Effect of in vitro collagen fibrillogenesis on Langmuir-Blodgett (LB) deposition for cellular behavior regulation

AbstractThe extracellular matrix (ECM) is a complex mixture composed of fibrillar collagens as well as additional protein and carbohydrate components. Proteoglycans (PGs) contribute to the heterogeneity of the ECM and play an important role in its structure and function. While the small leucine rich proteoglycans (SLRPs), including decorin and lumican, have been studied extensively as mediators of collagen fibrillogenesis and organization, the function of large matrix PGs in collagen matrices is less well known. In this study, we showed that different matrix PGs have distinct roles in regulating collagen behaviors. We found that versican, a large chondroitin sulfate PG, promotes collagen fibrillogenesis in a turbidity assay and upregulates cell-mediated collagen compaction and reorganization, whereas aggrecan, a structurally-similar large PG, has different and often opposing effects on collagen. Compared to versican, decorin and lumican also have distinct functions in regulating collagen behaviors. The different ways in which matrix PGs interact with collagen have important implications for understanding the role of the ECM in diseases such as fibrosis and cancer, and suggest that matrix PGs are potential therapeutic targets.HighlightsSmall leucine rich proteoglycans (SLRPs) and large chondroitin sulfate (CS) proteoglycans (PGs) have distinct effects on collagen fibrous network behavior.Unlike other matrix proteoglycans, versican promotes collagen fibrillogenesis in an in vitro spectrophotometric (turbidity) assay.The versican core protein has a larger impact on collagen behavior in a fibrillogenesis assay than its glycosaminoglycan chains do.Versican increases the diameter of collagen fibers and the porosity of collagen fibrous networks, unlike aggrecan and SLRPs.The addition of versican to collagen does not alter fibroblast contractility but leads to enhanced cell-mediated collagen reorganization and contraction.

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An in-vitro assay using human spermatozoa to detect toxicity of biologically active substances

Scientific Reports ◽

10.1038/s41598-019-50929-z ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Tino Vollmer ◽

Börje Ljungberg ◽

Vera Jankowski ◽

Joachim Jankowski ◽

Griet Glorieux ◽

...

Keyword(s):

Ex Vivo ◽

Toxicity Testing ◽

Biologically Active ◽

Human Spermatozoa ◽

Vitro Assay ◽

Cellular Behavior ◽

Novel Method ◽

Set Up

Abstract Identifying the key toxic players within an in-vivo toxic syndrome is crucial to develop targeted therapies. Here, we established a novel method that characterizes the effect of single substances by means of an ex-vivo incubation set-up. We found that primary human spermatozoa elicit a distinct motile response on a (uremic) toxic milieu. Specifically, this approach describes the influence of a bulk toxic environment (uremia) as well as single substances (uremic toxins) by real-time analyzing motile cellular behavior. We established the human spermatozoa-based toxicity testing (HSTT) for detecting single substance-induced toxicity to be used as a screening tool to identify in-vivo toxins. Further, we propose an application of the HSTT as a method of clinical use to evaluate toxin-removing interventions (hemodialysis).

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Enzymatically degradable alginate/gelatin bioink promotes cellular behavior and degradation in vitro and in vivo

Biofabrication ◽

10.1088/1758-5090/ab38ef ◽

2019 ◽

Vol 11 (4) ◽

pp. 045020 ◽

Cited By ~ 6

Author(s):

Bin Yao ◽

Tian Hu ◽

Xiaoliang Cui ◽

Wei Song ◽

Xiaobing Fu ◽

...

Keyword(s):

Cellular Behavior

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Integration of cell cycle signals by multi-PAS domain kinases

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1808543115 ◽

2018 ◽

Vol 115 (30) ◽

pp. E7166-E7173 ◽

Cited By ~ 8

Author(s):

Thomas H. Mann ◽

Lucy Shapiro

Keyword(s):

Intracellular Signaling ◽

Spatial Information ◽

Caulobacter Crescentus ◽

Master Regulator ◽

Cellular Behavior ◽

Cell Pole ◽

Spatial Niche ◽

Spatiotemporal Control ◽

Stimulation Of

Spatial control of intracellular signaling relies on signaling proteins sensing their subcellular environment. In many cases, a large number of upstream signals are funneled to a master regulator of cellular behavior, but it remains unclear how individual proteins can rapidly integrate a complex array of signals within the appropriate spatial niche within the cell. As a model for how subcellular spatial information can control signaling activity, we have reconstituted the cell pole-specific control of the master regulator kinase/phosphatase CckA from the asymmetrically dividing bacterium Caulobacter crescentus. CckA is active as a kinase only when it accumulates within a microdomain at the new cell pole, where it colocalizes with the pseudokinase DivL. Both proteins contain multiple PAS domains, a multifunctional class of sensory domains present across the kingdoms of life. Here, we show that CckA uses its PAS domains to integrate information from DivL and its own oligomerization state to control the balance of its kinase and phosphatase activities. We reconstituted the DivL–CckA complex on liposomes in vitro and found that DivL directly controls the CckA kinase/phosphatase switch, and that stimulation of either CckA catalytic activity depends on the second of its two PAS domains. We further show that CckA oligomerizes through a multidomain interaction that is critical for stimulation of kinase activity by DivL, while DivL stimulation of CckA phosphatase activity is independent of CckA homooligomerization. Our results broadly demonstrate how signaling factors can leverage information from their subcellular niche to drive spatiotemporal control of cell signaling.

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Potential modifier role of the R618Q variant of proα2(I)collagen in type I collagen fibrillogenesis: in vitro assembly analysis

Molecular Genetics and Metabolism ◽

10.1016/j.ymgme.2004.03.004 ◽

2004 ◽

Vol 82 (2) ◽

pp. 144-153 ◽

Cited By ~ 5

Author(s):

Anthony N Vomund ◽

Stephen R Braddock ◽

Gary F Krause ◽

Charlotte L Phillips

Keyword(s):

Type I Collagen ◽

Type I ◽

Collagen Fibrillogenesis ◽

In Vitro Assembly ◽

Assembly Analysis

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Influence of manganese ions on cellular behavior of human osteoblasts in vitro

Biomolecular Engineering ◽

10.1016/j.bioeng.2007.08.003 ◽

2007 ◽

Vol 24 (5) ◽

pp. 531-536 ◽

Cited By ~ 35

Author(s):

Frank Lüthen ◽

Ulrike Bulnheim ◽

Petra D. Müller ◽

Joachim Rychly ◽

Henrike Jesswein ◽

...

Keyword(s):

Manganese Ions ◽

Human Osteoblasts ◽

Cellular Behavior

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Morphometry of cellular behavior of coelomocytes from starfish Asterias amurensis

PeerJ ◽

10.7717/peerj.12514 ◽

2021 ◽

Vol 9 ◽

pp. e12514

Author(s):

Yuriy A. Karetin

Keyword(s):

Cell Morphology ◽

Marine Invertebrate ◽

Cell Types ◽

Cell Behavior ◽

Asterias Amurensis ◽

Nonlinear Parameters ◽

Cellular Behavior ◽

Wide Range ◽

Third Stage

A comprehensive statistical analysis using a wide range of linear and non-linear morphological parameters enabled identification of the main stages in the in vitro dynamics of cell behavior of immune cells of the marine invertebrate Asterias amurensis (Echinodermata, Asteroidea). Three stages may be distinguished in the cell behavior, which are characterized by the differences in complexity of the cell boundary microsculpture as well as by the size and asymmetry of the cell and convex hull of the cell. The first stage (5 min after placing cells onto a substrate) is characterized by more complex cell morphology and an increase in the process number and spreading area. The second stage (15 min) is characterized by simplification of cell morphology, retraction of some processes, and rounding of cells upon continued cell spreading. At the third stage (60 min), new large processes with rounded contours emerge due to partial retraction of the flattened cell surface. Each stage is characterized by statistically significant differences in several linear and nonlinear parameters of the external morphology for all cell types.

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