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1.
Front Bioeng Biotechnol ; 11: 1165853, 2023.
Article in English | MEDLINE | ID: mdl-37409165

ABSTRACT

Introduction: The functionalization of titanium (Ti) and titanium alloys (Ti6Al4V) implant surfaces via material-specific peptides influence host/biomaterial interaction. The impact of using peptides as molecular linkers between cells and implant material to improve keratinocyte adhesion is reported. Results: The metal binding peptides (MBP-1, MBP-2) SVSVGMKPSPRP and WDPPTLKRPVSP were selected via phage display and combined with laminin-5 or E-cadherin epithelial cell specific peptides (CSP-1, CSP-2) to engineer four metal-cell specific peptides (MCSPs). Single-cell force spectroscopy and cell adhesion experiments were performed to select the most promising candidate. In vivo tests using the dental implant for rats showed that the selected bi functional peptide not only enabled stable cell adhesion on the trans-gingival part of the dental implant but also arrested the unwanted apical migration of epithelial cells. Conclusion: The results demonstrated the outstanding performance of the bioengineered peptide in improving epithelial adhesion to Ti based implants and pointed towards promising new opportunities for applications in clinical practice.

2.
Sci Rep ; 10(1): 9791, 2020 06 17.
Article in English | MEDLINE | ID: mdl-32555227

ABSTRACT

In recent years, fluorescent nanodiamond (fND) particles containing nitrogen-vacancy (NV) centers gained recognition as an attractive probe for nanoscale cellular imaging and quantum sensing. For these applications, precise localization of fNDs inside of a living cell is essential. Here we propose such a method by simultaneous detection of the signal from the NV centers and the spectroscopic Raman signal from the cells to visualize the nucleus of living cells. However, we show that the commonly used Raman cell signal from the fingerprint region is not suitable for organelle imaging in this case. Therefore, we develop a method for nucleus visualization exploiting the region-specific shape of C-H stretching mode and further use k-means cluster analysis to chemically distinguish the vicinity of fNDs. Our technique enables, within a single scan, to detect fNDs, distinguish by chemical localization whether they have been internalized into cell and simultaneously visualize cell nucleus without any labeling or cell-fixation. We show for the first time spectral colocalization of unmodified high-pressure high-temperature fND probes with the cell nucleus. Our methodology can be, in principle, extended to any red- and near-infrared-luminescent cell-probes and is fully compatible with quantum sensing measurements in living cells.


Subject(s)
Cell Nucleus/ultrastructure , Molecular Imaging/methods , Nanodiamonds , Cell Line, Tumor , Cells, Cultured , Cytological Techniques , Dental Pulp/cytology , Dental Pulp/diagnostic imaging , Fluorescent Dyes , Humans , Spectrum Analysis, Raman
3.
Caries Res ; 54(2): 144-153, 2020.
Article in English | MEDLINE | ID: mdl-32107346

ABSTRACT

The development of new diagnostic technologies based on the light scattering and autofluorescence properties of dental tissues is required to improve the diagnostic ability of initial caries lesions earlier than previously done and promoting the potential of treatment without surgical intervention. The aim of this study is to correlate fluorescence-based results provided by multiphoton microscopy (MPM) with confocal Raman microscopy records using phosphate level at 960 cm-1 and the organic matrix at ∼2,931 cm-1 in healthy and demineralized human enamel. Measurements on 14 teeth were made using two incident lights of different wavelengths, released by confocal Raman microscopy and MPM. Raman phosphate peak intensity at 960 cm-1 along with organic to mineral ratio at (2,931/430 cm-1) and nonlinear optical signals (second harmonic generation [SHG] and intrinsic two-photon excited fluorescence [I2PEF]) were recorded from the demineralized and healthy enamel sites. Raman spectral maps showed that the higher the organic/mineral ratio in the demineralized enamel, the lower the intensity of mineral component in the same zone. MPM revealed new optical indicators of carious lesion as shown by the presence of a red-shifted fluorescence peak in the 650- to 750-nm area of the fluorescence spectrum of demineralized enamel. Moreover, on sample regions with insignificant autofluorescence, the emergence of the SHG signal could be noted. By comparing I2PEF images with the structural motifs observed by the confocal Raman imaging system, the morphological similarity of the acquired images was quite evident. Any change in the I2PEF spectra reflects alterations in the chemical composition of enamel. These findings may provide an important basis for potentially valuable applications of photonic tools in the clinical diagnosis of tooth pathological conditions, besides exposing the fundamental role of organic matrix in enamel integrity and reparation.


Subject(s)
Dental Caries , Tooth , Dental Caries/diagnostic imaging , Dental Enamel/diagnostic imaging , Humans , Phosphates , Tooth Demineralization/diagnostic imaging
4.
ACS Appl Bio Mater ; 3(11): 7531-7539, 2020 Nov 16.
Article in English | MEDLINE | ID: mdl-35019494

ABSTRACT

Biological activities of cells such as survival and differentiation processes are mainly maintained by a specific extracellular matrix (ECM). Hydrogels have recently been employed successfully in tissue engineering applications. In particular, scaffolds made of gelatin methacrylate-based hydrogels (GelMA) showed great potential due to their biocompatibility, biofunctionality, and low mechanical strength. The development of a hydrogel having tunable and appropriate mechanical properties as well as chemical and biological cues was the aim of this work. A synthetic and biological hybrid hydrogel was developed to mimic the biological and mechanical properties of native ECM. A combination of gelatin methacrylate and acrylamide (GelMA-AAm)-based hydrogels was studied, and it showed tunable mechanical properties upon changing the polymer concentrations. Different GelMA-AAm samples were prepared and studied by varying the concentrations of GelMA and AAm (AAm2.5% + GelMA3%, AAm5% + GelMA3%, and AAm5% + GelMA5%). The swelling behavior, biodegradability, physicochemical and mechanical properties of GelMA-AAm were also characterized. The results showed a variation of swelling capability and a tunable elasticity ranging from 4.03 to 24.98 kPa depending on polymer concentrations. Moreover, the podocyte cell morphology, cytoskeleton reorganization and differentiation were evaluated as a function of GelMA-AAm mechanical properties. We concluded that the AAm2.5% + GelMA3% hydrogel sample having an elasticity of 4.03 kPa can mimic the native kidney glomerular basement membrane (GBM) elasticity and allow podocyte cell attachment without the functionalization of the gel surface with adhesion proteins compared to synthetic hydrogels (PAAm). This work will further enhance the knowledge of the behavior of podocyte cells to understand their biological properties in both healthy and diseased states.

5.
ACS Appl Mater Interfaces ; 11(36): 32623-32632, 2019 Sep 11.
Article in English | MEDLINE | ID: mdl-31424195

ABSTRACT

Chronic kidney disease is characterized by a gradual decline in renal function that progresses toward end-stage renal disease. Podocytes are highly specialized glomerular epithelial cells which form with the glomerular basement membrane (GBM) and capillary endothelium the glomerular filtration barrier. GBM is an extracellular matrix (ECM) that acts as a mechanical support and provides biophysical signals that control normal podocytes behavior in the process of glomerular filtration. Thus, the ECM stiffness represents an essential characteristic that controls podocyte function. Hydrolyzed Polyacrylamide (PAAm) hydrogels are smart polyelectrolyte materials. Their biophysical properties can be tuned as desired to mimic the natural ECM. Therefore, these hydrogels are investigated as new ECM-like constructs to engineer a podocyte-like basement membrane that forms with cultured human podocytes a functional glomerular-like filtration barrier. Such ECM-like PAAm hydrogel construct will provide unique opportunity to reveal podocyte cell biological responses in an in vivo-like setting by controlling the physical properties of the PAAm membranes. In this work, Hydrolyzed PAAm scaffolds having different stiffness ranging between 0.6-44 kPa are prepared. The correlation between the hydrogel structural and mechanical properties and Podocyte morphology, elasticity, cytoskeleton reorganization, and podocin expression is evaluated. Results show that hydrolyzed PAAm hydrogels promote good cell adhesion and growth and are suitable materials for the development of future 3D smart scaffolds. In addition, the hydrogel properties can be easily modulated over a wide physiological range by controlling the cross-linker concentration. Finally, tuning the hydrogel properties is an effective strategy to control the cells function. This work addressed the complexity of podocytes behavior which will further enhance our knowledge to develop a kidney-on-chip model much needed in kidney function studies in both healthy and diseased states.


Subject(s)
Acrylic Resins/chemistry , Acrylic Resins/pharmacology , Cell Shape , Hydrogels/chemistry , Hydrogels/pharmacology , Podocytes/cytology , Biomechanical Phenomena , Calorimetry, Differential Scanning , Cell Line , Cell Shape/drug effects , Elastic Modulus , Elasticity , Humans , Hydrolysis , Phenotype , Podocytes/drug effects , Tissue Scaffolds/chemistry
6.
Colloids Surf B Biointerfaces ; 181: 489-497, 2019 Sep 01.
Article in English | MEDLINE | ID: mdl-31176121

ABSTRACT

Extensive use of porous silicon (PSi) for tissue engineering is due to its convenient properties as it is both nontoxic and bioresorbable. Moreover, PSi surface modification is an important step to enhance cell adhesion and proliferation. In this work, a combination of optical and electrochemical studies is performed to elaborate a suitable PSi multilayer substrate for cell culture. For this study, we modified PSi surface by silanization and antibody grafting (APTES-anti STRO1), the 12-mer specific peptide to silicon p + type coating and the peptide modified with the antibody recognition sequence. Electrochemical characterization of PSi multilayers is performed to investigate its electrical behavior, determine the optimal measuring conditions and reveal the most stable PSi surfaces. Then, the behavior of dental pulp stem cells (DPSC) was investigated on various modified PSi surfaces. An electrochemical method was applied for the first time monitoring the electrical behavior of stem cell adhesion. The cells electrochemical behavior depends on the nature of the surface coating and the peptide-anti STRO1 improved adhesion and cell spreading onto the PSi surface compared to bare surface and the one coated with the peptide. Fluorescent microscopy revealed that all surface modification methods enhance cell adhesion compared to the bare PSi surface. An increased cell number is observed on APTES-anti STRO1, peptide and peptide-anti STRO1 coated PSi. The peptide-anti STRO1 provided the best cell proliferation results suggesting the improved accessibility of the recognition fragment of the antibody anti-STRO1.


Subject(s)
Dental Pulp/cytology , Electrochemical Techniques , Optical Imaging , Silicon/chemistry , Stem Cells/cytology , Cell Adhesion , Cell Proliferation , Cells, Cultured , Humans , Particle Size , Porosity , Surface Properties
7.
Sci Rep ; 8(1): 5917, 2018 04 12.
Article in English | MEDLINE | ID: mdl-29650983

ABSTRACT

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that affects the motor system leading to generalized paralysis and death of patients. The understanding of early pathogenic mechanisms will help to define early diagnostics criteria that will eventually provide basis for efficient therapeutics. Early symptoms of ALS usually include muscle weakness or stiffness. Therefore, mechanical response of differentiated myotubes from primary cultures of mice, expressing the ALS-causing SOD1 G93A mutation, was examined by atomic force microscopy. Simultaneous acquisition of topography and cell elasticity of ALS myotubes was performed by force mapping method, compared with healthy myotubes and supplemented with immunofluorescence and qRT-PCR studies. Wild type myotubes reveal a significant difference in elasticity between a narrow and a wide population, consistent with maturation occurring with higher actin expression relative to myosin together with larger myotube width. However, this is not true for SOD1 G93A expressing myotubes, where a significant shift of thin population towards higher elastic modulus values was observed. We provide evidence that SOD1 mutant induces structural changes that occurs very early in muscle development and well before symptomatic stage of the disease. These findings could significantly contribute to the understanding of the role of skeletal muscle in ALS pathogenesis.


Subject(s)
Amyotrophic Lateral Sclerosis/genetics , Muscle Development/genetics , Muscle Fibers, Skeletal/chemistry , Superoxide Dismutase-1/genetics , Actins/genetics , Amyotrophic Lateral Sclerosis/physiopathology , Animals , Cell Differentiation/genetics , Disease Models, Animal , Elasticity/physiology , Gene Expression Regulation/drug effects , Humans , Mechanical Phenomena , Mice , Microscopy, Atomic Force , Muscle Fibers, Skeletal/pathology , Muscle Fibers, Skeletal/ultrastructure , Muscle Weakness/genetics , Mutation , Myosins/genetics , Superoxide Dismutase-1/chemistry
8.
J Biomed Opt ; 22(8): 1-8, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28822139

ABSTRACT

The separation zone between enamel and dentin [dentin-enamel junction (DEJ)] with different properties in biomechanical composition has an important role in preventing crack propagation from enamel to dentin. The understanding of the chemical structure (inorganic and organic components), physical properties, and chemical composition of the human DEJ could benefit biomimetic materials in dentistry. Spatial distribution of calcium phosphate crystallinity and the collagen crosslinks near DEJ were studied using confocal Raman microscopy and calculated by different methods. To obtain collagen crosslinking, the ratio of two peaks 1660 cm-1 over 1690 cm-1 (amide I bands) is calculated. For crystallinity, the inverse full-width at half maximum of phosphate peak at 960 cm-1, and the ratio of two Raman peaks of phosphate at 960/950 cm-1 is provided. In conclusion, the study of chemical and physical properties of DEJ provides many benefits in the biomaterial field to improve the synthesis of dental materials in respect to the natural properties of human teeth. Confocal Raman microscopy as a powerful tool provides the molecular structure to identify the changes along DEJ and can be expanded for other mineralized tissues.


Subject(s)
Collagen/ultrastructure , Dental Enamel/ultrastructure , Dentin/ultrastructure , Microscopy, Confocal/methods , Nonlinear Optical Microscopy/methods , Humans , Tooth/ultrastructure
9.
J Biomed Opt ; 20(7): 076013, 2015 Jul.
Article in English | MEDLINE | ID: mdl-26216272

ABSTRACT

Regenerative medicine brings promising applications for mesenchymal stem cells, such as dental pulp stem cells (DPSCs). Confocal Raman microscopy, a noninvasive technique, is used to study osteogenic differentiation of DPSCs. Integrated Raman intensities in the 2800 to 3000 cm⁻¹ region (C-H stretching) and the 960 cm⁻¹ peak (ν1 PO4³â») were collected (to image cells and phosphate, respectively), and the ratio of two peaks 1660 over 1690 cm⁻¹ (amide I bands) to measure the collagen cross-linking has been calculated. Raman spectra of DPSCs after 21 days differentiation reveal several phosphate peaks: ν1 (first stretching mode) at 960 cm⁻¹, ν2 at 430 cm⁻¹, and ν4 at 585 cm⁻¹ and collagen cross-linking can also be calculated. Confocal Raman microscopy enables monitoring osteogenic differentiation in vitro and can be a credible tool for clinical stem cell based research.


Subject(s)
Cell Differentiation/physiology , Dental Pulp/cytology , Extracellular Matrix/chemistry , Microscopy, Confocal/methods , Spectrum Analysis, Raman/methods , Stem Cells/chemistry , Adolescent , Collagen , Humans , Stem Cells/cytology
10.
Macromol Biosci ; 14(12): 1771-82, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25212873

ABSTRACT

Further development of biomaterials is expected as advanced therapeutic products must be compliant to good manufacturing practice regulations. A spraying method for building-up polyelectrolyte films followed by the deposition of dental pulp cells by spraying is presented. Physical treatments of UV irradiation and a drying/wetting process are applied to the system. Structural changes and elasticity modifications of the obtained coatings are revealed by atomic force microscopy and by Raman spectroscopy. This procedure results in thicker, rougher and stiffer film. The initially ordered structure composed of mainly α helices is transformed into random/ß-structures. The treatment enhanced dental pulp cell adhesion and proliferation, suggesting that this system is promising for medical applications.


Subject(s)
Biocompatible Materials/chemistry , Dental Pulp/metabolism , Membranes, Artificial , Polyglutamic Acid/chemistry , Polylysine/chemistry , Adolescent , Cell Survival , Cells, Cultured , Dental Pulp/cytology , Female , Humans , Male , Ultraviolet Rays , Wettability
11.
J Chem Inf Model ; 53(12): 3273-9, 2013 Dec 23.
Article in English | MEDLINE | ID: mdl-24289530

ABSTRACT

Engineering shape-controlled bionanomaterials requires comprehensive understanding of interactions between biomolecules and inorganic surfaces. We explore the origin of facet-selective binding of peptides adsorbed onto Pt(100) and Pt(111) crystallographic planes. Using molecular dynamics simulations, we show that upon adsorption the peptides adopt a predictable conformation. We compute the binding energies of the amino acids constituting two adhesion peptides for Pt, S7, and T7 and demonstrate that peptides' surface recognition behavior that makes them unique among populations originates from differential adsorption of their building blocks. We find that the degree of peptide binding is mainly due to polar amino acids and the molecular architecture of the peptides close to the Pt facets. Our analysis is a first step in the prediction of enhanced affinity between inorganic materials and a peptides, toward the synthesis of novel nanomaterials with programmable shape, structure, and properties.


Subject(s)
Amino Acids/chemistry , Peptides/chemistry , Platinum/chemistry , Adsorption , Crystallization , Molecular Dynamics Simulation , Nanostructures , Protein Binding , Protein Conformation , Surface Properties , Thermodynamics
12.
J Biophotonics ; 6(4): 330-7, 2013 Apr.
Article in English | MEDLINE | ID: mdl-22815209

ABSTRACT

Multiphoton microscopy has been used to reveal structural details of dentine and enamel at the dentin-enamel junction (DEJ) based on their 2-photon excited fluorescence (2PEF) emission and second harmonic generation (SHG). In dentine tubule 2PEF intensity varies due to protein content variation. Intertubular dentin produces both SHG and 2PEF signals. Tubules are surrounded by a thin circular zone with a lower SHG signal than the bulk dentine and the presence of collagen fibers perpendicular to the tubule longitudinal axis is indicated by strong SHG responses. The DEJ appears as a low intensity line on the 2PEF images and this was never previously reported. The SHG signal is completely absent for enamel and aprismatic enamel shows a homogeneous low 2PEF signal contrary to prismatic enamel. The SHG intensity of mantle dentine is increasing from the dentine-enamel junction in the first 12 µm indicating a progressive presence of fibrillar collagen and corresponding to the more external part of mantle dentine where matrix metallo-proteases accumulate. The high information content of multiphoton images confirms the huge potential of this method to investigate tooth structures in physiological and pathological conditions.


Subject(s)
Dental Enamel/cytology , Dentin/cytology , Microscopy, Fluorescence, Multiphoton/methods , Calcium Phosphates/metabolism , Collagen/metabolism , Dentin/metabolism , Humans
13.
Clin Oral Investig ; 17(3): 757-63, 2013 Apr.
Article in English | MEDLINE | ID: mdl-22855265

ABSTRACT

OBJECTIVES: Our aim was to determine the origin of the red fluorescence of carious dentine observed with the Soprolife® camera. METHODS: We conducted in vitro studies to evaluate the origin of the red fluorescence using acids and matrix metalloproteinase (MMP) to mimic caries and methylglycoxal (MGO) to evaluate the effect of glycation reactions on the red fluorescence. In every step of these models, we detected the changes of dentin photonic response with Soprolife® in daylight mode and in treatment mode. A Raman spectroscopy analysis was performed to determine the variations of the dentin organic during the in vitro caries processes. Raman microscopy was performed to identify change in the collagen matrix of dentine. RESULTS: The red fluorescence observed in carious dentine using a Soprolife® camera corresponds to the brownish color observed using daylight. Demineralization using nitric acid induces a loss of the green fluorescence of dentine. The red fluorescence of carious dentine is resistant to acid treatment. Immersion of demineralized dentine in MGO induces a change of color from white to orange-red. This indicates that the Maillard reaction contributes to lesion coloration. Immersion of demineralized dentine in an MMP-1 solution followed by MGO treatment results in a similar red fluorescence. Raman microspectroscopy analysis reveals accumulation of AGE's product in red-colored dentine. CONCLUSIONS: Our results provide important information on the origin of the fluorescence variation of dentine observed with the Soprolife® camera. We demonstrate that the red fluorescence of carious dentine is linked to the accumulation of Advanced Glycation End products (AGE). CLINICAL RELEVANCE: The study provides a new biological basis for the red fluorescence of carious dentine and reinforces the importance of the Soprolife® camera in caries diagnostics.


Subject(s)
Dental Caries/pathology , Dentin/pathology , Photography, Dental , Collagen/chemistry , Dental Caries/diagnosis , Dentin/chemistry , Fluorescence , Glycation End Products, Advanced/analysis , Humans , Maillard Reaction , Matrix Metalloproteinase 1 , Photography, Dental/instrumentation , Spectrum Analysis, Raman
14.
Biomed Tech (Berl) ; 57(6): 481-9, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23183721

ABSTRACT

BACKGROUND: A combinatorial phage display approach was previously used to evolve a 12-mer peptide (SVSVGMKPSPRP) with the highest affinity for different semiconductor surfaces. The discovery of the multiple occurrences of the SVSVGMKPSPRP sequence in an all-against-all basic local alignment search tool search of PepBank sequences was unexpected, and a Google search using the peptide sequence recovered 58 results concerning 12 patents and 16 scientific publications. The number of patent and articles indicates that the peptide is perhaps a broad range adhesion peptide. METHODS: To evaluate peptide properties, we conducted a study to investigate peptide adhesion on different inorganic substrates by mass spectrometry and atomic force microscopy for gold, carbon nanotubes, cobalt, chrome alloy, titanium, and titanium alloy substrates. RESULTS: Our results showed that the peptide has a great potential as a linker to functionalize metallic surfaces if specificity is not a key factor. This peptide is not specific to a particular metal surface, but it is a good linker for the functionalization of a wide range of metallic materials. CONCLUSION: The fact that this peptide has the potential to adsorb on a large set of inorganic surfaces suggests novel promising directions for further investigation. Affinity determination of SVSVGMKPSPRP peptide would be an important issue for eventual commercial uses.


Subject(s)
Biocompatible Materials/chemistry , Cell Adhesion Molecules/chemistry , Metals/chemistry , Peptides/chemistry , Materials Testing , Protein Binding
15.
J Mater Sci Mater Med ; 23(6): 1343-53, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22447122

ABSTRACT

The aim of this study was to evaluate the changes in the transition layer at the interface between yttria partial stabilized tetragonal zirconia polycrystalline (Y-TZP) core and veneering feldspathic ceramic (VITA VM(®)9), under different manufacturing methods. Confocal Raman microscopy and energy dispersive X-ray spectroscopy (EDS) analyses were carried out on tapered veneered cross sections of the interface. For some samples, an additional firing of the core was used, as the application of an optional liner material between the core and veneer. Single Raman spectra were distinguishable between Y-TZP and the veneering materials. VM(®)9 and liner spectra were broadly superimposable. No substantial differences appeared in their chemical elemental composition. 2D Raman images and EDS analysis emphasized changes in the interdiffusion thickness; the additional firing of the core decreased the interdiffusion zone, and the highest firing temperature of the liner increased the interdiffusion zone. These results, which will help us understand the changes in this transition layer, are discussed.


Subject(s)
Ceramics/chemistry , Dental Materials/chemistry , Dental Veneers , Yttrium/chemistry , Zirconium/chemistry , Dental Restoration Failure , Hot Temperature , Humans , Materials Testing , Microscopy, Confocal/methods , Microscopy, Electron, Scanning , Spectrometry, X-Ray Emission , Spectrum Analysis, Raman/methods
16.
Dent Mater ; 28(6): 661-71, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22445442

ABSTRACT

OBJECTIVES: Esthetic demands and biocompatibility have prompted the development of all-ceramic dental crowns. Yttria tetragonal zirconia polycrystalline (Y-TZP) framework material has the best mechanical properties compared to other all-ceramic systems, but the interface is the weakest component of core veneered restorations. Confocal Raman microscopy possibilities are used to ensure the understanding of the zirconia-feldspathic ceramic relationship, which is not well known. METHODS: Bilayered zirconia (Vita In-Ceram(®) YZ) veneer (Vita VM(®)9) blocks were manufactured. Raman analyses were performed using two protocols: (1) single spectra, line scans and images on a sectioned and polished specimen and (2) in depth line scans on unprepared specimen. Single spectra, images and line scans provide information about the crystalline phases, their distribution and the existence of a possible diffusion at the Y-TZP/VM(®)9 interface, respectively. The elemental distribution of zirconium (Zr) and silicon (Si) around this interface were investigated using energy dispersive X-ray spectroscopy (EDS). RESULTS: Raman single spectra embodied a unique spectrum (crystalline) on Y-TZP and two spectra (crystalline and amorphous) on VM(®)9; these spectra were clearly distinguished. Raman line scans showed a series of transition spectra across the interface from VM(®)9 to Y-TZP. It emphasized an interdiffusion zone, which was estimated at a maximum of 2 microns, found on 2d Raman images and confirmed by EDS. The elemental distribution with EDS showed a mutual diffusion of Zr and Si and was mainly dominated by Si diffusion in Y-TZP. SIGNIFICANCE: Confocal Raman microscopy highlights an interdiffusion zone at the zirconia-feldspathic ceramic interface. The elemental transition layer is estimated and is supported by EDS analysis as a coupling technique.


Subject(s)
Ceramics/chemistry , Crowns , Dental Bonding/methods , Dental Veneers , Zirconium/chemistry , Dental Stress Analysis , Materials Testing , Spectrometry, X-Ray Emission , Spectrum Analysis, Raman
17.
J Pept Sci ; 17(2): 143-7, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21234986

ABSTRACT

Considerable advances in materials science are expected via the use of selected or designed peptides to recognize material, control their growth, or to assemble them into elaborate novel devices. Identifying specific peptides for a number of technologically useful materials has been the challenge of many research groups in recent years. This can be accomplished by using affinity-based bio-panning methods such as phage display technologies. In this work, a combinatorial library including billions of clones of genetically engineered M13 bacteriophage was used to select peptides that could recognize improved indium nitride (InN) semiconductor (SC) material. Several rounds of biopanning were necessary to select the phage with the higher affinity from the low variant library. The DNA of this specific phage was extracted and sequenced to set up the related specific adherent peptide. Atomic force microscopy (AFM) is used to demonstrate the real affinity of a selected phage for the InN surface. Due to the possibility of its functionalization with biomolecules and its important physical properties, InN is a promising candidate for developing affinity-based optical and electrical biosensors and/or for biomimetic applications.


Subject(s)
Indium/chemistry , Peptides/chemistry , Semiconductors , Microscopy, Atomic Force , Peptide Library
18.
J Investig Clin Dent ; 2(1): 29-37, 2011 Feb.
Article in English | MEDLINE | ID: mdl-25427325

ABSTRACT

AIM: To evaluate the efficacy of a 2-week administration of a 0.1% chlorhexidine mouthwash in the short-term treatment of chronic periodontitis patients and the impact of this product when administered twice by pocket irrigation. METHODS: Sixty patients were enrolled in a single-centre, placebo-controlled, randomized study with the blind allocation of product to two parallel groups. Clinical assessments were performed, and samples from six selected subgingival sites were collected for microbial analysis by culture at baseline, D15 and D56. Three of the six sites were randomly selected and were treated by subgingival irrigation with the same 0.1% chlorhexidine product at D0 and D7. A subsequent statistical analysis was performed using the paired Student's t-test and Wilcoxon rank sum test for within-group analyses; analysis of variance and the Kruskall-Wallis test were used for between-group analyses. RESULTS: Two-week treatment with a 0.1% chlorhexidine mouthwash slightly reduced the gingival inflammation associated with periodontitis. We observed a significant decrease in Gram-negative, facultative anaerobes and micro-aerophiles, and a significant increase in Gram-positive cocci. No increase in the treatment effect was demonstrated by irrigation of the periodontal pockets. CONCLUSION: The 0.1% chlorhexidine mouthwash showed limited beneficial effects in the treatment of periodontitis patients.


Subject(s)
Chlorhexidine/therapeutic use , Chronic Periodontitis/drug therapy , Mouthwashes/therapeutic use , Administration, Topical , Adult , Aged , Bacterial Load/drug effects , Chlorhexidine/administration & dosage , Chronic Periodontitis/microbiology , Dental Plaque Index , Female , Follow-Up Studies , Gingivitis/drug therapy , Gram-Negative Aerobic Bacteria/drug effects , Gram-Negative Anaerobic Bacteria/drug effects , Gram-Positive Cocci/drug effects , Gram-Positive Rods/drug effects , Humans , Male , Middle Aged , Mouthwashes/administration & dosage , Periodontal Index , Periodontal Pocket/drug therapy , Periodontal Pocket/microbiology , Placebos , Porphyromonas/drug effects , Therapeutic Irrigation/methods
19.
J Biol Chem ; 285(50): 38811-7, 2010 Dec 10.
Article in English | MEDLINE | ID: mdl-20921229

ABSTRACT

The possible mechanism of casein aggregation and micelle buildup was studied in a new approach by letting α-casein adsorb from low concentration (0.1 mg·ml(-1)) solutions onto the charged surfaces of polyelectrolyte films. It was found that α-casein could adsorb onto both positively and negatively charged surfaces. However, only when its negative phosphoseryl clusters remained free, i.e. when it adsorbed onto a negative surface, could calcium phosphate (CaP) nanoclusters bind to the casein molecules. Once the CaP clusters were in place, step-by-step building of multilayered casein architectures became possible. The presence of CaP was essential; neither Ca(2+) nor phosphate could alone facilitate casein aggregation. Thus, it seems that CaP is the organizing motive in the casein micelle formation. Atomic force microscopy revealed that even a single adsorbed casein layer was composed of very small (in the range of tens of nanometers) spherical forms. The stiffness of the adsorbed casein layer largely increased in the presence of CaP. On this basis, we can imagine that casein micelles emerge according to the following scheme. The amphipathic casein monomers aggregate into oligomers via hydrophobic interactions even in the absence of CaP. Full scale, CaP-carrying micelles could materialize by interlocking these casein oligomers with CaP nanoclusters. Such a mechanism would not contradict former experimental results and could offer a synthesis between the submicelle and the block copolymer models of casein micelles.


Subject(s)
Caseins/chemistry , Adsorption , Animals , Biophysics/methods , Calcium/chemistry , Calcium Phosphates/chemistry , Cattle , Electrolytes/chemistry , Hydrophobic and Hydrophilic Interactions , Materials Testing , Micelles , Microscopy, Atomic Force/methods , Milk , Nanoparticles/chemistry , Protein Interaction Mapping , Spectroscopy, Fourier Transform Infrared , Surface Properties
20.
Biotechnol Bioeng ; 104(6): 1121-31, 2009 Dec 15.
Article in English | MEDLINE | ID: mdl-19634182

ABSTRACT

We report on elaboration of 12-mer peptides that reveal specific recognition for the following semiconductor (SC) surfaces: GaAs(100), InAs(100), GaN(0001), ZnSe(100), ZnTe(100), GaAs(111)A, GaSb(100), CdSe(100). A M13 bacteriophage library was used to screen 10(9) different 12-mer peptides against these substrates to finally isolate, in maximum six amplification cycles, peptides that bind to the target surfaces. The specific peptides for the InAs and ZnSe surfaces were obtained. Contrary, for the other SC surfaces several peptides with high affinities have been isolated. Aiming for a better specificity, when the phage display has been conducted through six cycles, the screening procedure got dominated by a phage present in the M13 bacteriophage library and the SVSVGMKPSPRP peptide has been selected for different SCs. The high amplification potential of this phage has been observed previously with different targets. Thus, precaution should be undertaken in defining adhesion peptides with the phage display technique and real affinity of the obtained biolinkers should be studied with other methods. We employed mass spectrometry (MALDI-TOF/TOF) to demonstrate the preferential attachment (or not) of the SVSVGMKPSPRP peptide to the different SC surfaces. This allows us to define a realistic selection of the expressed peptides presenting affinity for the studied eight SC surfaces. We demonstrate that with increasing the dielectric constants of the employed solvents, adhesion of the SVSVGMKPSPRP peptide onto GaN(0001) is hindered.


Subject(s)
Peptides/isolation & purification , Peptides/metabolism , Semiconductors , Bacteriophage M13/genetics , Peptide Library , Protein Binding , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , United States
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