A scalable 3D tissue culture pipeline to enable functional therapeutic screening for pulmonary fibrosis

Cummins, Katherine A; Bitterman, Peter B.; Tschumperlin, Daniel J.; Wood, David K.

doi:10.1063/5.0054967

Cited by 6 publications

(2 citation statements)

References 61 publications

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“…Microspheres generated using collagen enhance the profibrotic genetic markers of the fibroblasts under the action of TGF-β1, causing cellular contraction, matrix remodeling through metalloproteinases, and increased stiffness due to ECM deposition (Figure a) . In another study, fetal lung fibroblasts in a 3D collagen matrix showed increased viability but limited differentiation to myofibroblasts compared to a 2D collagen-coated culture .…”

Section: Three-dimensional Culture Systemsmentioning

confidence: 97%

Advanced 3D In Vitro Lung Fibrosis Models: Contemporary Status, Clinical Uptake, and Prospective Outlooks

Jain,

Shashi Bhushan,

Natarajan

et al. 2024

ACS Biomater. Sci. Eng.

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Fibrosis has been characterized as a global health problem and ranks as one of the primary causes of organ dysfunction. Currently, there is no cure for pulmonary fibrosis, and limited therapeutic options are available due to an inadequate understanding of the disease pathogenesis. The absence of advanced in vitro models replicating dynamic temporal changes observed in the tissue with the progression of the disease is a significant impediment in the development of novel antifibrotic treatments, which has motivated research on tissue-mimetic three-dimensional (3D) models. In this review, we summarize emerging trends in preparing advanced lung models to recapitulate biochemical and biomechanical processes associated with lung fibrogenesis. We begin by describing the importance of in vivo studies and highlighting the often poor correlation between preclinical research and clinical outcomes and the limitations of conventional cell culture in accurately simulating the 3D tissue microenvironment. Rapid advancement in biomaterials, biofabrication, biomicrofluidics, and related bioengineering techniques are enabling the preparation of in vitro models to reproduce the epithelium structure and operate as reliable drug screening strategies for precise prediction. Improving and understanding these model systems is necessary to find the cross-talks between growing cells and the stage at which myofibroblasts differentiate. These advanced models allow us to utilize the knowledge and identify, characterize, and hand pick medicines beneficial to the human community. The challenges of the current approaches, along with the opportunities for further research with potential for translation in this field, are presented toward developing novel treatments for pulmonary fibrosis.

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Section: Three-dimensional Culture Systemsmentioning

confidence: 97%

Advanced 3D In Vitro Lung Fibrosis Models: Contemporary Status, Clinical Uptake, and Prospective Outlooks

Jain,

Shashi Bhushan,

Natarajan

et al. 2024

ACS Biomater. Sci. Eng.

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“…It has been found that the level of α-SMA expression, a contractile marker associated with fibrosis, was directly proportional to gel compliance, as stiff collagen-based materials promote increased expression of α-SMA and integrins (α1, β1), while collagen gels with lower compliance promote reduced expression of those markers 44 . Furthermore, collagen microspheres with encapsulated fibroblasts from either healthy donors or from IPF patients prepared by flow-focused microfluidics and off-chip fibrillization (Figure 2) have been stimulated to adopt a fibrotic profile, with increased production of ECM remodeling proteins (LOXL1, LOXL2, MMP-2), procollagen 1α2, α-SMA, and evident matrix contraction 45 . More recently, scaffolds prepared with either a low (0.4 mg/ml) or high (3 mg/ml) concentration of collagen, to mimic the healthy vs fibrotic lung, have been used to test the effect of mechanical strain on human fetal lung fibroblasts seeded in 3D collagen-I gels.…”

Section: Polymeric-based Scaffoldsmentioning

confidence: 99%

Taking Advantage of 3D Human Cell Culture Platforms to Study Pulmonary Fibrotic Diseases

Piedade

Santos

Ferreira

et al. 2022

Preprint

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Pulmonary fibrosis consists of progressive and irreversible lung tissue stiffening that is typically associated with organ failure. This is a major health problem and a leading cause of death worldwide. The mainstays of current therapy for lung fibrosis rely on lung transplantation in end-stage fibrotic diseases, which has severe limitations, such as the shortage of organ donors and risk of rejection. There is thus an active search for efficient treatments, which can only be achieved with a better understanding of pulmonary fibrosis pathophysiology. Recent advances in 3D tissue engineering led to the development of platforms for drug testing, and that contribute to a better understanding of pulmonary fibrosis pathophysiology. These complex 3D lung platforms recapitulate lung function, structure, and cell and matrix interactions, therefore providing the means for understanding the mechanisms and mediators involved in the fibrotic process. In this perspective, this review discusses the most relevant 3D cell culture platforms to engineer fibrotic lung models as well as their in vitro applications.

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Exogenous interleukin-1 beta stimulation regulates equine tenocyte function and gene expression in three-dimensional culture which can be rescued by pharmacological inhibition of interleukin 1 receptor, but not nuclear factor kappa B, signaling

et al. 2023

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We investigated how Interleukin 1 beta (IL-1β) impacts equine tenocyte function and global gene expression in vitro and determined if these effects could be rescued by pharmacologically inhibiting nuclear factor-κB (NF-KB) or interleukin 1 signalling. Equine superficial digital flexor tenocytes were cultured in three-dimensional (3D) collagen gels and stimulated with IL-1β for two-weeks, with gel contraction and interleukin 6 (IL6) measured throughout and transcriptomic analysis performed at day 14. The impact of three NF-KB inhibitors on gel contraction and IL6 secretion were measured in 3D culture, with NF-KB-P65 nuclear translocation by immunofluorescence and gene expression by qPCR measured in two-dimensional (2D) monolayer culture. In addition, daily 3D gel contraction and transcriptomic analysis was performed on interleukin 1 receptor antagonist-treated 3D gels at day 14. IL-1β increased NF-KB-P65 nuclear translocation in 2D culture and IL6 secretion in 3D culture, but reduced daily tenocyte 3D gel contraction and impacted > 2500 genes at day 14, with enrichment for NF-KB signaling. Administering direct pharmacological inhibitors of NF-KB did reduce NF-KB-P65 nuclear translocation, but had no effect on 3D gel contraction or IL6 secretion in the presence of IL-1β. However, IL1Ra restored 3D gel contraction and partially rescued global gene expression. Tenocyte 3D gel contraction and gene expression is adversely impacted by IL-1β which can only be rescued by blockade of interleukin 1 receptor, but not NF-KB, signalling.

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A scalable 3D tissue culture pipeline to enable functional therapeutic screening for pulmonary fibrosis

Cited by 6 publications

References 61 publications

Advanced 3D In Vitro Lung Fibrosis Models: Contemporary Status, Clinical Uptake, and Prospective Outlooks

Advanced 3D In Vitro Lung Fibrosis Models: Contemporary Status, Clinical Uptake, and Prospective Outlooks

Taking Advantage of 3D Human Cell Culture Platforms to Study Pulmonary Fibrotic Diseases

Exogenous interleukin-1 beta stimulation regulates equine tenocyte function and gene expression in three-dimensional culture which can be rescued by pharmacological inhibition of interleukin 1 receptor, but not nuclear factor kappa B, signaling

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