Quantification of cell contractile behavior based on non-destructive macroscopic measurement of tension forces on bioprinted hydrogel

Pragnere, Sarah; Kholti, Naima El; Gudimard, Leslie; Essayan, Lucie; Marquette, Christophe A.; Petiot, Emma; Pailler-Mattéi, C.

doi:10.1016/j.jmbbm.2022.105365

Cited by 6 publications

(4 citation statements)

References 68 publications

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“…Staining was performed using GROAT's hematoxylin for 30 seconds followed by Ponceau-Fuchsin solution (0.07% (w/v) Ponceau Xylidine, 0.03% (w/v) Fuchsin acid, 0.2% (v/v) glacial acetic acid) phosphomolybdic acid-orange G (4% (w/v) phosphomolybdic acid, 2% (w/v) orange G) and light green (0.2% (w/v) light green, 0.2% (v/v) glacial acetic acid) incubation for 3 minutes. Protocol for immuno histochemical staining of both −SMA and Collagen I proteins were previously described in [28]Sections were finally dehydrated and mounted using Bioumount DPX (Biognost BM500).…”

Section: Histological Stainingmentioning

confidence: 99%

“…Contraction of the bioprinted dermis was followed by recording of macroscopic evolution and expressed as shrinkage factor (%) already described in Pragnere et al (2022) [28] It was evaluated thanks to image analysis using ImageJ software (NIH, Version 1.46). The total area of the construct was measured from a macroscopic construct picture, standardized to the area of the culture well.…”

Section: Bioprinted Construct Contractionmentioning

confidence: 99%

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Deciphering dermal fibroblast behavior in 3D bioprinted dermis contructs

Chastagnier

Essayan

Courtial

et al. 2023

Preprint

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In recent years, numerous strategies have emerged to answer the growing demand for graftable tissues. Tissue engineering and in-vitro production are one of them. Among all the engineered tissues, skin is one of the most advanced. Nevertheless, biofabrication of graftable and fully functional skin substitutes is still far from being reached. Skin reconstruction, particularly dermis, necessitates cultivation and maturation for several weeks (> 3 weeks) to recover the tissue's composition and functions, which prevent its transfer to clinical applications. Thus, several strategies, including 3D bioprinting, have been explored to accelerate these productions. In the present study, based on the successful application of 3D bioprinting achieved by our group for skin reconstruction in 21 days, we propose to detail the biological behaviors and maturation phases occurring in the bioprinted skin construct thanks to a descriptive approach transferred from the bioprocess field. The aim is to comprehensively characterize dermis construct maturation phases (cell proliferation and ECM secretion) to master later the interdependent and consecutive mechanisms involved in in-vitro production. Thus, standardized quantitative techniques were deployed to describe 3D bioprinted dermis proliferation and maturation phases. Then, in a second step, various parameters potentially impacting the dermis reconstruction phases were evaluated to challenge our methodology and reveal the biological behavior described (fibroblast proliferation and migration, cell death, ECM remodeling with MMP secretion). The parameters studied concern the bioprinting practice including various printed geometries, bioink formulations and cellular physiology in relation with their nutritional supplementation with selected medium additives.

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Section: Histological Stainingmentioning

confidence: 99%

Section: Bioprinted Construct Contractionmentioning

confidence: 99%

Deciphering dermal fibroblast behavior in 3D bioprinted dermis contructs

Chastagnier

Essayan

Courtial

et al. 2023

Preprint

Self Cite

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“…While on Earth, a bioprinted tissue is subjected to complex cultivation se-quences, including mostly manual steps inside a sterile biosafety cabinet (i.e., tissue transfer and feeding). [155,156] In space, such types of sequences could strongly limit the deployment of tissue bioprinting strategies. Accordingly, implementing an all-inone integrated and specifically designed bioprinting and cultivation device is strategic and essential.…”

Section: Online Monitoring Bioreactors and Remote Controlmentioning

confidence: 99%

3D Bioprinting in Microgravity: Opportunities, Challenges, and Possible Applications in Space

Ombergen

Chalupa-Gantner

Chansoria

et al. 2023

Adv Healthcare Materials

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Abstract3D bioprinting has developed tremendously in the last couple of years and enables the fabrication of simple, as well as complex, tissue models. The international space agencies have recognized the unique opportunities of these technologies for manufacturing cell and tissue models for basic research in space, in particular for investigating the effects of microgravity and cosmic radiation on different types of human tissues. In addition, bioprinting is capable of producing clinically applicable tissue grafts, and its implementation in space therefore can support the autonomous medical treatment options for astronauts in future long term and far‐distant space missions. The article discusses opportunities but also challenges of operating different types of bioprinters under space conditions, mainly in microgravity. While some process steps, most of which involving the handling of liquids, are challenging under microgravity, this environment can help overcome problems such as cell sedimentation in low viscous bioinks. Hopefully, this publication will motivate more researchers to engage in the topic, with publicly available bioprinting opportunities becoming available at the International Space Station (ISS) in the imminent future.

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“…Recently, in-house protocols were developed to implement and transfer bioprocess principles to the tissue culture and monitoring. Off-line assays have characterised tissue maturation timelines, including cell growth and metabolic activity pro les 19,20 . The current work explores non-destructive monitoring and quali cation tools for controlling the tissue's physicochemical environment, composition, and evolution.…”

Section: Introductionmentioning

confidence: 99%

Exploring multiple bioprocess monitoring modalities for Large-scale 3D Bioprinted Tissue Cultivation

CHASTAGNIER,

Pragnere,

Gimènez

et al. 2024

Preprint

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In tissue engineering (TE) and regenerative medicine (RM), challenges persist in achieving optimal tissue maturation due to uncontrolled physicochemical environments and the necessity for a dynamic nutrient supply. Real-time monitoring tools are crucial to address these challenges effectively. Our study evaluates nondestructive qualification tools for pre-implantation tissue assessment, aiming to enhance their quality assessment capabilities and broaden their biomedical applications. These tools target internal tissue structure, nutritive medium flow paths, and tissue metabolic state. We extend the capabilities of tissue culture monitoring by integrating advanced bioprocess technologies like Raman spectroscopy or in-vivo imaging tools like magnetic resonance imaging (MRI). Through comparative analysis with Computational Fluid Dynamics (CFD) simulations and MRI velocity mapping, we highlight the synergistic relationship between simulation-based and experimental approaches in optimising tissue feeding and oxygenation. MRI emerges as a precious tool for longitudinal tissue development monitoring, surpassing traditional destructive methods. Our findings underscore the importance of dynamic regulation in tissue culture protocols, facilitated by continuous monitoring and adjustment of the physicochemical tissue environment. Based on evidence from industrial cell-culture processes, Raman spectroscopy emerges as a standard tool for monitoring metabolic tissue. These advancements significantly propel RM and TE, paving the way for comprehensive studies and quantitative analyses essential for developing functional engineered tissues across diverse biomedical applications.

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Quantification of cell contractile behavior based on non-destructive macroscopic measurement of tension forces on bioprinted hydrogel

Cited by 6 publications

References 68 publications

Deciphering dermal fibroblast behavior in 3D bioprinted dermis contructs

Deciphering dermal fibroblast behavior in 3D bioprinted dermis contructs

3D Bioprinting in Microgravity: Opportunities, Challenges, and Possible Applications in Space

Exploring multiple bioprocess monitoring modalities for Large-scale 3D Bioprinted Tissue Cultivation

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