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Purpose This study aims to investigate the impact of layer thickness, extrusion temperature, extrusion speed and build plate temperature on the tensile strength, crystallinity achieved during fabrication (herein, in-process crystallinity) and mesostructure of Poly(lactic acid) specimens. Both tensile strength and in-process crystallinity were optimized and verified as the function of processing parameters, and their relationship was thoroughly examined. Design/methodology/approach The four key technological parameters were systematically varied as factors on three levels, using the statistically designed experiment. Surface response methodology was used to optimize tensile strength and crystallinity for the given ranges of input factors. Optimized factor settings were used in a set of confirmation runs, where the result of optimization was experimentally confirmed. Material characterization was performed using differential scanning calorimetry and X-ray diffraction analysis, while the effect of processing parameters on mesostructure was examined by scanning electron microscopy. Findings Layer thickness and its quadratic effect are dominant contributors to tensile strength. Significant interaction between layer thickness and extrusion speed implies that these parameters should always be varied simultaneously within designed experiment to obtain adequate process model. As regards, the in-process crystallinity, extrusion speed is part of two significant interactions with plate temperature and layer thickness, respectively. Quality of mesostructure is vital contributor to tensile strength during FDM process, while the in-process crystallinity exhibited no impact, remaining below the 20 per cent margin regardless of process parameter settings. Originality/value According to available literature, there have been no previously published investigations which studied the effect of process parameters on tensile strength, mesostructure and in-process crystallinity through systematic variation of four critical processing parameters.

International audience; Layer-by-layer (LBL) BioAssembly method was developed to enhance the control of cell distribution within 3D scaffolds for tissue engineering applications. The objective of this study was to evaluate in vivo the development of blood vessels within LBL bioassembled membranes seeded with human primary cells, and to compare it to cellularized massive scaffolds. Poly(lactic) acid (PLA) membranes fabricated by fused deposition modeling were seeded with monocultures of human bone marrow stromal cells or with cocultures of these cells and endothelial progenitor cells. Then, four cellularized membranes were assembled in LBL constructs. Early osteoblastic and endothelial cell differentiation markers, alkaline phosphatase, and von Willebrand's factor, were expressed in all layers of assemblies in homogenous manner. The same kind of LBL assemblies as well as cellularized massive scaffolds was implanted subcutaneously in mice. Human cells were observed in all scaffolds seeded with cells, but not in the inner parts of massive scaffolds. There were significantly more blood vessels observed in LBL bioassemblies seeded with cocultures compared to all other samples. LBL bioassembly of PLA membranes seeded with a coculture of human cells is an efficient method to obtain homogenous cell distribution and blood vessel formation within the entire volume of a 3D composite scaffold.

Purpose This paper aims to propose a vacuum-assisted post-processing method for use in binder jetted technology. The method is based on six key technological parameters and uses standard, commercially available consumables to achieve improvement in tensile strength, as well as the microstructure and porosity of the infiltrated matrix. Design/methodology/approach Six key technological parameters were systematically varied as factors on three levels, using design of experiment, i.e. definitive screening design. Surface response methodology was used to optimize the process and yield optimal tensile strength for the given range of input factors. Thus obtained, the optimized factor settings were used in a set of confirmation runs, where the result of optimization was experimentally confirmed. To confirm improvement in microstructure of the infiltrated matrix, SEM analysis was performed, while the reduction of porosity was analyzed using mercury porosimetry. Findings The obtained results indicate that, compared to its conventional counterpart, the proposed, optimized infiltration method yields improvement in tensile strength which is significant from both the statistical and engineering point of view, while reducing porosity by 3.5 times, using only standard consumables. Scanning electron microscopy examination of fractured specimens’ micrographs also revealed significant morphological differences between the conventional and proposed method of post-processing. This primarily reflects in higher surface area under hardened epoxy infiltrate, which contributes to increased load capacity of specimen cross-section. Research limitations/implications At the present stage of development, the most important limitation of the proposed method is the overall size of models which can be accommodated in standard vacuum impregnation units. Although, in this study, the infiltration method did not prove statistically significant, further investigation is required with models of complex geometry, various sizes and mass arrangements, where infiltration would be more challenging and could possibly result in different findings. Practical implications The most important practical implication of this study is the experimentally verified result of optimization, which showed that tensile strength and matrix microstructure can be significantly improved, using just standard consumables. Social implications Improved strength contributes to reduction of material consumption, which, in a longer run, can be beneficial for environment protection and sustainable development. Originality/value Based on literature review, there have been no previous investigations which studied the tensile strength of infiltrated specimens through design of experiment, which involved specimen preheating temperature, level and duration of vacuum treatment of infiltrate mixture and infiltrated specimens and infiltration method.

Purpose The purpose of this study was to examine the impact of five key build parameters – layer thickness, deposition angle, infill, extrusion speed and extrusion temperature, and their interactions – on the maximum flexural force in specimens which are made of polylactic acid (PLA). Design/methodology/approach Through a previous study on the flexural properties of PLA specimens, a statistically significant effect of layer thickness was indicated, requiring further experimentation to establish the values of quadratic term in the model, as well as to perform optimization. Instead of performing a conventional Central Composite Design, a novel, definitive screening design (DSD) was used as statistical method. DSD allowed the reduction of the number of runs required for optimization while minimizing aliasing. Findings Significance of deposition angle and infill as main effects was established. Moreover, significant two-way interactions between infill/layer thickness and infill/extrusion speed were detected and discussed. The optimization procedure showed that minimum level of deposition angle, maximum levels of extrusion speed and infill and near mid-level of layer thickness yield maximum flexural force. Research limitations/implications In this study, the three levels of infill were 0.1, 0.2 and 0.3, which corresponds to 10, 20 and 30 per cent of infill, respectively. In everyday practice, infill is usually kept within this range since it allows time-efficiency, i.e. significant reduction of build time. Though, unsurprisingly, higher infill is positively correlated with flexural strength, this study provides practical directions for optimal selection of other key parameters when working with low infill values. Social implications Optimal 3D printing with low infill can contribute to lower material waste and pollution, while PLA plastic’s biodegradability remains high on the environment protection agenda. Originality/value According to available literature, no previous studies have investigated the FDM extrusion of PLA material using a combination of low infill, deposition angle, layer thickness, extrusion speed and extrusion temperature.

Purpose – This study aims to investigate the impact of layer thickness, extrusion temperature, extrusion speed and build plate temperature on the tensile strength, crystallinity achieved during fabrication (herein, in-process crystallinity) and mesostructure of Poly(lactic acid) specimens. Both tensile strength and in-process crystallinity were optimized and verified as the function of processing parameters, and their relationship was thoroughly examined. Design/methodology/approach – The four key technological parameters were systematically varied as factors on three levels, using the statistically designed experiment. Surface response methodology was used to optimize tensile strength and crystallinity for the given ranges of input factors. Optimized factor settings were used in a set of confirmation runs, where the result of optimization was experimentally confirmed. Material characterization was performed using differential scanning calorimetry and X-ray diffraction analysis, while the effect of processing parameters on mesostructure was examined by scanning electron microscopy. Findings – Layer thickness and its quadratic effect are dominant contributors to tensile strength. Significant interaction between layer thickness and extrusion speed implies that these parameters should always be varied simultaneously within designed experiment to obtain adequate process model. As regards, the in-process crystallinity, extrusion speed is part of two significant interactions with plate temperature and layer thickness, respectively. Quality of mesostructure is vital contributor to tensile strength during FDM process, while the in-process crystallinity exhibited no impact, remaining below the 20 per cent margin regardless of process parameter settings. Originality/value – According to available literature, there have been no previously published investigations which studied the effect of process parameters on tensile strength, mesostructure and in-process crystallinity through systematic variation of four critical processing parameters.

Purpose – Main purpose is to present methodology which allows efficient hand gesture recognition using low-budget, 5-sensor data glove. To allow widespread use of low-budget data gloves in engineering virtual reality (VR) applications, gesture dictionaries must be enhanced with more ergonomic and symbolically meaningful hand gestures, while providing high gesture recognition rates when used by different seen and unseen users. Design/methodology/approach – The simple boundary-value gesture recognition methodology was replaced by a probabilistic neural network (PNN)-based gesture recognition system able to process simple and complex static gestures. In order to overcome problems inherent to PNN – primarily, slow execution with large training data sets – the proposed gesture recognition system uses clustering ensemble to reduce the training data set without significant deterioration of the quality of training. The reduction of training data set is efficiently performed using three types of clustering algorithms, yielding small number of input vectors that represent the original population very well. Findings – The proposed methodology is capable of providing efficient recognition of simple and complex static gestures and was also successfully tested with gestures of an unseen user, i.e. person who took no part in the training phase. Practical implications – The hand gesture recognition system based on the proposed methodology enables the use of affordable data gloves with a small number of sensors in VR engineering applications which require complex static gestures, including assembly and maintenance simulations. Originality/value – According to literature, there are no similar solutions that allow efficient recognition of simple and complex static hand gestures, based on a 5-sensor data glove.

Purpose – The purpose of this paper is to propose a general model for locating and clamping workpieces of complex geometry with two skewed holes under multiple constraints. Design/methodology/approach – Numerous constraints related to application of the proposed model are discussed as prerequisite to design of fixture solution. Based on theoretical model, a fixture was designed and successfully tested in experimental investigation. Experimental results were also verified using FEM simulations. Findings – This study showed that, opposed to conventional approach, novel solution results in significantly smaller fixture dimensions, while providing greater stability. Insertion of mandrels and supports element sub-assemblies into the workpiece holes significantly increases workpiece stiffness through an increased moment of inertia, while the internal support elements largely diminish the problem of thin wall deformation in the workpiece. Practical implications – The fixture designed in this case was actually us...

International audience; The conventional tissue engineering is based on seeding of macroporous scaffold on its surface ("top-down" approach). The main limitation is poor cell viability in the middle of the scaffold due to poor diffusion of oxygen and nutrients and insufficient vascularization. Layer-by-Layer (LBL) bioassembly is based on "bottom-up" approach, which considers assembly of small cellularized blocks. The aim of this work was to evaluate proliferation and differentiation of human bone marrow stromal cells (HBMSCs) and endothelial progenitor cells (EPCs) in two and three dimensions (2D, 3D) using a LBL assembly of polylactic acid (PLA) scaffolds fabricated by 3D printing. 2D experiments have shown maintain of cell viability on PLA, especially when a co-cuture system was used, as well as adequate morphology of seeded cells. Early osteoblastic and endothelial differentiations were observed and cell proliferation was increased after 7 days of culture. In 3D, cell migration was observed between layers of LBL constructs, as well as an osteoblastic differentiation. These results indicate that LBL assembly of PLA layers could be suitable for BTE, in order to promote homogenous cell distribution inside the scaffold and gene expression specific to the cells implanted in the case of co-culture system.

In the recent years, there has been an increasing interest in the research of biomechanical aspects of biomaterials and human tissues (1-3). Although studies conducted in vivo and in vitro have provided some of the answers in this field, dental and medical research is usually costly, and may be ethically questionable and time-consuming (4,5). Because of this, the use of numerical models and in vitro simulations became a valuable tool for saving time and money associated with laboratory and clinical research (6). Previous studies have reported different techniques for generating three-dimensional (3D) solid models of the teeth (7-9). Nowadays, technological development brings new possibilities for efficient generation of sophisticated 3D solid models. For example, using specialized software, these models can be generated based on computed-tomography (CT) scan data (5,10-12). In addition, the application of finite element analysis (FEA) allows calculation of stress and strain within tooth structure and biomaterials, which can hardly be measured in vivo (13). Cavity design preparation has a great impact on stress values and fracture resistance of a tooth (7,14-16). It is a factor of a paramount importance, especially in cases of restoring maxillary premolars with extensive mesio-occlusal-distal (MOD) cavities (14,17,18). From the biomechanical point of view, different opinions have been reported on the most appropriate restorative procedure in such cases. Kuijs et al have found that ceramic, indirect resin composite and direct resin composite provide comparable fatigue resistance in a cusp replacing restorations (19). These findings were supported by clinical trials performed by van Dijken and Hickel et al (20,21). On the other hand, Soares et al found that MOD cavities restored with resin composite placed with direct technique attained better biomechanical performance than those restored with laboratory processed resin and ceramic restorations (15,16). Another study also confirmed that in comparison with ceramic restorations, resin composite restoration had higher fatigue resistance (17). As opposed to preparation for direct restoration, cavity preparation for indirect restorations requires removal of additional amount of tooth structure (22). The situation is the same with the cavity preparation for the amalgam (23). Since the quantity of the tooth structure removed while doing cavity preparation affects the biomechanical characteristics of the restored tooth, the use of adhesive direct restorations should be recommended for reinforcing the remaining dental structure (2,16). When planning the design of MOD cavity preparation, cavity wall thickness and cusp reduction should be carefully considered. Usually, cusp reduction is recommended when cavity isthmus width is 2/3 of intercuspal width (7,24). Although this promotes more dental tissue reduction (25), it was shown that the reduction of cuspal height by 2.0 mm increases fracture resistance of a premolar when restored with direct resin composite (26,27). On the other hand, cavity wall thickness is not well defined. Macpherson et al found that 2.25 mm wall thickness is critical for restoring fracture resistance of tooth with MOD cavity (28), but another study, which investigated 1.0-3.0 mm wall thicknesses, reported that the thickness of remaining cavity walls was not relevant to fracture resistance (27). The aim of this study was to investigate the effect of cavity design preparation on stress values in remaining tooth structures restored with resin composite. The null hypothesis was that stress values were not significantly influenced by the cavity wall thickness and cusp reduction.

Statement of problem Poly(methyl-methacrylate) (PMMA) represents the most popular current denture material. However, its major drawbacks are insufficient ductility and strength. Purpose The purpose of this study was to improve the mechanical properties of PMMA in denture base application by adding small quantities of nanosilica. Material and method Silica nanoparticles were added to the liquid component of the tested materials. The standard heat polymerizing procedure was followed to obtain 6 PMMA--silicon dioxide (/SiO 2 ) concentrations (0.023%, 0.046%, 0.091%, 0.23%, 0.46%, and 0.91% by volume). Microhardness and fracture toughness of each set of specimens was compared with the unmodified specimens. Furthermore, differential scanning calorimetry and scanning electron microscopy analyses were conducted, and the results obtained were correlated with the results of mechanical properties. Results It was found that the maximum microhardness and fracture toughness values of the materials tested were obtained for the lowest nanosilica content. A nanosilica content of 0.023% resulted in an almost unchanged glass transition temperature (Tg), whereas the maximum amount of nanosilica induced a considerable increase in Tg. A higher Tg indicated the possible existence of a thicker interfacial layer caused by the chain immobility due to the presence of the particles. However, scanning electron microscopy results demonstrated extensive agglomeration at 0.91% nanosilica, which may have prevented the formation of a homogenous reinforced field. At a nanosilica content of 0.023%, no agglomeration was observed, which probably influenced a more homogenous distribution of nanoparticles as well as uniform reinforcing fields. Conclusions Low nanoparticle content yields superior mechanical properties along with the lower cost of nanocomposite synthesis.

Modern market imposes stringent demands regarding the product quality/price ratio, with an ever decreasing time-to-market. Furthermore, products are increasingly manufactured in small batches and high varieties, which requires flexibility not only from manufacturing system but the entire manufacturing process. Such demands require manufacturing systems which are highly automated in the domain of preparation and realization of manufacturing activities, which include fixture design. Owing to present trends towards reduction of lead time and human effort devoted to fixture design, computer aided fixture design has gained a prominent role in computer aided environment. In this paper, a system for computer-aided fixture design is presented and verified. This system comprises methods and techniques for fixture design. The structure of this system is based on modular principle, and uses data base and knowledge base. The system allows fixtures to be designed based on geometric features of workpiece, process planning and machining information. A segment of output results is also shown. Finally, conclusions are presented with directions for future investigation. Key words: Machining fixture, rule-based systems, data base, knowledge base.

Tissue Engineering (TE) is “an interdisciplinary field that applies principles of engineering and the life sciences toward development of biological substitutes that restore, maintain, or improve tissue function”. The First application of TE is to replace damaged tissues by artificial cell-materials products of tissue engineering (TE). Another TE application is to produce 2 or 3 dimensional (2D and 3D) models for biological and pharmacological in vitro studies. These models or tissue replacements can be fabricated using a combination of different interdisciplinary methods of medicine, biology, chemistry, physics, informatics and mechanics, providing specific micro-environment with different cell types, growth factors and matrix.One of the major challenges of tissue engineering is related to limited cell penetration in the inner parts of porous biomaterials. Poor cell viability in the center of engineered tissue is a consequence of limited oxygen and nutrients diffusion due to insufficient vascular network within the entire construct. Layer-by-layer (LBL) BioAssembly is a new approach based on assembly of small cellularized constructs that may lead to homogenous cell distribution and more efficient three dimensional vascularization of large tissue engineering constructs.Our hypothesis is that LBL Bioassembly approach is more suitable for bone regeneration than conventional tissue engineering approach. The primary objective of this thesis was to evaluate the advantages of LBL Bioassembly approach using 3D-printed polymer membranes seeded with human primary cells. We have evaluated the efficiency of vascular network formation in vivo within entire 3D tissue engineering construct using LBL bioassembly approach and comparing it to the conventional approach based on seeding of cells on the surface of massive 3D scaffolds. There was no significant difference in number of formed blood vessels in 3D at the outer parts of constructs implanted subcutaneously in mice 8 weeks post-implantation. But in the inner parts of implants which were not in direct contact with a host tissue, we could observe statistically more blood vessel formation when LBL bioassembly approach was used. This vascular network formation was more important in the case of co-cultures than mono-vultures of HBMSCs.There were several secondary objectives in this work. The first was to fabricate cellularized 3D constructs for bone tissue engineering using poly(lactic) acid (PLA) membranes and human primary cells: human bone marrow stroma cells (HBMSCs) isolated from the bone marrow, and endothelial progenitor cells (EPCs) isolated from the umbilical cord blood. Then, we have compared different Additive manufacturing technologies to fabricate scaffolds: direct 3D printing (3DP) starting from PLA powder dissolved in chloroform and fused deposition modelling (FDM) using a commercial or a custom-made printer with different resolutions.The custom-made printer equipped with 100 μm nozzle allowed the highest level of printing resolution concerning pores shape and size. In the meantime we evaluated different stabilization systems for layer-by-layer assembling of PLA membranes with human primary cells: the use of 3D printed PLA clips provided the most efficient stabilization to stack PLA membranes in 3D. Another advantage of this stabilization system is that it could be implanted together with LBL constructs. Then we investigated the most suitable cell culture system for such constructs and we observed more efficient cell proliferation and differentiation when co-culture system is used, comparing to mono-cultures.LBL bioassembly approach seems to be suitable solution for efficient vascularization within entire large 3D tissue engineering constructs especially when co-cultures of mesenchymal and endothelial cells are used.; L’Ingénierie tissulaire (IT) est un domaine interdisciplinaire qui applique les principes de l'ingénierie et des sciences de la vie au développement de substituts biologiques afin de restaurer, maintenir ou améliorer la fonction tissulaire. Sa première application consiste à remplacer les tissus endommagés par des produits cellulaires artificiels. Une autre application de l’IT est basée sur la production des modèles en 2 et 3 dimensions (2D et 3D) pour des études biologiques et pharmacologiques in vitro. Ces modèles ou remplacements de tissus peuvent être fabriqués en utilisant des différentes méthodes de médecine, biologie, chimie, physique, informatique et mécanique, fournissant un micro-environnement spécifique avec différents types de cellules, facteurs de croissance et matrice. L'un des principaux défis de l'IT la pénétration cellulaire limitée dans les parties internes des biomatériaux poreux. Une faible viabilité cellulaire au centre du produit d'IT est la conséquence de la diffusion limitée d'oxygène et de nutriments du fait d’un réseau vasculaire insuffisant dans l'ensemble de la construction 3D. Le BioAssembage couche-par-couche est une nouvelle approche basée sur l'assemblage de petites constructions cellularisées permettant une distribution cellulaire homogène et une vascularisation plus efficace dans des produits d’IT.Notre hypothèse est que l'approche couche-par-couche est plus adaptée à la régénération osseuse que l'approche conventionnelle de l'IT. L'objectif principal de cette thèse était d'évaluer les avantages de l'approche couche-par-couche en utilisant des membranes de polymères imprimées en 3D et ensemencées avec des cellules primaires humaines. Nous avons évalué l'efficacité de la formation du réseau vasculaire in vivo dans toute la construction 3D en utilisant cette approche et en la comparant à l'approche conventionnelle basée sur l'ensemencement des cellules sur la surface des scaffolds massives. Il n'y avait pas de différence significative dans le nombre de vaisseaux sanguins formés en 3D au niveau des parties externes des constructions implantées en site souscutanée chez des souris. Mais dans les parties internes des implants qui n'étaient pas en contact direct avec un tissu hôte, nous avons pu observer une formation des vaisseaux sanguins statistiquement plus efficace lorsque l'approche du bio-assemblage couche-par-couche a été utilisée. Cette formation de réseau vasculaire était plus importante dans le cas de co-cultures que de mono-cultures.Il y avait plusieurs objectifs secondaires dans ce travail. Le premier était de fabriquer des constructions 3D cellularisées pour l'IT en utilisant des membranes d'acide polylactique (PLA) et des cellules primaires humaines : des cellules de stroma de moelle osseuse humaine (HBMSCs) isolées de la moelle osseuse et des cellules progénitrices endothéliales (EPCs) isolées du sang du cordon ombilical. Ensuite, nous avons comparé différentes technologies de fabrication des scaffolds: impression 3D directe à partir de poudre de PLA et impression par fil fondu en utilisant une imprimante commerciale et une autre fabriquée sur mesure. L'imprimante sur mesure a permis le plus haut niveau de résolution d'impression spécialement adaptée à la forme et la taille des pores. Par ailleurs, nous avons évalué différents systèmes de stabilisation pour l'assemblage couche par couche : l’utilisation de clips en PLA imprimés en 3D a fourni une stabilisation plus efficace pour empiler les membranes PLA couche par couche. Un autre avantage de ce système de stabilisation est qu'il peut être implanté avec des implants. Ensuite, nous avons observé une prolifération et une différenciation cellulaire plus efficaces lorsque le système de co-culture était utilisé, en comparaison avec des mono-cultures.L'approche du bioassemblage couche-par-couche semble être une solution appropriée pour une vascularisation efficace dans des structures 3D entières d'ingénierie tissulaire.

Tissue Engineering (TE) is “an interdisciplinary field that applies principles of engineering and the life sciences toward development of biological substitutes that restore, maintain, or improve tissue function”. The First application of TE is to replace damaged tissues by artificial cell-materials products of tissue engineering (TE). Another TE application is to produce 2 or 3 dimensional (2D and 3D) models for biological and pharmacological in vitro studies. These models or tissue replacements can be fabricated using a combination of different interdisciplinary methods of medicine, biology, chemistry, physics, informatics and mechanics, providing specific micro-environment with different cell types, growth factors and matrix.One of the major challenges of tissue engineering is related to limited cell penetration in the inner parts of porous biomaterials. Poor cell viability in the center of engineered tissue is a consequence of limited oxygen and nutrients diffusion due to insufficient vascular network within the entire construct. Layer-by-layer (LBL) BioAssembly is a new approach based on assembly of small cellularized constructs that may lead to homogenous cell distribution and more efficient three dimensional vascularization of large tissue engineering constructs.Our hypothesis is that LBL Bioassembly approach is more suitable for bone regeneration than conventional tissue engineering approach. The primary objective of this thesis was to evaluate the advantages of LBL Bioassembly approach using 3D-printed polymer membranes seeded with human primary cells. We have evaluated the efficiency of vascular network formation in vivo within entire 3D tissue engineering construct using LBL bioassembly approach and comparing it to the conventional approach based on seeding of cells on the surface of massive 3D scaffolds. There was no significant difference in number of formed blood vessels in 3D at the outer parts of constructs implanted subcutaneously in mice 8 weeks post-implantation. But in the inner parts of implants which were not in direct contact with a host tissue, we could observe statistically more blood vessel formation when LBL bioassembly approach was used. This vascular network formation was more important in the case of co-cultures than mono-vultures of HBMSCs.There were several secondary objectives in this work. The first was to fabricate cellularized 3D constructs for bone tissue engineering using poly(lactic) acid (PLA) membranes and human primary cells: human bone marrow stroma cells (HBMSCs) isolated from the bone marrow, and endothelial progenitor cells (EPCs) isolated from the umbilical cord blood. Then, we have compared different Additive manufacturing technologies to fabricate scaffolds: direct 3D printing (3DP) starting from PLA powder dissolved in chloroform and fused deposition modelling (FDM) using a commercial or a custom-made printer with different resolutions.The custom-made printer equipped with 100 μm nozzle allowed the highest level of printing resolution concerning pores shape and size. In the meantime we evaluated different stabilization systems for layer-by-layer assembling of PLA membranes with human primary cells: the use of 3D printed PLA clips provided the most efficient stabilization to stack PLA membranes in 3D. Another advantage of this stabilization system is that it could be implanted together with LBL constructs. Then we investigated the most suitable cell culture system for such constructs and we observed more efficient cell proliferation and differentiation when co-culture system is used, comparing to mono-cultures.LBL bioassembly approach seems to be suitable solution for efficient vascularization within entire large 3D tissue engineering constructs especially when co-cultures of mesenchymal and endothelial cells are used.; L’Ingénierie tissulaire (IT) est un domaine interdisciplinaire qui applique les principes de l'ingénierie et des sciences de la vie au développement de substituts biologiques afin de restaurer, maintenir ou améliorer la fonction tissulaire. Sa première application consiste à remplacer les tissus endommagés par des produits cellulaires artificiels. Une autre application de l’IT est basée sur la production des modèles en 2 et 3 dimensions (2D et 3D) pour des études biologiques et pharmacologiques in vitro. Ces modèles ou remplacements de tissus peuvent être fabriqués en utilisant des différentes méthodes de médecine, biologie, chimie, physique, informatique et mécanique, fournissant un micro-environnement spécifique avec différents types de cellules, facteurs de croissance et matrice. L'un des principaux défis de l'IT la pénétration cellulaire limitée dans les parties internes des biomatériaux poreux. Une faible viabilité cellulaire au centre du produit d'IT est la conséquence de la diffusion limitée d'oxygène et de nutriments du fait d’un réseau vasculaire insuffisant dans l'ensemble de la construction 3D. Le BioAssembage couche-par-couche est une nouvelle approche basée sur l'assemblage de petites constructions cellularisées permettant une distribution cellulaire homogène et une vascularisation plus efficace dans des produits d’IT.Notre hypothèse est que l'approche couche-par-couche est plus adaptée à la régénération osseuse que l'approche conventionnelle de l'IT. L'objectif principal de cette thèse était d'évaluer les avantages de l'approche couche-par-couche en utilisant des membranes de polymères imprimées en 3D et ensemencées avec des cellules primaires humaines. Nous avons évalué l'efficacité de la formation du réseau vasculaire in vivo dans toute la construction 3D en utilisant cette approche et en la comparant à l'approche conventionnelle basée sur l'ensemencement des cellules sur la surface des scaffolds massives. Il n'y avait pas de différence significative dans le nombre de vaisseaux sanguins formés en 3D au niveau des parties externes des constructions implantées en site souscutanée chez des souris. Mais dans les parties internes des implants qui n'étaient pas en contact direct avec un tissu hôte, nous avons pu observer une formation des vaisseaux sanguins statistiquement plus efficace lorsque l'approche du bio-assemblage couche-par-couche a été utilisée. Cette formation de réseau vasculaire était plus importante dans le cas de co-cultures que de mono-cultures.Il y avait plusieurs objectifs secondaires dans ce travail. Le premier était de fabriquer des constructions 3D cellularisées pour l'IT en utilisant des membranes d'acide polylactique (PLA) et des cellules primaires humaines : des cellules de stroma de moelle osseuse humaine (HBMSCs) isolées de la moelle osseuse et des cellules progénitrices endothéliales (EPCs) isolées du sang du cordon ombilical. Ensuite, nous avons comparé différentes technologies de fabrication des scaffolds: impression 3D directe à partir de poudre de PLA et impression par fil fondu en utilisant une imprimante commerciale et une autre fabriquée sur mesure. L'imprimante sur mesure a permis le plus haut niveau de résolution d'impression spécialement adaptée à la forme et la taille des pores. Par ailleurs, nous avons évalué différents systèmes de stabilisation pour l'assemblage couche par couche : l’utilisation de clips en PLA imprimés en 3D a fourni une stabilisation plus efficace pour empiler les membranes PLA couche par couche. Un autre avantage de ce système de stabilisation est qu'il peut être implanté avec des implants. Ensuite, nous avons observé une prolifération et une différenciation cellulaire plus efficaces lorsque le système de co-culture était utilisé, en comparaison avec des mono-cultures.L'approche du bioassemblage couche-par-couche semble être une solution appropriée pour une vascularisation efficace dans des structures 3D entières d'ingénierie tissulaire.