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Authors = Christophe Caneparo

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21 pages, 2961 KiB  
Article
Impact of the Use of 2-Phospho-L Ascorbic Acid in the Production of Engineered Stromal Tissue for Regenerative Medicine
by David Brownell, Laurence Carignan, Reza Alavi, Christophe Caneparo, Maxime Labroy, Todd Galbraith, Stéphane Chabaud, François Berthod, Laure Gibot, François Bordeleau and Stéphane Bolduc
Cells 2025, 14(14), 1123; https://doi.org/10.3390/cells14141123 - 21 Jul 2025
Viewed by 520
Abstract
Tissue engineering enables autologous reconstruction of human tissues, addressing limitations in tissue availability and immune compatibility. Several tissue engineering techniques, such as self-assembly, rely on or benefit from extracellular matrix (ECM) secretion by fibroblasts to produce biomimetic scaffolds. Models have been developed for [...] Read more.
Tissue engineering enables autologous reconstruction of human tissues, addressing limitations in tissue availability and immune compatibility. Several tissue engineering techniques, such as self-assembly, rely on or benefit from extracellular matrix (ECM) secretion by fibroblasts to produce biomimetic scaffolds. Models have been developed for use in humans, such as skin and corneas. Ascorbic acid (vitamin C, AA) is essential for collagen biosynthesis. However, AA is chemically unstable in culture, with a half-life of 24 h, requiring freshly prepared AA with each change of medium. This study aims to demonstrate the functional equivalence of 2-phospho-L-ascorbate (2PAA), a stable form of AA, for tissue reconstruction. Dermal, vaginal, and bladder stroma were reconstructed by self-assembly using tissue-specific protocols. The tissues were cultured in a medium supplemented with either freshly prepared or frozen AA, or with 2PAA. Biochemical analyses were performed on the tissues to evaluate cell density and tissue composition, including collagen secretion and deposition. Histology and quantitative polarized light microscopy were used to evaluate tissue architecture, and mechanical evaluation was performed both by tensiometry and atomic force microscopy (AFM) to evaluate its macroscopic and cell-scale mechanical properties. The tissues produced by the three ascorbate conditions had similar collagen deposition, architecture, and mechanical properties in each organ-specific stroma. Mechanical characterization revealed tissue-specific differences, with tensile modulus values ranging from 1–5 MPa and AFM-derived apparent stiffness in the 1–2 kPa range, reflecting the nonlinear and scale-dependent behavior of the engineered stroma. The results demonstrate the possibility of substituting AA with 2PAA for tissue engineering. This protocol could significantly reduce the costs associated with tissue production by reducing preparation time and use of materials. This is a crucial factor for any scale-up activity. Full article
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14 pages, 9014 KiB  
Article
Correction of Significant Urethral Anomalies Using a Tissue-Engineered Human Urethral Substitute: Proof of Concept
by Christophe Caneparo, Elissa Elia, Stéphane Chabaud, François Berthod, Julie Fradette and Stéphane Bolduc
Int. J. Mol. Sci. 2025, 26(5), 1825; https://doi.org/10.3390/ijms26051825 - 20 Feb 2025
Viewed by 1192
Abstract
Urethral reconstruction remains a challenge. Indeed, the use of oral mucosa, the reference biomaterial for urethroplasty, is associated with two main drawbacks: the limited availability of autologous tissues and potential short- and long-term complications, especially for patients with recurrences or severe anomalies. Therefore, [...] Read more.
Urethral reconstruction remains a challenge. Indeed, the use of oral mucosa, the reference biomaterial for urethroplasty, is associated with two main drawbacks: the limited availability of autologous tissues and potential short- and long-term complications, especially for patients with recurrences or severe anomalies. Therefore, the development of alternative approaches, such as urethral tissue engineering, is necessary. A new type of human urethral substitute devoid of exogenous biomaterials has been reconstructed in vitro. It presented sufficient mechanical strength and had histological and functional properties comparable to native tissues. These reconstructed tissues were implanted in vivo to repair hypospadias induced in tacrolimus-immunosuppressed rabbits via a two-stage urethroplasty. In the first stage, the distal part of the native urethra was removed, and a flat graft was implanted, leaving the urethra open proximally. Twelve weeks later, the graft was tubularized to create a neourethra, reproducing the usual clinical scenario. The results obtained for the experimental group were less effective than for the control group, with a success rate of 50% after excluding the animal affected by unwanted events unrelated to urethroplasty, and it is possible that the animal model or surgical technique used was not suitable and should be modified. Nevertheless, half of the urethral substitutes grafted on rabbits showed successful integration. These self-assembled artificial tissues represent promising substitutes for urethroplasty. Full article
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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40 pages, 8196 KiB  
Review
Impact of Endocrine Disruptors on the Genitourinary Tract
by Christophe Caneparo, Laurence Carignan, Elena Lonina, Sarah-Maude Goulet, Felix-Antoine Pellerin, Stéphane Chabaud, François Bordeleau, Stéphane Bolduc and Martin Pelletier
J. Xenobiot. 2024, 14(4), 1849-1888; https://doi.org/10.3390/jox14040099 - 2 Dec 2024
Cited by 3 | Viewed by 3464
Abstract
Over the last decades, the human species has seen an increase in the incidence of pathologies linked to the genitourinary tract. Observations in animals have allowed us to link these increases, at least in part, to changes in the environment and, in particular, [...] Read more.
Over the last decades, the human species has seen an increase in the incidence of pathologies linked to the genitourinary tract. Observations in animals have allowed us to link these increases, at least in part, to changes in the environment and, in particular, to an increasing presence of endocrine disruptors. These can be physical agents, such as light or heat; natural products, such as phytoestrogens; or chemicals produced by humans. Endocrine disruptors may interfere with the signaling pathways mediated by the endocrine system, particularly those linked to sex hormones. These factors and their general effects are presented before focusing on the male and female genitourinary tracts by describing their anatomy, development, and pathologies, including bladder and prostate cancer. Full article
(This article belongs to the Special Issue The Role of Endocrine-Disrupting Chemicals in the Human Health)
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23 pages, 3140 KiB  
Review
Tissue Engineering for Penile Reconstruction
by Elissa Elia, Christophe Caneparo, Catherine McMartin, Stéphane Chabaud and Stéphane Bolduc
Bioengineering 2024, 11(3), 230; https://doi.org/10.3390/bioengineering11030230 - 28 Feb 2024
Cited by 4 | Viewed by 13062
Abstract
The penis is a complex organ with a development cycle from the fetal stage to puberty. In addition, it may suffer from either congenital or acquired anomalies. Penile surgical reconstruction has been the center of interest for many researchers but is still challenging [...] Read more.
The penis is a complex organ with a development cycle from the fetal stage to puberty. In addition, it may suffer from either congenital or acquired anomalies. Penile surgical reconstruction has been the center of interest for many researchers but is still challenging due to the complexity of its anatomy and functionality. In this review, penile anatomy, pathologies, and current treatments are described, including surgical techniques and tissue engineering approaches. The self-assembly technique currently applied is emphasized since it is considered promising for an adequate tissue-engineered penile reconstructed substitute. Full article
(This article belongs to the Special Issue Artificial Organs and Biofabrication of Human Organs)
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17 pages, 1771 KiB  
Article
Evaluation of a Serum-Free Medium for Human Epithelial and Stromal Cell Culture
by Christophe Caneparo, Stéphane Chabaud, Julie Fradette and Stéphane Bolduc
Int. J. Mol. Sci. 2022, 23(17), 10035; https://doi.org/10.3390/ijms231710035 - 2 Sep 2022
Cited by 12 | Viewed by 4331
Abstract
Over the past decade, growing demand from many domains (research, cosmetics, pharmaceutical industries, etc.) has given rise to significant expansion of the number of in vitro cell cultures. Despite the widespread use of fetal bovine serum, many issues remain. Among them, the whole [...] Read more.
Over the past decade, growing demand from many domains (research, cosmetics, pharmaceutical industries, etc.) has given rise to significant expansion of the number of in vitro cell cultures. Despite the widespread use of fetal bovine serum, many issues remain. Among them, the whole constitution of most serums remains unknown and is subject to significant variations. Furthermore, the presence of potential contamination and xenogeny elements is challenging for clinical applications, while limited production is an obstacle to the growing demand. To circumvent these issues, a Serum-Free Medium (SFM) has been developed to culture dermal and vesical fibroblasts and their corresponding epithelial cells, namely, keratinocytes and urothelial cells. To assess the impact of SFM on these cells, proliferation, clonogenic and metabolic assays have been compared over three passages to conditions associated with the use of a classic Fetal Bovine Serum-Containing Medium (FBSCM). The results showed that the SFM enabled fibroblast and epithelial cell proliferation while maintaining a morphology, cell size and metabolism similar to those of FBSCM. SFM has repeatedly been found to be better suited for epithelial cell proliferation and clonogenicity. Fibroblasts and epithelial cells also showed more significant mitochondrial metabolism in the SFM compared to the FBSCM condition. However, the SFM may need further optimization to improve fibroblast proliferation. Full article
(This article belongs to the Special Issue Tissue Engineering and Cell Therapy)
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15 pages, 3232 KiB  
Article
Heat-Inactivation of Fetal and Newborn Sera Did Not Impair the Expansion and Scaffold Engineering Potentials of Fibroblasts
by Félix-Antoine Pellerin, Christophe Caneparo, Ève Pellerin, Stéphane Chabaud, Martin Pelletier and Stéphane Bolduc
Bioengineering 2021, 8(11), 184; https://doi.org/10.3390/bioengineering8110184 - 13 Nov 2021
Cited by 7 | Viewed by 3927
Abstract
Heat inactivation of bovine sera is routinely performed in cell culture laboratories. Nevertheless, it remains debatable whether it is still necessary due to the improvement of the production process of bovine sera. Do the benefits balance the loss of many proteins, such as [...] Read more.
Heat inactivation of bovine sera is routinely performed in cell culture laboratories. Nevertheless, it remains debatable whether it is still necessary due to the improvement of the production process of bovine sera. Do the benefits balance the loss of many proteins, such as hormones and growth factors, that are very useful for cell culture? This is even truer in the case of tissue engineering, the processes of which is often very demanding. This balance is examined here, from nine populations of fibroblasts originating from three different organs, by comparing the capacity of adhesion and proliferation of cells, their metabolism, and the capacity to produce the stroma; their histological appearance, thickness, and mechanical properties were also evaluated. Overall, serum inactivation does not appear to provide a significant benefit. Full article
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26 pages, 2870 KiB  
Review
Genitourinary Tissue Engineering: Reconstruction and Research Models
by Christophe Caneparo, David Brownell, Stéphane Chabaud and Stéphane Bolduc
Bioengineering 2021, 8(7), 99; https://doi.org/10.3390/bioengineering8070099 - 13 Jul 2021
Cited by 20 | Viewed by 5365
Abstract
Tissue engineering is an emerging field of research that initially aimed to produce 3D tissues to bypass the lack of adequate tissues for the repair or replacement of deficient organs. The basis of tissue engineering protocols is to create scaffolds, which can have [...] Read more.
Tissue engineering is an emerging field of research that initially aimed to produce 3D tissues to bypass the lack of adequate tissues for the repair or replacement of deficient organs. The basis of tissue engineering protocols is to create scaffolds, which can have a synthetic or natural origin, seeded or not with cells. At the same time, more and more studies have indicated the low clinic translation rate of research realised using standard cell culture conditions, i.e., cells on plastic surfaces or using animal models that are too different from humans. New models are needed to mimic the 3D organisation of tissue and the cells themselves and the interaction between cells and the extracellular matrix. In this regard, urology and gynaecology fields are of particular interest. The urethra and vagina can be sites suffering from many pathologies without currently adequate treatment options. Due to the specific organisation of the human urethral/bladder and vaginal epithelium, current research models remain poorly representative. In this review, the anatomy, the current pathologies, and the treatments will be described before focusing on producing tissues and research models using tissue engineering. An emphasis is made on the self-assembly approach, which allows tissue production without the need for biomaterials. Full article
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30 pages, 2461 KiB  
Review
Reconstruction of Vascular and Urologic Tubular Grafts by Tissue Engineering
by Christophe Caneparo, Stéphane Chabaud and Stéphane Bolduc
Processes 2021, 9(3), 513; https://doi.org/10.3390/pr9030513 - 12 Mar 2021
Cited by 10 | Viewed by 6655
Abstract
Tissue engineering is one of the most promising scientific breakthroughs of the late 20th century. Its objective is to produce in vitro tissues or organs to repair and replace damaged ones using various techniques, biomaterials, and cells. Tissue engineering emerged to substitute the [...] Read more.
Tissue engineering is one of the most promising scientific breakthroughs of the late 20th century. Its objective is to produce in vitro tissues or organs to repair and replace damaged ones using various techniques, biomaterials, and cells. Tissue engineering emerged to substitute the use of native autologous tissues, whose quantities are sometimes insufficient to correct the most severe pathologies. Indeed, the patient’s health status, regulations, or fibrotic scars at the site of the initial biopsy limit their availability, especially to treat recurrence. This new technology relies on the use of biomaterials to create scaffolds on which the patient’s cells can be seeded. This review focuses on the reconstruction, by tissue engineering, of two types of tissue with tubular structures: vascular and urological grafts. The emphasis is on self-assembly methods which allow the production of tissue/organ substitute without the use of exogenous material, with the patient’s cells producing their own scaffold. These continuously improved techniques, which allow rapid graft integration without immune rejection in the treatment of severely burned patients, give hope that similar results will be observed in the vascular and urological fields. Full article
(This article belongs to the Special Issue Biomaterials and Tissue Engineering)
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40 pages, 2988 KiB  
Review
Innovative Human Three-Dimensional Tissue-Engineered Models as an Alternative to Animal Testing
by Patrick Bédard, Sara Gauvin, Karel Ferland, Christophe Caneparo, Ève Pellerin, Stéphane Chabaud and Stéphane Bolduc
Bioengineering 2020, 7(3), 115; https://doi.org/10.3390/bioengineering7030115 - 17 Sep 2020
Cited by 111 | Viewed by 23623
Abstract
Animal testing has long been used in science to study complex biological phenomena that cannot be investigated using two-dimensional cell cultures in plastic dishes. With time, it appeared that more differences could exist between animal models and even more when translated to human [...] Read more.
Animal testing has long been used in science to study complex biological phenomena that cannot be investigated using two-dimensional cell cultures in plastic dishes. With time, it appeared that more differences could exist between animal models and even more when translated to human patients. Innovative models became essential to develop more accurate knowledge. Tissue engineering provides some of those models, but it mostly relies on the use of prefabricated scaffolds on which cells are seeded. The self-assembly protocol has recently produced organ-specific human-derived three-dimensional models without the need for exogenous material. This strategy will help to achieve the 3R principles. Full article
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