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In the clinical and pharmacological fields, there is a need for the production of tissue-engineered small-diameter blood vessels. We have demonstrated previously that the extracellular matrix (ECM) produced by fibroblasts can be used as a scaffold to support three-dimensional (3D) growth of another cell type. Thus, a resistant tissue-engineered vascular media can be produced when such scaffolds are used to culture smooth muscle cells (SMCs). The present study was designed to develop an anisotropic fibroblastic ECM sheet that could replicate the physiological architecture of blood vessels after being assembled into a small diameter vascular conduit. Anisotropic ECM scaffolds were produced using human dermal fibroblasts, grown on a microfabricated substrate with a specific topography, which led to cell alignment and unidirectional ECM assembly. Following their devitalization, the scaffolds were seeded with SMCs. These cells elongated and migrated in a single direction, following a specific angle relative to the direction of the aligned fibroblastic ECM. Their resultant ECM stained for collagen I and III and elastin, and the cells expressed SMC differentiation markers. Seven days after SMCs seeding, the sheets were rolled around a mandrel to form a tissue-engineered vascular media. The resulting anisotropic ECM and cell alignment induced an increase in the mechanical strength and vascular reactivity in the circumferential direction as compared to unaligned constructs. Copyright © 2016 John Wiley & Sons, Ltd.

Promotion of skin repair for acute or chronic wounds through the use of tissue-engineered products is an active field of research. This study evaluates the effects mediated by tissue-engineered biological dressings containing human in vitro-differentiated adipocytes and adipose-derived stromal cells (ASCs). Re-epithelialization, granulation tissue formation and neovascularization of full-thickness cutaneous wounds were specifically assessed using a murine model featuring a fluorescent epidermis. In comparison with wounds that did not receive an adipocyte-containing biological dressing, treated wounds displayed a slight but significantly faster wound closure based on macroscopic observations over 18 days. Non-invasive imaging of GFP-expressing keratinocytes determined that the kinetics of re-epithelialization were similar for both groups. Treated wounds featured thicker granulation tissues (1.7-fold, P < 0.0001) enriched in collagens (1.3-fold, P < 0.0104). In addition, wound cryosections labeled for detection of CD31-expressing cells indicated a 2.2-fold (P < 0.0002) increased neovascularization for the treated wounds at the time of terminal biopsy. This is in accordance with the secretion of pro-angiogenic factors detected in media conditioned by the dressings. Taken together, these results establish that a new type of engineered substitutes featuring a mixture of adipocytes and ASCs can promote cutaneous healing when applied as temporary dressings, suggesting their potential relevance for chronic wound management studies.

There is an ongoing clinical need for tissue-engineered small-diameter (

There is a lack of an optimal transplant material for small calibre blood vessels. This could be overcome by tissue engineering. The optimal construct is to be derived from autologous cells and present mechanical resistance comparable to the gold standard, autologous vessels such as the internal mammary artery or the saphenous vein. Our laboratory has developed the self-assembly approach to produce tissue sheets that can be rolled into such vessel substitutes. Over the years, many improvements have been made to the technique to facilitate smooth muscle cell culture and to produce vascular media substitutes with higher circumferential mechanical resistance.

The wind industry has grown over the years along with the collective desire to produce clean energy. High wind potential sites are often located in northern regions where harsh climatic conditions arise. The performance of anemometers, which provide essential measurements to wind turbine operation and wind resource assessment, is then jeopardized by ice build-up and snow accumulation. These conditions result in the need for heated instruments able to remain ice-free regardless of weather conditions while minimizing energy consumption. The main objective of this work was to develop an intelligent heating control for a new generation of ice-free anemometers. The controller is based on vision recognition of ice build-up on the cups of the anemometer using a neural network. Heating power is applied according to the icing state and the icing history of the instrument. Primary results have shown that, for a completely ice-free performance, the developed controller uses an average of less than two thirds of the energy consumed by common control systems similar to those used by other ice-free anemometers available on the market.

There is a clinical need for small-diameter vascular substitutes, notably for coronary and peripheral artery bypass procedures since these surgeries are limited by the availability of grafting material. This study reports the characterization of a novel autologous tissue-engineered vascular substitute (TEVS) produced in 10 weeks exclusively from human adipose-derived stromal cells (ASC) self-assembly, and its comparison to an established model made from dermal fibroblasts (DF). Briefly, ASC and DF were cultured with ascorbate to form cell sheets subsequently rolled around a mandrel. These TEVS were further cultured as a maturation period before undergoing mechanical testing, histological analyses and endothelialization. No significant differences were measured in burst pressure, suture strength, failure load, elastic modulus and failure strain according to the cell type used to produce the TEVS. Indeed, ASC- and DF-TEVS both displayed burst pressures well above maximal physiological blood pressure. However, ASC-TEVS were 1.40-fold more compliant than DF-TEVS. The structural matrix, comprising collagens type I and III, fibronectin and elastin, was very similar in all TEVS although histological analysis showed a wavier and less dense collagen matrix in ASC-TEVS. This difference in collagen organization could explain their higher compliance. Finally, human umbilical vein endothelial cells (HUVEC) successfully formed a confluent endothelium on ASC and DF cell sheets, as well as inside ASC-TEVS. Our results demonstrated that ASC are an alternative cell source for the production of TEVS displaying good mechanical properties and appropriate endothelialization.

Tissue engineering appears as a promising option to create new heart valve substitutes able to overcome the serious drawbacks encountered with mechanical substitutes or tissue valves. The objective of this article is to present the construction method of a new entirely biological stentless aortic valve using the self-assembly method and also a first assessment of its behavior in a bioreactor when exposed to a pulsatile flow. A thick tissue was created by stacking several fibroblast sheets produced with the self-assembly technique. Different sets of custom-made templates were designed to confer to the thick tissue a three-dimensional (3D) shape similar to that of a native aortic valve. The construction of the valve was divided in two sequential steps. The first step was the installation of the thick tissue in a flat preshaping template followed by a 4-week maturation period. The second step was the actual cylindrical 3D forming of the valve. The microscopic tissue structure was assessed using histological cross sections stained with Masson's Trichrome and Picrosirius Red. The thick tissue remained uniformly populated with cells throughout the construction steps and the dense extracellular matrix presented corrugated fibers of collagen. This first prototype of tissue-engineered heart valve was installed in a bioreactor to assess its capacity to sustain a light pulsatile flow at a frequency of 0.5 Hz. Under the light pulsed flow, it was observed that the leaflets opened and closed according to the flow variations. This study demonstrates that the self-assembly method is a viable option for the construction of complex 3D shapes, such as heart valves, with an entirely biological material.

Despite a wide panel of tissue-engineering models available for vesical reconstruction, the lack of a differentiated urothelium remains their main common limitation. For the first time to our knowledge, an entirely human vesical equivalent, free of exogenous matrix, has been reconstructed using the self-assembly method. Moreover, we tested the contribution of adipose-derived stromal cells, an easily available source of mesenchymal cells featuring many potential advantages, by reconstructing three types of equivalent, named fibroblast vesical equivalent, adipose-derived stromal cell vesical equivalent and hybrid vesical equivalent--the latter containing both adipose-derived stromal cells and fibroblasts. The new substitutes have been compared and characterized for matrix composition and organization, functionality and mechanical behaviour. Although all three vesical equivalents displayed adequate collagen type I and III expression, only two of them, fibroblast vesical equivalent and hybrid vesical equivalent, sustained the development of a differentiated and functional urothelium. The presence of uroplakins Ib, II and III and the tight junction marker ZO-1 was detected and correlated with impermeability. The mechanical resistance of these tissues was sufficient for use by surgeons. We present here in vitro tissue-engineered vesical equivalents, built without the use of any exogenous matrix, able to sustain mechanical stress and to support the formation of a functional urothelium, i.e. able to display a barrier function similar to that of native tissue.

Nonurological autologous tissues are used for urethral reconstruction to correct urinary tract disorders but are still leading to complications. Other substitutes have been studied on small animal models without great success. For preclinical tests, we selected the porcine model for its similarity to the human urinary tract. Up to now, porcine skin fibroblasts were not able to synthesize enough extracellular matrix under standard conditions to sustain the formation of an adequate tissue for transplantation purposes. Therefore, our goal was to optimize the harvesting site and culture conditions to obtain a thick and easy to handle porcine fibroblast tissue. The oral mucosa was found to be the ideal harvesting site, and a culture temperature of 39°C enabled the formation of a good porcine fibroblast sheet. We successfully superimpose three fibroblast sheets that merged into a thick and resistant tissue where physiological extracellular matrix was produced. Mechanical resistance evaluation by uniaxial traction on the three-layer fibroblast constructs also demonstrated its suitable properties. The production of this porcine connective tissue offers an interesting option in the field of urological tissue engineering. Autologous experiments on a larger animal model are now possible and accessible, allowing the performance of long-term in vivo studies.

Introduction: Due to the complications associated with the use of non-native biomaterials and the lack of local tissues, bioengineered tissues are required for surgical reconstruction of complex urinary tract diseases, including those of the urinary bladder. The selfassembly method of matrix formation using autologous stromal cells obviates the need for exogenous biomaterials. We aimed at creating novel ex-vivo multilayer urinary tissue from a single bladder biopsy.Methods: After isolating urothelial, bladder stromal and smooth muscle cells from bladder biopsies, we produced 2 models of urinary equivalents: (1) the original one with dermal fibroblasts and (2) the new one with bladder stromal cells. Dermal fibroblasts and bladder stromal cells were stimulated to form an extracellular matrix, followed by sequential seeding of smooth muscle cells and urothelial cells. Stratification and cellular differentiation were assessed by histology, immunostaining and electron microscopy. Barrier function was checked with the permeability test. Biomechanical properties were assessed with uniaxinal tensile strength, elastic modulus, and failure strain.Results: Both urinary equivalents could be handled easily and did not contract. Stratified epithelium, intact basement membrane, fused matrix, and prominent muscle layer were detected in both urinary equivalents. Bladder stromal cell-based constructs had terminally differentiated urothelium and more elasticity than dermal fibroblasts-based equivalents. Permeation studies showed that both equivalents were comparable to native tissues.Conclusions: Organ-specific stromal cells produced urinary tissues with more terminally differentiated urothelium and better biomechanical characteristics than non-specific stromal cells. Smooth muscle cells could be incorporated into the selfassembled tissues effectively. This multi-layer tissue can be used as a urethral graft or as a bladder model for disease modelling and pharmacotherapeutic testing.