Bacteria-surface interaction in the presence of proteins and surface attached poly(ethylene glycol) methacrylate chains

This study analyzes the adhesion behavior of the gram positive bacteria, Staphylococcus aureus (S. aureus), and the gram negative bacteria, Escherichia coli (E. coli), on polypyrrole (PPY) surfaces in the presence of poly(ethylene glycol) methacrylate (PEGMA) chains and plasma proteins (bovine serum albumin and bovine plasma fibrinogen) either preadsorbed on the film surface or in the bacterial suspension. Bacterial adhesion experiments performed in a suspension of bacterial cells and protein may give important insights on the behavior of bacterial adhesion in an in vivo environment. Protein adsorption and bacterial adhesion on PEGMA-grafted PPY films were reduced by about a factor of 2-4 compared with those on the pristine PPY films. In addition, the number of bacterial cells adhering on the substrate is dependent not only on the type of protein present, but also the sequence of exposure to the protein relative to the bacteria. Furthermore, bacteria-surface adhesion force was measured using the atomic force microscopy with increasing lateral force to detach the individual cell. The adhesion force of S. aureus is influenced by PEGMA and plasma protein modification and is significantly higher than that of E. coli for all substrates tested. The number of adherent cells on the substrate is shown to be directly correlated to the bacterial adhesion force.     

Reduced medical infection related bacterial strains adhesion on bioactive RGD modified titanium surfaces: a first step toward cell selective surfaces

Ideally, implants should inhibit nonspecific protein adsorption, bacterial adhesion, and at the same time, depending on the final application be selective toward cellular adhesion and spreading for all or only selected cell types. Poly(L-lysine)-grafted-poly(ethylene glycol) (PLL-g-PEG) polymers have been shown to adsorb from aqueous solution onto negatively charged metal oxide surfaces, reducing protein adsorption as well as fibroblast, osteoblast and epithelial cell adhesion significantly. PLL-g-PEG can be functionalized with bioligands such as RGD (Arg-Gly-Asp), which then restores host cell adhesion, but the surface remains resistant to nonspecific protein adsorption. Previously, it was also shown that both nonfunctionalized PLL-g-PEG and RGD-peptide functionalized PLL-g-PEG reduced the adhesion of Staphylococcus aureus to titanium (Ti) surfaces. The present study looked at the effect of other implant associated infection relevant bacteria, Staphylococcus epidermidis, Streptococcus mutans and Pseudomonas aeruginosa towards the same surface chemistries. The different surfaces were exposed to the bacteria for 1-24 h, and bacteria surface density was evaluated using scanning electron microscopy (SEM) and fluorescence light microscopy (FM). The adhesion of all bacteria strains tested was reduced on Ti surfaces coated with PLL-g-PEG compared to uncoated Ti surfaces even in the presence of RGD. The percentage reduction in bacterial adhesion over the 24-h culture time investigated was 88%-98%, depending on the bacteria type. Therefore, coating surfaces with PLL-g-PEG/PEG-RGD allows cells such as fibroblasts and osteoblasts to attach but not bacteria, resulting in a selective biointeractive pattern that may be useful on medical implants.     

Manipulation of bacterial adhesion and proliferation by surface charges of electrically polarized hydroxyapatite

The manipulation of bacterial adhesion and proliferation by surface charges built onto the surfaces of electrically polarized bioceramic hydroxyapatite (HAp) was investigated. The gram-positive bacteria Staphylococcus aureus (S. aureus) and the gram-negative bacteria Escherichia coli (E. coli) were cultivated on negatively charged, positively charged, and non-charged HAp surfaces (denoted as N-, P-, and 0-surface, respectively). The electrostatic force caused by the surface charges experimentally was proven to affect both adhesion and proliferation. Compared with the 0-surface of HAp ceramics over a 3-h cultivation, the population of adhered bacteria rapidly multiplied on the N-surface whereas it multiplied quite slowly on the P-surface. Compared with the 0-surface over a cultivation period of 12 to 72 h, the proliferation rate of the bacterial cell density per colony was accelerated on the N-surface and decelerated on the P-surface. The above results are attributed (1) to the electrostatic interaction between the cell surfaces and the charged surfaces of the polarized HAp, (2) to the stimulus of the electrostatic force for bacterial cells, and (3) to the concentration of the nutrient for the bacteria.     

Staphylococcus aureus adhesion to titanium oxide surfaces coated with non-functionalized and peptide-functionalized poly(L-lysine)-grafted-poly(ethylene glycol) copolymers

Implanted biomaterials are coated immediately with host plasma constituents, including extracellular matrix (ECM); this reaction may be undesirable in some cases. Poly(L-lysine)-grafted-poly(ethylene glycol) (PLL-g-PEG) has been shown to spontaneously adsorb from aqueous solution onto metal oxide surfaces, effectively reducing the degree of non-specific adsorption of blood and ECM proteins, and decreasing the adhesion of fibroblastic and osteoblastic cells to the coated surfaces. Cell adhesion through specific peptide-integrin receptors could be restored on surfaces coated with PLL-g-PEG functionalized with peptides of the RGD (Arg-Asp-Gly) type. To date, no study has examined the effect of surface modifications by PLL-g-PEG-based polymers on bacterial adhesion. The ability of Staphylococcus aureus to adhere to the ECM and plasma proteins deposited on biomaterials is a significant factor in the pathogenesis of medical-device-related infections. This study describes methods for visualizing and quantifying the adhesion of S. aureus to smooth and rough (chemically etched) titanium surfaces without and with monomolecular coatings of PLL-g-PEG, PLL-g-PEG/PEG-RGD and PLL-g-PEG/PEG-RDG. The different surfaces were exposed to S. aureus cultures for 1-24h and bacteria surface density was evaluated using scanning electron microscopy and fluorescence microscopy. Coating titanium surfaces with any of the three types of copolymers significantly decreased the adhesion of S. aureus to the surfaces by 89-93% for PLL-g-PEG, and 69% for PLL-g-PEG/PEG-RGD. Therefore, surfaces coated with PLL-g-PEG/PEG-RGD have the ability to attach cells such as fibroblasts and osteoblasts while showing reduced S. aureus adhesion, resulting in a selective biointeraction pattern that may be useful for applications in the area of osteosynthesis, orthopaedic and dental implantology.     

Reduced bacterial adhesion to fibrinogen-coated substrates via nitric oxide release

The ability of nitric oxide (NO)-releasing xerogels to reduce fibrinogen-mediated adhesion of Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli is described. A negative correlation was observed between NO surface flux and bacterial adhesion for each species tested. For S. aureus and E. coli, reduced adhesion correlated directly with NO flux from 0 to 30pmolcm(-2)s(-1). A similar dependence for S. epidermidis was evident from 18 to 30pmolcm(-2)s(-1). At a NO flux of 30pmolcm(-2)s(-1), surface coverage of S. aureus, S. epidermidis, and E. coli was reduced by 96, 48, and 88%, respectively, compared to non-NO-releasing controls. Polymeric NO release was thus demonstrated to be an effective approach for significantly reducing fibrinogen-mediated adhesion of both gram-positive and gram-negative bacteria in vitro, thereby illustrating the advantage of active NO release as a strategy for inhibiting bacterial adhesion in the presence of pre-adsorbed protein.     

Comparison of bacterial and tissue cell initial adhesion on hydrophilic/hydrophobic biomaterials

In this study, interactions of widely-used polymeric biomaterials, i.e. poly(hydroxyethyl methacrylate) (PHEMA) and its copolymer with dimethylaminoethyl methacrylate (PHEMA-20% DMAEMA), polyurethane (PU), polypropylene (PP), poly(vinyl chloride) (PVC), and poly(lactide-glycolide) (PLGA), with three pathogenic bacteria and one nonpathogen were investigated comparatively with the adhesion of two tissue cells in different morphologies, i.e. fibroblast-like baby hamster kidney (BHK 21) cells and epithelial Madine Darby kidney (MDBK) cells. Biomaterials were prepared in the membrane form by bulk polymerization or solvent casting. Surface characterization studies showed that these polymers have different surface free energies in the range of 26.9-63.1 erg cm(-2) and they have smooth surfaces. The bacteria used were; Escherichia coli ATCC 25922, Staphylococcus epidermidis ATCC 12228, Staphylococcus aureus, and Lactobacillus acidophilus B-13. Initial adhesion of bacteria to the polymeric surfaces was examined under static conditions and in a laminar flow cell. The adhesion behaviour of S. aureus and S. epidermidis was found independent of the polymeric surface hydrophobicity. However, the percentage of attached E. coli decreased when increasing the surface free energy of the polymer, while L. acidophilus showed just the opposite behaviour. The comparative results indicated that the adhesion of BHK and MDBK cell was lowest on the most hydrophilic PHEMA surface and highest on the most hydrophobic PP surface. In contrast to the case of bacterial adhesion, no relationship was found between polymer hydrophobicity and mammalian cell adherence.     

Bacterial adhesion and osteoblast function on titanium with surface-grafted chitosan and immobilized RGD peptide

Biomaterials-associated infections remain a source of serious complications in modern medicine. When a biomaterial is implanted in the body, the result of successful tissue integration or implant infection depends on the race for the surface between bacteria and tissue cells. One promising strategy to reduce the incidence of infection is the functionalization of the biomaterial surface to inhibit bacterial adhesion and encourage the growth of cells. In this in vitro study, the surface of titanium alloy substrates was first functionalized by covalently grafted chitosan (CS). The cell-adhesive Arg-Gly-Asp (RGD) peptide was then immobilized on the CS-grafted surface through covalent binding of peptide to the free NH(2) groups of CS. Both these functionalized surfaces showed a decrease in adhesion of Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis) compared with the pristine substrate. A significant increase in osteoblast cell attachment, proliferation, and alkaline phosphatase activity was observed on the surface with the immobilized Arg-Gly-Asp peptide. Thus, utilizing surface-grafted chitosan in conjunction with the cell-adhesive peptide to modify the metal surface provides a promising means for enhancing tissue integration of implants by reducing bacterial adhesion and promoting osteoblast functions.     

Identification of a nonfimbrial adhesive factor of an enterotoxigenic Escherichia coli strain.

An enterotoxigenic Escherichia coli strain (strain 2230), isolated from a patient with acute infantile diarrhea, was found to adhere only to the brush border of human intestinal epithelial cells. This strain does not hemagglutinate human, bovine, chicken, or guinea pig erythrocytes. The adhesion of E. coli 2230 appears to be mediated by a nonfimbrial bacterial surface protein of 16,000 daltons which can be extracted by heating the bacteria at 60 degrees C for 20 min. This surface protein is implicated as an adhesive factor because pretreatment of enterocytes with this protein extract completely inhibits the adhesion of E. coli 2230. This adhesive factor is serologically distinct from other adhesive factors found in enterotoxigenic E. coli strains. A plasmid DNA of 66 megadaltons is involved in the synthesis of this nonfimbrial adhesive factor.     

Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection

Staphylococcus aureus and Escherichia coli are among the most prevalent species of gram-positive and gram-negative bacteria, respectively, that induce clinical mastitis. The innate immune system comprises the immediate host defense mechanisms to protect against infection and contributes to the initial detection of and proinflammatory response to infectious pathogens. The objective of the present study was to characterize the different innate immune responses to experimental intramammary infection with E. coli and S. aureus during clinical mastitis. The cytokine response and changes in the levels of soluble CD14 (sCD14) and lipopolysaccharide-binding protein (LBP), two proteins that contribute to host recognition of bacterial cell wall products, were studied. Intramammary infection with either E. coli or S. aureus elicited systemic changes, including decreased milk output, a febrile response, and induction of the acute-phase synthesis of LBP. Infection with either bacterium resulted in increased levels of interleukin 1beta (IL-1beta), gamma interferon, IL-12, sCD14, and LBP in milk. High levels of the complement cleavage product C5a and the anti-inflammatory cytokine IL-10 were detected at several time points following E. coli infection, whereas S. aureus infection elicited a slight but detectable increase in these mediators at a single time point. Increases in IL-8 and tumor necrosis factor alpha were observed only in quarters infected with E. coli. Together, these data demonstrate the variability of the host innate immune response to E. coli and S. aureus and suggest that the limited cytokine response to S. aureus may contribute to the well-known ability of the bacterium to establish chronic intramammary infection.     

Effect of bacterial competition on the opsonization, phagocytosis, and intracellular killing of microorganisms by granulocytes.

The ingestion of Escherichia coli by human granulocytes in vitro was reduced in the presence of Bacteroides fragilis or Staphylococcus aureus. This reduction of ingestion proved to be mainly attributable to the absence of opsonization of E. coli, which was due to complement consumption by B. fragilis and S. aureus. The intracellular killing of E. coli was decreased in the presence of B. fragilis and S. aureus because of consumption of complement components required for extracellular stimulation of granulocytes to kill intracellular bacteria. Decreased intracellular killing of E. coli by granulocytes containing either B. fragilis or S. aureus is due to the limited killing capacity of granulocytes. These interactions between E. coli and B. fragilis or S. aureus found for phagocytosis and intracellular killing were also observed in in vivo studies: in an experimental thigh lesion infection in mice, E. coli showed stronger proliferation after coinoculation with B. fragilis or with S. aureus than after injection of E. coli alone. These in vitro and in vivo findings indicate that bacterial interactions, not only between aerobic and anaerobic bacteria but also between two species of aerobic microorganisms, compete for host defense mechanisms (i.e., opsonization, phagocytosis, and intracellular killing).     

Effects of multiplicity of infection, bacterial protein synthesis, and growth phase on adhesion to and invasion of human cell lines by Salmonella typhimurium.

Monolayers of intestine 407 (Int-407) cells were infected with the virulent Salmonella typhimurium strain C52, and the adhesion to and invasion of these cells were studied. The effects of the multiplicity of infection and growth phase of the bacteria (logarithmic versus stationary) on the interaction with eukaryotic cells were investigated. In contrast to other reports, we found no differences in the adhesive and invasive capacities of bacteria derived from logarithmic- or stationary-phase cultures. Invasion by S. typhimurium required bacterial protein synthesis and live Int-407 cells. Bacteria adhered equally well to dead or live Int-407 cells, which indicates that adhesion does not require metabolically active cells. Adhesion of S. typhimurium followed saturation kinetics, with a maximum of 10 adhesive bacteria per cell. This indicates that there is a limited number of bacterial adhesion sites (receptors) available on the surface of the host cell. Killed and live bacteria adhered equally well and competed with each other for cellular adhesion sites. This and adhesion experiments performed in the presence of chloramphenicol showed that bacterial protein synthesis is not required for adhesion. The general validity of the results obtained with S. typhimurium C52 was confirmed by comparing the invasion and adhesion data with those of the frequently used SL1344 and SR11 strains. In addition, we assayed the adhesion and invasion of S. typhimurium C52, SL1344, and SR11 and 27 S. typhimurium field isolates with Int-407, HeLa, and HEp-2 cells.     

Curcumin suppresses Streptococcus mutans adherence to human tooth surfaces and extracellular matrix proteins

Streptococcus mutans is the key causative agent of caries and infective endocarditis. The first step in biofilm development and the consequent initiation of further disease is bacterial adherence to host cell surfaces. Therefore, the aim of this study was to evaluate the inhibitory effect of curcumin on S. mutans adherence to extracellular matrices and tooth surfaces. The effect of curcumin on the ability of S. mutans to adhere to glass surfaces coated with collagen and fibronectin was tested in order to determine whether the decrease of the bacterial adhesion by curcumin is achieved by hindering the bacteria in adhering to collagen and/or fibronectin. Also, human teeth inoculated with S. mutans were treated with curcumin in vitro in order to assess the relevance of the anti-adhesive effect to oral conditions in vivo. The minimum inhibitory concentration (MIC) at which curcumin completely inhibited bacterial growth was 128?μg/mL. The addition of curcumin below the MIC diminished bacterial adherence onto both collagen- and fibronectin-coated glass surfaces and human tooth surfaces. It appears that the anti-adhesive effect of curcumin against S. mutans is mediated through collagen and fibronectin. These results support the widespread use of curcumin as a food-based antimicrobial agent.     

Bacterial adhesion onto azidated poly(vinyl chloride) surfaces

A plasticized poly(vinyl chloride) surface was modified by azidation using sodium azide in the presence of a phase transfer catalyst in aqueous media. Subsequent to azidation, the surface was crosslinked using ultraviolet radiation. Contact angle measurements showed that the surface became hydrophilic on azidation whereas photoirradiation did not have any further effect on the hydrophilicity of the azidated surface. Control, azidated, and photocrosslinked surfaces were exposed to two strains of bacteria commonly implicated in device infection such as Staphylococcus aureus and Escherichia coli. Whereas the control and photocrosslinked surfaces showed no significant difference in bacterial adhesion, the azidated surface showed significantly reduced adhesion to both strains. Data obtained indicate that the presence of an intact azide function on the polymer surface is responsible for the reduced bacterial adherence and the surface hydrophobicity/hydrophilicity did not exert any effect in the present case. Although azides are known to be effective only against Gram-negative species, surprising was the observation that the azidated polymer surface was equally effective against a Gram-positive species such as S. aureus. Because sodium azide is routinely used as a preservative to prevent bacterial and fungal growth in many microbiology reagents and diagnostic kits, covalent binding of the azide onto a polymer surface or synthesizing azide containing polymers may be an interesting method to investigate in tackling the problem of bacterial adhesion and colonization of medical devices.     

Possible involvement of altered RGD sequence in reduced adhesive and spreading activities of advanced glycation end product-modified fibronectin to vascular smooth muscle cells

Although fibronectin (FN) modified by advanced glycation end products (AGEs) has been shown to contribute to the development of diabetic vascular complications through its reduced adhesive activity to vascular cells, little is known about changes in the cell binding domain of AGE-modified FN. Here we examined the mechanism of reduced adhesive and spreading activities of AGE-modified FN to vascular smooth muscle cells (SMCs), particularly the contribution of modification of Arg-Gly-Asp (RGD) sequence. Incubation with glucose caused not only the formation of N(epsilon) -carboxymethyllysine and pentosidine, but also polymerization of FN in a dose- and time-dependent manner. AGE-modified FN had significantly low adhesive and spreading activities to cultured SMCs. On the other hand, multimeric FN formed by disulfide bonds did not show any effect on either cell adhesion or spreading. The adhesive activity of type I collagen, one of the RGD sequence-containing proteins, to SMCs also decreased by AGE-modification. The inhibitory effect of AGE-modification on cell adhesion was significantly greater in type I collagen than in FN. Although the extent of AGE-modification of type I collagen was indistinguishable from that of FN, AGE-modification decreased the arginine content of type I collagen by 69.5% and of FN by 30.6%, compared with their non-glycated forms. The addition of RGD peptides caused a decrease in adhesion of SMCs to non-glycated FN, but not to AGE-modified FN. Modification of RGD sequence with glyoxal eliminated its inhibitory effect on cell adhesion. Our results suggest that a marked decrease in adhesive and spreading activities of AGE-modified FN to SMCs might largely be due to a modification of its RGD sequence by AGE, thus suggesting a potential link between AGE modification of FN and the pathogenesis of diabetic angiopathy.     

Possible Involvement of Altered RGD Sequence in Reduced Adhesive and Spreading Activities of Advanced Glycation End Product-Modified Fibronectin to Vascular Smooth Muscle Cells

Although fibronectin (FN) modified by advanced glycation end products (AGEs) has been shown to contribute to the development of diabetic vascular complications through its reduced adhesive activity to vascular cells, little is known about changes in the cell binding domain of AGE-modified FN. Here we examined the mechanism of reduced adhesive and spreading activities of AGE-modified FN to vascular smooth muscle cells (SMCs), particularly the contribution of modification of Arg-Gly-Asp (RGD) sequence. Incubation with glucose caused not only the formation of N-carboxymethyllysine and pentosidine, but also polymerization of FN in a dose- and time-dependent manner. AGE-modified FN had significantly low adhesive and spreading activities to cultured SMCs. On the other hand, multimeric FN formed by disulfide bonds did not show any effect on either cell adhesion or spreading. The adhesive activity of type I collagen, one of the RGD sequence-containing proteins, to SMCs also decreased by AGE-modification. The inhibitory effect of AGE-modification on cell adhesion was significantly greater in type I collagen than in FN. Although the extent of AGE-modification of type I collagen was indistinguishable from that of FN, AGE-modification decreased the arginine content of type I collagen by 69.5% and of FN by 30.6%, compared with their non-glycated forms. The addition of RGD peptides caused a decrease in adhesion of SMCs to non-glycated FN, but not to AGE-modified FN. Modification of RGD sequence with glyoxal eliminated its inhibitory effect on cell adhesion. Our results suggest that a marked decrease in adhesive and spreading activities of AGE-modified FN to SMCs might largely be due to a modification of its RGD sequence by AGE, thus suggesting a potential link between AGE modification of FN and the pathogenesis of diabetic angiopathy.     

Chitinase 3-like-1 enhances bacterial adhesion to colonic epithelial cells through the interaction with bacterial chitin-binding protein

Dysregulated host/microbial interactions play a pivotal role in the pathogenesis of inflammatory bowel disease. We previously reported that chitinase 3-like-1 (CHI3L1) enhances bacterial adhesion and invasion on/into colonic epithelial cells (CECs). In this study, we designed to identify the exact mechanism of how CHI3L1 enhances the bacterial adhesion on CECs in vitro. As compared with wild type (WT) of Serratia marcescens, chitin binding protein (CBP) 21 knockout strain of S. marcescens significantly decreased the adhesion to SW480 cells that express CHI3L1 endogenously. A CBP21 fusion protein was produced with CBP21-expressing vector, which was transformed into BL21 strain of Escherichia coli. CBP21 overexpression significantly increased the adhesion, but not invasion, of nonpathogenic E. coli. The adhesion of S. marcescens and CBP21-overexpressing E. coli was inhibited by coculture with chitin, but not with other carbohydrates. After overexpressing CHI3L1 on SW480 cells, the adhesion rate of CBP21-overexpressing E. coli was further increased by approximately twofold. Genetically engineered E. coli with a single mutation of either Thy-54 or Glu-55 position of CBP21 exhibited a decreased binding ability, and the binding was 74% diminished by the combined mutations of three amino acids (Thy-54, Glu-55 and Glu-60) as compared with WT. Inhibition of CHI3L1 by anti-CHI3L1 antibody or CHI3L1-specific short interfering RNA reduced the adhesion of CBP21-overexpressing E. coli to CECs. In conclusion, CHI3L1 is involved in the enhancement of CBP-expressing bacterial adhesion to CECs. CBP21 and its homologs may be required for the CHI3L1-mediated enhancement of bacterial adhesion to CECs through the conserved amino-acid residues.     

Rat polyvinyl sponge model for the study of infections: host factors and microbial proliferation.

Female rats were treated with several administration regimens of methylprednisolone, cobra venom anti-complementary factor, and cyclophosphamide in conjunction with polyvinyl sponge implantations. The effect of these drugs on host factors active against bacteria was evaluated with Staphylococcus aureus ATCC 25933, Escherichia coli K-12, and Pseudomonas aeruginosa CDC 7725. One of two implants in each animal was infected with 10(8) of one of the three bacteria, and bacterial and granulocyte content was determined in the infected and control sponges after 48 h. The single large dose of methylprednisolone decreased staphylococcal and E. coli clearance while promoting dissemination of P. aeruginosa. A low chronic dose of the steroid inhibited E. coli chemotaxis only. A higher dose of the steroid administered chronically interfered markedly with S. aureus and E. coli curtailment by the host while leading to enhanced dissemination of P. aeruginosa, accompanied by a precipitous decline in granulocytes. Results with cobra factor resembled the higher chronic dose of steroid enhancing, especially the dissemination of the pseudomonad and its anti-granulocytic propensity. Cyclophosphamide depression of granulocytes revealed the rat's ability to curtail the proliferation of particular S. aureus and E.coli strains even in the absence of leukocytes. This treatment resulted in the rapid spread of P. aeruginosa, leading to the death of some experimental animals. These experiments underline the versatility of this animal model in the study of host and microbial factors influential in infectious disease.     

Engineering hydrophobin DewA to generate surfaces that enhance adhesion of human but not bacterial cells

Hydrophobins are fungal proteins with the ability to form immunologically inert membranes of high stability, properties that makes them attractive candidates for orthopaedic implant coatings. Cell adhesion on the surface of such implants is necessary for better integration with the neighbouring tissue; however, hydrophobin surfaces do not mediate cell adhesion. The aim of this project was therefore to investigate whether the class I hydrophobin DewA from Aspergillus nidulans can be functionalized for use on orthopaedic implant surfaces. DewA variants bearing either one RGD sequence or the laminin globular domain LG3 binding motif were engineered. The surfaces of both variants showed significantly increased adhesion of mesenchymal stem cells (MSCs), osteoblasts, fibroblasts and chondrocytes; in contrast, the insertion of binding motifs RGD and LG3 in DewA did not increase Staphylococcus aureus adhesion to the hydrophobin surfaces. Proliferation of MSCs and their osteogenic, chondrogenic and adipogenic differentiation potential were not affected on these surfaces. The engineered surfaces therefore enhanced MSC adhesion without interfering with their functionality or leading to increased risk of bacterial infection.     

Targeted cell adhesion on selectively micropatterned polymer arrays on a poly(dimethylsiloxane) surface

Herein, we introduce the fabrication of polymer micropattern arrays on a chemically inert poly(dimethylsiloxane) (PDMS) surface and employ them for the selective adhesion of cells. To fabricate the micropattern arrays, a mercapto-ester—based photocurable adhesive was coated onto a mercaptosilane—coated PDMS surface and photopolymerized using a photomask to obtain patterned arrays at the microscale level. Robust polymer patterns, 380 μm in diameter, were successfully fabricated onto a PDMS surface, and cells were selectively targeted toward the patterned regions. Next, the performance of the cell adhesion was observed by anchoring cell adhesive linker, an RGD oligopeptide, on the surface of the mercapto-ester—based adhesive-cured layer. The successful anchoring of the RGD linker was confirmed through various surface characterizations such as water contact angle measurement, XPS analysis, FT-IR analysis, and AFM measurement. The micropatterning of a photocurable adhesive onto a PDMS surface can provide high structural rigidity, a highly–adhesive surface, and a physical pathway for selective cell adhesion, while the incorporated polymer micropattern arrays inside a PDMS microfluidic device can serve as a microfluidic platform for disease diagnoses and high-throughput drug screening.