Cell Patterning with Mucin Biopolymers

Crouzier, Thomas; Jang, Hongchul; Ahn, Ji‐Young; Stocker, Roman; Ribbeck, Katharina

doi:10.1021/bm400447z

Cited by 36 publications

(41 citation statements)

References 49 publications

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“…This did not come as a surprise, as the materials carry similarities with other poorly cell-adherent materials such as alginate 42 and hyaluronic acid 43 that are also hydrated, negatively charged and do not carry any known binding ligands to integrins. This is also in agreement with previous reports of mucin coatings preventing cell adhesion 44,45 .…”

Section: Discussionsupporting

confidence: 94%

Mucin hydrogel glyco-modulation to investigate immune activity of mucin glycans

Yan

Hjorth

Winkeljann

et al. 2019

Preprint

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Mucins are multifunctional glycosylated proteins that are increasingly investigated as building blocks of novel biomaterials. Once assembled into hydrogels (Muc gels), mucins were shown to modulate the recruitment and activation of immune cells and avoid fibrous encapsulation in vivo.However, nothing is known about the early immune response to Muc gels. This study characterizes the response of macrophages, important orchestrators of the material-mediated immune response, over the first 7 days in contact with Muc gels. The role of mucin-bound sialic acid sugar residues was investigated by first enzymatically cleaving the sugar, then assembling the mucin variants into covalently crosslinked hydrogels with rheological and surface nanomechanical properties similar to non-modified Muc gels. Results with THP1 and human primary peripheral blood monocytesderived macrophages were strikingly consistent and showed that Muc gels transiently activate the expression of both pro-inflammatory and anti-inflammatory cytokines and cell surface markers, with a maximum on the first day and loss of the effect after 7 days. The activation was sialic aciddependent for a majority of the markers followed. The pattern of gene expression, protein expression, and functional measurements did not strictly correspond to M1 or M2 macrophage phenotypes. This study highlights the complex early events in macrophage activation in contact with mucin materials and the importance of sialic acid residues in such a response. The enzymatic glyco-modulation of Muc gels appears as a useful tool to help understand the biological functions of specific glycans on mucins which can further inform on their use in various biomedical applications.

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Section: Discussionsupporting

confidence: 94%

Mucin hydrogel glyco-modulation to investigate immune activity of mucin glycans

Yan

Hjorth

Winkeljann

et al. 2019

Preprint

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“…Mucin‐bound glycans within mucin coatings can contribute to the repulsion of mammalian cells . To examine if mucin‐bound glycans also play a role in bacterial repulsion, bacterial attachment was tested on coatings made from deglycosylated mucins, which are henceforth referred to as apo‐mucins.…”

Section: Resultsmentioning

confidence: 99%

Probing the Role of Mucin‐Bound Glycans in Bacterial Repulsion by Mucin Coatings

Crouzier

Ribbeck

2015

Adv Materials Inter

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in a planktonic state and impair biofi lm development. A similar effect has been observed for human salivary mucin MUC5B toward the bacterium Streptococcus mutans . [ 9 ] Moreover, cell-surface mucins can reduce Staphylococcus aureus attachment to corneal epithelial cells [ 10 ] and can limit Helicobacter pylori attachment to gastric epithelial cells. [ 11 ] Given their ability to suppress microbial surface attachment in native conditions, mucins have also been studied in the context of microbe-repelling coatings on polyacrylic acid, poly(methyl methacrylate), silicone, polyurethane, and polystyrene surfaces. [ 12,13 ] These studies show that it is diffi cult to predict how mucin coatings perform because their function depends on the surfaces to which they are adsorbed and on deposition conditions. For example, bovine submaxillary mucin coatings reduce C. albicans attachment to silicon surfaces, but increase its attachment to poly(acrylic acidb -methyl methacrylate) (PAAb -PMMA) surfaces. [ 13 ] This variability refl ects a gap in our understanding of which functional domains and biochemical properties of mucin coatings are important in mucin-microbe interactions.This study investigates which molecular components of mucins contribute to bacterial repulsion, specifi cally when coating polystyrene surfaces. The respiratory pathogens Streptococcus pneumoniae , S. aureus , and P. aeruginosa often colonize medical devices to cause infection, [ 2 ] and thus were examined here. Our data show that coatings made of gastric or submaxillary mucins effi ciently prevented surface attachment of S. pneumoniae and S. aureus , but not of P. aeruginosa . To dissect the role of mucin-bound glycans in bacterial repulsion, properties of native mucin coatings were compared to those of deglycosylated mucin coatings. We found that upon deglycosylation, mucin coatings changed in structure and lost their ability to repel bacteria, indicating that mucin-bound glycans are structural components capable of regulating surface attachment by S. pneumoniae and S. aureus . Results and Discussion Mucin Coatings Reduce Surface Attachment of S. pneumoniae and S. aureus , but Not of P. aeruginosaSurface attachment of three common respiratory pathogens ( S. pneumoniae , S. aureus , and P. aeruginosa ) to mucin coatings was evaluated using in-house purifi ed porcine gastric mucins Microbial colonization of implanted medical devices in humans can lead to device failure and life-threatening infections. One strategy to prevent this unwanted colonization is to coat devices with polymers that reduce bacterial attachment. This study investigates how mucins, a class of biopolymers found in mucus, can be used as surface coatings to prevent attachment of selected respiratory pathogens to polystyrene surfaces. Our data show that coatings of porcine gastric mucins or bovine submaxillary mucins reduce surface attachment by Streptococcus pneumoniae and Staphylococcus aureus , but not Pseudomonas aeruginosa . To elucidate how mucin coatings repel S. pneumoniae and S....

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“…This discrepancy of our and previous data might be explained by the different surface characteristics of the plates used in the assays. In our study, mucin was used to foster the biofilm formation (Roy et al 2011, 2012), since apart from facilitating initial attachment and promoting growth, coating of the plates with this glycoprotein also confers highly hydrophilic properties to the inherent hydrophobic polystyrene surfaces (Crouzier et al 2013) and simultaneously mirrors the native habitat of the bacterium. Thus, it is conceivable that a Δ wecC mutant (lacking three charged sugar residues) exhibits reduced biofilm formation when grown on hydrophilic surface, while the comparably more hydrophobic, non-coated polystyrene plates might have favored the opposite.…”

Section: Discussionmentioning

confidence: 99%

Tannerella forsythiastrains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

et al. 2016

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Tannerella forsythia is an anaerobic, Gram-negative periodontal pathogen. A unique O-linked oligosaccharide decorates the bacterium’s cell surface proteins and was shown to modulate the host immune response. In our study, we investigated the biosynthesis of the nonulosonic acid (NulO) present at the terminal position of this glycan. A bioinformatic analysis of T. forsythia genomes revealed a gene locus for the synthesis of pseudaminic acid (Pse) in the type strain ATCC 43037 while strains FDC 92A2 and UB4 possess a locus for the synthesis of legionaminic acid (Leg) instead. In contrast to the NulO in ATCC 43037, which has been previously identified as a Pse derivative (5-N-acetimidoyl-7-N-glyceroyl-3,5,7,9-tetradeoxy-l-glycero-l-manno-NulO), glycan analysis of strain UB4 performed in this study indicated a 350-Da, possibly N-glycolyl Leg (3,5,7,9-tetradeoxy-d-glycero-d-galacto-NulO) derivative with unknown C5,7 N-acyl moieties. We have expressed, purified and characterized enzymes of both NulO pathways to confirm these genes’ functions. Using capillary electrophoresis (CE), CE–mass spectrometry and NMR spectroscopy, our studies revealed that Pse biosynthesis in ATCC 43037 essentially follows the UDP-sugar route described in Helicobacter pylori, while the pathway in strain FDC 92A2 corresponds to Leg biosynthesis in Campylobacter jejuni involving GDP-sugar intermediates. To demonstrate that the NulO biosynthesis enzymes are functional in vivo, we created knockout mutants resulting in glycans lacking the respective NulO. Compared to the wild-type strains, the mutants exhibited significantly reduced biofilm formation on mucin-coated surfaces, suggestive of their involvement in host-pathogen interactions or host survival. This study contributes to understanding possible biological roles of bacterial NulOs.

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Cell Patterning with Mucin Biopolymers

Cited by 36 publications

References 49 publications

Mucin hydrogel glyco-modulation to investigate immune activity of mucin glycans

Mucin hydrogel glyco-modulation to investigate immune activity of mucin glycans

Probing the Role of Mucin‐Bound Glycans in Bacterial Repulsion by Mucin Coatings

Tannerella forsythiastrains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

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