive manufacturing with microextrusion bioprinter that uses pneumatic distribution system for continuous extrusion of the material. Finally, the scaffolds were characterized by technique of mechanical property analysis by compression test and the samples evaluated by Scanning Electron Microscopy (SEM) method. / Mestre

The goal of nerve tissue engineering is to promote and guide axon growth across a site of nerve injury without misdirection. Bioengineered tissue scaffolds have been shown to be promising for the regeneration of damaged peripheral nerves. Schwann cells play a pivotal role following nerve injury by forming aligned “bands of Büngner” that promote and guide axon regeneration into the distal nerve segment. The incorporation of living Schwann cells into various hydrogels has therefore been urged during the fabrication of tissue engineered nerve scaffolds. The aim of this research is to characterize biomaterials suitable for 3D bioplotting of nerve repair scaffolds. Here a novel technique of scaffold fabrication has been optimized to print alginate-based three-dimensional tissue scaffolds containing hyaluronic acid and living Schwann cells. Alginate/hyaluronic acid scaffolds were successfully fabricated with good printability and cell viability. Addition of the polycation polyethyleneimine (PEI) during the fabrication process stabilized the structure of alginate through the formation of a polyelectrolyte complex and had a significant influence on the degree of swelling, degradation rate, mechanical property, and release kinetics of incorporated protein within the scaffolds. A preliminary in vivo study showed the feasibility of implanting 3D printed alginate/hyaluronic acid scaffolds as nerve conduits in Sprague-Dawley (SD) rats with resected sciatic nerves. However alginate/hyaluronic acid scaffolds were found to be unsuitable for axonal regeneration. Further in vitro culture of Schwann cells was performed in collagen type-I, fibrin, fibrin/hyaluronic acid, and their combination with alginate. It was found that Schwann cells had more favorable cell morphology in fibrin/hyaluronic acid or collagen without alginate. Schwann cell proliferation and alignment were better in fibrin/hyaluronic acid. Therefore fibrin/hyaluronic acid is more ideal than most other hydrogel formulations for use in the bioprinting of nerve repair tissue engineering scaffolds, which incorporate cellular elements. As Schwann cells also align along the long axis of the printed fibrin/hyaluronic acid strands, 3D bioprinting of multiple layers of crosslinked fibrin strands can be used to fabricate a nerve conduit mimicking the bands of Büngner.

So-called, “Dynamic biomaterials” comprised of stimuli-responsive hydrogels are useful in a wide variety of biomedical applications including tissue engineering, drug delivery, and biomedical implants. More than 150,000 peer-reviewed articles (as of 2016) have been published on these materials, and more specifically, over 100,000 of these are on the most widely studied, poly(N-isopropylacrylamide). This thermoresponsive polymer in a crosslinked hydrogel network undergoes a large volume phase transition (𝑉/𝑉0 ~ 10 − 100) within a small temperature range (𝑇 ~ 1 − 3𝐾) making it particularly useful for tissue engineering applications because of the ability to control the topographical configuration of cells into tissue modules which can be applied in multiple layers to form three dimensional constructs. Nevertheless, applications with poly(N-isopropylacrylamide) hydrogels are hindered by two key obstacles: 1. there is presently no quantitative prediction of mechanical properties over the volume phase transition and 2. the mechanisms of cell attachment and detachment remain controversial and unclear. Current polymer-solution theory, first postulated by Paul Flory and Maurice Huggins in 1942, successfully predicts hydrogel swelling for non-stimuli-responsive polymers based on an empirically derived interaction parameter. However, for stimuli-responsive polymer hydrogels, this theoretical framework fails to quantitatively predict swelling and mechanical properties of the polymer. Currently, only qualitative agreement with experiment has been shown. Cell-cell and cell-matrix interactions are mediated through proteins collectively known as cell adhesion molecules. For cell-matrix interactions, these are generally the transmembrane protein, integrin, and the serum protein, fibronectin. It is widely accepted that nearly all molecular mechanisms of cell-matrix interactions are dependent on recognition of the peptide sequence Arg-Gly-Asp. However, much less is known about mechanical mechanisms involved in cell-cell and cell-matrix interactions. Obstacles to the advancement of these applications are 1) unclear mechanisms of cell release and 2) extended exposure of cells to hypothermic conditions. The author, in collaboration with others, has published work demonstrating reduced cell exposure to hypothermic conditions during tissue module release and elucidated a mechanism of tissue module release: mechanical strain. The central hypothesis of work in this proposal is that tissue module release occurs due to a mechanical strain-rate coinciding with critical force needed overcome the dynamic bond strength of cell adhesion molecules. Advances in this area could improve biomaterial design and accelerate the field of regenerative medicine by reducing or eliminating the need for allograft transplants. This dissertation project, then, seeks to address these two obstacles through biophysical characterization methods and analysis including: atomic force microscopy, scanning electron microscopy, laser-scanning confocal micrscopy, phase-contrast microscopy, and mass-balance analysis. It is hypothesized that, (1) mechanical properties of PNIPAAm hydrogels are quantitatively predicted based on crosslinker ratio in the water-rich phase, (2) release of cells from micropatterned PNIPAAm hydrogels occurs when the lateral strain in the surface exceeds ϵ > 0.25, and (3) the molecular mechanism of rapid cell release from micro-patterned PNIPAAm hydrogels is mediated by the transmembrance protein integrin and its extracellular matrix receptor, fibronectin. Results from these studies could be useful for improving the design of biomaterials based on PNIPAAm hydrogels for applications in tissue engineering.

La bioimpression assistée par laser (LAB) permet de positionner des gouttesde cellules avec une précision micrométrique. Il est ainsi possible de donner uneorganisation initiale aux cellules au sein d’une structure tissulaire 3D. Notre objectif estd’utiliser le LAB pour reproduire l’histo-architecture du stroma cornéen. Le stroma cornéenest un assemblage transparent de lamelles d’une épaisseur totale de 500 μm. Au sein dechaque lamelle, les fibres de collagène ont une même direction, un même diamètre et sontrégulièrement espacées grâce à la présence de protéoglycanes spécifiques du stromacornéen. Pour reproduire cette organisation, nous avons fait l’hypothèse qu’en alignant desfibroblastes du stroma sur un hydrogel de collagène à l’aide du LAB, il serait possibled’aligner les fibres de collagène dans la même direction. Du fait que les cellules impriméessont vivantes et dynamiques, le motif cellulaire initialement imprimé est soumis à desprocessus d’auto-organisation. Il a donc fallu déterminer les paramètres, à la foisd’impression et de culture, permettant d’obtenir de façon reproductible des alignements decellules stables dans le temps. Grâce à la microscopie à génération de secondeharmonique, le remaniement des fibres de collagène par les fibroblastes cornéens a pu êtreobservé. La direction des fibres de collagène correspond à celle de l’alignement cellulaire.En imprimant les fibroblastes de cornée sur des couches successives de collagène, noussommes parvenus à reproduire les variations de direction des fibres de collagène d’unelamelle à l’autre qui sont observées dans le stroma cornéen natif. / Laser-Assisted Bioprinting allows positioning of cell droplets with amicrometric precision. It is thus possible to give an initial organization to the cells within a3D tissue structure. Our objective is to use LAB to reproduce the corneal stroma histoarchitecture.The corneal stroma is a transparent assembly of lamellae with a totalthickness of 500 μm. Within each lamella, collagen fibers have the same direction, thesame diameter, and a regular spacing thanks to the presence of proteoglycans which arespecific from the corneal stroma. To reproduce this organization, we make the hypothesisthat through corneal fibroblasts alignment, using LAB, on a collagen hydrogel, it would bepossible to align collagen fibers in the same direction. Because printed cells are alive anddynamic, the cell pattern initially printed is subjected to self-organization processes. It isthus necessary to determine the printing and culture parameters that promote reproducibleand stable cell alignments. By using second harmonic generation microscopy, collagenfiber reorganization by corneal fibroblasts has been observed. Collagen fiber direction ismatching with cell alignment. Corneal fibroblasts have been printed on successive collagenlayers; it allows reproducing the variations in collagen fiber direction from one lamella toanother that are observed in the native corneal stroma.

Ce mémoire est consacré à l’étude expérimentale de deux procédés de bioimpression assistée par laser femtoseconde, fonctionnant à 1030 nm. En effet, les lasers femtosecondes constituent un choix intéressant pour la bioimpression: la versatilité des matériaux qui peuvent être déposés et la zone affectée thermiquement négligeable sont des atouts pour l’impression de structures biologiques complexes, sans compromettre la viabilité et la fonctionnalité des matériaux biologiques transférés. Tout d’abord, la bioimpression assistée par laser femtoseconde avec couche absorbante métallique a été étudiée sur une bioimprimante adaptée au transfert de cellules (MODULAB®). Une étude expérimentale a été menée par observation du jet induit par laser grâce à un système d’imagerie résolue en temps (TRI) et par impression sur receveur (puits de culture). La rhéologie de la bioencre, certains paramètres laser, ainsi que la position de focalisation laser ont été variés lors des expériences. Des tests de viabilité cellulaire après l’impression ont permis d’identifier une énergie optimale de 3 μJ. L’étude de la variation de la position de focalisation a permis de prédire la plage de tolérance de la position de focalisation du laser : pour une énergie de 3,5 μJ et une ON équivalente de 0,125, la tolérance maximale dans la direction « z » était de 60 μm pour pouvoir imprimer.Dans un second temps, la bioimpression assistée par laser femtoseconde sans couche absorbante a été étudiée sur un montage expérimental comprenant un réservoir de bioencre via des paramètres opératoires clés (position de focalisation, ouverture numérique de l’objectif de focalisation, diamètre de goutte imprimé, la hauteur du jet de l’impression par TRI, la distance de transfert limite pour imprimer). L’impression était reproductible pour une distance d’impression de 75 % hmax à 100 % hmax, hmax étant la hauteur maximale du jet d’impression pour une condition expérimentale. L’utilisation du réservoir de bioencre a permis de trouver une position de focalisation z tolérante: Δz a été calculée (Zernike et l’aberration sphérique) et mesurée. Expérimentalement, Δz valait de 0 à 60 μm selon la bioencre et l’ON. Elle était maximale à l’ON 0,4. Cette tolérance est grande devant la profondeur de champs dans l’air (4 μm à l’ON 0,4) mais faible au regard de la tolérance sur la position du receveur qui peut subir une variation de 25% hmax, d’après la plage de reproductibilité. / This manuscript deals with the experimental study of two bioprinting processes assisted by femtosecond laser at a wavelength of 1030 nm. Indeed, femtosecond lasers are an interesting choice for bioprinting: the high versatility of materials which can be deposited and the negligible heat affected zone are advantages to print complex biological structures without compromising viability and functionality of the transferred biological materials. Firstly, femtosecond laser assisted bioprinting with a metallic absorbing layer was studied on a bioprinter adapted for cell printing (MODULAB®). An experimental study was conducted, observing the laser induced jet of liquid with a time-resolved imaging system (TRI) and printing on receiver substrates (cell culture well plate). The bioink rheology, some laser parameters, and the laser focus position were changed during the experiments. Cell viability assays after the printing enabled to identify an optimal energy of 3 μJ. The study of the laser focus position variation allowed predicting the tolerance range of the laser focus position: for 3.5 μJ and an equivalent numerical aperture (NA) of 0.125, the maximum tolerance in the “z” direction was of 60 μm in order to print. Secondly, femtosecond laser assisted bioprinting without an absorbing layer was studied on an experimental set-up comprising a reservoir of bioink. Some key operating parameters were studied (focalization position, NA of the focalization objective, printed drop diameter, printing jet height by TRI, maximum transfer distance for printing). The printing was reproducible for a printing distance from 75 % hmax to 100 % hmax, with hmax corresponding to the maximum printing jet height for a given experimental condition. Using the reservoir of bioink enabled to find a tolerant focalization position z: Δz was calculated (Zernike polynomial and the spherical aberration) and measured. Experimentally, Δz ranged from 0 to 60 μm depending on the bioink and the NA. It was maximal at NA 0.4. This tolerance is high compared to the depth of field in the air (4 μm at NA 0.4) but low compared to the tolerance of the receiver substrate position which can vary to 25 % hmax according to the reproducibility range.

Congenital heart defects are the most common category of birth defects, mostly affecting the blood vessels, walls, or valves of the heart. For example, pulmonary atresia occurs when the connection between the right ventricle to the main pulmonary artery is not fully formed. A heart defect such as pulmonary atresia may need surgery to close up any malformations in walls and blood vessels, and unfortunately, because the patients are infants, they will need to undergo several surgeries in their lifetime to accommodate a heart patch that will fit the size of their hearts at each stage of their life. A better solution would be to create a biomimetic vascular patch that could be anatomically accepted by the patient’s body as its own, allowing it to grow with the patient without the residue of scar tissue. Instead of propagating scar tissue in the area, it would propagate healthy cells that would integrate into the surrounding tissue. For this to become a reality, one strategy for a biomimetic vascular patch would be to build it like a blood vessel in layers, beginning with the tunica adventitia. The goal of this thesis is to engineer and design the foundation for a biomimetic vascular patch with a crimped, collagen-integrated scaffold, focusing on optimizing the mechanical properties of the hybrid structure. The crimped structure, using sine waves generated from Python code and fabricated with bioprinting technology, mimics the natural formation of collagen fibers in native blood vessels. Additionally, testing the scaffolds on the Instron allows for characterization of the mechanical behaviors of an optimal and repeatable foundation for a tissue-engineered tunica adventitia. / 2023-01-22T00:00:00Z

Meniscal tears are the most common intra-articular injury of the knee joint. Due to the avascular zone with limited blood supply, treatment of the injury is a complex process. Today, research on the development of efficient treatments and meniscal replacements is of increasing interest. However, there are few alternatives of meniscal replacements available on the market and research has shown uncertain results in their ability to restore the natural biomechanics of the knee joint or prevent development of osteoarthritis. Furthermore there is no comparable method to evaluate tensile stresses caused by axial compressional load on a whole meniscus replacement. Therefore the possibility of knitted casing to contain, fixate and mechanically stabilize a cell seeded bioprinted gel and develop a methodology to characterize its compressional behaviour was analysed. By interlock knitting with segments of partial knit a 3D crescent-shaped biodegradable casing was produced mimicking the dimension of the medial meniscus. In the casing design, an Artelon® Flexband™ was incorporated functioning both as reinforcement at the peripheral rim and as fixation method. Moreover radial threads were added to the casing design by inclusion of weft inlays in the knitting pattern. In the non-destructive characterization of the compressional behaviour of the prototype, axial compressional forces of 10.82 N and 29.77 N were achieved. However the forces achieved were significantly lower if compared to the high force that is applied to the menisci in the knee joint. Furthermore a high influence of the coefficient of friction of the casing in the axial compressional force was concluded. Nevertheless refinements of the methodology are required to perform evaluation with comparable and reliable results.