Autoinducer Sensing Microarrays by Reporter Bacteria Encapsulated in Hybrid Supramolecular-Polysaccharide Hydrogels.

A generally applicable strategy to obtain mechanically robust hydrogels for the incorporation and containment of functional reporter bacteria for the microarray and microparticle-based detection and signaling of N-acyl homoserine lactone autoinducers (3OC12 HSL) at relevant concentrations is reported. For reinforcing hydrogels of 1,4-bi(phenylalanine-diglycol)-benzene (PDB), a hybrid hydrogel is formed by the combination of PDB self-assembly with Ca2+ mediated alginate crosslinking. The different assembly mechanisms are shown not to interfere with each other and despite the more than four-fold increased moduli of the hydrogels, diffusion of autoinducers into the gels remains efficient and Escherichia coli pLuxR-green fluorescent protein (GFP) reporter bacteria are proliferating. Templating affords reporter bacteria-loaded hydrogels with controllable shape and size. Upon exposure to 3OC12 HSL, the embedded bacteria exhibit an up to 12 ± 3 times increase in fluorescence intensity due to autoinducer-triggered GFP expression. This approach can serve as a potentially generally applicable strategy to sensitively detect bacteria via their secreted autoinducers.

[1]  M. L. Simpson,et al.  Stochastic Assembly of Bacteria in Microwell Arrays Reveals the Importance of Confinement in Community Development , 2016, PloS one.

[2]  K. Magnusson,et al.  The Pseudomonas aeruginosa N-Acylhomoserine Lactone Quorum Sensing Molecules Target IQGAP1 and Modulate Epithelial Cell Migration , 2012, PLoS pathogens.

[3]  A. Nakao,et al.  Drastic difference in porous structure of calcium alginate microspheres prepared with fresh or hydrolyzed sodium alginate. , 2011, Journal of colloid and interface science.

[4]  Di Zhang,et al.  Mechanical reinforcement of C2-phenyl-derived hydrogels for controlled cell adhesion , 2013 .

[5]  N. Thet,et al.  Photopolymerization of polydiacetylene in hybrid liposomes: effect of polymerization on stability and response to pathogenic bacterial toxins. , 2014, The journal of physical chemistry. B.

[6]  Xiaocen Dou,et al.  Amino Acids and Peptide‐Based Supramolecular Hydrogels for Three‐Dimensional Cell Culture , 2017, Advanced materials.

[7]  Chueh Loo Poh,et al.  Engineering microbes to sense and eradicate Pseudomonas aeruginosa, a human pathogen , 2011, Molecular systems biology.

[8]  S. Rokka,et al.  Protecting probiotic bacteria by microencapsulation: challenges for industrial applications , 2010 .

[9]  David W Williams,et al.  A review of the scientific evidence for biofilms in wounds , 2012, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[10]  S. Booth,et al.  Prototype Development of the Intelligent Hydrogel Wound Dressing and Its Efficacy in the Detection of Model Pathogenic Wound Biofilms. , 2016, ACS applied materials & interfaces.

[11]  H. Schönherr,et al.  Rapid Detection of Escherichia coli via Enzymatically Triggered Reactions in Self-Reporting Chitosan Hydrogels. , 2015, ACS applied materials & interfaces.

[12]  D. Mooney,et al.  Alginate: properties and biomedical applications. , 2012, Progress in polymer science.

[13]  D. Weitz,et al.  Composite alginate gels for tunable cellular microenvironment mechanics , 2016, Scientific Reports.

[14]  David W Williams,et al.  Detection and identification of specific bacteria in wound biofilms using peptide nucleic acid fluorescent in situ hybridization (PNA FISH). , 2009, Microbiology.

[15]  Hisatoshi Kobayashi,et al.  Bone regeneration on a collagen sponge self-assembled peptide-amphiphile nanofiber hybrid scaffold. , 2007, Tissue engineering.

[16]  Brian Ralph,et al.  Determination of the cation content of alginate thin films by FTi.r. spectroscopy , 1997 .

[17]  S. Kjelleberg,et al.  A novel and sensitive method for the quantification of N-3-oxoacyl homoserine lactones using gas chromatography-mass spectrometry: application to a model bacterial biofilm. , 2000, Environmental microbiology.

[18]  E. Kumacheva,et al.  Microfluidic generation of composite biopolymer microgels with tunable compositions and mechanical properties. , 2014, Biomacromolecules.

[19]  H. Schönherr,et al.  Dual Enzyme-Responsive Capsules of Hyaluronic Acid-block-Poly(Lactic Acid) for Sensing Bacterial Enzymes. , 2015, Macromolecular rapid communications.

[20]  E. Greenberg,et al.  A second N-acylhomoserine lactone signal produced by Pseudomonas aeruginosa. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  C. Melander,et al.  Biologically inspired strategies for combating bacterial biofilms. , 2013, Current opinion in pharmacology.

[22]  Jessica L. Terrell,et al.  Encapsulated fusion protein confers “sense and respond” activity to chitosan–alginate capsules to manipulate bacterial quorum sensing , 2013, Biotechnology and bioengineering.

[23]  Luca Gasperini,et al.  Natural polymers for the microencapsulation of cells , 2014, Journal of The Royal Society Interface.

[24]  Ali Khademhosseini,et al.  Microscale Strategies for Generating Cell-Encapsulating Hydrogels. , 2012, Polymers.

[25]  M. C. Feiters,et al.  Two-component dendritic gel: effect of stereochemistry on the supramolecular chiral assembly. , 2004, Chemistry.

[26]  Jie Zhou,et al.  Supramolecular biofunctional materials. , 2017, Biomaterials.

[27]  Shuguang Zhang.  Hydrogels: Wet or let die. , 2004, Nature materials.

[28]  David K. Smith,et al.  Enantioselective component selection in multicomponent supramolecular gels. , 2014, Journal of the American Chemical Society.

[29]  B. Ahmer.  Cell‐to‐cell signalling in Escherichia coli and Salmonella enterica , 2004, Molecular microbiology.

[30]  H. Gadebusch,et al.  In vitro antimicrobial activity of dimethylsulfoxide. , 1968, Applied microbiology.

[31]  Di Zhang,et al.  Control of three-dimensional cell adhesion by the chirality of nanofibers in hydrogels. , 2014, Angewandte Chemie.

[32]  A. Heeres,et al.  Responsive cyclohexane-based low-molecular-weight hydrogelators with modular architecture. , 2004, Angewandte Chemie.

[33]  Hang Lu,et al.  Microfluidic‐Based Generation of Size‐Controlled, Biofunctionalized Synthetic Polymer Microgels for Cell Encapsulation , 2014, Advanced materials.

[34]  Fast-Track, One-Step E. coli Detection: A Miniaturized Hydrogel Array Permits Specific Direct PCR and DNA Hybridization while Amplification. , 2016, Macromolecular bioscience.

[35]  Di Zhang,et al.  Gelator-polysaccharide hybrid hydrogel for selective and controllable dye release. , 2012, Journal of colloid and interface science.

[36]  Di Zhang,et al.  C2-symmetric benzene-based hydrogels with unique layered structures for controllable organic dye adsorption , 2012 .

[37]  Lai Wah Chan,et al.  Microencapsulation of microbial cells , 2013 .

[38]  Wai-Leung Ng,et al.  Specificity and complexity in bacterial quorum-sensing systems , 2016, FEMS microbiology reviews.

[39]  A. Jenkins,et al.  An electrochemical sensor concept for the detection of bacterial virulence factors from Staphylococcus aureus and Pseudomonas aeruginosa , 2015 .

[40]  Jie Zhou,et al.  Supramolecular Hydrogelators and Hydrogels: From Soft Matter to Molecular Biomaterials , 2015, Chemical reviews.

[41]  D. Mooney,et al.  Hydrogels for tissue engineering: scaffold design variables and applications. , 2003, Biomaterials.

[42]  G. Gokel,et al.  The aqueous medium-dimethylsulfoxide conundrum in biological studies , 2015 .

[43]  Wilfred Chen,et al.  Microbial Biosensors: Engineered Microorganisms as the Sensing Machinery , 2013, Sensors.

[44]  Tao Wang,et al.  Enzyme degradable polymersomes from hyaluronic acid-block-poly(ε-caprolactone) copolymers for the detection of enzymes of pathogenic bacteria. , 2015, Biomacromolecules.

[45]  Bonnie L. Bassler,et al.  Quorum sensing signal–response systems in Gram-negative bacteria , 2016, Nature Reviews Microbiology.