Intervertebral disc organ culture for the investigation of disc pathology and regeneration – benefits, limitations, and future directions of bioreactors

Pfannkuche, Judith; Guo, Wei; Cui, Shangbin; Ma, Junxuan; Lang, Gernot; Peroglio, Marianna; Richards, R. Geoff; Alini, Mauro; Grad, Sibylle; Li, Zhen

doi:10.1080/03008207.2019.1665652

Cited by 31 publications

(50 citation statements)

References 146 publications

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“…For example, collagen fiber diameter and stiffness can be readily modified based on structural and mechanical changes noted with degeneration, or diseases such as diabetes ( Adams and Roughley, 2006 ; Li et al, 2013 ; Svensson et al, 2018 ). Furthermore, the model can be easily modified to evaluate advanced tissue engineering designs (e.g., angle-ply disc replacements) before conducting costly and time-intensive in vivo studies in large animal models ( Martin et al, 2014 ), or to help track time-dependent changes during bioreactor organ cultures ( Frauchiger et al, 2018 ; Pfannkuche et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

A Robust Multiscale and Multiphasic Structure-Based Modeling Framework for the Intervertebral Disc

Zhou

Lim

O’Connell

2021

Front. Bioeng. Biotechnol.

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A comprehensive understanding of multiscale and multiphasic intervertebral disc mechanics is crucial for designing advanced tissue engineered structures aiming to recapitulate native tissue behavior. The bovine caudal disc is a commonly used human disc analog due to its availability, large disc height and area, and similarities in biochemical and mechanical properties to the human disc. Because of challenges in directly measuring subtissue-level mechanics, such as in situ fiber mechanics, finite element models have been widely employed in spinal biomechanics research. However, many previous models use homogenization theory and describe each model element as a homogenized combination of fibers and the extrafibrillar matrix while ignoring the role of water content or osmotic behavior. Thus, these models are limited in their ability in investigating subtissue-level mechanics and stress-bearing mechanisms through fluid pressure. The objective of this study was to develop and validate a structure-based bovine caudal disc model, and to evaluate multiscale and multiphasic intervertebral disc mechanics under different loading conditions and with degeneration. The structure-based model was developed based on native disc structure, where fibers and matrix in the annulus fibrosus were described as distinct materials occupying separate volumes. Model parameters were directly obtained from experimental studies without calibration. Under the multiscale validation framework, the model was validated across the joint-, tissue-, and subtissue-levels. Our model accurately predicted multiscale disc responses for 15 of 16 cases, emphasizing the accuracy of the model, as well as the effectiveness and robustness of the multiscale structure-based modeling-validation framework. The model also demonstrated the rim as a weak link for disc failure, highlighting the importance of keeping the cartilage endplate intact when evaluating disc failure mechanisms in vitro. Importantly, results from this study elucidated important fluid-based load-bearing mechanisms and fiber-matrix interactions that are important for understanding disease progression and regeneration in intervertebral discs. In conclusion, the methods presented in this study can be used in conjunction with experimental work to simultaneously investigate disc joint-, tissue-, and subtissue-level mechanics with degeneration, disease, and injury.

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Section: Discussionmentioning

confidence: 99%

A Robust Multiscale and Multiphasic Structure-Based Modeling Framework for the Intervertebral Disc

Zhou

Lim

O’Connell

2021

Front. Bioeng. Biotechnol.

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“…Due to the complicated etiology and pathogenesis of disc degeneration, developing a suitable preclinical model is challenging because it should share features that are similar to disc degeneration in human and it should be reliable, reproducible as well as cost and labor efficient. Currently, there are 4 major categories of preclinical IVD degeneration models: (1) genetic model, (2) mechanical loading, (3) direct structural disruption including annulus/nucleus injury, chemical digestion, and endplate injury, and (4) radicular pain [ [17] , [18] , [19] ], which mimic the triggers in various disease phenotypes of IVD degeneration. The current study focused on the simulation of a post-traumatic IVD degeneration phenotype [ 17 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

One strike loading organ culture model to investigate the post-traumatic disc degenerative condition

Zhou

Cui

et al. 2021

Journal of Orthopaedic Translation

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“…As the authors of this chapter are active in the field of musculoskeletal research, examples will be illustrated using connective tissues or cells isolated from bone and joints. These fields often focus on joint-derived tissues that cause clinical problems for healing such as (i) anterior cruciate ligament (ACL) [1][2][3], (ii) cartilage scaffold engineering [4][5][6][7], (iii) intervertebral disc (IVD) regeneration [8][9][10] and (iv) bone regeneration [11][12][13][14][15]. We demonstrate a subset of fluorescent staining and methods to stain cells from bone and jointderived tissues: that is, staining mesenchymal stromal cells, chondrocytes and IVD cells in native tissue or in 3D hydrogel-like scaffolds such as fibrin [16], polyethylene glycol (PEG) [17], and also cells on solid materials such as silk [18].…”

Section: Introduction 11 the Need For 3d Culture Models In Tissue Engineeringmentioning

confidence: 99%

“…In all of these fields, bioreactor models have been developed to better understand the mechanism of mechanical loading on these tissues. However, the combination of novel smart biomaterials can also be studied together or in direct contact with the organ of interest under more realistic conditions [10,[19][20][21]. More generally speaking, as the aim is to develop clinically relevant models considering the integrity of tissue and organ explants, judging cell viability (CV) becomes a major issue that needs to be assessed [22].…”

Section: Introduction 11 the Need For 3d Culture Models In Tissue Engineeringmentioning

confidence: 99%

Mammalian Cell Viability Methods in 3D Scaffolds for Tissue Engineering

Gantenbein¹,

Croft²,

Larraillet³

2020

Fluorescence Methods for Investigation of Living Cells and Microorganisms

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Modern methods have evolved in tissue engineering to evaluate cell viability (CV) in 3D scaffolds and tissues. These involve either the usage of 3D confocal laser microscopy of live or fixed tissues, or separation of cells from the tissue, either live or fixed, and then their analysis by flow cytometry. Generally, working with live cells has the disadvantage that all the scanning needs to be completed immediately at the end of an experiment. Two different approaches can be distinguished: staining intact cell membranes and staining fixed cells. The entire cytoplasm is stained with amine-reactive dyes (ARDs), these use the principle of dead cell exclusion. Here, we list and compare live-cell versus fixed-cells fluorescence-based methods and also show their limitations, especially when working with autofluorescent or cross-linking materials like silk or genipin-reinforced hydrogels. Microscopic techniques have the advantage over flow cytometry-based methods in that these provide the spatial distribution and morphology of the cells. Calcein AM combined with ethidium-homodimer works for most 3D constructs, where no strong fluorescent background is found on the tissue or scaffold. Frequently, however, concentrations and incubation times need to be adjusted for a specific tissue to ensure diffusion of dyes and optimise emittance for detection.

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Intervertebral disc organ culture for the investigation of disc pathology and regeneration – benefits, limitations, and future directions of bioreactors

Cited by 31 publications

References 146 publications

A Robust Multiscale and Multiphasic Structure-Based Modeling Framework for the Intervertebral Disc

A Robust Multiscale and Multiphasic Structure-Based Modeling Framework for the Intervertebral Disc

One strike loading organ culture model to investigate the post-traumatic disc degenerative condition

Mammalian Cell Viability Methods in 3D Scaffolds for Tissue Engineering

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