Higher Cell Adhesion Research Articles

Controlled Shear Flow Directs Osteogenesis on UHMWPE-Based Hybrid Nanobiocomposites in a Custom-Designed PMMA Microfluidic Device.

The combinatorial influence of a biophysical cue (substrate stiffness) and biomechanical cue (shear flow) on the osteogenesis modulation of human mesenchymal stem cells (hMSCs) is studied for bone regenerative applications. In this work, we report stem cell differentiation on an ultra high molecular weight polyethylene (UHMWPE)-based hybrid nanobiocomposite [reinforced with a multiwalled carbon nanotube (MWCNT) and/or nanohydroxyapatite (nHA)] under a physiologically relevant shear flow (1 Pa) in a custom-built microfluidic device. Using a genotypic assessment with qRT-PCR and phenotypic assessment through analysis of cytoskeletal remodelling and marker proteins, the role of shear on the progression of osteogenesis modulation has been quantitatively established with statistically significant differences between nHA-reinforced and MWCNT-reinforced UHMWPE. Early-stage (alkaline phosphatase activity at day 8), middle-stage (matrix collagenation at day 14), and late-stage (matrix calcification at day 20) events were analyzed using mRNA expression changes of a limited cell volume after microfluidic culture experiments. The conventional Petri dish culture (static) exhibited an increased osteogenesis for nanoparticle-reinforced UHMWPE, irrespective of the type of nanoparticle. The shear-mediated culture experiments resulted in noticeable differences in the degree of osteogenesis with MWCNT being more effective than nHA reinforcement. The shear-mediated osteogenesis has been attributed to the skewed cellular morphology with a higher cell adhesion (vinculin expression) on UHMWPE and nHA than that of UHMWPE and MWCNT. The signatures of the cytoskeletal changes are reflected in terms of left-to-right (L-R) chirality as well as alignment and pattern of actin fibers. Moreover, stemness (vimentin expression) was found to be decreased because of differentiation. The electrophysiological analysis using patch clamp experiments also revealed a higher inward calcium current and intracellular calcium activity for the cells grown on the UHMWPE and nHA nanobiocomposite under shear. Overall, the present study conclusively establishes the synergistic role of substrate stiffness and shear on osteogenesis of hMSCs, in vitro.

Comparison of Six Different Silicones In Vitro for Application as Glaucoma Drainage Device.

Silicones are widely used in medical applications. In ophthalmology, glaucoma drainage devices are utilized if conservative therapies are not applicable or have failed. Long-term success of these devices is limited by failure to control intraocular pressure due to fibrous encapsulation. Therefore, different medical approved silicones were tested in vitro for cell adhesion, cell proliferation and viability of human Sclera (hSF) and human Tenon fibroblasts (hTF). The silicones were analysed also depending on the sample preparation according to the manufacturer’s instructions. The surface quality was characterized with environmental scanning electron microscope (ESEM) and water contact angle measurements. All silicones showed homogeneous smooth and hydrophobic surfaces. Cell adhesion was significantly reduced on all silicones compared to the negative control. Proliferation index and cell viability were not influenced much. For development of a new glaucoma drainage device, the silicones Silbione LSR 4330 and Silbione LSR 4350, in this study, with low cell counts for hTF and low proliferation indices for hSF, and silicone Silastic MDX4-4210, with low cell counts for hSF and low proliferation indices for hTF, have shown the best results in vitro. Due to the high cell adhesion shown on Silicone LSR 40, 40,026, this material is unsuitable.

Nanosized CaP-silk fibroin-PCL-PEG-PCL/PCL based bilayer membranes for guided bone regeneration

Guided bone regeneration (GBR) concept has been developed to prevent the formation of non-functional scar tissue layer on defect site by undertaking barrier role. In this study, a new bilayer membrane which consisted of one layer of electrospun silk fibroin/PCL-PEG-PCL incorporating nanocalcium phosphate (SPCA)11Silk fibroin/PCL-PEG-PCL incorporating nanocalcium phosphate (SPCA) and one layer of PCL membrane was developed for GBR. To improve the osteoconductivity of membranes, nanosized calcium phosphate particles synthesized by Flame Spray Pyrolysis method were incorporated into membranes at 10% (wt) (SPCA10) and 20% (wt) (SPCA20) of the polymer content. The structural and chemical analyses revealed the well-integrated two layers of membranes with a total thickness of ca 100μm. In the regenerative layer, the highly porous mesh structure had a thickness of 12.6μm with randomly oriented fibers having diameters around 760nm, and nanoparticles dispersed homogenously. The mechanical test results showed remarkable improvement on the tensile strength of membranes with incorporation of nanoparticles. Higher water affinity of nanoCaP included membranes was proved by lower contact angle values and higher percent water uptake capacity. Biomineralization assay revealed that nucleation and growth of apatites around fibers of SPCA10 and SPCA20 were apparent while on SPCA0 apatite minerals were barely detected after 10days. Human dental pulp stem cells (DPSC) were seeded on electrospun layer of the bilayer membranes for biocompatibility and osteo-compatibility study. Increasing nanoCaP amount resulted in higher cell adhesion, proliferation, ALP activity and calcium deposition on membranes. These overall results confirmed the biocompatibility and potential applicability of proposed membranes for GBR treatments.

Adhesion and migration of CHO cells on micropatterned single layer graphene

Cell patterning technology on single layer graphene (SLG) is a fairly new field that can find applications in tissue engineering and biomaterial/biosensors development. Recently, we have developed a simple and effective approach for the fabrication of patterned SLG substrates by laser micromachining, and we have successfully applied it for the obtainment of geometrically ordered neural networks. Here, we exploit the same approach to investigate the generalization of the cell response to the surface cues of the fabricated substrates and, contextually, to quantify cell adhesion on the different areas of the patterns. To attain this goal, we tested Chinese hamster ovary (CHO) cells on PDL-coated micropatterned SLG substrates and quantified the adhesion by using single cell force spectroscopy (SCFS). Our results indicate higher cell adhesion on PDL-SLG, and, consequently, an initial CHO cell accumulation on the graphene areas, confirming the neuronal behaviour observed previously; interestingly, at later time point in culture, cell migration was observed towards the adjacent SLG ablated regions, which resulted more favourable for cell proliferation. Therefore, our findings indicate that the mechanism of interaction with the surface cues offered by the micropatterned substrates is strictly cell-type dependent.

Preparation of elastic chitosan/poly(l-lactide-co-ε-caprolactone) macroporous scaffolds by acetylation and particulate leaching

For soft tissue engineering applications, 3-D macroporous acetylated chitosan/poly(l-lactideco-e-caprolactone) (PLCL) scaffolds were prepared by acetylation and particulate leaching using sodium acetate in an acidic water/dioxane solution. Acetylated 5 wt% chitosan/PLCL scaffold of 90% porosity was determined and confirmed through various tests. The physiochemical properties of acetylated chitosan/PLCL hybrid scaffolds were examined by measuring water contact angles, pore morphology and interconnectivity using scanning electron microscopy (SEM), and dye release testing. In addition, mechanical properties such as tensile strength and bending stress recovery for determining the elasticity of scaffolds were measured. The fibroblast cell line NIH-3T3 was used to test relative cell affinities for the acetylated chitosan/PLCL vs. normal chitosan/PLCL films and porous scaffolds. The acetylated chitosan/PLCL films and scaffolds showed a high initial cell adhesion after 4 h of cell culture and increased cell proliferation compared to that of the control. The acetylated chitosan/PLCL scaffolds produced by particulate leaching showed a highly porous structure and improved the biocompatibility and stability of chitosan compared to that of chitosan-coated PLCL scaffolds. Thus, these scaffolds may be very useful for a variety of tissue engineering applications.

Degradation and biocompatibility of photoembossed PLGA-acrylate blend for improved cell adhesion.

We have shown previously that PMMA-acrylate photopolymers are biocomopatible and can exhibit improved cell adhesion compared to PMMA, due to an increase in negative surface charge caused by UV radiation PLGA has been used widely in soft tissue regeneration due to its high biocompatibility and cell adhesion. This polymer is also biodegradable and can be utilised in the field of vascular regeneration. In this study, PLGA is blended with a triacrylate monomer (TPETA) to create a degradable photopolymer blend. Surface relief structures are formed on this PLGA-TPETA by photoembossing. An optimum height of 950 nm was achieved for a 10 µm pitch with the height of these relief structures being controlled by changing UV intensity, processing temperature and time. Degradation studies of this blend revealed a bulk degradation mechanism with PLGA-TPETA degrading slower compared to pure PLGA. We also evaluated the adhesion of human umbilical vein endothelial cells (HUVECs) on both smooth and textured PLGA-TPETA films. Embossed PLGA-TPETA films showed improved cell adhesion compared to smooth substrates. Furthermore, HUVECs proliferated faster on the embossed surface compared to their smooth counterparts. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 163-171, 2018.

Effect of Hydrofluoric Acid Etching Time on Titanium Topography, Chemistry, Wettability, and Cell Adhesion.

Titanium implant surface etching has proven an effective method to enhance cell attachment. Despite the frequent use of hydrofluoric (HF) acid, many questions remain unresolved, including the optimal etching time and its effect on surface and biological properties. The objective of this study was to investigate the effect of HF acid etching time on Ti topography, surface chemistry, wettability, and cell adhesion. These data are useful to design improved acid treatment and obtain an improved cell response. The surface topography, chemistry, dynamic wetting, and cell adhesiveness of polished Ti surfaces were evaluated after treatment with HF acid solution for 0, 2; 3, 5, 7, or 10 min, revealing a time-dependent effect of HF acid on their topography, chemistry, and wetting. Roughness and wetting increased with longer etching time except at 10 min, when roughness increased but wetness decreased. Skewness became negative after etching and kurtosis tended to 3 with longer etching time. Highest cell adhesion was achieved after 5–7 min of etching time. Wetting and cell adhesion were reduced on the highly rough surfaces obtained after 10-min etching time.

Novel bioceramic-reinforced hydrogel for alveolar bone regeneration.

The osseointegration of dental implants and their consequent long-term success is guaranteed by the presence, in the extraction site, of healthy and sufficient alveolar bone. Bone deficiencies may be the result of extraction traumas, periodontal disease and infection. In these cases, placement of titanium implants is contraindicated until a vertical bone augmentation is obtained. This goal is achieved using bone graft materials, which should simulate extracellular matrix (ECM), in order to promote osteoblast proliferation and fill the void, maintaining the space without collapsing until the new bone is formed. In this work, we design, develop and characterize a novel, moldable chitosan-pectin hydrogel reinforced by biphasic calcium phosphate particles with size in the range of 100-300μm. The polysaccharide nature of the hydrogel mimics the ECM of natural bone, and the ceramic particles promote high osteoblast proliferation, assessed by Scanning Electron Microscopy analysis. Swelling properties allow significant adsorption of water solution (up to 200% of solution content) so that the bone defect space can be filled by the material in an in vivo scenario. The incorporation of ceramic particles makes the material stable at different pH and increases the compressive elastic modulus, toughness and ultimate tensile strength. Furthermore, cell studies with SAOS-2 human osteoblastic cell line show high cell proliferation and adhesion already after 72h, and the presence of ceramic particles increases the expression of alkaline phosphatase activity after 1week. These results suggest a great potential of the developed moldable biomaterials for the regeneration of the alveolar bone. The positive fate of a surgical procedure involving the insertion of a titanium screw still depends on the quality and quantity of alveolar bone which is present in the extraction site. Available materials are basically hard scaffold materials with un-predictable behavior in different condition and difficult shaping properties. In this work we developed a novel pectin-chitosan hydrogel reinforced with ceramic particles. Polysaccharides simulate the extracellular matrix of natural bone and the extensive in vitro cells culture study allows to assess that the incorporation of the ceramic particles promote a pro-osteogenic response. Shape control, easy adaption of the extraction site, predictable behavior in different environment condition, swelling properties and an anti-inflammatory response are the significant characteristics of the developed biomaterial.

Fabrication of silk fibroin based three dimensional scaffolds for tissue engineering

Herein we report successful synthesis of silk fibroin (SF) three dimensional scaffolds (SF 3D-scaffold) from SF sponge and SF nanofibers. Both the nanofibers and sponge were prepared from Bombyx mori fibroin. The SF 3D-scaffold was prepared by electrospinning the fibroin nanofibers over the sponge. Surface morphology was determined by scanning electron microscopy (SEM), while nanofiber diameter and pore size were measured using imageJ software. Effect of spinning time on the pore size and cell adhesion was determined. Average diameter of the SF nanofibers was measured to be 320 nm and pore size was found to reduce with increasing spinning time, such that, for 1 h spinning time pore size was 231 µm and the same for 3.5 h was 4.1 µm. However, the number of pores increased with spinning time. The results confirmed adhesion of MC3T3-E1 cells on the SF sponge, SF nanofibers and SF three dimensional scaffolds. Higher cell adhesion was found on the three dimensional scaffold in comparison to the nanofibers and sponge, possibly due to highly porous structure with very small and numerous pores in the resultant composite; hence more cell adhesion sites. The cell adhesion result confirmed biocompatibility of the SF 3D-scaffold and hence its suitability for applications in tissue engineering.

Enhanced cell adhesion on bioinert ceramics mediated by the osteogenic cell membrane enzyme alkaline phosphatase

Functional bone and dental implant materials are required to guide cell response, offering cues that provide specific instructions to cells at the implant/tissue interface while maintaining full biocompatibility as well as the desired structural requirements and functions. In this work we investigate the influence of covalently immobilized alkaline phosphatase (ALP), an enzyme involved in bone mineralization, on the first contact and initial cell adhesion. To this end, ALP is covalently immobilized by carbodiimide-mediated chemoligation on two highly bioinert ceramics, alpha-alumina (Al2O3) and yttria-stabilized zirconia (Y-TZP) that are well-established for load-bearing applications. The physicochemical surface properties are evaluated by profilometry, zeta potential and water contact angle measurements. The initial cell adhesion of human osteoblasts (HOBs), human osteoblast-like cells (MG-63) and mesenchymal stromal cells (hMSCs) was investigated. Cell adhesion was assessed at serum free condition via quantification of percentage of adherent cells, adhesion area and staining of the focal adhesion protein vinculin. Our findings show that after ALP immobilization, the Al2O3 and Y-TZP surfaces gained a negative charge and their hydrophilicity was increased. In the presence of surface-immobilized ALP, a higher cell adhesion, more pronounced cell spreading and a higher number of focal contact points were found. Thereby, this work gives evidence that surface functionalization with ALP can be utilized to modify inert materials for biological conversion and faster bone regeneration on inert and potentially load-bearing implant materials.

Macrophages adhesion rate on Ti-6Al-4V substrates: polishing and DLC coating effects

Introduction Various works have shown that diamond-like carbon (DLC) coatings are able to improve the cells adhesion on prosthesis material and also cause protection against the physical wear. On the other hand there are reports about the effect of substrate polishing, in evidence of that roughness can enhance cell adhesion. In order to compare and quantify the joint effects of both factors, i.e, polishing and DLC coating, a commonly prosthesis material, the Ti-6Al-4V alloy, was used as raw material for substrates in our studies of macrophage cell adhesion rate on rough and polished samples, coated and uncoated with DLC. Methods The films were produced by PECVD technique on Ti-6Al-4V substrates and characterized by optical profilometry, scanning electron microscopy and Raman spectroscopy. The amount of cells was measured by particle analysis in IMAGE J software. Cytotoxicity tests were also carried out to infer the biocompatibility of the samples. Results The results showed that higher the surface roughness of the alloy, higher are the cells fixing on the samples surface, moreover group of samples with DLC favored the cell adhesion more than their respective uncoated groups. The cytotoxity tests confirmed that all samples were biocompatible independently of being polished or coated with DLC. Conclusion From the observed results, it was found that the rougher substrate coated with DLC showed a higher cell adhesion than the polished samples, either coated or uncoated with the film. It is concluded that the roughness of the Ti-6Al-4V alloy and the DLC coating act complementary to enhance cell adhesion.

Surface biofunctionalization of β-TCP blocks using aptamer 74 for bone tissue engineering

Successful bone regeneration following oral and maxillofacial surgeries depends on efficient functionalization strategies that allow the recruitment of osteogenic progenitor cells at the tissue/implant interface. We have previously identified aptamer 74, which exhibited a binding affinity for osteogenically induced jaw periosteal cells (JPCs). In the present study, this aptamer was used for the surface biofunctionalization of β-tricalcium phosphate (β-TCP) blocks.Atomic force microscopy (AFM) measurements showed increased binding activity of aptamer 74 towards osteogenically induced JPCs compared to untreated controls. The immobilization efficiency of aptamer 74 was analyzed using the QuantiFluor ssDNA assay for 2D surfaces and by amino acid analysis for 3D β-TCP constructs. Following the successful immobilization of aptamer 74 in 2D culture wells and on 3D constructs, in vitro assays showed no significant differences in cell proliferation compared to unmodified surfaces. Interestingly, JPC mineralization was significantly higher on the 2D surfaces and higher cell adhesion was detected on the 3D constructs with immobilized aptamer.Herein, we report an established, biocompatible β-TCP matrix with surface immobilization of aptamer 74, which enhances properties such as cell adhesion on 3D constructs and mineralization on 2D surfaces. Further studies need to be performed to improve the immobilization efficiency and to develop a suitable approach for JPC mineralization growing within 3D β-TCP constructs.

Cell adhesion induces overexpression of chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) and contributes to cell adhesion-mediated drug resistance (CAM-DR) in multiple myeloma cells

Previous studies have shown that chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) exerts its anti-apoptotic function in many solid cancers. However, its role in human multiple myeloma (MM) has not been thoroughly elucidated. In this study, we investigate the role of CHD1L in MM. Preliminarily, up-regulation and down-regulation assay verified that CHD1L exerts its anti-apoptotic role through the apoptotic pathway involving caspase-9–caspase-3 apoptotic pathway in MM cells. In addition, we determined that CHD1L expression is increased when MM cells were adhered to fibronectin (FN) or bone marrow stromal cell line HS-5 cells and cell adhesion assay indicated that CHD1L siRNA reversed the high cell adhesion rate. Consistent with the reduced adhesion rate, the cells translated to a compromised cell adhesion-mediated drug resistance (CAM-DR) phenotype in MM. In summary, we will propose strategies for developing a CHD1L inhibitor for potential treatment of MM.

Mussel-Inspired Anchoring of Polymer Loops That Provide Superior Surface Lubrication and Antifouling Properties.

We describe robustly anchored triblock copolymers that adopt loop conformations on surfaces and endow them with unprecedented lubricating and antifouling properties. The triblocks have two end blocks with catechol-anchoring groups and a looping poly(ethylene oxide) (PEO) midblock. The loops mediate strong steric repulsion between two mica surfaces. When sheared at constant speeds of ∼2.5 μm/s, the surfaces exhibit an extremely low friction coefficient of ∼0.002-0.004 without any signs of damage up to pressures of ∼2-3 MPa that are close to most biological bearing systems. Moreover, the polymer loops enhance inhibition of cell adhesion and proliferation compared to polymers in the random coil or brush conformations. These results demonstrate that strongly anchored polymer loops are effective for high lubrication and low cell adhesion and represent a promising candidate for the development of specialized high-performance biomedical coatings.

Effect of hydroxyapatite materials properties on fibronectin-mediated breast cancer cell adhesion and pro-angiogenic secretion

Event Abstract Back to Event Effect of hydroxyapatite materials properties on fibronectin-mediated breast cancer cell adhesion and pro-angiogenic secretion Fei Wu1, Weisi Chen2, Brian Gillis1, Claudia Fischbach2, 3, Lara Estroff1, 3 and Delphine Gourdon1, 2 1 Cornell University, Materials Science and Engineering, United States 2 Cornell University, Biomedical Engineering, United States 3 Kavli Institute at Cornell for Nanoscale Science, United States Introduction: Breast cancer preferentially metastasizes to bone and induces pathological remodeling. There has been evidence suggesting that the materials properties of bone mineral may play an important role in breast cancer metastasis[1],[2]. Hydroxyapatite (HAP, Ca10(PO4)6(OH)2) is a calcium phosphate mineral with structural and mechanical properties closely related to bone apatite. In this study, we seek to understand: (i) whether HAP materials properties, such as surface chemistry and roughness, alter the mineral/extracellular matrix (ECM) interface in bone, and (ii) how the altered ECM impacts breast cancer cell functions. Specifically, we investigated the conformation of fibronectin (Fn, a major ECM protein), breast cancer cell adhesion and pro-angiogenic secretion as a function of underlying HAP materials properties and associated Fn conformation. Materials and Methods: Geologic HAP crystals were bought from Etsy. Fibronectin (Fn) was obtained from Life Technologies, NY, and labeled for intramolecular Fӧrster resonance energy transfer (FRET) as previously described[3]. Smooth HAP facets (100)S and (001)S and rough HAP facets (100)R and (001)R were obtained by polishing HAP slabs to roughnesses either below 1 nm, i.e., smaller than Fn radius of gyration, or around 200 nm. Then, we incubated Fn (10% FRET-labeled) on HAP facets and analyzed Fn conformation via FRET. Finally, MDA-MB-231 breast cancer cells (ATCC, VA) were seeded onto Fn coated HAP facets to investigate cell adhesion and secretion of vascular endothelial growth factors (VEGF). Student’s t-tests were used to determine statistical significance between conditions in GraphPad Prism (GraphPad Software Inc., CA). Results and Discussion: We first quantified Fn conformation adsorbed onto HAP via FRET (Figure 1). FRET ratios of Fn adsorbed onto (100) were higher than those onto (001) for both smooth and rough HAP, indicating that Fn adsorbed in more compact conformations onto (100) facets. Interestingly, Fn FRET ratios were not significantly different between smooth and rough HAP, suggesting that Fn conformation was independent of roughness within the range of interest. We then assessed MDA-MB-231 cells behavior. Our preliminary results showed (i) higher cell adhesion onto (001) than onto (100) HAP facets, (ii) overall higher cell adhesion onto rough HAP than onto smooth HAP, and (iii) lower VEGF secretion associated with higher adhesion. Conclusions: Our FRET analysis indicated that Fn conformation (% of unfolded Fn) depends exclusively on HAP surface chemistry. In contrast, our cells study revealed enhanced adhesion and decreased pro-angiogenic secretion of cancer cells linked with both (001) HAP facets (coated with more unfolded Fn) and rougher HAP. These findings deconvolute the roles of mineral surface chemistry and roughness in interfacial ECM deposition and have important implications for our understanding of breast cancer bone metastasis. Karin Wang; Jin Ho Chang; Young Hye Song; Siyoung Choi; Maura Weathers

Association of Cell-adhesion Activities with Virulence in Shiga toxin-producing Escherichia coli O103：H2.

The characteristics of 11 strains of Stx1-producing and Stx2-non-producing STEC O103：H2 were analyzed to investigate the differences in virulence in a single serotype of Shiga toxin (Stx) -producing Escherichia coli (STEC). Differences in the cell-adhesion activity to Caco-2 cells were observed among the strains. The activity of the one strain, isolated from a patient with hemolytic uremic syndrome was 4-20-fold higher than those of the other strains. Although the strains with high cell-adhesion activity showed high expressions of eae, espB, espD, and tir in the locus of enterocyte effacement related with cell-adhesion, those were not specific for this strain. In addition, the Stx1 production level of the strain was not particularly high. It was indicated that the high adhesion activity might be a potential factor to associate serious symptom.

Collagen Promotes Higher Adhesion, Survival and Proliferation of Mesenchymal Stem Cells

Mesenchymal stem cells (MSC) can differentiate into several cell types and are desirable candidates for cell therapy and tissue engineering. However, due to poor cell survival, proliferation and differentiation in the patient, the therapy outcomes have not been satisfactory. Although several studies have been done to understand the conditions that promote proliferation, differentiation and migration of MSC in vitro and in vivo, still there is no clear understanding on the effect of non-cellular bio molecules. Of the many factors that influence the cell behavior, the immediate cell microenvironment plays a major role. In this context, we studied the effect of extracellular matrix (ECM) proteins in controlling cell survival, proliferation, migration and directed MSC differentiation. We found that collagen promoted cell proliferation, cell survival under stress and promoted high cell adhesion to the cell culture surface. Increased osteogenic differentiation accompanied by high active RHOA (Ras homology gene family member A) levels was exhibited by MSC cultured on collagen. In conclusion, our study shows that collagen will be a suitable matrix for large scale production of MSC with high survival rate and to obtain high osteogenic differentiation for therapy.

Screening for potential novel probiotic Lactobacillus strains based on high dipeptidyl peptidase IV and α-glucosidase inhibitory activity

Lactobacillus strains were screened for inhibition of dipeptidyl peptidase IV (DPP-IV) and α-glucosidase. Cell-free excretory supernatants (CFS) and cell-free extracts (CFE) of 21 strains were prepared, and most of them showed DPP-IV inhibition in a concentration-dependent manner, which started at the exponential phase and peaked at the stationary phase of the Lactobacillus. The CFS bioactivity was heat-resistant, stable to acid–alkali and glucosidase treatment. Trypsin treatment specifically elevated CFS's DPP-IV inhibition. Lactobacillus plantarum strains CFS displayed different protein patterns and trypsin sensitive bands on SDS-PAGE, demonstrating specific peptide-cleavage contributes to higher inhibition. Seven strains with highest DPP-IV inhibition also displayed α-glucosidase inhibitory activity, of which, three strains of L. plantarum and one of Lactobacillus brevis showed tolerance to simulated gastrointestinal tract conditions and high HT-29 cell adhesion. This initial report of lactobacilli secreting strong DPP-IV and α-glucosidase inhibition components will be beneficial for selection and application of anti-diabetic probiotics.

Probiotic properties of lactic acid bacteria isolated from Korean rice wine Makgeolli

Makgeolli, a traditional Korean alcoholic beverage, includes many kinds of lactic acid bacteria (LAB). In this study, 17 different LAB strains were isolated from makgeolli and their functional effects were examined. The 17 LAB were identified as eight Pediococcus acidilactici strains, six P. pentosaceus strains, two Lactobacillus curvatus strains, and one L. curstorum strain. Evaluation of nitric oxide (NO) production showed that all of the LAB strains except P. acidilactici 4 (PA4) produced higher amounts of NO than L. rhamnosus GG, indicating that they may have immunomodulatory effects. In addition, P. acidilactici 5 ( PA5), P. pentosaceus 5 (PP5), L. curvatus 1, and L. curstorum 1 exhibited high acid tolerance properties. PA5 and PP5 also exhibited good bile acid tolerance. Lastly, Caco-2 cell adhesion experiments revealed that PA5 had a high cell adhesion rate of 19-43%. In conclusion, PA5 may be useful as a probiotic for further food industry applications.

Homogeneous synthesis of amino‐reserved chitosan‐graft‐polycaprolactone in an ionic liquid and the application in cell cultivation

AbstractThe homogeneous preparation of amino‐reserved chitosan‐graft‐polycaprolactone copolymer (ACS‐g‐PCL) was achieved in 1‐butyl‐3‐methylimidazolium acetate via a protection − ring‐opening graft polymerization − deprotection procedure. The molar substitution of PCL (MSPCL) in ACS‐g‐PCL copolymers was in the range 17.1 − 45.6 and could be well controlled by altering reaction conditions. The resultant ACS‐g‐PCL copolymers were soluble in some common organic solvents such as dimethylsulfoxide, ethanol and toluene. As the increment of MSPCL increased, the hydrophilicity of the ACS‐g‐PCL copolymers decreased. Eventually, ACS‐g‐PCL microspheres with a diameter of 100–200 µm were fabricated by an emulsion evaporation method. Among them, the ACS‐g‐PCL copolymer with an MSPCL of 26.8 formed into porous microspheres. The fluorescence microscopy and cytotoxicity results suggested that all of the microspheres, especially the ACS‐g‐PCL microspheres with MSPCL of 17.1 and 26.8, showed high cell adhesion and cytocompatibility to the human osteosarcoma cell line (MG‐63), indicating their great prospects in tissue engineering. © 2015 Society of Chemical Industry