Establishing principles of macromolecular crowding for in vitro fibrosis research of the vocal fold lamina propria

Graupp, Matthias; Gruber, Hans-Jürgen; Weiss, Gregor; Kiesler, Karl; Bachna-Rotter, Sophie; Friedrich, Gerhard; Gugatschka, Markus

doi:10.1002/lary.25103

Cited by 17 publications

(15 citation statements)

References 20 publications

(31 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since methods to accelerate matrix production are urgently needed in the fields of tissue engineering and regenerative medicine, our goal here was to shed light on the underpinning mechanisms by which Ficoll exposure upregulates ECM assembly, as previously reported. [11][12][13][14][15][16][17][21][22][23]72,73 Rather than just accelerating pro-collagen cleavage and supramolecular assembly, previously thought to be the main drivers, 2,3 we show here that Ficoll also significantly enhances fibronectin assembly ( Fig. 1-3) and that the rate of fibronectin fibrillogenesis regulates the collagen I assembly process.…”

Section: Discussionmentioning

confidence: 45%

“…18% v/v which replicates the crowding level in blood plasma 2,12,20 and has proven effective in enhancing matrix assembly. [11][12][13][14][15][16][17][21][22][23] Even though negatively charged crowding molecules like carrageenan have a stronger enhancing effect on matrix assembly, they have an unequal impact on different matrix components (e.g. they have a much stronger enhancing effect on collagen I than fibronectin) and therefore result in a change in the composition of the ECM.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Fibrillar fibronectin plays a key role as nucleator of collagen I polymerization during macromolecular crowding-enhanced matrix assembly

2019

View full text Add to dashboard Cite

PAPER Viola Vogel et al. Fibrillar fi bronectin plays a key role as nucleator of collagen I polymerization during macromolecular crowding-enhanced matrix assembly Volume Macromolecular crowding is used by tissue engineers to accelerate extracellular matrix assembly in vitro, however, most mechanistic studies focus on the impact of crowding on collagen fiber assembly and largely ignore the highly abundant provisional matrix protein fibronectin. We show that the accelerated collagen I assembly as induced by the neutral crowding molecule Ficoll is regulated by cell access to fibronectin. Ficoll treatment leads to significant increases in the amount of surface adherent fibronectin, which can readily be harvested by cells to speed up fibrillogenesis. FRET studies reveal that Ficoll crowding also upregulates the total amount of fibronectin fibers in a low-tension state through upregulating fibronectin assembly. Since un-stretched fibronectin fibers have more collagen binding sites to nucleate the onset of collagen fibrillogenesis, our data suggest that the Ficoll-induced upregulation of low-tension fibronectin fibers contributes to enhanced collagen assembly in crowded conditions. In contrast, chemical cross-linking of fibronectin to the glass substrate prior to cell seeding prevents early force mediated fibronectin harvesting from the substrate and suppresses upregulation of collagen I assembly in the presence of Ficoll, even though the crowded environment is known to drive enzymatic cleavage of procollagen and collagen fiber formation. To show that our findings can be exploited for tissue engineering applications, we demonstrate that the addition of supplemental fibronectin in the form of an adsorbed coating markedly improves the speed of tissue formation under crowding conditions. † Electronic supplementary information (ESI) available. See

show abstract

Section: Discussionmentioning

confidence: 45%

Section: Introductionmentioning

confidence: 99%

Fibrillar fibronectin plays a key role as nucleator of collagen I polymerization during macromolecular crowding-enhanced matrix assembly

2019

View full text Add to dashboard Cite

show abstract

“…Therefore, most studies have been investigating antifibrotic drugs in different animals or in vitro. In a recent paper, we described the successful establishment of an in vitro model of VF fibrogenesis based on the principles of MMC [ 9 ]. The current project employed human VFF and studied the effects of antifibrotic agents.…”

Section: Discussionmentioning

confidence: 99%

“…We pursued a pilot study using rats’ VFF to establish a fibrogenesis model based on the principles of MMC [ 9 ]. The next step was to transfer these findings into the human setting by employing human VFF and to test different antifibrotic compounds.…”

Section: Introductionmentioning

confidence: 99%

Towards an in vitro fibrogenesis model of human vocal fold scarring

Graupp

Rinner

Frisch

et al. 2018

Eur Arch Otorhinolaryngol

View full text Add to dashboard Cite

BackgroundVocal fold (VF) scarring remains a therapeutic dilemma and challenge in modern laryngology. To facilitate corresponding research, we aimed to establish an in vitro fibrogenesis model employing human VF fibroblasts (hVFF) and the principles of macromolecular crowding (MMC).MethodsFibrogenesis was promoted by addition of transforming growth factor-β1 to standard medium and medium containing inert macromolecules (MMC). Hepatocyte growth factor (HGF) and Botox type A were tested for their antifibrotic properties in various doses. Experiments were analyzed with respect to the biosynthesis of collagen, fibronectin, and α-smooth muscle actin using immunofluorescence, silver stain and western blot.ResultsMMC led to favourable enhanced deposition of collagen and other extracellular matrix components, reflecting fibrotic conditions. Low doses of HGF were able to dampen profibrotic effects. This could not be observed for higher HGF concentrations. Botox type A did not show any effects.ConclusionBased on the principles of MMC we could successfully establish a laryngeal fibrogenesis model employing hVFF. Our finding of dose-dependent HGF effects is important before going into clinical trials in humans and has never been shown before. Our model provides a novel option to screen various potential antifibrotic compounds under standardized conditions in a short time.Electronic supplementary materialThe online version of this article (10.1007/s00405-018-4922-7) contains supplementary material, which is available to authorized users.

show abstract

“…To facilitate research on this field, Graupp et al described an in vitro fibrogenesis model using human vocal fold fibroblasts (hVFF) and the principles of macromolecular crowding (MMC) [20]. MMC promotes fibrogenesis after the addition of transforming growth factor-beta 1 (TGF-β1) to a medium containing inert macromolecules.…”

Section: Introductionmentioning

confidence: 99%

Paracrine Effects Of Adipose-Derived Stromal/Stem Cells And Stromal Vascular Fraction In An In Vitro Fibrogenesis Model Of Human Vocal Fold Scarring

Velier

Mattei

SIMONCINI

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Background: Vocal folds (VF) scarring leads to severe dysphonia which negatively impacts daily life of patients. Current therapeutic options are limited due in large part to the high complexity of the micro-structure of the VF. Innovative therapies derived from adipose tissue such as stromal vascular fraction (SVF) or adipose derived stromal/ stem cells (ASC) are currently being evaluated in this indication and paracrine anti-fibrotic effects are considered as predominant mechanisms. Methods: The paracrine anti-fibrotic effects of SVF and ASC from healthy donors were tested in an innovative in vitro fibrogenesis model employing human VF fiboblasts (hVFF) and the principles of macromolecular crowding (MMC). Biosynthesis of collogen and alpha-smooth-muscle actin (αSMA) expression in hVFF were quantified after five days of indirect coculture with ASC or SVF using silver stain, western blot and RT-qPCR analysis. Results: Fibrogenesis was promoted by addition of transforming growth factor beta 1 (TGFβ1) combined with MMC characterized by an enhanced deposition of fibrillar collagens and the acquisition of a myofibroblast phenotype (overexpression of αSMA). Adipose-derived therapies led to a reduction in the αSMA expression and the collagen content was lower in hVFF co-cultivated with SVF. Discussion: ASC and SVF promoted significant prevention of fibrosis in an in vitro fibrogenesis model through paracrine mechanisms, supporting further development of adipose-derived cellular therapies in VF scarring.

show abstract

Establishing principles of macromolecular crowding for in vitro fibrosis research of the vocal fold lamina propria

Abstract: NA.

Cited by 17 publications

References 20 publications

Fibrillar fibronectin plays a key role as nucleator of collagen I polymerization during macromolecular crowding-enhanced matrix assembly

Fibrillar fibronectin plays a key role as nucleator of collagen I polymerization during macromolecular crowding-enhanced matrix assembly

Towards an in vitro fibrogenesis model of human vocal fold scarring

Paracrine Effects Of Adipose-Derived Stromal/Stem Cells And Stromal Vascular Fraction In An In Vitro Fibrogenesis Model Of Human Vocal Fold Scarring

Contact Info

Product

Resources

About