Non-crystalline composite tissue engineering scaffolds using boron-containing bioactive glass and poly(d,l-lactic acid) coatings

International Nuclear Information System (INIS)

Mantsos, T; Chatzistavrou, X; Roether, J A; Boccaccini, A R; Hupa, L; Arstila, H

2009-01-01

The aim of this study was the fabrication of three-dimensional, highly porous, bioactive scaffolds using a recently developed bioactive glass powder, denominated '0106', with nominal composition (in wt%): 50 SiO 2 , 22.6 CaO, 5.9 Na 2 O, 4 P 2 O 5 , 12 K 2 O, 5.3 MgO and 0.2 B 2 O 3 . The optimum sintering conditions for the fabrication of scaffolds by the foam-replica method were identified (sintering temperature: 670 deg, C and dwell time: 5 h). Composite samples were also fabricated by applying a biopolymer coating of poly( D,L -lactic acid) (PDLLA) using a dip coating process. The average compressive strength values were 0.4 MPa for uncoated and 0.6 MPa for coated scaffolds. In vitro bioactivity studies in simulated body fluid (SBF) showed that a carbonate hydroxyapatite (HCAp) layer was deposited on uncoated and coated scaffolds after only 4 days of immersion in SBF, demonstrating the high in vitro bioactivity of the scaffolds. It was also confirmed that the scaffold structure remained amorphous (no crystallization) after the specific heat treatment used, with scaffolds exhibiting mechanical properties and bioactivity suitable for use in bone tissue engineering applications.

Non-crystalline composite tissue engineering scaffolds using boron-containing bioactive glass and poly(d,l-lactic acid) coatings

Energy Technology Data Exchange (ETDEWEB)

Mantsos, T; Chatzistavrou, X; Roether, J A; Boccaccini, A R [Department of Materials, Imperial College London, South Kensington Campus, London SW7 2AZ (United Kingdom); Hupa, L; Arstila, H, E-mail: a.boccaccini@imperial.ac.u [Process Chemistry Centre, Abo Akademi University, Piispankatu 8, FI-20500 Turku (Finland)

2009-10-15

The aim of this study was the fabrication of three-dimensional, highly porous, bioactive scaffolds using a recently developed bioactive glass powder, denominated '0106', with nominal composition (in wt%): 50 SiO{sub 2}, 22.6 CaO, 5.9 Na{sub 2}O, 4 P{sub 2}O{sub 5}, 12 K{sub 2}O, 5.3 MgO and 0.2 B{sub 2}O{sub 3}. The optimum sintering conditions for the fabrication of scaffolds by the foam-replica method were identified (sintering temperature: 670 deg, C and dwell time: 5 h). Composite samples were also fabricated by applying a biopolymer coating of poly({sub D,L}-lactic acid) (PDLLA) using a dip coating process. The average compressive strength values were 0.4 MPa for uncoated and 0.6 MPa for coated scaffolds. In vitro bioactivity studies in simulated body fluid (SBF) showed that a carbonate hydroxyapatite (HCAp) layer was deposited on uncoated and coated scaffolds after only 4 days of immersion in SBF, demonstrating the high in vitro bioactivity of the scaffolds. It was also confirmed that the scaffold structure remained amorphous (no crystallization) after the specific heat treatment used, with scaffolds exhibiting mechanical properties and bioactivity suitable for use in bone tissue engineering applications.

Structure, phases, and mechanical response of Ti-alloy bioactive glass composite coatings

Energy Technology Data Exchange (ETDEWEB)

Nelson, G.M.; Nychka, J.A. [Department of Chemical and Materials Engineering, University of Alberta, 7th Floor, Electrical and Computer Engineering Research Facility, Edmonton, Alberta T6G 2V4 (Canada); McDonald, A.G., E-mail: andre2@ualberta.ca [Department of Mechanical Engineering, University of Alberta, 4-9 Mechanical Engineering Building, Edmonton, Alberta T6G 2G8 (Canada)

2014-03-01

Porous titanium alloy-bioactive glass composite coatings were manufactured via the flame spray deposition process. The porous coatings, targeted for orthodontic and bone-fixation applications, were made from bioactive glass (45S5) powder blended with either commercially pure titanium (Cp-Ti) or Ti-6Al-4V alloy powder. Two sets of spray conditions, two metallic particle size distributions, and two glass particle size distributions were used for this study. Negative control coatings consisting of pure Ti-6Al-4V alloy or Cp-Ti were sprayed under both conditions. The as-sprayed coatings were characterized through quantitative optical cross-sectional metallography, X-ray diffraction (XRD), and ASTM Standard C633 tensile adhesion testing. Determination of the porosity and glassy phase distribution was achieved by using image analysis in accordance with ASTM Standard E2109. Theoretical thermodynamic and heat transfer modeling was conducted to explain experimental observations. Thermodynamic modeling was performed to estimate the flame temperature and chemical environment for each spray condition and a lumped capacitance heat transfer model was developed to estimate the temperatures attained by each particle. These models were used to establish trends among the choice of alloy, spray condition, and particle size distribution. The deposition parameters, alloy composition, and alteration of the feedstock powder size distribution had a significant effect on the coating microstructure, porosity, phases present, mechanical response, and theoretical particle temperatures that were attained. The most promising coatings were the Ti-6Al-4V-based composite coatings, which had bond strength of 20 ± 2 MPa (n = 5) and received reinforcement and strengthening from the inclusion of a glassy phase. It was shown that the use of the Ti-6Al-4V-bioactive glass composite coatings may be a superior choice due to the possible osteoproductivity from the bioactive glass, the potential ability to

Microstructures, hardness and bioactivity of hydroxyapatite coatings

CSIR Research Space (South Africa)

Tlotleng, Monnamme

2014-10-01

Full Text Available Hydroxyapatite (HAP) coatings on bioinert metals such as Ti–6Al–4V are necessary for biomedical applications. Together, HAP and Ti–6Al–4V are biocompatible and bioactive. The challenges of depositing HAP on Ti–6Al–4V with traditional thermal...

Bioactive hydroxyapatite/graphene composite coating and its corrosion stability in simulated body fluid

Energy Technology Data Exchange (ETDEWEB)

Janković, Ana; Eraković, Sanja [Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11 000 Belgrade (Serbia); Mitrić, Miodrag [Vinča Institute of Nuclear Sciences, University of Belgrade, Mike Petrovića Alasa 12-14, 11 000 Belgrade (Serbia); Matić, Ivana Z.; Juranić, Zorica D. [Institute of Oncology and Radiology of Serbia, Pasterova 14, 11 000 Belgrade (Serbia); Tsui, Gary C.P.; Tang, Chak-yin [Department of Industrial and Systems Engineering, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong (China); Mišković-Stanković, Vesna [Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11 000 Belgrade (Serbia); Rhee, Kyong Yop, E-mail: rheeky@khu.ac.kr [Department of Mechanical Engineering, Kyung Hee University, Yongin 449-701 (Korea, Republic of); Park, Soo Jin [Chemistry, College of Natural Sciences, Inha University, Incheon 402-751 (Korea, Republic of)

2015-03-05

Highlights: • Bioactive HAP/Gr coating on Ti was successfully obtained by EPD. • Increased fracture toughness of the HAP/Gr coating compared to pure HAP coating. • HAP/Gr coating exhibited superior biomimetic mineralization vs. pure HAP coating. • Gr improved the mechanical properties and thermal stability of HAP/Gr coating. • HAP/Gr coating was classified as non-cytotoxic against the targeted PBMC. - Abstract: The hydroxyapatite/graphene (HAP/Gr) composite was electrodeposited on Ti using the electrophoretic deposition process to obtain uniform bioactive coating with improved mechanical strength and favorable corrosion stability in simulated body fluid (SBF). Incorporation of Gr was verified by Raman spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, and X-ray photoelectron analysis. The HAP/Gr composite coating exhibited reduced surface cracks, nearly double the hardness, and elastic modulus increased by almost 50% compared to pure HAP coating, as estimated by a nanoindentation test. The bioactive HAP/Gr composite coating provided a newly formed apatite layer in SBF with enhanced corrosion stability, as evidenced by electrochemical impedance spectroscopy. The thermal stability of the HAP/Gr coating was improved in comparison to the pure HAP coating, and the Ca/P ratio was closer to the stoichiometric value. No antibacterial activity against Staphylococcus aureus or Escherichia coli could be verified. The HAP/Gr composite coating was classified as non-cytotoxic when tested against healthy peripheral blood mononuclear cells (PBMC)

Bioactive hydroxyapatite/graphene composite coating and its corrosion stability in simulated body fluid

International Nuclear Information System (INIS)

Janković, Ana; Eraković, Sanja; Mitrić, Miodrag; Matić, Ivana Z.; Juranić, Zorica D.; Tsui, Gary C.P.; Tang, Chak-yin; Mišković-Stanković, Vesna; Rhee, Kyong Yop; Park, Soo Jin

2015-01-01

Highlights: • Bioactive HAP/Gr coating on Ti was successfully obtained by EPD. • Increased fracture toughness of the HAP/Gr coating compared to pure HAP coating. • HAP/Gr coating exhibited superior biomimetic mineralization vs. pure HAP coating. • Gr improved the mechanical properties and thermal stability of HAP/Gr coating. • HAP/Gr coating was classified as non-cytotoxic against the targeted PBMC. - Abstract: The hydroxyapatite/graphene (HAP/Gr) composite was electrodeposited on Ti using the electrophoretic deposition process to obtain uniform bioactive coating with improved mechanical strength and favorable corrosion stability in simulated body fluid (SBF). Incorporation of Gr was verified by Raman spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, and X-ray photoelectron analysis. The HAP/Gr composite coating exhibited reduced surface cracks, nearly double the hardness, and elastic modulus increased by almost 50% compared to pure HAP coating, as estimated by a nanoindentation test. The bioactive HAP/Gr composite coating provided a newly formed apatite layer in SBF with enhanced corrosion stability, as evidenced by electrochemical impedance spectroscopy. The thermal stability of the HAP/Gr coating was improved in comparison to the pure HAP coating, and the Ca/P ratio was closer to the stoichiometric value. No antibacterial activity against Staphylococcus aureus or Escherichia coli could be verified. The HAP/Gr composite coating was classified as non-cytotoxic when tested against healthy peripheral blood mononuclear cells (PBMC)

Significant degradability enhancement in multilayer coating of polycaprolactone-bioactive glass/gelatin-bioactive glass on magnesium scaffold for tissue engineering applications

Energy Technology Data Exchange (ETDEWEB)

Yazdimamaghani, Mostafa [School of Chemical Engineering, Oklahoma State University, Stillwater, OK 74078 (United States); School of Materials Science and Engineering, Helmerich Advanced Technology Research Center, Oklahoma State University, Tulsa, OK 74106 (United States); Razavi, Mehdi [School of Materials Science and Engineering, Helmerich Advanced Technology Research Center, Oklahoma State University, Tulsa, OK 74106 (United States); Vashaee, Daryoosh [Electrical and Computer Engineering Department, North Carolina State University, Raleigh, NC 27606 (United States); Pothineni, Venkata Raveendra [Biomaterials and Advanced Drug Delivery Laboratory, Stanford University, Palo Alto, CA 94305 (United States); Rajadas, Jayakumar [Biomaterials and Advanced Drug Delivery Laboratory, Stanford University, Palo Alto, CA 94305 (United States); Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Palo Alto, CA 94305 (United States); Stanford Cardiovascular Institute, Stanford University School of Medicine, Palo Alto, CA 94305 (United States); Tayebi, Lobat, E-mail: lobat.tayebi@marquette.edu [School of Materials Science and Engineering, Helmerich Advanced Technology Research Center, Oklahoma State University, Tulsa, OK 74106 (United States); Biomaterials and Advanced Drug Delivery Laboratory, Stanford University, Palo Alto, CA 94305 (United States); Department of Developmental Sciences, Marquette University School of Dentistry, Milwaukee, WI 53233 (United States)

2015-05-30

Highlights: • PCL-BaG/Gel-BaG coating was applied on the surface of Mg scaffolds. • Mg scaffold/PCL-BaG/Gel-BaG presented improved biodegradation resistance. • Mg scaffold coated with the PCL-BaG layer indicated better bioactivity. - Abstract: Magnesium (Mg) is a promising candidate to be used in medical products especially as bone tissue engineering scaffolds. The main challenge for using Mg in biomedical applications is its high degradation rate in the body. For this reason, in this study, a multilayer polymeric layer composed of polycaprolactone (PCL) and gelatin (Gel) reinforced with bioactive glass (BaG) particles has been applied on the surface of Mg scaffolds. The materials characteristics of uncoated Mg scaffold, Mg scaffold coated only with PCL-BaG and Mg scaffold coated with PCL-BaG and Gel-BaG have been analyzed and compared in detail. Scanning electron microscope (SEM) equipped with energy dispersive spectroscopy (EDS), and Fourier transform infrared spectroscopy (FTIR) were utilized for microstructural studies. In vitro bioactivity and biodegradation evaluations were carried out by submerging the scaffolds in simulated body fluid (SBF) at pre-determined time points. The results demonstrated that Mg scaffold coated with PCL-BaG and Gel-BaG exhibited significant improvement in biodegradability.

Significant degradability enhancement in multilayer coating of polycaprolactone-bioactive glass/gelatin-bioactive glass on magnesium scaffold for tissue engineering applications

International Nuclear Information System (INIS)

Yazdimamaghani, Mostafa; Razavi, Mehdi; Vashaee, Daryoosh; Pothineni, Venkata Raveendra; Rajadas, Jayakumar; Tayebi, Lobat

2015-01-01

Highlights: • PCL-BaG/Gel-BaG coating was applied on the surface of Mg scaffolds. • Mg scaffold/PCL-BaG/Gel-BaG presented improved biodegradation resistance. • Mg scaffold coated with the PCL-BaG layer indicated better bioactivity. - Abstract: Magnesium (Mg) is a promising candidate to be used in medical products especially as bone tissue engineering scaffolds. The main challenge for using Mg in biomedical applications is its high degradation rate in the body. For this reason, in this study, a multilayer polymeric layer composed of polycaprolactone (PCL) and gelatin (Gel) reinforced with bioactive glass (BaG) particles has been applied on the surface of Mg scaffolds. The materials characteristics of uncoated Mg scaffold, Mg scaffold coated only with PCL-BaG and Mg scaffold coated with PCL-BaG and Gel-BaG have been analyzed and compared in detail. Scanning electron microscope (SEM) equipped with energy dispersive spectroscopy (EDS), and Fourier transform infrared spectroscopy (FTIR) were utilized for microstructural studies. In vitro bioactivity and biodegradation evaluations were carried out by submerging the scaffolds in simulated body fluid (SBF) at pre-determined time points. The results demonstrated that Mg scaffold coated with PCL-BaG and Gel-BaG exhibited significant improvement in biodegradability

Development of Bioactive Edible Coatings and Biodegradable Packaging Using Gamma Irradiation

International Nuclear Information System (INIS)

Lacroix, M.; Salmieri, S.

2010-01-01

Gamma irradiation was used to cross-link milk proteins in order to enhance the physico-chemical properties of edible films made of calcium caseinate, whey protein isolate and glycerol. Fourier Transform Infrared analysis was used to characterize the conformation of proteins adopted after irradiation. The molecular weight of cross-linked proteins was measured by Size-Exclusion Chromatography. Furthermore, the effect of the addition of methylcellulose to the irradiated protein matrix on the rheological properties (puncture strength, puncture deformation and water vapor permeability) of films was also studied. Moreover, cross-linking of polysaccharides under paste-like state was investigated and the cross-linking degree of the gel products was determined by gel fraction measurements and solubility percentage. In order to prepare bioactive coatings, several antifungal compounds were evaluated as bioactive compounds in order to select one of them to prepare an antimicrobial solution to spray onto strawberries or to encapsulate them in film formulations composed of milk proteins and methylcellulose based films. In addition, the bioactive coatings containing the antifungals were used to increase the radiosensitivity under air of moulds and total flora in strawberries and the relative sensitivity of selected formulations was calculated from their D10 value. The film formulation selected was used as a bioactive edible coating in order to determine their efficiency to increase the shelf life of fresh strawberries and to preserve their quality during storage. (author)

Electrophoretic deposition of ZnO/alginate and ZnO-bioactive glass/alginate composite coatings for antimicrobial applications

Energy Technology Data Exchange (ETDEWEB)

Cordero-Arias, L.; Cabanas-Polo, S.; Goudouri, O.M. [Institute of Biomaterials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Cauerstrasse 6, D-91058 Erlangen (Germany); Misra, S.K. [Materials Science and Engineering, Indian Institute of Technology Gandhinagar, Ahmedabad 382424 (India); Gilabert, J. [Institute of Ceramics Materials (ITC), University Jaume I, Avenida Vicent SosBaynat, 12006 Castellon (Spain); Valsami-Jones, E. [School of Geography, Earth and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT (United Kingdom); Sanchez, E. [Institute of Ceramics Materials (ITC), University Jaume I, Avenida Vicent SosBaynat, 12006 Castellon (Spain); Virtanen, S. [Institute for Surface Science and Corrosion (LKO, WW4), Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Martensstrasse 7, D-91058 Erlangen (Germany); Boccaccini, A.R., E-mail: aldo.boccaccini@ww.uni-erlangen.de [Institute of Biomaterials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Cauerstrasse 6, D-91058 Erlangen (Germany)

2015-10-01

Two organic/inorganic composite coatings based on alginate, as organic matrix, and zinc oxide nanoparticles (n-ZnO) with and without bioactive glass (BG), as inorganic components, intended for biomedical applications, were developed by electrophoretic deposition (EPD). Different n-ZnO (1–10 g/L) and BG (1–1.5 g/L) contents were studied for a fixed alginate concentration (2 g/L). The presence of n-ZnO was confirmed to impart antibacterial properties to the coatings against gram-negative bacteria Escherichia coli, while the BG induced the formation of hydroxyapatite on coating surfaces thereby imparting bioactivity, making the coating suitable for bone replacement applications. Coating composition was analyzed by thermogravimetric analysis (TG), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and energy dispersive X-ray spectroscopy (EDS) analyses. Scanning electron microscopy (SEM) was employed to study both the surface and the cross section morphology of the coatings. Polarization curves of the coated substrates made in cell culture media at 37 °C confirmed the corrosion protection function of the novel organic/inorganic composite coatings. - Highlights: • Organic–inorganic nanocomposite coatings fabricated by electrophoretic deposition • nZnO and bioactive glass containing alginate coatings exhibit antibacterial effect. • Bioactive character and anticorrosion function of coatings demonstrated.

Bioactive glass coatings for orthopedic metallic implants

Energy Technology Data Exchange (ETDEWEB)

Lopez-Esteban, Sonia; Saiz, Eduardo; Fujino, Sigheru; Oku, Takeo; Suganuma, Katsuaki; Tomsia, Antoni P.

2003-06-30

The objective of this work is to develop bioactive glass coatings for metallic orthopedic implants. A new family of glasses in the SiO2-Na2O-K2O-CaO-MgO-P2O5 system has been synthesized and characterized. The glass properties (thermal expansion, softening and transformation temperatures, density and hardness) are in line with the predictions of established empirical models. The optimized firing conditions to fabricate coatings on Ti-based and Co-Cr alloys have been determined and related to the glass properties and the interfacial reactions. Excellent adhesion to alloys has been achieved through the formation of 100-200 nm thick interfacial layers (Ti5Si3 on Ti-based alloys and CrOx on Co-Cr). Finally, glass coatings, approximately 100 mu m thick, have been fabricated onto commercial Ti alloy-based dental implants.

Electrophoretic deposition of chitosan/45S5 bioactive glass composite coatings doped with Zn and Sr

Directory of Open Access Journals (Sweden)

Marta eMiola

2015-10-01

Full Text Available In this research work the original 45S5 bioactive glass (BG was modified by introducing zinc and/or strontium oxide (6% mol in place of calcium oxide. Sr was added for its ability to stimulate bone formation, Zn for its role in bone metabolism, antibacterial properties and anti-inflammatory effect. The glasses were produced by means of melting and quenching process. SEM and XRD analyses evidenced that Zr and Sr introduction did not modify the glass structure and morphology, while compositional analysis (EDS demonstrated the effective addition of these elements inside the glass network. Bioactivity test in simulated body fluid (SBF up to one month evidenced a reduced bioactivity kinetics for Zn-doped glasses. Doped glasses were combined with chitosan to produce organic/inorganic composite coatings on stainless steel AISI 316L by electrophoretic deposition (EPD. Two EPD processes were considered for coating development, namely direct current EPD (DC-EPD and alternating current EPD (AC-EPD. The stability of the suspension was analysed and the deposition parameters were optimized. Tape and bending tests demonstrated a good coating-substrate adhesion for coatings containing 45S5-Sr and 45S5-ZnSr glasses, while the adhesion to the substrate decreased by using 45S5-Zn glass. FTIR analyses demonstrated the composite nature of coatings and SEM observations indicated that glass particles were well integrated in the polymeric matrix, the coatings were fairly homogeneous and free of cracks; moreover the AC-EPD technique provided better results than DC-EPD in terms of coating quality. SEM, XRD analyses and Raman spectroscopy, performed after bioactivity test in SBF solution, confirmed the bioactive behaviour of 45S5-Sr containing coating, while coatings containing Zn exhibited no hydroxyapatite formation.

Bioactive Coatings for Orthopaedic Implants—Recent Trends in Development of Implant Coatings

Directory of Open Access Journals (Sweden)

Bill G. X. Zhang

2014-07-01

Full Text Available Joint replacement is a major orthopaedic procedure used to treat joint osteoarthritis. Aseptic loosening and infection are the two most significant causes of prosthetic implant failure. The ideal implant should be able to promote osteointegration, deter bacterial adhesion and minimize prosthetic infection. Recent developments in material science and cell biology have seen the development of new orthopaedic implant coatings to address these issues. Coatings consisting of bioceramics, extracellular matrix proteins, biological peptides or growth factors impart bioactivity and biocompatibility to the metallic surface of conventional orthopaedic prosthesis that promote bone ingrowth and differentiation of stem cells into osteoblasts leading to enhanced osteointegration of the implant. Furthermore, coatings such as silver, nitric oxide, antibiotics, antiseptics and antimicrobial peptides with anti-microbial properties have also been developed, which show promise in reducing bacterial adhesion and prosthetic infections. This review summarizes some of the recent developments in coatings for orthopaedic implants.

Bioactivity and biocompatibility of hydroxyapatite-based bioceramic coatings on zirconium by plasma electrolytic oxidation

Energy Technology Data Exchange (ETDEWEB)

Aktuğ, Salim Levent, E-mail: saktug@gtu.edu.tr [The Department of Materials Science and Engineering, Gebze Technical University, Gebze, Kocaeli 41400 (Turkey); Durdu, Salih, E-mail: durdusalih@gmail.com [The Department of Industrial Engineering, Giresun University, Merkez, Giresun 28200 (Turkey); Yalçın, Emine, E-mail: emine.yalcin@giresun.edu.tr [The Department of Biology, Giresun University, Merkez, Giresun 28200 (Turkey); Çavuşoğlu, Kültigin, E-mail: kultigin.cavusoglu@giresun.edu.tr [The Department of Biology, Giresun University, Merkez, Giresun 28200 (Turkey); Usta, Metin, E-mail: ustam@gtu.edu.tr [The Department of Materials Science and Engineering, Gebze Technical University, Gebze, Kocaeli 41400 (Turkey); Materials Institute, Marmara Research Center, TUBITAK, Gebze, Kocaeli 41470 (Turkey)

2017-02-01

In the present work, hydroxyapatite (HAP)-based plasma electrolytic oxide (PEO) coatings were produced on zirconium at different current densities in a solution containing calcium acetate and β-calcium glycerophosphate by a single step. The phase structure, surface morphology, functional groups, thickness and roughness of the coatings were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), eddy current method and surface profilometer, respectively. The phases of cubic-zirconia, calcium zirconate and HAP were detected by XRD. The amount of HAP and calcium zirconate increased with increasing current density. The surface of the coatings was very porous and rough. Moreover, bioactivity and biocompatibility of the coatings were analyzed in vitro immersion simulated body fluid (SBF) and MTT (3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyl tetrazolium bromide) assay, hemolysis assay and bacterial formation. The apatite-forming ability of the coatings was evaluated after immersion in SBF up to 28 days. After immersion, the bioactivity of HAP-based coatings on zirconium was greater than the ones of uncoated zirconium and zirconium oxide-based surface. The bioactivity of PEO surface on zirconium was significantly improved under SBF conditions. The bacterial adhesion of the coatings decreased with increasing current density. The bacterial adhesion of the coating produced at 0.370 A/cm{sup 2} was minimum compared to uncoated zirconium coated at 0.260 and 0.292 A/cm{sup 2}. The hemocompatibility of HAP-based surfaces was improved by PEO. The cell attachment and proliferation of the PEO coatings were better than the one of uncoated zirconium according to MTT assay results. - Highlights: • Hydroxyapatite was formed on zirconium at different current densities by single-step plasma electrolytic oxidation. • The amount of hydroxyapatite and calcium-based phases increased with

Bioactivity and biocompatibility of hydroxyapatite-based bioceramic coatings on zirconium by plasma electrolytic oxidation.

Science.gov (United States)

Aktuğ, Salim Levent; Durdu, Salih; Yalçın, Emine; Çavuşoğlu, Kültigin; Usta, Metin

2017-02-01

In the present work, hydroxyapatite (HAP)-based plasma electrolytic oxide (PEO) coatings were produced on zirconium at different current densities in a solution containing calcium acetate and β-calcium glycerophosphate by a single step. The phase structure, surface morphology, functional groups, thickness and roughness of the coatings were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), eddy current method and surface profilometer, respectively. The phases of cubic-zirconia, calcium zirconate and HAP were detected by XRD. The amount of HAP and calcium zirconate increased with increasing current density. The surface of the coatings was very porous and rough. Moreover, bioactivity and biocompatibility of the coatings were analyzed in vitro immersion simulated body fluid (SBF) and MTT (3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyl tetrazolium bromide) assay, hemolysis assay and bacterial formation. The apatite-forming ability of the coatings was evaluated after immersion in SBF up to 28days. After immersion, the bioactivity of HAP-based coatings on zirconium was greater than the ones of uncoated zirconium and zirconium oxide-based surface. The bioactivity of PEO surface on zirconium was significantly improved under SBF conditions. The bacterial adhesion of the coatings decreased with increasing current density. The bacterial adhesion of the coating produced at 0.370A/cm 2 was minimum compared to uncoated zirconium coated at 0.260 and 0.292A/cm 2 . The hemocompatibility of HAP-based surfaces was improved by PEO. The cell attachment and proliferation of the PEO coatings were better than the one of uncoated zirconium according to MTT assay results. Copyright © 2016 Elsevier B.V. All rights reserved.

Enhanced bioactivity and biocompatibility of nanostructured hydroxyapatite coating by hydrothermal annealing

International Nuclear Information System (INIS)

Hahn, Byung-Dong; Lee, Jeong-Min; Park, Dong-Soo; Choi, Jong-Jin; Ryu, Jungho; Yoon, Woon-Ha; Choi, Joon-Hwan; Lee, Byoung-Kuk; Kim, Jong-Woo; Kim, Hyoun-Ee; Kim, Seong-Gon

2011-01-01

The crystallinity of hydroxyapatite (HA) coatings prepared by aerosol deposition may be increased by heating in air or low-temperature hydrothermal processing. From the X-ray diffraction and Fourier transform infrared spectroscopy results, it was revealed that the crystallinity of the HA coatings significantly increased after the post-annealing. Transmission electron microscopy showed that the conventional furnace heating induced the substantial growth of the HA crystallites, whereas the hydrothermal treatment did not bring about any remarkable change in the HA crystallite size, which remained below 20 nm. The bioactivity of the HA coatings was estimated by the acellular simulated body fluid immersion test. After immersion for 7 days, newly-precipitated apatite crystals were only observed on the surfaces of the samples hydrothermally treated at 170 and 190 deg. C. In addition, the alkaline phosphatase activity of MC3T3-E1 preosteoblast cells cultured on the hydrothermally treated samples was significantly higher than those on the as-deposited coating and conventional furnace heated samples. The enhanced bioactivity and excellent biological in vitro cellular response of the hydrothermally treated samples were attributed to their nanostructured nature and high degree of crystallinity.

Neocellularization and neovascularization of nanosized bioactive glass-coated decellularized trabecular bone scaffolds

KAUST Repository

Gerhardt, Lutz Christian

2012-09-11

In this study, the in vivo recellularization and neovascularization of nanosized bioactive glass (n-BG)-coated decellu-larized trabecular bone scaffolds were studied in a rat model and quantified using stereological analyses. Based on the highest amount of vascular endothelial growth factor (VEGF) secreted by human fibroblasts grown on n-BG coatings (0-1.245 mg/cm 2), decellularized trabecular bone samples (porosity: 43-81%) were coated with n-BG particles. Grown on n-BG particles at a coating density of 0.263 mg/cm2, human fibroblasts produced 4.3 times more VEGF than on uncoated controls. After 8 weeks of implantation in Sprague-Dawley rats, both uncoated and n-BG-coated samples were well infiltrated with newly formed tissue (47-48%) and blood vessels (3-4%). No significant differences were found in cellularization and vascularization between uncoated bone scaffolds and n-BG-coated scaffolds. This finding indicates that the decellularized bone itself may exhibit growth-promoting properties induced by the highly interconnected pore microarchitecture and/or proteins left behind on decellularized scaffolds. Even if we did not find proangiogenic effects in n-BG-coated bone scaffolds, a bioactive coating is considered to be beneficial to impart osteoinductive and osteoconductive properties to decellularized bone. n-BG-coated bone grafts have thus high clinical potential for the regeneration of complex tissue defects given their ability for recellularization and neovascularization. © 2012 Wiley Periodicals, Inc.

Dual antagonists of integrins.

Science.gov (United States)

Nadrah, K; Dolenc, M Sollner

2005-01-01

The roles of integrins in pathologies have been studied intensively and only partially explained. This has resulted in the development of several nanomolar antagonists to certain integrins. In most cases, the aim was to produce compounds which are highly selective towards specific integrins. This paradigm has recently shifted a little. Targeting two or more integrins with one compound has become a very attractive concept, especially since it has become clear that several severe disorders, such as pathological angiogenesis, cannot be treated just with highly specific integrin antagonists. This review is aimed to elucidate some aspects regarding the design of drugs with dual activity towards integrins. Integrin structure and tissue distribution will first be described, in order to provide the basis for their functions in various pathologies which will follow. Inhibitors of several pairs of integrins will be described.

Nanocrystalline β-Ta Coating Enhances the Longevity and Bioactivity of Medical Titanium Alloys

Directory of Open Access Journals (Sweden)

Linlin Liu

2016-09-01

Full Text Available A β-Ta nanocrystalline coating was engineered onto a Ti-6Al-4V substrate using a double cathode glow discharge technique to improve the corrosion resistance and bioactivity of this biomedical alloy. The new coating has a thickness of ~40 μm and exhibits a compact and homogeneous structure composed of equiaxed β-Ta grains with an average grain size of ~22 nm, which is well adhered on the substrate. Nanoindentation and scratch tests indicated that the β-Ta coating exhibited high hardness combined with good resistance to contact damage. The electrochemical behavior of the new coating was systematically investigated in Hank’s physiological solution at 37 °C. The results showed that the β-Ta coating exhibited a superior corrosion resistance as compared to uncoated Ti-6Al-4V and commercially pure tantalum, which was attributed to a stable passive film formed on the β-Ta coating. The in vitro bioactivity was studied by evaluating the apatite-forming capability of the coating after seven days of immersion in Hank’s physiological solution. The β-Ta coating showed a higher apatite-forming ability than both uncoated Ti-6Al-4V and commercially pure Ta, suggesting that the β-Ta coating has the potential to enhance functionality and increase longevity of orthopaedic implants.

Structure and in vitro bioactivity of ceramic coatings on magnesium alloys by microarc oxidation

Science.gov (United States)

Yu, Huijun; Dong, Qing; Dou, Jinhe; Pan, Yaokun; Chen, Chuanzhong

2016-12-01

Magnesium and its alloys have the potential to serve as lightweight, degradable, biocompatible and bioactive orthopedic implants for load-bearing applications. However, severe local corrosion attack and high corrosion rate have prevented their further clinical use. Micro-arc oxidation (MAO) is proved to be a simple, controllable and efficient electrochemistry technique that can prepare protective ceramic coatings on magnesium alloys. In this paper, electrolyte containing silicate salts was used for microarc oxidation to form ceramic bioactive coatings on the ZK61 alloy substrate. The structure characteristics and element distributions of the coating were investigated by XRD, TEM, SEM and EPMA. The MAO samples were immersed in simulated body fluid (SBF) for 7 and 14 days, respectively. The surface characteristic of the immersed coatings was investigated by Fourier-transform infrared (FTIR) spectroscopy. The results show that these MAO coatings have low crystallinity and are mainly composed of MgO, Mg2SiO4 and Mg2Si2O6. The coating surface is porous. During the SBF immersion period, the nucleation and precipitation of bone-like apatites occur on the MAO coating surface. The corrosion resistance of the substrate is improved by the MAO coatings.

Adhesive Bioactive Coatings Inspired by Sea Life.

Science.gov (United States)

Rego, Sónia J; Vale, Ana C; Luz, Gisela M; Mano, João F; Alves, Natália M

2016-01-19

Inspired by nature, in particular by the marine mussels adhesive proteins (MAPs) and by the tough brick-and-mortar nacre-like structure, novel multilayered films are prepared in the present work. Organic-inorganic multilayered films, with an architecture similar to nacre based on bioactive glass nanoparticles (BG), chitosan, and hyaluronic acid modified with catechol groups, which are the main components responsible for the outstanding adhesion in MAPs, are developed for the first time. The biomimetic conjugate is prepared by carbodiimide chemistry and analyzed by ultraviolet-visible spectrophotometry. The buildup of the multilayered films is monitored with a quartz crystal microbalance with dissipation monitoring, and their topography is characterized by atomic force microscopy. The mechanical properties reveal that the films containing catechol groups and BG present an enhanced adhesion. Moreover, the bioactivity of the films upon immersion in a simulated body fluid solution is evaluated by scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and X-ray diffraction. It was found that the constructed films promote the formation of bonelike apatite in vitro. Such multifunctional mussel inspired LbL films, which combine enhanced adhesion and bioactivity, could be potentially used as coatings of a variety of implants for orthopedic applications.

Significant degradability enhancement in multilayer coating of polycaprolactone-bioactive glass/gelatin-bioactive glass on magnesium scaffold for tissue engineering applications

Science.gov (United States)

Yazdimamaghani, Mostafa; Razavi, Mehdi; Vashaee, Daryoosh; Pothineni, Venkata Raveendra; Rajadas, Jayakumar; Tayebi, Lobat

2015-05-01

Magnesium (Mg) is a promising candidate to be used in medical products especially as bone tissue engineering scaffolds. The main challenge for using Mg in biomedical applications is its high degradation rate in the body. For this reason, in this study, a multilayer polymeric layer composed of polycaprolactone (PCL) and gelatin (Gel) reinforced with bioactive glass (BaG) particles has been applied on the surface of Mg scaffolds. The materials characteristics of uncoated Mg scaffold, Mg scaffold coated only with PCL-BaG and Mg scaffold coated with PCL-BaG and Gel-BaG have been analyzed and compared in detail. Scanning electron microscope (SEM) equipped with energy dispersive spectroscopy (EDS), and Fourier transform infrared spectroscopy (FTIR) were utilized for microstructural studies. In vitro bioactivity and biodegradation evaluations were carried out by submerging the scaffolds in simulated body fluid (SBF) at pre-determined time points. The results demonstrated that Mg scaffold coated with PCL-BaG and Gel-BaG exhibited significant improvement in biodegradability.

Combined PIXE and SEM study of the behaviour of trace elements in gel formed around implant coated with bioactive glass

Science.gov (United States)

Oudadesse, H.; Irigaray, J. L.; Barbotteau, Y.; Brun, V.; Moretto, Ph.

2002-05-01

Bioactive glasses are used as coating biomaterials to facilitate anchorage of metallic prostheses implanted into the body. The aim of this work is to study the behavior of gel formed in contact with alloys and BVA and BVH bioactive glasses implanted. Cylinders of metallic implants composed by Ti, Al and V, are coated with bioactive glass. Three sheep were implanted for different time length: 3, 6 and 12 months in the femoral epiphysis. Results obtained with particle induced X-ray emission and scanning electron microscopy show that BVA coating induces a better contact between the metallic implant and bone. On the other hand, BVH coating prevents corrosion from the metallic implant.

Effect of edible coatings on bioactive compounds and antioxidant capacity of tomatoes at different maturity stages.

Science.gov (United States)

Dávila-Aviña, Jorge E; Villa-Rodríguez, José A; Villegas-Ochoa, Mónica A; Tortoledo-Ortiz, Orlando; Olivas, Guadalupe I; Ayala-Zavala, J Fernando; González-Aguilar, Gustavo A

2014-10-01

This work evaluated the effect of carnauba and mineral oil coatings on the bioactive compounds and antioxidant capacity of tomato fruits (cv. "Grandela"). Carnauba and mineral oil coatings were applied on fresh tomatoes at two maturity stages (breaker and pink) over 28 day of storage at 10 °C was evaluated. Bioactive compound and antioxidant activity assays included total phenols, total flavonoids, ascorbic acid (ASA), lycopene, DPPH radical scavenging activity (%RSA), trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity assay (ORAC). The total phenolic, flavonoid and lycopene contents were significantly lower for coated fruit than control fruits. However, ascorbic acid content was highest in fruits treated with carnauba, followed by mineral oil coating and control fruits. The ORAC values were highest in breaker tomatoes coated with carnauba wax, followed by mineral oil-coated fruits and controls. No significant differences in ORAC values were observed in pink tomatoes. % RSA and TEAC values were higher for controls than for coated fruit. Edible coatings preserve the overall quality of tomatoes during storage without affecting the nutritional quality of fruit. We found that the physiological response to the coatings is in function of the maturity stage of tomatoes. The information obtained in this study support to use of edible coating as a safe and good alternative to preserve tomato quality, and that the changes of bioactive compounds and antioxidant activity of tomato fruits, was not negatively affected. This approach can be used by producers to preserve tomato quality.

Improved bonding strength of bioactive cermet Cold Gas Spray coatings.

Science.gov (United States)

Gardon, M; Concustell, A; Dosta, S; Cinca, N; Cano, I G; Guilemany, J M

2014-12-01

The fabrication of cermet biocompatible coatings by means Cold Gas Spray (CGS) provides prosthesis with outstanding mechanical properties and the required composition for enhancing the bioactivity of prosthetic materials. In this study, hydroxyapatite/Titanium coatings were deposited by means of CGS technology onto titanium alloy substrates with the aim of building-up well-bonded homogeneous coatings. Powders were blended in different percentages and sprayed; as long as the amount of hydroxyapatite in the feedstock increased, the quality of the coating was reduced. Besides, the relation between the particle size distribution of ceramic and metallic particles is of significant consideration. Plastic deformation of titanium particles at the impact eased the anchoring of hard hydroxyapatite particles present at the top surface of the coating, which assures the looked-for interaction with the cells. Coatings were immersed in Hank's solution for 1, 4 and 7 days; bonding strength value was above 60 MPa even after 7 days, which enhances common results of HAp coatings obtained by conventional thermal spray technologies. Copyright © 2014 Elsevier B.V. All rights reserved.

Effect of different coatings on post-harvest quality and bioactive compounds of pomegranate (Punica granatum L.) fruits.

Science.gov (United States)

Meighani, Hossein; Ghasemnezhad, Mahmood; Bakhshi, Davood

2015-07-01

The effect of three different coatings; resin wax (Britex Ti), carnauba wax (Xedasol M14), and chitosan (1 and 2 % w/v) on postharvest quality of pomegranate fruits were investigated. Fruits quality characteristics and bioactive compounds were evaluated during 40, 80 and 120 days storage at 4.5 °C and 3 additional days at 20 °C. The results showed that uncoated fruits showed higher respiration rate, weight loss, L* and b* values of arils, total soluble solids (TSS)/titratable acidity (TA), and pH than coated fruits during storage. Coating treatments could delay declining TSS and TA percent, a* value of arils, as well as bioactive compounds such as total phenolics, flavonoids and anthocyanins content and antioxidant activity. The coated fruits with commercial resin and carnauba waxes showed significantly lower respiration rate and weight loss than other treatments, however carnauba wax could maintain considerably higher fruits quality and bioactive compounds than other coating treatments. The results suggested that postharvest application of carnauba wax have a potential to extend storage life of pomegranate fruits by reducing respiration rate, water loss and maintaining fruit quality.

Microstructures, hardness and bioactivity of hydroxyapatite coatings deposited by direct laser melting process

International Nuclear Information System (INIS)

Tlotleng, Monnamme; Akinlabi, Esther; Shukla, Mukul; Pityana, Sisa

2014-01-01

Hydroxyapatite (HAP) coatings on bioinert metals such as Ti–6Al–4V are necessary for biomedical applications. Together, HAP and Ti–6Al–4V are biocompatible and bioactive. The challenges of depositing HAP on Ti–6Al–4V with traditional thermal spraying techniques are well founded. In this paper, HAP was coated on Ti–6Al–4V using direct laser melting (DLM) process. This process, unlike the traditional coating processes, is able to achieve coatings with good metallurgical bonding and little dilution. The microstructural and mechanical properties, chemical composition and bio-activities of the produced coatings were studied with optical microscopy, scanning electron microscope equipped with energy dispersive X-ray spectroscopy, and Vickers hardness machine, and by immersion test in Hanks' solution. The results showed that the choice of the laser power has much influence on the evolving microstructure, the mechanical properties and the retainment of HAP on the surface of the coating. Also, the choice of laser power of 750 W led to no dilution. The microhardness results inferred a strong intermetallic–ceramic interfacial bonding; which meant that the 750 W coating could survive long in service. Also, the coating was softer at the surface and stronger in the heat affected zones. Hence, this process parameter setting can be considered as an optimal setting. The soak tests revealed that the surface of the coating had unmelted crystals of HAP. The CaP ratio conducted on the soaked coating was 2.00 which corresponded to tetra calcium phosphate. This coating seems attractive for metallic implant applications. - Highlights: • Characteristics of HAP coatings produced on Ti-6Al-4V achieved with direct laser melting are reported. • Optimal process parameters necessary to achieve biocompatible coating are reported. • The SEM micrograph of the soaked HAP coating revealed partially melted crystals of HAP. • The HAP coating was retained at the surface of

Microstructures, hardness and bioactivity of hydroxyapatite coatings deposited by direct laser melting process

Energy Technology Data Exchange (ETDEWEB)

Tlotleng, Monnamme, E-mail: MTlotleng@csir.co.za [Laser Materials Processing Group, National Laser Center CSIR, Pretoria 0001 (South Africa); Department of Mechanical Engineering Science, University of Johannesburg, Auckland Park, Kingsway Campus, Johannesburg 2006 (South Africa); Akinlabi, Esther [Department of Mechanical Engineering Science, University of Johannesburg, Auckland Park, Kingsway Campus, Johannesburg 2006 (South Africa); Shukla, Mukul [Department of Mechanical Engineering Technology, University of Johannesburg, Doornfontein Campus, Johannesburg 2006 (South Africa); Department of Mechanical Engineering, MNNIT, Allahabad, UP 211004 (India); Pityana, Sisa [Laser Materials Processing Group, National Laser Center CSIR, Pretoria 0001 (South Africa); Department of Chemical and Metallurgical Engineering, Tshwane University of Technology, Pretoria 0001 (South Africa)

2014-10-01

Hydroxyapatite (HAP) coatings on bioinert metals such as Ti–6Al–4V are necessary for biomedical applications. Together, HAP and Ti–6Al–4V are biocompatible and bioactive. The challenges of depositing HAP on Ti–6Al–4V with traditional thermal spraying techniques are well founded. In this paper, HAP was coated on Ti–6Al–4V using direct laser melting (DLM) process. This process, unlike the traditional coating processes, is able to achieve coatings with good metallurgical bonding and little dilution. The microstructural and mechanical properties, chemical composition and bio-activities of the produced coatings were studied with optical microscopy, scanning electron microscope equipped with energy dispersive X-ray spectroscopy, and Vickers hardness machine, and by immersion test in Hanks' solution. The results showed that the choice of the laser power has much influence on the evolving microstructure, the mechanical properties and the retainment of HAP on the surface of the coating. Also, the choice of laser power of 750 W led to no dilution. The microhardness results inferred a strong intermetallic–ceramic interfacial bonding; which meant that the 750 W coating could survive long in service. Also, the coating was softer at the surface and stronger in the heat affected zones. Hence, this process parameter setting can be considered as an optimal setting. The soak tests revealed that the surface of the coating had unmelted crystals of HAP. The CaP ratio conducted on the soaked coating was 2.00 which corresponded to tetra calcium phosphate. This coating seems attractive for metallic implant applications. - Highlights: • Characteristics of HAP coatings produced on Ti-6Al-4V achieved with direct laser melting are reported. • Optimal process parameters necessary to achieve biocompatible coating are reported. • The SEM micrograph of the soaked HAP coating revealed partially melted crystals of HAP. • The HAP coating was retained at the surface of

Osseointegration properties of titanium dental implants modified with a nanostructured coating based on ordered porous silica and bioactive glass nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Covarrubias, Cristian, E-mail: ccovarrubias@odontologia.uchile.cl [Laboratory of Nanobiomaterials, Institute for Research in Dental Sciences, Faculty of Dentistry, University of Chile, Santiago (Chile); Mattmann, Matías [Laboratory of Nanobiomaterials, Institute for Research in Dental Sciences, Faculty of Dentistry, University of Chile, Santiago (Chile); Von Marttens, Alfredo [Department of Prosthesis, Faculty of Dentistry, University of Chile, Santiago (Chile); Caviedes, Pablo; Arriagada, Cristián [Laboratory of Cell Therapy, ICBM, Faculty of Medicine, University of Chile (Chile); Valenzuela, Francisco [Laboratory of Nanobiomaterials, Institute for Research in Dental Sciences, Faculty of Dentistry, University of Chile, Santiago (Chile); Rodríguez, Juan Pablo [Laboratory of Cell Biology, INTA, University of Chile, Santiago (Chile); Corral, Camila [Department of Restorative Dentistry, Faculty of Dentistry, University of Chile, Santiago (Chile)

2016-02-15

Graphical abstract: - Highlights: • The fabrication of a coating for osseointegration of titanium implant is presented. • The coating consists of nanoporous silica loaded with bioactive glass nanoparticles. • Coating accelerates the in vitro formation of apatite in simulated body fluid. • Coating promotes the osteogenic differentiation of stem cells. • Coating accelerates the formation of bone tissue in the periphery of the implant. - Abstract: The fabrication of a nanoporous silica coating loaded with bioactive glass nanoparticles (nBG/NSC) on titanium dental implant surface and its in vitro and in vivo evaluation is presented. The coating was produced by a combined sol–gel and evaporation induced self-assembly process. In vitro bioactivity was assessed in simulated body fluid (SBF) and investigating the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). A rat tibial model was employed to analyze the bone response to nBG/NSC-modified titanium implant surface in vivo. The nBG/NSC coating was confirmed at nano level to be constituted by a highly ordered nanoporous silica structure. The coating nanotopography in conjunction with the bioactivity of the BG particles accelerate the in vitro apatite formation and promote the osteogenic differentiation of hBMSCs in absence of osteogenic supplements. These properties accelerate the formation of bone tissue in the periphery of the implant after 3 weeks of implantation. Backscattered scanning electron microscopy images revealed the presence of gaps and soft tissue in the unmodified implant after 6 weeks, whereas the nBG/NSC-modified implant showed mature bone in intimate contact with the implant surface. The nBG/NSC coating appears promising for accelerating the osseointegration of dental implants.

Osseointegration properties of titanium dental implants modified with a nanostructured coating based on ordered porous silica and bioactive glass nanoparticles

International Nuclear Information System (INIS)

Covarrubias, Cristian; Mattmann, Matías; Von Marttens, Alfredo; Caviedes, Pablo; Arriagada, Cristián; Valenzuela, Francisco; Rodríguez, Juan Pablo; Corral, Camila

2016-01-01

Graphical abstract: - Highlights: • The fabrication of a coating for osseointegration of titanium implant is presented. • The coating consists of nanoporous silica loaded with bioactive glass nanoparticles. • Coating accelerates the in vitro formation of apatite in simulated body fluid. • Coating promotes the osteogenic differentiation of stem cells. • Coating accelerates the formation of bone tissue in the periphery of the implant. - Abstract: The fabrication of a nanoporous silica coating loaded with bioactive glass nanoparticles (nBG/NSC) on titanium dental implant surface and its in vitro and in vivo evaluation is presented. The coating was produced by a combined sol–gel and evaporation induced self-assembly process. In vitro bioactivity was assessed in simulated body fluid (SBF) and investigating the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). A rat tibial model was employed to analyze the bone response to nBG/NSC-modified titanium implant surface in vivo. The nBG/NSC coating was confirmed at nano level to be constituted by a highly ordered nanoporous silica structure. The coating nanotopography in conjunction with the bioactivity of the BG particles accelerate the in vitro apatite formation and promote the osteogenic differentiation of hBMSCs in absence of osteogenic supplements. These properties accelerate the formation of bone tissue in the periphery of the implant after 3 weeks of implantation. Backscattered scanning electron microscopy images revealed the presence of gaps and soft tissue in the unmodified implant after 6 weeks, whereas the nBG/NSC-modified implant showed mature bone in intimate contact with the implant surface. The nBG/NSC coating appears promising for accelerating the osseointegration of dental implants.

Cellulose acetate/hydroxyapatite/chitosan coatings for improved corrosion resistance and bioactivity

Energy Technology Data Exchange (ETDEWEB)

Zhong, Zhenyu; Qin, Jinli [Advanced Biomaterials and Tissue Engineering Center, Huazhong University of Science and Technology, Wuhan 430074 (China); Ma, Jun, E-mail: caltary@gmail.com [Advanced Biomaterials and Tissue Engineering Center, Huazhong University of Science and Technology, Wuhan 430074 (China); Department of Biomedical Engineering, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074 (China)

2015-04-01

Cellulose acetate (CA) nanofibers were deposited on stainless steel plates by electrospinning technique. The composite of hydroxyapatite (HAP) nanoparticles and chitosan (CHI) was coated subsequently by dip-coating. The structure and morphology of the obtained coatings were investigated by Fourier transform infrared spectroscopy and scanning electron microscopy. The stability of the coatings in physiological environment was studied using electrochemical polarization and impedance spectroscopy. The CA nanofibers were embedded in the HAP/CHI coating and the resulted composite film was densely packed and uniform on the substrate. The in vitro biomineralization study of the coated samples immersed in simulated body fluid (SBF) confirmed the formation ability of bone-like apatite layer on the surface of HAP-containing coatings. Furthermore, the coatings could provide corrosion resistance to the stainless steel substrate in SBF. The electrochemical results suggested that the incorporation of CA nanofibers could improve the corrosion resistance of the HAP/CHI coating. Thus, biocompatible CA/HAP/CHI coated metallic implants could be very useful in the long-term stability of the biomedical applications. - Highlights: • The composite coatings were prepared by electrospinning and dip-coating. • Good in vitro bioactivity of the CA/HAP/CHI coating was confirmed. • Electrochemical behaviors in SBF of the coatings have been studied. • The CA/HAP/CHI coating shows better resistance property than HAP/CHI.

Utility of tantalum (Ta) coating to improve surface hardness in vitro bioactivity and biocompatibility of Co–Cr

Energy Technology Data Exchange (ETDEWEB)

Pham, Vuong-Hung [WCU Hybrid Materials Program, Department of Materials Science and Engineering, Seoul National University, Seoul, 151-742 (Korea, Republic of); Advanced Institute for Science and Technology (AIST), Hanoi University of Science and Technology (HUST), No 1, Dai Co Viet Road, Ha Noi (Viet Nam); Lee, Seung-Hee; Li, Yuanlong; Kim, Hyoun-Ee [WCU Hybrid Materials Program, Department of Materials Science and Engineering, Seoul National University, Seoul, 151-742 (Korea, Republic of); Shin, Kwan-Ha [Department of Dental Laboratory Science and Engineering, Korea University, Seoul, 136-703 (Korea, Republic of); Koh, Young-Hag, E-mail: kohyh@korea.ac.kr [Department of Dental Laboratory Science and Engineering, Korea University, Seoul, 136-703 (Korea, Republic of)

2013-06-01

This study reports the utility of tantalum (Ta) coating for improving the surface hardness, in vitro bioactivity and biocompatibility of Co–Cr implants. The use of direct current sputtering allowed for the deposition of a dense and uniform Ta film onto a Co–Cr substrate, which was composed of β-phase Ta grains. This hard Ta coating significantly improved the surface hardness of the Co–Cr by a factor of > 2.3. In addition, the Ta-deposited Co–Cr substrate showed a vigorous precipitation of apatite crystals on its surface after 4 weeks of immersion in simulated body fluid, suggesting its excellent in vitro bioactivity. This bioactive Ta coating led to a considerable improvement in the in vitro biocompatibility of the Co–Cr, which was assessed in terms of the attachment, proliferation and differentiation of pre-osteoblasts (MC3T3-E1). - Highlights: • Dense and uniform Ta film was deposited onto a Co–Cr substrate using DC sputtering. • The Ta coating significantly enhanced the surface hardness of the Co–Cr. • The in vitro biocompatibility of the Co–Cr was also significantly improved.

Utility of tantalum (Ta) coating to improve surface hardness in vitro bioactivity and biocompatibility of Co–Cr

International Nuclear Information System (INIS)

Pham, Vuong-Hung; Lee, Seung-Hee; Li, Yuanlong; Kim, Hyoun-Ee; Shin, Kwan-Ha; Koh, Young-Hag

2013-01-01

This study reports the utility of tantalum (Ta) coating for improving the surface hardness, in vitro bioactivity and biocompatibility of Co–Cr implants. The use of direct current sputtering allowed for the deposition of a dense and uniform Ta film onto a Co–Cr substrate, which was composed of β-phase Ta grains. This hard Ta coating significantly improved the surface hardness of the Co–Cr by a factor of > 2.3. In addition, the Ta-deposited Co–Cr substrate showed a vigorous precipitation of apatite crystals on its surface after 4 weeks of immersion in simulated body fluid, suggesting its excellent in vitro bioactivity. This bioactive Ta coating led to a considerable improvement in the in vitro biocompatibility of the Co–Cr, which was assessed in terms of the attachment, proliferation and differentiation of pre-osteoblasts (MC3T3-E1). - Highlights: • Dense and uniform Ta film was deposited onto a Co–Cr substrate using DC sputtering. • The Ta coating significantly enhanced the surface hardness of the Co–Cr. • The in vitro biocompatibility of the Co–Cr was also significantly improved

Microwave assisted coating of bioactive amorphous magnesium phosphate (AMP) on polyetheretherketone (PEEK).

Science.gov (United States)

Ren, Yufu; Sikder, Prabaha; Lin, Boren; Bhaduri, Sarit B

2018-04-01

Polyetheretherketone (PEEK) with great thermal and chemical stability, desirable mechanical properties and promising biocompatibility is being widely used as orthopedic and dental implant materials. However, the bioinert surface of PEEK can hinder direct osseointegration between the host tissue and PEEK based implants. The important signatures of this paper are as follows. First, we report for the formation of osseointegrable amorphous magnesium phosphate (AMP) coating on PEEK surface using microwave energy. Second, coatings consist of nano-sized AMP particles with a stacked thickness of 800nm. Third, coatings enhance bioactivity in-vitro and induce significantly high amount of bone-like apatite coating, when soaked in simulated body fluid (SBF). Fourth, the as-deposited AMP coatings present no cytotoxicity effects and are beneficial for cell adhesion at early stage. Finally, the high levels of expression of osteocalcin (OCN) in cells cultured on AMP coated PEEK samples indicate that AMP coatings can promote new bone formation and hence osseointegration. Copyright © 2017 Elsevier B.V. All rights reserved.

Effect of phosphate additives on the microstructure, bioactivity, and degradability of microarc oxidation coatings on Mg-Zn-Ca-Mn alloy.

Science.gov (United States)

Dou, Jinhe; You, Qiongya; Gu, Guochao; Chen, Chuanzhong; Zhang, Xihua

2016-09-20

Calcium phosphate coatings were prepared on the surface of self-designed Mg-Zn-Ca-Mn alloy using microarc oxidization technology. To characterize the microstructures, cross-section morphologies, and compositions of the coatings, the authors used scanning electron microscopy equipped with an energy-disperse spectrometer, x-ray diffraction, and Fourier transform infrared spectroscopy. Potentiodynamic polarization in the simulated body fluid (SBF) was used to evaluate the corrosion behaviors of the samples. An SBF immersion test was used to evaluate the coating bioactivity and degradability. After the immersion tests, some bonelike apatite formed on the coating surfaces indicate that bioactivity of the coatings is excellent. The coating prepared in electrolyte containing (NaPO3)6 had slower degradation rate after immersion test for 21 days.

Fabrication of nanoporous Sr incorporated TiO{sub 2} coating on 316L SS: Evaluation of bioactivity and corrosion protection

Energy Technology Data Exchange (ETDEWEB)

Anne Pauline, S. [Department of Chemistry, Anna University, Chennai 600025 (India); Kamachi Mudali, U. [Corrosion Science and Technology Section, IGCAR, Kalpakkam 603102 (India); Rajendran, N., E-mail: nrajendran@annauniv.edu [Department of Chemistry, Anna University, Chennai 600025 (India)

2013-10-01

In this paper, nanoporous TiO{sub 2} and Sr-incorporated TiO{sub 2} coated 316L SS were prepared by sol–gel methodology. The effect of Sr incorporation into TiO{sub 2} coating on bioactivity and corrosion resistance was investigated. Attenuated total reflectance–Fourier transform infrared (ATR–FTIR) spectroscopy, X-ray diffraction analysis (XRD), atomic force microscopy (AFM), scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX) results obtained after in vitro bioactivity test confirm the excellent growth of crystalline hydroxyapatite (HAp) over nanoporous Sr-incorporated TiO{sub 2} coated 316L SS which may be attributed to the slow and steady release of Sr ions from the coatings. The electrochemical evaluation of the coatings confirms that Sr-incorporated TiO{sub 2} coating offer excellent protection to 316L SS by acting as a barrier layer. The results showed that the incorporation of Sr enhanced both bioactivity and corrosion resistance of 316L SS. Hence Sr-incorporated TiO{sub 2} coated 316L SS is a promising material for orthopaedic implant applications. - Highlights: • Nanoporous Sr-incorporated TiO{sub 2} coatings were successfully fabricated on 316L SS. • The coatings have excellent adhesion to the substrate and appreciable Vickers micro hardness value. • Sr-incorporated TiO{sub 2} coated specimens exhibited excellent hydroxyapatite growth due to slow release of Sr from the coating. • Sr incorporation enhances the corrosion resistance of TiO{sub 2} coating.

Muscle-specific integrins in masseter muscle fibers of chimpanzees: an immunohistochemical study.

Directory of Open Access Journals (Sweden)

Gianluigi Vaccarino

2010-05-01

Full Text Available Most notably, recent comparative genomic analyses strongly indicate that the marked differences between modern human and chimpanzees are likely due more to changes in gene regulation than to modifications of the genes. The most peculiar aspect of hominoid karyotypes is that human have 46 chromosomes whereas gorillas and chimpanzees have 48. Interestingly, human and chimpanzees do share identical inversions on chromosome 7 and 9 that are not evident in the gorilla karyotype. Thus, the general phylogeny suggests that humans and chimpanzees are sister taxa; based on this, it seems that human-chimpanzee sequence similarity is an astonishing 99%. At this purpose, of particular interest is the inactivation of the myosin heavy chain 16 (MYH16 gene, most prominently expressed in the masticatory muscle of mammals. It has been showed that the loss of this gene in humans may have resulted in smaller masticatory muscle and consequential changes to cranio-facial morphology and expansion of the human brain case. Powerful masticatory muscles are found in most primates; contrarily, in both modern and fossil member Homo, these muscles are considerably smaller. The evolving hominid masticatory apparatus shifted towards a pattern of gracilization nearly simultaneously with accelerated encephalization in early Homo. To better comprehend the real role of the MYH16 gene, we studied the primary proteins present in the muscle fibers of humans and non-humans, in order to understand if they really can be influenced by MYH16 gene. At this aim we examined the muscle-specific integrins, alpha 7B and beta 1D-integrins, and their relative fetal isoforms, alpha 7A and beta 1A-integrins, analyzing, by immunohistochemistry, muscle biopsies of two components of a chimpanzee's group in captivity, an alpha male and a non-alpha male subjects; all these integrins participate in vital biological processes such as maintenance of tissue integrity, embryonic development, cell

In Situ Laser Coating of Calcium Phosphate on TC4 Surface for Enhancing Bioactivity

Institute of Scientific and Technical Information of China (English)

DENG Chi; WANG Yong; ZHANG Ya-ping; GAO Jia-cheng

2007-01-01

Titanium alloy has been a successful implant material owing to its excellent ratio of strength to weight,toughness, and bio-inert oxide surface. Significant progress has been made in improving the bioactivity of titanium alloy by coating its oxide surface with calcium phosphates. In the present study, in situ coating was reported on Ti6Al4V(TC4) surface with calcium phosphate (Ca-P) bioceramics synthesized and synchronously cladded by laser beam. This coating was grown by first preplacing directly the raw powders, which contain 80% of CaHPO4 ·2H2O, 20% of CaCO3, and dram of rare earth (RE), on the TC4 surfaces, and then exposing the surfaces to the laser beam with a power density of 12. 73-15.27 MW · m-2 and a scanning velocity of 10. 5 m/s. The resultant coating was characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), thermogravimetric analysis and Different-thermal Scanning (TG-DSC), and Energy Dispersive X-ray Detection (EDX). The results show that these laser ceramics include hydroxyapatite (HA), tricalcium phosphate (TCP), Ca2P2O7, and other Ca-P phases, and the interface between the coating and the TC4 substrate has tighter fixation, in which the chemical bonding is approved. These laser hybrid coatings are useful in enhancing the bioactivity of titanium alloy surfaces.

XPS and bioactivity study of the bisphosphonate pamidronate adsorbed onto plasma sprayed hydroxyapatite coatings

International Nuclear Information System (INIS)

McLeod, Kate; Kumar, Sunil; Smart, Roger St.C.; Dutta, Naba; Voelcker, Nicolas H.; Anderson, Gail I.; Sekel, Ron

2006-01-01

This paper reports the use of X-ray photoelectron spectroscopy (XPS) to investigate bisphosphonate (BP) adsorption onto plasma sprayed hydroxyapatite (HA) coatings commonly used for orthopaedic implants. BPs exhibit high binding affinity for the calcium present in HA and hence can be adsorbed onto HA-coated implants to exploit their beneficial properties for improved bone growth at the implant interface. A rigorous XPS analysis of pamidronate, a commonly used nitrogenous BP, adsorbed onto plasma sprayed HA-coated cobalt-chromium substrates has been carried out, aimed at: (a) confirming the adsorption of this BP onto HA; (b) studying the BP diffusion profile in the HA coating by employing the technique of XPS depth profiling; (c) confirming the bioactivity of the adsorbed BP. XPS spectra of plasma sprayed HA-coated discs exposed to a 10 mM aqueous BP solution (pamidronate) for periods of 1, 2 and 24 h showed nitrogen and phosphorous photoelectron signals corresponding to the BP, confirming its adsorption onto the HA substrate. XPS depth profiling of the 2 h BP-exposed HA discs showed penetration of the BP into the HA matrix to depths of at least 260 nm. The bioactivity of the adsorbed BP was confirmed by the observed inhibition of osteoclast (bone resorbing) cell activity. In comparison to the HA sample, the HA sample with adsorbed BP exhibited a 25-fold decrease in primary osteoclast cells

Electrochemical behavior of 45S5 bioactive ceramic coating on Ti6Al4V alloy for dental applications

Science.gov (United States)

Machado López, M. M.; Espitia Cabrera, M. I.; Faure, J.; Contreras García, M. E.

2016-04-01

Titanium and its alloys are widely used as implant materials because of their mechanical properties and non-toxic behavior. Unfortunately, they are not bioinert, which means that they can release ions and can only fix the bone by mechanical anchorage, this can lead to the encapsulation of dense fibrous tissue in the body. The bone fixation is required in clinical conditions treated by orthopedic and dental medicine. The proposal is to coat metallic implants with bioactive materials to establish good interfacial bonds between the metal substrate and bone by increasing bioactivity. Bioactive glasses, ceramics specifically 45 S5 Bioglass, have drawn attention as a serious functional biomaterial because osseointegration capacity. The EPD method of bioglass gel precursor was proposed in the present work as a new method to obtain 45S5/Ti6A14V for dental applications. The coatings, were thermally treated at 700 and 800°C and presented the 45 S5 bioglass characteristic phases showing morphology and uniformity with no defects, quantification percentages by EDS of Si, Ca, Na, P and O elements in the coating scratched powders, showed a good proportional relationship demonstrating the obtention of the 45S5 bioglass. The corrosion tests were carried out in Hank's solution. By Tafel extrapolation, Ti6Al4V alloy showed good corrosion resistance in Hank's solution media, by the formation of a passivation layer on the metal surface, however, in the system 45S5/Ti6Al4V there was an increase in the corrosion resistance; icon-, Ecorr and corrosion rate decreased, the mass loss and the rate of release of ions, were lower in this system than in the titanium alloy without coating.

Influence of PCL on mechanical properties and bioactivity of ZrO{sub 2}-based hybrid coatings synthesized by sol–gel dip coating technique

Energy Technology Data Exchange (ETDEWEB)

Catauro, Michelina, E-mail: michelina.catauro@unina2.it [Department of Industrial and Information Engineering, Second University of Naples, Via Roma 29, 81031 Aversa (Italy); Bollino, Flavia [Department of Industrial and Information Engineering, Second University of Naples, Via Roma 29, 81031 Aversa (Italy); Veronesi, Paolo [Department of Engineering “Enzo Ferrari”, University of Modena and Reggio Emilia, Via Vignolese 905, 41125 Modena (Italy); Lamanna, Giuseppe [Department of Industrial and Information Engineering, Second University of Naples, Via Roma 29, 81031 Aversa (Italy)

2014-06-01

The biological properties of medical implants can be enhanced through surface modifications such as to provide a firm attachment of the implant. In this study, organic–inorganic hybrid coatings have been synthesized via sol–gel dip coating. They consist of an inorganic ZrO{sub 2} matrix in which different amounts of poly(ε-caprolactone) have been entrapped to improve the mechanical properties of the films. The influence of the PCL amount on the microstructural, biological and mechanical properties of the coating has been investigated. Scanning electron microscopy (SEM) and X-ray diffraction (XRD) analyses have shown that the hybrids used for the coating are homogenous and totally amorphous materials; Fourier transform infrared spectroscopy (FT-IR) has demonstrated that hydrogen bonds arise between the organic and inorganic phases. SEM and atomic force microscopy (AFM) have highlighted the nanostructured nature of the film. SEM and EDS analyses, after soaking the samples in a simulated body fluid (SBF), have pointed out the apatite formation on the coating surface, which proves the bone-bonding ability of the nanocomposite bioactive films. Scratch and nano-indentation tests have shown that the coating hardness, stiffness and Young's modulus decrease in the presence of large amounts of the organic phase. - Highlights: • ZrO{sub 2}/PCL organic-inorganic hybrid coatings synthesis via sol-gel dip coating. • Coatings porosity and bioactivity increase in presence of high PCL amount. • Coatings Hardness and Young’s modulus decrease in presence of high PCL amount.

In vitro bioactivity, tribological property, and antibacterial ability of Ca-Si-based coatings doped with cu particles in-situ fabricated by laser cladding

Science.gov (United States)

Hou, Baoping; Yang, Zhao; Yang, Yuling; Zhang, Erlin; Qin, Gaowu

2018-03-01

The present study aimed to in-situ fabricate Ca-Si-based coatings doped with copper particles (Cu-CS coatings) to enhance in vitro bioactivity, tribological property, and antibacterial ability of Ti-6Al-4V alloy. The effects of copper addition on the multiple properties were evaluated. Our results showed that Ca2SiO4, CaTiO3, and Cu2O were in-situ fabricated after laser processing. The Cu-CS coatings exhibited an excellent wear resistance and enhanced wettability. Regarding the in vitro bioactivity, after soaking in simulated body fluid, Cu-CS coatings developed an apatite surface layer that was reduced in the coatings with higher weight percent Cu addition. The Cu-CS coatings enhanced the inhibitory action against E. coli strains, especially for the coating with a higher concentration of Cu in it. Hence, the synthesized Cu-CS coatings present excellent tribological properties, enhanced bioactivity, and antibacterial property, and, therefore, would be used to modify the surface properties of Ti-6Al-4V implants for bone tissue engineering applications.

Bioactive coatings on Portland cement substrates: Surface precipitation of apatite-like crystals

International Nuclear Information System (INIS)

Gallego, Daniel; Higuita, Natalia; Garcia, Felipe; Ferrell, Nicholas; Hansford, Derek J.

2008-01-01

We report a method for depositing bioactive coatings onto cement materials for bone tissue engineering applications. White Portland cement substrates were hydrated under a 20% CO 2 atmosphere, allowing the formation of CaCO 3 . The substrates were incubated in a calcium phosphate solution for 1, 3, and 6 days (CPI, CPII, and CPIII respectively) at 37 deg. C to induce the formation of carbonated apatite. Cement controls were prepared and hydrated with and without CO 2 atmosphere (C+ and C- respectively). The presence of apatite-like crystals was verified by Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS). The substrate cytocompatibility was evaluated via SEM after 24 hour cell cultures. SEM revealed the presence Ca(OH) 2 on C-, and CaCO 3 on C+. Apatite-like crystals were detected only on CPIII, confirmed by phosphorus EDS peaks only for CPIII. Cells attached and proliferated similarly well on all the substrates except C-. These results prove the feasibility of obtaining biocompatible and bioactive coatings on Portland cement for bone tissue engineering applications

Electrophoretic co-deposition of polyvinyl alcohol (PVA) reinforced alginate–Bioglass® composite coating on stainless steel: Mechanical properties and in-vitro bioactivity assessment

International Nuclear Information System (INIS)

Chen, Qiang; Cabanas-Polo, Sandra; Goudouri, Ourania-Menti; Boccaccini, Aldo R.

2014-01-01

PVA reinforced alginate–bioactive glass (BG) composite coatings were produced on stainless steel by a single step electrophoretic deposition (EPD) process. The present paper discusses the co-deposition mechanism of the three components and presents a summary of the relevant properties of the composite coatings deposited from suspensions with different PVA concentrations. Homogeneous composite coatings with compact microstructure and increased thickness, i.e. as high as 10 μm, were observed by scanning electron microscopy (SEM). The surface roughness of coatings with different PVA contents was slightly increased, while a significant increase of water contact angles due to PVA addition was detected and discussed. Improved adhesion strength of coatings containing different amounts of PVA was quantitatively and qualitatively confirmed by pull-off adhesion and cycled bending tests, respectively. In-vitro bioactivity tests were performed in simulated body fluid (SBF) for 0.5, 1, 2, 4, 7, and 14 days, respectively. The decomposition rate of the coatings was reduced with PVA content, and rapid hydroxyapatite forming ability of the composite coatings in SBF was confirmed by FTIR and XRD analyses. According to the results of this study, composite alginate–Bioglass® bioactive coatings combined with PVA are proposed as promising candidates for dental and orthopedic applications. - Highlights: • PVA reinforced alginate–bioactive glass composite coating on stainless steel produced by EPD • The co-deposition mechanism was experimentally confirmed. • Homogeneous and compact coating microstructure obtained by the addition of PVA • Improved adhesion strength of PVA reinforced coatings confirmed qualitatively and quantitatively • Controlled degradation rate and rapid HA forming ability of PVA-containing coatings in SBF

Electrophoretic co-deposition of polyvinyl alcohol (PVA) reinforced alginate–Bioglass® composite coating on stainless steel: Mechanical properties and in-vitro bioactivity assessment

Energy Technology Data Exchange (ETDEWEB)

Chen, Qiang; Cabanas-Polo, Sandra; Goudouri, Ourania-Menti; Boccaccini, Aldo R., E-mail: aldo.boccaccini@ww.uni-erlangen.de

2014-07-01

PVA reinforced alginate–bioactive glass (BG) composite coatings were produced on stainless steel by a single step electrophoretic deposition (EPD) process. The present paper discusses the co-deposition mechanism of the three components and presents a summary of the relevant properties of the composite coatings deposited from suspensions with different PVA concentrations. Homogeneous composite coatings with compact microstructure and increased thickness, i.e. as high as 10 μm, were observed by scanning electron microscopy (SEM). The surface roughness of coatings with different PVA contents was slightly increased, while a significant increase of water contact angles due to PVA addition was detected and discussed. Improved adhesion strength of coatings containing different amounts of PVA was quantitatively and qualitatively confirmed by pull-off adhesion and cycled bending tests, respectively. In-vitro bioactivity tests were performed in simulated body fluid (SBF) for 0.5, 1, 2, 4, 7, and 14 days, respectively. The decomposition rate of the coatings was reduced with PVA content, and rapid hydroxyapatite forming ability of the composite coatings in SBF was confirmed by FTIR and XRD analyses. According to the results of this study, composite alginate–Bioglass® bioactive coatings combined with PVA are proposed as promising candidates for dental and orthopedic applications. - Highlights: • PVA reinforced alginate–bioactive glass composite coating on stainless steel produced by EPD • The co-deposition mechanism was experimentally confirmed. • Homogeneous and compact coating microstructure obtained by the addition of PVA • Improved adhesion strength of PVA reinforced coatings confirmed qualitatively and quantitatively • Controlled degradation rate and rapid HA forming ability of PVA-containing coatings in SBF.

Chitosan-58S bioactive glass nanocomposite coatings on TiO2 nanotube: Structural and biological properties

Science.gov (United States)

Mokhtari, H.; Ghasemi, Z.; Kharaziha, M.; Karimzadeh, F.; Alihosseini, F.

2018-05-01

Bacterial infection and insignificant osseointegration have been recognized as the main reasons of the failures of titanium based implants. The aim of this study was to apply titanium oxide nanotube (TNT) array on titanium using electrochemical anodization process as a more appropriate substrate for chitosan and chitosan-58S bioactive glass (BG) (58S-BG-Chitosan) nanocomposite coatings covered TNTs (TNT/Chiosan, TNT/58S-BG-Chitosan, respectively) through a conventional dip-coating process. Results showed that a TNT layer with average inner diameter of 82 ± 19 nm and wall's thickness of 23 ± 9 nm was developed on titanium surface using electrochemical anodization process. Roughness and contact angle measurement showed that TNT with Ra = 449 nm had highest roughness and hydrophilicity which then reduced to 86 nm and 143 nm for TNT/Chitosan and TNT/58S-BG-Chitosan, respectively. In vitro bioactivity evaluation in simulated buffer fluid (SBF) solution and antibacterial activity assay predicted that TNT/58S-BG-Chitosan was superior in bone like apatite formation and antibacterial activity against both gram-positive and gram-negative bacteria compared to Ti, TNT and TNT/Chitosan samples, respectively. Results revealed the noticeable MG63 cell attachment and proliferation on TNT/58S-BG-Chitosan coating compared to those of uncoated TNTs. These results confirmed the positive effect of using TNT substrate for natural polymer coating on improved bioactivity of implant.

Development of bioactive coatings based on γ-irradiated proteins to preserve strawberries

International Nuclear Information System (INIS)

Vu, K.D.; Hollingsworth, R.G.; Salmieri, S.; Takala, P.N.; Lacroix, M.

2012-01-01

Gamma irradiation was applied for creating cross-linked proteins to enhance the physicochemical properties of edible films made of calcium caseinate, whey protein isolate and glycerol. The characteristics of γ irradiated cross-linked proteins were analyzed by Fourier Transform Infrared spectroscopy. A second derivative spectra exhibited changes in band intensities that were correlated to an increase of β-sheet structure and a decrease of α-helix and unordered fractions of γ irradiated-cross-linked proteins as compared to the control without irradiation. Furthermore, on addition of methylcellulose to the irradiated protein matrix it was found that it has potential in enhancing the puncture strength and has no detrimental effect on water vapor permeability of protein based films. Finally, these film formulations were used as bioactive edible coatings containing natural antimicrobial agents (limonene and peppermint) to preserve the shelf life of fresh strawberries during storage. The bioactive coatings containing peppermint was found to be more efficient as preserving coatings than the formulations containing limonene. Irradiated proteins/methylcellulose/peppermint formulation had only 40% of decay at day 8 while it was 65% for the control. - Highlights: ► Crosslinked proteins and antimicrobials agents was able to preserve strawberries. ► Crosslinked protein structure was more ordered. ► Films based on crosslinked proteins and methylcellulose enhanced puncture strength.

Separation of integrin-dependent adhesion from morphological changes based on differential PLC specificities.

Science.gov (United States)

Wooten, D K; Teague, T K; McIntyre, B W

1999-01-01

In normal lymphocytes an inside-out signal up-regulating integrin adhesion is followed by a ligand-mediated outside-in cell spreading signal. Protein kinase C (PKC) inhibition blocks lymphocyte adherence to and spreading on fibronectin. In contrast, putative PLC inhibitors yield distinct differences with respect to adhesion and morphology. The phosphatidylinositol-specific phospholipase C (PLC) inhibitor neomycin blocked spreading of CD3/CD28-activated T cells on fibronectin by disrupting adhesion. Furthermore, when an additional inside-out signal for fibronectin adhesion is unnecessary such as with HPB-ALL T leukemic or phorbol-myristate-acetate-treated normal T cells, neomycin treatment does not alter adhesion or morphology. However, the phosphatidylcholine-specific PLC inhibitor D609 abrogates cell spreading without affecting adhesion to fibronectin in these cells as well as the CD3/CD28-activated T cells. These results strongly suggest that inside-out signaling for the integrin alpha4beta1 in lymphocytes proceeds through phosphatidylinositol-specific PLC and PKC, whereas the outside-in signal utilizes phosphatidylcholine-specific PLC and PKC.

Effect of Sr on the bioactivity and corrosion resistance of nanoporous niobium oxide coating for orthopaedic applications

International Nuclear Information System (INIS)

Pauline, S. Anne; Rajendran, N.

2014-01-01

In this study, strontium incorporated Nb 2 O 5 was synthesized in two different proportions by sol–gel methodology and was deposited on 316L SS by spin coating method. The synthesis conditions were optimized to obtain a nanoporous morphology. The prepared Sr-incorporated Nb 2 O 5 coatings were uniform, smooth and well adherent on to the substrate 316L SS. The coatings were characterized by attenuated total reflectance-infrared spectroscopy (ATR-IR), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX), atomic force microscopy (AFM) and transmission electron microscopy (TEM) and the formation of Sr-incorporated Nb 2 O 5 coatings with nanoporous morphology was confirmed. Static water contact angle measurements showed an enhancement in the wettability of the obtained coatings. In vitro bioactivity test of the coated substrates showed that 0.05 M Sr-incorporated Nb 2 O 5 coating had better bioactivity compared to 0.1 M Sr-incorporated coating. Solution analysis studies confirmed the controlled release of Sr ions from the coating, which aid and enhance hydroxyapatite (HAp) growth. Electrochemical studies confirmed that the coatings provided excellent corrosion protection to the base material as increased charge transfer resistance and decreased double layer capacitance was observed for the coated substrates. - Highlights: • Nanoporous Sr-incorporated Nb 2 O 5 coatings were deposited on 316L SS. • The coatings have excellent bond strength and high Vickers micro hardness value. • Nanoporous 0.05 M Sr-incorporated Nb 2 O 5 coating showed hydroxyapatite growth. • Slow release of strontium from the coating accelerated hydroxyapatite growth. • The nanoporous coatings offered excellent corrosion protection to 316L SS

β1-integrin controls cell fate specification in early lens development

Science.gov (United States)

Pathania, Mallika; Wang, Yan; Simirskii, Vladimir N.; Duncan, Melinda K.

2016-01-01

Integrins are heterodimeric cell surface molecules that mediate cell-extracellular matrix (ECM) adhesion, ECM assembly, and regulation of both ECM and growth factor induced signaling. However, the developmental context of these diverse functions is not clear. Loss of β1-integrin from the lens vesicle (mouse E10.5) results in abnormal exit of anterior lens epithelial cells (LECs) from the cell cycle and their aberrant elongation toward the presumptive cornea by E12.5. These cells lose expression of LEC markers and initiate expression of the Maf (also known as c-Maf) and Prox1 transcription factors as well as other lens fiber cell markers, β1-integrin null LECs also upregulate the ERK, AKT and Smad1/5/8 phosphorylation indicative of BMP and FGF signaling. By E14.5, β1-integrin null lenses have undergone a complete conversion of all lens epithelial cells into fiber cells. These data suggest that shortly after lens vesicle closure, β1-integrin blocks inappropriate differentiation of the lens epithelium into fibers, potentially by inhibiting BMP and/or FGF receptor activation. Thus, β1-integrin has an important role in fine-tuning the response of the early lens to the gradient of growth factors that regulate lens fiber cell differentiation. PMID:27596755

Development and preclinical evaluation of radiolabelled somatostatin receptor agonists and αvβ3-integrin antagonists

International Nuclear Information System (INIS)

Stoecklin, G.; Wester, H.J.; Haubner, R.; Schottelius, M.

2002-01-01

Tumours express specific receptors for peptide ligands. This can be exploited for tumour targeting. New bioactive peptides are available, in particular new somatostatin analogs and RGD-Peptides for targeting the α V β 3 -integrin. The design and optimization of radiolabelled peptides with respect to their receptor affinity, tumour uptake, biodistribution, pharmacokinetics and stability may provide better tracers for tumour imaging and therapy. Based on two basic structures, new radioiodinated and carbohydrated somatostatin analogs and cyclic RGD-peptides were developed and evaluated. (author)

The in vitro indirect cytotoxicity test and in vivo interface bioactivity evaluation of biodegradable FHA coated Mg-Zn alloys

Energy Technology Data Exchange (ETDEWEB)

Li Jianan [State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240 (China); Han Pei, E-mail: hanpei_cn@163.com [Orthopaedic Department of the 6th People' s Hospital, Shanghai Jiao Tong University, Shanghai 200233 (China); Ji Weiping [Orthopaedic Department of the 6th People' s Hospital, Shanghai Jiao Tong University, Shanghai 200233 (China); Song, Yang; Zhang, Shaoxiang; Chen Ying; Zhao Changli; Zhang Fan [State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240 (China); Zhang Xiaonong, E-mail: xnzhang@sjtu.edu.cn [State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240 (China); Key Laboratory of Inorganic Coating Materials, Chinese Academy of Sciences, Shanghai 200051 (China); Jiang Yao [Orthopaedic Department of the 6th People' s Hospital, Shanghai Jiao Tong University, Shanghai 200233 (China)

2011-12-15

A kind of biodegradable fluoridated hydroxyapatite (FHA) coating was prepared on Mg-Zn alloy to improve the interface bioactivity in bone healing via electrodeposition method. The in vitro cytotoxicity evaluation of the ions released during degradation was taken. No toxicity was shown and even higher cells' viability appeared on the 7th day compared with the normal culture case (negative control). In vivo implantation was carried out in the femoral condyle of adult New Zealand rabbits. The cross section showed by Micro-CT scan confirmed that the better interface contacts happened in the coated group after one month implantation. Also the coating left can still be normally observed by scanning electron microscope (SEM) with a little degradation. As a result, the FHA coating may be a promising candidate to enhance interface bioactivity for biodegradable Mg alloys in orthopaedics.

Osseointegration properties of titanium dental implants modified with a nanostructured coating based on ordered porous silica and bioactive glass nanoparticles

Science.gov (United States)

Covarrubias, Cristian; Mattmann, Matías; Von Marttens, Alfredo; Caviedes, Pablo; Arriagada, Cristián; Valenzuela, Francisco; Rodríguez, Juan Pablo; Corral, Camila

2016-02-01

The fabrication of a nanoporous silica coating loaded with bioactive glass nanoparticles (nBG/NSC) on titanium dental implant surface and its in vitro and in vivo evaluation is presented. The coating was produced by a combined sol-gel and evaporation induced self-assembly process. In vitro bioactivity was assessed in simulated body fluid (SBF) and investigating the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). A rat tibial model was employed to analyze the bone response to nBG/NSC-modified titanium implant surface in vivo. The nBG/NSC coating was confirmed at nano level to be constituted by a highly ordered nanoporous silica structure. The coating nanotopography in conjunction with the bioactivity of the BG particles accelerate the in vitro apatite formation and promote the osteogenic differentiation of hBMSCs in absence of osteogenic supplements. These properties accelerate the formation of bone tissue in the periphery of the implant after 3 weeks of implantation. Backscattered scanning electron microscopy images revealed the presence of gaps and soft tissue in the unmodified implant after 6 weeks, whereas the nBG/NSC-modified implant showed mature bone in intimate contact with the implant surface. The nBG/NSC coating appears promising for accelerating the osseointegration of dental implants.

Effect of Sr on the bioactivity and corrosion resistance of nanoporous niobium oxide coating for orthopaedic applications

Energy Technology Data Exchange (ETDEWEB)

Pauline, S. Anne; Rajendran, N., E-mail: nrajendran@annauniv.edu

2014-03-01

In this study, strontium incorporated Nb{sub 2}O{sub 5} was synthesized in two different proportions by sol–gel methodology and was deposited on 316L SS by spin coating method. The synthesis conditions were optimized to obtain a nanoporous morphology. The prepared Sr-incorporated Nb{sub 2}O{sub 5} coatings were uniform, smooth and well adherent on to the substrate 316L SS. The coatings were characterized by attenuated total reflectance-infrared spectroscopy (ATR-IR), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX), atomic force microscopy (AFM) and transmission electron microscopy (TEM) and the formation of Sr-incorporated Nb{sub 2}O{sub 5} coatings with nanoporous morphology was confirmed. Static water contact angle measurements showed an enhancement in the wettability of the obtained coatings. In vitro bioactivity test of the coated substrates showed that 0.05 M Sr-incorporated Nb{sub 2}O{sub 5} coating had better bioactivity compared to 0.1 M Sr-incorporated coating. Solution analysis studies confirmed the controlled release of Sr ions from the coating, which aid and enhance hydroxyapatite (HAp) growth. Electrochemical studies confirmed that the coatings provided excellent corrosion protection to the base material as increased charge transfer resistance and decreased double layer capacitance was observed for the coated substrates. - Highlights: • Nanoporous Sr-incorporated Nb{sub 2}O{sub 5} coatings were deposited on 316L SS. • The coatings have excellent bond strength and high Vickers micro hardness value. • Nanoporous 0.05 M Sr-incorporated Nb{sub 2}O{sub 5} coating showed hydroxyapatite growth. • Slow release of strontium from the coating accelerated hydroxyapatite growth. • The nanoporous coatings offered excellent corrosion protection to 316L SS.

Tumour exosome integrins determine organotropic metastasis.

Science.gov (United States)

Hoshino, Ayuko; Costa-Silva, Bruno; Shen, Tang-Long; Rodrigues, Goncalo; Hashimoto, Ayako; Tesic Mark, Milica; Molina, Henrik; Kohsaka, Shinji; Di Giannatale, Angela; Ceder, Sophia; Singh, Swarnima; Williams, Caitlin; Soplop, Nadine; Uryu, Kunihiro; Pharmer, Lindsay; King, Tari; Bojmar, Linda; Davies, Alexander E; Ararso, Yonathan; Zhang, Tuo; Zhang, Haiying; Hernandez, Jonathan; Weiss, Joshua M; Dumont-Cole, Vanessa D; Kramer, Kimberly; Wexler, Leonard H; Narendran, Aru; Schwartz, Gary K; Healey, John H; Sandstrom, Per; Labori, Knut Jørgen; Kure, Elin H; Grandgenett, Paul M; Hollingsworth, Michael A; de Sousa, Maria; Kaur, Sukhwinder; Jain, Maneesh; Mallya, Kavita; Batra, Surinder K; Jarnagin, William R; Brady, Mary S; Fodstad, Oystein; Muller, Volkmar; Pantel, Klaus; Minn, Andy J; Bissell, Mina J; Garcia, Benjamin A; Kang, Yibin; Rajasekhar, Vinagolu K; Ghajar, Cyrus M; Matei, Irina; Peinado, Hector; Bromberg, Jacqueline; Lyden, David

2015-11-19

Ever since Stephen Paget's 1889 hypothesis, metastatic organotropism has remained one of cancer's greatest mysteries. Here we demonstrate that exosomes from mouse and human lung-, liver- and brain-tropic tumour cells fuse preferentially with resident cells at their predicted destination, namely lung fibroblasts and epithelial cells, liver Kupffer cells and brain endothelial cells. We show that tumour-derived exosomes uptaken by organ-specific cells prepare the pre-metastatic niche. Treatment with exosomes from lung-tropic models redirected the metastasis of bone-tropic tumour cells. Exosome proteomics revealed distinct integrin expression patterns, in which the exosomal integrins α6β4 and α6β1 were associated with lung metastasis, while exosomal integrin αvβ5 was linked to liver metastasis. Targeting the integrins α6β4 and αvβ5 decreased exosome uptake, as well as lung and liver metastasis, respectively. We demonstrate that exosome integrin uptake by resident cells activates Src phosphorylation and pro-inflammatory S100 gene expression. Finally, our clinical data indicate that exosomal integrins could be used to predict organ-specific metastasis.

Novel Bioactive Antimicrobial Lignin Containing Coatings on Titanium Obtained by Electrophoretic Deposition

Directory of Open Access Journals (Sweden)

Sanja Erakovic

2014-07-01

Full Text Available Hydroxyapatite (HAP is the most suitable biocompatible material for bone implant coatings; its brittleness, however, is a major obstacle, and the reason why research focuses on creating composites with biopolymers. Organosolv lignin (Lig is used for the production of composite coatings, and these composites were examined in this study. Titanium substrate is a key biomedical material due to its well-known properties, but infections of the implantation site still impose a serious threat. One approach to prevent infection is to improve antimicrobial properties of the coating material. Silver doped hydroxyapatite (Ag/HAP and HAP coatings on titanium were obtained by an electrophoretic deposition method in order to control deposited coating mass and morphology by varying applied voltage and deposition time. The effect of lignin on microstructure, morphology and thermal behavior of biocomposite coatings was investigated. The results showed that higher lignin concentrations protect the HAP lattice during sintering, improving coating stability. The corrosion stability was evaluated in simulated body fluid (SBF at 37 °C. Newly formed plate-shaped carbonate-HAP was detected, indicating enhanced bioactive performance. The antimicrobial efficiency of Ag/HAP/Lig was confirmed by its higher reduction of bacteria Staphylococcus aureus TL (S. aureus TL than of HAP/Lig coating. Cytotoxicity assay revealed that both coatings can be classified as non-toxic against healthy immunocompetent peripheral blood mononuclear cells (PBMC.

Electrophoretic co-deposition of polyvinyl alcohol (PVA) reinforced alginate-Bioglass® composite coating on stainless steel: mechanical properties and in-vitro bioactivity assessment.

Science.gov (United States)

Chen, Qiang; Cabanas-Polo, Sandra; Goudouri, Ourania-Menti; Boccaccini, Aldo R

2014-07-01

PVA reinforced alginate-bioactive glass (BG) composite coatings were produced on stainless steel by a single step electrophoretic deposition (EPD) process. The present paper discusses the co-deposition mechanism of the three components and presents a summary of the relevant properties of the composite coatings deposited from suspensions with different PVA concentrations. Homogeneous composite coatings with compact microstructure and increased thickness, i.e. as high as 10 μm, were observed by scanning electron microscopy (SEM). The surface roughness of coatings with different PVA contents was slightly increased, while a significant increase of water contact angles due to PVA addition was detected and discussed. Improved adhesion strength of coatings containing different amounts of PVA was quantitatively and qualitatively confirmed by pull-off adhesion and cycled bending tests, respectively. In-vitro bioactivity tests were performed in simulated body fluid (SBF) for 0.5, 1, 2, 4, 7, and 14 days, respectively. The decomposition rate of the coatings was reduced with PVA content, and rapid hydroxyapatite forming ability of the composite coatings in SBF was confirmed by FTIR and XRD analyses. According to the results of this study, composite alginate-Bioglass® bioactive coatings combined with PVA are proposed as promising candidates for dental and orthopedic applications. Copyright © 2014 Elsevier B.V. All rights reserved.

Corrosion and bioactivity performance of graphene oxide coating on Ti−Nb shape memory alloys in simulated body fluid

Energy Technology Data Exchange (ETDEWEB)

Saud, Safaa N., E-mail: safaaengineer@gmail.com [Faculty of Mechanical Engineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Faculty of Information Science and Engineering, Management and Science University, 40100 Shah Alam (Malaysia); Hosseinian, Raheleh S. [Department of Physics, University of Malaya, 50603 Kuala Lumpur (Malaysia); Bakhsheshi-Rad, H.R. [Faculty of Mechanical Engineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Advanced Materials Research Center, Faculty of Materials Engineering, NajafAbad Branch, Islamic Azad University, NajafAbad (Iran, Islamic Republic of); Yaghoubidoust, F. [Ibnu Sina Institute for Fundamental Science Studies, Universiti Teknologi Malaysia, 81310, UTM, Skudai, Johor (Malaysia); Iqbal, N. [Medical Devices & Technology Group (MEDITEG), Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Hamzah, E. [Faculty of Mechanical Engineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Ooi, C.H. Raymond [Department of Physics, University of Malaya, 50603 Kuala Lumpur (Malaysia)

2016-11-01

In the present work, the microstructure, corrosion, and bioactivity of graphene oxide (GO) coating on the laser-modified and -unmodified surfaces of Ti−Nb shape memory alloys (SMAs) were investigated. The surface morphology and chemical composition was examined using field emission scanning electron microscopy (FE-SEM) and X-ray diffraction (XRD). The surface modification was carried out via a femtosecond laser with the aim to increase the surface roughness, and thus increase the adhesion property. FE-SEM analysis of the laser-treated Ti-30 at.% Nb revealed the increase in surface roughness and oxygen/nitrogen containing groups on the Ti-30 at.% Nb surface after being surface modified via a femtosecond laser. Furthermore, the thickness of GO was increased from 35 μm to 45 μm after the surface was modified. Potentiodynamic polarisation and electrochemical impedance spectroscopy studies revealed that both the GO and laser/GO-coated samples exhibited higher corrosion resistance than that of the uncoated Ti−Nb SMA sample. However, the laser/GO-coated sample presented the highest corrosion resistance in SBF at 37 °C. In addition, during soaking in the simulated body fluid (SBF), both the GO and laser/GO coating improved the formation of apatite layer. Based on the bioactivity results, the GO coating exhibited a remarkable antibacterial activity against gram-negative bacteria compared with the uncoated. In conclusion, the present results indicate that Ti-30 at.% Nb SMAs may be promising alternatives to NiTi for certain biomedical applications. - Highlights: • Ti-30 at.% Nb SMA were successfully produced by PM and microwave sintering. • GO were successfully coated on the surface of TiNb. • The corrosion resistance of TiNb have enhanced after surface modification and coating of GO. • The corrosion resistance increased from 620.7 to 1,760.7 Ω cm{sup 2} with coating of GO. • The GO and laser/GO coating induce apatite formation and enhance the bioactivity.

Corrosion and bioactivity performance of graphene oxide coating on Ti−Nb shape memory alloys in simulated body fluid

International Nuclear Information System (INIS)

Saud, Safaa N.; Hosseinian, Raheleh S.; Bakhsheshi-Rad, H.R.; Yaghoubidoust, F.; Iqbal, N.; Hamzah, E.; Ooi, C.H. Raymond

2016-01-01

In the present work, the microstructure, corrosion, and bioactivity of graphene oxide (GO) coating on the laser-modified and -unmodified surfaces of Ti−Nb shape memory alloys (SMAs) were investigated. The surface morphology and chemical composition was examined using field emission scanning electron microscopy (FE-SEM) and X-ray diffraction (XRD). The surface modification was carried out via a femtosecond laser with the aim to increase the surface roughness, and thus increase the adhesion property. FE-SEM analysis of the laser-treated Ti-30 at.% Nb revealed the increase in surface roughness and oxygen/nitrogen containing groups on the Ti-30 at.% Nb surface after being surface modified via a femtosecond laser. Furthermore, the thickness of GO was increased from 35 μm to 45 μm after the surface was modified. Potentiodynamic polarisation and electrochemical impedance spectroscopy studies revealed that both the GO and laser/GO-coated samples exhibited higher corrosion resistance than that of the uncoated Ti−Nb SMA sample. However, the laser/GO-coated sample presented the highest corrosion resistance in SBF at 37 °C. In addition, during soaking in the simulated body fluid (SBF), both the GO and laser/GO coating improved the formation of apatite layer. Based on the bioactivity results, the GO coating exhibited a remarkable antibacterial activity against gram-negative bacteria compared with the uncoated. In conclusion, the present results indicate that Ti-30 at.% Nb SMAs may be promising alternatives to NiTi for certain biomedical applications. - Highlights: • Ti-30 at.% Nb SMA were successfully produced by PM and microwave sintering. • GO were successfully coated on the surface of TiNb. • The corrosion resistance of TiNb have enhanced after surface modification and coating of GO. • The corrosion resistance increased from 620.7 to 1,760.7 Ω cm 2 with coating of GO. • The GO and laser/GO coating induce apatite formation and enhance the bioactivity

Bioactivity response of Ta_1_-_xO_x coatings deposited by reactive DC magnetron sputtering

International Nuclear Information System (INIS)

Almeida Alves, C.F.; Cavaleiro, A.; Carvalho, S.

2016-01-01

The use of dental implants is sometimes accompanied by failure due to periimplantitis disease and subsequently poor esthetics when soft–hard tissue margin recedes. As a consequence, further research is needed for developing new bioactive surfaces able to enhance the osseous growth. Tantalum (Ta) is a promising material for dental implants since, comparing with titanium (Ti), it is bioactive and has an interesting chemistry which promotes the osseointegration. Another promising approach for implantology is the development of implants with oxidized surfaces since bone progenitor cells interact with the oxide layer forming a diffusion zone due to its ability to bind with calcium which promotes a stronger bond. In the present report Ta-based coatings were deposited by reactive DC magnetron sputtering onto Ti CP substrates in an Ar + O_2 atmosphere. In order to assess the osteoconductive response of the studied materials, contact angle and in vitro tests of the samples immersed in Simulated Body Fluid (SBF) were performed. Structural results showed that oxide phases where achieved with larger amounts of oxygen (70 at.% O). More compact and smooth coatings were deposited by increasing the oxygen content. The as-deposited Ta coating presented the most hydrophobic character (100°); with increasing oxygen amount contact angles progressively diminished, down to the lowest measured value, 63°. The higher wettability is also accompanied by an increase on the surface energy. Bioactivity tests demonstrated that highest O-content coating, in good agreement with wettability and surface energy values, showed an increased affinity for apatite adhesion, with higher Ca/P ratio formation, when compared to the bare Ti substrates. - Highlights: • Ta_1_-_xO_x coatings were deposited by reactive DC magnetron sputtering. • Amorphous oxide phases were achieved with higher oxygen amounts. • Contact angles progressively diminished, with increasing oxygen content. • Ta oxide surface

α6-Integrin alternative splicing: distinct cytoplasmic variants in stem cell fate specification and niche interaction.

Science.gov (United States)

Zhou, Zijing; Qu, Jing; He, Li; Peng, Hong; Chen, Ping; Zhou, Yong

2018-05-02

α6-Integrin subunit (also known as CD49f) is a stemness signature that has been found on the plasma membrane of more than 30 stem cell populations. A growing body of studies have focused on the critical role of α6-containing integrins (α6β1 and α6β4) in the regulation of stem cell properties, lineage-specific differentiation, and niche interaction. α6-Integrin subunit can be alternatively spliced at the post-transcriptional level, giving rise to divergent isoforms which differ in the cytoplasmic and/or extracellular domains. The cytoplasmic domain of integrins is an important functional part of integrin-mediated signals. Structural changes in the cytoplasmic domain of α6 provide an efficient means for the regulation of stem cell responses to biochemical stimuli and/or biophysical cues in the stem cell niche, thus impacting stem cell fate determination. In this review, we summarize the current knowledge on the structural variants of the α6-integrin subunit and spatiotemporal expression of α6 cytoplasmic variants in embryonic and adult stem/progenitor cells. We highlight the roles of α6 cytoplasmic variants in stem cell fate decision and niche interaction, and discuss the potential mechanisms involved. Understanding of the distinct functions of α6 splicing variants in stem cell biology may inform the rational design of novel stem cell-based therapies for a range of human diseases.

Electrophoretic Deposition as a New Bioactive Glass Coating Process for Orthodontic Stainless Steel

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Kyotaro Kawaguchi

2017-11-01

Full Text Available This study investigated the surface modification of orthodontic stainless steel using electrophoretic deposition (EPD of bioactive glass (BG. The BG coatings were characterized by spectrophotometry, scanning electron microscopy with energy dispersive X-ray spectrometry, and X-ray diffraction. The frictional properties were investigated using a progressive load scratch test. The remineralization ability of the etched dental enamel was studied according to the time-dependent mechanical properties of the enamel using a nano-indentation test. The EPD process using alternating current produced higher values in both reflectance and lightness. Additionally, the BG coating was thinner than that prepared using direct current, and was completely amorphous. All of the BG coatings displayed good interfacial adhesion, and Si and O were the major components. Most BG-coated specimens produced slightly higher frictional forces compared with non-coated specimens. The hardness and elastic modulus of etched enamel specimens immersed with most BG-coated specimens recovered significantly with increasing immersion time compared with the non-coated specimen, and significant acid-neutralization was observed for the BG-coated specimens. The surface modification technique using EPD and BG coating on orthodontic stainless steel may assist the development of new non-cytotoxic orthodontic metallic appliances having satisfactory appearance and remineralization ability.

Investigation of bioactivity and cell effects of nano-porous sol–gel derived bioactive glass film

Energy Technology Data Exchange (ETDEWEB)

Ma, Zhijun, E-mail: mokuu@zju.edu.cn [State Key Laboratory of Luminescent Materials and Devices, South China University of Technology, Guangzhou, 510640 (China); Ji, Huijiao [College of Life Science, Zhejiang University, Hangzhou, 310028 (China); Hu, Xiaomeng [School of Materials Science and Engineering, South China University of Technology, Guangzhou, 510640 (China); Teng, Yu [State Key Laboratory of Luminescent Materials and Devices, South China University of Technology, Guangzhou, 510640 (China); Zhao, Guiyun; Mo, Lijuan; Zhao, Xiaoli [College of Life Science, Zhejiang University, Hangzhou, 310028 (China); Chen, Weibo [School of Materials Science and Engineering, South China University of Technology, Guangzhou, 510640 (China); Qiu, Jianrong, E-mail: qjr@scut.edu.cn [State Key Laboratory of Luminescent Materials and Devices, South China University of Technology, Guangzhou, 510640 (China); Zhang, Ming, E-mail: zhangming201201@126.com [College of Life Science, Zhejiang University, Hangzhou, 310028 (China)

2013-11-01

In orthopedic surgery, bioactive glass film coating is extensively studied to improve the synthetic performance of orthopedic implants. A lot of investigations have confirmed that nano-porous structure in bioactive glasses can remarkably improve their bioactivity. Nevertheless, researches on preparation of nano-porous bioactive glasses in the form of film coating and their cell response activities are scarce. Herein, we report the preparation of nano-porous bioactive glass film on commercial glass slide based on a sol–gel technique, together with the evaluation of its in vitro bioactivity through immersion in simulated body fluid and monitoring the precipitation of apatite-like layer. Cell responses of the samples, including attachment, proliferation and osteogenic differentiation, were also investigated using BMSCS (bone marrow derived mesenchymal stem cells) as a model. The results presented here provide some basic information on structural influence of bioactive glass film on the improvement of bioactivity and cellular effects.

Investigation of bioactivity and cell effects of nano-porous sol-gel derived bioactive glass film

Science.gov (United States)

Ma, Zhijun; Ji, Huijiao; Hu, Xiaomeng; Teng, Yu; Zhao, Guiyun; Mo, Lijuan; Zhao, Xiaoli; Chen, Weibo; Qiu, Jianrong; Zhang, Ming

2013-11-01

In orthopedic surgery, bioactive glass film coating is extensively studied to improve the synthetic performance of orthopedic implants. A lot of investigations have confirmed that nano-porous structure in bioactive glasses can remarkably improve their bioactivity. Nevertheless, researches on preparation of nano-porous bioactive glasses in the form of film coating and their cell response activities are scarce. Herein, we report the preparation of nano-porous bioactive glass film on commercial glass slide based on a sol-gel technique, together with the evaluation of its in vitro bioactivity through immersion in simulated body fluid and monitoring the precipitation of apatite-like layer. Cell responses of the samples, including attachment, proliferation and osteogenic differentiation, were also investigated using BMSCS (bone marrow derived mesenchymal stem cells) as a model. The results presented here provide some basic information on structural influence of bioactive glass film on the improvement of bioactivity and cellular effects.

Bioactivity and osteointegration of hydroxyapatite-coated stainless steel and titanium wires used for intramedullary osteosynthesis.

Science.gov (United States)

Popkov, Arnold V; Gorbach, Elena N; Kononovich, Natalia A; Popkov, Dmitry A; Tverdokhlebov, Sergey I; Shesterikov, Evgeniy V

2017-08-01

A lot of research was conducted on the use of various biomaterials in orthopedic surgery. Our study investigated the effects of nanostructured calcium-phosphate coating on metallic implants introduced into the bone marrow canal. Stainless steel or titanium 2-mm wires (groups 1 and 2, respectively), and hydroxyapatite-coated stainless steel or titanium wires of the same diameter (groups 3 and 4, respectively) were introduced into the tibial bone marrow canal of 20 dogs (each group = 5 dogs). Hydroxyapatite coating was deposited on the wires with the method of microarc oxidation. Light microscopy to study histological diaphyseal transverse sections, scanning electron microscopy to study the bone marrow area around the implant and an X-ray electron probe analyzer to study the content of calcium and phosphorus were used to investigate bioactivity and osteointegration after a four weeks period. Osteointegration was also assessed by measuring wires' pull-off strength with a sensor dynamometer. Bone formation was observed round the wires in the bone marrow canal in all the groups. Its intensity depended upon the features of wire surfaces and implant materials. Maximum percentage volume of trabecular bone was present in the bone marrow canals of group 4 dogs that corresponded to a mean of 27.1 ± 0.14%, while it was only 6.7% in group 1. The coating in groups 3 and 4 provided better bioactivity and osteointegration. Hydroxyapatite-coated titanium wires showed the highest degree of bone formation around them and greater pull-off strength. Nanostructured hydroxyapatite coating of metallic wires induces an expressed bone formation and provides osteointegration. Hydroxyapatite-coated wires could be used along with external fixation for bone repair enhancement in diaphyseal fractures, management of osteogenesis imperfecta and correction of bone deformities in phosphate diabetes.

Antimicrobial coatings based on zinc oxide and orange oil for improved bioactive wound dressings and other applications.

Science.gov (United States)

Rădulescu, Marius; Andronescu, Ecaterina; Cirja, Andreea; Holban, Alina Maria; Mogoantă, LaurenŢiu; Bălşeanu, Tudor Adrian; Cătălin, Bogdan; Neagu, Tiberiu Paul; Lascăr, Ioan; Florea, Denisa Alexandra; Grumezescu, Alexandru Mihai; Ciubuca, Bianca; Lazăr, Veronica; Chifiriuc, Mariana Carmen; Bolocan, Alexandra

2016-01-01

This work presents a novel nano-modified coating for wound dressings and other medical devices with anti-infective properties, based on functionalized zinc oxide nanostructures and orange oil (ZnO@OO). The obtained nanosurfaces were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), selected area electron diffraction (SAED), differential thermal analysis-thermogravimetry (DTA-TG), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy. The obtained nanocomposite coatings exhibited an antimicrobial activity superior to bare ZnO nanoparticles (NPs) and to the control antibiotic against Staphylococcus aureus and Escherichia coli, as revealed by the lower minimal inhibitory concentration values. For the quantitative measurement of biofilm-embedded microbial cells, a culture-based, viable cell count method was used. The coated wound dressings proved to be more resistant to S. aureus microbial colonization and biofilm formation compared to the uncoated controls. These results, correlated with the good in vivo biodistribution open new directions for the design of nanostructured bioactive coating and surfaces, which can find applications in the medical field, for obtaining improved bioactive wound dressings and prosthetic devices, but also in food packaging and cosmetic industry.

Characterization of new bioactive coatings of hydroxyapatite and TiO2 obtained by High-Velocity Oxy-Fuel

International Nuclear Information System (INIS)

Melero, H.; Fernandez, J.; Dosta, S.; Guilemany, J. M.

2011-01-01

Hydroxyapatite (Hap: Ca 1 0(PO 4 ) 6 OH 2 ) is a biocompatible and bioactive ceramic material widely used as a coating on metal surfaces (dental implants, hip replacements ...), but the low adhesion between Hap and the substrate, due to differences in thermal expansion coefficients of both (very important in thermal spraying because of the fast cooling of the coating, which can produce a lost of adherence), and the degradation of Hap, have been tried to be improved through the incorporation of TiO 2 to get a good combination of mechanical properties. Therefore, the objective of this project is to produce coatings of Hap 80% TiO 2 and 20% (by weight) on Ti6Al4V by High-Speed Thermal Spray (HVOF). The study of the microstructure has been carried out using scanning electron microscopy and characterization of the crystalline phases by X-ray diffraction and Raman spectrometry. The coatings adhesion has been measured by tensile tests according to ASTM C633-01 (2008), and their bioactivity also has been evaluated through its immersion in simulated body fluid (SBF), in order to measure their capacity to form an apatite layer on their surface. (Author) 26 refs.

Echicetin coated polystyrene beads: a novel tool to investigate GPIb-specific platelet activation and aggregation.

Directory of Open Access Journals (Sweden)

Alexey Navdaev

Full Text Available von Willebrand factor/ristocetin (vWF/R induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling. Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets, while under the same conditions vWF/R treatment led only to αIIbβ3-independent platelet agglutination. The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation, while the total amount of echicetin used for activation is not critical. Echicetin beads induced strong phosphorylation of several proteins including p38, ERK and PKB. Synergistic signaling via P2Y12 and thromboxane receptor through secreted ADP and TxA2, respectively, were important for echicetin bead triggered platelet activation. Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation. Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways.

Echicetin Coated Polystyrene Beads: A Novel Tool to Investigate GPIb-Specific Platelet Activation and Aggregation

Science.gov (United States)

Petunin, Alexey; Clemetson, Kenneth J.; Gambaryan, Stepan; Walter, Ulrich

2014-01-01

von Willebrand factor/ristocetin (vWF/R) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling. Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets, while under the same conditions vWF/R treatment led only to αIIbβ3-independent platelet agglutination. The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation, while the total amount of echicetin used for activation is not critical. Echicetin beads induced strong phosphorylation of several proteins including p38, ERK and PKB. Synergistic signaling via P2Y12 and thromboxane receptor through secreted ADP and TxA2, respectively, were important for echicetin bead triggered platelet activation. Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation. Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways. PMID:24705415

Electrophoretic Deposition as a New Bioactive Glass Coating Process for Orthodontic Stainless Steel

OpenAIRE

Kyotaro Kawaguchi; Masahiro Iijima; Kazuhiko Endo; Itaru Mizoguchi

2017-01-01

This study investigated the surface modification of orthodontic stainless steel using electrophoretic deposition (EPD) of bioactive glass (BG). The BG coatings were characterized by spectrophotometry, scanning electron microscopy with energy dispersive X-ray spectrometry, and X-ray diffraction. The frictional properties were investigated using a progressive load scratch test. The remineralization ability of the etched dental enamel was studied according to the time-dependent mechanical proper...

Coatings of titanium substrates with xCaO·(1 − x)SiO{sub 2} sol–gel materials: characterization, bioactivity and biocompatibility evaluation

Energy Technology Data Exchange (ETDEWEB)

Catauro, M., E-mail: michelina.catauro@unina2.it; Papale, F.; Bollino, F.

2016-01-01

The objective of this study has been to develop low temperature sol–gel coatings to modify the surface of commercially pure titanium grade 4 (a material generally used in dental application) and to evaluate their bioactivity and biocompatibility on the substrate. Glasses of composition expressed by the following general formula xCaO·(1 − x)SiO{sub 2} (0.0 < x < 0.60) have been prepared by means of the sol–gel route starting from tetraethyl orthosilicate and calcium nitrate tetrahydrate. Those materials, still in the sol phase, have been used to coat titanium substrates by means of the dip-coating technique. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) allowed the materials to be characterized and a microstructural analysis of the coatings obtained was performed using scanning electron microscopy (SEM). The potential applications of the coatings in the biomedical field were evaluated by bioactivity and biocompatibility tests. The coated titanium was immersed in simulated body fluid (SBF) for 21 days and the hydroxyapatite deposition on its surface was subsequently evaluated via SEM–EDXS analysis, as an index of bone-bonding capability. To investigate cell-material interactions, mouse embryonic fibroblast cells (3 T3) were seeded onto the specimens and the cell viability was evaluated by a WST-8 assay. - Highlights: • CaO/SiO{sub 2} biomaterials synthesized by sol–gel method at various molar ratio • Coating of titanium substrate with dip-coating technology • Chemical and morphological characterization of materials and coating • Biocompatibility and bioactivity improvement of coated titanium.

Porous SiO_2 nanofiber grafted novel bioactive glass–ceramic coating: A structural scaffold for uniform apatite precipitation and oriented cell proliferation on inert implant

International Nuclear Information System (INIS)

Das, Indranee; De, Goutam; Hupa, Leena; Vallittu, Pekka K.

2016-01-01

A composite bioactive glass–ceramic coating grafted with porous silica nanofibers was fabricated on inert glass to provide a structural scaffold favoring uniform apatite precipitation and oriented cell proliferation. The coating surfaces were investigated thoroughly before and after immersion in simulated body fluid. In addition, the proliferation behavior of fibroblast cells on the surface was observed for several culture times. The nanofibrous exterior of this composite bioactive coating facilitated homogeneous growth of flake-like carbonated hydroxyapatite layer within a short period of immersion. Moreover, the embedded porous silica nanofibers enhanced hydrophilicity which is required for proper cell adhesion on the surface. The cells proliferated well following a particular orientation on the entire coating by the assistance of nanofibrous scaffold-like structural matrix. This newly engineered composite coating was effective in creating a biological structural matrix favorable for homogeneous precipitation of calcium phosphate, and organized cell growth on the inert glass surface. - Highlights: • Fabricated porous SiO_2 nanofibers grafted composite bioactive glass–ceramic coating on inert glass. • The newly engineered coating facilitates uniformly dense apatite precipitation. • Embedded porous silica nanofibers enhance hydrophilicity of the coated surface. • Cells proliferate well on the entire coating following a particular orientation by the assistance of nanofibers. • The coatings have potential to be used as biological scaffold on the surface of implants.

Influence of bioactive material coating of Ti dental implant surfaces on early healing and osseointegration of bone

International Nuclear Information System (INIS)

Yeo, In-Sung; Min, Seung-Ki; An, Young-Bai

2010-01-01

The dental implant surface type is one of many factors that determine the long-term clinical success of implant restoration. The implant surface consists of bioinert titanium oxide, but recently coatings with bioactive calcium phosphate ceramics have often been used on Ti implant surfaces. Bio-active surfaces are known to significantly improve the healing time of the human bone around the inserted dental implant. In this study, we characterized two types of coated implant surfaces by scanning electron microscopy, energy dispersive spectrometry, and surface roughness testing. The effect of surface modification on early bone healing was then tested by using the rabbit tibia model to measure bone-to-implant contact ratios and removal torque values. These modified surfaces showed different characteristics in terms of surface topography, chemical composition, and surface roughness. However, no significant differences were found in the bone-to-implant contact and the resistance to removal torque between these surfaces. Both the coated implants may induce similar favorable early bone responses in terms of the early functioning and healing of dental implants even though they differed in their surface characteristics.

Results of bone regenerate study after osteosynthesis with bioinert and calcium phosphate-coated bioactive implants in experimental femoral neck fractures (experimental study

Directory of Open Access Journals (Sweden)

K. S. Kazanin

2015-01-01

Full Text Available Objective - to analyze the results of X-ray, cytomorphometric and immunohistochemistry experimental studies of bone regenerates after osteosynthesis with bioinert and calcium phosphate-coated bioactive implants. Material and methods. The study was conducted on experimental femoral neck fractures in rabbit males. Reparative osteogenesis processes were studied in groups of bioinert titanium implant osteosynthesis and calcium phosphate-coated bioactive titanium implant osteosynthesis. The animals were clinically followed-up during the postoperative period. X-ray, cytomorphometric and immunohistochemistry studies of samples extracted from femoral bones were conducted over time on days 1, 7, 14, 30 and 60. The animal experiments were kept and treated according to recommendations of international standards, Helsinki Declaration on animal welfare and approved by the local ethics committee. All surgeries were performed under anesthesia, and all efforts were made to minimize the suffering of the animals. Results. In the animal group without femoral neck fracture osteosynthesis, femoral neck pseudoarthrosis was observed at the end of the experiment. The results of cytomorphometric and immunohistochemistry studies conducted on day 60 of the experiment confirmed that the cellular composition of the bone regenerate in the group of calcium phosphate-coated bioactive titanium implants corresponded to a more mature bone tissue than in the group of bioinert titanium implants. Conclusion. The results of the statistical analysis of cytomorphometric and immunohistochemistry data show that the use of calcium phosphate-coated bioactive titanium implants allows to achieve significantly earlier bone tissue regeneration.

Porous SiO{sub 2} nanofiber grafted novel bioactive glass–ceramic coating: A structural scaffold for uniform apatite precipitation and oriented cell proliferation on inert implant

Energy Technology Data Exchange (ETDEWEB)

Das, Indranee [Nano-Structured Materials Division, CSIR-Central Glass and Ceramic Research Institute, Kolkata 700032 (India); De, Goutam, E-mail: gde@cgcri.res.in [Nano-Structured Materials Division, CSIR-Central Glass and Ceramic Research Institute, Kolkata 700032 (India); Hupa, Leena [Johan Gadolin Process Chemistry Centre, Åbo Akademi University, FI-20500 Åbo (Finland); Vallittu, Pekka K. [Turku Clinical Biomaterials Centre—TCBC, University of Turku, FI-20520 Turku (Finland); Institute of Dentistry, University of Turku, Department of Biomaterials Science and City of Turku, Welfare Division, Turku (Finland)

2016-05-01

A composite bioactive glass–ceramic coating grafted with porous silica nanofibers was fabricated on inert glass to provide a structural scaffold favoring uniform apatite precipitation and oriented cell proliferation. The coating surfaces were investigated thoroughly before and after immersion in simulated body fluid. In addition, the proliferation behavior of fibroblast cells on the surface was observed for several culture times. The nanofibrous exterior of this composite bioactive coating facilitated homogeneous growth of flake-like carbonated hydroxyapatite layer within a short period of immersion. Moreover, the embedded porous silica nanofibers enhanced hydrophilicity which is required for proper cell adhesion on the surface. The cells proliferated well following a particular orientation on the entire coating by the assistance of nanofibrous scaffold-like structural matrix. This newly engineered composite coating was effective in creating a biological structural matrix favorable for homogeneous precipitation of calcium phosphate, and organized cell growth on the inert glass surface. - Highlights: • Fabricated porous SiO{sub 2} nanofibers grafted composite bioactive glass–ceramic coating on inert glass. • The newly engineered coating facilitates uniformly dense apatite precipitation. • Embedded porous silica nanofibers enhance hydrophilicity of the coated surface. • Cells proliferate well on the entire coating following a particular orientation by the assistance of nanofibers. • The coatings have potential to be used as biological scaffold on the surface of implants.

Toughening and functionalization of bioactive ceramic and glass bone scaffolds by biopolymer coatings and infiltration: a review of the last 5 years.

Science.gov (United States)

Philippart, Anahí; Boccaccini, Aldo R; Fleck, Claudia; Schubert, Dirk W; Roether, Judith A

2015-01-01

Inorganic scaffolds with high interconnected porosity based on bioactive glasses and ceramics are prime candidates for applications in bone tissue engineering. These materials however exhibit relatively low fracture strength and high brittleness. A simple and effective approach to improve the toughness is to combine the basic scaffold structure with polymer coatings or through the formation of interpenetrating polymer-bioactive ceramic microstructures. The polymeric phase can additionally serve as a carrier for growth factors and therapeutic drugs, thus adding biological functionalities. The present paper reviews the state-of-the art in the field of polymer coated and infiltrated bioactive inorganic scaffolds. Based on the notable combination of bioactivity, improved mechanical properties and drug or growth factor delivery capability, this scaffold type is a candidate for bone and osteochondral regeneration strategies. Remaining challenges for the improvement of the materials are discussed and opportunities to broaden the application potential of this scaffold type are also highlighted.

Bioactivity response of Ta{sub 1-x}O{sub x} coatings deposited by reactive DC magnetron sputtering

Energy Technology Data Exchange (ETDEWEB)

Almeida Alves, C.F., E-mail: cristiana.alves@fisica.uminho.pt [GRF-CFUM, Physics Departament, University of Minho, Campus of Azurem, Guimaraes 4800-058 (Portugal); Cavaleiro, A. [SEG-CEMUC, Mechanical Engineering Department, University of Coimbra, Coimbra 3030-788 (Portugal); Carvalho, S. [GRF-CFUM, Physics Departament, University of Minho, Campus of Azurem, Guimaraes 4800-058 (Portugal); SEG-CEMUC, Mechanical Engineering Department, University of Coimbra, Coimbra 3030-788 (Portugal)

2016-01-01

The use of dental implants is sometimes accompanied by failure due to periimplantitis disease and subsequently poor esthetics when soft–hard tissue margin recedes. As a consequence, further research is needed for developing new bioactive surfaces able to enhance the osseous growth. Tantalum (Ta) is a promising material for dental implants since, comparing with titanium (Ti), it is bioactive and has an interesting chemistry which promotes the osseointegration. Another promising approach for implantology is the development of implants with oxidized surfaces since bone progenitor cells interact with the oxide layer forming a diffusion zone due to its ability to bind with calcium which promotes a stronger bond. In the present report Ta-based coatings were deposited by reactive DC magnetron sputtering onto Ti CP substrates in an Ar + O{sub 2} atmosphere. In order to assess the osteoconductive response of the studied materials, contact angle and in vitro tests of the samples immersed in Simulated Body Fluid (SBF) were performed. Structural results showed that oxide phases where achieved with larger amounts of oxygen (70 at.% O). More compact and smooth coatings were deposited by increasing the oxygen content. The as-deposited Ta coating presented the most hydrophobic character (100°); with increasing oxygen amount contact angles progressively diminished, down to the lowest measured value, 63°. The higher wettability is also accompanied by an increase on the surface energy. Bioactivity tests demonstrated that highest O-content coating, in good agreement with wettability and surface energy values, showed an increased affinity for apatite adhesion, with higher Ca/P ratio formation, when compared to the bare Ti substrates. - Highlights: • Ta{sub 1-x}O{sub x} coatings were deposited by reactive DC magnetron sputtering. • Amorphous oxide phases were achieved with higher oxygen amounts. • Contact angles progressively diminished, with increasing oxygen content. • Ta

In vivo bioactivity of titanium and fluorinated apatite coatings for orthopaedic implants: a vibrational study

Science.gov (United States)

Taddei, Paola; Tinti, Anna; Reggiani, Matteo; Monti, Patrizia; Fagnano, Concezio

2003-06-01

The bone integration of implants is a complex process which depends on chemical composition and surface morphology. To accelerate osteointegration, metal implants are coated with porous metal or apatites which have been reported to increase mineralisation, improving prosthesis fixation. To study the influence of composition and morphology on the in vivo bioactivity, titanium screws coated by Plasma Flame Spraying (PFS) with titanium or fluorinated apatite (K690) were implanted in sheep tibia and femur for 10 weeks and studied by micro-Raman and IR spectroscopy. The same techniques, together with thermogravimetry, were used for characterising the pre-coating K690 powder. Contrary to the manufacturer report, the K690 pre-coating revealed to be composed of a partially fluorinated apatite containing impurities of Ca(OH) 2 and CaCO 3. By effect of PFS, the impurities were decomposed and the crystallinity degree of the coating was found to decrease. The vibrational spectra recorded on the implanted screws revealed the presence of newly formed bone; for the K690-coated screws at least, a high level of osteointegration was evidenced.

Amino acid sequences mediating vascular cell adhesion molecule 1 binding to integrin alpha 4: homologous DSP sequence found for JC polyoma VP1 coat protein

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Michael Andrew Meyer

2013-07-01

Full Text Available The JC polyoma viral coat protein VP1 was analyzed for amino acid sequences homologies to the IDSP sequence which mediates binding of VLA-4 (integrin alpha 4 to vascular cell adhesion molecule 1. Although the full sequence was not found, a DSP sequence was located near the critical arginine residue linked to infectivity of the virus and binding to sialic acid containing molecules such as integrins (3. For the JC polyoma virus, a DSP sequence was found at residues 70, 71 and 72 with homology also noted for the mouse polyoma virus and SV40 virus. Three dimensional modeling of the VP1 molecule suggests that the DSP loop has an accessible site for interaction from the external side of the assembled viral capsid pentamer.

Investigation of morphology and bioactive properties of composite coating of HA/vinyl acetate on pure titanium

International Nuclear Information System (INIS)

Afshar, Abdollahe; Yousefpour, Mardali; Xiudong, Yang; Li Xudong; Yang Bangcheng; Wu Yao; Chen Jiyong; Zhang Xingdong

2006-01-01

Electrochemical co-deposition approach was expanded to prepare composite bio-ceramic coating of hydroxyapatite (HA)/polyvinyl acetate on the surface of titanium. The role is to improve the bioactive and crystallization properties. The results of XRD, XPS, SEM and TEM characterization showed that by increasing amount of vinyl acetate in the composite bio-ceramic coating before and after immersing in the simulated body fluid (SBF), an oriented growth of HA planes on the (0 0 2) direction had been observed on titanium substrate. Also significant surface morphology changes were obtained

A bioactive coating with submicron-sized titania crystallites fabricated by induction heating of titanium after tensile deformations.

Science.gov (United States)

Li, Ning-Bo; Xu, Wen-Hua; Xiao, Gui-Yong; Zhao, Jun-Han; Lu, Yu-Peng

2017-11-01

Thermal oxidation technology was widely investigated as one of effective surface modification method for improving the bioactivity and biocompatibility of titanium and its alloys. In this work, the induction heat oxidization method, a fast, efficient, economical and environmental protective technology, was applied to prepare the submicron-morphological oxide coating with variable rutile TiO 2 equiaxed crystallites on the surface of pure Ti substrates after cold-drawing with 10-20% deformations. The results showed the plastic-deformed Ti cylinders recrystallized during induction heating treatment (IHT) for 10-20s which resulted in evolution of microstructures as well as slight improvement of microhardness. The surface characteristics of TiO 2 crystallites in oxidation layers were determined by the microstructural evolutions of Ti substrate in terms of the nucleation and growth of TiO 2 crystallites. Specially, the oxidized surface with 50-75nm roughness and more uniform and finer equiaxed oxide grains remarkablely improved the apatite deposition after bioactive evaluation in 1.5 × SBF for 7 days. This work provided a potential method to create controlled bioactive oxide coatings with submicro-/nano-scaled TiO 2 crystallites on titanium substrate in terms of the role of metallographic microstructure in the formation process of titanium oxides. Copyright © 2017 Elsevier Ltd. All rights reserved.

Silk coating on a bioactive ceramic scaffold for bone regeneration: effective enhancement of mechanical and in vitro osteogenic properties towards load-bearing applications.

Science.gov (United States)

Li, Jiao Jiao; Roohani-Esfahani, Seyed-Iman; Kim, Kyungsook; Kaplan, David L; Zreiqat, Hala

2017-06-01

Bioactive ceramic scaffolds represent competitive choices for clinical bone reconstruction, but their widespread use is restricted by inherent brittleness and weak mechanical performance under load. This study reports the development of strong and tough bioactive scaffolds suitable for use in load-bearing bone reconstruction. A strong and bioactive ceramic scaffold (strontium-hardystonite-gahnite) is combined with single and multiple coating layers of silk fibroin to enhance its toughness, producing composite scaffolds which match the mechanical properties of cancellous bone and show enhanced capacity to promote in vitro osteogenesis. Also reported for the first time is a comparison of the coating effects obtained when a polymeric material is coated on ceramic scaffolds with differing microstructures, namely the strontium-hardystonite-gahnite scaffold with high-density struts as opposed to a conventional ceramic scaffold, such as biphasic calcium phosphate, with low-density struts. The results show that silk coating on a unique ceramic scaffold can lead to simple and effective enhancement of its mechanical and biological properties to suit a wider range of applications in clinical bone reconstruction, and also establish the influence of ceramic microstructure on the effectiveness of silk coating as a method of reinforcement when applied to different types of ceramic bone graft substitutes. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Bioactive glass-chitosan composite coatings on PEEK: Effects of surface wettability and roughness on the interfacial fracture resistance and in vitro cell response

Science.gov (United States)

Hong, Wei; Guo, Fangwei; Chen, Jianwei; Wang, Xin; Zhao, Xiaofeng; Xiao, Ping

2018-05-01

To improve the osteointegration of polyetheretherketone (PEEK) spinal fusions, the 45S5 bioactive glass® (BG)-chitosan (CH) composite was used to coat the PEEK by a dip-coating method at room temperature. A robust bonding between the BG-CH composite coating and the PEEK was achieved by a combined surface treatment of sand blasting and acid etching. The effects of surface wettability and surface roughness on the adhesion of the BG-CH composite coating were characterized by fracture resistance (Gc), respectively, measured by four-point bending tests. Compared with the surface polar energy (wettability), the surface roughness (>3 μm) played a more important role for the increase in Gc values by means of crack shielding effect under the mixed mode stress. The maximum adhesion strength (σ) of the coatings on the modified PEEK measured by the tensile pull-off test was about 5.73 MPa. The in vitro biocompatibilities of PEEK, including cell adhesion, cell proliferation, differentiation, and bioactivity in the stimulated body fluid (SBF), were enhanced by the presence of BG-CH composite coatings, which also suggested that this composite coating method could provide an effective solution for the weak PEEK-bone integration.

Calcium phosphate/chitosan composite coating: Effect of different concentrations of Mg2+ in the m-SBF on its bioactivity

International Nuclear Information System (INIS)

Zhang, Jie; Dai, Changsong; Wei, Jie; Wen, Zhaohui; Zhang, Shujuan; Lin, Lemin

2013-01-01

The purpose of this study was to investigate the effect of different concentration of Mg 2+ in a modified simulated body fluid (m-SBF) on the bioactivity of calcium phosphate/chitosan composite coating. Calcium phosphate/chitosan composite coating was prepared on graphite substrate via electrophoretic deposition (EPD) followed by conversion in a phosphate buffer solution (PBS). The obtained samples were soaked in the m-SBF containing different concentration of Mg 2+ for different times. And then, the composite coatings were assessed using X-ray diffractometer (XRD), Fourier-transformed infrared spectroscopy (FTIR), Raman spectra, and scanning electron microscope (SEM) with an energy dispersive spectrometer (EDS). The soaking solution was evaluated by inductively coupled plasma optical emission spectrometer (ICP-OES) test. The analytical results showed that hydroxyapatite (HA) and bone-like apatite (HCA) grew on the surface of calcium phosphate/chitosan composite coating after incubation in different m-SBF. With Mg 2+ concentration in m-SBF increased from 1× Mg to 10× Mg, HA in the composite coating first presented a dissolving process and then a precipitating one slowly, while HCA presented a growing trend, continuously. The increasing of Mg 2+ concentration in the m-SBF inhibited the total growing process of HA and HCA as a whole. The structure of the composite coating changed from spherical into irregular morphology with the concentration of Mg 2+ increasing from 1× Mg to 10× Mg. Over all, with the Mg 2+ concentration increasing, the bioactivity of calcium phosphate/chitosan composite coating tended to decrease.

Calcium phosphate/chitosan composite coating: Effect of different concentrations of Mg2+ in the m-SBF on its bioactivity

Science.gov (United States)

Zhang, Jie; Dai, Changsong; Wei, Jie; Wen, Zhaohui; Zhang, Shujuan; Lin, Lemin

2013-09-01

The purpose of this study was to investigate the effect of different concentration of Mg2+ in a modified simulated body fluid (m-SBF) on the bioactivity of calcium phosphate/chitosan composite coating. Calcium phosphate/chitosan composite coating was prepared on graphite substrate via electrophoretic deposition (EPD) followed by conversion in a phosphate buffer solution (PBS). The obtained samples were soaked in the m-SBF containing different concentration of Mg2+ for different times. And then, the composite coatings were assessed using X-ray diffractometer (XRD), Fourier-transformed infrared spectroscopy (FTIR), Raman spectra, and scanning electron microscope (SEM) with an energy dispersive spectrometer (EDS). The soaking solution was evaluated by inductively coupled plasma optical emission spectrometer (ICP-OES) test. The analytical results showed that hydroxyapatite (HA) and bone-like apatite (HCA) grew on the surface of calcium phosphate/chitosan composite coating after incubation in different m-SBF. With Mg2+ concentration in m-SBF increased from 1× Mg to 10× Mg, HA in the composite coating first presented a dissolving process and then a precipitating one slowly, while HCA presented a growing trend, continuously. The increasing of Mg2+ concentration in the m-SBF inhibited the total growing process of HA and HCA as a whole. The structure of the composite coating changed from spherical into irregular morphology with the concentration of Mg2+ increasing from 1× Mg to 10× Mg. Over all, with the Mg2+ concentration increasing, the bioactivity of calcium phosphate/chitosan composite coating tended to decrease.

Comparison study of biomimetic strontium-doped calcium phosphate coatings by electrochemical deposition and air plasma spray: morphology, composition and bioactive performance.

Science.gov (United States)

Li, Ling; Lu, Xia; Meng, Yizhi; Weyant, Christopher M

2012-10-01

In this study, strontium-doped calcium phosphate coatings were deposited by electrochemical deposition and plasma spray under different process parameters to achieve various coating morphologies. The coating composition was investigated by energy dispersive X-ray spectroscopy and X-ray diffraction. The surface morphologies of the coatings were studied through scanning electron microscopy while the cytocompatibility and bioactivity of the strontium-doped calcium phosphate coatings were evaluated using bone cell culture using MC3T3-E1 osteoblast-like cells. The addition of strontium leads to enhanced proliferation suggesting the possible benefits of strontium incorporation in calcium phosphate coatings. The morphology and composition of deposited coatings showed a strong influence on the growth of cells.

Improved surface bioactivity of stainless steel substrates using osteocalcin mimetic peptide

International Nuclear Information System (INIS)

Hosseini, Samaneh; Naderi-Manesh, Hossein; Vali, Hojatollah; Faghihi, Shahab

2014-01-01

Although stainless steel has a good biocompatibility for most clinical cases, the higher tissue response (bone bonding property) is required in orthopedic field. In this study, to improve bone-bonding ability of stainless steel substrates, a specific sequence of osteocalcin mimetic peptide is used as bioactive coating material to biochemically modify the surface of metallic samples. This sequence consists of thirteen amino acids present in the first helix of osteocalcin is synthesized in amidic form and physically adsorbed on the surface of 316LS (316 low carbon surgical grade) stainless steel substrates. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) are used to characterize the surface of peptide coated and uncoated substrates. The bioactivity and bone bonding ability of coated and uncoated substrates are assessed by level of hydroxyapatite formation, using transmission electron microscopy (TEM), energy-dispersive x-ray (EDS), and scanning electron microscopy (SEM). The pre-osteoblast cell attachment and proliferation are also evaluated by MTT assay. The results show that the surface of coated sample is homogenously covered by the peptide and display a rougher surface relative to uncoated sample. TEM images reveal the formation of plate-like hydroxyapatite crystals in the presence of the peptide and an amorphous calcium phosphate phase without the peptide. Pre-osteoblast cells proliferation is significantly higher on the surface of peptide coated substrate, while cell attachment remains unaffected by the peptide coatings. Pre-osteoblast cells also demonstrate a higher degree of spreading on the surface of coated sample. It is believed that osteocalcin mimetic peptide improve surface bioactivity and promote hydroxyapatite crystal formation may lead to increased mineralization and bone formation on the surface of metallic biomedical devices. - Graphical abstract: A peptide sequence located in the first helix of OC is selected based on its

Improved surface bioactivity of stainless steel substrates using osteocalcin mimetic peptide

Energy Technology Data Exchange (ETDEWEB)

Hosseini, Samaneh [Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Tissue Engineering and Biomaterials Division, National Institute of Genetic Engineering and Biotechnology, Tehran 14965/161 (Iran, Islamic Republic of); Naderi-Manesh, Hossein, E-mail: naderman@modares.ac.ir [Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Vali, Hojatollah [Department of Anatomy and Cell Biology, McGill University, 3640 University Street, Montréal, QC H3A 0C7 (Canada); Faghihi, Shahab, E-mail: sfaghihi@nigeb.ac.ir [Tissue Engineering and Biomaterials Division, National Institute of Genetic Engineering and Biotechnology, Tehran 14965/161 (Iran, Islamic Republic of)

2014-02-14

Although stainless steel has a good biocompatibility for most clinical cases, the higher tissue response (bone bonding property) is required in orthopedic field. In this study, to improve bone-bonding ability of stainless steel substrates, a specific sequence of osteocalcin mimetic peptide is used as bioactive coating material to biochemically modify the surface of metallic samples. This sequence consists of thirteen amino acids present in the first helix of osteocalcin is synthesized in amidic form and physically adsorbed on the surface of 316LS (316 low carbon surgical grade) stainless steel substrates. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) are used to characterize the surface of peptide coated and uncoated substrates. The bioactivity and bone bonding ability of coated and uncoated substrates are assessed by level of hydroxyapatite formation, using transmission electron microscopy (TEM), energy-dispersive x-ray (EDS), and scanning electron microscopy (SEM). The pre-osteoblast cell attachment and proliferation are also evaluated by MTT assay. The results show that the surface of coated sample is homogenously covered by the peptide and display a rougher surface relative to uncoated sample. TEM images reveal the formation of plate-like hydroxyapatite crystals in the presence of the peptide and an amorphous calcium phosphate phase without the peptide. Pre-osteoblast cells proliferation is significantly higher on the surface of peptide coated substrate, while cell attachment remains unaffected by the peptide coatings. Pre-osteoblast cells also demonstrate a higher degree of spreading on the surface of coated sample. It is believed that osteocalcin mimetic peptide improve surface bioactivity and promote hydroxyapatite crystal formation may lead to increased mineralization and bone formation on the surface of metallic biomedical devices. - Graphical abstract: A peptide sequence located in the first helix of OC is selected based on its

Effect of layer-by-layer coatings and localization of antioxidant on oxidative stability of a model encapsulated bioactive compound in oil-in-water emulsions.

Science.gov (United States)

Pan, Yuanjie; Nitin, N

2015-11-01

Oxidation of encapsulated bioactives in emulsions is one of the key challenges that limit shelf-life of many emulsion containing products. This study seeks to quantify the role of layer-by-layer coatings and localization of antioxidant molecules at the emulsion interface in influencing oxidation of the encapsulated bioactives. Oxidative barrier properties of the emulsions were simulated by measuring the rate of reaction of peroxyl radicals generated in the aqueous phase with the encapsulated radical sensitive dye in the lipid core of the emulsions. The results of peroxyl radical permeation were compared to the stability of encapsulated retinol (model bioactive) in emulsions. To evaluate the role of layer-by-layer coatings in influencing oxidative barrier properties, radical permeation rates and retinol stability were evaluated in emulsion formulations of SDS emulsion and SDS emulsion with one or two layers of polymers (ϵ-polylysine and dextran sulfate) coated at the interface. To localize antioxidant molecules to the interface, gallic acid (GA) was chemically conjugated with ϵ-polylysine and subsequently deposited on SDS emulsion based on electrostatic interactions. Emulsion formulations with localized GA molecules at the interface were compared with SDS emulsion with GA molecules in the bulk aqueous phase. The results of this study demonstrate the advantage of localization of antioxidant at the interface and the limited impact of short chain polymer coatings at the interface of emulsions in reducing permeation of radicals and oxidation of a model encapsulated bioactive in oil-in-water emulsions. Copyright © 2015 Elsevier B.V. All rights reserved.

Corrosion and bioactivity performance of graphene oxide coating on TiNb shape memory alloys in simulated body fluid.

Science.gov (United States)

Saud, Safaa N; Hosseinian S, Raheleh; Bakhsheshi-Rad, H R; Yaghoubidoust, F; Iqbal, N; Hamzah, E; Ooi, C H Raymond

2016-11-01

In the present work, the microstructure, corrosion, and bioactivity of graphene oxide (GO) coating on the laser-modified and -unmodified surfaces of TiNb shape memory alloys (SMAs) were investigated. The surface morphology and chemical composition was examined using field emission scanning electron microscopy (FE-SEM) and X-ray diffraction (XRD). The surface modification was carried out via a femtosecond laser with the aim to increase the surface roughness, and thus increase the adhesion property. FE-SEM analysis of the laser-treated Ti-30at.% Nb revealed the increase in surface roughness and oxygen/nitrogen containing groups on the Ti-30at.% Nb surface after being surface modified via a femtosecond laser. Furthermore, the thickness of GO was increased from 35μm to 45μm after the surface was modified. Potentiodynamic polarisation and electrochemical impedance spectroscopy studies revealed that both the GO and laser/GO-coated samples exhibited higher corrosion resistance than that of the uncoated TiNb SMA sample. However, the laser/GO-coated sample presented the highest corrosion resistance in SBF at 37°C. In addition, during soaking in the simulated body fluid (SBF), both the GO and laser/GO coating improved the formation of apatite layer. Based on the bioactivity results, the GO coating exhibited a remarkable antibacterial activity against gram-negative bacteria compared with the uncoated. In conclusion, the present results indicate that Ti-30at.% Nb SMAs may be promising alternatives to NiTi for certain biomedical applications. Copyright © 2016 Elsevier B.V. All rights reserved.

Sol-gel derived bioactive coating on zirconia: Effect on flexural strength and cell proliferation.

Science.gov (United States)

Shahramian, Khalil; Leminen, Heidi; Meretoja, Ville; Linderbäck, Paula; Kangasniemi, Ilkka; Lassila, Lippo; Abdulmajeed, Aous; Närhi, Timo

2017-11-01

The purpose of this study was to evaluate the effect of sol-gel derived bioactive coatings on the biaxial flexural strength and fibroblast proliferation of zirconia, aimed to be used as an implant abutment material. Yttrium stabilized zirconia disc-shaped specimens were cut, ground, sintered, and finally cleansed ultrasonically in each of acetone and ethanol for 5 minutes. Three experimental groups (n = 15) were fabricated, zirconia with sol-gel derived titania (TiO 2 ) coating, zirconia with sol-gel derived zirconia (ZrO 2 ) coating, and non-coated zirconia as a control. The surfaces of the specimens were analyzed through images taken using a scanning electron microscope (SEM), and a non-contact tapping mode atomic force microscope (AFM) was used to record the surface topography and roughness of the coated specimens. Biaxial flexural strength values were determined using the piston-on-three ball technique. Human gingival fibroblast proliferation on the surface of the specimens was evaluated using AlamarBlue assay™. Data were analyzed using a one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test. Additionally, the biaxial flexural strength data was also statistically analyzed with the Weibull distribution. The biaxial flexural strength of zirconia specimens was unaffected (p > 0.05). Weibull modulus of TiO 2 coated and ZrO 2 coated groups (5.7 and 5.4, respectively) were lower than the control (8.0). Specimens coated with ZrO 2 showed significantly lower fibroblast proliferation compared to other groups (p sol-gel derived coatings have no influence on the flexural strength of zirconia. ZrO 2 coated specimens showed significantly lower cell proliferation after 12 days than TiO 2 coated or non-coated control. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2401-2407, 2017. © 2016 Wiley Periodicals, Inc.

Calcium phosphate/chitosan composite coating: Effect of different concentrations of Mg{sup 2+} in the m-SBF on its bioactivity

Energy Technology Data Exchange (ETDEWEB)

Zhang, Jie [School of Chemistry Engineering and Technology, Harbin Institute of Technology, Harbin 150001 (China); Pharmacy College, Jiamusi University, Jiamusi 154007 (China); Dai, Changsong, E-mail: changsd@hit.edu.cn [School of Chemistry Engineering and Technology, Harbin Institute of Technology, Harbin 150001 (China); Wei, Jie [School of Chemistry Engineering and Technology, Harbin Institute of Technology, Harbin 150001 (China); School of Chemistry and Bioengineering, Suzhou Science Technology University, Suzhou 215009 (China); Wen, Zhaohui, E-mail: wenzhaohui1968@163.com [Department of neuro intern, First Affiliated Hospital of Harbin Medical University, Harbin 150001 (China); Zhang, Shujuan; Lin, Lemin [Department of neuro intern, First Affiliated Hospital of Harbin Medical University, Harbin 150001 (China)

2013-09-01

The purpose of this study was to investigate the effect of different concentration of Mg{sup 2+} in a modified simulated body fluid (m-SBF) on the bioactivity of calcium phosphate/chitosan composite coating. Calcium phosphate/chitosan composite coating was prepared on graphite substrate via electrophoretic deposition (EPD) followed by conversion in a phosphate buffer solution (PBS). The obtained samples were soaked in the m-SBF containing different concentration of Mg{sup 2+} for different times. And then, the composite coatings were assessed using X-ray diffractometer (XRD), Fourier-transformed infrared spectroscopy (FTIR), Raman spectra, and scanning electron microscope (SEM) with an energy dispersive spectrometer (EDS). The soaking solution was evaluated by inductively coupled plasma optical emission spectrometer (ICP-OES) test. The analytical results showed that hydroxyapatite (HA) and bone-like apatite (HCA) grew on the surface of calcium phosphate/chitosan composite coating after incubation in different m-SBF. With Mg{sup 2+} concentration in m-SBF increased from 1× Mg to 10× Mg, HA in the composite coating first presented a dissolving process and then a precipitating one slowly, while HCA presented a growing trend, continuously. The increasing of Mg{sup 2+} concentration in the m-SBF inhibited the total growing process of HA and HCA as a whole. The structure of the composite coating changed from spherical into irregular morphology with the concentration of Mg{sup 2+} increasing from 1× Mg to 10× Mg. Over all, with the Mg{sup 2+} concentration increasing, the bioactivity of calcium phosphate/chitosan composite coating tended to decrease.

Porous SiO2 nanofiber grafted novel bioactive glass-ceramic coating: A structural scaffold for uniform apatite precipitation and oriented cell proliferation on inert implant.

Science.gov (United States)

Das, Indranee; De, Goutam; Hupa, Leena; Vallittu, Pekka K

2016-05-01

A composite bioactive glass-ceramic coating grafted with porous silica nanofibers was fabricated on inert glass to provide a structural scaffold favoring uniform apatite precipitation and oriented cell proliferation. The coating surfaces were investigated thoroughly before and after immersion in simulated body fluid. In addition, the proliferation behavior of fibroblast cells on the surface was observed for several culture times. The nanofibrous exterior of this composite bioactive coating facilitated homogeneous growth of flake-like carbonated hydroxyapatite layer within a short period of immersion. Moreover, the embedded porous silica nanofibers enhanced hydrophilicity which is required for proper cell adhesion on the surface. The cells proliferated well following a particular orientation on the entire coating by the assistance of nanofibrous scaffold-like structural matrix. This newly engineered composite coating was effective in creating a biological structural matrix favorable for homogeneous precipitation of calcium phosphate, and organized cell growth on the inert glass surface. Copyright © 2016 Elsevier B.V. All rights reserved.

Bioactive Glass Nanopowder for theTreatment of Oral Bone Defects

Directory of Open Access Journals (Sweden)

MH. Fathi

2007-09-01

Full Text Available Objective: Osseous defects around dental implants are often seen when implants are placed in areas with inadequate alveolar bone, or around failing implants. Bone regenera-tion in these areas using bone grafts or its substitutes may improve dental implants prog-nosis. The aim of this study was to prepare and characterize the bioactive glass nanopow-der and development of its coating for treatment of oral bone defects.Materials and Methods: Bioactive bioglass coating was made on stainless steel plates by sol-gel technique. The powder shape and size was evaluated by transmission electron mi-cropscopy, and thermal properties studied using differential thermal analysis (DTA. Structural characterization techniques (XRD were used to analyze and study the structure and phase present in the prepared bioactive glass nanopowder. This nanopowder was immersed in the simulated body fluid (SBF solution. Fourier transform infrared spec-troscopy (FTIR was utilized to recognize and confirm the formation of apatite layer on prepared bioactive glass nanopowder.Results: The bioglass powder size was less than 100 nanometers which was necessary for better bioactivity, and preparing a homogeneous coating. The formation of apatite layer confirmed the bioactivity of the bioglass nanopowder. Crack-free and homogeneous bioglass coatings were achieved with no observable defects.Conclusion: It was concluded that the prepared bioactive glass nanopowder could be more effective as a bone replacement material than conventional bioactive glass to pro-mote bone formation in osseous defects. The prepared bioactive glass nanopowder could be more useful for treatment of oral bone defects compare to conventional hydroxyapatite or bioactive glass.

Bioactive lipid coating of bone allografts directs engraftment and fate determination of bone marrow-derived cells in rat GFP chimeras

OpenAIRE

Das, Anusuya; Segar, Claire E.; Chu, Yihsuan; Wang, Tiffany W.; Lin, Yong; Yang, Chunxi; Du, Xeujun; Ogle, Roy C.; Cui, Quanjun; Botchwey, Edward A.

2015-01-01

Bone grafting procedures are performed to treat wounds incurred during wartime trauma, accidents, and tumor resections. Endogenous mechanisms of repair are often insufficient to ensure integration between host and donor bone and subsequent restoration of function. We investigated the role that bone marrow-derived cells play in bone regeneration and sought to increase their contributions by functionalizing bone allografts with bioactive lipid coatings. Polymer-coated allografts were used to lo...

Integrin-directed modulation of macrophage responses to biomaterials.

Science.gov (United States)

Zaveri, Toral D; Lewis, Jamal S; Dolgova, Natalia V; Clare-Salzler, Michael J; Keselowsky, Benjamin G

2014-04-01

Macrophages are the primary mediator of chronic inflammatory responses to implanted biomaterials, in cases when the material is either in particulate or bulk form. Chronic inflammation limits the performance and functional life of numerous implanted medical devices, and modulating macrophage interactions with biomaterials to mitigate this response would be beneficial. The integrin family of cell surface receptors mediates cell adhesion through binding to adhesive proteins nonspecifically adsorbed onto biomaterial surfaces. In this work, the roles of integrin Mac-1 (αMβ2) and RGD-binding integrins were investigated using model systems for both particulate and bulk biomaterials. Specifically, the macrophage functions of phagocytosis and inflammatory cytokine secretion in response to a model particulate material, polystyrene microparticles were investigated. Opsonizing proteins modulated microparticle uptake, and integrin Mac-1 and RGD-binding integrins were found to control microparticle uptake in an opsonin-dependent manner. The presence of adsorbed endotoxin did not affect microparticle uptake levels, but was required for the production of inflammatory cytokines in response to microparticles. Furthermore, it was demonstrated that integrin Mac-1 and RGD-binding integrins influence the in vivo foreign body response to a bulk biomaterial, subcutaneously implanted polyethylene terephthalate. A thinner foreign body capsule was formed when integrin Mac-1 was absent (~30% thinner) or when RGD-binding integrins were blocked by controlled release of a blocking peptide (~45% thinner). These findings indicate integrin Mac-1 and RGD-binding integrins are involved and may serve as therapeutic targets to mitigate macrophage inflammatory responses to both particulate and bulk biomaterials. Copyright © 2014 Elsevier Ltd. All rights reserved.

Biocatalytic coatings for air pollution control: a proof of concept study on VOC biodegradation.

Science.gov (United States)

Estrada, José M; Bernal, Oscar I; Flickinger, Michael C; Muñoz, Raúl; Deshusses, Marc A

2015-02-01

Although biofilm-based biotechnologies exhibit a large potential as solutions for off-gas treatment, the high water content of biofilms often causes pollutant mass transfer limitations, which ultimately limit their widespread application. The present study reports on the proof of concept of the applicability of bioactive latex coatings for air pollution control. Toluene vapors served as a model volatile organic compound (VOC). The results showed that Pseudomonas putida F1 cells could be successfully entrapped in nanoporous latex coatings while preserving their toluene degradation activity. Bioactive latex coatings exhibited toluene specific biodegradation rates 10 times higher than agarose-based biofilms, because the thin coatings were less subject to diffusional mass transfer limitations. Drying and pollutant starvation were identified as key factors inducing a gradual deterioration of the biodegradation capacity in these innovative coatings. This study constitutes the first application of bioactive latex coatings for VOC abatement. These coatings could become promising means for air pollution control. © 2014 Wiley Periodicals, Inc.

Bioactive glass-ceramic coating for enhancing the in vitro corrosion resistance of biodegradable Mg alloy

Science.gov (United States)

Ye, Xinyu; Cai, Shu; Dou, Ying; Xu, Guohua; Huang, Kai; Ren, Mengguo; Wang, Xuexin

2012-10-01

In this work, a bioactive 45S5 glass-ceramic coating was synthesized on magnesium (Mg) alloy substrate by using a sol-gel dip-coating method, to improve the initial corrosion resistance of AZ31 Mg alloy. The surface morphology and phase composition of the glass-ceramic coating were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The coating composed of amorphous phase and crystalline phase Na2Ca2Si3O9, with the thickness of ∼1.0 μm, exhibited a uniform and crack-free surface morphology. The corrosion behavior of the uncoated and coated Mg alloy substrates was investigated by the electrochemical measurements and immersion tests in simulated body fluid (SBF). Potentiodynamic polarization tests recorded an increase of potential (Ecorr) form -1.60 V to -1.48 V, and a reduction of corrosion current density (icorr) from 4.48 μA cm-2 to 0.16 μA cm-2, due to the protection provided by the glass-ceramic coating. Immersion tests also showed the markedly improved corrosion resistance of the coated sample over the immersion period of 7 days. Moreover, after 14 days of immersion in SBF, the corrosion resistance of the coated sample declined due to the cracking of the glass-ceramic coating, which was confirmed by electrochemical impedance spectroscopy (EIS) analysis. The results suggested that the 45S5 glass-ceramic coated Mg alloy could provide a suitable corrosion behavior for use as degradable implants.

Mesoporous Bioactive Glass Functionalized 3D Ti-6Al-4V Scaffolds with Improved Surface Bioactivity.

Science.gov (United States)

Ye, Xiaotong; Leeflang, Sander; Wu, Chengtie; Chang, Jiang; Zhou, Jie; Huan, Zhiguang

2017-10-27

Porous Ti-6Al-4V scaffolds fabricated by means of selective laser melting (SLM), having controllable geometrical features and preferable mechanical properties, have been developed as a class of biomaterials that hold promising potential for bone repair. However, the inherent bio-inertness of the Ti-6Al-4V alloy as the matrix of the scaffolds results in a lack in the ability to stimulate bone ingrowth and regeneration. The aim of the present study was to develop a bioactive coating on the struts of SLM Ti-6Al-4V scaffolds in order to add the desired surface osteogenesis ability. Mesoporous bioactive glasses (MBGs) coating was applied on the strut surfaces of the SLM Ti-6Al-4V scaffolds through spin coating, followed by a heat treatment. It was found that the coating could maintain the characteristic mesoporous structure and chemical composition of MBG, and establish good interfacial adhesion to the Ti-6Al-4V substrate. The compressive strength and pore interconnectivity of the scaffolds were not affected by the coating. Moreover, the results obtained from in vitro cell culture experiments demonstrated that the attachment, proliferation, and differentiation of human bone marrow stromal cells (hBMSCs) on the MBG-coated Ti-6Al-4V scaffolds were improved as compared with those on the conventional bioactive glass (BG)-coated Ti-6Al-4V scaffolds and bare-metal Ti-6Al-4V scaffolds. Our results demonstrated that the MBG coating by using the spinning coating method could be an effective approach to achieving enhanced surface biofunctionalization for SLM Ti-6Al-4V scaffolds.

Mesoporous Bioactive Glass Functionalized 3D Ti-6Al-4V Scaffolds with Improved Surface Bioactivity

Directory of Open Access Journals (Sweden)

Xiaotong Ye

2017-10-01

Full Text Available Porous Ti-6Al-4V scaffolds fabricated by means of selective laser melting (SLM, having controllable geometrical features and preferable mechanical properties, have been developed as a class of biomaterials that hold promising potential for bone repair. However, the inherent bio-inertness of the Ti-6Al-4V alloy as the matrix of the scaffolds results in a lack in the ability to stimulate bone ingrowth and regeneration. The aim of the present study was to develop a bioactive coating on the struts of SLM Ti-6Al-4V scaffolds in order to add the desired surface osteogenesis ability. Mesoporous bioactive glasses (MBGs coating was applied on the strut surfaces of the SLM Ti-6Al-4V scaffolds through spin coating, followed by a heat treatment. It was found that the coating could maintain the characteristic mesoporous structure and chemical composition of MBG, and establish good interfacial adhesion to the Ti-6Al-4V substrate. The compressive strength and pore interconnectivity of the scaffolds were not affected by the coating. Moreover, the results obtained from in vitro cell culture experiments demonstrated that the attachment, proliferation, and differentiation of human bone marrow stromal cells (hBMSCs on the MBG-coated Ti-6Al-4V scaffolds were improved as compared with those on the conventional bioactive glass (BG-coated Ti-6Al-4V scaffolds and bare-metal Ti-6Al-4V scaffolds. Our results demonstrated that the MBG coating by using the spinning coating method could be an effective approach to achieving enhanced surface biofunctionalization for SLM Ti-6Al-4V scaffolds.

Bioactive glass–ceramic coating for enhancing the in vitro corrosion resistance of biodegradable Mg alloy

International Nuclear Information System (INIS)

Ye Xinyu; Cai Shu; Dou Ying; Xu Guohua; Huang Kai; Ren Mengguo; Wang Xuexin

2012-01-01

Highlights: ► Sol–gel derived 45S5 glass–ceramic coating was prepared on Mg alloy substrate. ► The corrosion resistance of glass–ceramic coated Mg alloy was markedly improved. ► The corrosion behavior of the coated sample varied due to the cracking of coating. - Abstract: In this work, a bioactive 45S5 glass–ceramic coating was synthesized on magnesium (Mg) alloy substrate by using a sol–gel dip-coating method, to improve the initial corrosion resistance of AZ31 Mg alloy. The surface morphology and phase composition of the glass–ceramic coating were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The coating composed of amorphous phase and crystalline phase Na 2 Ca 2 Si 3 O 9 , with the thickness of ∼1.0 μm, exhibited a uniform and crack-free surface morphology. The corrosion behavior of the uncoated and coated Mg alloy substrates was investigated by the electrochemical measurements and immersion tests in simulated body fluid (SBF). Potentiodynamic polarization tests recorded an increase of potential (E corr ) form −1.60 V to −1.48 V, and a reduction of corrosion current density (i corr ) from 4.48 μA cm −2 to 0.16 μA cm −2 , due to the protection provided by the glass–ceramic coating. Immersion tests also showed the markedly improved corrosion resistance of the coated sample over the immersion period of 7 days. Moreover, after 14 days of immersion in SBF, the corrosion resistance of the coated sample declined due to the cracking of the glass–ceramic coating, which was confirmed by electrochemical impedance spectroscopy (EIS) analysis. The results suggested that the 45S5 glass–ceramic coated Mg alloy could provide a suitable corrosion behavior for use as degradable implants.

Conservation of the human integrin-type beta-propeller domain in bacteria.

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Bhanupratap Chouhan

Full Text Available Integrins are heterodimeric cell-surface receptors with key functions in cell-cell and cell-matrix adhesion. Integrin α and β subunits are present throughout the metazoans, but it is unclear whether the subunits predate the origin of multicellular organisms. Several component domains have been detected in bacteria, one of which, a specific 7-bladed β-propeller domain, is a unique feature of the integrin α subunits. Here, we describe a structure-derived motif, which incorporates key features of each blade from the X-ray structures of human αIIbβ3 and αVβ3, includes elements of the FG-GAP/Cage and Ca(2+-binding motifs, and is specific only for the metazoan integrin domains. Separately, we searched for the metazoan integrin type β-propeller domains among all available sequences from bacteria and unicellular eukaryotic organisms, which must incorporate seven repeats, corresponding to the seven blades of the β-propeller domain, and so that the newly found structure-derived motif would exist in every repeat. As the result, among 47 available genomes of unicellular eukaryotes we could not find a single instance of seven repeats with the motif. Several sequences contained three repeats, a predicted transmembrane segment, and a short cytoplasmic motif associated with some integrins, but otherwise differ from the metazoan integrin α subunits. Among the available bacterial sequences, we found five examples containing seven sequential metazoan integrin-specific motifs within the seven repeats. The motifs differ in having one Ca(2+-binding site per repeat, whereas metazoan integrins have three or four sites. The bacterial sequences are more conserved in terms of motif conservation and loop length, suggesting that the structure is more regular and compact than those example structures from human integrins. Although the bacterial examples are not full-length integrins, the full-length metazoan-type 7-bladed β-propeller domains are present, and

Disintegrins: integrin selective ligands which activate integrin-coupled signaling and modulate leukocyte functions

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Barja-Fidalgo C.

2005-01-01

Full Text Available Extracellular matrix proteins and cell adhesion receptors (integrins play essential roles in the regulation of cell adhesion and migration. Interactions of integrins with the extracellular matrix proteins lead to phosphorylation of several intracellular proteins such as focal adhesion kinase, activating different signaling pathways responsible for the regulation of a variety of cell functions, including cytoskeleton mobilization. Once leukocytes are guided to sites of infection, inflammation, or antigen presentation, integrins can participate in the initiation, maintenance, or termination of the immune and inflammatory responses. The modulation of neutrophil activation through integrin-mediated pathways is important in the homeostatic control of the resolution of inflammatory states. In addition, during recirculation, T lymphocyte movement through distinct microenvironments is mediated by integrins, which are critical for cell cycle, differentiation and gene expression. Disintegrins are a family of low-molecular weight, cysteine-rich peptides first identified in snake venom, usually containing an RGD (Arg-Gly-Asp motif, which confers the ability to selectively bind to integrins, inhibiting integrin-related functions in different cell systems. In this review we show that, depending on the cell type and the microenvironment, disintegrins are able to antagonize the effects of integrins or to act agonistically by activating integrin-mediated signaling. Disintegrins have proven useful as tools to improve the understanding of the molecular events regulated by integrin signaling in leukocytes and prototypes in order to design therapies able to interfere with integrin-mediated effects.

Effects of wearing bio-active material coated fabric against γ-irradiation-induced cellular damaged in Sprague-Dawley rats

International Nuclear Information System (INIS)

Kang, Jung Ae; Kim, Hye Rim; Yoon, Sun Hye; Nam, Sang Hyun; Park, Sang Hyun; Jang, Beom Su; Go, Kyung Chan; Yang, Gwang Wung; Rho, Young Hwan; Park, Hyo Suk

2016-01-01

Ionizing radiation causes cellular damage and death through the direct damage and/or indirectly the production of ROS, which induces oxidative stress. This study was designed to evaluate the in vivo radioprotective effects of a bio-active material coated fabric (BMCF) against γ-irradiation-induced cellular damage in Sprague-Dawley (SD) rats. Healthy male SD rats wore bio-active material coated (concentrations in 10% and 30%) fabric for 7 days after 3 Gy of γ-irradiation. Radioprotective effects were evaluated by performing various biochemical assays including spleen and thymus index, WBC count, hepatic damage marker enzymes [aspartate transaminase (AST) and alanine transaminase (ALT)] in plasma, liver antioxidant enzymes, and mitochondrial activity in muscle. Exposure to γ-irradiation resulted in hepatocellular and immune systemic damage. Gamma-irradiation induced decreases in antioxidant enzymes. However, wearing the BMCF-30% decreased significantly AST and ALT activities in plasma. Furthermore, wearing the BMCF-30% increased SOD (superoxide dismutase) and mitochondrial activity. These results suggest that wearing BMCF offers effective radioprotection against γ-irradiation-induced cellular damage in SD rats

Effects of wearing bio-active material coated fabric against γ-irradiation-induced cellular damaged in Sprague-Dawley rats

Energy Technology Data Exchange (ETDEWEB)

Kang, Jung Ae; Kim, Hye Rim; Yoon, Sun Hye; Nam, Sang Hyun; Park, Sang Hyun; Jang, Beom Su [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of); Go, Kyung Chan; Yang, Gwang Wung; Rho, Young Hwan; Park, Hyo Suk [Research and Development Center, VENTEX Co. Ltd., Seoul (Korea, Republic of)

2016-09-15

Ionizing radiation causes cellular damage and death through the direct damage and/or indirectly the production of ROS, which induces oxidative stress. This study was designed to evaluate the in vivo radioprotective effects of a bio-active material coated fabric (BMCF) against γ-irradiation-induced cellular damage in Sprague-Dawley (SD) rats. Healthy male SD rats wore bio-active material coated (concentrations in 10% and 30%) fabric for 7 days after 3 Gy of γ-irradiation. Radioprotective effects were evaluated by performing various biochemical assays including spleen and thymus index, WBC count, hepatic damage marker enzymes [aspartate transaminase (AST) and alanine transaminase (ALT)] in plasma, liver antioxidant enzymes, and mitochondrial activity in muscle. Exposure to γ-irradiation resulted in hepatocellular and immune systemic damage. Gamma-irradiation induced decreases in antioxidant enzymes. However, wearing the BMCF-30% decreased significantly AST and ALT activities in plasma. Furthermore, wearing the BMCF-30% increased SOD (superoxide dismutase) and mitochondrial activity. These results suggest that wearing BMCF offers effective radioprotection against γ-irradiation-induced cellular damage in SD rats.

Separation, Identification, and Bioactivities of the Main Gallotannins of Red Sword Bean (Canavalia gladiata) Coats

Science.gov (United States)

Gan, Ren-You; Kong, Kin-Weng; Li, Hua-Bin; Wu, Kao; Ge, Ying-Ying; Chan, Chak-Lun; Shi, Xian-Ming; Corke, Harold

2018-02-01

The red sword bean (Canavalia gladiata) is an underutilized edible bean cultivated in China. It was previously found to have the highest content of antioxidant polyphenols among 42 edible beans, mainly gallic acid and gallotannins in its red bean coat, an apparently unique characteristic among edible beans. In this study, the main phenolic compounds in red sword bean coats were further separated by Sephadex LH-20 column chromatography, and identified by LC-MS/MS. Furthermore, the FRAP and ABTS antioxidant activities and antibacterial activity (diameter of inhibition zone, DIZ) of main gallotannin-rich fractions were tested. Our results showed that gallotannins of red sword bean coats were mainly comprised of nonogalloyl to hexagalloyl hexosides. Interestingly, tetragalloyl, pentagalloyl, and hexagalloyl hexosides were identified as the main candidates responsible for the red color of the coats. On the other hand, gallotannin-rich fractions exhibited diverse antioxidant and antibacterial activities, and tetragalloyl hexoside overall had the highest free radical scavenging and antibacterial activities. The degree of galloylation did not completely explain the structure-function relationship of gallotannins isolated from red sword bean coats, as there should exist other factors affecting their bioactivities. In conclusion, red sword bean coats are excellent natural sources of gallotannins, and their gallotannin-rich extracts can be utilized as natural antioxidant and antibacterial agents with potential health benefits as well as application in food industry.

Electrophoretic deposition of organic/inorganic composite coatings on metallic substrates for bone replacement applications: mechanisms and development of new bioactive materials based on polysaccharides

OpenAIRE

Cordero Arias, Luis Eduardo

2015-01-01

Regarding the need to improve the usually encountered osteointegration of metallic implants with the surrounding body tissue in bone replacement applications, bioactive organic/inorganic composite coatings on metallic substrates were developed in this work using electrophoretic deposition (EPD) as coating technology. In the present work three polysaccharides, namely alginate, chondroitin sulfate and chitosan were used as the organic part, acting as the matrix of the coating and enabling the c...

In vitro bioactivity of 3D Ti-mesh with bioceramic coatings in simulated body fluid

Directory of Open Access Journals (Sweden)

Wei Yi

2014-09-01

Full Text Available 3D Ti-mesh has been coated with bioceramics under different coating conditions, such as material compositions and micro-porosity, using a dip casting method. Hydroxyapatite (HA, micro-HA particles (HAp, a bioglass (BG and their different mixtures together with polymer additives were used to control HA-coating microstructures. Layered composites with the following coating-to-substrate designs, such as BG/Ti, HA + BG/BG/Ti and HAp + BG/BG/Ti, were fabricated. The bioactivity of these coated composites and the uncoated Ti-mesh substrate was then investigated in a simulated body fluid (SBF. The Ti-mesh substrate and BG/Ti composite did not induce biomimetic apatite deposition when they were immersed in SBF for the selected BG, a pressable dental ceramic, used in this study. After seven days in SBF, an apatite layer was formed on both HA + BG/BG/Ti and HAp + BG/BG/Ti composites. The difference is the apatite layer on the HAp + BG/BG/Ti composite was rougher and contained more micro-pores, while the apatite layer on the HA + BG/BG/Ti composite was dense and smooth. The formation of biomimetic apatite, being more bioresorbable, is favored for bone regeneration.

A peptide affinity column for the identification of integrin alpha IIb-binding proteins.

Science.gov (United States)

Daxecker, Heide; Raab, Markus; Bernard, Elise; Devocelle, Marc; Treumann, Achim; Moran, Niamh

2008-03-01

To understand the regulation of integrin alpha(IIb)beta(3), a critical platelet adhesion molecule, we have developed a peptide affinity chromatography method using the known integrin regulatory motif, LAMWKVGFFKR. Using standard Fmoc chemistry, this peptide was synthesized onto a Toyopearl AF-Amino-650 M resin on a 6-aminohexanoic acid (Ahx) linker. Peptide density was controlled by acetylation of 83% of the Ahx amino groups. Four recombinant human proteins (CIB1, PP1, ICln and RN181), previously identified as binding to this integrin regulatory motif, were specifically retained by the column containing the integrin peptide but not by a column presenting an irrelevant peptide. Hemoglobin, creatine kinase, bovine serum albumin, fibrinogen and alpha-tubulin failed to bind under the chosen conditions. Immunodetection methods confirmed the binding of endogenous platelet proteins, including CIB1, PP1, ICln RN181, AUP-1 and beta3-integrin, from a detergent-free platelet lysate. Thus, we describe a reproducible method that facilitates the reliable extraction of specific integrin-binding proteins from complex biological matrices. This methodology may enable the sensitive and specific identification of proteins that interact with linear, membrane-proximal peptide motifs such as the integrin regulatory motif LAMWKVGFFKR.

Chondroitin Sulfate-E Binds to Both Osteoactivin and Integrin αVβ3 and Inhibits Osteoclast Differentiation.

Science.gov (United States)

Miyazaki, Tatsuya; Miyauchi, Satoshi; Anada, Takahisa; Tawada, Akira; Suzuki, Osamu

2015-10-01

Integrins and their ligands have been suggested to be associated with osteoclast-mediated bone resorption. The present study was designed to investigate whether chondroitin sulfate E (CS-E), which is one of the sulfated glycosaminoglycans (GAGs), is involved in osteoactivin (OA) activity, and osteoclast differentiation. The binding affinity of sulfated GAGs to integrin and its ligand was measured using biotin-labeled CS-E, and the osteoclast differentiation was evaluated by tartrate-resistant acid phosphatase staining and a pit formation assay. CS-E as well as CS-B, synthetic chondroitin polysulfate, and heparin inhibited osteoclast differentiation of bone marrow-derived macrophages. Pre-coating of OA to synthetic calcium phosphate-coated plates enhanced the osteoclastic differentiation of RAW264 cells, and addition of a neutralizing antibody to OA inhibited its differentiation. CS-E bound not only to OA, fibronectin, and vitronectin, but also to its receptor integrin αVβ3, and inhibited the direct binding of OA to integrin αVβ3. Furthermore, CS-E blocked the binding of OA to cells and inhibited OA-induced osteoclastic differentiation. On the other hand, heparinase treatment of RAW264 cells inhibited osteoclastic differentiation. Since binding of OA to the cells was inhibited by the presence of heparan sulfate or heparinase treatment of cells, heparan sulfate proteoglycan (HSPG) was also considered to be an OA receptor. Taken together, the present results suggest that CS-E is capable of inhibiting OA-induced osteoclast differentiation by blocking the interaction of OA to integrin αVβ3 and HSPG. © 2015 Wiley Periodicals, Inc.

Surface coated polyurethane with improved bioactivity and cytocompatability

CSIR Research Space (South Africa)

Chetty, AS

2006-02-01

Full Text Available Polyurethane (PU) may be suitable for various implant applications; however, it lacks bioactivity. Bioactivity allows for direct tissue attachment at the bio- interface, enabling implant fixation while preventing fibrous encapsulation. To impart...

Bioactive glass-ceramic coating for enhancing the in vitro corrosion resistance of biodegradable Mg alloy

Energy Technology Data Exchange (ETDEWEB)

Ye Xinyu [Key Laboratory for Advanced Ceramics and Machining Technology of Ministry of Education, Tianjin University, Tianjin 300072 (China); Cai Shu, E-mail: caishu@tju.edu.cn [Key Laboratory for Advanced Ceramics and Machining Technology of Ministry of Education, Tianjin University, Tianjin 300072 (China); Dou Ying [Key Laboratory for Advanced Ceramics and Machining Technology of Ministry of Education, Tianjin University, Tianjin 300072 (China); Xu Guohua [Shanghai Changzheng Hospital, Shanghai 200003 (China); Huang Kai; Ren Mengguo; Wang Xuexin [Key Laboratory for Advanced Ceramics and Machining Technology of Ministry of Education, Tianjin University, Tianjin 300072 (China)

2012-10-15

Highlights: Black-Right-Pointing-Pointer Sol-gel derived 45S5 glass-ceramic coating was prepared on Mg alloy substrate. Black-Right-Pointing-Pointer The corrosion resistance of glass-ceramic coated Mg alloy was markedly improved. Black-Right-Pointing-Pointer The corrosion behavior of the coated sample varied due to the cracking of coating. - Abstract: In this work, a bioactive 45S5 glass-ceramic coating was synthesized on magnesium (Mg) alloy substrate by using a sol-gel dip-coating method, to improve the initial corrosion resistance of AZ31 Mg alloy. The surface morphology and phase composition of the glass-ceramic coating were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The coating composed of amorphous phase and crystalline phase Na{sub 2}Ca{sub 2}Si{sub 3}O{sub 9}, with the thickness of {approx}1.0 {mu}m, exhibited a uniform and crack-free surface morphology. The corrosion behavior of the uncoated and coated Mg alloy substrates was investigated by the electrochemical measurements and immersion tests in simulated body fluid (SBF). Potentiodynamic polarization tests recorded an increase of potential (E{sub corr}) form -1.60 V to -1.48 V, and a reduction of corrosion current density (i{sub corr}) from 4.48 {mu}A cm{sup -2} to 0.16 {mu}A cm{sup -2}, due to the protection provided by the glass-ceramic coating. Immersion tests also showed the markedly improved corrosion resistance of the coated sample over the immersion period of 7 days. Moreover, after 14 days of immersion in SBF, the corrosion resistance of the coated sample declined due to the cracking of the glass-ceramic coating, which was confirmed by electrochemical impedance spectroscopy (EIS) analysis. The results suggested that the 45S5 glass-ceramic coated Mg alloy could provide a suitable corrosion behavior for use as degradable implants.

Cardiac integrins the ties that bind.

Science.gov (United States)

Simpson, D G; Reaves, T A; Shih, D T; Burgess, W; Borg, T K; Terracio, L

1998-01-01

An elaborate series of morphogenetic events must be precisely coordinated during development to promote the formation of the elaborate three-dimensional structure of the normal heart. In this study we focus on discussing how interconnections between the cardiac myocyte and its surrounding environment regulate cardiac form and function. In vitro experiments from our laboratories provide direct evidence that cardiac cell shape is regulated by a dynamic interaction between constituents of the extracellular matrix (ECM) and by specific members of the integrin family of matrix receptors. Our data indicates that phenotypic information is stored in the tertiary structure and chemical identity of the ECM. This information appears to be actively communicated and transduced by the α1β1 integrin molecule into an intracellular signal that regulates cardiac cell shape and myofibrillar organization. In this study we have assessed the phenotypic consequences of suppressing the expression and accumulation of the α1 integrin molecule in aligned cultures of cardiac myocytes. In related experiments we have examined how the overexpression of α2 and α5 integrin, integrins normally not present or present at very low copy number on the cell surface of neonatal cardiac myocytes, affect cardiac protein metabolism. We also consider how biochemical signals and the mechanical signals mediated by the integrins may converge on common intracellular signaling pathways in the heart. Experiments with the whole embryo culture system indicate that angiotensin II, a peptide that carries information concerning cardiac load, plays a role in controling cardiac looping and the proliferation of myofibrils during development.

Search for specific biomarkers of IFNβ bioactivity in patients with multiple sclerosis.

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Sunny Malhotra

Full Text Available Myxovirus A (MxA, a protein encoded by the MX1 gene with antiviral activity, has proven to be a sensitive measure of IFNβ bioactivity in multiple sclerosis (MS. However, the use of MxA as a biomarker of IFNβ bioactivity has been criticized for the lack of evidence of its role on disease pathogenesis and the clinical response to IFNβ. Here, we aimed to identify specific biomarkers of IFNβ bioactivity in order to compare their gene expression induction by type I IFNs with the MxA, and to investigate their potential role in MS pathogenesis. Gene expression microarrays were performed in PBMC from MS patients who developed neutralizing antibodies (NAB to IFNβ at 12 and/or 24 months of treatment and patients who remained NAB negative. Nine genes followed patterns in gene expression over time similar to the MX1, which was considered the gold standard gene, and were selected for further experiments: IFI6, IFI27, IFI44L, IFIT1, HERC5, LY6E, RSAD2, SIGLEC1, and USP18. In vitro experiments in PBMC from healthy controls revealed specific induction of selected biomarkers by IFNβ but not IFNγ, and several markers, in particular USP18 and HERC5, were shown to be significantly induced at lower IFNβ concentrations and more selective than the MX1 as biomarkers of IFNβ bioactivity. In addition, USP18 expression was deficient in MS patients compared with healthy controls (p = 0.0004. We propose specific biomarkers that may be considered in addition to the MxA to evaluate IFNβ bioactivity, and to further explore their implication in MS pathogenesis.

Age-specific function of α5β1 integrin in microglial migration during early colonization of the developing mouse cortex.

Science.gov (United States)

Smolders, Sophie Marie-Thérèse; Swinnen, Nina; Kessels, Sofie; Arnauts, Kaline; Smolders, Silke; Le Bras, Barbara; Rigo, Jean-Michel; Legendre, Pascal; Brône, Bert

2017-07-01

Microglia, the immune cells of the central nervous system, take part in brain development and homeostasis. They derive from primitive myeloid progenitors that originate in the yolk sac and colonize the brain mainly through intensive migration. During development, microglial migration speed declines which suggests that their interaction with the microenvironment changes. However, the matrix-cell interactions allowing dispersion within the parenchyma are unknown. Therefore, we aimed to better characterize the migration behavior and to assess the role of matrix-integrin interactions during microglial migration in the embryonic brain ex vivo. We focused on microglia-fibronectin interactions mediated through the fibronectin receptor α5β1 integrin because in vitro work indirectly suggested a role for this ligand-receptor pair. Using 2-photon time-lapse microscopy on acute ex vivo embryonic brain slices, we found that migration occurs in a saltatory pattern and is developmentally regulated. Most importantly, there is an age-specific function of the α5β1 integrin during microglial cortex colonization. At embryonic day (E) 13.5, α5β1 facilitates migration while from E15.5, it inhibits migration. These results indicate a developmentally regulated function of α5β1 integrin in microglial migration during colonization of the embryonic brain. © 2017 Wiley Periodicals, Inc.

Bio-active molecules modified surfaces enhanced mesenchymal stem cell adhesion and proliferation

International Nuclear Information System (INIS)

Mobasseri, Rezvan; Tian, Lingling; Soleimani, Masoud; Ramakrishna, Seeram; Naderi-Manesh, Hossein

2017-01-01

Surface modification of the substrate as a component of in vitro cell culture and tissue engineering, using bio-active molecules including extracellular matrix (ECM) proteins or peptides derived ECM proteins can modulate the surface properties and thereby induce the desired signaling pathways in cells. The aim of this study was to evaluate the behavior of human bone marrow mesenchymal stem cells (hBM-MSCs) on glass substrates modified with fibronectin (Fn), collagen (Coll), RGD peptides (RGD) and designed peptide (R-pept) as bio-active molecules. The glass coverslips were coated with fibronectin, collagen, RGD peptide and R-peptide. Bone marrow mesenchymal stem cells were cultured on different substrates and the adhesion behavior in early incubation times was investigated using scanning electron microscopy (SEM) and confocal microscopy. The MTT assay was performed to evaluate the effect of different bio-active molecules on MSCs proliferation rate during 24 and 72 h. Formation of filopodia and focal adhesion (FA) complexes, two steps of cell adhesion process, were observed in MSCs cultured on bio-active molecules modified coverslips, specifically in Fn coated and R-pept coated groups. SEM image showed well adhesion pattern for MSCs cultured on Fn and R-pept after 2 h incubation, while the shape of cells cultured on Coll and RGD substrates indicated that they might experience stress condition in early hours of culture. Investigation of adhesion behavior, as well as proliferation pattern, suggests R-peptide as a promising bio-active molecule to be used for surface modification of substrate in supporting and inducing cell adhesion and proliferation. - Highlights: • Bioactive molecules modified surface is a strategy to design biomimicry scaffold. • Bi-functional Tat-derived peptide (R-pept) enhanced MSCs adhesion and proliferation. • R-pept showed similar influences to fibronectin on FA formation and attachment.

3D analysis of thermal and stress evolution during laser cladding of bioactive glass coatings.

Science.gov (United States)

Krzyzanowski, Michal; Bajda, Szymon; Liu, Yijun; Triantaphyllou, Andrew; Mark Rainforth, W; Glendenning, Malcolm

2016-06-01

Thermal and strain-stress transient fields during laser cladding of bioactive glass coatings on the Ti6Al4V alloy basement were numerically calculated and analysed. Conditions leading to micro-cracking susceptibility of the coating have been investigated using the finite element based modelling supported by experimental results of microscopic investigation of the sample coatings. Consecutive temperature and stress peaks are developed within the cladded material as a result of the laser beam moving along the complex trajectory, which can lead to micro-cracking. The preheated to 500°C base plate allowed for decrease of the laser power and lowering of the cooling speed between the consecutive temperature peaks contributing in such way to achievement of lower cracking susceptibility. The cooling rate during cladding of the second and the third layer was lower than during cladding of the first one, in such way, contributing towards improvement of cracking resistance of the subsequent layers due to progressive accumulation of heat over the process. Copyright © 2016 Elsevier Ltd. All rights reserved.

The role of mechanical force and ROS in integrin-dependent signals.

Directory of Open Access Journals (Sweden)

Kathrin S Zeller

Full Text Available Cells are exposed to several types of integrin stimuli, which generate responses generally referred to as "integrin signals", but the specific responses to different integrin stimuli are poorly defined. In this study, signals induced by integrin ligation during cell attachment, mechanical force from intracellular contraction, or cell stretching by external force were compared. The elevated phosphorylation levels of several proteins during the early phase of cell attachment and spreading of fibroblast cell lines were not affected by inhibition of ROCK and myosin II activity, i.e. the reactions occurred independently of intracellular contractile force acting on the adhesion sites. The contraction-independent phosphorylation sites included ERK1/2 T202/Y204, AKT S473, p130CAS Y410, and cofilin S3. In contrast to cell attachment, cyclic stretching of the adherent cells induced a robust phosphorylation only of ERK1/2 and the phosphorylation levels of the other investigated proteins were not or only moderately affected by stretching. No major differences between signaling via α5β1 or αvβ3 integrins were detected. The importance of mitochondrial ROS for the integrin-induced signaling pathways was investigated using rotenone, a specific inhibitor of complex I in the respiratory chain. While rotenone only moderately reduced ATP levels and hardly affected the signals induced by cyclic cell stretching, it abolished the activation of AKT and reduced the actin polymerization rate in response to attachment in both cell lines. In contrast, scavenging of extracellular ROS with catalase or the vitamin C analog Asc-2P did not significantly influence the attachment-derived signaling, but caused a selective and pronounced enhancement of ERK1/2 phosphorylation in response to stretching. In conclusion, the results showed that "integrin signals" are composed of separate sets of reactions triggered by different types of integrin stimulation. Mitochondrial ROS and

Antibacterial polylactic acid/chitosan nanofibers decorated with bioactive glass

Energy Technology Data Exchange (ETDEWEB)

Goh, Yi-fan; Akram, Muhammad; Alshemary, Ammarz [Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Hussain, Rafaqat, E-mail: rafaqat@comsats.edu.pk [Department of Physics, COMSATS Institute of Information Technology, Islamabad (Pakistan)

2016-11-30

Highlights: • PLA/Chitosan nanofibers were coated with functional bioglass. • Polymer/ceramic composite fibers exhibited good in-vitro bioactivity. • Nanofibers coated with Ag doped bioglass exhibited good antibacterial activity. - Abstract: In this study, we have presented the structural and in vitro characterization of electrospun polylactic acid (PLA)/Chitosan nanofibers coated with cerium, copper or silver doped bioactive glasses (CeBG/CuBG/AgBG). Bead-free, smooth surfaced nanofibers were successfully prepared by using electrospinning technique. The nanocomposite fibers were obtained using a facile dip-coating method, their antibacterial activities against E. coliE. coli (ATCC 25922 strains) were measured by the disk diffusion method after 24 h of incubation at 37 °C. CeBG and CuBG decorated PLA/Chitosan nanofibers did not develop an inhibition zone against the bacteria. On the other hand, nanofibers coated with AgBG developed an inhibition zone against the bacteria. The as-prepared nanocomposite fibers were immersed in SBF for 1, 3 and 7 days in Simulated Body Fluid (SBF) for evaluation of in vitro bioactivity. All samples induced the formation of crystallites with roughly ruffled morphology and the pores of fibers were covered with the extensive growth of crystallites. Energy Dispersive X-ray (EDX) composition analysis showed that the crystallites possessed Ca/P ratio close to 1.67, confirming the good in-vitro bioactivity of the fibers.

Application of magnetron sputtering for producing bioactive ceramic ...

Indian Academy of Sciences (India)

Wintec

time is to coat the metal surface with a bioactive material that can promote rapid ... metal implant and the surrounding tissue because of their close similarity to the ... good control of coating composition, a good adhesion of the coating to the ...

Integrin β1 regulates leiomyoma cytoskeletal integrity and growth

Science.gov (United States)

Malik, Minnie; Segars, James; Catherino, William H.

2014-01-01

Uterine leiomyomas are characterized by an excessive extracellular matrix, increased mechanical stress, and increased active RhoA. Previously, we observed that mechanical signaling was attenuated in leiomyoma, but the mechanisms responsible remain unclear. Integrins, especially integrin β1, are transmembrane adhesion receptors that couple extracellular matrix stresses to the intracellular cytoskeleton to influence cell proliferation and differentiation. Here we characterized integrin and laminin to signaling in leiomyoma cells. We observed a 2.25 ± 0.32 fold increased expression of integrin β1 in leiomyoma cells, compared to myometrial cells. Antibody-mediated inhibition of integrin β1 led to significant growth inhibition in leiomyoma cells and a loss of cytoskeletal integrity. Specifically, polymerization of actin filaments and formation of focal adhesions were reduced by inhibition of integrin p1. Inhibition of integrin β1 in leiomyoma cells led to 0.81 ± 0.02 fold decrease in active RhoA, and resembled levels found in serum-starved cells. Likewise, inhibition of integrin β1 was accompanied by a decrease in phospho-ERK. Compared to myometrial cells, leiomyoma cells demonstrated increased expression of integrin α6 subunit to laminin receptor (1.91 ± 0.11 fold), and increased expression of laminin 5α (1.52±0.02), laminin 5β (3.06±0.92), and laminin 5γ (1.66 ± 0.06). Of note, leiomyoma cells grown on laminin matrix appear to realign themselves. Taken together, the findings reveal that the attenuated mechanical signaling in leiomyoma cells is accompanied by an increased expression and a dependence on integrin β1 signaling in leiomyoma cells, compared to myometrial cells. PMID:23023061

Integrin α5β1, the Fibronectin Receptor, as a Pertinent Therapeutic Target in Solid Tumors

Energy Technology Data Exchange (ETDEWEB)

Schaffner, Florence; Ray, Anne Marie; Dontenwill, Monique, E-mail: monique.dontenwill@unistra.fr [UMR 7213 CNRS, Laboratoire de Biophotonique et Pharmacologie, Tumoral signaling and therapeutic targets, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch (France)

2013-01-15

Integrins are transmembrane heterodimeric proteins sensing the cell microenvironment and modulating numerous signalling pathways. Changes in integrin expression between normal and tumoral cells support involvement of specific integrins in tumor progression and aggressiveness. This review highlights the current knowledge about α5β1 integrin, also called the fibronectin receptor, in solid tumors. We summarize data showing that α5β1 integrin is a pertinent therapeutic target expressed by tumoral neovessels and tumoral cells. Although mainly evaluated in preclinical models, α5β1 integrin merits interest in particular in colon, breast, ovarian, lung and brain tumors where its overexpression is associated with a poor prognosis for patients. Specific α5β1 integrin antagonists will be listed that may represent new potential therapeutic agents to fight defined subpopulations of particularly aggressive tumors.

Fabrication and design of bioactive agent coated, highly-aligned electrospun matrices for nerve tissue engineering: Preparation, characterization and application

Science.gov (United States)

Lee, Sang Jin; Heo, Min; Lee, Donghyun; Heo, Dong Nyoung; Lim, Ho-Nam; Kwon, Il Keun

2017-12-01

In this study, we designed highly-aligned thermoplastic polycarbonate urethane (PCU) fibrous scaffolds coated with bioactive compounds, such as Poly-L-Lysine (PLL) and Poly-L-Ornithine (PLO), to enhance cellular adhesion and directivity. These products were characterized by scanning electron microscope (SEM) analysis which demonstrated that highly aligned fiber strands were formed without beads when coated onto a mandrel rotating at 1800 rpm. During in vitro cell test, PLO-coated, aligned PCU scaffolds were found to have significantly higher proliferation rates than PLL coated and bare PCU scaffolds. Interestingly, dental pulp stem cells (DPSCs) were observed to stretch along the longitudinal axis parallel to the cell direction on highly aligned scaffolds. These results clearly confirm that our strategy may suggest a useful paradigm by inducing neural tissue repair as a means to remodeling and healing of tissue for restorative procedures in neural tissue engineering.

Preliminary research on a novel bioactive silicon doped calcium phosphate coating on AZ31 magnesium alloy via electrodeposition

Energy Technology Data Exchange (ETDEWEB)

Qiu, Xun [Department of Materials Science and Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Institute of Metal Research, Chinese Academy of Science, Shenyang 110016 (China); Wan, Peng, E-mail: pwan@imr.ac.cn [Institute of Metal Research, Chinese Academy of Science, Shenyang 110016 (China); Tan, LiLi [Institute of Metal Research, Chinese Academy of Science, Shenyang 110016 (China); Fan, XinMin [Department of Materials Science and Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Ke [Institute of Metal Research, Chinese Academy of Science, Shenyang 110016 (China)

2014-03-01

A silicon doped calcium phosphate coating was obtained successfully on AZ31 alloy substrate via pulse electrodeposition. A novel dual-layer structure was observed with a porous lamellar-like and outer block-like apatite layer. In vitro immersion tests were adopted in simulated body fluid within 28 days of immersion. Slow degradation rate obtained from weight loss was observed for the Si-doped Ca–P coating, which was also consistent with the results of electrochemical experiments showing an enhanced corrosion resistance for the coating. Further formation of an apatite-like layer on the surface after immersion proved better integrity and biomineralization performance of the coating. Biological characterization was carried out for viability, proliferation and differentiation of MG63 osteoblast-like cells. The coating showed a good cell growth and an enhanced cell proliferation. Moreover, an increased activity of osteogenic marker ALP was found. All the results demonstrated that the Si-doped calcium phosphate was perspective to be used as a coating for magnesium alloy implants to control the degradation rate and enhance the bioactivity, which would facilitate the rapidity of bone tissue repair. - Highlights: • A Si-doped calcium phosphate coating was achieved via pulse ED on AZ31 alloy. • The coating was composed of a porous lamellar-like layer and outer block-like apatite. • The coating showed slow degradation rate and better biomineralization property. • The coating improved cell proliferation and activity of osteogenic marker ALP.

Preliminary research on a novel bioactive silicon doped calcium phosphate coating on AZ31 magnesium alloy via electrodeposition

International Nuclear Information System (INIS)

Qiu, Xun; Wan, Peng; Tan, LiLi; Fan, XinMin; Yang, Ke

2014-01-01

A silicon doped calcium phosphate coating was obtained successfully on AZ31 alloy substrate via pulse electrodeposition. A novel dual-layer structure was observed with a porous lamellar-like and outer block-like apatite layer. In vitro immersion tests were adopted in simulated body fluid within 28 days of immersion. Slow degradation rate obtained from weight loss was observed for the Si-doped Ca–P coating, which was also consistent with the results of electrochemical experiments showing an enhanced corrosion resistance for the coating. Further formation of an apatite-like layer on the surface after immersion proved better integrity and biomineralization performance of the coating. Biological characterization was carried out for viability, proliferation and differentiation of MG63 osteoblast-like cells. The coating showed a good cell growth and an enhanced cell proliferation. Moreover, an increased activity of osteogenic marker ALP was found. All the results demonstrated that the Si-doped calcium phosphate was perspective to be used as a coating for magnesium alloy implants to control the degradation rate and enhance the bioactivity, which would facilitate the rapidity of bone tissue repair. - Highlights: • A Si-doped calcium phosphate coating was achieved via pulse ED on AZ31 alloy. • The coating was composed of a porous lamellar-like layer and outer block-like apatite. • The coating showed slow degradation rate and better biomineralization property. • The coating improved cell proliferation and activity of osteogenic marker ALP

Structure, composition and morphology of bioactive titanate layer on porous titanium surfaces

Science.gov (United States)

Li, Jinshan; Wang, Xiaohua; Hu, Rui; Kou, Hongchao

2014-07-01

A bioactive coating was produced on pore surfaces of porous titanium samples by an amendatory alkali-heat treatment method. Porous titanium was prepared by powder metallurgy and its porosity and average size were 45% and 135 μm, respectively. Coating morphology, coating structure and phase constituents were examined by SEM, XPS and XRD. It was found that a micro-network structure with sizes of cells, and redundant Ca ion was detected in the titanate layer. The concentration distribution of Ti, O, Ca and Na in the coating showed a compositional gradient from the intermediate layer toward the outer surface. These compositional gradients indicate that the coating bonded to Ti substrate without a distinct interface. After immersion into the SBF solution for 3 days, a bone-like carbonate-hydroxylapatite showing a good biocompatibility was detected on the coating surface. And the redundant Ca advanced the bioactivity of the coating. Thus, the present modification is expected to allow the use of the bioactive porous titanium as artificial bones even under load-bearing conditions.

The newcomer in the integrin family: Integrin α9 in biology and cancer

DEFF Research Database (Denmark)

Høye, Anette Melissa; Couchman, John Robert; Wewer, Ulla M.

2012-01-01

Integrins are heterodimeric transmembrane receptors regulating cell-cell and cell-extracellular matrix interactions. Of the 24 integrin heterodimers identified in humans, a9ß1 integrin is one of the least studied. a9, together with a4, comprise a more recent evolutionary sub-family of integrins...... of cell types, interacts with many ligands for example fibronectin, tenascin-C and ADAM12, and has been shown to have important functions in processes such as cell adhesion and migration, lung development, lymphatic and venous valve development, and in wound healing. This has sparked an interest...

Beta1 integrin-mediated adhesion signalling is essential for epidermal progenitor cell expansion

DEFF Research Database (Denmark)

Piwko-Czuchra, Aleksandra; Koegel, Heidi; Meyer, Hannelore

2009-01-01

BACKGROUND: There is a major discrepancy between the in vitro and in vivo results regarding the role of beta1 integrins in the maintenance of epidermal stem/progenitor cells. Studies of mice with skin-specific ablation of beta1 integrins suggested that epidermis can form and be maintained in thei...... of increased keratinocyte proliferation such as wound healing. CONCLUSIONS/SIGNIFICANCE: These data demonstrate that expression of beta1 integrins is critically important for the expansion of epidermal progenitor cells to maintain epidermal homeostasis....... that developed similar, but less severe defects than mice with beta1-deficient keratinocytes. Surprisingly we found that upon aging these abnormalities attenuated due to a rapid expansion of cells, which escaped or compensated for the down-regulation of beta1 integrin expression. A similar phenomenon...... was observed in aged mice with a complete, skin-specific ablation of the beta1 integrin gene, where cells that escaped Cre-mediated recombination repopulated the mutant skin in a very short time period. The expansion of beta1 integrin expressing keratinocytes was even further accelerated in situations...

Elucidating the role of select cytoplasmic proteins in altering diffusion of integrin receptors.

Science.gov (United States)

Sander, Suzanne; Arora, Neha; Smith, Emily A

2012-06-01

Cytoplasmic proteins that affect integrin diffusion in the cell membrane are identified using a combination of fluorescence recovery after photobleaching (FRAP) and RNA interference. Integrin receptors are essential for many cellular events, and alterations in lateral diffusion are one mechanism for modulating their function. In cells expressing native cytoplasmic protein concentrations and spread on a slide containing integrin extracellular ligand, 45 ± 2% of the integrin is mobile with a time-dependent 5.2 ± 0.9 × 10(-9) cm(2)/s diffusion coefficient at 1 s. The time exponent is 0.90 ± 0.07, indicating integrin diffusion moderately slows at longer times. The role of a specific cytoplasmic protein in altering integrin diffusion is revealed through changes in the FRAP curve after reducing the cytoplasmic protein's expression. Decreased expression of cytoplasmic proteins rhea, focal adhesion kinase (FAK), or steamer duck decreases the integrin mobile fraction. For rhea and FAK, there is a concomitant shift to Brownian (i.e., time-independent) diffusion at reduced concentrations of these proteins. In contrast, when the expression of actin 42A, dreadlocks, paxillin, integrin-linked kinase (ILK), or vinculin is reduced, integrin diffusion generally becomes more constrained with an increase in the integrin mobile fraction. This same change in integrin diffusion is measured in the absence of integrin extracellular ligand. The results indicate breaking the extracellular ligand-integrin-cytoskeletal linkage alters integrin diffusion properties, and, in most cases, there is no correlation between integrin and lipid diffusion properties.

The role of alpha3beta1 integrin in determining the supramolecular organization of laminin-5 in the extracellular matrix of keratinocytes.

Science.gov (United States)

deHart, Gregory W; Healy, Kevin E; Jones, Jonathan C R

2003-02-01

Analyses of mice with targeted deletions in the genes for alpha3 and beta1 integrin suggest that the alpha3beta1 integrin heterodimer likely determines the organization of the extracellular matrix within the basement membrane of skin. Here we tested this hypothesis using keratinocytes derived from alpha3 integrin-null mice. We have compared the organizational state of laminin-5, a ligand of alpha3beta1 integrin, in the matrix of wild-type keratinocytes with that of laminin-5 in the matrix of alpha3 integrin-null cells. Laminin-5 distributes diffusely in arc structures in the matrix of wild-type mouse keratinocytes, whereas laminin-5 is organized into linear, spike-like arrays by the alpha3 integrin-null cells. The fact that alpha3 integrin-null cells are deficient in their ability to assemble a proper laminin-5 matrix is also shown by their failure to remodel laminin-5 when plated onto surfaces coated with purified laminin-5 protein. In sharp contrast, wild-type keratinocytes organize exogenously added laminin-5 into discrete ring-like organizations. These findings led us next to assess whether differences in laminin-5 organization in the matrix of the wild-type and alpha3 integrin-null cells impact cell behavior. Our results indicate that alpha3 integrin-null cells are more motile than their wild-type counterparts and leave extensive trails of laminin-5 over the surface on which they move. Moreover, HEK 293 cells migrate significantly more on the laminin-5-rich matrix derived from the alpha3 integrin-null cells than on the wild-type keratinocyte laminin-5 matrix. In addition, alpha3 integrin-null cells show low strength of adhesion to surfaces coated with purified laminin-5 compared to wild-type cells although both the wild type and the alpha3 integrin-null keratinocytes adhere equally strongly to laminin-5 that has been organized into arrays by other epithelial cells. These data suggest: (1) that alpha3beta1 integrin plays an important role in determining the

Activation of c-Raf-1 kinase signal transduction pathway in alpha(7) integrin-deficient mice.

Science.gov (United States)

Saher, G; Hildt, E

1999-09-24

Integrin alpha(7)-deficient mice develop a novel form of muscular dystrophy. Here we report that deficiency of alpha(7) integrin causes an activation of the c-Raf-1/mitogen-activated protein (MAP) 2 kinase signal transduction pathway in muscle cells. The observed activation of c-Raf-1/MAP2 kinases is a specific effect, because the alpha(7) integrin deficiency does not cause unspecific stress as determined by measurement of the Hsp72/73 level and activity of the JNK2 kinase. Because an increased level of activated FAK was found in muscle of alpha(7) integrin-deficient mice, the activation of c-Raf-1 kinase is triggered most likely by an integrin-dependent pathway. In accordance with this, in the integrin alpha(7)-deficient mice, part of the integrin beta(1D) variant in muscle is replaced by the beta(1A) variant, which permits the FAK activation. A recent report describes that integrin activity can be down-modulated by the c-Raf-1/MAP2 kinase pathway. Specific activation of the c-Raf-1/MAP2 kinases by cell-permeable peptides in skeletal muscle of rabbits causes degeneration of muscle fibers. Therefore, we conclude that in alpha(7) integrin-deficient mice, the continuous activation of c-Raf-1 kinase causes a permanent reduction of integrin activity diminishing integrin-dependent cell-matrix interactions and thereby contributing to the development of the dystrophic phenotype.

Diverse roles of integrin receptors in articular cartilage.

Science.gov (United States)

Shakibaei, M; Csaki, C; Mobasheri, A

2008-01-01

Integrins are heterodimeric integral membrane proteins made up of alpha and beta subunits. At least eighteen alpha and eight beta subunit genes have been described in mammals. Integrin family members are plasma membrane receptors involved in cell adhesion and active as intra- and extracellular signalling molecules in a variety of processes including embryogenesis, hemostasis, tissue repair, immune response and metastatic spread of tumour cells. Integrin beta 1 (beta1-integrin), the protein encoded by the ITGB1 gene (also known as CD29 and VLAB), is a multi-functional protein involved in cell-matrix adhesion, cell signalling, cellular defense, cell adhesion, protein binding, protein heterodimerisation and receptor-mediated activity. It is highly expressed in the human body (17.4 times higher than the average gene in the last updated revision of the human genome). The extracellular matrix (ECM) of articular cartilage is a unique environment. Interactions between chondrocytes and the ECM regulate many biological processes important to homeostasis and repair of articular cartilage, including cell attachment, growth, differentiation and survival. The beta1-integrin family of cell surface receptors appears to play a major role in mediating cell-matrix interactions that are important in regulating these fundamental processes. Chondrocyte mechanoreceptors have been proposed to incorporate beta1-integrins and mechanosensitive ion channels which link with key ECM, cytoskeletal and signalling proteins to maintain the chondrocyte phenotype, prevent chondrocyte apoptosis and regulate chondrocyte-specific gene expression. This review focuses on the expression and function of beta1-integrins in articular chondrocytes, its role in the unique biology of these cells and its distribution in cartilage.

Preparation, characterization and in vitro response of bioactive coatings on polyether ether ketone.

Science.gov (United States)

Durham, John W; Allen, Matthew J; Rabiei, Afsaneh

2017-04-01

Polyether ether ketone (PEEK) is a highly heat-resistant thermoplastic with excellent strength and elastic modulus similar to human bone, making it an attractive material for orthopedic implants. However, the hydrophobic surface of PEEK implants induces fibrous encapsulation which is unfavorable for stable implant anchorage. In this study, PEEK was coated via ion-beam-assisted deposition (IBAD) using a two-layer design of yttria-stabilized zirconia (YSZ) as a heat-protection layer, and hydroxyapatite (HA) as a top layer to improve osseointegration. Microstructural analysis of the coatings showed a dense, uniform columnar grain structure in the YSZ layer and no delamination from the substrate. The HA layer was found to be amorphous and free of porosities in its as-deposited state. Subsequent heat treatment via microwave energy followed by autoclaving crystallized the HA layer, confirmed by SEM and XRD analysis. An in vitro study using MC3T3 preosteoblast cells showed improved bioactivity in heat-treated sample groups. Cell proliferation, differentiation, and mineralization were analyzed by MTT assay and DNA content, osteocalcin expression, and Alizarin Red S (AR-S) content, respectively. Initial cell growth was increased, and osteogenic maturation and mineralization were accelerated most on coatings that underwent a combined microwave and autoclave heat treatment process as compared to uncoated PEEK and amorphous HA surfaces. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 560-567, 2017. © 2015 Wiley Periodicals, Inc.

Corrosion resistance and in-vitro bioactivity of BaO containing Na2O-CaO-P2O5 phosphate glass-ceramic coating prepared on 316 L, duplex stainless steel 2205 and Ti6Al4V

Science.gov (United States)

Edathazhe, Akhila B.; Shashikala, H. D.

2018-03-01

The phosphate glass with composition 11Na2O-15BaO-29CaO-45P2O5 was coated on biomedical implant materials such as stainless steel 316 L, duplex stainless steel (DSS) 2205 and Ti6Al4V alloy by thermal enamelling method. The structural properties and composition of glass coated substrates were studied by x-ray diffraction (XRD), Scanning electron microscopy (SEM) and Energy dispersive x-ray spectroscopy (EDS) analysis. The coatings were partially crystalline in nature with porous structure and pore size varied from micro to nanometer range. The polarization curve was obtained for uncoated and coated substrates from electrochemical corrosion test which was conducted at 37 °C in Hank’s balanced salt solution (HBSS). The corrosion resistance of 316 L substrate increased after coating, whereas it decreased in case of DSS 2205 and Ti6Al4V. The XRD and SEM/EDS studies indicated the bioactive hydroxyapatite (HAp) layer formation on all the coated surfaces after electrochemical corrosion test, which improved the corrosion resistance. The observed electrochemical corrosion behavior can be explained based on protective HAp layer formation, composition and diffusion of ions on glass coated surfaces. The in-vitro bioactivity test was carried out at 37 °C in HBS solution for 14 days under static conditions for uncoated and coated substrates. pH and ion release rate measurements from the coated samples were conducted to substantiate the electrochemical corrosion test. The lower ion release rates of Na+ and Ca2+ from coated 316 L supported its higher electrochemical corrosion resistance among coated samples. Among the uncoated substrates, DSS showed higher electrochemical corrosion resistance. Amorphous calcium-phosphate (ACP) layer formation on all the coated substrates after in-vitro bioactivity test was confirmed by XRD, SEM/EDS and ion release measurements. The present work is a comparative study of corrosion resistance and bioactivity of glass coated and uncoated

Complementary roles of KCa3.1 channels and β1-integrin during alveolar epithelial repair.

Science.gov (United States)

Girault, Alban; Chebli, Jasmine; Privé, Anik; Trinh, Nguyen Thu Ngan; Maillé, Emilie; Grygorczyk, Ryszard; Brochiero, Emmanuelle

2015-09-04

Extensive alveolar epithelial injury and remodelling is a common feature of acute lung injury and acute respiratory distress syndrome (ARDS) and it has been established that epithelial regeneration, and secondary lung oedema resorption, is crucial for ARDS resolution. Much evidence indicates that K(+) channels are regulating epithelial repair processes; however, involvement of the KCa3.1 channels in alveolar repair has never been investigated before. Wound-healing assays demonstrated that the repair rates were increased in primary rat alveolar cell monolayers grown on a fibronectin matrix compared to non-coated supports, whereas an anti-β1-integrin antibody reduced it. KCa3.1 inhibition/silencing impaired the fibronectin-stimulated wound-healing rates, as well as cell migration and proliferation, but had no effect in the absence of coating. We then evaluated a putative relationship between KCa3.1 channel and the migratory machinery protein β1-integrin, which is activated by fibronectin. Co-immunoprecipitation and immunofluorescence experiments indicated a link between the two proteins and revealed their cellular co-distribution. In addition, we demonstrated that KCa3.1 channel and β1-integrin membrane expressions were increased on a fibronectin matrix. We also showed increased intracellular calcium concentrations as well as enhanced expression of TRPC4, a voltage-independent calcium channel belonging to the large TRP channel family, on a fibronectin matrix. Finally, wound-healing assays showed additive effects of KCa3.1 and TRPC4 inhibitors on alveolar epithelial repair. Taken together, our data demonstrate for the first time complementary roles of KCa3.1 and TRPC4 channels with extracellular matrix and β1-integrin in the regulation of alveolar repair processes.

Mg substituted apatite coating from alkali conversion of acidic calcium phosphate

Energy Technology Data Exchange (ETDEWEB)

Navarro da Rocha, Daniel, E-mail: dnr.navarro@gmail.com [Military Institute of Engineering-IME, Pça. Gen. Tiburcio, 80, P. Vermelha, Urca, Rio de Janeiro, R.J. (Brazil); Cruz, Leila Rosa de Oliveira [Military Institute of Engineering-IME, Pça. Gen. Tiburcio, 80, P. Vermelha, Urca, Rio de Janeiro, R.J. (Brazil); Campos, José Brant de [Rio de Janeiro State University - UERJ, Rio de Janeiro, R.J. (Brazil); Marçal, Rubens L. Santana Blazutti [Military Institute of Engineering-IME, Pça. Gen. Tiburcio, 80, P. Vermelha, Urca, Rio de Janeiro, R.J. (Brazil); Mijares, Dindo Q.; Coelho, Paulo G. [Department of Biomaterials and Biomimetics, New York University College of Dentistry (NYU), New York, NY (United States); Prado da Silva, Marcelo H. [Military Institute of Engineering-IME, Pça. Gen. Tiburcio, 80, P. Vermelha, Urca, Rio de Janeiro, R.J. (Brazil)

2017-01-01

In this work, two solutions were developed: the first, rich in Ca{sup 2+}, PO{sub 4}{sup 3−} ions and the second, rich in Ca{sup 2+}, PO{sub 4}{sup 3−} and Mg{sup 2+}, defined as Mg-modified precursor solution. For each Mg-modified precursor solution, the concentrations of Mg{sup 2+} ions were progressively increased by 5%, 10% and 15%wt. The aims of this research were to investigate the influence of magnesium ions substitution in calcium phosphate coatings on titanium surface and to evaluate these coatings by bioactivity assay in McCoy culture medium. The obtained coatings were characterized by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) analysis, and the presence of Mg ions was confirmed by the inductively coupled plasma atomic emission spectroscopy (ICP) analysis. In vitro bioactivity assay in McCoy culture medium showed bioactivity after 14 days in incubation for the HA and 10% Mg-monetite coatings. The high chemical stability of Mg-HA coatings was verified by the bioactivity assays, and no bone-like apatite deposition, characteristic of bioactivity, was observed for Mg-HA coatings, for the time period used in this study. - Highlights: • The presence of Mg ions influenced the final apatite phase present in the produced coatings. • A lower efficiency in heterogeneous deposition and an exposure of Ti substrate in 5% Mg-monetite coatings was soon verified. • McCoy culture medium was effective in predicting the coatings bioactivity.

Multilayer bioactive glass/zirconium titanate thin films in bone tissue engineering and regenerative dentistry

Science.gov (United States)

Mozafari, Masoud; Salahinejad, Erfan; Shabafrooz, Vahid; Yazdimamaghani, Mostafa; Vashaee, Daryoosh; Tayebi, Lobat

2013-01-01

Surface modification, particularly coatings deposition, is beneficial to tissue-engineering applications. In this work, bioactive glass/zirconium titanate composite thin films were prepared by a sol-gel spin-coating method. The surface features of the coatings were studied by scanning electron microscopy, atomic force microscopy, and spectroscopic reflection analyses. The results show that uniform and sound multilayer thin films were successfully prepared through the optimization of the process variables and the application of carboxymethyl cellulose as a dispersing agent. Also, it was found that the thickness and roughness of the multilayer coatings increase nonlinearly with increasing the number of the layers. This new class of nanocomposite coatings, comprising the bioactive and inert components, is expected not only to enhance bioactivity and biocompatibility, but also to protect the surface of metallic implants against wear and corrosion. PMID:23641155

Inhibition of Listeria monocytogenes ATCC 19115 on ham steak by tea bioactive compounds incorporated into chitosan-coated plastic films

Directory of Open Access Journals (Sweden)

Vodnar Dan C

2012-07-01

Full Text Available Abstract Background The consumer demands for better quality and safety of food products have given rise to the development and implementation of edible films. The use of antimicrobial films can be a promising tool for controlling L. monocytogenes on ready to eat products. The aim of this study was to develop effective antimicrobial films incorporating bioactive compounds from green and black teas into chitosan, for controlling L. monocytogenes ATCC 19115 on vacuum-packaged ham steak. The effectiveness of these antimicrobial films was evaluated at room temperature (20°C for 10 days and at refrigerated temperature (4°C for 8 weeks. Results The HPLC results clearly show that relative concentrations of catechins and caffeine in green tea ranked EGCG>EGC>CAF>ECG>EC>C while in black tea extracts ranked CAF>EGCG>ECG>EGC>EC>C. The chitosan-coated plastic films incorporating green tea and black tea extracts shows specific markers identified by FTIR. Incorporating natural extracts into chitosan showed that the growth of L monocytogenes ATCC 19115 was inhibited. The efficacy of antimicrobial effect of tea extracts incorporated into chitosan-coated plastic film was dose dependent. However, chitosan-coated films without addition of tea extracts did not inhibit the growth of L. monocytogenes ATCC 19115. Chitosan-coated plastic films incorporating 4% Green tea extract was the most effective antimicrobial, reducing the initial counts from 3.2 to 2.65 log CFU/cm2 during room temperature storage and from 3.2 to 1–1.5 log CFU/cm2 during refrigerated storage. Conclusions Incorporation of tea extracts into the chitosan-coated films considerably enhanced their effectiveness against L. monocytogenes ATCC 19115. 4% Green tea incorporated into chitosan-coated plastic film had a better antilisterial effect than 2% green tea or 2% and 4% black tea. Data from this study would provide new formulation options for developing antimicrobial packaging films using tea

Beta1 integrin-mediated adhesion signalling is essential for epidermal progenitor cell expansion.

Directory of Open Access Journals (Sweden)

Aleksandra Piwko-Czuchra

Full Text Available BACKGROUND: There is a major discrepancy between the in vitro and in vivo results regarding the role of beta1 integrins in the maintenance of epidermal stem/progenitor cells. Studies of mice with skin-specific ablation of beta1 integrins suggested that epidermis can form and be maintained in their absence, while in vitro data have shown a fundamental role for these adhesion receptors in stem/progenitor cell expansion and differentiation. METHODOLOGY/PRINCIPAL FINDINGS: To elucidate this discrepancy we generated hypomorphic mice expressing reduced beta1 integrin levels on keratinocytes that developed similar, but less severe defects than mice with beta1-deficient keratinocytes. Surprisingly we found that upon aging these abnormalities attenuated due to a rapid expansion of cells, which escaped or compensated for the down-regulation of beta1 integrin expression. A similar phenomenon was observed in aged mice with a complete, skin-specific ablation of the beta1 integrin gene, where cells that escaped Cre-mediated recombination repopulated the mutant skin in a very short time period. The expansion of beta1 integrin expressing keratinocytes was even further accelerated in situations of increased keratinocyte proliferation such as wound healing. CONCLUSIONS/SIGNIFICANCE: These data demonstrate that expression of beta1 integrins is critically important for the expansion of epidermal progenitor cells to maintain epidermal homeostasis.

The colocalization potential of HIV-specific CD8+ and CD4+ T-cells is mediated by integrin β7 but not CCR6 and regulated by retinoic acid.

Directory of Open Access Journals (Sweden)

Vanessa Sue Wacleche

Full Text Available CD4(+ T-cells from gut-associated lymphoid tissues (GALT are major targets for HIV-1 infection. Recruitment of excess effector CD8(+ T-cells in the proximity of target cells is critical for the control of viral replication. Here, we investigated the colocalization potential of HIV-specific CD8(+ and CD4(+ T-cells into the GALT and explored the role of retinoic acid (RA in regulating this process in a cohort of HIV-infected subjects with slow disease progression. The expression of the gut-homing molecules integrin β7, CCR6, and CXCR3 was identified as a "signature" for HIV-specific but not CMV-specific CD4(+ T-cells thus providing a new explanation for their enhanced permissiveness to infection in vivo. HIV-specific CD8(+ T-cells also expressed high levels of integrin β7 and CXCR3; however CCR6 was detected at superior levels on HIV-specific CD4(+ versus CD8(+ T-cells. All trans RA (ATRA upregulated the expression of integrin β7 but not CCR6 on HIV-specific T-cells. Together, these results suggest that HIV-specific CD8(+ T-cells may colocalize in excess with CD4(+ T-cells into the GALT via integrin β7 and CXCR3, but not via CCR6. Considering our previous findings that CCR6(+CD4(+ T-cells are major cellular targets for HIV-DNA integration in vivo, a limited ability of CD8(+ T-cells to migrate in the vicinity of CCR6(+CD4(+ T-cells may facilitate HIV replication and dissemination at mucosal sites.

Mechanical control of cyclic AMP signalling and gene transcription through integrins

Science.gov (United States)

Meyer, C. J.; Alenghat, F. J.; Rim, P.; Fong, J. H.; Fabry, B.; Ingber, D. E.

2000-01-01

This study was carried out to discriminate between two alternative hypotheses as to how cells sense mechanical forces and transduce them into changes in gene transcription. Do cells sense mechanical signals through generalized membrane distortion or through specific transmembrane receptors, such as integrins? Here we show that mechanical stresses applied to the cell surface alter the cyclic AMP signalling cascade and downstream gene transcription by modulating local release of signals generated by activated integrin receptors in a G-protein-dependent manner, whereas distortion of integrins in the absence of receptor occupancy has no effect.

Graphene-containing PCL- coated Porous 13-93B3 Bioactive Glass Scaffolds for Bone Regeneration

Science.gov (United States)

Türk, Mert; Deliormanlı, Aylin M.

2018-04-01

Borate-based 13-93B3 bioactive glass scaffolds were coated with the graphene-containing poly-caprolactone (PCL) solution to prepare electrically conductive composites for biomedical applications. Results revealed that electrical conductivity of the scaffolds increased with increasing concentration of graphene nanoparticles. Significant difference was not observed in hydroxyapatite forming ability of the bare and the graphene-containing scaffolds immersed in simulated body fluid. In vitro cytotoxicity experiments (XTT tests) showed that pre-osteoblastic MC3T3-E1 cell viability percentages of the graphene- containing samples was higher than control group samples after 7 days of incubation. However, a decrease in cell viability rates was obtained after 14 days of incubation for samples coated with PCL containing graphene starting from 3 wt%. Additionally, results obtained in the live-dead assay were consistent with the results of XTT tests. A higher ALP activity was detected in cells cultured on the graphene-containing borate glass scaffolds than those on the bare PCL coated 13-93B3 scaffolds suggesting the presence of graphene nanopowders stimulated an early stage of osteoblastic differentiation. SEM analysis showed that MC3T3-E1 cells exhibited a flat appearance and spread out through the surface in all groups of scaffolds starting from 3 days of incubation.

Leukocyte integrins and their ligand interactions

Science.gov (United States)

Hyun, Young-Min; Lefort, Craig T.; Kim, Minsoo

2010-01-01

Although critical for cell adhesion and migration during normal immune-mediated reactions, leukocyte integrins are also involved in the pathogenesis of diverse clinical conditions including autoimmune diseases and chronic inflammation. Leukocyte integrins therefore have been targets for anti-adhesive therapies to treat the inflammatory disorders. Recently, the therapeutic potential of integrin antagonists has been demonstrated in psoriasis and multiple sclerosis. However, current therapeutics broadly affect integrin functions and, thus, yield unfavorable side effects. This review discusses the major leukocyte integrins and the anti-adhesion strategies for treating immune diseases. PMID:19184539

Bioactive materials for biomedical applications using sol-gel technology

International Nuclear Information System (INIS)

Gupta, Radha; Kumar, Ashok

2008-01-01

This review paper focuses on the sol-gel technology that has been applied in many of the potential research areas and highlights the importance of sol-gel technology for preparing bioactive materials for biomedical applications. The versatility of sol-gel chemistry enables us to manipulate the characteristics of material required for particular applications. Sol-gel derived materials have proved to be good biomaterials for coating films and for the construction of super-paramagnetic nanoparticles, bioactive glasses and fiberoptic applicators for various biomedical applications. The introduction of the sol-gel route in a conventional method of preparing implants improves the mechanical strength, biocompatibility and bioactivity of scaffolds and prevents corrosion of metallic implants. The use of organically modified silanes (ORMOSILS) yields flexible and bioactive materials for soft and hard tissue replacement. A novel approach of nitric-oxide-releasing sol-gels as antibacterial coatings for reducing the infection around orthopedic implants has also been discussed

Mechanochemically synthesized kalsilite based bioactive glass-ceramic composite for dental vaneering

Science.gov (United States)

Kumar, Pattem Hemanth; Singh, Vinay Kumar; Kumar, Pradeep

2017-08-01

Kalsilite glass-ceramic composites have been prepared by a mechanochemical synthesis process for dental veneering application. The aim of the present study is to prepare bioactive kalsilite composite material for application in tissue attachment and sealing of the marginal gap between fixed prosthesis and tooth. Mechanochemical synthesis is used for the preparation of microfine kalsilite glass-ceramic. Low temperature frit and bioglass have been prepared using the traditional quench method. Thermal, microstructural and bioactive properties of the composite material have been examined. The feasibility of the kalsilite to be coated on the base commercial opaque as well as the bioactive behavior of the coated specimen has been confirmed. This study indicates that the prepared kalsilite-based composites show similar structural, morphological and bioactive behavior to that of commercial VITA VMK95 Dentin 1M2.

The effect of conditional inactivation of beta 1 integrins using twist 2 Cre, Osterix Cre and osteocalcin Cre lines on skeletal phenotype.

Science.gov (United States)

Shekaran, Asha; Shoemaker, James T; Kavanaugh, Taylor E; Lin, Angela S; LaPlaca, Michelle C; Fan, Yuhong; Guldberg, Robert E; García, Andrés J

2014-11-01

Skeletal development and growth are complex processes regulated by multiple microenvironmental cues, including integrin-ECM interactions. The β1 sub-family of integrins is the largest integrin sub-family and constitutes the main integrin binding partners of collagen I, the major ECM component of bone. As complete β1 integrin knockout results in embryonic lethality, studies of β1 integrin function in vivo rely on tissue-specific gene deletions. While multiple in vitro studies indicate that β1 integrins are crucial regulators of osteogenesis and mineralization, in vivo osteoblast-specific perturbations of β1 integrins have resulted in mild and sometimes contradictory skeletal phenotypes. To further investigate the role of β1 integrins on skeletal phenotype, we used the Twist2-Cre, Osterix-Cre and osteocalcin-Cre lines to generate conditional β1 integrin deletions, where Cre is expressed primarily in mesenchymal condensation, pre-osteoblast, and mature osteoblast lineage cells respectively within these lines. Mice with Twist2-specific β1 integrin disruption were smaller, had impaired skeletal development, especially in the craniofacial and vertebral tissues at E19.5, and did not survive beyond birth. Osterix-specific β1 integrin deficiency resulted in viable mice which were normal at birth but displayed early defects in calvarial ossification, incisor eruption and growth as well as femoral bone mineral density, structure, and mechanical properties. Although these defects persisted into adulthood, they became milder with age. Finally, a lack of β1 integrins in mature osteoblasts and osteocytes resulted in minor alterations to femur structure but had no effect on mineral density, biomechanics or fracture healing. Taken together, our data indicate that β1 integrin expression in early mesenchymal condensations play an important role in skeletal ossification, while β1 integrin-ECM interactions in pre-osteoblast, odontoblast- and hypertrophic chondryocyte

Medium-chain, triglyceride-containing lipid emulsions increase human neutrophil beta2 integrin expression, adhesion, and degranulation.

Science.gov (United States)

Wanten, G J; Geijtenbeek, T B; Raymakers, R A; van Kooyk, Y; Roos, D; Jansen, J B; Naber, A H

2000-01-01

To test the hypothesis that lipid emulsions with different triglyceride structures have distinct immunomodulatory properties, we analyzed human neutrophil adhesion and degranulation after lipid incubation. Neutrophils, isolated from the blood of 10 healthy volunteers, were incubated in medium or physiologic (2.5 mmol/L) emulsions containing long-chain (LCT), medium-chain (MCT), mixed LCT/MCT, or structured (SL) triglycerides. Expression of adhesion molecules and degranulation markers was evaluated by flow cytometry. Also, functional adhesion was investigated by means of a flow cytometric assay using fluorescent beads coated with the integrin ligand intercellular adhesion molecule (ICAM)-1. Although LCT and SL had no effect, LCT/MCT significantly increased expression of the beta2 integrins lymphocyte-function-associated antigen 1 (+18%), macrophage antigen 1 (+387%), p150,95 (+82%), and (alphaDbeta2 (+230%). Degranulation marker expression for azurophilic (CD63, +210%) and specific granules (CD66b, +370%) also significantly increased, whereas L-selectin (CD62L, -70%) decreased. The effects of LCT/MCT were mimicked by the MCT emulsion. ICAM-1 adhesion (% beads bound) was increased by LCT/MCT (34% +/- 4%), whereas LCT (19% +/-3%) and SL (20% +/- 2%) had no effect compared with medium (17% +/- 3%). LCT/MCT and MCT, contrary to LCT and SL emulsions, increased neutrophil beta2 integrin expression, adhesion, and degranulation. Apart from other emulsion constituents, triglyceride chain length might therefore be a key feature in the interaction of lipid emulsions and the phagocyte immune system.

Applications of snake venom components to modulate integrin activities in cell-matrix interactions

Science.gov (United States)

Marcinkiewicz, Cezary

2013-01-01

Snake venom proteins are broadly investigated in the different areas of life science. Direct interaction of these compounds with cells may involve a variety of mechanisms that result in diverse cellular responses leading to the activation or blocking of physiological functions of the cell. In this review, the snake venom components interacting with integrins will be characterized in context of their effect on cellular response. Currently, two major families of snake venom proteins are considered as integrin-binding molecules. The most attention has been devoted to the disintegrin family, which binds certain types of integrins through specific motifs recognized as a tri-peptide structurally localized on an integrin-binding loop. Other snake venom integrin-binding proteins belong to the C-type lectin family. Snake venom molecules bind to the cellular integrins resulting in a modulation of cell signaling and in consequence, the regulation of cell proliferation, migration and apoptosis. Therefore, snake venom research on the integrin-binding molecules may have significance in biomedicine and basic cell biology. PMID:23811033

Integrins and extracellular matrix in mechanotransduction

Directory of Open Access Journals (Sweden)

Ramage L

2011-12-01

Full Text Available Lindsay RamageQueen’s Medical Research Institute, University of Edinburgh, Edinburgh, UKAbstract: Integrins are a family of cell surface receptors which mediate cell–matrix and cell–cell adhesions. Among other functions they provide an important mechanical link between the cells external and intracellular environments while the adhesions that they form also have critical roles in cellular signal-transduction. Cell–matrix contacts occur at zones in the cell surface where adhesion receptors cluster and when activated the receptors bind to ligands in the extracellular matrix. The extracellular matrix surrounds the cells of tissues and forms the structural support of tissue which is particularly important in connective tissues. Cells attach to the extracellular matrix through specific cell-surface receptors and molecules including integrins and transmembrane proteoglycans. Integrins work alongside other proteins such as cadherins, immunoglobulin superfamily cell adhesion molecules, selectins, and syndecans to mediate cell–cell and cell–matrix interactions and communication. Activation of adhesion receptors triggers the formation of matrix contacts in which bound matrix components, adhesion receptors, and associated intracellular cytoskeletal and signaling molecules form large functional, localized multiprotein complexes. Cell–matrix contacts are important in a variety of different cell and tissue properties including embryonic development, inflammatory responses, wound healing, and adult tissue homeostasis. This review summarizes the roles and functions of integrins and extracellular matrix proteins in mechanotransduction.Keywords: ligand binding, α subunit, ß subunit, focal adhesion, cell differentiation, mechanical loading, cell–matrix interaction

Integrin Signalling

OpenAIRE

Schelfaut, Roselien

2005-01-01

Integrins are receptors presented on most cells. By binding ligand they can generate signalling pathways inside the cell. Those pathways are a linkage to proteins in the cytosol. It is known that tumor cells can survive and proliferate in the absence of a solid support while normal cells need to be bound to ligand. To understand why tumour cells act that way, we first have to know how ligand-binding to integrins affect the cell. This research field includes studies on activation of proteins b...

Sequence-specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98.

Science.gov (United States)

Takahashi, Y; Bigler, D; Ito, Y; White, J M

2001-04-01

ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion. Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs. Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain. Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop. Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface. Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop. Bead binding was also inhibited by the function-blocking anti-alpha6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-alpha6 mAb, or by mAbs against either the alphav or beta3 integrin subunits. We also present evidence that in addition to the tetraspanin CD9, two other beta1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs. Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain. Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg beta1 integrin-associated proteins in sperm-egg binding and fusion. We propose that an egg surface "tetraspan web" facilitates fertilization and that it may do so by fostering ADAM-integrin interactions.

Sequence-Specific Interaction between the Disintegrin Domain of Mouse ADAM 3 and Murine Eggs: Role of β1 Integrin-associated Proteins CD9, CD81, and CD98

Science.gov (United States)

Takahashi, Yuji; Bigler, Dora; Ito, Yasuhiko; White, Judith M.

2001-01-01

ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion. Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs. Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain. Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop. Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface. Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop. Bead binding was also inhibited by the function-blocking anti-α6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-α6 mAb, or by mAbs against either the αv or β3 integrin subunits. We also present evidence that in addition to the tetraspanin CD9, two other β1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs. Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain. Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg β1 integrin-associated proteins in sperm-egg binding and fusion. We propose that an egg surface “tetraspan web” facilitates fertilization and that it may do so by fostering ADAM–integrin interactions. PMID:11294888

Bioactive glass-ceramic coatings prepared by pulsed laser deposition from RKKP targets (sol-gel vs melt-processing route)

Energy Technology Data Exchange (ETDEWEB)

Rau, J.V., E-mail: giulietta.rau@ism.cnr.it [Istituto di Struttura della Materia, Consiglio Nazionale delle Ricerche, Via del Fosso del Cavaliere, 100-00133 Rome (Italy); Teghil, R. [Universita della Basilicata, Dipartimento di Chimica ' A.M. Tamburro' , Via dell' Ateneo Lucano, 10-85100 Potenza (Italy); CNR-IMIP U.O.S. di Potenza, Zona Industriale di Tito scalo (PZ) (Italy); Fosca, M. [Istituto di Struttura della Materia, Consiglio Nazionale delle Ricerche, Via del Fosso del Cavaliere, 100-00133 Rome (Italy); Universita di Roma ' La Sapienza' , Dipartimento di Chimica, Piazzale Aldo Moro, 5-00185 Rome (Italy); De Bonis, A. [Universita della Basilicata, Dipartimento di Chimica ' A.M. Tamburro' , Via dell' Ateneo Lucano, 10-85100 Potenza (Italy); CNR-IMIP U.O.S. di Potenza, Zona Industriale di Tito scalo (PZ) (Italy); Cacciotti, I.; Bianco, A. [Universita di Roma ' Tor Vergata' , Dipartimento di Ingegneria Industriale, UR INSTM ' Roma Tor Vergata' , Via del Politecnico, 1-00133 Rome (Italy); Albertini, V. Rossi [Istituto di Struttura della Materia, Consiglio Nazionale delle Ricerche, Via del Fosso del Cavaliere, 100-00133 Rome (Italy); Caminiti, R. [Universita di Roma ' La Sapienza' , Dipartimento di Chimica, Piazzale Aldo Moro, 5-00185 Rome (Italy); Ravaglioli, A. [Parco Torricelli delle Arti e delle Scienze, Via Granarolo, 64-48018 Faenza (Ra) (Italy)

2012-05-15

Highlights: Black-Right-Pointing-Pointer Bioactive glass-ceramic coatings for bone tissue repair and regeneration. Black-Right-Pointing-Pointer Pulsed Lased Deposition allowed congruent transfer of target composition to coating. Black-Right-Pointing-Pointer Target was prepared by sol-gel process suitable for compositional tailoring. Black-Right-Pointing-Pointer Titanium, widely used for orthopaedics and dental implants, was used as substrate. Black-Right-Pointing-Pointer The physico-chemical properties of the prepared coatings are reported. -- Abstract: The deposition of innovative glass-ceramic composition (i.e. RKKP) coatings by Pulsed Lased Deposition (PLD) technique is reported. RKKP was synthesised following two methodologies: melt-processing and sol-gel, the latter being particularly suitable to tailor the compositional range. The PLD advantage with respect to other deposition techniques is the congruent transfer of the target composition to the coating. The physico-chemical properties of films were investigated by Scanning Electron and Atomic Force Microscopies, Fourier Transform Infrared Spectroscopy, Angular and Energy Dispersive X-ray Diffraction, and Vickers microhardness. The deposition performed at 12 J/cm{sup 2} and 500 Degree-Sign C allows to prepare crystalline films with the composition that replicates rather well that of the initial targets. The 0.6 {mu}m thin melt-processing RKKP films, possessing the hardness of 25 GPa, and the 4.3 {mu}m thick sol-gel films with the hardness of 17 GPa were obtained.

Bioactive glass–ceramic coatings prepared by pulsed laser deposition from RKKP targets (sol–gel vs melt-processing route)

International Nuclear Information System (INIS)

Rau, J.V.; Teghil, R.; Fosca, M.; De Bonis, A.; Cacciotti, I.; Bianco, A.; Albertini, V. Rossi; Caminiti, R.; Ravaglioli, A.

2012-01-01

Highlights: ► Bioactive glass–ceramic coatings for bone tissue repair and regeneration. ► Pulsed Lased Deposition allowed congruent transfer of target composition to coating. ► Target was prepared by sol–gel process suitable for compositional tailoring. ► Titanium, widely used for orthopaedics and dental implants, was used as substrate. ► The physico-chemical properties of the prepared coatings are reported. -- Abstract: The deposition of innovative glass–ceramic composition (i.e. RKKP) coatings by Pulsed Lased Deposition (PLD) technique is reported. RKKP was synthesised following two methodologies: melt-processing and sol–gel, the latter being particularly suitable to tailor the compositional range. The PLD advantage with respect to other deposition techniques is the congruent transfer of the target composition to the coating. The physico-chemical properties of films were investigated by Scanning Electron and Atomic Force Microscopies, Fourier Transform Infrared Spectroscopy, Angular and Energy Dispersive X-ray Diffraction, and Vickers microhardness. The deposition performed at 12 J/cm 2 and 500 °C allows to prepare crystalline films with the composition that replicates rather well that of the initial targets. The 0.6 μm thin melt-processing RKKP films, possessing the hardness of 25 GPa, and the 4.3 μm thick sol–gel films with the hardness of 17 GPa were obtained.

Effect of CeO2 and Y2O3 on microstructure, bioactivity and degradability of laser cladding CaO-SiO2 coating on titanium alloy.

Science.gov (United States)

Li, H C; Wang, D G; Chen, C Z; Weng, F

2015-03-01

To solve the lack of strength of bulk biomaterials for load-bearing applications and improve the bioactivity of titanium alloy (Ti-6Al-4V), CaO-SiO2 coatings on titanium alloy were fabricated by laser cladding technique. The effect of CeO2 and Y2O3 on microstructure and properties of laser cladding coating was analyzed. The cross-section microstructure of ceramic layer from top to bottom gradually changes from cellular-dendrite structure to compact cellular crystal. The addition of CeO2 or Y2O3 refines the microstructure of the ceramic layer in the upper and middle regions. The refining effect on the grain is related to the kinds of additives and their content. The coating is mainly composed of CaTiO3, CaO, α-Ca2(SiO4), SiO2 and TiO2. Y2O3 inhibits the formation of CaO. After soaking in simulated body fluid (SBF), the calcium phosphate layer is formed on the coating surface, indicating the coating has bioactivity. After soaking in Tris-HCl solution, the samples doped with CeO2 or Y2O3 present a lower weight loss, indicating the addition of CeO2 or Y2O3 improves the degradability of laser cladding sample. Copyright © 2015 Elsevier B.V. All rights reserved.

Long-term liver-specific functions of hepatocytes in electrospun chitosan nanofiber scaffolds coated with fibronectin.

Science.gov (United States)

Rajendran, Divya; Hussain, Ali; Yip, Derek; Parekh, Amit; Shrirao, Anil; Cho, Cheul H

2017-08-01

In this study, a new 3D liver model was developed using biomimetic nanofiber scaffolds and co-culture system consisting of hepatocytes and fibroblasts for the maintenance of long-term liver functions. The chitosan nanofiber scaffolds were fabricated by the electrospinning technique. To enhance cellular adhesion and spreading, the surfaces of the chitosan scaffolds were coated with fibronectin (FN) by adsorption and evaluated for various cell types. Cellular phenotype, protein expression, and liver-specific functions were extensively characterized by immunofluorescent and histochemical stainings, albumin enzyme-linked immunosorbent assay and Cytochrome p450 detoxification assays, and scanning electron microscopy. The electrospun chitosan scaffolds exhibited a highly porous and randomly oriented nanofibrous structure. The FN coating on the surface of the chitosan nanofibers significantly enhanced cell attachment and spreading, as expected, as surface modification with this cell adhesion molecule on the chitosan surface is important for focal adhesion formation and integrin binding. Comparison of hepatocyte mono-cultures and co-cultures in 3D culture systems indicated that the hepatocytes in co-cultures formed colonies and maintained their morphologies and functions for prolonged periods of time. The 3D liver tissue model developed in this study will provide useful tools toward the development of engineered liver tissues for drug screening and tissue engineering applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2119-2128, 2017. © 2017 Wiley Periodicals, Inc.

Bioactive lipid coating of bone allografts directs engraftment and fate determination of bone marrow-derived cells in rat GFP chimeras.

Science.gov (United States)

Das, Anusuya; Segar, Claire E; Chu, Yihsuan; Wang, Tiffany W; Lin, Yong; Yang, Chunxi; Du, Xeujun; Ogle, Roy C; Cui, Quanjun; Botchwey, Edward A

2015-09-01

Bone grafting procedures are performed to treat wounds incurred during wartime trauma, accidents, and tumor resections. Endogenous mechanisms of repair are often insufficient to ensure integration between host and donor bone and subsequent restoration of function. We investigated the role that bone marrow-derived cells play in bone regeneration and sought to increase their contributions by functionalizing bone allografts with bioactive lipid coatings. Polymer-coated allografts were used to locally deliver the immunomodulatory small molecule FTY720 in tibial defects created in rat bone marrow chimeras containing genetically-labeled bone marrow for monitoring cell origin and fate. Donor bone marrow contributed significantly to both myeloid and osteogenic cells in remodeling tissue surrounding allografts. FTY720 coatings altered the phenotype of immune cells two weeks post-injury, which was associated with increased vascularization and bone formation surrounding allografts. Consequently, degradable polymer coating strategies that deliver small molecule growth factors such as FTY720 represent a novel therapeutic strategy for harnessing endogenous bone marrow-derived progenitors and enhancing healing in load-bearing bone defects. Copyright © 2015 Elsevier Ltd. All rights reserved.

The Integrin Receptor in Biologically Relevant Bilayers

DEFF Research Database (Denmark)

Kalli, Antreas C.; Róg, Tomasz; Vattulainen, Ilpo

2017-01-01

/talin complex was inserted in biologically relevant bilayers that resemble the cell plasma membrane containing zwitterionic and charged phospholipids, cholesterol and sphingolipids to study the dynamics of the integrin receptor and its effect on bilayer structure and dynamics. The results of this study...... demonstrate the dynamic nature of the integrin receptor and suggest that the presence of the integrin receptor alters the lipid organization between the two leaflets of the bilayer. In particular, our results suggest elevated density of cholesterol and of phosphatidylserine lipids around the integrin....../talin complex and a slowing down of lipids in an annulus of ~30 Å around the protein due to interactions between the lipids and the integrin/talin F2–F3 complex. This may in part regulate the interactions of integrins with other related proteins or integrin clustering thus facilitating signal transduction...

Tensin stabilizes integrin adhesive contacts in Drosophila.

Science.gov (United States)

Torgler, Catherine N; Narasimha, Maithreyi; Knox, Andrea L; Zervas, Christos G; Vernon, Matthew C; Brown, Nicholas H

2004-03-01

We report the functional characterization of the Drosophila ortholog of tensin, a protein implicated in linking integrins to the cytoskeleton and signaling pathways. A tensin null was generated and is viable with wing blisters, a phenotype characteristic of loss of integrin adhesion. In tensin mutants, mechanical abrasion is required during wing expansion to cause wing blisters, suggesting that tensin strengthens integrin adhesion. The localization of tensin requires integrins, talin, and integrin-linked kinase. The N-terminal domain and C-terminal PTB domain of tensin provide essential recruitment signals. The intervening SH2 domain is not localized on its own. We suggest a model where tensin is recruited to sites of integrin adhesion via its PTB and N-terminal domains, localizing the SH2 domain so that it can interact with phosphotyrosine-containing proteins, which stabilize the integrin link to the cytoskeleton.

Ca2+-dependent localization of integrin-linked kinase to cell junctions in differentiating keratinocytes.

Science.gov (United States)

Vespa, Alisa; Darmon, Alison J; Turner, Christopher E; D'Souza, Sudhir J A; Dagnino, Lina

2003-03-28

Integrin complexes are necessary for proper proliferation and differentiation of epidermal keratinocytes. Differentiation of these cells is accompanied by down-regulation of integrins and focal adhesions as well as formation of intercellular adherens junctions through E-cadherin homodimerization. A central component of integrin adhesion complexes is integrin-linked kinase (ILK), which can induce loss of E-cadherin expression and epithelial-mesenchymal transformation when ectopically expressed in intestinal and mammary epithelia. In cultured primary mouse keratinocytes, we find that ILK protein levels are independent of integrin expression and signaling, since they remain constant during Ca(2+)-induced differentiation. In contrast, keratinocyte differentiation is accompanied by marked reduction in kinase activity in ILK immunoprecipitates and altered ILK subcellular distribution. Specifically, ILK distributes in close apposition to actin fibers along intercellular junctions in differentiated but not in undifferentiated keratinocytes. ILK localization to cell-cell borders occurs independently of integrin signaling and requires Ca(2+) as well as an intact actin cytoskeleton. Further, and in contrast to what is observed in other epithelial cells, ILK overexpression in differentiated keratinocytes does not promote E-cadherin down-regulation and epithelial-mesenchymal transition. Thus, novel tissue-specific mechanisms control the formation of ILK complexes associated with cell-cell junctions in differentiating murine epidermal keratinocytes.

Sputtered titanium oxynitride coatings for endosseous applications: Physical and chemical evaluation and first bioactivity assays

Energy Technology Data Exchange (ETDEWEB)

Banakh, Oksana, E-mail: oksana.banakh@he-arc.ch [Institute of Applied Microtechnologies, Haute Ecole Arc Ingénierie (HES-SO), Eplatures-Grise 17, CH-2300 La Chaux-de-Fonds (Switzerland); Moussa, Mira, E-mail: mira.moussa@unige.ch [Laboratory of Biomaterials, University of Geneva, 19, rue Barthelemy Menn, CH-1205 Geneva (Switzerland); Matthey, Joel, E-mail: joel.matthey@he-arc.ch [Institute of Applied Microtechnologies, Haute Ecole Arc Ingénierie (HES-SO), Eplatures-Grise 17, CH-2300 La Chaux-de-Fonds (Switzerland); Pontearso, Alessandro, E-mail: alessandro.pontearso@he-arc.ch [Institute of Applied Microtechnologies, Haute Ecole Arc Ingénierie (HES-SO), Eplatures-Grise 17, CH-2300 La Chaux-de-Fonds (Switzerland); Cattani-Lorente, Maria, E-mail: maria.cattani-lorente@unige.ch [Laboratory of Biomaterials, University of Geneva, 19, rue Barthelemy Menn, CH-1205 Geneva (Switzerland); Sanjines, Rosendo, E-mail: rosendo.sanjines@epfl.ch [Ecole Polytechnique Fédérale de Lausanne (EPFL), Institute of Condensed Matter Physics, Station 3, CH-1015 Lausanne (Switzerland); Fontana, Pierre, E-mail: Pierre.Fontana@hcuge.ch [Haemostasis laboratory, Geneva University Hospital, Rue Gabrielle-Perret-Gentil 4, CH-1205 Geneva (Switzerland); Wiskott, Anselm, E-mail: anselm.wiskott@unige.ch [Laboratory of Biomaterials, University of Geneva, 19, rue Barthelemy Menn, CH-1205 Geneva (Switzerland); Durual, Stephane, E-mail: stephane.durual@unige.ch [Laboratory of Biomaterials, University of Geneva, 19, rue Barthelemy Menn, CH-1205 Geneva (Switzerland)

2014-10-30

Highlights: • Titanium oxynitride coatings (TiN{sub x}O{sub y}) with chemical composition ranging from TiN to TiO{sub 2} were deposited by magnetron sputtering from a metallic Ti target using a mixture of O{sub 2} + N{sub 2}. • The coatings structure as well as physical, chemical and mechanical properties progressively changes as a function of oxygen content in the TiN{sub x}O{sub y.} • All TiN{sub x}O{sub y} coatings show a significantly higher level of bioactivity as compared to bare Ti substrates (1.2 to 1.4 fold increase in cell proliferation). Despite variations in surface chemistry, topography and surface tension observed on films as a function of chemical composition, no significant differences in the films’ biological activity were observed after 3 days of testing. - Abstract: Titanium oxynitride coatings (TiN{sub x}O{sub y}) are considered a promising material for applications in dental implantology due to their high corrosion resistance, their biocompatibility and their superior hardness. Using the sputtering technique, TiN{sub x}O{sub y} films with variable chemical compositions can be deposited. These films may then be set to a desired value by varying the process parameters, that is, the oxygen and nitrogen gas flows. To improve the control of the sputtering process with two reactive gases and to achieve a variable and controllable coating composition, the plasma characteristics were monitored in-situ by optical emission spectroscopy. TiN{sub x}O{sub y} films were deposited onto commercially pure (ASTM 67) microroughened titanium plates by reactive magnetron sputtering. The nitrogen gas flow was kept constant while the oxygen gas flow was adjusted for each deposition run to obtain films with different oxygen and nitrogen contents. The physical and chemical properties of the deposited films were analyzed as a function of oxygen content in the titanium oxynitride. The potential application of the coatings in dental implantology was assessed by

Porous biomorphic silicon carbide ceramics coated with hydroxyapatite as prospective materials for bone implants

Energy Technology Data Exchange (ETDEWEB)

Gryshkov, Oleksandr, E-mail: gryshkov@imp.uni-hannover.de [Institute for Multiphase Processes, Leibniz Universität Hannover, 30167 Hannover (Germany); Klyui, Nickolai I., E-mail: klyuini@ukr.net [College of Physics, Jilin University, 130012 Changchun (China); V. Lashkaryov Institute of Semiconductor Physics, National Academy of Science of Ukraine, 03028 Kyiv (Ukraine); Temchenko, Volodymyr P., E-mail: tvp@isp.kiev.ua [V. Lashkaryov Institute of Semiconductor Physics, National Academy of Science of Ukraine, 03028 Kyiv (Ukraine); Kyselov, Vitalii S., E-mail: kyselov@isp.kiev.ua [V. Lashkaryov Institute of Semiconductor Physics, National Academy of Science of Ukraine, 03028 Kyiv (Ukraine); Chatterjee, Anamika, E-mail: chatterjee@imp.uni-hannover.de [Institute for Multiphase Processes, Leibniz Universität Hannover, 30167 Hannover (Germany); Belyaev, Alexander E., E-mail: belyaev@isp.kiev.ua [V. Lashkaryov Institute of Semiconductor Physics, National Academy of Science of Ukraine, 03028 Kyiv (Ukraine); Lauterboeck, Lothar, E-mail: lauterboeck@imp.uni-hannover.de [Institute for Multiphase Processes, Leibniz Universität Hannover, 30167 Hannover (Germany); Iarmolenko, Dmytro, E-mail: iarmolenko.dmytro@isp.kiev.ua [V. Lashkaryov Institute of Semiconductor Physics, National Academy of Science of Ukraine, 03028 Kyiv (Ukraine); Glasmacher, Birgit, E-mail: glasmacher@imp.uni-hannover.de [Institute for Multiphase Processes, Leibniz Universität Hannover, 30167 Hannover (Germany)

2016-11-01

Porous and cytocompatible silicon carbide (SiC) ceramics derived from wood precursors and coated with bioactive hydroxyapatite (HA) and HA-zirconium dioxide (HA/ZrO{sub 2}) composite are materials with promising application in engineering of bone implants due to their excellent mechanical and structural properties. Biomorphic SiC ceramics have been synthesized from wood (Hornbeam, Sapele, Tilia and Pear) using a forced impregnation method. The SiC ceramics have been coated with bioactive HA and HA/ZrO{sub 2} using effective gas detonation deposition approach (GDD). The surface morphology and cytotoxicity of SiC ceramics as well as phase composition and crystallinity of deposited coatings were analyzed. It has been shown that the porosity and pore size of SiC ceramics depend on initial wood source. The XRD and FTIR studies revealed the preservation of crystal structure and phase composition of in the HA coating, while addition of ZrO{sub 2} to the initial HA powder resulted in significant decomposition of the final HA/ZrO{sub 2} coating and formation of other calcium phosphate phases. In turn, NIH 3T3 cells cultured in medium exposed to coated and uncoated SiC ceramics showed high re-cultivation efficiency as well as metabolic activity. The recultivation efficiency of cells was the highest for HA-coated ceramics, whereas HA/ZrO{sub 2} coating improved the recultivation efficiency of cells as compared to uncoated SiC ceramics. The GDD method allowed generating homogeneous HA coatings with no change in calcium to phosphorus ratio. In summary, porous and cytocompatible bio-SiC ceramics with bioactive coatings show a great promise in construction of light, robust, inexpensive and patient-specific bone implants for clinical application. - Highlights: • Synthesis and characterization of porous biomorphic SiC ceramics derived from wood • Successful deposition of bioactive calcium phosphate coatings using gas detonation deposition • Porosity and pore size of Si

Porous biomorphic silicon carbide ceramics coated with hydroxyapatite as prospective materials for bone implants

International Nuclear Information System (INIS)

Gryshkov, Oleksandr; Klyui, Nickolai I.; Temchenko, Volodymyr P.; Kyselov, Vitalii S.; Chatterjee, Anamika; Belyaev, Alexander E.; Lauterboeck, Lothar; Iarmolenko, Dmytro; Glasmacher, Birgit

2016-01-01

Porous and cytocompatible silicon carbide (SiC) ceramics derived from wood precursors and coated with bioactive hydroxyapatite (HA) and HA-zirconium dioxide (HA/ZrO 2 ) composite are materials with promising application in engineering of bone implants due to their excellent mechanical and structural properties. Biomorphic SiC ceramics have been synthesized from wood (Hornbeam, Sapele, Tilia and Pear) using a forced impregnation method. The SiC ceramics have been coated with bioactive HA and HA/ZrO 2 using effective gas detonation deposition approach (GDD). The surface morphology and cytotoxicity of SiC ceramics as well as phase composition and crystallinity of deposited coatings were analyzed. It has been shown that the porosity and pore size of SiC ceramics depend on initial wood source. The XRD and FTIR studies revealed the preservation of crystal structure and phase composition of in the HA coating, while addition of ZrO 2 to the initial HA powder resulted in significant decomposition of the final HA/ZrO 2 coating and formation of other calcium phosphate phases. In turn, NIH 3T3 cells cultured in medium exposed to coated and uncoated SiC ceramics showed high re-cultivation efficiency as well as metabolic activity. The recultivation efficiency of cells was the highest for HA-coated ceramics, whereas HA/ZrO 2 coating improved the recultivation efficiency of cells as compared to uncoated SiC ceramics. The GDD method allowed generating homogeneous HA coatings with no change in calcium to phosphorus ratio. In summary, porous and cytocompatible bio-SiC ceramics with bioactive coatings show a great promise in construction of light, robust, inexpensive and patient-specific bone implants for clinical application. - Highlights: • Synthesis and characterization of porous biomorphic SiC ceramics derived from wood • Successful deposition of bioactive calcium phosphate coatings using gas detonation deposition • Porosity and pore size of SiC ceramics depend on wood

Selective Modulation of Integrin-mediated Cell Migration by Distinct ADAM Family MembersV⃞

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Huang, Jing; Bridges, Lance C.; White, Judith M.

2005-01-01

A disintegrin and a metalloprotease (ADAM) family members have been implicated in many biological processes. Although it is recognized that recombinant ADAM disintegrin domains can interact with integrins, little is known about ADAM-integrin interactions in cellular context. Here, we tested whether ADAMs can selectively regulate integrin-mediated cell migration. ADAMs were expressed in Chinese hamster ovary cells that express defined integrins (α4β1, α5β1, or both), and cell migration on full-length fibronectin or on its α4β1 or α5β1 binding fragments was studied. We found that ADAMs inhibit integrin-mediated cell migration in patterns dictated by the integrin binding profiles of their isolated disintegrin domains. ADAM12 inhibited cell migration mediated by the α4β1 but not the α5β1 integrin. ADAM17 had the reciprocal effect; it inhibited α5β1- but not α4β1-mediated cell migration. ADAM19 and ADAM33 inhibited migration mediated by both α4β1 and α5β1 integrins. A point mutation in the ADAM12 disintegrin loop partially reduced the inhibitory effect of ADAM12 on cell migration on the α4β1 binding fragment of fibronectin, whereas mutations that block metalloprotease activity had no effect. Our results indicate that distinct ADAMs can modulate cell migration mediated by specific integrins in a pattern dictated, at least in part, by their disintegrin domains. PMID:16079176

Small molecule antagonists of integrin receptors.

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Perdih, A; Dolenc, M Sollner

2010-01-01

The complex and widespread family of integrin receptors is involved in numerous physiological processes, such as tissue remodeling, angiogenesis, development of the immune response and homeostasis. In addition, their key role has been elucidated in important pathological disorders such as cancer, cardiovascular diseases, osteoporosis, autoimmune and inflammatory diseases and in the pathogenesis of infectious diseases, making them highly important targets for modern drug design campaigns. In this review we seek to present a concise overview of the small molecule antagonists of this diverse and highly complex receptor family. Integrin antagonists are classified according to the targeted integrin receptor and are discussed in four sections. First we present the fibrinogen alpha(IIb)beta3 and the vitronectin alpha (V)beta(3) receptor antagonists. The remaining selective integrin antagonists are examined in the third section. The final section is dedicated to molecules with dual or multiple integrin activity. In addition, the use of antibodies and peptidomimetic approaches to modulate the integrin receptors are discussed, as well providing the reader with an overall appreciation of the field.

Effect of surface topography and bioactive properties on early adhesion and growth behavior of mouse preosteoblast MC3T3-E1 cells.

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Li, Na; Chen, Gang; Liu, Jue; Xia, Yang; Chen, Hanbang; Tang, Hui; Zhang, Feimin; Gu, Ning

2014-10-08

The effects of bioactive properties and surface topography of biomaterials on the adhesion and spreading properties of mouse preosteoblast MC3T3-E1 cells was investigated by preparation of different surfaces. Poly lactic-co-glycolic acid (PLGA) electrospun fibers (ES) were produced as a porous rough surface. In our study, coverslips were used as a substrate for the immobilization of 3,4-dihydroxyphenylalanine (DOPA) and collagen type I (COL I) in the preparation of bioactive surfaces. In addition, COL I was immobilized onto porous electrospun fibers surfaces (E-COL) to investigate the combined effects of bioactive molecules and topography. Untreated coverslips were used as controls. Early adhesion and growth behavior of MC3T3-E1 cells cultured on the different surfaces were studied at 6, 12, and 24 h. Evaluation of cell adhesion and morphological changes showed that the all the surfaces were favorable for promoting the adhesion and spreading of cells. CCK-8 assays and flow cytometry revealed that both topography and bioactive properties were favorable for cell growth. Analysis of β1, α1, α2, α5, α10 and α11 integrin expression levels by immunofluorescence, real-time RT-PCR, and Western blot and indicated that surface topography plays an important role in the early stage of cell adhesion. However, the influence of topography and bioactive properties of surfaces on integrins is variable. Compared with any of the topographic or bioactive properties in isolation, the combined effect of both types of properties provided an advantage for the growth and spreading of MC3T3-E1 cells. This study provides a new insight into the functions and effects of topographic and bioactive modifications of surfaces at the interface between cells and biomaterials for tissue engineering.

Long-term antibiotic delivery by chitosan-based composite coatings with bone regenerative potential

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Ordikhani, F.; Simchi, A.

2014-10-01

Composite coatings with bone-bioactivity and drug-eluting capacity are considered as promising materials for titanium bone implants. In this work, drug-eluting chitosan-bioactive glass coatings were fabricated by a single-step electrophoretic deposition technique. Drug-loading and -releasing capacity of the composite coatings were carried out using the vancomycin antibiotic. Uniform coatings with a thickness of ∼55 μm containing 23.7 wt% bioactive glass particles and various amounts of the antibiotic (380-630 μg/cm2) were produced. The coatings were bioactive in terms of apatite-forming ability in simulated body fluid and showed favorable cell adhesion and growth. In vitro biological tests also indicated that the composite coatings had better cellular affinity than pristine chitosan coatings. The in vitro elution kinetics of the composite coating revealed an initial burst release of around 40% of the drug within the first elution step of 1 h and following by a continuous eluting over 4 weeks, revealing long-term drug-delivering potential. Antibacterial tests using survival assay against Gram-positive Staphylococcus aureus bacteria determined the effect of vancomycin release on reduction of infection risk. Almost no bacteria were survived on the coatings prepared from the EPD suspension containing ≥0.5 g/l vancomycin. The developed chitosan-based composite coatings with bone bioactivity and long-term drug-delivery ability may be potentially useful for metallic implants to reduce infection risk.

Superior integrin activating capacity and higher adhesion to fibrinogen matrix in buffy coat-derived platelet concentrates (PCs) compared to PRP-PCs.

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Beshkar, Pezhman; Hosseini, Ehteramolsadat; Ghasemzadeh, Mehran

2018-02-01

Regardless of different sources, methods or devices which are applied for preparation of therapeutic platelets, these products are generally isolated from whole blood by the sedimentation techniques which are based on PRP or buffy coat (BC) separation. As a general fact, platelet preparation and storage are also associated with some deleterious changes that known as platelet storage lesion (PSL). Although these alternations in platelet functional activity are aggravated during storage, whether technical issues within preparation can affect integrin activation and platelet adhesion to fibrinogen were investigated in this study. PRP- and BC-platelet concentrates (PCs) were subjected to flowcytometry analysis to examine the expression of platelet activation marker, P-selectin as well as active confirmation of the GPIIb/IIIa (α IIb β 3 ) on day 0, 1, 3 and 5 post-storage. Platelet adhesion to fibrinogen matrix was evaluated by fluorescence microscopy. Glucose concentration and LDH activity were also measured by colorimetric methods. The increasing P-selectin expression during storage was in a reverse correlation with PAC-1 binding (r = -0.67; p = .001). PRP-PCs showed the higher level of P-selectin expression than BC-PCs, whereas the levels of PAC-1 binding and platelet adhesion to fibrinogen matrix were significantly lower in PRP-PCs. Higher levels of active confirmation of the GPIIb/IIIa in BC-PCs were also associated with greater concentration of glucose in these products. We demonstrated the superior capacities of integrin activation and adhesion to fibrinogen for BC-PCs compared to those of PRP-PCs. These findings may provide more advantages for BC method of platelet preparation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Force via integrins but not E-cadherin decreases Oct3/4 expression in embryonic stem cells

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Uda, Yuhei [Department of Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801 (United States); Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, 6-6-01, Aramaki-aoba, Aoba-ward, Sendai City (Japan); Poh, Yeh-Chuin; Chowdhury, Farhan; Wu, Douglas C. [Department of Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801 (United States); Tanaka, Tetsuya S. [Department of Animal Sciences, University of Illinois at Urbana-Champaign, Urbana, IL 61801 (United States); Sato, Masaaki [Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, 6-6-01, Aramaki-aoba, Aoba-ward, Sendai City (Japan); Wang, Ning, E-mail: nwangrw@illinois.edu [Department of Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801 (United States)

2011-11-18

Highlights: Black-Right-Pointing-Pointer Force via integrins or cadherins induces similar cell stiffening responses. Black-Right-Pointing-Pointer Force via integrins but not cadherins induces cell spreading. Black-Right-Pointing-Pointer Force via integrins but not cadherins induces differentiation of embryonic stem cells. -- Abstract: Increasing evidence suggests that mechanical factors play a critical role in fate decisions of stem cells. Recently we have demonstrated that a local force applied via Arg-Gly-Asp (RGD) peptides coated magnetic beads to mouse embryonic stem (ES) cells increases cell spreading and cell stiffness and decreases Oct3/4 (Pou5f1) gene expression. However, it is not clear whether the effects of the applied stress on these functions of ES cells can be extended to natural extracellular matrix proteins or cell-cell adhesion molecules. Here we show that a local cyclic shear force applied via fibronectin or laminin to integrin receptors increased cell spreading and stiffness, downregulated Oct3/4 gene expression, and decreased cell proliferation rate. In contrast, the same cyclic force applied via cell-cell adhesion molecule E-cadherin (Cdh1) had no effects on cell spreading, Oct3/4 gene expression, and the self-renewal of mouse ES cells, but induced significant cell stiffening. Our findings demonstrate that biological responses of ES cells to force applied via integrins are different from those to force via E-cadherin, suggesting that mechanical forces might play different roles in different force transduction pathways to shape early embryogenesis.

Characterization of gas tunnel type plasma sprayed hydroxyapatite-nanostructure titania composite coatings

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Yugeswaran, S.; Kobayashi, A.; Ucisik, A. Hikmet; Subramanian, B.

2015-08-01

Hydroxyapatite (HA) can be coated onto metal implants as a ceramic biocompatible coating to bridge the growth between implants and human tissue. Meanwhile many efforts have been made to improve the mechanical properties of the HA coatings without affecting its bioactivity. In the present study, nanostructure titania (TiO2) was mixed with HA powder and HA-nanostructure TiO2 composite coatings were produced by gas tunnel type plasma spraying torch under optimized spraying conditions. For this purpose, composition of 10 wt% TiO2 + 90 wt% HA, 20 wt% TiO2 + 80 wt% HA and 30 wt% TiO2 + 70 wt% HA were selected as the feedstock materials. The phase, microstructure and mechanical properties of the coatings were characterized. The obtained results validated that the increase in weight percentage of nanostructure TiO2 in HA coating significantly increased the microhardness, adhesive strength and wear resistance of the coatings. Analysis of the in vitro bioactivity and cytocompatibility of the coatings were done using conventional simulated body fluid (c-SBF) solution and cultured green fluorescent protein (GFP) labeled marrow stromal cells (MSCs) respectively. The bioactivity results revealed that the composite coating has bio-active surface with good cytocompatibility.

Innovative micro-textured hydroxyapatite and poly(l-lactic)-acid polymer composite film as a flexible, corrosion resistant, biocompatible, and bioactive coating for Mg implants.

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Kim, Sae-Mi; Kang, Min-Ho; Kim, Hyoun-Ee; Lim, Ho-Kyung; Byun, Soo-Hwan; Lee, Jong-Ho; Lee, Sung-Mi

2017-12-01

The utility of a novel ceramic/polymer-composite coating with a micro-textured microstructure that would significantly enhance the functions of biodegradable Mg implants is demonstrated here. To accomplish this, bioactive hydroxyapatite (HA) micro-dots can be created by immersing a Mg implant with a micro-patterned photoresist surface in an aqueous solution containing calcium and phosphate ions. The HA micro-dots can then be surrounded by a flexible poly(l-lactic)-acid (PLLA) polymer using spin coating to form a HA/PLLA micro-textured coating layer. The HA/PLLA micro-textured coating layer showed an excellent corrosion resistance when it was immersed in a simulated body fluid (SBF) solution and good biocompatibility, which was assessed by in vitro cell tests. In addition, the HA/PLLA micro-textured coating layer had high deformation ability, where no apparent changes in the coating layer were observed even after a 5% elongation, which would be unobtainable using HA and PLLA coating layers; furthermore, this allowed the mechanically-strained Mg implant with the HA/PLLA micro-textured coating layer to preserve its excellent corrosion resistance and biocompatibility in vitro. Copyright © 2017 Elsevier B.V. All rights reserved.

Plasma-sprayed CaTiSiO5 ceramic coating on Ti-6Al-4V with excellent bonding strength, stability and cellular bioactivity

Science.gov (United States)

Wu, Chengtie; Ramaswamy, Yogambha; Liu, Xuanyong; Wang, Guocheng; Zreiqat, Hala

2008-01-01

Novel Ca-Si-Ti-based sphene (CaTiSiO5) ceramics possess excellent chemical stability and cytocompatibility. The aim of this study was to prepare sphene coating on titanium alloy (Ti-6Al-4V) for orthopaedic applications using the plasma spray method. The phase composition, surface and interface microstructure, coating thickness, surface roughness and bonding strength of the plasma-sprayed sphene coating were analysed using X-ray diffraction, scanning electron microscopy, atomic force microscopy and the standard mechanical testing of the American Society for Testing and Materials, respectively. The results indicated that sphene coating was obtained with a uniform and dense microstructure at the interface of the Ti-6Al-4V surface and the thickness and surface roughness of the coating were approximately 150 and 10 μm, respectively. Plasma-sprayed sphene coating on Ti-6Al-4V possessed a significantly improved bonding strength and chemical stability compared with plasma-sprayed hydroxyapatite (HAp) coating. Plasma-sprayed sphene coating supported human osteoblast-like cell (HOB) attachment and significantly enhanced HOB proliferation and differentiation compared with plasma-sprayed HAp coating and uncoated Ti-6Al-4V. Taken together, plasma-sprayed sphene coating on Ti-6Al-4V possessed excellent bonding strength, chemical stability and cellular bioactivity, indicating its potential application for orthopaedic implants. PMID:18664431

Nanomodified Peek Dental Implants: Bioactive Composites and Surface Modification—A Review

Directory of Open Access Journals (Sweden)

Shariq Najeeb

2015-01-01

Full Text Available Purpose. The aim of this review is to summarize and evaluate the relevant literature regarding the different ways how polyetheretherketone (PEEK can be modified to overcome its limited bioactivity, and thereby making it suitable as a dental implant material. Study Selection. An electronic literature search was conducted via the PubMed and Google Scholar databases using the keywords “PEEK dental implants,” “nano,” “osseointegration,” “surface treatment,” and “modification.” A total of 16 in vivo and in vitro studies were found suitable to be included in this review. Results. There are many viable methods to increase the bioactivity of PEEK. Most methods focus on increasing the surface roughness, increasing the hydrophilicity and coating osseoconductive materials. Conclusion. There are many ways in which PEEK can be modified at a nanometer level to overcome its limited bioactivity. Melt-blending with bioactive nanoparticles can be used to produce bioactive nanocomposites, while spin-coating, gas plasma etching, electron beam, and plasma-ion immersion implantation can be used to modify the surface of PEEK implants in order to make them more bioactive. However, more animal studies are needed before these implants can be deemed suitable to be used as dental implants.

Multilayer bioactive glass/zirconium titanate thin films in bone tissue engineering and regenerative dentistry

Directory of Open Access Journals (Sweden)

Mozafari M

2013-04-01

Full Text Available Masoud Mozafari,1,2 Erfan Salahinejad,1,3 Vahid Shabafrooz,1 Mostafa Yazdimamaghani,1 Daryoosh Vashaee,4 Lobat Tayebi1,5 1Helmerich Advanced Technology Research Center, School of Materials Science and Engineering, Oklahoma State University, Tulsa, OK, USA; 2Biomaterials Group, Faculty of Biomedical Engineering (Center of Excellence, Amirkabir University of Technology, Tehran, Iran; 3Department of Materials Science and Engineering, School of Engineering, Shiraz University, Shiraz, Iran; 4Helmerich Advanced Technology Research Center, School of Electrical and Computer Engineering, Oklahoma State University, Tulsa, OK, USA; 5School of Chemical Engineering, Oklahoma State University, Tulsa, OK, USA Abstract: Surface modification, particularly coatings deposition, is beneficial to tissue-engineering applications. In this work, bioactive glass/zirconium titanate composite thin films were prepared by a sol-gel spin-coating method. The surface features of the coatings were studied by scanning electron microscopy, atomic force microscopy, and spectroscopic reflection analyses. The results show that uniform and sound multilayer thin films were successfully prepared through the optimization of the process variables and the application of carboxymethyl cellulose as a dispersing agent. Also, it was found that the thickness and roughness of the multilayer coatings increase nonlinearly with increasing the number of the layers. This new class of nanocomposite coatings, comprising the bioactive and inert components, is expected not only to enhance bioactivity and biocompatibility, but also to protect the surface of metallic implants against wear and corrosion. Keywords: bioactive glass, zirconium titanate, spin-coating, microstructural properties, bone/dental applications, tissue engineering

Development of strontium and magnesium substituted porous hydroxyapatite/poly(3,4-ethylenedioxythiophene) coating on surgical grade stainless steel and its bioactivity on osteoblast cells.

Science.gov (United States)

Gopi, D; Ramya, S; Rajeswari, D; Surendiran, M; Kavitha, L

2014-02-01

The present study deals with the successful development of bilayer coatings by electropolymerisation of poly(3,4-ethylenedioxythiophene) (PEDOT) on surgical grade stainless steel (316L SS) followed by the electrodeposition of strontium (Sr) and magnesium (Mg) substituted porous hydroxyapatite (Sr, Mg-HA). The bilayer coatings were characterised by Fourier transform infrared spectroscopy (FT-IR) and high resolution scanning electron microscopy (HRSEM). Corrosion resistance of the obtained coatings was investigated in Ringer's solution by electrochemical techniques and the results were in good agreement with those obtained from chemical analysis, namely inductively coupled plasma atomic emission spectrometry (ICP-AES). Also, the mechanical and biological properties of the bilayer coatings were analyzed. From the obtained results it was evident that the PEDOT/Sr, Mg-HA bilayer exhibited greater adhesion strength than the Sr, Mg-HA coated 316L SS. In vitro cell adhesion test of the Sr, Mg-HA coating on PEDOT coated specimen is found to be more bioactive compared to that of the single substituted hydroxyapatite (Sr or Mg-HA) on the PEDOT coated 316L SS. Thus, the PEDOT/Sr, Mg-HA bilayer coated 316L SS can serve as a prospective implant material for biomedical applications. Copyright © 2013 Elsevier B.V. All rights reserved.

Porous biomorphic silicon carbide ceramics coated with hydroxyapatite as prospective materials for bone implants.

Science.gov (United States)

Gryshkov, Oleksandr; Klyui, Nickolai I; Temchenko, Volodymyr P; Kyselov, Vitalii S; Chatterjee, Anamika; Belyaev, Alexander E; Lauterboeck, Lothar; Iarmolenko, Dmytro; Glasmacher, Birgit

2016-11-01

Porous and cytocompatible silicon carbide (SiC) ceramics derived from wood precursors and coated with bioactive hydroxyapatite (HA) and HA-zirconium dioxide (HA/ZrO2) composite are materials with promising application in engineering of bone implants due to their excellent mechanical and structural properties. Biomorphic SiC ceramics have been synthesized from wood (Hornbeam, Sapele, Tilia and Pear) using a forced impregnation method. The SiC ceramics have been coated with bioactive HA and HA/ZrO2 using effective gas detonation deposition approach (GDD). The surface morphology and cytotoxicity of SiC ceramics as well as phase composition and crystallinity of deposited coatings were analyzed. It has been shown that the porosity and pore size of SiC ceramics depend on initial wood source. The XRD and FTIR studies revealed the preservation of crystal structure and phase composition of in the HA coating, while addition of ZrO2 to the initial HA powder resulted in significant decomposition of the final HA/ZrO2 coating and formation of other calcium phosphate phases. In turn, NIH 3T3 cells cultured in medium exposed to coated and uncoated SiC ceramics showed high re-cultivation efficiency as well as metabolic activity. The recultivation efficiency of cells was the highest for HA-coated ceramics, whereas HA/ZrO2 coating improved the recultivation efficiency of cells as compared to uncoated SiC ceramics. The GDD method allowed generating homogeneous HA coatings with no change in calcium to phosphorus ratio. In summary, porous and cytocompatible bio-SiC ceramics with bioactive coatings show a great promise in construction of light, robust, inexpensive and patient-specific bone implants for clinical application. Copyright © 2016 Elsevier B.V. All rights reserved.

Novel Development of Biocompatible Coatings for Bone Implants

Directory of Open Access Journals (Sweden)

Nicholas Yue Hou

2015-10-01

Full Text Available Prolonged life expectancy also results in an increased need for high-performance orthopedic implants. It has been shown that a compromised tissue-implant interface could lead to adverse immune-responses and even the dislodging of the implant. To overcome these obstacles, our research team has been seeking ways to decrease the risk of faulty tissue-implant interfaces by improving the biocompatibility and the osteo-inductivity of conventional orthopedic implants using ultrafine particle coatings. These particles were enriched with various bioactive additives prior to coating, and the coated biomaterial surfaces exhibited significantly increased biocompatibility and osteoinductivity. Physical assessments firstly confirmed the proper incorporation of the bioactive additives after examining their surface chemical composition. Then, in vitro assays demonstrated the biocompatibility and osteo-inductivity of the coated surfaces by studying the morphology of attached cells and their mineralization abilities. In addition, by quantifying the responses, activities and gene expressions, cellular evaluations confirmed the positive effects of these polymer based bioactive coatings. Consequently, the bioactive ultrafine polymer particles demonstrated their ability in improving the biocompatibility and osteo-inductivity of conventional orthopedic implants. As a result, our research team hope to apply this technology to the field of orthopedic implants by making them more effective medical devices through decreasing the risk of implant-induced immune responses and the loosening of the implant.

Xerogel Interfaced Nanofibers Stimulate Bone Regeneration Through the Activation of Integrin and Bone Morphogenetic Protein Pathways.

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Lee, Yoo-Mi; Yun, Hyung-Mun; Lee, Hye-Young; Lim, Hyun-Chang; Lee, Hae-Hyoung; Kim, Hae-Won; Kim, Eun-Cheol

2017-02-01

A xerogel was interfaced onto biopolymer nanofibers though a core–shell electrospinning design for bone regeneration. The xerogel-interfaced biopolymer nanofibrous matrix was bioactive and highly hydrophilic, with a significant decrease in the water contact angle. The matrix showed excellent in vitro responses of primary osteoblasts in terms of adhesion, proliferation, and migration. Furthermore, the osteoblastic differentiation of cells, including alkaline phosphatase activity, mineralization, and gene expression, was significantly upregulated by the xerogel interface. In vivo animal tests in a critical-sized calvarial defect confirmed the new bone formation ability of the xerogel-surfaced nanofiber matrices. The underlying signaling mechanisms of the stimulation were implied to be integrin and bone morphogenetic protein (BMP) pathways, as demonstrated by the activation of integrin (α2β1) and downstream signaling molecules (FAK, paxillin, RhoA, MAPK, and NF-κB), as well as the BMPs and the downstream transcription factor Smad1/5/8. Taking these findings together, the xerogel-surfaced biopolymer nanofibers are proposed to be a promising scaffold candidate for bone regeneration.

Pregnancy-specific glycoproteins bind integrin αIIbβ3 and inhibit the platelet-fibrinogen interaction.

Science.gov (United States)

Shanley, Daniel K; Kiely, Patrick A; Golla, Kalyan; Allen, Seamus; Martin, Kenneth; O'Riordan, Ronan T; Ball, Melanie; Aplin, John D; Singer, Bernhard B; Caplice, Noel; Moran, Niamh; Moore, Tom

2013-01-01

Pregnancy-specific glycoproteins (PSGs) are immunoglobulin superfamily members encoded by multigene families in rodents and primates. In human pregnancy, PSGs are secreted by the syncytiotrophoblast, a fetal tissue, and reach a concentration of up to 400 ug/ml in the maternal bloodstream at term. Human and mouse PSGs induce release of anti-inflammatory cytokines such as IL-10 and TGFβ1 from monocytes, macrophages, and other cell types, suggesting an immunoregulatory function. RGD tri-peptide motifs in the majority of human PSGs suggest that they may function like snake venom disintegrins, which bind integrins and inhibit interactions with ligands. We noted that human PSG1 has a KGD, rather than an RGD motif. The presence of a KGD in barbourin, a platelet integrin αIIbβ3 antagonist found in snake venom, suggested that PSG1 may be a selective αIIbβ3 ligand. Here we show that human PSG1 binds αIIbβ3 and inhibits the platelet - fibrinogen interaction. Unexpectedly, however, the KGD is not critical as multiple PSG1 domains independently bind and inhibit αIIbβ3 function. Human PSG9 and mouse Psg23 are also inhibitory suggesting conservation of this function across primate and rodent PSG families. Our results suggest that in species with haemochorial placentation, in which maternal blood is in direct contact with fetal trophoblast, the high expression level of PSGs reflects a requirement to antagonise abundant (3 mg/ml) fibrinogen in the maternal circulation, which may be necessary to prevent platelet aggregation and thrombosis in the prothrombotic maternal environment of pregnancy.

Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells†

Science.gov (United States)

Das, Lipsa; Anderson, Todd A.; Gard, Jaime M.C.; Sroka, Isis C.; Strautman, Stephanie R.; Nagle, Raymond B.; Morrissey, Colm; Knudsen, Beatrice S.; Cress, Anne E.

2017-01-01

Laminin binding integrins α6 (CD49f) and α3 (CD49c) are persistently but differentially expressed in prostate cancer (PCa). Integrin internalization is an important determinant of their cell surface expression and function. Using flow cytometry, and first order kinetic modelling, we quantitated the intrinsic internalization rates of integrin subunits in a single cycle of internalization. In PCa cell line DU145, α6 integrin internalized with a rate constant (kactual) of 3.25min−1, 3-fold faster than α3 integrin (1.0 min−1), 1.5-fold faster than the vitronectin binding αv integrin (CD51) (2.2 min−1), and significantly slower than the unrelated transferrin receptor (CD71) (15 min−1). Silencing of α3 integrin protein expression in DU145, PC3 and PC3B1 cells resulted in up to a 1.71-fold increase in kactual for α6 integrin. The internalized α6 integrin was targeted to early endosomes but not to lamp1 vesicles. Depletion of α3 integrin expression resulted in redistribution of α6β4 integrin to an observed cell-cell staining pattern that is consistent with a suprabasal distribution observed in epidermis and early PIN lesions in PCa. Depletion of α3 integrin increased cell migration by 1.8 fold, which was dependent on α6β1 integrin. Silencing of α6 integrin expression however, had no significant effect on the kactual of α3 integrin or its distribution in early endosomes. These results indicate that α3 and α6 integrins have significantly different internalization kinetics and that coordination exists between them for internalization. This article is protected by copyright. All rights reserved PMID:27509031

Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells.

Science.gov (United States)

Das, Lipsa; Anderson, Todd A; Gard, Jaime M C; Sroka, Isis C; Strautman, Stephanie R; Nagle, Raymond B; Morrissey, Colm; Knudsen, Beatrice S; Cress, Anne E

2017-05-01

Laminin binding integrins α6 (CD49f) and α3 (CD49c) are persistently but differentially expressed in prostate cancer (PCa). Integrin internalization is an important determinant of their cell surface expression and function. Using flow cytometry, and first order kinetic modeling, we quantitated the intrinsic internalization rates of integrin subunits in a single cycle of internalization. In PCa cell line DU145, α6 integrin internalized with a rate constant (k actual ) of 3.25 min -1 , threefold faster than α3 integrin (1.0 min -1 ), 1.5-fold faster than the vitronectin binding αv integrin (CD51) (2.2 min -1 ), and significantly slower than the unrelated transferrin receptor (CD71) (15 min -1 ). Silencing of α3 integrin protein expression in DU145, PC3, and PC3B1 cells resulted in up to a 1.71-fold increase in k actual for α6 integrin. The internalized α6 integrin was targeted to early endosomes but not to lamp1 vesicles. Depletion of α3 integrin expression resulted in redistribution of α6β4 integrin to an observed cell-cell staining pattern that is consistent with a suprabasal distribution observed in epidermis and early PIN lesions in PCa. Depletion of α3 integrin increased cell migration by 1.8-fold, which was dependent on α6β1 integrin. Silencing of α6 integrin expression however, had no significant effect on the k actual of α3 integrin or its distribution in early endosomes. These results indicate that α3 and α6 integrins have significantly different internalization kinetics and that coordination exists between them for internalization. J. Cell. Biochem. 118: 1038-1049, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Effect of implant design and bioactive glass coating on biomechanical properties of fiber-reinforced composite implants.

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Ballo, Ahmed M; Akca, Eralp; Ozen, Tuncer; Moritz, Niko; Lassila, Lippo; Vallittu, Pekka; Närhi, Timo

2014-08-01

This study aimed to evaluate the influence of implant design and bioactive glass (BAG) coating on the response of bone to fiber-reinforced composite (FRC) implants. Three different FRC implant types were manufactured for the study: non-threaded implants with a BAG coating; threaded implants with a BAG coating; and threaded implants with a grit-blasted surface. Thirty-six implants (six implants for each group per time point) were installed in the tibiae of six pigs. After an implantation period of 4 and 12 wk, the implants were retrieved and prepared for micro-computed tomography (micro-CT), push-out testing, and scanning electron microscopy analysis. Micro-CT demonstrated that the screw-threads and implant structure remained undamaged during the installation. The threaded FRC/BAG implants had the highest bone volume after 12 wk of implantation. The push-out strengths of the threaded FRC/BAG implants after 4 and 12 wk (463°N and 676°N, respectively) were significantly higher than those of the threaded FRC implants (416°N and 549°N, respectively) and the nonthreaded FRC/BAG implants (219°N and 430°N, respectively). Statistically significant correlation was found between bone volume and push-out strength values. This study showed that osseointegrated FRC implants can withstand the static loading up to failure without fracture, and that the addition of BAG significantly improves the push-out strength of FRC implants. © 2014 Eur J Oral Sci.

Thermal spraying of functionally graded calcium phosphate coatings for biomedical implants

Science.gov (United States)

Wang, Y.; Khor, K. A.; Cheang, P.

1998-03-01

Biomedical requirements in a prosthesis are often complex and diverse in nature. Biomaterials for implants have to display a wide range of adaptability to suit the various stages of the bio-integration process of any foreign material into the human body. Often, a combination of materials is needed. The preparation of a functionally graded bioceramic coating composed of essentially calcium phosphate compounds is explored. The coating is graded in accordance to adhesive strength, bioactivity, and bioresorbability. The bond coat on the Ti-6Al-4V stub is deposited with a particle range of the hydroxyapatite (HA) that will provide a high adhesive strength and bioactivity but have poor bioresorption properties. The top coat, however, is composed of predominantly α-tricalcium phosphate (α-TCP) that is highly bioresorbable. This arrangement has the propensity of allowing accelerated bio-integration of the coating by the body tissues as the top layer is rapidly resorbed, leaving the more bioactive intermediate layer to facilitate the much needed bioactive properties for proper osteoconduction. The processing steps and problems are highlighted, as well as the results of post-spray heat treatment.

Galectin-3 induces clustering of CD147 and integrin-β1 transmembrane glycoprotein receptors on the RPE cell surface.

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Claudia S Priglinger

Full Text Available Proliferative vitreoretinopathy (PVR is a blinding disease frequently occurring after retinal detachment surgery. Adhesion, migration and matrix remodeling of dedifferentiated retinal pigment epithelial (RPE cells characterize the onset of the disease. Treatment options are still restrained and identification of factors responsible for the abnormal behavior of the RPE cells will facilitate the development of novel therapeutics. Galectin-3, a carbohydrate-binding protein, was previously found to inhibit attachment and spreading of retinal pigment epithelial cells, and thus bares the potential to counteract PVR-associated cellular events. However, the identities of the corresponding cell surface glycoprotein receptor proteins on RPE cells are not known. Here we characterize RPE-specific Gal-3 containing glycoprotein complexes using a proteomic approach. Integrin-β1, integrin-α3 and CD147/EMMPRIN, a transmembrane glycoprotein implicated in regulating matrix metalloproteinase induction, were identified as potential Gal-3 interactors on RPE cell surfaces. In reciprocal immunoprecipitation experiments we confirmed that Gal-3 associated with CD147 and integrin-β1, but not with integrin-α3. Additionally, association of Gal-3 with CD147 and integrin-β1 was observed in co-localization analyses, while integrin-α3 only partially co-localized with Gal-3. Blocking of CD147 and integrin-β1 on RPE cell surfaces inhibited binding of Gal-3, whereas blocking of integrin-α3 failed to do so, suggesting that integrin-α3 is rather an indirect interactor. Importantly, Gal-3 binding promoted pronounced clustering and co-localization of CD147 and integrin-β1, with only partial association of integrin-α3. Finally, we show that RPE derived CD147 and integrin-β1, but not integrin-α3, carry predominantly β-1,6-N-actyl-D-glucosamine-branched glycans, which are high-affinity ligands for Gal-3. We conclude from these data that extracellular Gal-3 triggers

Galectin-3 Induces Clustering of CD147 and Integrin-β1 Transmembrane Glycoprotein Receptors on the RPE Cell Surface

Science.gov (United States)

Priglinger, Claudia S.; Szober, Christoph M.; Priglinger, Siegfried G.; Merl, Juliane; Euler, Kerstin N.; Kernt, Marcus; Gondi, Gabor; Behler, Jennifer; Geerlof, Arie; Kampik, Anselm; Ueffing, Marius; Hauck, Stefanie M.

2013-01-01

Proliferative vitreoretinopathy (PVR) is a blinding disease frequently occurring after retinal detachment surgery. Adhesion, migration and matrix remodeling of dedifferentiated retinal pigment epithelial (RPE) cells characterize the onset of the disease. Treatment options are still restrained and identification of factors responsible for the abnormal behavior of the RPE cells will facilitate the development of novel therapeutics. Galectin-3, a carbohydrate-binding protein, was previously found to inhibit attachment and spreading of retinal pigment epithelial cells, and thus bares the potential to counteract PVR-associated cellular events. However, the identities of the corresponding cell surface glycoprotein receptor proteins on RPE cells are not known. Here we characterize RPE-specific Gal-3 containing glycoprotein complexes using a proteomic approach. Integrin-β1, integrin-α3 and CD147/EMMPRIN, a transmembrane glycoprotein implicated in regulating matrix metalloproteinase induction, were identified as potential Gal-3 interactors on RPE cell surfaces. In reciprocal immunoprecipitation experiments we confirmed that Gal-3 associated with CD147 and integrin-β1, but not with integrin-α3. Additionally, association of Gal-3 with CD147 and integrin-β1 was observed in co-localization analyses, while integrin-α3 only partially co-localized with Gal-3. Blocking of CD147 and integrin-β1 on RPE cell surfaces inhibited binding of Gal-3, whereas blocking of integrin-α3 failed to do so, suggesting that integrin-α3 is rather an indirect interactor. Importantly, Gal-3 binding promoted pronounced clustering and co-localization of CD147 and integrin-β1, with only partial association of integrin-α3. Finally, we show that RPE derived CD147 and integrin-β1, but not integrin-α3, carry predominantly β-1,6-N-actyl-D-glucosamine-branched glycans, which are high-affinity ligands for Gal-3. We conclude from these data that extracellular Gal-3 triggers clustering of CD147 and

Hydroxyapatite coatings for biomedical applications

CERN Document Server

Zhang, Sam

2013-01-01

Hydroxyapatite coatings are of great importance in the biological and biomedical coatings fields, especially in the current era of nanotechnology and bioapplications. With a bonelike structure that promotes osseointegration, hydroxyapatite coating can be applied to otherwise bioinactive implants to make their surface bioactive, thus achieving faster healing and recovery. In addition to applications in orthopedic and dental implants, this coating can also be used in drug delivery. Hydroxyapatite Coatings for Biomedical Applications explores developments in the processing and property characteri

Mechanically enforced bond dissociation reports synergistic influence of Mn2+ and Mg2+ on the interaction between integrin α7β1 and invasin

DEFF Research Database (Denmark)

Ligezowska, Agnieszka; Boye, Kristian; Eble, Johannes A.

2011-01-01

addition of these cations to the measurement buffer, we observe a pronounced increase in the force necessary to separate integrin and invasin coated beads. Both ions were found to work synergistically. With free invasin in the measurement buffer we furthermore observe that competitive blocking of binding...... sites overrides the increase in binding strength of individual beads. We show that this is due to a very strong dependence of bond affinity on divalent ions. Our study illustrates the importance of divalent ions for the regulation of force transmission by integrin ligand bonds on the molecular level...

Bacterial biofilm formation versus mammalian cell growth on titanium-based mono- and bi-functional coating

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G Subbiahdoss

2010-05-01

Full Text Available Biomaterials-associated-infections (BAI are serious complications in modern medicine. Although non-adhesive coatings, like polymer-brush coatings, have been shown to prevent bacterial adhesion, they do not support cell growth. Bi-functional coatings are supposed to prevent biofilm formation while supporting tissue integration. Here, bacterial and cellular responses to poly(ethylene glycol (PEG brush-coatings on titanium oxide presenting the integrin-active peptide RGD (arginine-glycine-aspartic acid (bioactive “PEG-RGD” were compared to mono-functional PEG brush-coatings (biopassive “PEG” and bare titanium oxide (TiO2 surfaces under flow. Staphylococcus epidermidis ATCC 35983 was deposited on the surfaces under a shear rate of 11 s-1 for 2 h followed by seeding of U2OS osteoblasts. Subsequently, both S. epidermidis and U2OS cells were grown simultaneously on the surfaces for 48 h under low shear (0.14 s-1. After 2 h, staphylococcal adhesion was reduced to 3.6±1.8 × 103 and 6.0±3.9 × 103 cm-2 on PEG and PEG-RGD coatings respectively, compared to 1.3±0.4 × 105 cm-2 for the TiO2 surface. When allowed to grow for 48 h, biofilms formed on all surfaces. However, biofilms detached from the PEG and PEG-RGD coatings when exposed to an elevated shear (5.6 s-1 U2OS cells neither adhered nor spread on PEG brush-coatings, regardless of the presence of biofilm. In contrast, in the presence of biofilm, U2OS cells adhered and spread on PEG-RGD coatings with a significantly higher surface coverage than on bare TiO2. The detachment of biofilm and the high cell surface coverage revealed the potential significance of PEG-RGD coatings in the context of the “race for the surface” between bacteria and mammalian cells.

Syndecans promote integrin-mediated adhesion of mesenchymal cells in two distinct pathways

DEFF Research Database (Denmark)

Whiteford, James; Behrends, Volker; Kirby, Hishani

2007-01-01

and signaling through the cytoplasmic domain of syndecan-4. Here an alternate pathway mediated by the extracellular domains of syndecans-2 and -4 is characterized that is independent of both heparan sulphate and syndecan signaling. This pathway is restricted to mesenchymal cells and was not seen in any...... epithelial cell line tested, apart from vascular endothelia. The syndecan ectodomains coated as substrates promoted integrin-dependent attachment, spreading and focal adhesion formation. Syndecan-4 null cells were competent, as were fibroblasts compromised in heparan sulphate synthesis that were unable...

Different Phenotypes in Human Prostate Cancer: α6 or α3 Integrin in Cell-extracellular Adhesion Sites

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Monika Schmelz

2002-01-01

Full Text Available The distribution of α6/α3 integrin in adhesion complexes at the basal membrane in human normal and cancer prostate glands was analyzed in 135 biopsies from 61 patients. The levels of the polarized α6/α3 integrin expression at the basal membrane of prostate tumor glands were determined by quantitative immunohistochemistry. The α6/α3 integrin expression was compared with Gleason sum score, pathological stage, and preoperative serum prostate-specific antigen (PSA. The associations were assessed by statistical methods. Eighty percent of the tumors expressed the α6 or α3 integrin and 20% was integrin-negative. Gleason sum score, but not serum PSA, was associated with the integrin expression. Low Gleason sum score correlated with increased integrin expression, high Gleason sum score with low and negative integrin expression. Three prostate tumor phenotypes were distinguished based on differential integrin expression. Type I coexpressed both α6 and α3 subunits, type II exclusively expressed a6 integrin, and type III expressed α3 integrin only. Fifteen cases were further examined for the codistribution of vinculin, paxillin, and CD 151 on frozen serial sections using confocal laser scanning microscopy. The α6/α3 integrins, CD151, paxillin, and vinculin were present within normal glands. In prostate carcinoma, α6 integrin was colocalized with CD 151, but not with vinculin or paxillin. In tumor phenotype I, the α6 subunit did not colocalize with the α3 subunit indicating the existence of two different adhesion complexes. Human prostate tumors display on their cell surface the α6β1 and/or α3β1 integrins. Three tumor phenotypes associated with two different adhesion complexes were identified, suggesting a reorganization of cell adhesion structures in prostate cancer.

Fabrication and characterization of modified-hydroxyapatite/polyetheretherketone coating materials

Energy Technology Data Exchange (ETDEWEB)

Ma, Rui [College of Optoelectronic Engineering, Shenzhen University, Shenzhen 518060 (China); College of Chemistry and Chemical Engineering, Shenzhen University, Shenzhen 518060 (China); Fang, Lin, E-mail: fanglinhit@163.com [College of Chemistry and Chemical Engineering, Shenzhen University, Shenzhen 518060 (China); Luo, Zhongkuan [College of Chemistry and Chemical Engineering, Shenzhen University, Shenzhen 518060 (China); Zheng, Ruisheng [College of Optoelectronic Engineering, Shenzhen University, Shenzhen 518060 (China); Song, Shenhua; Weng, Luqian; Lei, JinPing [Materials Science and Engineering, Shenzhen Graduate School, Harbin Institute of Technology, Shenzhen 518055 (China)

2014-09-30

Highlights: • 45 wt%-Hydroxyaptite/polyetheretherketone (HA/PEEK) coating materials modified by silane coupling agent (KH560) on PEEK substrate is successfully fabricated by solution casting method. • Strong bonding between the composite coating and the PEEK substrate is achieved. • HA/PEEK coating materials exhibit better bioactivity. - Abstract: 45 wt%-Hydroxyaptite/polyetheretherketone (HA/PEEK) coating materials modified by silane coupling agent (KH560) on PEEK substrate were successfully fabricated by solution casting method and characterized by Fourier transform infrared spectroscopy (FTIR), X-Ray Diffraction (XRD), scanning electron microscopy (SEM), and tensile testing. The modified HA fillers were obtained to be uniformly distributed in the HA/PEEK coating, which has better properties of tensile strength and fracture toughness than those of the unmodified specimen. A good bonding between the composite coating and the PEEK substrate was achieved by solution casting method, resulting in integral-fracture without falling apart or delaminating during tensile loading. The modified specimens dipped into simulated body fluid (SBF) were characterized by SEM, XRD and FTIR, indicating that the bioactivity of the dipped materials was demonstrated more apparent with extending the dipping time. Therefore, the coating materials may become the substitutes for the hard tissues of the human body in the future, which could realize the balance between the mechanical properties and the bioactivity by modifying the structural design of the coating.

Fabrication and characterization of modified-hydroxyapatite/polyetheretherketone coating materials

International Nuclear Information System (INIS)

Ma, Rui; Fang, Lin; Luo, Zhongkuan; Zheng, Ruisheng; Song, Shenhua; Weng, Luqian; Lei, JinPing

2014-01-01

Highlights: • 45 wt%-Hydroxyaptite/polyetheretherketone (HA/PEEK) coating materials modified by silane coupling agent (KH560) on PEEK substrate is successfully fabricated by solution casting method. • Strong bonding between the composite coating and the PEEK substrate is achieved. • HA/PEEK coating materials exhibit better bioactivity. - Abstract: 45 wt%-Hydroxyaptite/polyetheretherketone (HA/PEEK) coating materials modified by silane coupling agent (KH560) on PEEK substrate were successfully fabricated by solution casting method and characterized by Fourier transform infrared spectroscopy (FTIR), X-Ray Diffraction (XRD), scanning electron microscopy (SEM), and tensile testing. The modified HA fillers were obtained to be uniformly distributed in the HA/PEEK coating, which has better properties of tensile strength and fracture toughness than those of the unmodified specimen. A good bonding between the composite coating and the PEEK substrate was achieved by solution casting method, resulting in integral-fracture without falling apart or delaminating during tensile loading. The modified specimens dipped into simulated body fluid (SBF) were characterized by SEM, XRD and FTIR, indicating that the bioactivity of the dipped materials was demonstrated more apparent with extending the dipping time. Therefore, the coating materials may become the substitutes for the hard tissues of the human body in the future, which could realize the balance between the mechanical properties and the bioactivity by modifying the structural design of the coating

Lymphocyte integrin expression differences between SIRS and sepsis patients.

Science.gov (United States)

Heffernan, D S; Monaghan, S F; Ayala, Alfred

2017-11-01

Systemic Inflammatory Response Syndrome (SIRS) and sepsis remain leading causes of death. Despite many similarities, the two entities are very distinct clinically and immunologically. T-Lymphocytes play a key pivotal role in the pathogenesis and ultimately outcome following both SIRS and sepsis. Integrins are essential in the trafficking and migration of lymphocytes. They also serve vital roles in efficient wound healing and clearance of infections. Here, we investigate whether integrin expression, specifically β1 (CD29) and β2 (CD18), are disrupted in SIRS and sepsis, and assess differences in integrin expression between these two critically ill clinical categories. T-Lymphocytes were isolated from whole blood collected from ICU patients exhibiting SIRS or sepsis. Samples were analyzed for CD18 (β2) and CD29 (β1) on CD3 + T cells through flow cytometry. Septic patients were stratified into either exclusively abdominal or non-abdominal sources of sepsis. CD18 was almost ubiquitously expressed on CD3 + T cells irrespective of clinical condition. However, CD29 (β1 integrin) was lowest in SIRS patients (20.4% of CD3 + T cells) when compared with either septic patients (35.5%) or healthy volunteers (54.1%). Furthermore, there was evidence of compartmentalization in septic patients, where abdominal sources had a greater percentage of CD3 + CD29 + T cells (41.7%) when compared with those with non-abdominal sources (29.5%). Distinct differences in T-cell integrin expression exists between patients in SIRS versus sepsis, as well as relative to the source of sepsis. Further work is needed to understand cause and effect relative to the progression from SIRS into sepsis.

COMPARISON OF SOL-GEL SILICATE COATINGS ON Ti SUBSTRATE

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DIANA HORKAVCOVÁ

2012-12-01

Full Text Available The objective of the submitted work was to prepare and to characterize two types of silicate coatings prepared by the sol-gel method using the dip-coating technique on a titanium substrate. Efforts have been made to use mechanical properties of bio-inert titanium and bioactive properties of a silicate layer enriched with an admixture of compounds identified below. The first group consisted of silicate coatings containing silver, brushite and monetite. The other group of silicate coatings contained calcium nitrate and triethyl phosphate. Mechanically and chemically treated titanium substrates were dipped into sols and dried and fired. Silicate coatings from the first group were also chemically treated in 10 mol.l-1 solution of sodium hydroxide. All coatings were measured to determine their adhesive and bioactive properties and furthermore the antibacterial properties were tested in the case of first group. Surfaces of the coated substrates were investigated after the firing and after the individual tests with optical and electron microscopy and X-ray microdiffraction. A tape test demonstrated excellent adhesive property of all coatings to the substrate, classified with degree 5. A static in vitro test demonstrated bioactivity of nearly all the coatings. The basic silicate coating from the first group and one type of coating from the second group were identified as inert. Antibacterial properties of silicate coatings containing silver showed to be different when tested against Escherichia coli bacteria. A complete inhibition of the growth of bacteria under our experimental conditions was observed for the coating containing silver and monetite and a partial inhibition of the growth of bacteria for coatings containing silver and silver in combination with brushite.

Layered titanates with fibrous nanotopographic features as reservoir for bioactive ions to enhance osteogenesis

Science.gov (United States)

Song, Xiaoxia; Tang, Wei; Gregurec, Danijela; Yate, Luis; Moya, Sergio Enrique; Wang, Guocheng

2018-04-01

In this study, an osteogenic environment was constructed on Ti alloy implants by in-situ formation of nanosized fibrous titanate, Na2Ti6O13, loaded with bioactive ions, i.e. Sr, Mg and Zn, to enhance surface bioactivity. The bioactive ions were loaded by ion exchange with sodium located at inter-layer positions between the TiO6 slabs, and their release was not associated with the degradation of the structural unit of the titanate. In-vitro cell culture experiments using MC3T3-E1 cells proved that both bioactive ions and nanotopographic features are critical in promoting osteogenic differentiation of the cells. It was found that the osteogenic functions of the titanate can be modulated by the type and amount of ions incorporated. This study points out that nanosized fibrous titanate formed on the Ti alloy can be a promising reservoir for bioactive ions. The major advantage of this approach over other alternatives for bioactive ion delivery using degradable bioceramic coatings is its capacity of maintaining the structural integrity of the coating and thus avoiding structural deterioration and potential mechanical failure.

Novel nanocomposite coating for dental implant applications in vitro and in vivo evaluation.

Science.gov (United States)

Mehdikhani-Nahrkhalaji, M; Fathi, M H; Mortazavi, V; Mousavi, S B; Hashemi-Beni, B; Razavi, S M

2012-02-01

This study aimed at preparation and in vitro and in vivo evaluation of novel bioactive, biodegradable, and antibacterial nanocomposite coating for the improvement of stem cells attachment and antibacterial activity as a candidate for dental implant applications. Poly (lactide-co-glycolide)/bioactive glass/hydroxyapatite (PBGHA) nanocomposite coating was prepared via solvent casting process. The nanoparticle amounts of 10, 15, and 20 weight percent (wt%) were chosen in order to determine the optimum amount of nanoparticles suitable for preparing an uniform coating. Bioactivity and degradation of the coating with an optimum amount of nanoparticles were evaluated by immersing the prepared samples in simulated body fluid and phosphate buffer saline (PBS), respectively. The effect of nanocomposite coating on the attachment and viability of human adipose-derived stem cells (hASCs) was investigated. Kirschner wires (K-wires) of stainless steel were coated with the PBGHA nanocomposite coating, and mechanical stability of the coating was studied during intramedullary implantation into rabbit tibiae. The results showed that using 10 wt% nanoparticles (5 wt% HA and 5 wt% BG) in the nanocomposite could provide the desired uniform coating. The study of in vitro bioactivity showed rapid formation of bone-like apatite on the PBGHA coating. It was degraded considerably after about 60 days of immersion in PBS. The hASCs showed excellent attachment and viability on the coating. PBGHA coating remained stable on the K-wires with a minimum of 96% of the original coating mass. It was concluded that PBGHA nanocomposite coating provides an ideal surface for the stem cells attachment and viability. In addition, it could induce antibacterial activity, simultaneously.

Current Strategies to Improve the Bioactivity of PEEK

Science.gov (United States)

Ma, Rui; Tang, Tingting

2014-01-01

The synthetic thermoplastic polymer polyetheretherketone (PEEK) is becoming a popular component of clinical orthopedic and spinal applications, but its practical use suffers from several limitations. Although PEEK is biocompatible, chemically stable, radiolucent and has an elastic modulus similar to that of normal human bone, it is biologically inert, preventing good integration with adjacent bone tissues upon implantation. Recent efforts have focused on increasing the bioactivity of PEEK to improve the bone-implant interface. Two main strategies have been used to overcome the inert character of PEEK. One approach is surface modification to activate PEEK through surface treatment alone or in combination with a surface coating. Another strategy is to prepare bioactive PEEK composites by impregnating bioactive materials into PEEK substrate. Researchers believe that modified bioactive PEEK will have a wide range of orthopedic applications. PMID:24686515

The changing integrin expression and a role for integrin β8 in the chondrogenic differentiation of mesenchymal stem cells.

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Vanessa L S LaPointe

Full Text Available Many cartilage tissue engineering approaches aim to differentiate human mesenchymal stem cells (hMSCs into chondrocytes and develop cartilage in vitro by targeting cell-matrix interactions. We sought to better inform the design of cartilage tissue engineering scaffolds by understanding how integrin expression changes during chondrogenic differentiation. In three models of in vitro chondrogenesis, we studied the temporal change of cartilage phenotype markers and integrin subunits during the differentiation of hMSCs. We found that transcript expression of most subunits was conserved across the chondrogenesis models, but was significantly affected by the time-course of differentiation. In particular, ITGB8 was up-regulated and its importance in chondrogenesis was further established by a knockdown of integrin β8, which resulted in a non-hyaline cartilage phenotype, with no COL2A1 expression detected. In conclusion, we performed a systematic study of the temporal changes of integrin expression during chondrogenic differentiation in multiple chondrogenesis models, and revealed a role for integrin β8 in chondrogenesis. This work enhances our understanding of the changing adhesion requirements of hMSCs during chondrogenic differentiation and underlines the importance of integrins in establishing a cartilage phenotype.

The Changing Integrin Expression and a Role for Integrin β8 in the Chondrogenic Differentiation of Mesenchymal Stem Cells

Science.gov (United States)

LaPointe, Vanessa L. S.; Verpoorte, Amanda; Stevens, Molly M.

2013-01-01

Many cartilage tissue engineering approaches aim to differentiate human mesenchymal stem cells (hMSCs) into chondrocytes and develop cartilage in vitro by targeting cell-matrix interactions. We sought to better inform the design of cartilage tissue engineering scaffolds by understanding how integrin expression changes during chondrogenic differentiation. In three models of in vitro chondrogenesis, we studied the temporal change of cartilage phenotype markers and integrin subunits during the differentiation of hMSCs. We found that transcript expression of most subunits was conserved across the chondrogenesis models, but was significantly affected by the time-course of differentiation. In particular, ITGB8 was up-regulated and its importance in chondrogenesis was further established by a knockdown of integrin β8, which resulted in a non-hyaline cartilage phenotype, with no COL2A1 expression detected. In conclusion, we performed a systematic study of the temporal changes of integrin expression during chondrogenic differentiation in multiple chondrogenesis models, and revealed a role for integrin β8 in chondrogenesis. This work enhances our understanding of the changing adhesion requirements of hMSCs during chondrogenic differentiation and underlines the importance of integrins in establishing a cartilage phenotype. PMID:24312400

Titanium addition influences antibacterial activity of bioactive glass coatings on metallic implants.

Science.gov (United States)

Rodriguez, Omar; Stone, Wendy; Schemitsch, Emil H; Zalzal, Paul; Waldman, Stephen; Papini, Marcello; Towler, Mark R

2017-10-01

In an attempt to combat the possibility of bacterial infection and insufficient bone growth around metallic, surgical implants, bioactive glasses may be employed as coatings. In this work, silica-based and borate-based glass series were synthesized for this purpose and subsequently characterized in terms of antibacterial behavior, solubility and cytotoxicity. Borate-based glasses were found to exhibit significantly superior antibacterial properties and increased solubility compared to their silica-based counterparts, with BRT0 and BRT3 (borate-based glasses with 0 and 15 mol% of titanium dioxide incorporated, respectively) outperforming the remainder of the glasses, both borate and silicate based, in these respects. Atomic Absorption Spectroscopy confirmed the release of zinc ions (Zn 2+ ), which has been linked to the antibacterial abilities of glasses SRT0, BRT0 and BRT3, with inhibition effectively achieved at concentrations lower than 0.7 ppm. In vitro cytotoxicity studies using MC3T3-E1 osteoblasts confirmed that cell proliferation was affected by all glasses in this study, with decreased proliferation attributed to a faster release of sodium ions over calcium ions in both glass series, factor known to slow cell proliferation in vitro .

The in vitro biomineralization and cytocompatibility of polydopamine coated carbon nanotubes

International Nuclear Information System (INIS)

Yan Penghua; Wang Jinqing; Wang Lin; Liu Bin; Lei Ziqiang; Yang Shengrong

2011-01-01

In this work, polydopamine coated carbon nanotubes were firstly prepared by a simple and feasible route. Then, for comparison, the in vitro bioactivity and cytocompatibility of the carbon nanotubes and the polydopamine coated carbon nanotubes were assessed by immersion study in simulated body fluids and 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl tetrazolium bromide test using osteoblast cells (MC3T3-E1), respectively. As a result, it has been demonstrated that the introduction of polydopamine coating can greatly enhance the bioactivity and promote cell proliferation of the carbon nanotubes. The improvement of bioactive behavior is attributed to the good combination of catecholamines structure of the polydopamine and the structural advantages of carbon nanotubes as a framework material. It is anticipated that the polydopamine coated carbon nanotubes would find potential applications in bone tissue engineering and other biomedical areas.

The in vitro biomineralization and cytocompatibility of polydopamine coated carbon nanotubes

Energy Technology Data Exchange (ETDEWEB)

Yan Penghua [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, 18 Tianshui Middle Road, Lanzhou 730000 (China); Key Laboratory of Polymer Materials of Gansu Province, College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou 730070 (China); Wang Jinqing, E-mail: jqwang@licp.cas.cn [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, 18 Tianshui Middle Road, Lanzhou 730000 (China); Wang Lin; Liu Bin [School of Stomatology, Lanzhou University, Lanzhou 730000 (China); Lei Ziqiang [Key Laboratory of Polymer Materials of Gansu Province, College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou 730070 (China); Yang Shengrong, E-mail: sryang@lzb.ac.cn [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, 18 Tianshui Middle Road, Lanzhou 730000 (China); Key Laboratory of Polymer Materials of Gansu Province, College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou 730070 (China)

2011-03-15

In this work, polydopamine coated carbon nanotubes were firstly prepared by a simple and feasible route. Then, for comparison, the in vitro bioactivity and cytocompatibility of the carbon nanotubes and the polydopamine coated carbon nanotubes were assessed by immersion study in simulated body fluids and 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl tetrazolium bromide test using osteoblast cells (MC3T3-E1), respectively. As a result, it has been demonstrated that the introduction of polydopamine coating can greatly enhance the bioactivity and promote cell proliferation of the carbon nanotubes. The improvement of bioactive behavior is attributed to the good combination of catecholamines structure of the polydopamine and the structural advantages of carbon nanotubes as a framework material. It is anticipated that the polydopamine coated carbon nanotubes would find potential applications in bone tissue engineering and other biomedical areas.

In-vitro bioactivity, biocorrosion and antibacterial activity of silicon integrated hydroxyapatite/chitosan composite coating on 316 L stainless steel implants

Energy Technology Data Exchange (ETDEWEB)

Sutha, S.; Kavitha, K.; Karunakaran, G.; Rajendran, V., E-mail: veerajendran@gmail.com

2013-10-15

A simple and effective ultrasonication method was applied for the preparation of 0, 0.4, 0.8, 1.0 and 1.6 wt% silicon substituted hydroxyapatite (HAp) (SH). The Ca/P ratio of the synthesised SH nanoparticles were in the range of 1.58–1.70. Morphological changes were noticed in HAp with respect to the amount of Si from 0 to 1.6 wt%. The morphology of the particles changed from spherical shape to rod-like morphology with respect to the amount of Si which was confirmed using transmission electron microscopy. X-ray diffraction studies confirm the formation of phase pure SH nanoparticles without any secondary phase. Chitosan (CTS) blended SH nanocomposites coating on surgical grade 316 L stainless steel (316 L SS) implant was made by spin coating technique. The surface of the coated implant was characterised using scanning electron microscopy which confirms the uniform coating without cracks and pores. The increased corrosion resistance of the 1.6 wt% of SH/CTS-coated SS implant in the simulated body fluid (SBF) indicates the long-term biostability of SH composite-coated ceramics in vitro than the 0 wt% SH/CTS. The testing of SH/CTS nanocomposites with gram-positive and gram-negative bacterial strains confirms that the antibacterial ability improves with the higher substitution of Si. In addition, formation of bone-like apatite layer on the SH/CTS-coated implant in SBF was studied through SEM analysis and it confirms the ability to increase the HAp formation on the surface of 1.0 wt% SH/CTS-coated 316 L SS implant. Highlights: • Hydroxyapatite particles are prepared with various silicon concentration • Prepared composites are blended with chitosan and coated on the implant • Corrosion resistance in simulated body fluid improves its stability • Increase in silicon concentration improves the antibacterial activity • Coated plate exhibit high in-vitro bioactivity in simulated body fluid.

In-vitro bioactivity, biocorrosion and antibacterial activity of silicon integrated hydroxyapatite/chitosan composite coating on 316 L stainless steel implants

International Nuclear Information System (INIS)

Sutha, S.; Kavitha, K.; Karunakaran, G.; Rajendran, V.

2013-01-01

A simple and effective ultrasonication method was applied for the preparation of 0, 0.4, 0.8, 1.0 and 1.6 wt% silicon substituted hydroxyapatite (HAp) (SH). The Ca/P ratio of the synthesised SH nanoparticles were in the range of 1.58–1.70. Morphological changes were noticed in HAp with respect to the amount of Si from 0 to 1.6 wt%. The morphology of the particles changed from spherical shape to rod-like morphology with respect to the amount of Si which was confirmed using transmission electron microscopy. X-ray diffraction studies confirm the formation of phase pure SH nanoparticles without any secondary phase. Chitosan (CTS) blended SH nanocomposites coating on surgical grade 316 L stainless steel (316 L SS) implant was made by spin coating technique. The surface of the coated implant was characterised using scanning electron microscopy which confirms the uniform coating without cracks and pores. The increased corrosion resistance of the 1.6 wt% of SH/CTS-coated SS implant in the simulated body fluid (SBF) indicates the long-term biostability of SH composite-coated ceramics in vitro than the 0 wt% SH/CTS. The testing of SH/CTS nanocomposites with gram-positive and gram-negative bacterial strains confirms that the antibacterial ability improves with the higher substitution of Si. In addition, formation of bone-like apatite layer on the SH/CTS-coated implant in SBF was studied through SEM analysis and it confirms the ability to increase the HAp formation on the surface of 1.0 wt% SH/CTS-coated 316 L SS implant. Highlights: • Hydroxyapatite particles are prepared with various silicon concentration • Prepared composites are blended with chitosan and coated on the implant • Corrosion resistance in simulated body fluid improves its stability • Increase in silicon concentration improves the antibacterial activity • Coated plate exhibit high in-vitro bioactivity in simulated body fluid

Progresses in optimization strategy for radiolabeled molecular probes targeting integrin αvβ3

International Nuclear Information System (INIS)

Chen Haojun; Wu Hua

2012-01-01

Tumor angiogenesis is critical in the growth, invasion and metastasis of malignant tumors. The integrins, which express on many types of tumor cells and activated vascular endothelial cells, play an important role in regulation of the tumor angiogenesis. RGD peptide, which contains Arg-Gly-Asp sequence, binds specifically to integrin α v β 3 . Therefore, the radiolabeled RGD peptides may have broad application prospects in radionuclide imaging and therapy. Major research interests include the selection of radionuclides, modification and improvement of RGD structures. In this article, we give a review on research progresses in optimization strategy for radiolabeled molecular probes targeting integrin α v β 3 . (authors)

In vitro bioactivity and cytocompatibility of tricalcium silicate

Indian Academy of Sciences (India)

tricalcium silicate powder showed that it could induce bone- like apatite formation after ... ated by soaking them in SBF, cell adhesion and MTT assay, respectively. 2. .... tibility, which might be used as one of the bioactive coating materials and ...

Bioactivity and Surface Reactivity of RF-sputtered Calcium Phosphate Thin Films

NARCIS (Netherlands)

Wal, Edwin van der

2003-01-01

Calcium phosphates (CaP) are known to be bioactive, i.e. able to bond to bone. This makes CaPs very suitable to be aplied as thin coatings on bone-implants. In this work we studied the physicochemical behaviour of CaP coatings applied with radio frequency (RF) magnetron sputtering, a deposition

Endocytosis of Integrin-Binding Human Picornaviruses

Directory of Open Access Journals (Sweden)

Pirjo Merilahti

2012-01-01

Full Text Available Picornaviruses that infect humans form one of the largest virus groups with almost three hundred virus types. They include significant enteroviral pathogens such as rhino-, polio-, echo-, and coxsackieviruses and human parechoviruses that cause wide range of disease symptoms. Despite the economic importance of picornaviruses, there are no antivirals. More than ten cellular receptors are known to participate in picornavirus infection, but experimental evidence of their role in cellular infection has been shown for only about twenty picornavirus types. Three enterovirus types and one parechovirus have experimentally been shown to bind and use integrin receptors in cellular infection. These include coxsackievirus A9 (CV-A9, echovirus 9, and human parechovirus 1 that are among the most common and epidemic human picornaviruses and bind to αV-integrins via RGD motif that resides on virus capsid. In contrast, echovirus 1 (E-1 has no RGD and uses integrin α2β1 as cellular receptor. Endocytosis of CV-A9 has recently been shown to occur via a novel Arf6- and dynamin-dependent pathways, while, contrary to collagen binding, E-1 binds inactive β1 integrin and enters via macropinocytosis. In this paper, we review what is known about receptors and endocytosis of integrin-binding human picornaviruses.

The promotion of osseointegration of titanium surfaces by coating with silk protein sericin.

Science.gov (United States)

Nayak, Sunita; Dey, Tuli; Naskar, Deboki; Kundu, Subhas C

2013-04-01

A promising strategy to influence the osseointegration process around orthopaedic titanium implants is the immobilization of bioactive molecules. This recruits appropriate interaction between the surface and the tissue by directing cells adhesion, proliferation, differentiation and active matrix remodelling. In this study, we aimed to investigate the functionalization of metallic implant titanium with silk protein sericin. Titanium surface was immobilized with non-mulberry Antheraea mylitta sericin using glutaraldehyde as crosslinker. To analyse combinatorial effects the sericin immobilized titanium was further conjugated with integrin binding peptide sequence Arg-Gly-Asp (RGD) using ethyl (dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide as coupling agents. The surface of sericin immobilized titanium was characterized biophysically. Osteoblast-like cells were cultured on sericin and sericin/RGD functionalized titanium and found to be more viable than those on pristine titanium. The enhanced adhesion, proliferation, and differentiation of osteoblast cells were observed. RT-PCR analysis showed that mRNA expressions of bone sialoprotein, osteocalcin and alkaline phosphatase were upregulated in osteoblast cells cultured on sericin and sericin/RGD immobilized titanium substrates. Additionally, no significant amount of pro-inflammatory cytokines TNF-α, IL-1β and nitric oxide production were recorded when macrophages cells and osteoblast-macrophages co culture cells were grown on sericin immobilized titanium. The findings demonstrate that the sericin immobilized titanium surfaces are potentially useful bioactive coated materials for titanium-based medical implants. Copyright © 2013 Elsevier Ltd. All rights reserved.

Preparation of bioactive porous HA/PCL composite scaffolds

Energy Technology Data Exchange (ETDEWEB)

Zhao, J.; Guo, L.Y.; Yang, X.B. [Key Laboratory of Advanced Technologies of Materials (Ministry of Education), School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu 610031 (China); Weng, J. [Key Laboratory of Advanced Technologies of Materials (Ministry of Education), School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu 610031 (China)], E-mail: jweng@swjtu.cn

2008-12-30

Porous hydroxyapatite (HA) bioceramic scaffold has been widely attracted the attention to act as a three-dimensional (3D) template for cell adhesion, proliferation, differentiation and thus promoting bone and cartilage regeneration because of its osteoinduction. However, the porous bioceramic scaffold is fragile so that it is not suitable to be applied in clinic for bone repair or replacement. Therefore, it is significant to improve the mechanical property of porous HA bioceramics while the interconnected structure is maintained for tissue ingrowth in vivo. In the present research, a porous composite scaffold composed of HA scaffold and polycaprolactone (PCL) lining was fabricated by the method of polymer impregnating to produce HA scaffold coated with PCL lining. Subsequently, the composite scaffolds were deposited with biomimetic coating for improving the bioactivity. The HA/PCL composite scaffolds with improved mechanical property and bioactivity is expected to be a promising bone substitute in tissue engineering applications.

Preparation of bioactive porous HA/PCL composite scaffolds

International Nuclear Information System (INIS)

Zhao, J.; Guo, L.Y.; Yang, X.B.; Weng, J.

2008-01-01

Porous hydroxyapatite (HA) bioceramic scaffold has been widely attracted the attention to act as a three-dimensional (3D) template for cell adhesion, proliferation, differentiation and thus promoting bone and cartilage regeneration because of its osteoinduction. However, the porous bioceramic scaffold is fragile so that it is not suitable to be applied in clinic for bone repair or replacement. Therefore, it is significant to improve the mechanical property of porous HA bioceramics while the interconnected structure is maintained for tissue ingrowth in vivo. In the present research, a porous composite scaffold composed of HA scaffold and polycaprolactone (PCL) lining was fabricated by the method of polymer impregnating to produce HA scaffold coated with PCL lining. Subsequently, the composite scaffolds were deposited with biomimetic coating for improving the bioactivity. The HA/PCL composite scaffolds with improved mechanical property and bioactivity is expected to be a promising bone substitute in tissue engineering applications

Beyond the Matrix: The Many Non-ECM Ligands for Integrins

Directory of Open Access Journals (Sweden)

Bryce LaFoya

2018-02-01

Full Text Available The traditional view of integrins portrays these highly conserved cell surface receptors as mediators of cellular attachment to the extracellular matrix (ECM, and to a lesser degree, as coordinators of leukocyte adhesion to the endothelium. These canonical activities are indispensable; however, there is also a wide variety of integrin functions mediated by non-ECM ligands that transcend the traditional roles of integrins. Some of these unorthodox roles involve cell-cell interactions and are engaged to support immune functions such as leukocyte transmigration, recognition of opsonization factors, and stimulation of neutrophil extracellular traps. Other cell-cell interactions mediated by integrins include hematopoietic stem cell and tumor cell homing to target tissues. Integrins also serve as cell-surface receptors for various growth factors, hormones, and small molecules. Interestingly, integrins have also been exploited by a wide variety of organisms including viruses and bacteria to support infectious activities such as cellular adhesion and/or cellular internalization. Additionally, the disruption of integrin function through the use of soluble integrin ligands is a common strategy adopted by several parasites in order to inhibit blood clotting during hematophagy, or by venomous snakes to kill prey. In this review, we strive to go beyond the matrix and summarize non-ECM ligands that interact with integrins in order to highlight these non-traditional functions of integrins.

Structure, MC3T3-E1 cell response, and osseointegration of macroporous titanium implants covered by a bioactive microarc oxidation coating with microporous structure.

Science.gov (United States)

Zhou, Rui; Wei, Daqing; Cheng, Su; Feng, Wei; Du, Qing; Yang, Haoyue; Li, Baoqiang; Wang, Yaming; Jia, Dechang; Zhou, Yu

2014-04-09

Macroporous Ti with macropores of 50-400 μm size is prepared by sintering Ti microbeads with different diameters of 100, 200, 400, and 600 μm. Bioactive microarc oxidation (MAO) coatings with micropores of 2-5 μm size are prepared on the macroporous Ti. The MAO coatings are composed of a few TiO2 nanocrystals and lots of amorphous phases with Si, Ca, Ti, Na, and O elements. Compared to compact Ti, the MC3T3-E1 cell attachment is prolonged on macroporous Ti without and with MAO coatings; however, the cell proliferation number increases. These results are contributed to the effects of the space structure of macroporous Ti and the surface chemical feature and element dissolution of the MAO coatings during the cell culture. Macroporous Ti both without and with MAO coatings does not cause any adverse effects in vivo. The new bone grows well into the macropores and micropores of macroporous Ti with MAO coatings, showing good mechanical properties in vivo compared to Ti, MAO-treated Ti, and macroporous Ti because of its excellent osseointegration. Moreover, the MAO coatings not only show a high interface bonding strength with new bones but also connect well with macroporous Ti. Furthermore, the pushing out force for macroporous Ti with MAO coatings increases significantly with increasing microbead diameter.

In vitro study of nanostructured diopside coating on Mg alloy orthopedic implants

International Nuclear Information System (INIS)

Razavi, Mehdi; Fathi, Mohammadhossein; Savabi, Omid; Vashaee, Daryoosh; Tayebi, Lobat

2014-01-01

The high corrosion rate of Mg alloys has hindered their application in various areas, particularly for orthopedic applications. In order to decrease the corrosion rate and to improve the bioactivity, mechanical stability and cytocompatibility of the Mg alloy, nanostructured diopside (CaMgSi 2 O 6 ) has been coated on AZ91 Mg alloy using a combined micro arc oxidation (MAO) and electrophoretic deposition (EPD) method. The crystalline structure, the morphology and the composition of the samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Electrochemical corrosion test, immersion test, and compression test were used to evaluate the corrosion resistance, the in vitro bioactivity and the mechanical stability of the samples, respectively. The cytocompatibility of the samples was tested by the cell viability and the cell attachment of L-929 cells. The results confirmed that the diopside coating not only slows down the corrosion rate, but also enhances the in vitro bioactivity, mechanical stability and cytocompatibility of AZ91 Mg alloy. Therefore, Mg alloy coated with nanostructured diopside offers a promising approach for biodegradable bone implants. - Highlights: • The diopside coating was applied on Mg alloy using the combined MAO and EPD methods. • The corrosion resistance of the diopside coated Mg alloy was noticeably improved. • The in vitro bioactivity of the diopside coated Mg alloy was considerably increased. • The mechanical stability of biodegradable Mg alloy was enhanced by diopside coating. • The cytocompatibility of the Mg alloy was improved employing diopside coating

In vitro study of nanostructured diopside coating on Mg alloy orthopedic implants

Energy Technology Data Exchange (ETDEWEB)

Razavi, Mehdi, E-mail: mehdi.razavi@okstate.edu [Biomaterials Research Group, Department of Materials Engineering, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of); Torabinejad Dental Research Center, School of Dentistry, Isfahan University of Medical Sciences, Isfahan 81746-73461 (Iran, Islamic Republic of); School of Materials Science and Engineering, Helmerich Advanced Technology Research Center, Oklahoma State University, Tulsa, OK 74106 (United States); School of Electrical and Computer Engineering, Helmerich Advanced Technology Research Center, Oklahoma State University, Tulsa, OK 74106 (United States); Fathi, Mohammadhossein [Biomaterials Research Group, Department of Materials Engineering, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of); Dental Materials Research Center, Isfahan University of Medical Sciences, Isfahan (Iran, Islamic Republic of); Savabi, Omid [Torabinejad Dental Research Center, School of Dentistry, Isfahan University of Medical Sciences, Isfahan 81746-73461 (Iran, Islamic Republic of); Vashaee, Daryoosh [School of Electrical and Computer Engineering, Helmerich Advanced Technology Research Center, Oklahoma State University, Tulsa, OK 74106 (United States); Tayebi, Lobat, E-mail: lobat.tayebi@okstate.edu [School of Materials Science and Engineering, Helmerich Advanced Technology Research Center, Oklahoma State University, Tulsa, OK 74106 (United States); School of Chemical Engineering, Oklahoma State University, Stillwater, OK 74078 (United States)

2014-08-01

The high corrosion rate of Mg alloys has hindered their application in various areas, particularly for orthopedic applications. In order to decrease the corrosion rate and to improve the bioactivity, mechanical stability and cytocompatibility of the Mg alloy, nanostructured diopside (CaMgSi{sub 2}O{sub 6}) has been coated on AZ91 Mg alloy using a combined micro arc oxidation (MAO) and electrophoretic deposition (EPD) method. The crystalline structure, the morphology and the composition of the samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Electrochemical corrosion test, immersion test, and compression test were used to evaluate the corrosion resistance, the in vitro bioactivity and the mechanical stability of the samples, respectively. The cytocompatibility of the samples was tested by the cell viability and the cell attachment of L-929 cells. The results confirmed that the diopside coating not only slows down the corrosion rate, but also enhances the in vitro bioactivity, mechanical stability and cytocompatibility of AZ91 Mg alloy. Therefore, Mg alloy coated with nanostructured diopside offers a promising approach for biodegradable bone implants. - Highlights: • The diopside coating was applied on Mg alloy using the combined MAO and EPD methods. • The corrosion resistance of the diopside coated Mg alloy was noticeably improved. • The in vitro bioactivity of the diopside coated Mg alloy was considerably increased. • The mechanical stability of biodegradable Mg alloy was enhanced by diopside coating. • The cytocompatibility of the Mg alloy was improved employing diopside coating.

Multifunctional bioactive and improving the performance durability nanocoatings for finishing PET/CO woven fabrics by the sol–gel method

Energy Technology Data Exchange (ETDEWEB)

Kowalczyk, Dorota, E-mail: dkowalczyk@iw.lodz.pl; Brzeziński, Stefan; Kamińska, Irena

2015-11-15

The paper presents the results of studies on multifunctional thin-coatings of textiles, simultaneously imparting to them bioactive properties in relations to bacteria and fungi as well as an increased abrasion resistance and anti-pilling effect with the use of modified hybrid materials produced by the sol–gel method from two precursors: (3-glycidoxypropyl)trimethoxysilane (GPTMS) and aluminum isopropoxide (ALIPO). The sol obtained was modified with bioactive particles in the form of nanopowder of metallic silver and copper alloy (Ag/Cu). Al{sub 2}O{sub 3}/SiO{sub 2} sol containing nanoparticles of Ag/Cu alloy was deposited on a polyester/cotton blend woven fabric (PET/CO 67/33) by the padding method. After drying and curing process, a thin and elastic bioactive silica coating was obtained on the fabric fibers surfaces. The fabrics with deposited nanocoatings were characterized by very good bioactive properties and increased resistance to abrasion and formation of pilling. - Highlights: • Multifunctional thin coating layer was prepared on the fabric using sol–gel method. • Modification of the hybrid Al{sub 2}O{sub 3}/SiO{sub 2} sol by Ag/Cu alloy nanoparticles. • Bioactive fabric created by deposition of Al{sub 2}O{sub 3}/SiO{sub 2} sol with Ag/Cu. • 30% increase the abrasion resistance of the thin coating fabric.

Ligand-Occupied Integrin Internalization Links Nutrient Signaling to Invasive Migration

Directory of Open Access Journals (Sweden)

Elena Rainero

2015-01-01

Full Text Available Integrin trafficking is key to cell migration, but little is known about the spatiotemporal organization of integrin endocytosis. Here, we show that α5β1 integrin undergoes tensin-dependent centripetal movement from the cell periphery to populate adhesions located under the nucleus. From here, ligand-engaged α5β1 integrins are internalized under control of the Arf subfamily GTPase, Arf4, and are trafficked to nearby late endosomes/lysosomes. Suppression of centripetal movement or Arf4-dependent endocytosis disrupts flow of ligand-bound integrins to late endosomes/lysosomes and their degradation within this compartment. Arf4-dependent integrin internalization is required for proper lysosome positioning and for recruitment and activation of mTOR at this cellular subcompartment. Furthermore, nutrient depletion promotes subnuclear accumulation and endocytosis of ligand-engaged α5β1 integrins via inhibition of mTORC1. This two-way regulatory interaction between mTORC1 and integrin trafficking in combination with data describing a role for tensin in invasive cell migration indicate interesting links between nutrient signaling and metastasis.

Noninvasive imaging of tumor integrin expression using 18F-labeled RGD dimer peptide with PEG4 linkers

International Nuclear Information System (INIS)

Liu, Zhaofei; Liu, Shuanglong; Wang, Fan; Liu, Shuang; Chen, Xiaoyuan

2009-01-01

Various radiolabeled Arg-Gly-Asp (RGD) peptides have been previously investigated for tumor integrin α v β 3 imaging. To further develop RGD radiotracers with enhanced tumor-targeting efficacy and improved in vivo pharmacokinetics, we designed a new RGD homodimeric peptide with two PEG 4 spacers (PEG 4 = 15-amino-4,7,10,13-tetraoxapentadecanoic acid) between the two monomeric RGD motifs and one PEG 4 linker on the glutamate α-amino group ( 18 F-labeled PEG 4 -E[PEG 4 -c(RGDfK)] 2 , P-PRGD2), as a promising agent for noninvasive imaging of integrin expression in mouse models. P-PRGD2 was labeled with 18 F via 4-nitrophenyl 2- 18 F-fluoropropionate ( 18 F-FP) prosthetic group. In vitro and in vivo characteristics of the new dimeric RGD peptide tracer 18 F-FP-P-PRGD2 were investigated and compared with those of 18 F-FP-P-RGD2 ( 18 F-labeled RGD dimer without two PEG 4 spacers between the two RGD motifs). The ability of 18 F-FP-P-PRGD2 to image tumor vascular integrin expression was evaluated in a 4T1 murine breast tumor model. With the insertion of two PEG 4 spacers between the two RGD motifs, 18 F-FP-P-PRGD2 showed enhanced integrin α v β 3 -binding affinity, increased tumor uptake and tumor-to-nontumor background ratios compared with 18 F-FP-P-RGD2 in U87MG tumors. MicroPET imaging with 18 F-FP-P-PRGD2 revealed high tumor contrast and low background in tumor-bearing nude mice. Biodistribution studies confirmed the in vivo integrin α v β 3 -binding specificity of 18 F-FP-P-RGD2. 18 F-FP-P-PRGD2 can specifically image integrin α v β 3 on the activated endothelial cells of tumor neovasculature. 18 F-FP-P-PRGD2 can provide important information on integrin expression on the tumor vasculature. The high integrin binding affinity and specificity, excellent pharmacokinetic properties and metabolic stability make the new RGD dimeric tracer 18 F-FP-P-PRGD2 a promising agent for PET imaging of tumor angiogenesis and for monitoring the efficacy of antiangiogenic

Plasma spraying of bioactive glass-ceramics containing bovine bone

Directory of Open Access Journals (Sweden)

Annamária Dobrádi

2017-06-01

Full Text Available Natural bone derived glass-ceramics are promising biomaterials for implants. However, due to their price and weak mechanical properties they are preferably applied as coatings on load bearing implants. This paper describes result obtained by plasma spraying of bioactive glass-ceramics containing natural bone onto selected implant materials, such as stainless steel, alumina, and titanium alloy. Adhesion of plasma sprayed coating was tested by computed X-ray tomography and SEM of cross sections. The results showed defect free interface between the coating and substrate, without cracks or gaps. Dissolution rate of the coating in simulated body fluid (SBF was readily controlled by the bone additives (phase composition, as well as microstructure. The SBF treatment of the plasma sprayed coating did not influence the boundary between the coating and substrate.

Pectin-honey coating as novel dehydrating bioactive agent for cut fruit: Enhancement of the functional properties of coated dried fruits.

Science.gov (United States)

Santagata, Gabriella; Mallardo, Salvatore; Fasulo, Gabriella; Lavermicocca, Paola; Valerio, Francesca; Di Biase, Mariaelena; Di Stasio, Michele; Malinconico, Mario; Volpe, Maria Grazia

2018-08-30

In this paper, a novel and sustainable process for the fruit dehydration was described. Specifically, edible coatings based on pectin and honey were prepared and used as dehydrating and antimicrobial agents of cut fruit samples, in this way promoting the fruit preservation from irreversible deteriorative processes. Pectin-honey coating was tested on apple, cantaloupe melon, mango and pineapple. The analysis were performed also on uncoated dehydrated fruits (control). The coated fruit evidenced enhanced dehydration percentage, enriched polyphenol and vitamin C contents, improved antioxidant activity and volatile molecules profile. Moreover, the antimicrobial activity against Pseudomonas and Escherichia coli was assessed. Finally, morphological analysis performed on fruit fractured surface, highlighted the formation of a non-sticky and homogeneous thin layer. These outcomes suggested that the novel fruit dehydration process, performed by using pectin-honey coating, was able to both preserve the safety and quality of dehydrated fruits, and enhance their authenticity and naturalness. Copyright © 2018 Elsevier Ltd. All rights reserved.

Laminin-binding integrins and their tetraspanin partners as potential antimetastatic targets

Science.gov (United States)

Stipp, Christopher S.

2010-01-01

Within the integrin family of cell adhesion receptors, integrins α3β1, α6β1, α6β4 and α7β1 make up a laminin-binding subfamily. The literature is divided on the role of these laminin-binding integrins in metastasis, with different studies indicating either pro- or antimetastatic functions. The opposing roles of the laminin-binding integrins in different settings might derive in part from their unusually robust associations with tetraspanin proteins. Tetraspanins organise integrins into multiprotein complexes within discrete plasma membrane domains termed tetraspanin-enriched microdomains (TEMs). TEM association is crucial to the strikingly rapid cell migration mediated by some of the laminin-binding integrins. However, emerging data suggest that laminin-binding integrins also promote the stability of E-cadherin-based cell–cell junctions, and that tetraspanins are essential for this function as well. Thus, TEM association endows the laminin-binding integrins with both pro-invasive functions (rapid migration) and anti-invasive functions (stable cell junctions), and the composition of TEMs in different cell types might help determine the balance between these opposing activities. Unravelling the tetraspanin control mechanisms that regulate laminin-binding integrins will help to define the settings where inhibiting the function of these integrins would be helpful rather than harmful, and may create opportunities to modulate integrin activity in more sophisticated ways than simple functional blockade. PMID:20078909

Electrophoretic deposition of mesoporous bioactive glass on glass-ceramic foam scaffolds for bone tissue engineering.

Science.gov (United States)

Fiorilli, Sonia; Baino, Francesco; Cauda, Valentina; Crepaldi, Marco; Vitale-Brovarone, Chiara; Demarchi, Danilo; Onida, Barbara

2015-01-01

In this work, the coating of 3-D foam-like glass-ceramic scaffolds with a bioactive mesoporous glass (MBG) was investigated. The starting scaffolds, based on a non-commercial silicate glass, were fabricated by the polymer sponge replica technique followed by sintering; then, electrophoretic deposition (EPD) was applied to deposit a MBG layer on the scaffold struts. EPD was also compared with other techniques (dipping and direct in situ gelation) and it was shown to lead to the most promising results. The scaffold pore structure was maintained after the MBG coating by EPD, as assessed by SEM and micro-CT. In vitro bioactivity of the scaffolds was assessed by immersion in simulated body fluid and subsequent evaluation of hydroxyapatite (HA) formation. The deposition of a MBG coating can be a smart strategy to impart bioactive properties to the scaffold, allowing the formation of nano-structured HA agglomerates within 48 h from immersion, which does not occur on uncoated scaffold surfaces. The mechanical properties of the scaffold do not vary after the EPD (compressive strength ~19 MPa, fracture energy ~1.2 × 10(6) J m(-3)) and suggest the suitability of the prepared highly bioactive constructs as bone tissue engineering implants for load-bearing applications.

Integrin-based meningioma cell migration is promoted by photon but not by carbon-ion irradiation

International Nuclear Information System (INIS)

Simon, Florian; Dittmar, Jan-Oliver; Orschiedt, Lena; Weber, Klaus-Josef; Debus, Juergen; Rieken, Stefan; Brons, Stephan; Urbschat, Steffi; Combs, Stephanie E.

2015-01-01

Sublethal doses of photon irradiation (IR) are suspected to increase tumor cell migration and support locoregional recurrence of disease, which has already been shown in other cell lines. This manuscript describes the effect of photon and carbon-ion IR on WHO class I meningioma cell migration and provides an approach to the underlying cellular mechanisms. Meningioma cells were gained operatively at the university hospital in Homburg/Saar, Germany. For migration, membranes (8-μm pore sizes) were coated with collagen I, with collagen IV, and with fibronectin. Cells were analyzed in migration experiments with or without serum stimulation, with or without photon and carbon IR 24 h prior to experiments, and with or without integrin antibodies. Fluorescence-activated cell sorting (FACS) analyses of the integrins ανβ 1 , ανβ 3 , and ανβ 5 were performed without IR and 6, 12 and 24 h after IR. Enzyme-linked immunosorbent assay (ELISA) analyses of matrix metalloproteinases (MMP)-2 and MMP-9 were realized with and without IR after cells were cultured on collagen I, collagen IV, or fibronectin for 24 h. Cells and supernatants for FACS and ELISA were stored at - 18 C. The significance level was set at 5 % using both Student's t test and two-way ANOVA. Migration of meningioma cells was serum-inducible (p < 0.001). It could be increased by photon IR (p < 0.02). The integrins ανβ 1 and ανβ 5 showed a 21 and 11 % higher expression after serum stimulation (not significant), respectively, and ανβ 1 expression was raised by 14 % (p = 0.0057) after photon IR. Antibody blockage of the integrins ανβ 1 and ανβ 5 inhibited serum- and photon-induced migration. Expression of MMP-2 and MMP-9 remained unchanged after both IR and fetal bovine serum (FBS). Carbon-ion IR left both integrin expression and meningioma cell migration unaffected. Photon but not carbon-ion IR promotes serum-based meningioma cell migration. Fibronectin receptor integrin ανβ 1 signaling

Breast Cancer Cells in Three-dimensional Culture Display an Enhanced Radioresponse after Coordinate Targeting of Integrin ?5?1 and Fibronectin

Energy Technology Data Exchange (ETDEWEB)

Nam, Jin-Min; Onodera, Yasuhito; Bissell, Mina J; Park, Catherine C

2010-04-07

Tactics to selectively enhance cancer radioresponse are of great interest. Cancer cells actively elaborate and remodel their extracellular matrix (ECM) to aid in survival and progression. Previous work has shown that {beta}1-integrin inhibitory antibodies can enhance the growth-inhibitory and apoptotic responses of human breast cancer cell lines to ionizing radiation, either when cells are cultured in three-dimensional laminin-rich ECM (3D lrECM) or grown as xenografts in mice. Here, we show that a specific {alpha} heterodimer of {beta}1-integrin preferentially mediates a prosurvival signal in human breast cancer cells that can be specifically targeted for therapy. 3D lrECM culture conditions were used to compare {alpha}-integrin heterodimer expression in malignant and nonmalignant cell lines. Under these conditions, we found that expression of {alpha}5{beta}1-integrin was upregulated in malignant cells compared with nonmalignant breast cells. Similarly, we found that normal and oncofetal splice variants of fibronectin, the primary ECM ligand of {alpha}5{beta}1-integrin, were also strikingly upregulated in malignant cell lines compared with nonmalignant acini. Cell treatment with a peptide that disrupts the interactions of {alpha}5{beta}1-integrin with fibronectin promoted apoptosis in malignant cells and further heightened the apoptotic effects of radiation. In support of these results, an analysis of gene expression array data from breast cancer patients revealed an association of high levels of {alpha}5-integrin expression with decreased survival. Our findings offer preclinical validation of fibronectin and {alpha}5{beta}1-integrin as targets for breast cancer therapy.

Deoxycholic acid promotes development of gastroesophageal reflux disease and Barrett's oesophagus by modulating integrin-αv trafficking.

Science.gov (United States)

Prichard, David O; Byrne, Anne Marie; Murphy, James O; Reynolds, John V; O'Sullivan, Jacintha; Feighery, Ronan; Doyle, Brendan; Eldin, Osama Sharaf; Finn, Stephen P; Maguire, Aoife; Duff, Deirdre; Kelleher, Dermot P; Long, Aideen

2017-12-01

The fundamental mechanisms underlying erosive oesophagitis and subsequent development of Barrett's oesophagus (BO) are poorly understood. Here, we investigated the contribution of specific components of the gastric refluxate on adhesion molecules involved in epithelial barrier maintenance. Cell line models of squamous epithelium (HET-1A) and BO (QH) were used to examine the effects of bile acids on cell adhesion to extracellular matrix proteins (Collagen, laminin, vitronectin, fibronectin) and expression of integrin ligands (α 3 , α 4, α 5 , α 6 and α ν ). Experimental findings were validated in human explant oesophageal biopsies, a rat model of gastroesophageal reflux disease (GORD) and in patient tissue microarrays. The bile acid deoxycholic acid (DCA) specifically reduced adhesion of HET-1A cells to vitronectin and reduced cell-surface expression of integrin-α ν via effects on endocytic recycling processes. Increased expression of integrin-α v was observed in ulcerated tissue in a rat model of GORD and in oesophagitis and Barrett's intestinal metaplasia patient tissue compared to normal squamous epithelium. Increased expression of integrin-α ν was observed in QH BO cells compared to HET-1A cells. QH cells were resistant to DCA-mediated loss of adhesion and reduction in cell-surface expression of integrin-α ν . We demonstrated that a specific component of the gastric refluxate, DCA, affects the epithelial barrier through modulation of integrin α ν expression, providing a novel mechanism for bile acid-mediated erosion of oesophageal squamous epithelium and promotion of BO. Strategies aimed at preventing bile acid-mediated erosion should be considered in the clinical management of patients with GORD. © 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Bioactivity and electrochemical behavior of hydroxyapatite-silicon-multi walled carbon nano-tubes composite coatings synthesized by EPD on NiTi alloys in simulated body fluid

Energy Technology Data Exchange (ETDEWEB)

Khalili, V., E-mail: V_khalili@sut.ac.ir [Department of Materials Engineering, Engineering Faculty, University of Bonab, Bonab (Iran, Islamic Republic of); Khalil-Allafi, J. [Research Center for Advanced Materials and Mineral Processing, Faculty of Materials Engineering, Sahand University of Technology, Tabriz (Iran, Islamic Republic of); Frenzel, J.; Eggeler, G. [Institute for Materials, Faculty of Mechanical Engineering, Ruhr-University Bochum, 44801 Bochum (Germany)

2017-02-01

In order to improve the surface bioactivity of NiTi bone implant and corrosion resistance, hydroxyapatite coating with addition of 20 wt% silicon, 1 wt% multi walled carbon nano-tubes and both of them were deposited on a NiTi substrate using a cathodic electrophoretic method. The apatite formation ability was estimated using immersion test in the simulated body fluid for 10 days. The SEM images of the surface of coatings after immersion in simulated body fluid show that the presence of silicon in the hydroxyapatite coatings accelerates in vitro growth of apatite layer on the coatings. The Open-circuit potential and electrochemical impedance spectroscopy were measured to evaluate the electrochemical behavior of the coatings in the simulated body fluid at 37 °C. The results indicate that the compact structure of hydroxyapatite-20 wt% silicon and hydroxyapatite-20 wt% silicon-1 wt% multi walled carbon nano-tubes coatings could efficiently increase the corrosion resistance of NiTi substrate. - Highlights: • The composite coatings of HA, Si and MWCNTs was prepared using electrophoretic deposition. • The presence of 1 wt.% MWCNTs in the HA coating provides more nucleation cites of apatite crystallites in SBF. • The presence of Si in HA coating increases the growth rate of apatite crystallites with the Ca/P atomic ratio of 1.67. • The EIS indicate the compact HA-20%Si and HA-20%Si-1%MWCNTs coatings efficiently increase corrosion resistance of NiTi. • The porous HA and HA-1%MWCNTs do not increase significantly corrosion resistance due to the easy diffusion path.

Chemical changes in DMP1-null murine bone & silica based pecvd coatings for titanium implant osseoapplications

Science.gov (United States)

Maginot, Megen

In order to improve clinical outcomes in bone-implant systems, a thorough understanding of both local bone chemistry and implant surface chemistry is necessary. This study consists, therefore, of two main parts: one focused on determining the nature of the changes in bone chemistry in a DMP1-null transgenic disease model and the other on the development of amorphous silica-based coatings for potential use as titanium bone implant coatings. For the study of bone mineral in the DMP1 transgenic model, which is known to have low serum phosphate levels, transgenic DMP1-null and wild type mice were fed a high phosphate diet, sacrificed, and had their long bone harvested. This bone was characterized using SEM, FTIR, microCT and XANES and compared to DMP1-null and wild type control groups to assess the therapeutic effect of high Pi levels on the phenotype and the role of DMP1 in mineralization in vivo. Findings suggest that though the high phosphate diet results in restoring serum phosphate levels, it does not completely rescue the bone mineral phenotype at an ultrastructural level and implicates DMP1 in phosphate nucleation. Since plasma enhanced chemical vapor deposition (PECVD) silica like coatings have not previously been fabricated for use in oessoapplications, the second part of this study initially focused on the characterization of novel SiOx chemistries fabricated via a chemical vapor deposition process that were designed specifically to act as bioactive coatings with a loose, hydrogenated structure. These coatings were then investigated for their potential initial stage response to bone tissue through immersion in a simulated body fluid and through the culture of MC3T3 cells on the coating surfaces. Coating surfaces were characterized by SEM, FTIR, contact angle measurements, and XANES. Coating dissolution and ionic release were also investigated by ICP-OES. Findings suggest that some SiOx chemistries may form a bioactive coating while more highly substituted

Biomimetic surface coatings from modular amphiphilic proteins

Science.gov (United States)

Harden, James; Wan, Fan; Fischer, Stephen; Dick, Scott

2010-03-01

Recombinant DNA methods have been used to develop a library of diblock protein polymers for creating designer biofunctional interfaces. These proteins are composed of a surface-active, amphiphilic block joined to a disordered, water soluble block with an end terminal bioactive domain. The amphiphilic block has a strong affinity for many synthetic polymer surfaces, providing a facile means of imparting biological functionality to otherwise bio-neutral materials through physical self-assembly. We have incorporated a series of bioactive end domains into this diblock motif, including sequences that encode specific cell binding and signaling functions of extracellular matrix constituents (e.g. RGD and YIGSR). In this talk, we show that these diblock constructs self-assemble into biofunctional surface coatings on several model synthetic polymer materials. We demonstrate that surface adsorption of the proteins has minimal impacts on the presentation of the bioactive domains in the soluble block, and through the use of microscopic and cell proliferation assays, we show that the resulting biofunctional interfaces are capable of inducing appropriate cellular responses in a variety of human cell types.

Gfi1b controls integrin signaling-dependent cytoskeleton dynamics and organization in megakaryocytes.

Science.gov (United States)

Beauchemin, Hugues; Shooshtarizadeh, Peiman; Vadnais, Charles; Vassen, Lothar; Pastore, Yves D; Möröy, Tarik

2017-03-01

Mutations in GFI1B are associated with inherited bleeding disorders called GFI1B -related thrombocytopenias. We show here that mice with a megakaryocyte-specific Gfi1b deletion exhibit a macrothrombocytopenic phenotype along a megakaryocytic dysplasia reminiscent of GFI1B -related thrombocytopenia. GFI1B deficiency increases megakaryocyte proliferation and affects their ploidy, but also abrogates their responsiveness towards integrin signaling and their ability to spread and reorganize their cytoskeleton. Gfi1b -null megakaryocytes are also unable to form proplatelets, a process independent of integrin signaling. GFI1B-deficient megakaryocytes exhibit aberrant expression of several components of both the actin and microtubule cytoskeleton, with a dramatic reduction of α-tubulin. Inhibition of FAK or ROCK, both important for actin cytoskeleton organization and integrin signaling, only partially restored their response to integrin ligands, but the inhibition of PAK, a regulator of the actin cytoskeleton, completely rescued the responsiveness of Gfi1b -null megakaryocytes to ligands, but not their ability to form proplatelets. We conclude that Gfi1b controls major functions of megakaryocytes such as integrin-dependent cytoskeleton organization, spreading and migration through the regulation of PAK activity whereas the proplatelet formation defect in GFI1B-deficient megakaryocytes is due, at least partially, to an insufficient α-tubulin content. Copyright© Ferrata Storti Foundation.

Investigation of calcium phosphate coatings for biomedical applications

International Nuclear Information System (INIS)

Yusof Abdullah; Idris Besar; Muhammad Jamal Md Isa; Mohamad Abd Razak; Hyzan Mohd Yusof

1999-01-01

Calcium phosphate is the main constituent of our bone and tooth minerals. The use of this bioactive material for coating implant such as artificial joint prosthesis, therefore, can promote biological fixation and enhance biocompatibility. Our initial work has been focused on the evaluation of experimental conditions of coating preparation and the effects of post-deposition calcium phosphate coatings on stainless steel substrates. The coating layers were produced by the precipitation technique and coatings were carried out in sol-gel by the dipping method. For comparison purposes a wet method was used to obtain a fine calcium phosphate ceramic powder for fabrication of microcrystal suspension used as a coating material. Scanning electron microscopy (SEM), energy dispersive microanalysis (EDS), energy dispersive x-ray fluorescence (EDXRF) and x-ray diffraction (XRD) were used to characterise the morphology, chemical composition and structure of the coatings. The results showed that the dip coating of stainless steel substrates using viscous solutions lead to the formation of porous calcium phosphate layers. These results suggested that fabrication of bioactive calcium phosphate coatings using this route offers significant advantages over the currently used methods due to considerably lower temperature process involved and may produce better result for substrates with complex shapes

Epitopes in α8β1 and other RGD-binding integrins delineate classes of integrin-blocking antibodies and major binding loops in α subunits

OpenAIRE

Norihisa Nishimichi; Nagako Kawashima; Yasuyuki Yokosaki

2015-01-01

Identification of epitopes for integrin-blocking monoclonal antibodies (mAbs) has aided our understanding of structure-function relationship of integrins. We mapped epitopes of chicken anti-integrin-α8-subunit-blocking mAbs by mutational analyses, examining regions that harboured all mapped epitopes recognized by mAbs against other α-subunits in the RGD-binding-integrin subfamily. Six mAbs exhibited blocking function, and these mAbs recognized residues on the same W2:41-loop on the top-face o...

Human Parechovirus 1 Infection Occurs via αVβ1 Integrin.

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Pirjo Merilahti

Full Text Available Human parechovirus 1 (HPeV-1 (family Picornaviridae is a global cause of pediatric respiratory and CNS infections for which there is no treatment. Although biochemical and in vitro studies have suggested that HPeV-1 binds to αVβ1, αVβ3 and αVβ6 integrin receptor(s, the actual cellular receptors required for infectious entry of HPeV-1 remain unknown. In this paper we analyzed the expression profiles of αVβ1, αVβ3, αVβ6 and α5β1 in susceptible cell lines (A549, HeLa and SW480 to identify which integrin receptors support HPeV-1 internalization and/or replication cycle. We demonstrate by antibody blocking assay, immunofluorescence microscopy and RT-qPCR that HPeV-1 internalizes and replicates in cell lines that express αVβ1 integrin but not αVβ3 or αVβ6 integrins. To further study the role of β1 integrin, we used a mouse cell line, GE11-KO, which is deficient in β1 expression, and its derivate GE11-β1 in which human integrin β1 subunit is overexpressed. HPeV-1 (Harris strain and three clinical HPeV-1 isolates did not internalize into GE11-KO whereas GE11-β1 supported the internalization process. An integrin β1-activating antibody, TS2/16, enhanced HPeV-1 infectivity, but infection occurred in the absence of visible receptor clustering. HPeV-1 also co-localized with β1 integrin on the cell surface, and HPeV-1 and β1 integrin co-endocytosed into the cells. In conclusion, our results demonstrate that in some cell lines the cellular entry of HPeV-1 is primarily mediated by the active form of αVβ1 integrin without visible receptor clustering.

Characterization of a biomimetic coating on dense and porous titanium substrates

Energy Technology Data Exchange (ETDEWEB)

Rocha, M.N. da; Pereira, L.C. [Coordenacao dos Programas de Pos-Graduacao de Engenharia (PEMM/COPPE/UFRJ), RJ (Brazil). Programa de Engenharia Metalurgica e de Materiais; Ribeiro, A.A.; Oliveira, M.V. de, E-mail: marize.varella@int.gov.b [Instituto Nacional de Tecnologia (INT), Rio de Janeiro, RJ (Brazil); Andrade, M.C. de [Universidade do Estado do Rio de Janeiro (IPRJ/UERJ), Nova Friburgo, RJ (Brazil). Inst. Politecnico

2010-07-01

Bioactive materials have been studied as coatings on bioinert subtracts. Thus, it is possible to combine the bioactivity of materials such as calcium phosphate with the excellent mechanical properties of metals. Titanium (Ti) implants can be bioactivated by a biomimetic precipitation method. This study introduces a biomimetic method under a simplified solution (SS) with calcium and phosphorus ions. As substrates, commercially pure Ti sheet and micro-porous Ti samples produced by powder metallurgy were used. The substrates were submitted to chemical and heat treating and then immersed in the SS for 7, 14, 21 days. Surface roughness was evaluated by confocal scanning optical microscopy. Coating characterization was performed by scanning electron microscopy and high resolution X-ray diffraction (XRD). The results showed calcium phosphate crystal morphologies observed in all samples, which was confirmed by XRD phase identifications. These results reveal the solution potential for coating Ti substrates. (author)

Characterization of a biomimetic coating on dense and porous titanium substrates

International Nuclear Information System (INIS)

Rocha, M.N. da; Pereira, L.C.; Andrade, M.C. de

2010-01-01

Bioactive materials have been studied as coatings on bioinert subtracts. Thus, it is possible to combine the bioactivity of materials such as calcium phosphate with the excellent mechanical properties of metals. Titanium (Ti) implants can be bioactivated by a biomimetic precipitation method. This study introduces a biomimetic method under a simplified solution (SS) with calcium and phosphorus ions. As substrates, commercially pure Ti sheet and micro-porous Ti samples produced by powder metallurgy were used. The substrates were submitted to chemical and heat treating and then immersed in the SS for 7, 14, 21 days. Surface roughness was evaluated by confocal scanning optical microscopy. Coating characterization was performed by scanning electron microscopy and high resolution X-ray diffraction (XRD). The results showed calcium phosphate crystal morphologies observed in all samples, which was confirmed by XRD phase identifications. These results reveal the solution potential for coating Ti substrates. (author)

Silk-Silk Interactions between Silkworm Fibroin and Recombinant Spider Silk Fusion Proteins Enable the Construction of Bioactive Materials.

Science.gov (United States)

Nilebäck, Linnea; Chouhan, Dimple; Jansson, Ronnie; Widhe, Mona; Mandal, Biman B; Hedhammar, My

2017-09-20

Natural silk is easily accessible from silkworms and can be processed into different formats suitable as biomaterials and cell culture matrixes. Recombinant DNA technology enables chemical-free functionalization of partial silk proteins through fusion with peptide motifs and protein domains, but this constitutes a less cost-effective production process. Herein, we show that natural silk fibroin (SF) can be used as a bulk material that can be top-coated with a thin layer of the recombinant spider silk protein 4RepCT in fusion with various bioactive motifs and domains. The coating process is based on a silk assembly to achieve stable interactions between the silk types under mild buffer conditions. The assembly process was studied in real time by quartz crystal microbalance with dissipation. Coatings, electrospun mats, and microporous scaffolds were constructed from Antheraea assama and Bombyx mori SFs. The morphology of the fibroin materials before and after coating with recombinant silk proteins was analyzed by scanning electron microscopy and atomic force microscopy. SF materials coated with various bioactive 4RepCT fusion proteins resulted in directed antibody capture, enzymatic activity, and improved cell attachment and spreading, respectively, compared to pristine SF materials. The herein-described procedure allows a fast and easy route for the construction of bioactive materials.

A poly(glycerol sebacate)-coated mesoporous bioactive glass scaffold with adjustable mechanical strength, degradation rate, controlled-release and cell behavior for bone tissue engineering.

Science.gov (United States)

Lin, Dan; Yang, Kai; Tang, Wei; Liu, Yutong; Yuan, Yuan; Liu, Changsheng

2015-07-01

Various requirements in the field of tissue engineering have motivated the development of three-dimensional scaffold with adjustable physicochemical properties and biological functions. A series of multiparameter-adjustable mesoporous bioactive glass (MBG) scaffolds with uncrosslinked poly(glycerol sebacate) (PGS) coating was prepared in this article. MBG scaffold was prepared by a modified F127/PU co-templating process and then PGS was coated by a simple adsorption and lyophilization process. Through controlling macropore parameters and PGS coating amount, the mechanical strength, degradation rate, controlled-release and cell behavior of the composite scaffold could be modulated in a wide range. PGS coating successfully endowed MBG scaffold with improved toughness and adjustable mechanical strength covering the bearing range of trabecular bone (2-12MPa). Multilevel degradation rate of the scaffold and controlled-release rate of protein from mesopore could be achieved, with little impact on the protein activity owing to an "ultralow-solvent" coating and "nano-cavity entrapment" immobilization method. In vitro studies indicated that PGS coating promoted cell attachment and proliferation in a dose-dependent manner, without affecting the osteogenic induction capacity of MBG substrate. These results first provide strong evidence that uncrosslinked PGS might also yield extraordinary achievements in traditional MBG scaffold. With the multiparameter adjustability, the composite MBG/PGS scaffolds would have a hopeful prospect in bone tissue engineering. The design considerations and coating method of this study can also be extended to other ceramic-based artificial scaffolds and are expected to provide new thoughts on development of future tissue engineering materials. Copyright © 2015 Elsevier B.V. All rights reserved.

Upregulated expression of integrin α1 in mesangial cells and integrin α3 and vimentin in podocytes of Col4a3-null (Alport mice.

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Brooke M Steenhard

Full Text Available Alport disease in humans, which usually results in proteinuria and kidney failure, is caused by mutations to the COL4A3, COL4A4, or COL4A5 genes, and absence of collagen α3α4α5(IV networks found in mature kidney glomerular basement membrane (GBM. The Alport mouse harbors a deletion of the Col4a3 gene, which also results in the lack of GBM collagen α3α4α5(IV. This animal model shares many features with human Alport patients, including the retention of collagen α1α2α1(IV in GBMs, effacement of podocyte foot processes, gradual loss of glomerular barrier properties, and progression to renal failure. To learn more about the pathogenesis of Alport disease, we undertook a discovery proteomics approach to identify proteins that were differentially expressed in glomeruli purified from Alport and wild-type mouse kidneys. Pairs of cy3- and cy5-labeled extracts from 5-week old Alport and wild-type glomeruli, respectively, underwent 2-dimensional difference gel electrophoresis. Differentially expressed proteins were digested with trypsin and prepared for mass spectrometry, peptide ion mapping/fingerprinting, and protein identification through database searching. The intermediate filament protein, vimentin, was upregulated ∼2.5 fold in Alport glomeruli compared to wild-type. Upregulation was confirmed by quantitative real time RT-PCR of isolated Alport glomeruli (5.4 fold over wild-type, and quantitative confocal immunofluorescence microscopy localized over-expressed vimentin specifically to Alport podocytes. We next hypothesized that increases in vimentin abundance might affect the basement membrane protein receptors, integrins, and screened Alport and wild-type glomeruli for expression of integrins likely to be the main receptors for GBM type IV collagen and laminin. Quantitative immunofluorescence showed an increase in integrin α1 expression in Alport mesangial cells and an increase in integrin α3 in Alport podocytes. We conclude that

Targeting of alpha-v integrins reduces malignancy of bladder carcinoma.

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Geertje van der Horst

Full Text Available Low survival rates of metastatic cancers emphasize the need for a drug that can prevent and/or treat metastatic cancer. αv integrins are involved in essential processes for tumor growth and metastasis and targeting of αv integrins has been shown to decrease angiogenesis, tumor growth and metastasis. In this study, the role of αv integrin and its potential as a drug target in bladder cancer was investigated. Treatment with an αv integrin antagonist as well as knockdown of αv integrin in the bladder carcinoma cell lines, resulted in reduced malignancy in vitro, as illustrated by decreased proliferative, migratory and clonogenic capacity. The CDH1/CDH2 ratio increased, indicating a shift towards a more epithelial phenotype. This shift appeared to be associated with downregulation of EMT-inducing transcription factors including SNAI2. The expression levels of the self-renewal genes NANOG and BMI1 decreased as well as the number of cells with high Aldehyde Dehydrogenase activity. In addition, self-renewal ability decreased as measured with the urosphere assay. In line with these observations, knockdown or treatment of αv integrins resulted in decreased metastatic growth in preclinical in vivo models as assessed by bioluminescence imaging. In conclusion, we show that αv integrins are involved in migration, EMT and maintenance of Aldehyde Dehydrogenase activity in bladder cancer cells. Targeting of αv integrins might be a promising approach for treatment and/or prevention of metastatic bladder cancer.

Alterated integrin expression in lichen planopilaris

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Erriquez Roberta

2007-02-01

Full Text Available Abstract Background Lichen planopilaris (LPP is an inflammatory disease characterized by a lymphomononuclear infiltrate surrounding the isthmus and infundibulum of the hair follicle of the scalp, that evolves into atrophic/scarring alopecia. In the active phase of the disease hairs are easily plucked with anagen-like hair-roots. In this study we focused on the expression of integrins and basement membrane components of the hair follicle in active LPP lesions. Methods Scalp biopsies were taken in 10 patients with LPP and in 5 normal controls. Using monoclonal antibodies against α3β1 and α6β4 integrins we showed the expression of these integrins and of the basement membrane components of the hair follicle in active LPP lesions and in healthy scalp skin. Results In the LPP involved areas, α3β1 was distributed in a pericellular pattern, the α6 subunit was present with a basolateral distribution while the β4 subunit showed discontinuous expression at the basal pole and occasionally, basolateral staining of the hair follicle. Conclusion: An altered distribution of the integrins in active LPP lesions can explain the phenomenon of easy pulling-out of the hair with a "gelatinous" root-sheath.

Characterization of new bioactive coatings of hydroxyapatite and TiO{sub 2} obtained by High-Velocity Oxy-Fuel; Caracterizacion de nuevos recubrimientos biocompatibles de hidroxiapatita-TiO{sub 2} obtenidos mediante Proyeccion Termica de Alta Velocidad

Energy Technology Data Exchange (ETDEWEB)

Melero, H.; Fernandez, J.; Dosta, S.; Guilemany, J. M.

2011-07-01

Hydroxyapatite (Hap: Ca{sub 1}0(PO{sub 4}){sub 6}OH{sub 2}) is a biocompatible and bioactive ceramic material widely used as a coating on metal surfaces (dental implants, hip replacements ...), but the low adhesion between Hap and the substrate, due to differences in thermal expansion coefficients of both (very important in thermal spraying because of the fast cooling of the coating, which can produce a lost of adherence), and the degradation of Hap, have been tried to be improved through the incorporation of TiO{sub 2} to get a good combination of mechanical properties. Therefore, the objective of this project is to produce coatings of Hap 80% TiO{sub 2} and 20% (by weight) on Ti6Al4V by High-Speed Thermal Spray (HVOF). The study of the microstructure has been carried out using scanning electron microscopy and characterization of the crystalline phases by X-ray diffraction and Raman spectrometry. The coatings adhesion has been measured by tensile tests according to ASTM C633-01 (2008), and their bioactivity also has been evaluated through its immersion in simulated body fluid (SBF), in order to measure their capacity to form an apatite layer on their surface. (Author) 26 refs.

Investigation of integrin expression on the surface of osteoblast-like cells by atomic force microscopy

International Nuclear Information System (INIS)

Caneva Soumetz, Federico; Saenz, Jose F.; Pastorino, Laura; Ruggiero, Carmelina; Nosi, Daniele; Raiteri, Roberto

2010-01-01

The transforming growth factor β1 (TGF-β1) is a human cytokine which has been demonstrated to modulate cell surface integrin repertoire. In this work integrin expression in response to TGF-β1 stimulation has been investigated on the surface of human osteoblast-like cells. We used atomic force microscopy (AFM) and confocal laser scanning microscopy to assess integrin expression and to evaluate their distribution over the dorsal side of the plasma membrane. AFM probes have been covalently functionalised with monoclonal antibodies specific to the β1 integrin subunit. Force curves have been collected in order to obtain maps of the interaction between the immobilized antibody and the respective cell membrane receptors. Adhesion peaks have been automatically detected by means of an ad hoc developed data analysis software. The specificity of the detected interactions has been assessed by adding free antibody in the solution and monitoring the dramatic decrease in the recorded interactions. In addition, the effect of TGF-β1 treatment on both the fluorescence signal and the adhesion events has been tested. The level of expression of the β1 integrin subunit was enhanced by TGF-β1. As a further analysis, the adhesion force of the single living cells to the substrate was measured by laterally pushing the cell with the AFM tip and measuring the force necessary to displace it. The treatment with TGF-β1 resulted in a decrease of the cell/substrate adhesion force. Results obtained by AFM have been validated by confocal laser scanning microscopy thus demonstrating the high potential of the AFM technique for the investigation of cell surface receptors distribution and trafficking at the nanoscale.

Investigation of integrin expression on the surface of osteoblast-like cells by atomic force microscopy

Energy Technology Data Exchange (ETDEWEB)

Caneva Soumetz, Federico [Department of Communication, Computer and System Sciences, University of Genova, Via Opera Pia, 13-16145 Genova (Italy); Saenz, Jose F. [Biophysical and Electronic Engineering Department, University of Genova, Via All' Opera Pia 11a, 16145 Genova (Italy); Pastorino, Laura; Ruggiero, Carmelina [Department of Communication, Computer and System Sciences, University of Genova, Via Opera Pia, 13-16145 Genova (Italy); Nosi, Daniele [Department of Anatomy, Histology and Forensic Medicine, Bio-photonic Laboratory, University of Florence, viale Morgagni, 85 Firenze, CAP 50134 Florence (Italy); Raiteri, Roberto, E-mail: rr@unige.it [Biophysical and Electronic Engineering Department, University of Genova, Via All' Opera Pia 11a, 16145 Genova (Italy)

2010-03-15

The transforming growth factor {beta}1 (TGF-{beta}1) is a human cytokine which has been demonstrated to modulate cell surface integrin repertoire. In this work integrin expression in response to TGF-{beta}1 stimulation has been investigated on the surface of human osteoblast-like cells. We used atomic force microscopy (AFM) and confocal laser scanning microscopy to assess integrin expression and to evaluate their distribution over the dorsal side of the plasma membrane. AFM probes have been covalently functionalised with monoclonal antibodies specific to the {beta}1 integrin subunit. Force curves have been collected in order to obtain maps of the interaction between the immobilized antibody and the respective cell membrane receptors. Adhesion peaks have been automatically detected by means of an ad hoc developed data analysis software. The specificity of the detected interactions has been assessed by adding free antibody in the solution and monitoring the dramatic decrease in the recorded interactions. In addition, the effect of TGF-{beta}1 treatment on both the fluorescence signal and the adhesion events has been tested. The level of expression of the {beta}1 integrin subunit was enhanced by TGF-{beta}1. As a further analysis, the adhesion force of the single living cells to the substrate was measured by laterally pushing the cell with the AFM tip and measuring the force necessary to displace it. The treatment with TGF-{beta}1 resulted in a decrease of the cell/substrate adhesion force. Results obtained by AFM have been validated by confocal laser scanning microscopy thus demonstrating the high potential of the AFM technique for the investigation of cell surface receptors distribution and trafficking at the nanoscale.

Biomimetic novel nanoporous niobium oxide coating for orthopaedic applications

Science.gov (United States)

Pauline, S. Anne; Rajendran, N.

2014-01-01

Niobium oxide was synthesized by sol-gel methodology and a crystalline, nanoporous and adherent coating of Nb2O5 was deposited on 316L SS using the spin coating technique and heat treatment. The synthesis conditions were optimized to obtain a nanoporous morphology. The coating was characterized using attenuated total reflectance-Infrared spectroscopy (ATR-IR), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX), atomic force microscopy (AFM) and transmission electron microscopy (TEM) and the formation of crystalline Nb2O5 coating with nanoporous morphology was confirmed. Mechanical studies confirmed that the coating has excellent adherence to the substrate and the hardness value of the coating was excellent. Contact angle analysis showed increased hydrophilicity for the coated substrate. In vitro bioactivity test confirmed that the Nb2O5 coating with nanoporous morphology facilitated the growth of hydroxyapatite (HAp). This was further confirmed by the solution analysis test where increased uptake of calcium and phosphorous ions from simulated body fluid (SBF) was observed. Electrochemical evaluation of the coating confirmed that the crystalline coating is insulative and protective in nature and offered excellent corrosion protection to 316L SS. Thus, this study confirmed that the nanoporous crystalline Nb2O5 coating conferred bioactivity and enhanced corrosion resistance on 316L SS.

Biomimetic novel nanoporous niobium oxide coating for orthopaedic applications

Energy Technology Data Exchange (ETDEWEB)

Pauline, S. Anne; Rajendran, N., E-mail: nrajendran@annauniv.edu

2014-01-30

Niobium oxide was synthesized by sol–gel methodology and a crystalline, nanoporous and adherent coating of Nb{sub 2}O{sub 5} was deposited on 316L SS using the spin coating technique and heat treatment. The synthesis conditions were optimized to obtain a nanoporous morphology. The coating was characterized using attenuated total reflectance-Infrared spectroscopy (ATR-IR), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX), atomic force microscopy (AFM) and transmission electron microscopy (TEM) and the formation of crystalline Nb{sub 2}O{sub 5} coating with nanoporous morphology was confirmed. Mechanical studies confirmed that the coating has excellent adherence to the substrate and the hardness value of the coating was excellent. Contact angle analysis showed increased hydrophilicity for the coated substrate. In vitro bioactivity test confirmed that the Nb{sub 2}O{sub 5} coating with nanoporous morphology facilitated the growth of hydroxyapatite (HAp). This was further confirmed by the solution analysis test where increased uptake of calcium and phosphorous ions from simulated body fluid (SBF) was observed. Electrochemical evaluation of the coating confirmed that the crystalline coating is insulative and protective in nature and offered excellent corrosion protection to 316L SS. Thus, this study confirmed that the nanoporous crystalline Nb{sub 2}O{sub 5} coating conferred bioactivity and enhanced corrosion resistance on 316L SS.

Evaluation of Moisture-Cure Urethane Coatings for Compliance with Industry Specifications

Science.gov (United States)

2011-12-01

polyurethane coating with a thermoset binder and micaceous iron oxide pigment reinforcement. Since SSPC paint specifications are designed for zinc ...Outside Coating System No.2 (minimum AWWA OFT 7.5 mils) 1 ct. Corothane I GalvaPac Zinc Primer@ 3.0 mils dft 1 ct. Corothane Iron ox B@ 3.0 mils dft...Substrates Over properly prepared: Ferrous Metal Galvanized Metal Aluminium/Non-Ferrous Metal Ductile Iron Previously Existing Coatings

Integrins as Therapeutic Targets: Successes and Cancers

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Sabine Raab-Westphal

2017-08-01

Full Text Available Integrins are transmembrane receptors that are central to the biology of many human pathologies. Classically mediating cell-extracellular matrix and cell-cell interaction, and with an emerging role as local activators of TGFβ, they influence cancer, fibrosis, thrombosis and inflammation. Their ligand binding and some regulatory sites are extracellular and sensitive to pharmacological intervention, as proven by the clinical success of seven drugs targeting them. The six drugs on the market in 2016 generated revenues of some US$3.5 billion, mainly from inhibitors of α4-series integrins. In this review we examine the current developments in integrin therapeutics, especially in cancer, and comment on the health economic implications of these developments.

Mechanisms of integrin-vascular endothelial growth factor receptor cross-activation in angiogenesis.

Science.gov (United States)

Mahabeleshwar, Ganapati H; Feng, Weiyi; Reddy, Kumar; Plow, Edward F; Byzova, Tatiana V

2007-09-14

The functional responses of endothelial cells are dependent on signaling from peptide growth factors and the cellular adhesion receptors, integrins. These include cell adhesion, migration, and proliferation, which, in turn, are essential for more complex processes such as formation of the endothelial tube network during angiogenesis. This study identifies the molecular requirements for the cross-activation between beta3 integrin and tyrosine kinase receptor 2 for vascular endothelial growth factor (VEGF) receptor (VEGFR-2) on endothelium. The relationship between VEGFR-2 and beta3 integrin appears to be synergistic, because VEGFR-2 activation induces beta3 integrin tyrosine phosphorylation, which, in turn, is crucial for VEGF-induced tyrosine phosphorylation of VEGFR-2. We demonstrate here that adhesion- and growth factor-induced beta3 integrin tyrosine phosphorylation are directly mediated by c-Src. VEGF-stimulated recruitment and activation of c-Src and subsequent beta3 integrin tyrosine phosphorylation are critical for interaction between VEGFR-2 and beta3 integrin. Moreover, c-Src mediates growth factor-induced beta3 integrin activation, ligand binding, beta3 integrin-dependent cell adhesion, directional migration of endothelial cells, and initiation of angiogenic programming in endothelial cells. Thus, the present study determines the molecular mechanisms and consequences of the synergism between 2 cell surface receptor systems, growth factor receptor and integrins, and opens new avenues for the development of pro- and antiangiogenic strategies.

Preparation and corrosion resistance of magnesium phytic acid/hydroxyapatite composite coatings on biodegradable AZ31 magnesium alloy.

Science.gov (United States)

Zhang, Min; Cai, Shu; Zhang, Feiyang; Xu, Guohua; Wang, Fengwu; Yu, Nian; Wu, Xiaodong

2017-06-01

In this work, a magnesium phytic acid/hydroxyapatite composite coating was successfully prepared on AZ31 magnesium alloy substrate by chemical conversion deposition technology with the aim of improving its corrosion resistance and bioactivity. The influence of hydroxyapatite (HA) content on the microstructure and corrosion resistance of the coatings was investigated. The results showed that with the increase of HA content in phytic acid solution, the cracks on the surface of the coatings gradually reduced, which subsequently improved the corrosion resistance of these coated magnesium alloy. Electrochemical measurements in simulated body fluid (SBF) revealed that the composite coating with 45 wt.% HA addition exhibited superior surface integrity and significantly improved corrosion resistance compared with the single phytic acid conversion coating. The results of the immersion test in SBF showed that the composite coating could provide more effective protection for magnesium alloy substrate than that of the single phytic acid coating and showed good bioactivity. Magnesium phytic acid/hydroxyapatite composite, with the desired bioactivity, can be synthesized through chemical conversion deposition technology as protective coatings for surface modification of the biodegradable magnesium alloy implants. The design idea of the new type of biomaterial is belong to the concept of "third generation biomaterial". Corrosion behavior and bioactivity of coated magnesium alloy are the key issues during implantation. In this study, preparation and corrosion behavior of magnesium phytic acid/hydroxyapatite composite coatings on magnesium alloy were studied. The basic findings and significance of this paper are as follows: 1. A novel environmentally friendly, homogenous and crack-free magnesium phytic acid/hydroxyapatite composite coating was fabricated on AZ31 magnesium alloy via chemical conversion deposition technology with the aim of enhancing its corrosion resistance and

Reduced graphene oxide-coated hydroxyapatite composites stimulate spontaneous osteogenic differentiation of human mesenchymal stem cells

Science.gov (United States)

Lee, Jong Ho; Shin, Yong Cheol; Jin, Oh Seong; Kang, Seok Hee; Hwang, Yu-Shik; Park, Jong-Chul; Hong, Suck Won; Han, Dong-Wook

2015-07-01

Human mesenchymal stem cells (hMSCs) have great potential as cell sources for bone tissue engineering and regeneration, but the control and induction of their specific differentiation into bone cells remain challenging. Graphene-based nanomaterials are considered attractive candidates for biomedical applications such as scaffolds in tissue engineering, substrates for SC differentiation and components of implantable devices, due to their biocompatible and bioactive properties. Despite the potential biomedical applications of graphene and its derivatives, only limited information is available regarding their osteogenic activity. This study concentrates upon the effects of reduced graphene oxide (rGO)-coated hydroxyapatite (HAp) composites on osteogenic differentiation of hMSCs. The average particle sizes of HAp and rGO were 1270 +/- 476 nm and 438 +/- 180 nm, respectively. When coated on HAp particulates, rGO synergistically enhanced spontaneous osteogenic differentiation of hMSCs, without hampering their proliferation. This result was confirmed by determining alkaline phosphatase activity and mineralization of calcium and phosphate as early and late stage markers of osteogenic differentiation. It is suggested that rGO-coated HAp composites can be effectively utilized as dental and orthopedic bone fillers since these graphene-based particulate materials have potent effects on stimulating the spontaneous differentiation of MSCs and show superior bioactivity and osteoinductive potential.Human mesenchymal stem cells (hMSCs) have great potential as cell sources for bone tissue engineering and regeneration, but the control and induction of their specific differentiation into bone cells remain challenging. Graphene-based nanomaterials are considered attractive candidates for biomedical applications such as scaffolds in tissue engineering, substrates for SC differentiation and components of implantable devices, due to their biocompatible and bioactive properties. Despite

Thermal analysis and in vitro bioactivity of bioactive glass-alumina composites

Energy Technology Data Exchange (ETDEWEB)

Chatzistavrou, Xanthippi, E-mail: x.chatzistavrou@imperial.ac.uk [Solid State Physics Section, Physics Department, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Kantiranis, Nikolaos, E-mail: kantira@geo.auth.gr [School of Geology, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Kontonasaki, Eleana, E-mail: kont@dent.auth.gr [School of Dentistry, Department of Fixed Prosthesis and Implant Prosthodontics, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Chrissafis, Konstantinos, E-mail: hrisafis@physics.auth.gr [Solid State Physics Section, Physics Department, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Papadopoulou, Labrini, E-mail: lambrini@geo.auth.gr [School of Geology, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Koidis, Petros, E-mail: pkoidis@dent.auth.gr [School of Dentistry, Department of Fixed Prosthesis and Implant Prosthodontics, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Boccaccini, Aldo R., E-mail: a.boccaccini@imperial.ac.uk [Department of Materials, Faculty of Engineering, Imperial College, SW7 2AZ London (United Kingdom); Paraskevopoulos, Konstantinos M., E-mail: kpar@auth.gr [Solid State Physics Section, Physics Department, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece)

2011-01-15

Bioactive glass-alumina composite (BA) pellets were fabricated in the range 95/5-60/40 wt.% respectively and were heat-treated under a specific thermal treatment up to 950 {sup o}C. Control (unheated) and heat-treated pellets were immersed in Simulated Body Fluid (SBF) for bioactivity testing. All pellets before and after immersion in SBF were studied by Fourier Transform Infrared spectroscopy (FTIR), Scanning Electron Microscopy (SEM-EDS) and X-ray Diffraction (XRD) analysis. All composite pellets presented bioactive response. On the surface of the heat-treated pellets the development of a rich biological hydroxyapatite (HAp) layer was delayed for one day, compared to the respective control pellets. Independent of the proportion of the two components, all composites of each group (control and heat-treated) presented the same bioactive response as a function of immersion time in SBF. It was found that by the applied methodology, Al{sub 2}O{sub 3} can be successfully applied in bioactive glass composites without obstructing their bioactive response. - Research Highlights: {yields} Isostatically pressed glass-alumina composites presented apatite-forming ability. {yields} The interaction with SBF resulted in an aluminium phosphate phase formation. {yields} The formation of an aluminium phosphate phase enhanced the in vitro apatite growth.

Construction and Biological Evaluation of a Novel Integrin ανβ3-Specific Carrier for Targeted siRNA Delivery In Vitro

Directory of Open Access Journals (Sweden)

Xueqi Chen

2017-02-01

Full Text Available (1 Background: The great potential of RNA interference (RNAi-based gene therapy is premised on the effective delivery of small interfering RNAs (siRNAs to target tissues and cells. Hence, we aimed at developing and examining a novel integrin αvβ3-specific delivery carrier for targeted transfection of siRNA to malignant tumor cells; (2 Methods: Arginine-glycine-aspartate motif (RGD was adopted as a tissue target for specific recognition of integrin αvβ3. To enable siRNA binding, a chimeric peptide was synthesized by adding nonamer arginine residues (9R at the carboxy terminus of cyclic-RGD dimer, designated as c(RGD2-9R. The efficiency of 9R peptide transferring siRNA was biologically evaluated in vitro by flow cytometry, confocal microscopy, and Western blot; (3 Results: An optimal 10:1 molar ratio of c(RGD2-9R to siRNA was confirmed by the electrophoresis on agarose gels. Both the flow cytometry and confocal microscopy results testified that transfection of c(RGD2-9R as an siRNA delivery carrier was obviously higher than the naked-siRNA group. The results of Western blot demonstrated that these 9R peptides were able to transduce siRNA to HepG2 cells in vitro, resulting in efficient gene silencing; and (4 Conclusion: The chimeric peptide of c(RGD2-9R can be developed as an effective siRNA delivery carrier and shows potential as a new strategy for RNAi-based gene therapy.

Exosomes derived from human macrophages suppress endothelial cell migration by controlling integrin trafficking.

Science.gov (United States)

Lee, Hee Doo; Kim, Yeon Hyang; Kim, Doo-Sik

2014-04-01

Integrin trafficking, including internalization, recycling, and lysosomal degradation, is crucial for the regulation of cellular functions. Exosomes, nano-sized extracellular vesicles, are believed to play important roles in intercellular communications. This study demonstrates that exosomes released from human macrophages negatively regulate endothelial cell migration through control of integrin trafficking. Macrophage-derived exosomes promote internalization of integrin β1 in primary HUVECs. The internalized integrin β1 persistently accumulates in the perinuclear region and is not recycled back to the plasma membrane. Experimental results indicate that macrophage-derived exosomes stimulate trafficking of internalized integrin β1 to lysosomal compartments with a corresponding decrease in the integrin destined for recycling endosomes, resulting in proteolytic degradation of the integrin. Moreover, ubiquitination of HUVEC integrin β1 is enhanced by the exosomes, and exosome-mediated integrin degradation is blocked by bafilomycin A, a lysosomal degradation inhibitor. Macrophage-derived exosomes were also shown to effectively suppress collagen-induced activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase signaling pathway and HUVEC migration, which are both dependent on integrin β1. These observations provide new insight into the functional significance of exosomes in the regulation of integrin trafficking. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Alpha1 and Alpha2 Integrins Mediate Invasive Activity of Mouse Mammary Carcinoma Cells through Regulation of Stromelysin-1 Expression

Energy Technology Data Exchange (ETDEWEB)

Lochter, Andre; Navre, Marc; Werb, Zena; Bissell, Mina J

1998-06-29

Tumor cell invasion relies on cell migration and extracellular matrix proteolysis. We investigated the contribution of different integrins to the invasive activity of mouse mammary carcinoma cells. Antibodies against integrin subunits {alpha}6 and {beta}1, but not against {alpha}1 and {alpha}2, inhibited cell locomotion on a reconstituted basement membrane in two-dimensional cell migration assays, whereas antibodies against {beta}1, but not against a6 or {alpha}2, interfered with cell adhesion to basement membrane constituents. Blocking antibodies against {alpha}1 integrins impaired only cell adhesion to type IV collagen. Antibodies against {alpha}1, {alpha}2, {alpha}6, and {beta}1, but not {alpha}5, integrin subunits reduced invasion of a reconstituted basement membrane. Integrins {alpha}1 and {alpha}2, which contributed only marginally to motility and adhesion, regulated proteinase production. Antibodies against {alpha}1 and {alpha}2, but not {alpha}6 and {beta}1, integrin subunits inhibited both transcription and protein expression of the matrix metalloproteinase stromelysin-1. Inhibition of tumor cell invasion by antibodies against {alpha}1 and {alpha}2 was reversed by addition of recombinant stromelysin-1. In contrast, stromelysin-1 could not rescue invasion inhibited by anti-{alpha}6 antibodies. Our data indicate that {alpha}1 and {alpha}2 integrins confer invasive behavior by regulating stromelysin-1 expression, whereas {alpha}6 integrins regulate cell motility. These results provide new insights into the specific functions of integrins during tumor cell invasion.

Functional consequences of integrin gene mutations in mice

DEFF Research Database (Denmark)

Bouvard, D; Brakebusch, C; Gustafsson, E

2001-01-01

Integrins are cell-surface receptors responsible for cell attachment to extracellular matrices and to other cells. The application of mouse genetics has significantly increased our understanding of integrin function in vivo. In this review, we summarize the phenotypes of mice carrying mutant inte...

Microstructure and osteoblast response of gradient bioceramic coating on titanium alloy fabricated by laser cladding

International Nuclear Information System (INIS)

Zheng Min; Fan Ding; Li Xiukun; Li Wenfei; Liu Qibin; Zhang Jianbin

2008-01-01

To construct a bioactive interface between metal implant and the surrounding bone tissue, the gradient calcium phosphate bioceramic coating on titanium alloy (Ti-6Al-4V) was designed and fabricated by laser cladding. The results demonstrated that the gradient bioceramic coating was metallurgically bonded to the titanium alloy substrate. The appearance of hydroxyapatite and β-tricalcium phosphate indicated that the bioactive phases were synthesized on the surface of coating. The microhardness gradually decreased from the coating to substrate, which could help stress relaxation between coating and bone tissue. Furthermore, the methyl thiazolyl tetrazolium (MTT) assay of cell proliferation revealed that the laser-cladded bioceramic coating had more favorable osteoblast response compared with the surface of untreated titanium alloy substrate

Microstructure and osteoblast response of gradient bioceramic coating on titanium alloy fabricated by laser cladding

Energy Technology Data Exchange (ETDEWEB)

Zheng Min [State Key Laboratory of Gansu Advanced Non-ferrous Metal Materials, Lanzhou University of Technology, Lanzhou 730050 (China)], E-mail: zhminmin@sina.com; Fan Ding; Li Xiukun [State Key Laboratory of Gansu Advanced Non-ferrous Metal Materials, Lanzhou University of Technology, Lanzhou 730050 (China); Li Wenfei; Liu Qibin [College of Materials Science and Engineering, Guizhou University, Guiyang 550003 (China); Zhang Jianbin [State Key Laboratory of Gansu Advanced Non-ferrous Metal Materials, Lanzhou University of Technology, Lanzhou 730050 (China)

2008-11-15

To construct a bioactive interface between metal implant and the surrounding bone tissue, the gradient calcium phosphate bioceramic coating on titanium alloy (Ti-6Al-4V) was designed and fabricated by laser cladding. The results demonstrated that the gradient bioceramic coating was metallurgically bonded to the titanium alloy substrate. The appearance of hydroxyapatite and {beta}-tricalcium phosphate indicated that the bioactive phases were synthesized on the surface of coating. The microhardness gradually decreased from the coating to substrate, which could help stress relaxation between coating and bone tissue. Furthermore, the methyl thiazolyl tetrazolium (MTT) assay of cell proliferation revealed that the laser-cladded bioceramic coating had more favorable osteoblast response compared with the surface of untreated titanium alloy substrate.

Strontium and magnesium substituted dicalcium phosphate dehydrate coating for carbon/carbon composites prepared by pulsed electrodeposition

Energy Technology Data Exchange (ETDEWEB)

Liu, Shou-jie, E-mail: jlliushoujie@126.com; Li, He-jun, E-mail: lihejun@nwpu.edu.cn; Zhang, Lei-lei, E-mail: zhangleilei@nwpu.edu.cn; Feng, Lei, E-mail: fengleijinan@163.com; Yao, Pei, E-mail: 1113923884@qq.com

2015-12-30

Graphical abstract: The potentiodynamic polarization curve shows that the SM-DCPD coating can dramatically enhance the corrosion potential (E{sub corr}) value and meanwhile decrease the corrosion current density (I{sub corr}) of C/C composites. - Highlights: • Strontium and magnesium substituted dicalcium phosphate dehydrate coatings for carbon/carbon composites were synthesized by pulsed eletrodeposition. • Strontium and magnesium substituted dicalcium phosphate dehydrate coated carbon/carbon composites exhibited excellent bioactivity in vivo. • Strontium and magnesium substituted dicalcium phosphate dehydrate coated carbon/carbon composites showed lower corrosion rate with the comparison to pure carbon/carbon composites. - Abstract: Trace elements substituted apatite coatings have received a lot of interest recently as they have many benefits. In this work, strontium and magnesium substituted DCPD (SM-DCPD) coatings were deposited on carbon/carbon (C/C) composites by pulsed electrodeposition method. The morphology, microstructure, corrosion resistance and in vitro bioactivity of the SM-DCPD coatings are analyzed. The results show that the SM-DCPD coatings exhibit a flake-like morphology with dense and uniform structure. The SM-DCPD coatings could induce the formation of apatite layers on their surface in simulated body fluid. The electrochemical test indicates that the SM-DCPD coatings can evidently decrease the corrosion rate of the C/C composites in simulated body fluid. The SM-DCPD has potential application as the bioactive coatings.

Synthesis and biological evaluation of potent αvβ3-integrin receptor antagonists

International Nuclear Information System (INIS)

Dijkgraaf, Ingrid; Kruijtzer, John A.W.; Frielink, Cathelijne; Soede, Annemieke C.; Hilbers, Hans W.; Oyen, Wim J.G.; Corstens, Frans H.M.; Liskamp, Rob M.J.; Boerman, Otto C.

2006-01-01

Introduction: α v β 3 Integrin is expressed in sprouting endothelial cells in growing tumors, whereas it is absent in quiescent blood vessels. In addition, various tumor cell types express α v β 3 integrin. α v β 3 Integrin, a transmembrane heterodimeric protein, binds to the arginine-glycine-aspartic acid (RGD) amino acid sequence of extracellular matrix proteins such as vitronectin and plays a pivotal role in invasion, proliferation and metastasis. Due to the selective expression of α v β 3 integrin in tumors, radiolabeled RGD peptides and peptidomimetics are attractive candidates for tumor targeting. Methods: A cyclic RGD peptide, a peptoid-peptide hybrid, an all-peptoid and a peptidomimetic compound were synthesized, conjugated with 1,4,7,10-tetraazadodecane-N,N',N'',N'''-tetraacetic acid (DOTA) and radiolabeled with 111 In. Their in vitro and in vivo α v β 3 -binding characteristics were determined. Results: IC 5 values were 236 nM for DOTA-E-c(RGDfK), 219 nM for DOTA-peptidomimetic, >10 mM for DOTA-all-peptoid and 9.25 mM for the peptoid-peptide hybrid DOTA-E-c(nRGDfK). 111 In-labeled compounds, except for [ 111 In]DOTA-all-peptoid, showed specific uptake in human α v β 3 -expressing tumors xenografted in athymic mice. Tumor uptake for [ 111 In]DOTA-E-c(RGDfK) was 1.73±0.4% ID/g (2 h postinjection) and that of [ 111 In]DOTA-peptidomimetic was 2.04±0.3% ID/g. Tumor uptake for the peptoid-peptide hybrid [ 111 In]DOTA-E-c(nRGDfK) was markedly lower (0.45±0.07% ID/g). The all-peptoid [ 111 In]DOTA-E-c(nRGnDnFnK) did not show specific uptake in tumors (0.11±0.04% ID/g). Conclusions: The peptidomimetic compound and the cyclic RGD peptide have a high affinity for α v β 3 integrin, and these compounds have better tumor-targeting characteristics than the peptoid-peptide hybrid and the all-peptoid

Endothelial cell-derived matrix promotes the metabolic functional maturation of hepatocyte via integrin-Src signalling.

Science.gov (United States)

Guo, Xinyue; Li, Weihong; Ma, Minghui; Lu, Xin; Zhang, Haiyan

2017-11-01

The extracellular matrix (ECM) microenvironment is involved in the regulation of hepatocyte phenotype and function. Recently, the cell-derived extracellular matrix has been proposed to represent the bioactive and biocompatible materials of the native ECM. Here, we show that the endothelial cell-derived matrix (EC matrix) promotes the metabolic maturation of human adipose stem cell-derived hepatocyte-like cells (hASC-HLCs) through the activation of the transcription factor forkhead box protein A2 (FOXA2) and the nuclear receptors hepatocyte nuclear factor 4 alpha (HNF4α) and pregnane X receptor (PXR). Reducing the fibronectin content in the EC matrix or silencing the expression of α5 integrin in the hASC-HLCs inhibited the effect of the EC matrix on Src phosphorylation and hepatocyte maturation. The inhibition of Src phosphorylation using the inhibitor PP2 or silencing the expression of Src in hASC-HLCs also attenuated the up-regulation of the metabolic function of hASC-HLCs in a nuclear receptor-dependent manner. These data elucidate integrin-Src signalling linking the extrinsic EC matrix signals and metabolic functional maturation of hepatocyte. This study provides a model for studying the interaction between hepatocytes and non-parenchymal cell-derived matrix. © 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Retinoic acid receptor gamma impacts cellular adhesion, Alpha5Beta1 integrin expression and proliferation in K562 cells.

Science.gov (United States)

Kelley, Melissa D; Phomakay, Raynin; Lee, Madison; Niedzwiedz, Victoria; Mayo, Rachel

2017-01-01

The interplay between cellular adhesion and proliferation is complex; however, integrins, particularly the α5β1 subset, play a pivotal role in orchestrating critical cellular signals that culminate in cellular adhesion and growth. Retinoids modify the expression of a variety of adhesive/proliferative signaling proteins including α5β1 integrins; however, the role of specific retinoic acid receptors involved in these processes has not been elucidated. In this study, the effect of all-trans-retinoic acid receptor (RAR) agonists on K562 cellular adhesion, proliferation, and α5β1 integrin cell surface expression was investigated. RARγ agonist exposure increased K562 cellular adhesion to RGD containing extracellular matrix proteins fibronectin and FN-120 in a time- and concentration dependent manner, while RARα or RARβ agonist treatment had no effect on cellular adhesion. Due to the novel RARγ- dependent cellular adhesion response exhibited by K562 cells, we examined α5 and β1 integrin subunit expression when K562 cells were exposed to retinoid agonists or vehicle for 24, 48, 72 or 96 hours. Our data demonstrates no differences in K562 cell surface expression of the α5 integrin subunit when cells were exposed to RARα, RARβ, or RARγ agonists for all time points tested. In contrast, RARγ agonist exposure resulted in an increase in cell surface β1 integrin subunit expression within 48 hours that was sustained at 72 and 96 hours. Finally, we demonstrate that while exposure to RARα or RARβ agonists have no effect on K562 cellular proliferation, the RARγ agonist significantly dampens K562 cellular proliferation levels in a time- and concentration- dependent manner. Our study is the first to report that treatment with a RARγ specific agonist augments cellular adhesion to α5β1 integrin substrates, increases cell surface levels of the β1 integrin subunit, and dampens cellular proliferation in a time and concentration dependent manner in a human

Hydroxyapatite coating on stainless steel by biomimetic method

International Nuclear Information System (INIS)

Dias, V.M.; Maia Filho, A.L.M.; Silva, G.; Sousa, E. de; Cardoso, K.R.

2010-01-01

Austenitic stainless steels are widely used in implants due to their high mechanical strength and corrosion, however, are not able to connect to bone tissue and were classified as bioinert. The calcium phosphate ceramics such as hydroxyapatite (HA) are bioactive materials and create strong chemical bonds with bone tissue, but its brittleness and low fracture toughness render its use in conditions of high mechanical stress. The coating of steel with the bioactive ceramics such as HA, combines the properties of interest of both materials, accelerating bone formation around the implant. In this study, austenitic stainless steel samples were coated with apatite using the biomimetic method. The effect of three different surface conditions of steel and the immersion time in the SBF solution on the coating was evaluated. The samples were characterized by SEM, EDS and X-ray diffraction. (author)

Gelatin Tight-Coated Poly(lactide-co-glycolide Scaffold Incorporating rhBMP-2 for Bone Tissue Engineering

Directory of Open Access Journals (Sweden)

Juan Wang

2015-03-01

Full Text Available Surface coating is the simplest surface modification. However, bioactive molecules can not spread well on the commonly used polylactone-type skeletons; thus, the surface coatings of biomolecules are typically unstable due to the weak interaction between the polymer and the bioactive molecules. In this study, a special type of poly(lactide-co-glycolide (PLGA-based scaffold with a loosened skeleton was fabricated by phase separation, which allowed gelatin molecules to more readily diffuse throughout the structure. In this application, gelatin modified both the internal substrate and external surface. After cross-linking with glutaraldehyde, the surface layer gelatin was tightly bound to the diffused gelatin, thereby preventing the surface layer gelatin coating from falling off within 14 days. After gelatin modification, PLGA scaffold demonstrated enhanced hydrophilicity and improved mechanical properties (i.e., increased compression strength and elastic modulus in dry and wet states. Furthermore, a sustained release profile of recombinant human bone morphogenetic protein-2 (rhBMP-2 was achieved in the coated scaffold. The coated scaffold also supported the in vitro attachment, proliferation, and osteogenesis of rabbit bone mesenchymal stem cells (BMSCs, indicating the bioactivity of rhBMP-2. These results collectively demonstrate that the cross-linked-gelatin-coated porous PLGA scaffold incorporating bioactive molecules is a promising candidate for bone tissue regeneration.

Catfish thrombocytes express an integrin-like CD41/CD61 complex.

Science.gov (United States)

Passer, B J; Chen, C H; Miller, N W; Cooper, M D

1997-08-01

A thrombocyte-specific antigen was identified in two closely related catfish, Ictalurus punctatus and Ictalurus furcatus, by monoclonal antibodies 4-20 and 7-2. The antibodies immunoprecipitate two noncovalently associated glycoprotein chains of Mr 180,000 and Mr 95,000. Under reducing conditions the Mr 180,000 chain is resolved into Mr 150,000 and 32,000 subcomponents. Analysis of N-terminal amino acid sequences indicates homology of the Mr 95,000 chain with the beta3 integrin subunit and homology of the Mr 150,000 chain with the alphaIIb integrin subunit. These antibodies induce catfish thrombocyte aggregation and alteration of cell shape. The data indicate conservation of the megakaryocyte/platelet-restricted CD41/CD61 complex in bony fish.

Integrin β1, osmosensing, and chemoresistance in mouse ehrlich carcinoma cells

DEFF Research Database (Denmark)

Sørensen, Belinda Halling; Rasmussen, Line Jee Hartmann; Broberg, Bjørn Sindballe

2015-01-01

BACKGROUND/AIMS: Altered expression of the integrin family of cell adhesion receptors has been associated with initiation, progression, and metastasis of solid tumors as well as in the development of chemoresistance. Here, we investigated the role of integrins, in particular integrin β1, in cell ...

Integrin αv in the mechanical response of osteoblast lineage cells

Energy Technology Data Exchange (ETDEWEB)

Kaneko, Keiko [Department of Bone and Joint Disease, National Center for Geriatrics and Gerontology, Obu, Aichi 474-8511 (Japan); Ito, Masako [Medical Work-Life-Balance Center, Nagasaki University Hospital, Nagasaki 852-8501 (Japan); Naoe, Yoshinori [Department of Mechanism of Aging, National Center for Geriatrics and Gerontology, Obu, Aichi 474-8511 (Japan); Lacy-Hulbert, Adam [Department of Pediatrics, Massachusetts General Hospital, Boston, MA 02114 (United States); Ikeda, Kyoji, E-mail: kikeda@ncgg.go.jp [Department of Bone and Joint Disease, National Center for Geriatrics and Gerontology, Obu, Aichi 474-8511 (Japan)

2014-05-02

Highlights: • Deletion of integrin αv in osteoblast lineage results in an impaired SOST response to loading in vivo. • c-Src–p130Cas–JNK–YAP/TAZ is activated via integrin αv on osteoblasts in response to FSS. • Deletion of integrin αv in osteoblasts results in impaired responses to mechanical stimulation. • Integrin αv is a key component of the mechanosensing machinery in bone. - Abstract: Although osteoblast lineage cells, especially osteocytes, are thought to be a primary mechanosensory cell in bone, the identity of the mechano-receptor and downstream mechano-signaling pathways remain largely unknown. Here we show using osteoblastic cell model of mechanical stimulation with fluid shear stress that in the absence of integrin αv, phosphorylation of the Src substrate p130Cas and JNK was impaired, culminating in an inhibition of nuclear translocation of YAP/TAZ and subsequent transcriptional activation of target genes. Targeted deletion of the integrin αv in osteoblast lineage cells results in an attenuated response to mechanical loading in terms of Sost gene expression, indicative of a role for integrin αv in mechanoreception in vivo. Thus, integrin αv may be integral to a mechanosensing machinery in osteoblastic cells and involved in activation of a Src–JNK–YAP/TAZ pathway in response to mechanical stimulation.

Integrin αv in the mechanical response of osteoblast lineage cells

International Nuclear Information System (INIS)

Kaneko, Keiko; Ito, Masako; Naoe, Yoshinori; Lacy-Hulbert, Adam; Ikeda, Kyoji

2014-01-01

Highlights: • Deletion of integrin αv in osteoblast lineage results in an impaired SOST response to loading in vivo. • c-Src–p130Cas–JNK–YAP/TAZ is activated via integrin αv on osteoblasts in response to FSS. • Deletion of integrin αv in osteoblasts results in impaired responses to mechanical stimulation. • Integrin αv is a key component of the mechanosensing machinery in bone. - Abstract: Although osteoblast lineage cells, especially osteocytes, are thought to be a primary mechanosensory cell in bone, the identity of the mechano-receptor and downstream mechano-signaling pathways remain largely unknown. Here we show using osteoblastic cell model of mechanical stimulation with fluid shear stress that in the absence of integrin αv, phosphorylation of the Src substrate p130Cas and JNK was impaired, culminating in an inhibition of nuclear translocation of YAP/TAZ and subsequent transcriptional activation of target genes. Targeted deletion of the integrin αv in osteoblast lineage cells results in an attenuated response to mechanical loading in terms of Sost gene expression, indicative of a role for integrin αv in mechanoreception in vivo. Thus, integrin αv may be integral to a mechanosensing machinery in osteoblastic cells and involved in activation of a Src–JNK–YAP/TAZ pathway in response to mechanical stimulation

Bioactivity and Osseointegration of PEEK Are Inferior to Those of Titanium: A Systematic Review.

Science.gov (United States)

Najeeb, Shariq; Bds, Zohaib Khurshid; Bds, Sana Zohaib; Bds, Muhammad Sohail Zafar

2016-12-01

Polyetheretherketone (PEEK) has been suggested as an alternative to replace titanium as a dental implant material. However, PEEK's bioactivity and osseointegration are debatable. This review has systematically analyzed studies that have compared PEEK (or PEEK-based) implants with titanium implants so that its feasibility as a possible replacement for titanium can be determined. The focused question was: "Are the bioactivity and osseointegration of PEEK implants comparable to or better than titanium implants?" Using the key words "dental implant," "implant," "polyetheretherketone," "PEEK," and "titanium" in various combinations, the following databases were searched electronically: PubMED/MEDLINE, Embase, Google Scholar, ISI Web of Knowledge, and Cochrane Database. 5 in vitro and 4 animal studies were included in the review. In 4 out of 5 in vitro studies, titanium exhibited more cellular proliferation, angiogenesis, osteoblast maturation, and osteogenesis compared to PEEK; one in vitro study observed comparable outcomes regardless of the implant material. In all animal studies, uncoated and coated titanium exhibited a more osteogenic behavior than did uncoated PEEK, while comparable bone-implant contact was observed in HA-coated PEEK and coated titanium implants. Unmodified PEEK is less osseoconductive and bioactive than titanium. Furthermore, the majority of studies had multiple sources of bias; hence, in its unmodified form, PEEK is unsuitable to be used as dental implant. Significantly more research and long-term trials must focus on improving the bioactivity of PEEK before it can be used as dental implant. More comparative animal and clinical studies are warranted to ascertain the potential of PEEK as a viable alternative to titanium.

Synthesis and biological evaluation of potent alphavbeta3-integrin receptor antagonists.

NARCIS (Netherlands)

Dijkgraaf, I.; Kruijtzer, J.A.; Frielink, C.; Soede, A.C.; Hilbers, H.W.; Oyen, W.J.G.; Corstens, F.H.M.; Liskamp, R.M.; Boerman, O.C.

2006-01-01

INTRODUCTION: alpha(v)beta(3) Integrin is expressed in sprouting endothelial cells in growing tumors, whereas it is absent in quiescent blood vessels. In addition, various tumor cell types express alpha(v)beta(3) integrin. alpha(v)beta(3) Integrin, a transmembrane heterodimeric protein, binds to the

LASER-INDUCED BIOACTIVITY IN DENTAL PORCELAIN MODIFIED BY BIOACTIVE GLASS

Directory of Open Access Journals (Sweden)

ANASTASIA BEKETOVA

2012-12-01

Full Text Available The aim of this study was to investigate the impact of laser-liquid-solid interaction method in the bioactivity of dental porcelain modified by bioactive glass. Forty sol-gel derived specimens were immersed in Dulbecco's Modified Eagle's Medium, 31 and 9 specimens of which were treated with Er:YAG and Nd:YAG laser respectively. Untreated specimens served as controls. Incubation of specimens followed. Bioactivity was evaluated, using Fourier Transform Infrared spectroscopy (FTIR, Scanning Electron Microscopy (SEM/Energy Dispersive Spectroscopy (EDS and Transmission Electron Microscopy (TEM. FTIR detected peaks associated with hydroxyapatite on 1 Nd:YAG- and 4 Er:YAG-treated specimens. SEM analysis revealed that Er:YAG-treated specimens were covered by granular hydroxyapatite layer, while Nd:YAG treated specimen presented growth of flake-like hydroxyapatite. TEM confirmed the results. The untreated controls presented delayed bioactivity. In conclusion, Nd:YAG and Er:YAG laser treatment of the material, under certain fluencies, accelerates hydroxyapatite formation. Nd:YAG laser treatment of specific parameters causes the precipitation of flake-like hydroxyapatite in nano-scale.

Tissue inhibitor of metalloproteinase-2 (TIMP-2) regulates myogenesis and β1 integrin expression in vitro

International Nuclear Information System (INIS)

Lluri, Gentian; Langlois, Garret D.; Soloway, Paul D.; Jaworski, Diane M.

2008-01-01

Myogenesis in vitro involves myoblast cell cycle arrest, migration, and fusion to form multinucleated myotubes. Extracellular matrix (ECM) integrity during these processes is maintained by the opposing actions of matrix metalloproteinase (MMP) proteases and their inhibitors, the tissue inhibitor of metalloproteinases (TIMPs). Here, we report that TIMP-2, MMP-2, and MT1-MMP are differentially expressed during mouse myoblast differentiation in vitro. A specific role for TIMP-2 in myogenesis is demonstrated by altered TIMP-2 -/- myotube formation. When differentiated in horse serum-containing medium, TIMP-2 -/- myotubes are larger than wild-type myotubes. In contrast, when serum-free medium is used, TIMP-2 -/- myotubes are smaller than wild-type myotubes. Regardless of culture condition, myotube size is directly correlated with MMP activity and inversely correlated with β1 integrin expression. Treatment with recombinant TIMP-2 rescues reduced TIMP-2 -/- myotube size and induces increased MMP-9 activation and decreased β1 integrin expression. Treatment with either MMP-2 or MMP-9 similarly rescues reduced myotube size, but has no effect on β1 integrin expression. These data suggest a specific regulatory relationship between TIMP-2 and β1 integrin during myogenesis. Elucidating the role of TIMP-2 in myogenesis in vitro may lead to new therapeutic options for the use of TIMP-2 in myopathies and muscular dystrophies in vivo

The Gain-of-Function Integrin β3 Pro33 Variant Alters the Serotonin System in the Mouse Brain.

Science.gov (United States)

Dohn, Michael R; Kooker, Christopher G; Bastarache, Lisa; Jessen, Tammy; Rinaldi, Capria; Varney, Seth; Mazalouskas, Matthew D; Pan, Hope; Oliver, Kendra H; Velez Edwards, Digna R; Sutcliffe, James S; Denny, Joshua C; Carneiro, Ana M D

2017-11-15

Engagement of integrins by the extracellular matrix initiates signaling cascades that drive a variety of cellular functions, including neuronal migration and axonal pathfinding in the brain. Multiple lines of evidence link the ITGB3 gene encoding the integrin β3 subunit with the serotonin (5-HT) system, likely via its modulation of the 5-HT transporter (SERT). The ITGB3 coding polymorphism Leu33Pro (rs5918, Pl A2 ) produces hyperactive αvβ3 receptors that influence whole-blood 5-HT levels and may influence the risk for autism spectrum disorder (ASD). Using a phenome-wide scan of psychiatric diagnoses, we found significant, male-specific associations between the Pro33 allele and attention-deficit hyperactivity disorder and ASDs. Here, we used knock-in (KI) mice expressing an Itgb3 variant that phenocopies the human Pro33 variant to elucidate the consequences of constitutively enhanced αvβ3 signaling to the 5-HT system in the brain. KI mice displayed deficits in multiple behaviors, including anxiety, repetitive, and social behaviors. Anatomical studies revealed a significant decrease in 5-HT synapses in the midbrain, accompanied by decreases in SERT activity and reduced localization of SERTs to integrin adhesion complexes in synapses of KI mice. Inhibition of focal adhesion kinase (FAK) rescued SERT function in synapses of KI mice, demonstrating that constitutive active FAK signaling downstream of the Pro32Pro33 integrin αvβ3 suppresses SERT activity. Our studies identify a complex regulation of 5-HT homeostasis and behaviors by integrin αvβ3, revealing an important role for integrins in modulating risk for neuropsychiatric disorders. SIGNIFICANCE STATEMENT The integrin β3 Leu33Pro coding polymorphism has been associated with autism spectrum disorders (ASDs) within a subgroup of patients with elevated blood 5-HT levels, linking integrin β3, 5-HT, and ASD risk. We capitalized on these interactions to demonstrate that the Pro33 coding variation in the murine

Influence of Nano-HA Coated Bone Collagen to Acrylic (Polymethylmethacrylate Bone Cement on Mechanical Properties and Bioactivity.

Directory of Open Access Journals (Sweden)

Tao Li

Full Text Available This research investigated the mechanical properties and bioactivity of polymethylmethacrylate (PMMA bone cement after addition of the nano-hydroxyapatite(HA coated bone collagen (mineralized collagen, MC.The MC in different proportions were added to the PMMA bone cement to detect the compressive strength, compression modulus, coagulation properties and biosafety. The MC-PMMA was embedded into rabbits and co-cultured with MG 63 cells to exam bone tissue compatibility and gene expression of osteogenesis.15.0%(wt impregnated MC-PMMA significantly lowered compressive modulus while little affected compressive strength and solidification. MC-PMMA bone cement was biologically safe and indicated excellent bone tissue compatibility. The bone-cement interface crosslinking was significantly higher in MC-PMMA than control after 6 months implantation in the femur of rabbits. The genes of osteogenesis exhibited significantly higher expression level in MC-PMMA.MC-PMMA presented perfect mechanical properties, good biosafety and excellent biocompatibility with bone tissues, which has profoundly clinical values.

Bitistatin-functionalized fluorescent nanodiamond particles specifically bind to purified human platelet integrin receptor αIIbβ3 and activated platelets.

Science.gov (United States)

Marcinkiewicz, Cezary; Gerstenhaber, Jonathan A; Sternberg, Mark; Lelkes, Peter I; Feuerstein, Giora

2017-01-01

Thromboembolic events (TEE) underwrite key causes of death in developed countries. While advanced imaging technologies such as computed tomography scans serve to diagnose blood clots during acute cardiovascular events, no such technology is available in routine primary care for TEE risk assessment. Here, we describe an imaging platform technology based on bioengineered fluorescent nanodiamond particles (F-NDPs) functionalized with bitistatin (Bit), a disintegrin that specifically binds to the α IIb β 3 integrin, platelet fibrinogen receptor (PFR) on activated platelets. Covalent linkage of purified Bit to F-NDP was concentration-dependent and saturable, as validated by enzyme-linked immunosorbent assay using specific anti-Bit antibodies. F-NDP-Bit interacted with purified PFR, either in immobilized or soluble form. Lotrafiban, a nonpeptide, α IIb β 3 receptor antagonist, specifically blocked F-NDP-Bit-PFR complex formation. Moreover, F-NDP-Bit specifically binds to activated platelets incorporated into a clot generated by thrombin-activated rat platelet-rich plasma (PRP). Our results suggest that engineered F-NDP-Bit particles could serve as noninvasive, "real-time" optical diagnostics for clots present in blood vessels.

Osteopontin binding to the alpha 4 integrin requires highest affinity integrin conformation, but is independent of post-translational modifications of osteopontin

DEFF Research Database (Denmark)

Hui, Tommy; Sørensen, Esben Skipper; Rittling, Susan R.

2015-01-01

Osteopontin (OPN) is a ligand for the α4 integrin, but the physiological importance of this binding is not well understood. Here, we have assessed the effect of posttranslational modifications on OPN binding to the α4 integrin on cultured human leukocyte cell lines, and compared OPN interaction...

Single-walled carbon nanotubes/hydroxyapatite coatings on titanium obtained by electrochemical deposition

International Nuclear Information System (INIS)

Pei, Xibo; Zeng, Yongxiang; He, Rui; Li, Zhongjie; Tian, Lingyang; Wang, Jian; Wan, Qianbing; Li, Xiaoyu; Bao, Hong

2014-01-01

Graphical abstract: - Highlights: • The incorporation of SWNTs into the HA coating leaded to the formation of homogeneous and crack-free composite coatings. • The highest bonding strength was detected for the SWNTs/HA-0.5 composite coating (25.70 MPa). • The SWNTs/HA composite coatings induced better cell proliferation, cell viability and ALP activity compared to pure HA coating and pure Ti. • The results suggested that SWNTs/HA-0.5 and SWNTs/HA-1.0 composite coating prepared in this work is acceptable in terms of mechanical property and in-vitro bioactivity. - Abstract: Single-walled carbon nanotubes/hydroxyapatite (SWNTs/HA) composite coatings were successfully fabricated by electrochemical deposition technique. Different concentrations of SWNTs were incorporated into the apatite coating by adding functionalized SWNTs into the electrolyte. Homogeneous and crack-free SWNTs/HA composite coatings were achieved and the coatings had higher crystallinity compared to pure HA coating. In addition, the highest bonding strength of the SWNTs/HA coating reached 25.7 MPa, which was nearly 70% higher than that of pure HA coating. The in-vitro cellular biocompatibility tests revealed that SWNTs/HA composite coatings exhibited higher in-vitro bioactivity than that of pure HA coating and pure titanium (Ti). It suggests that SWNTs/HA composite coating may have enormous potential applications in the field of biomaterials, especially for the metal implants

Single-walled carbon nanotubes/hydroxyapatite coatings on titanium obtained by electrochemical deposition

Energy Technology Data Exchange (ETDEWEB)

Pei, Xibo; Zeng, Yongxiang [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Prosthodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); He, Rui [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Stomatology, the Affiliated Hospital of Hangzhou Normal University, Hangzhou 310015 (China); Li, Zhongjie; Tian, Lingyang [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Prosthodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Wang, Jian, E-mail: fero@scu.edu.cn [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Prosthodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Wan, Qianbing, E-mail: pxb1024@hotmail.com [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Prosthodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Li, Xiaoyu [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Bao, Hong [Department of Stomatology, Hospital of Chengdu Office of People' s Government of Tibetan Autonomous Region, Chengdu 610000 (China)

2014-03-01

Graphical abstract: - Highlights: • The incorporation of SWNTs into the HA coating leaded to the formation of homogeneous and crack-free composite coatings. • The highest bonding strength was detected for the SWNTs/HA-0.5 composite coating (25.70 MPa). • The SWNTs/HA composite coatings induced better cell proliferation, cell viability and ALP activity compared to pure HA coating and pure Ti. • The results suggested that SWNTs/HA-0.5 and SWNTs/HA-1.0 composite coating prepared in this work is acceptable in terms of mechanical property and in-vitro bioactivity. - Abstract: Single-walled carbon nanotubes/hydroxyapatite (SWNTs/HA) composite coatings were successfully fabricated by electrochemical deposition technique. Different concentrations of SWNTs were incorporated into the apatite coating by adding functionalized SWNTs into the electrolyte. Homogeneous and crack-free SWNTs/HA composite coatings were achieved and the coatings had higher crystallinity compared to pure HA coating. In addition, the highest bonding strength of the SWNTs/HA coating reached 25.7 MPa, which was nearly 70% higher than that of pure HA coating. The in-vitro cellular biocompatibility tests revealed that SWNTs/HA composite coatings exhibited higher in-vitro bioactivity than that of pure HA coating and pure titanium (Ti). It suggests that SWNTs/HA composite coating may have enormous potential applications in the field of biomaterials, especially for the metal implants.

Carbon nanotube-based coatings on titanium

Indian Academy of Sciences (India)

Administrator

mon method is the deposition of bioactive ceramic mate- rials on the metal ... tion of nanoparticle layer, including carbon nanoparti- ... Coatings made of CNTs provide implants with .... reaches composite of CNT built into titanium oxide formed.

beta1 integrin maintains integrity of the embryonic neocortical stem cell niche.

Directory of Open Access Journals (Sweden)

Karine Loulier

2009-08-01

Full Text Available During embryogenesis, the neural stem cells (NSC of the developing cerebral cortex are located in the ventricular zone (VZ lining the cerebral ventricles. They exhibit apical and basal processes that contact the ventricular surface and the pial basement membrane, respectively. This unique architecture is important for VZ physical integrity and fate determination of NSC daughter cells. In addition, the shorter apical process is critical for interkinetic nuclear migration (INM, which enables VZ cell mitoses at the ventricular surface. Despite their importance, the mechanisms required for NSC adhesion to the ventricle are poorly understood. We have shown previously that one class of candidate adhesion molecules, laminins, are present in the ventricular region and that their integrin receptors are expressed by NSC. However, prior studies only demonstrate a role for their interaction in the attachment of the basal process to the overlying pial basement membrane. Here we use antibody-blocking and genetic experiments to reveal an additional and novel requirement for laminin/integrin interactions in apical process adhesion and NSC regulation. Transient abrogation of integrin binding and signalling using blocking antibodies to specifically target the ventricular region in utero results in abnormal INM and alterations in the orientation of NSC divisions. We found that these defects were also observed in laminin alpha2 deficient mice. More detailed analyses using a multidisciplinary approach to analyse stem cell behaviour by expression of fluorescent transgenes and multiphoton time-lapse imaging revealed that the transient embryonic disruption of laminin/integrin signalling at the VZ surface resulted in apical process detachment from the ventricular surface, dystrophic radial glia fibers, and substantial layering defects in the postnatal neocortex. Collectively, these data reveal novel roles for the laminin/integrin interaction in anchoring embryonic NSCs

Biocorrosion of TiO2 nanoparticle coating of Ti–6Al–4V in DMEM under specific in vitro conditions

International Nuclear Information System (INIS)

Höhn, Sarah; Virtanen, Sannakaisa

2015-01-01

Highlights: • Possibility to fabricate a TiO 2 NP-coating on Ti–6Al–4V by a simple spin-coating method. • The NP-coating enhances biomimetic apatite formation on the surface immersed in DMEM. • The TiO 2 coating can efficiently reduce Al release from the alloy during immersion in DMEM. • TiO 2 NP-coating makes the surface more bioactive. - Abstract: A TiO 2 nanoparticle coating was prepared on a biomedical Ti–6Al–4V alloy using “spin-coating” technique with a colloidal suspension of TiO 2 nanopowders with the aim to optimize the surface morphology (e.g., roughness) for improved biocompatibility. The influence of a TiO 2 nanoparticle (NP) coating on the corrosion behavior, metal ion release, and biomimetic apatite formation was studied in DMEM, at 37.5 °C with a continuous supply of 5% CO 2 . Electrochemical impedance spectroscopy measurements indicate a formation of a new layer on the surface of the NP-coated sample upon 28 days immersion in DMEM. Scanning electron microscopy (SEM) and X-ray spectroscopy confirm that the surface of the NP-coated Ti–6Al–4V shows a complete coverage by a Ca–phosphate layer in contrast to the non-coated Ti–6Al–4V alloy. Hence, the TiO 2 -NP coating strongly enhances biomimetic apatite formation on the alloy surface. In addition, the TiO 2 -NP coating can efficiently reduce Al-release from the alloy, for which the bare Ti–6Al–4V alloy is significant for at least 28 days of immersion in DMEM

The integrin alphav beta3 increases cellular stiffness and cytoskeletal remodeling dynamics to facilitate cancer cell invasion

Science.gov (United States)

Mierke, Claudia Tanja

2013-01-01

The process of cancer cell invasion through the extracellular matrix (ECM) of connective tissue plays a prominent role in tumor progression and is based fundamentally on biomechanics. Cancer cell invasion usually requires cell adhesion to the ECM through the cell-matrix adhesion receptors integrins. The expression of the αvβ3 integrin is increased in several tumor types and is consistently associated with increased metastasis formation in patients. The hypothesis was that the αvβ3 integrin expression increases the invasiveness of cancer cells through increased cellular stiffness, and increased cytoskeletal remodeling dynamics. Here, the invasion of cancer cells with different αvβ3 integrin expression levels into dense three-dimensional (3D) ECMs has been studied. Using a cell sorter, two subcell lines expressing either high or low amounts of αvβ3 integrins (αvβ3high or αvβ3low cells, respectively) have been isolated from parental MDA-MB-231 breast cancer cells. αvβ3high cells showed a threefold increased cell invasion compared to αvβ3low cells. Similar results were obtained for A375 melanoma, 786-O kidney and T24 bladder carcinoma cells, and cells in which the β3 integrin subunit was knocked down using specific siRNA. To investigate whether contractile forces are essential for αvβ3 integrin-mediated increased cellular stiffness and subsequently enhanced cancer cell invasion, invasion assays were performed in the presence of myosin light chain kinase inhibitor ML-7 and Rho kinase inhibitor Y27632. Indeed, cancer cell invasiveness was reduced after addition of ML-7 and Y27632 in αvβ3high cells but not in αvβ3low cells. Moreover, after addition of the contractility enhancer calyculin A, an increase in pre-stress in αvβ3low cells was observed, which enhanced cellular invasiveness. In addition, inhibition of the Src kinase, STAT3 or Rac1 strongly reduced the invasiveness of αvβ3high cells, whereas the invasiveness of β3 specific knock

The integrin alphav beta3 increases cellular stiffness and cytoskeletal remodeling dynamics to facilitate cancer cell invasion

International Nuclear Information System (INIS)

Mierke, Claudia Tanja

2013-01-01

The process of cancer cell invasion through the extracellular matrix (ECM) of connective tissue plays a prominent role in tumor progression and is based fundamentally on biomechanics. Cancer cell invasion usually requires cell adhesion to the ECM through the cell-matrix adhesion receptors integrins. The expression of the αvβ3 integrin is increased in several tumor types and is consistently associated with increased metastasis formation in patients. The hypothesis was that the αvβ3 integrin expression increases the invasiveness of cancer cells through increased cellular stiffness, and increased cytoskeletal remodeling dynamics. Here, the invasion of cancer cells with different αvβ3 integrin expression levels into dense three-dimensional (3D) ECMs has been studied. Using a cell sorter, two subcell lines expressing either high or low amounts of αvβ3 integrins (αvβ3 high or αvβ3 low cells, respectively) have been isolated from parental MDA-MB-231 breast cancer cells. αvβ3 high cells showed a threefold increased cell invasion compared to αvβ3 low cells. Similar results were obtained for A375 melanoma, 786-O kidney and T24 bladder carcinoma cells, and cells in which the β3 integrin subunit was knocked down using specific siRNA. To investigate whether contractile forces are essential for αvβ3 integrin-mediated increased cellular stiffness and subsequently enhanced cancer cell invasion, invasion assays were performed in the presence of myosin light chain kinase inhibitor ML-7 and Rho kinase inhibitor Y27632. Indeed, cancer cell invasiveness was reduced after addition of ML-7 and Y27632 in αvβ3 high cells but not in αvβ3 low cells. Moreover, after addition of the contractility enhancer calyculin A, an increase in pre-stress in αvβ3 low cells was observed, which enhanced cellular invasiveness. In addition, inhibition of the Src kinase, STAT3 or Rac1 strongly reduced the invasiveness of αvβ3 high cells, whereas the invasiveness of β3 specific knock

Role of β1-Integrin in Colorectal Cancer: Case-Control Study

Science.gov (United States)

Oh, Bo-Young; Kim, Kwang Ho; Chung, Soon Sup; Hong, Kyoung Sook

2014-01-01

Purpose In the metastatic process, interactions between circulating tumor cells (CTCs) and the extracellular matrix or surrounding cells are required. β1-Integrin may mediate these interactions. The aim of this study was to investigate whether β1-integrin is associated with the detection of CTCs in colorectal cancer. Methods We enrolled 30 patients with colorectal cancer (experimental group) and 30 patients with benign diseases (control group). Blood samples were obtained from each group, carcinoembryonic antigen (CEA) mRNA for CTCs marker and β1-integrin mRNA levels were estimated by using reverse transcription-polymerase chain reaction, and the results were compared between the two groups. In the experimental group, preoperative results were compared with postoperative results for each marker. In addition, we analyzed the correlation between the expressions of β1-integrin and CEA. Results CEA mRNA was detected more frequently in colorectal cancer patients than in control patients (P = 0.008). CEA mRNA was significantly reduced after surgery in the colorectal cancer patients (P = 0.032). β1-Integrin mRNA was detected more in colorectal cancer patients than in the patients with benign diseases (P < 0.001). In colorectal cancer patients, expression of β1-integrin mRNA was detected more for advanced-stage cancer than for early-stage cancer (P = 0.033) and was significantly decreased after surgery (P < 0.001). In addition, expression of β1-integrin mRNA was significantly associated with that of CEA mRNA in colorectal cancer patients (P = 0.001). Conclusion In conclusion, β1-integrin is a potential factor for forming a prognosis following surgical resection in colorectal cancer patients. β1-Integrin may be a candidate for use as a marker for early detection of micrometastatic tumor cells and for monitoring the therapeutic response in colorectal cancer patients. PMID:24851215

Biocompatibility of sol-gel hydroxyapatite-titania composite and bilayer coatings

Energy Technology Data Exchange (ETDEWEB)

Sidane, D., E-mail: dj.sidane@yahoo.fr [Laboratoire de Génie de l' Environnement (LGE), Faculté de Technologie, Université de Bejaia, 06000, Bejaia (Algeria); Rammal, H. [Equipe d' Accueil 4691 Biomatériaux et Inflammation en Site Osseux, SFR-CAP Santé (FED 4231), Université de Reims Champagne Ardenne, 1 Avenue du Maréchal Juin, 51100 Reims (France); Beljebbar, A. [UMR CNRS 7369, Equipe MéDIAN Biophotonique et Technologies pour la Santé, UFR de Pharmacie, SFR-CAP Santé (FED 4231), Université de Reims Champagne Ardenne, 51 rue Cognacq-Jay, 51096 Reims (France); Gangloff, S.C. [Equipe d' Accueil 4691 Biomatériaux et Inflammation en Site Osseux, SFR-CAP Santé (FED 4231), Université de Reims Champagne Ardenne, 1 Avenue du Maréchal Juin, 51100 Reims (France); Chicot, D. [FRE 3723 - LML - Laboratoire de Mécanique de Lille, Univ. Lille, 59000 Lille (France); Velard, F. [Equipe d' Accueil 4691 Biomatériaux et Inflammation en Site Osseux, SFR-CAP Santé (FED 4231), Université de Reims Champagne Ardenne, 1 Avenue du Maréchal Juin, 51100 Reims (France); Khireddine, H. [Laboratoire de Génie de l' Environnement (LGE), Faculté de Technologie, Université de Bejaia, 06000, Bejaia (Algeria); and others

2017-03-01

Titania-Hydroxyapatite (TiO{sub 2}/HAP) reinforced coatings are proposed to enhance the bioactivity and corrosion resistance of 316L stainless steel (316L SS). Herein, spin- and dip-coating sol-gel processes were investigated to construct two kinds of coatings: TiO{sub 2}/HAP composite and TiO{sub 2}/HAP bilayer. Physicochemical characterization highlighted the bioactivity response of the TiO{sub 2}/HAP composite once incubated in physiological conditions for 7 days whereas the TiO{sub 2}/HAP bilayer showed instability and dissolution. Biological analysis revealed a failure in human stem cells adhesion on TiO{sub 2}/HAP bilayer whereas on TiO{sub 2}/HAP composite the presence of polygonal shaped cells, possessing good behaviour attested a good biocompatibility of the composite coating. Finally, TiO{sub 2}/HAP composite with hardness up to 0.6 GPa and elastic modulus up to 18 GPa, showed an increased corrosion resistance of 316L SS. In conclusion, the user-friendly sol-gel processes led to bioactive TiO{sub 2}/HAP composite buildup suitable for biomedical applications. - Highlights: • 316L SS implant TiO{sub 2} reinforced HAP coatings were investigated and compared. • TiO{sub 2}/HAP composite had better structural features and biocompatible properties. • Improvement of 316L SS implants corrosion resistance. • TiO{sub 2}/HAP composite mechanical properties close to bone tissue • Low cost and desired material for hard tissue applications.

Biocompatibility of sol-gel hydroxyapatite-titania composite and bilayer coatings

International Nuclear Information System (INIS)

Sidane, D.; Rammal, H.; Beljebbar, A.; Gangloff, S.C.; Chicot, D.; Velard, F.; Khireddine, H.

2017-01-01

Titania-Hydroxyapatite (TiO 2 /HAP) reinforced coatings are proposed to enhance the bioactivity and corrosion resistance of 316L stainless steel (316L SS). Herein, spin- and dip-coating sol-gel processes were investigated to construct two kinds of coatings: TiO 2 /HAP composite and TiO 2 /HAP bilayer. Physicochemical characterization highlighted the bioactivity response of the TiO 2 /HAP composite once incubated in physiological conditions for 7 days whereas the TiO 2 /HAP bilayer showed instability and dissolution. Biological analysis revealed a failure in human stem cells adhesion on TiO 2 /HAP bilayer whereas on TiO 2 /HAP composite the presence of polygonal shaped cells, possessing good behaviour attested a good biocompatibility of the composite coating. Finally, TiO 2 /HAP composite with hardness up to 0.6 GPa and elastic modulus up to 18 GPa, showed an increased corrosion resistance of 316L SS. In conclusion, the user-friendly sol-gel processes led to bioactive TiO 2 /HAP composite buildup suitable for biomedical applications. - Highlights: • 316L SS implant TiO 2 reinforced HAP coatings were investigated and compared. • TiO 2 /HAP composite had better structural features and biocompatible properties. • Improvement of 316L SS implants corrosion resistance. • TiO 2 /HAP composite mechanical properties close to bone tissue • Low cost and desired material for hard tissue applications

Fabrication and in vitro evaluation of the collagen/hyaluronic acid PEM coating crosslinked with functionalized RGD peptide on titanium.

Science.gov (United States)

Huang, Ying; Luo, Qiaojie; Li, Xiaodong; Zhang, Feng; Zhao, Shifang

2012-02-01

Surface modification of titanium (Ti) using biomolecules has attracted much attention recently. In this study, a new strategy has been employed to construct a stable and bioactive coating on Ti. To this end, a derivative of hyaluronic acid (HA), i.e. HA-GRGDSPC-(SH), was synthesized. The disulfide-crosslinked Arg-Gly-Asp (RGD)-containing collagen/hyaluronic acid polyelectrolyte membrane (PEM) coating was then fabricated on Ti through the alternate deposition of collagen and HA-GRGDSPC-(SH) with five assembly cycles and subsequent crosslinking via converting free sulphydryl groups into disulfide linkages (RGD-CHC-Ti group). The assembly processes for PEM coating and the physicochemical properties of the coating were carefully characterized. The stability of PEM coating in phosphate-buffered saline solution could be adjusted by the crosslinking degree, while its degradation behaviors in the presence of glutathione were glutathione concentration dependent. The adhesion and proliferation of MC3T3-E1 cells were significantly enhanced in the RGD-CHC-Ti group. Up-regulated bone specific genes, enhanced alkaline phosphatase activity and osteocalcin production, the increased areas of mineralization were also observed in the RGD-CHC-Ti group. These results indicate that the strategy employed herein may function as an effective way to construct stable, RGD-containing bioactive coatings on Ti. Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Hydrogels with precisely controlled integrin activation dictate vascular patterning and permeability

Science.gov (United States)

Li, Shuoran; Nih, Lina R.; Bachman, Haylee; Fei, Peng; Li, Yilei; Nam, Eunwoo; Dimatteo, Robert; Carmichael, S. Thomas; Barker, Thomas H.; Segura, Tatiana

2017-09-01

Integrin binding to bioengineered hydrogel scaffolds is essential for tissue regrowth and regeneration, yet not all integrin binding can lead to tissue repair. Here, we show that through engineering hydrogel materials to promote α3/α5β1 integrin binding, we can promote the formation of a space-filling and mature vasculature compared with hydrogel materials that promote αvβ3 integrin binding. In vitro, α3/α5β1 scaffolds promoted endothelial cells to sprout and branch, forming organized extensive networks that eventually reached and anastomosed with neighbouring branches. In vivo, α3/α5β1 scaffolds delivering vascular endothelial growth factor (VEGF) promoted non-tortuous blood vessel formation and non-leaky blood vessels by 10 days post-stroke. In contrast, materials that promote αvβ3 integrin binding promoted endothelial sprout clumping in vitro and leaky vessels in vivo. This work shows that precisely controlled integrin activation from a biomaterial can be harnessed to direct therapeutic vessel regeneration and reduce VEGF-induced vascular permeability in vivo.

Outside-In Signal Transmission by Conformational Changes in Integrin Mac-11

Science.gov (United States)

Lefort, Craig T.; Hyun, Young-Min; Schultz, Joanne B.; Law, Foon-Yee; Waugh, Richard E.; Knauf, Philip A.; Kim, Minsoo

2010-01-01

Intracellular signals associated with or triggered by integrin ligation can control cell survival, differentiation, proliferation, and migration. Despite accumulating evidence that conformational changes regulate integrin affinity to its ligands, how integrin structure regulates signal transmission from the outside to the inside of the cell remains elusive. Using fluorescence resonance energy transfer, we addressed whether conformational changes in integrin Mac-1 are sufficient to transmit outside-in signals in human neutrophils. Mac-1 conformational activation induced by ligand occupancy or activating Ab binding, but not integrin clustering, triggered similar patterns of intracellular protein tyrosine phosphorylation, including Akt phosphorylation, and inhibited spontaneous neutrophil apoptosis, indicating that global conformational changes are critical for Mac-1-dependent outside-in signal transduction. In neutrophils and myeloid K562 cells, ligand ICAM-1 or activating Ab binding promoted switchblade-like extension of the Mac-1 extracellular domain and separation of the αM and β2 subunit cytoplasmic tails, two structural hallmarks of integrin activation. These data suggest the primacy of global conformational changes in the generation of Mac-1 outside-in signals. PMID:19864611

Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection.

Science.gov (United States)

Yun, Bing-Ling; Guan, Xiao-Lu; Liu, Yong-Zhen; Zhang, Yao; Wang, Yong-Qiang; Qi, Xiao-Le; Cui, Hong-Yu; Liu, Chang-Jun; Zhang, Yan-Ping; Gao, Hong-Lei; Gao, Li; Li, Kai; Gao, Yu-Long; Wang, Xiao-Mei

2016-07-08

Avian metapneumovirus (aMPV) fusion (F) protein mediates virus-cell membrane fusion to initiate viral infection, which requires F protein binding to its receptor(s) on the host cell surface. However, the receptor(s) for aMPV F protein is still not identified. All known subtype B aMPV (aMPV/B) F proteins contain a conserved Arg-Asp-Asp (RDD) motif, suggesting that the aMPV/B F protein may mediate membrane fusion via the binding of RDD to integrin. When blocked with integrin-specific peptides, aMPV/B F protein fusogenicity and viral replication were significantly reduced. Specifically we identified integrin αv and/or β1-mediated F protein fusogenicity and viral replication using antibody blocking, small interfering RNAs (siRNAs) knockdown, and overexpression. Additionally, overexpression of integrin αv and β1 in aMPV/B non-permissive cells conferred aMPV/B F protein binding and aMPV/B infection. When RDD was altered to RAE (Arg-Ala-Glu), aMPV/B F protein binding and fusogenic activity were profoundly impaired. These results suggest that integrin αvβ1 is a functional receptor for aMPV/B F protein-mediated membrane fusion and virus infection, which will provide new insights on the fusogenic mechanism and pathogenesis of aMPV. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection*

Science.gov (United States)

Yun, Bing-Ling; Guan, Xiao-Lu; Liu, Yong-Zhen; Zhang, Yao; Wang, Yong-Qiang; Qi, Xiao-Le; Cui, Hong-Yu; Liu, Chang-Jun; Zhang, Yan-Ping; Gao, Hong-Lei; Gao, Li; Li, Kai; Gao, Yu-Long; Wang, Xiao-Mei

2016-01-01

Avian metapneumovirus (aMPV) fusion (F) protein mediates virus-cell membrane fusion to initiate viral infection, which requires F protein binding to its receptor(s) on the host cell surface. However, the receptor(s) for aMPV F protein is still not identified. All known subtype B aMPV (aMPV/B) F proteins contain a conserved Arg-Asp-Asp (RDD) motif, suggesting that the aMPV/B F protein may mediate membrane fusion via the binding of RDD to integrin. When blocked with integrin-specific peptides, aMPV/B F protein fusogenicity and viral replication were significantly reduced. Specifically we identified integrin αv and/or β1-mediated F protein fusogenicity and viral replication using antibody blocking, small interfering RNAs (siRNAs) knockdown, and overexpression. Additionally, overexpression of integrin αv and β1 in aMPV/B non-permissive cells conferred aMPV/B F protein binding and aMPV/B infection. When RDD was altered to RAE (Arg-Ala-Glu), aMPV/B F protein binding and fusogenic activity were profoundly impaired. These results suggest that integrin αvβ1 is a functional receptor for aMPV/B F protein-mediated membrane fusion and virus infection, which will provide new insights on the fusogenic mechanism and pathogenesis of aMPV. PMID:27226547

Inhibition of αvβ3 integrin induces loss of cell directionality of oral squamous carcinoma cells (OSCC.

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Cyntia F Montenegro

Full Text Available The connective tissue formed by extracellular matrix (ECM rich in fibronectin and collagen consists a barrier that cancer cells have to overpass to reach blood vessels and then a metastatic site. Cell adhesion to fibronectin is mediated by αvβ3 and α5β1 integrins through an RGD motif present in this ECM protein, thus making these receptors key targets for cell migration studies. Here we investigated the effect of an RGD disintegrin, DisBa-01, on the migration of human fibroblasts (BJ and oral squamous cancer cells (OSCC, SCC25 on a fibronectin-rich environment. Time-lapse images were acquired on fibronectin-coated glass-bottomed dishes. Migration speed and directionality analysis indicated that OSCC cells, but not fibroblasts, showed significant decrease in both parameters in the presence of DisBa-01 (1μM and 2μM. Integrin expression levels of the α5, αv and β3 subunits were similar in both cell lines, while β1 subunit is present in lower levels on the cancer cells. Next, we examined whether the effects of DisBa-01 were related to changes in adhesion properties by using paxillin immunostaining and total internal reflection fluorescence TIRF microscopy. OSCCs in the presence of DisBa-01 showed increased adhesion sizes and number of maturing adhesion. The same parameters were analyzed usingβ3-GFP overexpressing cells and showed that β3 overexpression restored cell migration velocity and the number of maturing adhesion that were altered by DisBa-01. Surface plasmon resonance analysis showed that DisBa-01 has 100x higher affinity for αvβ3 integrin than forα5β1 integrin. In conclusion, our results suggest that the αvβ3 integrin is the main receptor involved in cell directionality and its blockage may be an interesting alternative against metastasis.

Epitopes in α8β1 and other RGD-binding integrins delineate classes of integrin-blocking antibodies and major binding loops in α subunits.

Science.gov (United States)

Nishimichi, Norihisa; Kawashima, Nagako; Yokosaki, Yasuyuki

2015-09-09

Identification of epitopes for integrin-blocking monoclonal antibodies (mAbs) has aided our understanding of structure-function relationship of integrins. We mapped epitopes of chicken anti-integrin-α8-subunit-blocking mAbs by mutational analyses, examining regions that harboured all mapped epitopes recognized by mAbs against other α-subunits in the RGD-binding-integrin subfamily. Six mAbs exhibited blocking function, and these mAbs recognized residues on the same W2:41-loop on the top-face of the β-propeller. Loop-tips sufficiently close to W2:41 (face was identified as an additional component of the epitope of one antibody, clone YZ5. Binding sequences on the two loops were conserved in virtually all mammals, and that on W3:34 was also conserved in chickens. These indicate 1) YZ5 binds both top and bottom loops, and the binding to W3:34 is by interactions to conserved residues between immunogen and host species, 2) five other blocking mAbs solely bind to W2:41 and 3) the α8 mAbs would cross-react with most mammals. Comparing with the mAbs against the other α-subunits of RGD-integrins, two classes were delineated; those binding to "W3:34 and an top-loop", and "solely W2:41", accounting for 82% of published RGD-integrin-mAbs.

Loss of the α2β1 integrin alters human papilloma virus-induced squamous carcinoma progression in vivo and in vitro.

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Thuy Tran

Full Text Available Expression of the α2β1 integrin, a receptor for collagens and laminin, is altered during tumor progression. Recent studies have linked polymorphisms in the α2 integrin gene with oral, squamous cell carcinoma (SCC. To determine the α2β1 integrin's role in SCC progression, we crossed α2-null mice with K14-HPV16 transgenic animals. Pathological progression to invasive carcinoma was evaluated in HPV-positive, α2-null (HPV/KO and HPV-positive, wild-type (HPV/WT animals. α2β1 integrin expression stimulated progression from hyperplasia and papillomatosis to dysplasia with concomitant dermal mast cell infiltration. Moreover, lymph node metastasis was decreased by 31.3% in HPV/KO, compared to HPV/WT, animals. To evaluate the integrin-specific impact on the malignant epithelium versus the microenvironment, we developed primary tumor cell lines. Although transition from dysplasia to carcinoma was unaltered during spontaneous tumor development, isolated primary HPV/KO SCC cell lines demonstrated decreased migration and invasion, compared to HPV/WT cells. When HPV/WT and HPV/KO SCC cells were orthotopically injected into WT or KO hosts, tumor α2β1 integrin expression resulted in decreased tumor latency, regardless of host integrin status. HPV/WT SCC lines failed to demonstrate a proliferative advantage in vitro, however, the HPV/WT tumors demonstrated increased growth compared to HPV/KO SCC lines in vivo. Although contributions of the integrin to the microenvironment cannot be excluded, our studies indicate that α2β1 integrin expression by HPV-transformed keratinocytes modulates SCC growth and progression.

Slurry Coating System Statement of Work and Specification

Energy Technology Data Exchange (ETDEWEB)

Chan, S. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

2017-02-06

The Slurry Coating System will be used to coat crystals with a polymer to support Lawrence Livermore National Security, LLC (LLNS) research and development at Lawrence Livermore National Laboratory (LLNL). The crystals will be suspended in water in a kettle. A polymer solution is added, temperature of the kettle is raised and aggregates of the crystals and polymer form. The slurry is heated under vacuum to drive off the solvents and slowly cooled while mixing to room temperature. The resulting aggregates are then filtered and dried. The performance characteristics and fielding constraints define a unique set of requirements for a new system. This document presents the specifications and requirements for the system.

Improved cell activity on biodegradable photopolymer scaffolds using titanate nanotube coatings

Energy Technology Data Exchange (ETDEWEB)

Beke, S., E-mail: szabolcs.beke@iit.it [Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova (Italy); Barenghi, R. [IEIIT, National Research Council (CNR), Via De Marini 6, 16149 Genova (Italy); Farkas, B.; Romano, I. [Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova (Italy); Kőrösi, L. [Department of Biotechnology, Nanophage Therapy Center, Enviroinvest Corporation, Kertváros u. 2, H-7632 Pécs (Hungary); Scaglione, S. [IEIIT, National Research Council (CNR), Via De Marini 6, 16149 Genova (Italy); Brandi, F. [Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova (Italy); Istituto Nazionale di Ottica, CNR, Via G. Moruzzi 1, 56124-Pisa (Italy)

2014-11-01

The development of bioactive materials is in the premise of tissue engineering. For several years, surface functionalization of scaffolds has been one of the most promising approaches to stimulate cellular activity and finally improve implant success. Herein, we describe the development of a bioactive composite scaffold composed of a biodegradable photopolymer scaffold and titanate nanotubes (TNTs). The biodegradable photopolymer scaffolds were fabricated by applying mask-projection excimer laser photocuring at 308 nm. TNTs were synthesized and then spin-coated on the porous scaffolds. Upon culturing fibroblast cells on scaffolds, we found that nanotubes coating affects cell viability and proliferation demonstrating that TNT coatings enhance cell growth on the scaffolds by further improving their surface topography. - Highlights: • Biodegradable scaffolds were produced by mask-assisted UV laser photocuring. • Titanate nanotube deposition was carried out without binding compounds or additives. • Titanate nanotube coatings enhanced cell viability and proliferation.

Quantitative gene-expression of the tumor angiogenesis markers vascular endothelial growth factor, integrin alphaV and integrin beta3 in human neuroendocrine tumors

DEFF Research Database (Denmark)

Oxboel, Jytte; Binderup, Tina; Knigge, Ulrich

2009-01-01

, in neuroendocrine tumors. We used quantitative real-time PCR for measuring mRNA gene-expression of vascular endothelial growth factor (VEGF), integrin alphaV, and integrin beta3, and CD34 for a group of patients with neuroendocrine tumors (n=13). Tissue from patients with colorectal cancer liver metastases (n=14...... compared to both colorectal liver metastases (p=0.10) and normal liver tissue (p=0.06). In neuroendocrine tumors, gene-expression was highly variable of VEGF (530-fold), integrin alphaV (23-fold) and integrin beta3 (106-fold). Quantitative gene-expression levels of the key angiogenesis molecules VEGF......Anti-angiogenesis treatment is a promising new therapy for cancer that recently has also been suggested for patients with neuroendocrine tumors. The aim of the present study was therefore to investigate the level of tumor angiogenesis, and thereby the molecular basis for anti-angiogenesis treatment...

Functional single-walled carbon nanotubes based on an integrin αvβ3 monoclonal antibody for highly efficient cancer cell targeting

International Nuclear Information System (INIS)

Ou Zhongmin; Wu Baoyan; Xing Da; Zhou Feifan; Wang Huiying; Tang Yonghong

2009-01-01

The application of single-walled carbon nanotubes (SWNTs) in the field of biomedicine is becoming an entirely new and exciting topic. In this study, a novel functional SWNT based on an integrin α v β 3 monoclonal antibody was developed and was used for cancer cell targeting in vitro. SWNTs were first modified by phospholipid-bearing polyethylene glycol (PL-PEG). The PL-PEG functionalized SWNTs were then conjugated with protein A. A SWNT-integrin α v β 3 monoclonal antibody system (SWNT-PEG-mAb) was thus constructed by conjugating protein A with the fluorescein labeled integrin α v β 3 monoclonal antibody. In vitro study revealed that SWNT-PEG-mAb presented a high targeting efficiency on integrin α v β 3 -positive U87MG cells with low cellular toxicity, while for integrin α v β 3 -negative MCF-7 cells, the system had a low targeting efficiency, indicating that the high targeting to U87MG cells was due to the specific integrin targeting of the monoclonal antibody. In conclusion, SWNT-PEG-mAb developed in this research is a potential candidate for cancer imaging and drug delivery in cancer targeting therapy.

Targeting ILK and β4 integrin abrogates the invasive potential of ovarian cancer

International Nuclear Information System (INIS)

Choi, Yoon Pyo; Kim, Baek Gil; Gao, Ming-Qing; Kang, Suki; Cho, Nam Hoon

2012-01-01

Highlights: ► The potential of targeting ILK and integrins for highly aggressive ovarian cancer. ► Unanticipated synergistic effect for the combination of ILK/β4 integrin. ► Combination of ILK/β4 integrin effectively inhibited the PI3K/Akt/Rac1 cascade. ► Targeting of β4 integrin/ILK had potent inhibitory effects in ovarian cancer. -- Abstract: Integrins and integrin-linked kinase (ILK) are essential to cancerous invasion because they mediate physical interactions with the extracellular matrix, and regulate oncogenic signaling pathways. The purpose of our study is to determine whether deletion of β1 and β4 integrin and ILK, alone or in combination, has antitumoral effects in ovarian cancer. Expression of β1 and β4 integrin and ILK was analyzed by immunohistochemistry in 196 ovarian cancer tissue samples. We assessed the effects of depleting these molecules with shRNAs in ovarian cancer cells by Western blot, conventional RT-PCR, cell proliferation, migration, invasion, and in vitro Rac1 activity assays, and in vivo xenograft formation assays. Overexpression of β4 integrin and ILK in human ovarian cancer specimens was found to correlate with tumor aggressiveness. Depletion of these targets efficiently suppresses ovarian cancer cell proliferation, migration, and invasion in vitro and xenograft tumor formation in vivo. We also demonstrated that single depletion of ILK or combination depletion of β4 integrin/ILK inhibits phosphorylation of downstream signaling targets, p-Ser 473 Akt and p-Thr202/Tyr204 Erk1/2, and activation of Rac1, as well as reduce expression of MMP-2 and MMP-9 and increase expression of caspase-3 in vitro. In conclusion, targeting β4 integrin combined with ILK can instigate the latent tumorigenic potential and abrogate the invasive potential in ovarian cancer.

Targeting ILK and {beta}4 integrin abrogates the invasive potential of ovarian cancer

Energy Technology Data Exchange (ETDEWEB)

Choi, Yoon Pyo; Kim, Baek Gil [BK21 Project for Medical Science, Yonsei University College of Medicine, Seoul (Korea, Republic of); Department of Pathology, Yonsei University College of Medicine, Seoul (Korea, Republic of); Gao, Ming-Qing; Kang, Suki [Department of Pathology, Yonsei University College of Medicine, Seoul (Korea, Republic of); Cho, Nam Hoon, E-mail: cho1988@yuhs.ac [BK21 Project for Medical Science, Yonsei University College of Medicine, Seoul (Korea, Republic of); Department of Pathology, Yonsei University College of Medicine, Seoul (Korea, Republic of)

2012-10-26

Highlights: Black-Right-Pointing-Pointer The potential of targeting ILK and integrins for highly aggressive ovarian cancer. Black-Right-Pointing-Pointer Unanticipated synergistic effect for the combination of ILK/{beta}4 integrin. Black-Right-Pointing-Pointer Combination of ILK/{beta}4 integrin effectively inhibited the PI3K/Akt/Rac1 cascade. Black-Right-Pointing-Pointer Targeting of {beta}4 integrin/ILK had potent inhibitory effects in ovarian cancer. -- Abstract: Integrins and integrin-linked kinase (ILK) are essential to cancerous invasion because they mediate physical interactions with the extracellular matrix, and regulate oncogenic signaling pathways. The purpose of our study is to determine whether deletion of {beta}1 and {beta}4 integrin and ILK, alone or in combination, has antitumoral effects in ovarian cancer. Expression of {beta}1 and {beta}4 integrin and ILK was analyzed by immunohistochemistry in 196 ovarian cancer tissue samples. We assessed the effects of depleting these molecules with shRNAs in ovarian cancer cells by Western blot, conventional RT-PCR, cell proliferation, migration, invasion, and in vitro Rac1 activity assays, and in vivo xenograft formation assays. Overexpression of {beta}4 integrin and ILK in human ovarian cancer specimens was found to correlate with tumor aggressiveness. Depletion of these targets efficiently suppresses ovarian cancer cell proliferation, migration, and invasion in vitro and xenograft tumor formation in vivo. We also demonstrated that single depletion of ILK or combination depletion of {beta}4 integrin/ILK inhibits phosphorylation of downstream signaling targets, p-Ser 473 Akt and p-Thr202/Tyr204 Erk1/2, and activation of Rac1, as well as reduce expression of MMP-2 and MMP-9 and increase expression of caspase-3 in vitro. In conclusion, targeting {beta}4 integrin combined with ILK can instigate the latent tumorigenic potential and abrogate the invasive potential in ovarian cancer.

Angiotensin converting enzyme (ACE and ACE2 bind integrins and ACE2 regulates integrin signalling.

Directory of Open Access Journals (Sweden)

Nicola E Clarke

Full Text Available The angiotensin converting enzymes (ACEs are the key catalytic components of the renin-angiotensin system, mediating precise regulation of blood pressure by counterbalancing the effects of each other. Inhibition of ACE has been shown to improve pathology in cardiovascular disease, whilst ACE2 is cardioprotective in the failing heart. However, the mechanisms by which ACE2 mediates its cardioprotective functions have yet to be fully elucidated. Here we demonstrate that both ACE and ACE2 bind integrin subunits, in an RGD-independent manner, and that they can act as cell adhesion substrates. We show that cellular expression of ACE2 enhanced cell adhesion. Furthermore, we present evidence that soluble ACE2 (sACE2 is capable of suppressing integrin signalling mediated by FAK. In addition, sACE2 increases the expression of Akt, thereby lowering the proportion of the signalling molecule phosphorylated Akt. These results suggest that ACE2 plays a role in cell-cell interactions, possibly acting to fine-tune integrin signalling. Hence the expression and cleavage of ACE2 at the plasma membrane may influence cell-extracellular matrix interactions and the signalling that mediates cell survival and proliferation. As such, ectodomain shedding of ACE2 may play a role in the process of pathological cardiac remodelling.

Biocorrosion of TiO{sub 2} nanoparticle coating of Ti–6Al–4V in DMEM under specific in vitro conditions

Energy Technology Data Exchange (ETDEWEB)

Höhn, Sarah, E-mail: sarah.hoehn@fau.de; Virtanen, Sannakaisa, E-mail: virtanen@ww.uni-erlangen.de

2015-02-28

Highlights: • Possibility to fabricate a TiO{sub 2} NP-coating on Ti–6Al–4V by a simple spin-coating method. • The NP-coating enhances biomimetic apatite formation on the surface immersed in DMEM. • The TiO{sub 2} coating can efficiently reduce Al release from the alloy during immersion in DMEM. • TiO{sub 2} NP-coating makes the surface more bioactive. - Abstract: A TiO{sub 2} nanoparticle coating was prepared on a biomedical Ti–6Al–4V alloy using “spin-coating” technique with a colloidal suspension of TiO{sub 2} nanopowders with the aim to optimize the surface morphology (e.g., roughness) for improved biocompatibility. The influence of a TiO{sub 2} nanoparticle (NP) coating on the corrosion behavior, metal ion release, and biomimetic apatite formation was studied in DMEM, at 37.5 °C with a continuous supply of 5% CO{sub 2}. Electrochemical impedance spectroscopy measurements indicate a formation of a new layer on the surface of the NP-coated sample upon 28 days immersion in DMEM. Scanning electron microscopy (SEM) and X-ray spectroscopy confirm that the surface of the NP-coated Ti–6Al–4V shows a complete coverage by a Ca–phosphate layer in contrast to the non-coated Ti–6Al–4V alloy. Hence, the TiO{sub 2}-NP coating strongly enhances biomimetic apatite formation on the alloy surface. In addition, the TiO{sub 2}-NP coating can efficiently reduce Al-release from the alloy, for which the bare Ti–6Al–4V alloy is significant for at least 28 days of immersion in DMEM.

Integrin α6Bβ4 inhibits colon cancer cell proliferation and c-Myc activity

International Nuclear Information System (INIS)

Dydensborg, Anders Bondo; Teller, Inga C; Groulx, Jean-François; Basora, Nuria; Paré, Fréderic; Herring, Elizabeth; Gauthier, Rémy; Jean, Dominique; Beaulieu, Jean-François

2009-01-01

Integrins are known to be important contributors to cancer progression. We have previously shown that the integrin β4 subunit is up-regulated in primary colon cancer. Its partner, the integrin α6 subunit, exists as two different mRNA splice variants, α6A and α6B, that differ in their cytoplasmic domains but evidence for distinct biological functions of these α6 splice variants is still lacking. In this work, we first analyzed the expression of integrin α6A and α6B at the protein and transcript levels in normal human colonic cells as well as colorectal adenocarcinoma cells from both primary tumors and established cell lines. Then, using forced expression experiments, we investigated the effect of α6A and α6B on the regulation of cell proliferation in a colon cancer cell line. Using variant-specific antibodies, we observed that α6A and α6B are differentially expressed both within the normal adult colonic epithelium and between normal and diseased colonic tissues. Proliferative cells located in the lower half of the glands were found to predominantly express α6A, while the differentiated and quiescent colonocytes in the upper half of the glands and surface epithelium expressed α6B. A relative decrease of α6B expression was also identified in primary colon tumors and adenocarcinoma cell lines suggesting that the α6A/α6B ratios may be linked to the proliferative status of colonic cells. Additional studies in colon cancer cells showed that experimentally restoring the α6A/α6B balance in favor of α6B caused a decrease in cellular S-phase entry and repressed the activity of c-Myc. The findings that the α6Bβ4 integrin is expressed in quiescent normal colonic cells and is significantly down-regulated in colon cancer cells relative to its α6Aβ4 counterpart are consistent with the anti-proliferative influence and inhibitory effect on c-Myc activity identified for this α6Bβ4 integrin. Taken together, these findings point out the importance of integrin

Microarc oxidation discharge types and bio properties of the coating synthesized on zirconium.

Science.gov (United States)

Cengiz, Sezgin; Azakli, Yunus; Tarakci, Mehmet; Stanciu, Lia; Gencer, Yucel

2017-08-01

This study is an attempt for gaining a better understanding on relationship between microarc oxidation (MAO) coating discharge types and bioactivity of an oxide-based coating synthesized on a Zr substrate. The discharge types and the coating growth mechanism were identified by the examination of the real cross-section image of the coating microstructure. The coating was conducted by using MAO in an electrolyte containing Na 2 SiO 3 , Ca(CH 3 COO) 2 and C 3 H 7 Na 2 O 6 P, for different durations of 2.5, 5, 15, and 30mins. The effect of the process duration on the different discharge model types (Type-A, B, and C) and bioactivity of the coatings were investigated by using X-ray Diffractometry (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy-Energy-Dispersive X-ray spectroscopy measurements (SEM-EDS) and Optical Surface Profilometry (OSP). It was found that the increasing MAO duration resulted in thicker and rougher coatings. The XRD data revealed that all the samples prepared at different process durations contained the t-ZrO 2 (tetragonal zirconia) phase. During the MAO process, non-crystalline hydroxyapatite (HA) formed, which was confirmed from the FTIR data. The surface morphology, the amount and distribution of the features of the coating surface were modified by increasing voltage. The simulated body fluid (SBF) tests showed that the more bioactive surface with more HA crystals formed owing to chemical composition and high surface roughness of the coating. The pore, crack and discharge structures played a key role in apatite nucleation and growth, and provided ingrowth of apatite into discharge channels on the coating surface. Copyright © 2017 Elsevier B.V. All rights reserved.

Design, synthesis and validation of integrin α2β1-targeted probe for microPET imaging of prostate cancer

International Nuclear Information System (INIS)

Huang, Chiun-Wei; Li, Zibo; Cai, Hancheng; Chen, Kai; Shahinian, Tony; Conti, Peter S.

2011-01-01

The ability of PET to aid in the diagnosis and management of recurrent and/or disseminated metastatic prostate cancer may be enhanced by the development of novel prognostic imaging probes. Accumulating experimental evidence indicates that overexpression of integrin α 2 β 1 may correlate with progression in human prostate cancer. In this study, 64 Cu-labeled integrin α 2 β 1 -targeted PET probes were designed and evaluated for the imaging of prostate cancer. DGEA peptides conjugated with a bifunctional chelator (BFC) were developed to image integrin α 2 β 1 expression with PET in a subcutaneous PC-3 xenograft model. The microPET images were reconstructed by a two-dimensional ordered subsets expectation maximum algorithm. The average radioactivity accumulation within a tumor or an organ was quantified from the multiple region of interest volumes. The PET tracer demonstrated prominent tumor uptake in the PC-3 xenograft (integrin α 2 β 1 -positive). The receptor specificity was confirmed in a blocking experiment. Moreover, the low tracer uptake in a CWR-22 tumor model (negative control) further confirmed the receptor specificity. The sarcophagine-conjugated DGEA peptide allows noninvasive imaging of tumor-associated α 2 β 1 expression, which may be a useful PET probe for evaluating the metastatic potential of prostate cancer. (orig.)

Absence of integrin alpha 7 causes a novel form of muscular dystrophy.

Science.gov (United States)

Mayer, U; Saher, G; Fässler, R; Bornemann, A; Echtermeyer, F; von der Mark, H; Miosge, N; Pöschl, E; von der Mark, K

1997-11-01

Integrin alpha 7 beta 1 is a specific cellular receptor for the basement membrane protein laminin-1 (refs 1,2), as well as for the laminin isoforms -2 and -4 (ref. 3). The alpha 7 subunit is expressed mainly in skeletal and cardiac muscle and has been suggested to be involved in differentiation and migration processes during myogenesis. Three cytoplasmic and two extracellular splice variants that have been described are developmentally regulated and expressed in different sites in the muscle. In adult muscle, the alpha 7A and alpha 7B subunits are concentrated in myotendinous junctions but can also be detected in neuromuscular junctions and along the sarcolemmal membrane. To study the potential involvement of alpha 7 integrin, during myogenesis and its role in muscle integrity and function, we generated a null allele of the alpha 7 gene (Itga7) in the germline of mice by homologous recombination in embryonic stem (ES) cells. Surprisingly, mice homozygous for the mutation are viable and fertile, indicating that the alpha 7 beta 1 integrin is not essential for myogenesis. However, histological analysis of skeletal muscle revealed typical symptoms of a progressive muscular dystrophy starting soon after birth, but with a distinct variability in different muscle types. The observed histopathological changes strongly indicate an impairment of function of the myotendinous junctions. These findings demonstrate that alpha 7 beta 1 integrin represents an indispensable linkage between the muscle fibre and the extracellular matrix that is independent of the dystrophin-dystroglycan complex-mediated interaction of the cytoskeleton with the muscle basement membrane.

Alpha5beta1 integrin-fibronectin interactions specify liquid to solid phase transition of 3D cellular aggregates.

Directory of Open Access Journals (Sweden)

Carlos E Caicedo-Carvajal

2010-07-01

Full Text Available Tissue organization during embryonic development and wound healing depends on the ability of cells on the one hand to exchange adhesive bonds during active rearrangement and on the other to become fixed in place as tissue homeostasis is reached. Cells achieve these contradictory tasks by regulating either cell-cell adhesive bonds, mediated by cadherins, or cell-extracellular matrix (ECM connections, regulated by integrins. Integrin alpha5beta1 and soluble fibronectin (sFN are key players in cell-ECM force generation and in ECM polymerization. Here, we explore the interplay between integrin alpha5beta1 and sFN and its influence on tissue mechanical properties and cell sorting behavior.We generated a series of cell lines varying in alpha5beta1 receptor density. We then systematically explored the effects of different sFN concentrations on aggregate biomechanical properties using tissue surface tensiometry. We found previously unreported complex behaviors including the observation that interactions between fibronectin and integrin alpha5beta1 generates biphasic tissue cohesion profiles. Specifically, we show that at constant sFn concentration, aggregate cohesion increases linearly as alpha5beta1 receptor density is increased from low to moderate levels, producing a transition from viscoelastic-liquid to pseudo viscoelastic-solid behavior. However, further increase in receptor density causes an abrupt drop in tissue cohesion and a transition back to viscoelastic-liquid properties. We propose that this may be due to depletion of sFn below a critical value in the aggregate microenvironment at high alpha5beta1 levels. We also show that differential expression of alpha5beta1 integrin can promote phase-separation between cells.The interplay between alpha5-integrin and sFn contributes significantly to tissue cohesion and, depending on their level of expression, can mediate a shift from liquid to elastic behavior. This interplay represents a tunable level

Psoriasiform skin disease in transgenic pigs with high-copy ectopic expression of human integrins α2 and β1

Science.gov (United States)

Staunstrup, Nicklas Heine; Stenderup, Karin; Mortensen, Sidsel; Primo, Maria Nascimento; Steiniche, Torben; Liu, Ying; Li, Rong; Schmidt, Mette; Purup, Stig; Dagnæs-Hansen, Frederik; Schrøder, Lisbeth Dahl; Svensson, Lars; Petersen, Thomas Kongstad; Callesen, Henrik; Bolund, Lars

2017-01-01

ABSTRACT Psoriasis is a complex human-specific disease characterized by perturbed keratinocyte proliferation and a pro-inflammatory environment in the skin. Porcine skin architecture and immunity are very similar to that in humans, rendering the pig a suitable animal model for studying the biology and treatment of psoriasis. Expression of integrins, which is normally confined to the basal layer of the epidermis, is maintained in suprabasal keratinocytes in psoriatic skin, modulating proliferation and differentiation as well as leukocyte infiltration. Here, we generated minipigs co-expressing integrins α2 and β1 in suprabasal epidermal layers. Integrin-transgenic minipigs born into the project displayed skin phenotypes that correlated with the number of inserted transgenes. Molecular analyses were in good concordance with histological observations of psoriatic hallmarks, including hypogranulosis and T-lymphocyte infiltration. These findings mark the first creation of minipigs with a psoriasiform phenotype resembling human psoriasis and demonstrate that integrin signaling plays a key role in psoriasis pathology. PMID:28679670

Simulated physiological stretch increases expression of extracellular matrix proteins in human bladder smooth muscle cells via integrin α4/αv-FAK-ERK1/2 signaling pathway.

Science.gov (United States)

Chen, Shulian; Peng, Chuandu; Wei, Xin; Luo, Deyi; Lin, Yifei; Yang, Tongxin; Jin, Xi; Gong, Lina; Li, Hong; Wang, Kunjie

2017-08-01

To investigate the effect of simulated physiological stretch on the expression of extracellular matrix (ECM) proteins and the role of integrin α4/αv, focal adhesion kinase (FAK), extracellular regulated protein kinases 1/2 (ERK1/2) in the stretch-induced ECM protein expression of human bladder smooth muscle cells (HBSMCs). HBSMCs were seeded onto silicone membrane and subjected to simulated physiological stretch at the range of 5, 10, and 15% elongation. Expression of primary ECM proteins in HBSMCs was analyzed by real-time polymerase chain reaction and Western blot. Specificity of the FAK and ERK1/2 was determined by Western blot with FAK inhibitor and ERK1/2 inhibitor (PD98059). Specificity of integrin α4 and integrin αv was determined with small interfering ribonucleic acid (siRNA) transfection. The expression of collagen I (Col1), collagen III (Col3), and fibronectin (Fn) was increased significantly under the simulated physiological stretch of 10 and 15%. Integrin α4 and αv, FAK, ERK1/2 were activated by 10% simulated physiological stretch compared with the static condition. Pretreatment of ERK1/2 inhibitor, FAK inhibitor, integrin α4 siRNA, or integrin αv siRNA reduced the stretch-induced expression of ECM proteins. And FAK inhibitor decreased the stretch-induced ERK1/2 activity and ECM protein expression. Integrin α4 siRNA or integrin αv siRNA inhibited the stretch-induced activity of FAK. Simulated physiological stretch increases the expression of ECM proteins in HBSMCs, and integrin α4/αv-FAK-ERK1/2 signaling pathway partly modulates the mechano-transducing process.

Mushroom Lectins: Specificity, Structure and Bioactivity Relevant to Human Disease

Directory of Open Access Journals (Sweden)

Mohamed Ali Abol Hassan

2015-04-01

Full Text Available Lectins are non-immunoglobulin proteins that bind diverse sugar structures with a high degree of selectivity. Lectins play crucial role in various biological processes such as cellular signaling, scavenging of glycoproteins from the circulatory system, cell–cell interactions in the immune system, differentiation and protein targeting to cellular compartments, as well as in host defence mechanisms, inflammation, and cancer. Among all the sources of lectins, plants have been most extensively studied. However, more recently fungal lectins have attracted considerable attention due to their antitumor, antiproliferative and immunomodulatory activities. Given that only 10% of mushroom species are known and have been taxonomically classified, mushrooms represent an enormous unexplored source of potentially useful and novel lectins. In this review we provide an up-to-date summary on the biochemical, molecular and structural properties of mushroom lectins, as well as their versatile applications specifically focusing on mushroom lectin bioactivity.

The membrane-cytoplasm interface of integrin alpha subunits is critical for receptor latency.

OpenAIRE

Briesewitz, R; Kern, A; Smilenov, L B; David, F S; Marcantonio, E E

1996-01-01

Localization of integrin receptors to focal contact sites occurs upon ligand binding. This activity is latent, since unoccupied integrin receptors do not localize to focal contacts. Deletion analysis has revealed that the alpha cytoplasmic domains is required for the maintenance of integrin receptor latency. Our current hypothesis for the mechanism of integrin post-ligand binding events is that there is a change in relationship of alpha and beta cytoplasmic domains, which overcomes receptor l...

LFA-1 and Mac-1 integrins bind to the serine/threonine-rich domain of thrombomodulin

Energy Technology Data Exchange (ETDEWEB)

Kawamoto, Eiji [Department of Molecular Pathobiology and Cell Adhesion Biology, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507 (Japan); Emergency and Critical Care Center, Mie University Hospital, 2-174 Edobashi, Tsu 514-8507 (Japan); Okamoto, Takayuki, E-mail: okamotot@doc.medic.mie-u.ac.jp [Department of Molecular Pathobiology and Cell Adhesion Biology, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507 (Japan); Takagi, Yoshimi [Department of Molecular Pathobiology and Cell Adhesion Biology, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507 (Japan); Honda, Goichi [Medical Affairs Department, Asahi Kasei Pharma Corporation, 1-105 Kanda Jinbo-cho, Chiyoda-ku, Tokyo 101-8101 (Japan); Suzuki, Koji [Faculty of Pharmaceutical Science, Suzuka University of Medical Science, 3500-3, Minamitamagaki-cho, Suzuka, Mie 513-8679 (Japan); Imai, Hiroshi [Emergency and Critical Care Center, Mie University Hospital, 2-174 Edobashi, Tsu 514-8507 (Japan); Shimaoka, Motomu, E-mail: shimaoka@doc.medic.mie-u.ac.jp [Department of Molecular Pathobiology and Cell Adhesion Biology, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507 (Japan)

2016-05-13

LFA-1 (αLβ2) and Mac-1 (αMβ2) integrins regulate leukocyte trafficking in health and disease by binding primarily to IgSF ligand ICAM-1 and ICAM-2 on endothelial cells. Here we have shown that the anti-coagulant molecule thrombomodulin (TM), found on the surface of endothelial cells, functions as a potentially new ligand for leukocyte integrins. We generated a recombinant extracellular domain of human TM and Fc fusion protein (TM-domains 123-Fc), and showed that pheripheral blood mononuclear cells (PBMCs) bind to TM-domains 123-Fc dependent upon integrin activation. We then demonstrated that αL integrin-blocking mAb, αM integrin-blocking mAb, and β2 integrin-blocking mAb inhibited the binding of PBMCs to TM-domains 123-Fc. Furthermore, we show that the serine/threonine-rich domain (domain 3) of TM is required for the interaction with the LFA-1 (αLβ2) and Mac-1 (αMβ2) integrins to occur on PBMCs. These results demonstrate that the LFA-1 and Mac-1 integrins on leukocytes bind to TM, thereby establishing the molecular and structural basis underlying LFA-1 and Mac-1 integrin interaction with TM on endothelial cells. In fact, integrin-TM interactions might be involved in the dynamic regulation of leukocyte adhesion with endothelial cells. - Highlights: • LFA-1 and Mac-1 integrins bind to the anti-coagulant molecule thrombomodulin. • The serine/threonine-rich domain of thrombomodulin is essential to interact with the LFA-1 and Mac-1 integrins on PBMCs. • Integrin-TM interactions might be involved in the dynamic regulation of leukocyte adhesion with endothelial cells.

Expression Profile of the Integrin Receptor Subunits in the Guinea Pig Sclera.

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Wang, Kevin K; Metlapally, Ravikanth; Wildsoet, Christine F

2017-06-01

The ocular dimensional changes in myopia reflect increased scleral remodeling, and in high myopia, loss of scleral integrity leads to biomechanical weakening and continued scleral creep. As integrins, a type of cell surface receptors, have been linked to scleral remodeling, they represent potential targets for myopia therapies. As a first step, this study aimed to characterize the integrin subunits at the messenger RNA level in the sclera of the guinea pig, a more recently added but increasingly used animal model for myopia research. Primers for α and β integrin subunits were designed using NCBI/UCSC Genome Browser and Primer3 software tools. Total RNA was extracted from normal scleral tissue and isolated cultured scleral fibroblasts, as well as liver and lung, as reference tissues, all from guinea pig. cDNA was produced by reverse transcription, PCR was used to amplify products of predetermined sizes, and products were sequenced using standard methods. Guinea pig scleral tissue expressed all known integrin alpha subunits except αD and αE. The latter integrin subunits were also not expressed by cultured guinea pig scleral fibroblasts; however, their expression was confirmed in guinea pig liver. In addition, isolated cultured fibroblasts did not express integrin subunits αL, αM, and αX. This difference between results for cultured cells and intact sclera presumably reflects the presence in the latter of additional cell types. Both guinea pig scleral tissue and isolated scleral fibroblasts expressed all known integrin beta subunits. All results were verified through sequencing. The possible contributions of integrins to scleral remodeling make them plausible targets for myopia prevention. Data from this study will help guide future ex vivo and in vitro studies directed at understanding the relationship between scleral integrins and ocular growth regulation in the guinea pig model for myopia.

The therapeutic potential of I-domain integrins.

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Brennan, Marian; Cox, Dermot

2014-01-01

Due to their role in processes central to cancer and autoimmune disease I-domain integrins are an attractive drug target. Both antibodies and small molecule antagonists have been discovered and tested in the clinic. Much of the effort has focused on αLβ2 antagonists. Maybe the most successful was the monoclonal antibody efalizumab, which was approved for the treatment of psoriasis but subsequently withdrawn from the market due to the occurrence of a serious adverse effect (progressive multifocal leukoencephalopathy). Other monoclonal antibodies were tested for the treatment of reperfusion injury, post-myocardial infarction, but failed to progress due to lack of efficacy. New potent small molecule inhibitors of αv integrins are promising reagents for treating fibrotic disease. Small molecule inhibitors targeting collagen-binding integrins have been discovered and future work will focus on identifying molecules selectively targeting each of the collagen receptors and identifying appropriate target diseases for future clinical studies.

Alternagin-C, a disintegrin-like protein from the venom of Bothrops alternatus, modulates a2ß1 integrin-mediated cell adhesion, migration and proliferation

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Selistre-de-Araujo H.S.

2005-01-01

Full Text Available The alpha2ß1 integrin is a major collagen receptor that plays an essential role in the adhesion of normal and tumor cells to the extracellular matrix. Alternagin-C (ALT-C, a disintegrin-like protein purified from the venom of the Brazilian snake Bothrops alternatus, competitively interacts with the alpha2ß1 integrin, thereby inhibiting collagen binding. When immobilized in plate wells, ALT-C supports the adhesion of fibroblasts as well as of human vein endothelial cells (HUVEC and does not detach cells previously bound to collagen I. ALT-C is a strong inducer of HUVEC proliferation in vitro. Gene expression analysis was done using an Affimetrix HU-95A probe array with probe sets of ~10,000 human genes. In human fibroblasts growing on collagen-coated plates, ALT-C up-regulates the expression of several growth factors including vascular endothelial growth factor, as well as some cell cycle control genes. Up-regulation of the vascular endothelial growth factor gene and other growth factors could explain the positive effect on HUVEC proliferation. ALT-C also strongly activates protein kinase B phosphorylation, a signaling event involved in endothelial cell survival and angiogenesis. In human neutrophils, ALT-C has a potent chemotactic effect modulated by the intracellular signaling cascade characteristic of integrin-activated pathways. Thus, ALT-C acts as a survival factor, promoting adhesion, migration and endothelial cell proliferation after binding to alpha2ß1 integrin on the cell surface. The biological activities of ALT-C may be helpful as a therapeutic strategy in tissue regeneration as well as in the design of new therapeutic agents targeting alpha2ß1 integrin.

Canine chondrodysplasia caused by a truncating mutation in collagen-binding integrin alpha subunit 10.

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Kaisa Kyöstilä

Full Text Available The skeletal dysplasias are disorders of the bone and cartilage tissues. Similarly to humans, several dog breeds have been reported to suffer from different types of genetic skeletal disorders. We have studied the molecular genetic background of an autosomal recessive chondrodysplasia that affects the Norwegian Elkhound and Karelian Bear Dog breeds. The affected dogs suffer from disproportionate short stature dwarfism of varying severity. Through a genome-wide approach, we mapped the chondrodysplasia locus to a 2-Mb region on canine chromosome 17 in nine affected and nine healthy Elkhounds (praw = 7.42×10(-6, pgenome-wide = 0.013. The associated locus contained a promising candidate gene, cartilage specific integrin alpha 10 (ITGA10, and mutation screening of its 30 exons revealed a nonsense mutation in exon 16 (c.2083C>T; p.Arg695* that segregated fully with the disease in both breeds (p = 2.5×10(-23. A 24% mutation carrier frequency was indicated in NEs and an 8% frequency in KBDs. The ITGA10 gene product, integrin receptor α10-subunit combines into a collagen-binding α10β1 integrin receptor, which is expressed in cartilage chondrocytes and mediates chondrocyte-matrix interactions during endochondral ossification. As a consequence of the nonsense mutation, the α10-protein was not detected in the affected cartilage tissue. The canine phenotype highlights the importance of the α10β1 integrin in bone growth, and the large animal model could be utilized to further delineate its specific functions. Finally, this study revealed a candidate gene for human chondrodysplasias and enabled the development of a genetic test for breeding purposes to eradicate the disease from the two dog breeds.

Synthesis and evaluation of a radioiodinated peptide probe targeting αvβ6 integrin for the detection of pancreatic ductal adenocarcinoma

International Nuclear Information System (INIS)

Ueda, Masashi; Fukushima, Takahiro; Ogawa, Kei; Kimura, Hiroyuki; Ono, Masahiro; Yamaguchi, Takashi; Ikehara, Yuzuru; Saji, Hideo

2014-01-01

Highlights: • We developed a radioiodinated peptide probe targeting αvβ6 integrin ( 123 I-IFMDV2). • 123 I-IFMDV2 had a high affinity and selectivity for αvβ6 integrin. • 123 I-IFMDV2 showed a specific binding to αvβ6 integrin in vivo. • 123 I-IFMDV2 enabled clear visualization of the αvβ6-integrin-positive tumor. - Abstract: Introduction: Pancreatic ductal adenocarcinoma (PDAC) remains a major cause of cancer-related death. Since significant upregulation of αvβ6 integrin has been reported in PDAC, this integrin is a promising target for PDAC detection. In this study, we aimed to develop a radioiodinated probe for the imaging of αvβ6 integrin-positive PDAC with single-photon emission computed tomography (SPECT). Methods: Four peptide probes were synthesized and screened by competitive and saturation binding assays using 2 PDAC cell lines (AsPC-1, αvβ6 integrin-positive; MIA PaCa-2, αvβ6 integrin-negative). The probe showing the best affinity was used to study the biodistribution assay, an in vivo blocking study, and SPECT imaging using tumor bearing mice. Autoradiography and immunohistochemical analysis were also performed. Results: Among the 4 probes examined in this study, 125 I-IFMDV2 showed the highest affinity for αvβ6 integrin expressed in AsPC-1 cells and no affinity for MIA PaCa-2 cells. The accumulation of 125 I-IFMDV2 in the AsPC-1 xenograft was 3–5 times greater than that in the MIA PaCa-2 xenograft, consistent with the expression of αvβ6 integrin in each xenograft, and confirmed by immunohistochemistry. Pretreatment with excess amounts of A20FMDV2 significantly blocked the accumulation of 125 I-IFMDV2 in the AsPC-1 xenograft, but not in the MIA PaCa-2 xenograft. Furthermore, 123 I-IFMDV2 enabled clear visualization of the AsPC-1 xenograft. Conclusion: 123 I-IFMDV2 is a potential SPECT probe for the imaging of αvβ6 integrin in PDAC

Integrins in cell migration – the actin connection

OpenAIRE

Vicente-Manzanares, Miguel; Choi, Colin Kiwon; Horwitz, Alan Rick

2008-01-01

The connection between integrins and actin is driving the field of cell migration in new directions. Integrins and actin are coupled through a physical linkage, which provides traction for migration. Recent studies show the importance of this linkage in regulating adhesion organization and development. Actin polymerization orchestrates adhesion assembly near the leading edge of a migrating cell, and the dynamic cross-linking of actin filaments promotes adhesion maturat...

The Bioactivity and Photocatalytic Properties of Titania Nanotube Coatings Produced with the Use of the Low-Potential Anodization of Ti6Al4V Alloy Surface

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Radtke, Aleksandra; Kozak, Wiesław; Sadowska, Beata; Więckowska-Szakiel, Marzena; Szubka, Magdalena; Talik, Ewa; Pleth Nielsen, Lars; Piszczek, Piotr

2017-01-01

Titania nanotube (TNT) coatings were produced using low-potential anodic oxidation of Ti6Al4V substrates in the potential range 3–20 V. They were analysed by X-ray diffraction (XRD), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM). The wettability was estimated by measuring the contact angle when applying water droplets. The bioactivity of the TNT coatings was established on the basis of the biointegration assay (L929 murine fibroblasts adhesion and proliferation) and antibacterial tests against Staphylococcus aureus (ATCC 29213). The photocatalytic efficiency of the TNT films was studied by the degradation of methylene blue under UV irradiation. Among the studied coatings, the TiO2 nanotubes obtained with the use of 5 V potential (TNT5) were found to be the most appropriate for medical applications. The TNT5 sample possessed antibiofilm properties without enriching it by additional antimicrobial agent. Furthermore, it was characterized by optimal biocompatibility, performing better than pure Ti6Al4V alloy. Moreover, the same sample was the most photocatalytically active and exhibited the potential for the sterilization of implants with the use of UV light and for other environmental applications. PMID:28933732

The Bioactivity and Photocatalytic Properties of Titania Nanotube Coatings Produced with the Use of the Low-Potential Anodization of Ti6Al4V Alloy Surface.

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Radtke, Aleksandra; Topolski, Adrian; Jędrzejewski, Tomasz; Kozak, Wiesław; Sadowska, Beata; Więckowska-Szakiel, Marzena; Szubka, Magdalena; Talik, Ewa; Pleth Nielsen, Lars; Piszczek, Piotr

2017-07-26

Titania nanotube (TNT) coatings were produced using low-potential anodic oxidation of Ti6Al4V substrates in the potential range 3-20 V. They were analysed by X-ray diffraction (XRD), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM). The wettability was estimated by measuring the contact angle when applying water droplets. The bioactivity of the TNT coatings was established on the basis of the biointegration assay (L929 murine fibroblasts adhesion and proliferation) and antibacterial tests against Staphylococcus aureus (ATCC 29213). The photocatalytic efficiency of the TNT films was studied by the degradation of methylene blue under UV irradiation. Among the studied coatings, the TiO₂ nanotubes obtained with the use of 5 V potential (TNT5) were found to be the most appropriate for medical applications. The TNT5 sample possessed antibiofilm properties without enriching it by additional antimicrobial agent. Furthermore, it was characterized by optimal biocompatibility, performing better than pure Ti6Al4V alloy. Moreover, the same sample was the most photocatalytically active and exhibited the potential for the sterilization of implants with the use of UV light and for other environmental applications.

Hypergravity Stimulates the Extracellular Matrix/Integrin-Signaling Axis and Proliferation in Primary Osteoblasts

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Parra, M.; Vercoutere, W.; Roden, C.; Banerjee, I.; Krauser, W.; Holton, E.; Searby, N.; Globus, R.; Almeida, E.

2003-01-01

We set out to determine the molecular mechanisms involved in the proliferative response of primary rat osteoblasts to mechanical stimulation using cell culture centrifugation as a model for hypergravity. We hypothesized that this proliferative response is mediated by specific integrin/Extracellular Matrix (ECM) interactions. To investigate this question we developed a cell culture centrifuge and an automated system that performs cell fixation during hypergravity loading. We generated expression vectors for various focal adhesion and cytoskeletal proteins fused to GFP or dsRed and visualized these structures in transfected (or infected) osteoblasts. The actin cytoskeleton was also visualized using rhodamine-phalloidin staining and Focal Adhesion Kinase (FAK) levels were assessed biochemically. We observed that a 24 hour exposure to 50-g stimulated proliferation compared to the 1-g control when cells were plated on fibronectin, collagen Type I , and collagen Type IV, but not on uncoated tissue culture plastic surfaces. This proliferative response was greatest for osteoblasts grown on fibronectin (2-fold increase over 1-g control) and collagen Type I (1.4 fold increase over 1-g control), suggesting that specific matrices and integrins are involved in the signaling pathways required for proliferation. Exposing osteoblasts grown on different matrices to 10-g or 25-g showed that effects on proliferation depended on both matrix type and loading level. We found that osteoblasts exposed to a short pulse of hypergravity during adhesion spread further and had more GFP-FAK containing focal adhesions compared to their 1-g controls. While overall levels of FAK did not change, more FAK was in the active (phosphorylated) form under hypergravity than in the 1-g controls. Cytoskeletal F-actin organization into filaments was also more prominent after brief exposures to hypergravity during the first five minutes of adhesion. These results suggest that specific integrins sense

Integrins Regulate Apical Constriction via Microtubule Stabilization in the Drosophila Eye Disc Epithelium

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Vilaiwan M. Fernandes

2014-12-01

Full Text Available During morphogenesis, extracellular signals trigger actomyosin contractility in subpopulations of cells to coordinate changes in cell shape. To illuminate the link between signaling-mediated tissue patterning and cytoskeletal remodeling, we study the progression of the morphogenetic furrow (MF, the wave of apical constriction that traverses the Drosophila eye imaginal disc preceding photoreceptor neurogenesis. Apical constriction depends on actomyosin contractility downstream of the Hedgehog (Hh and bone morphogenetic protein (BMP pathways. We identify a role for integrin adhesion receptors in MF progression. We show that Hh and BMP regulate integrin expression, the loss of which disrupts apical constriction and slows furrow progression; conversely, elevated integrins accelerate furrow progression. We present evidence that integrins regulate MF progression by promoting microtubule stabilization, since reducing microtubule stability rescues integrin-mediated furrow acceleration. Thus, integrins act as a genetic link between tissue-level signaling events and morphological change at the cellular level, leading to morphogenesis and neurogenesis in the eye.

Transmission of integrin β7 transmembrane domain topology enables gut lymphoid tissue development.

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Sun, Hao; Lagarrigue, Frederic; Gingras, Alexandre R; Fan, Zhichao; Ley, Klaus; Ginsberg, Mark H

2018-04-02

Integrin activation regulates adhesion, extracellular matrix assembly, and cell migration, thereby playing an indispensable role in development and in many pathological processes. A proline mutation in the central integrin β3 transmembrane domain (TMD) creates a flexible kink that uncouples the topology of the inner half of the TMD from the outer half. In this study, using leukocyte integrin α4β7, which enables development of gut-associated lymphoid tissue (GALT), we examined the biological effect of such a proline mutation and report that it impairs agonist-induced talin-mediated activation of integrin α4β7, thereby inhibiting rolling lymphocyte arrest, a key step in transmigration. Furthermore, the α4β7(L721P) mutation blocks lymphocyte homing to and development of the GALT. These studies show that impairing the ability of an integrin β TMD to transmit talin-induced TMD topology inhibits agonist-induced physiological integrin activation and biological function in development. © 2018 Sun et al.

Rapid Obtaining of Nano-Hydroxyapatite Bioactive Films on NiTi Shape Memory Alloy by Electrodeposition Process

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Lobo, A. O.; Otubo, J.; Matsushima, J. T.; Corat, E. J.

2011-07-01

Nano-hydroxyapatite (n-HA) crystalline films have been developed in this study by electrodeposition method on NiTi shape memory alloy (SMA). The electrodeposition of the n-HA films was carried out using 0.042 mol/L Ca(NO3)2 · 4H2O + 0.025 mol/L (NH4) · 2HPO4 electrolytes by applying a constant potential of -2.0 V for 120 min and keeping the solution temperature at 70 °C. The characterization of n-HA films is of special importance since bioactive properties related to n-HA have been directly identified with its specific composition and crystalline structure. AFM, XRD, EDX, FEG-SEM and Raman spectroscopy shows a homogeneous film, with high crystallinity, special composition, and bioactivity properties (Ca/P = 1.93) of n-HA on NiTi SMA surfaces. The n-HA coating with special structure would benefit the use of NiTi alloy in orthopedic applications.

ADAM disintegrin-like domain recognition by the lymphocyte integrins α4β1 and α4β7

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Bridges, Lance C.; Sheppard, Dean; Bowditch, Ron D.

2004-01-01

The ADAM (a disintegrin and metalloprotease) family of proteins possess both proteolytic and adhesive domains. We have established previously that the disintegrin domain of ADAM28, an ADAM expressed by human lymphocytes, is recognized by the integrin α4β1. The present study characterizes the integrin binding properties of the disintegrin-like domains of human ADAM7, ADAM28 and ADAM33 with the integrins α4β1, α4β7 and α9β1. Cell-adhesion assays demonstrated that, similar to ADAM28, the ADAM7 disintegrin domain supported α4β1-dependent Jurkat cell adhesion, whereas the ADAM33 disintegrin domain did not. The lymphocyte integrin α4β7 was also found to recognize both disintegrin domains of ADAM7 and ADAM28, but not of ADAM33. This is the first demonstration that mammalian disintegrins are capable of interacting with α4β7. All three disintegrin domains supported α9β1-dependent cell adhesion. Recognition by both α4β1 and α4β7 of ADAM7 and ADAM28 was activation-dependent, requiring either the presence of Mn2+ or an activating monoclonal antibody for cell attachment. Charge-to-alanine mutagenesis experiments revealed that the same residues within an individual ADAM disintegrin domain function in recognizing multiple integrins. However, the residues within a specific region of each ADAM disintegrin-like domain required for integrin binding were distinct. These results establish that ADAM7 and ADAM28 are recognized by the leucocyte integrins α4β1, α4β7 and α9β1. ADAM33 exclusively supported only α9β1-dependent adhesion. PMID:15504110

Engineered matrix coatings to modulate the adhesion of CD133+ human hematopoietic progenitor cells.

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Franke, Katja; Pompe, Tilo; Bornhäuser, Martin; Werner, Carsten

2007-02-01

Interactions of hematopoietic progenitor cells (HPC) with their local microenvironments in the bone marrow are thought to control homing, differentiation, and self-renewal of the cells. To dissect the role of extracellular matrix (ECM) components of the niche microenvironment, a set of well-defined ECM coatings including fibronectin, heparin, heparan sulphate, hyaluronic acid, tropocollagen I, and co-fibrils of collagen I with heparin or hyaluronic acid was prepared and analysed with respect to the attachment of human CD133+ HPC in vitro. The extension of the adhesion areas of individual cells as well as the fraction of adherent cells were assessed by reflection interference contrast microscopy (RICM). Intense cell-matrix interactions were found on surfaces coated with fibronectin, heparin, heparan sulphate, and on the collagen I based co-fibrils. Insignificant adhesion was found for tropocollagen I and hyaluronic acid. The strongest adhesion of HPC was observed on fibronectin with contact areas of about 7 microm(2). Interaction of HPC with coatings consisting of heparin, heparan sulphate, and co-fibrils result in small circular shaped contact zones of 3 microm(2) pointing to another, less efficient, adhesion mechanism. Analysing the specificity of cell-matrix interaction by antibody blocking experiments suggests an integrin(alpha(5)beta(1))-specific adhesion on fibronectin, while adhesion on heparin was shown to be mediated by selectins (CD62L). Taken together, our data provide a basis for the design of advanced culture carriers supporting site-specific proliferation or differentiation of HPC.

A study of strontium doped calcium phosphate coatings on AZ31

International Nuclear Information System (INIS)

Singh, Satish S.; Roy, Abhijit; Lee, Boeun E.; Ohodnicki, John; Loghmanian, Autrine; Banerjee, Ipsita; Kumta, Prashant N.

2014-01-01

Calcium phosphate (CaP) coatings have been studied to tailor the uncontrolled non-uniform corrosion of Mg based alloys while simultaneously enhancing bioactivity. The use of immersion techniques to deposit CaP coatings is attractive due to the ability of the approach to coat complex structures. In the current study, AZ31 substrates were subjected to various pretreatment conditions prior to depositing Sr 2+ doped and undoped CaP coatings. It was hypothesized that the bioactivity and corrosion protection of CaP coatings could be improved by doping with Sr 2+ . Heat treatment to elevated temperatures resulted in the diffusion of alloying elements, Mg and Zn, into the pretreated layer. Sr 2+ doped and undoped CaP coatings formed on the pretreated substrates consisted of biphasic mixtures of β-tricalcium phosphate (β-TCP) and hydroxyapatite (HA). Electrochemical corrosion experiments indicated that the extent of Sr 2+ doping and pretreatment both influenced the corrosion protection. Cytotoxicity was evaluated with MC3T3-E1 mouse preosteoblasts and human mesenchymal stem cells (hMSCs). For both cell types, proliferation decreased upon increasing the Sr 2+ concentration. However, both osteogenic gene and protein expression significantly increased upon increasing Sr 2+ concentration. These results suggest that Sr 2+ doped coatings are capable of promoting osteogenic differentiation on degradable Mg alloys, while also enhancing corrosion protection, in comparison to undoped CaP coatings

Acute Podocyte Vascular Endothelial Growth Factor (VEGF-A) Knockdown Disrupts alphaVbeta3 Integrin Signaling in the Glomerulus

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Veron, Delma; Villegas, Guillermo; Aggarwal, Pardeep Kumar; Bertuccio, Claudia; Jimenez, Juan; Velazquez, Heino; Reidy, Kimberly; Abrahamson, Dale R.; Moeckel, Gilbert; Kashgarian, Michael; Tufro, Alda

2012-01-01

Podocyte or endothelial cell VEGF-A knockout causes thrombotic microangiopathy in adult mice. To study the mechanism involved in acute and local injury caused by low podocyte VEGF-A we developed an inducible, podocyte-specific VEGF-A knockdown mouse, and we generated an immortalized podocyte cell line (VEGFKD) that downregulates VEGF-A upon doxycycline exposure. Tet-O-siVEGF:podocin-rtTA mice express VEGF shRNA in podocytes in a doxycycline-regulated manner, decreasing VEGF-A mRNA and VEGF-A protein levels in isolated glomeruli to ∼20% of non-induced controls and urine VEGF-A to ∼30% of control values a week after doxycycline induction. Induced tet-O-siVEGF:podocin-rtTA mice developed acute renal failure and proteinuria, associated with mesangiolysis and microaneurisms. Glomerular ultrastructure revealed endothelial cell swelling, GBM lamination and podocyte effacement. VEGF knockdown decreased podocyte fibronectin and glomerular endothelial alphaVbeta3 integrin in vivo. VEGF receptor-2 (VEGFR2) interacts with beta3 integrin and neuropilin-1 in the kidney in vivo and in VEGFKD podocytes. Podocyte VEGF knockdown disrupts alphaVbeta3 integrin activation in glomeruli, detected by WOW1-Fab. VEGF silencing in cultured VEGFKD podocytes downregulates fibronectin and disrupts alphaVbeta3 integrin activation cell-autonomously. Collectively, these studies indicate that podocyte VEGF-A regulates alphaVbeta3 integrin signaling in the glomerulus, and that podocyte VEGF knockdown disrupts alphaVbeta3 integrin activity via decreased VEGFR2 signaling, thereby damaging the three layers of the glomerular filtration barrier, causing proteinuria and acute renal failure. PMID:22808199

Variation in one residue associated with the metal ion-dependent adhesion site regulates αIIbβ3 integrin ligand binding affinity.

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Joel Raborn

Full Text Available The Asp of the RGD motif of the ligand coordinates with the β I domain metal ion dependent adhesion site (MIDAS divalent cation, emphasizing the importance of the MIDAS in ligand binding. There appears to be two distinct groups of integrins that differ in their ligand binding affinity and adhesion ability. These differences may be due to a specific residue associated with the MIDAS, particularly the β3 residue Ala(252 and corresponding Ala in the β1 integrin compared to the analogous Asp residue in the β2 and β7 integrins. Interestingly, mutations in the adjacent to MIDAS (ADMIDAS of integrins α4β7 and αLβ2 increased the binding and adhesion abilities compared to the wild-type, while the same mutations in the α2β1, α5β1, αVβ3, and αIIbβ3 integrins demonstrated decreased ligand binding and adhesion. We introduced a mutation in the αIIbβ3 to convert this MIDAS associated Ala(252 to Asp. By combination of this mutant with mutations of one or two ADMIDAS residues, we studied the effects of this residue on ligand binding and adhesion. Then, we performed molecular dynamics simulations on the wild-type and mutant αIIbβ3 integrin β I domains, and investigated the dynamics of metal ion binding sites in different integrin-RGD complexes. We found that the tendency of calculated binding free energies was in excellent agreement with the experimental results, suggesting that the variation in this MIDAS associated residue accounts for the differences in ligand binding and adhesion among different integrins, and it accounts for the conflicting results of ADMIDAS mutations within different integrins. This study sheds more light on the role of the MIDAS associated residue pertaining to ligand binding and adhesion and suggests that this residue may play a pivotal role in integrin-mediated cell rolling and firm adhesion.

α2 Integrin, extracellular matrix metalloproteinase inducer, and matrix metalloproteinase-3 act sequentially to induce differentiation of mouse embryonic stem cells into odontoblast-like cells

International Nuclear Information System (INIS)

Ozeki, Nobuaki; Kawai, Rie; Hase, Naoko; Hiyama, Taiki; Yamaguchi, Hideyuki; Kondo, Ayami; Nakata, Kazuhiko; Mogi, Makio

2015-01-01

We previously reported that interleukin 1β acts via matrix metalloproteinase (MMP)-3 to regulate cell proliferation and suppress apoptosis in α2 integrin-positive odontoblast-like cells differentiated from mouse embryonic stem (ES) cells. Here we characterize the signal cascade underpinning odontoblastic differentiation in mouse ES cells. The expression of α2 integrin, extracellular matrix metalloproteinase inducer (Emmprin), and MMP-3 mRNA and protein were all potently increased during odontoblastic differentiation. Small interfering RNA (siRNA) disruption of the expression of these effectors potently suppressed the expression of the odontoblastic biomarkers dentin sialophosphoprotein, dentin matrix protein-1 and alkaline phosphatase, and blocked odontoblast calcification. Our siRNA, western blot and blocking antibody analyses revealed a unique sequential cascade involving α2 integrin, Emmprin and MMP-3 that drives ES cell differentiation into odontoblasts. This cascade requires the interaction between α2 integrin and Emmprin and is potentiated by exogenous MMP-3. Finally, although odontoblast-like cells potently express α2, α6, αV, β1, and β3, integrins, we confirmed that β1 integrin acts as the trigger for ES cell differentiation, apparently in complex with α2 integrin. These results demonstrate a unique and unanticipated role for an α2 integrin-, Emmprin-, and MMP-3-mediated signaling cascade in driving mouse ES cell differentiation into odontoblast-like cells. - Highlights: • Odontoblast differentiation requires activation of α2 integrin, Emmprin and MMP-3. • α2 integrin, Emmprin and MMP-3 form a sequential signaling cascade. • β1 integrin acts a specific trigger for odontoblast differentiation. • The role of these effectors is highly novel and unanticipated

α2 Integrin, extracellular matrix metalloproteinase inducer, and matrix metalloproteinase-3 act sequentially to induce differentiation of mouse embryonic stem cells into odontoblast-like cells

Energy Technology Data Exchange (ETDEWEB)

Ozeki, Nobuaki; Kawai, Rie; Hase, Naoko; Hiyama, Taiki; Yamaguchi, Hideyuki [Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi 464-8651 (Japan); Kondo, Ayami [Department of Medicinal Biochemistry, School of Pharmacy, Aichi Gakuin University, Nagoya 464-8650 (Japan); Nakata, Kazuhiko [Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi 464-8651 (Japan); Mogi, Makio, E-mail: makio@dpc.agu.ac.jp [Department of Medicinal Biochemistry, School of Pharmacy, Aichi Gakuin University, Nagoya 464-8650 (Japan)

2015-02-01

We previously reported that interleukin 1β acts via matrix metalloproteinase (MMP)-3 to regulate cell proliferation and suppress apoptosis in α2 integrin-positive odontoblast-like cells differentiated from mouse embryonic stem (ES) cells. Here we characterize the signal cascade underpinning odontoblastic differentiation in mouse ES cells. The expression of α2 integrin, extracellular matrix metalloproteinase inducer (Emmprin), and MMP-3 mRNA and protein were all potently increased during odontoblastic differentiation. Small interfering RNA (siRNA) disruption of the expression of these effectors potently suppressed the expression of the odontoblastic biomarkers dentin sialophosphoprotein, dentin matrix protein-1 and alkaline phosphatase, and blocked odontoblast calcification. Our siRNA, western blot and blocking antibody analyses revealed a unique sequential cascade involving α2 integrin, Emmprin and MMP-3 that drives ES cell differentiation into odontoblasts. This cascade requires the interaction between α2 integrin and Emmprin and is potentiated by exogenous MMP-3. Finally, although odontoblast-like cells potently express α2, α6, αV, β1, and β3, integrins, we confirmed that β1 integrin acts as the trigger for ES cell differentiation, apparently in complex with α2 integrin. These results demonstrate a unique and unanticipated role for an α2 integrin-, Emmprin-, and MMP-3-mediated signaling cascade in driving mouse ES cell differentiation into odontoblast-like cells. - Highlights: • Odontoblast differentiation requires activation of α2 integrin, Emmprin and MMP-3. • α2 integrin, Emmprin and MMP-3 form a sequential signaling cascade. • β1 integrin acts a specific trigger for odontoblast differentiation. • The role of these effectors is highly novel and unanticipated.

Alterations in integrin expression modulates invasion of pancreatic cancer cells.

LENUS (Irish Health Repository)

Walsh, Naomi

2009-01-01

BACKGROUND: Factors mediating the invasion of pancreatic cancer cells through the extracellular matrix (ECM) are not fully understood. METHODS: In this study, sub-populations of the human pancreatic cancer cell line, MiaPaCa-2 were established which displayed differences in invasion, adhesion, anoikis, anchorage-independent growth and integrin expression. RESULTS: Clone #3 displayed higher invasion with less adhesion, while Clone #8 was less invasive with increased adhesion to ECM proteins compared to MiaPaCa-2. Clone #8 was more sensitive to anoikis than Clone #3 and MiaPaCa-2, and displayed low colony-forming efficiency in an anchorage-independent growth assay. Integrins beta 1, alpha 5 and alpha 6 were over-expressed in Clone #8. Using small interfering RNA (siRNA), integrin beta1 knockdown in Clone #8 cells increased invasion through matrigel and fibronectin, increased motility, decreased adhesion and anoikis. Integrin alpha 5 and alpha 6 knockdown also resulted in increased motility, invasion through matrigel and decreased adhesion. CONCLUSION: Our results suggest that altered expression of integrins interacting with different extracellular matrixes may play a significant role in suppressing the aggressive invasive phenotype. Analysis of these clonal populations of MiaPaCa-2 provides a model for investigations into the invasive properties of pancreatic carcinoma.

Betal-integrins in the primary cilium of MDCK cells potentiate fibronectin-induced Ca2+ signalling

DEFF Research Database (Denmark)

Prætorius, Helle; Prætorius, Jeppe; Nielsen, Søren

2004-01-01

observed that β1-integrin, α3-integrin, and perhaps α5-integrin were localized to the primary cilium of MDCK cells by combining lectin and immunofluorescence confocal microscopy. β1-Integrin was also colocalized with tubulin to the primary cilia of the rat renal collecting ducts, as well as to the cilia...

Neocellularization and neovascularization of nanosized bioactive glass-coated decellularized trabecular bone scaffolds

KAUST Repository

Gerhardt, Lutz Christian; Widdows, Kate L.; Erol, Melek M.; Nandakumar, Anandkumar; Roqan, Iman S.; Ansari, Tahera I.; Boccaccini, Aldo R.

2012-01-01

amount of vascular endothelial growth factor (VEGF) secreted by human fibroblasts grown on n-BG coatings (0-1.245 mg/cm 2), decellularized trabecular bone samples (porosity: 43-81%) were coated with n-BG particles. Grown on n-BG particles at a coating

Oncofetal Chondroitin Sulfate Glycosaminoglycans are Key Players in Integrin Signaling and Tumor Cell Motility

Science.gov (United States)

Clausen, Thomas Mandel; Pereira, Marina Ayres; Al Nakouzi, Nader; Oo, Htoo Zarni; Agerbæk, Mette Ø; Lee, Sherry; Ørum-Madsen, Maj Sofie; Christensen, Anders Riis; El-Naggar, Amal; Grandgenett, Paul M.; Grem, Jean L.; Hollingsworth, Michael A.; Holst, Peter J.; Theander, Thor; Sorensen, Poul H.; Daugaard, Mads; Salanti, Ali

2016-01-01

Many tumors express proteoglycans modified with oncofetal chondroitin sulfate glycosaminoglycan chains (ofCS), which are normally restricted to the placenta. However, the role of ofCS in cancer is largely unknown. The function of ofCS in cancer was analyzed using the recombinant ofCS-binding VAR2CSA protein (rVAR2) derived from the malaria parasite, Plasmodium falciparum. We demonstrate that ofCS plays a key role in tumor cell motility by affecting canonical integrin signaling pathways. Binding of rVAR2 to tumor cells inhibited the interaction of cells with extracellular matrix (ECM) components, which correlated with decreased phosphorylation of Src kinase. Moreover, rVAR2 binding decreased migration, invasion and anchorage-independent growth of tumor cells in vitro. Mass spectrometry of ofCS-modified proteoglycan complexes affinity purified from tumor cell lines on rVAR2 columns, revealed an overrepresentation of proteins involved in cell motility and integrin signaling, such as integrin β1 (ITGB1) and integrin α4 (ITGA4). Saturating concentrations of rVAR2 inhibited downstream integrin signaling, which was mimicked by knockdown of the core CS synthesis enzymes Beta-1,3-Glucuronyltransferase 1 (B3GAT1) and Chondroitin Sulfate N-Acetylgalactosaminyltransferase 1 (CSGALNACT1). The ofCS modification was highly expressed in both human and murine metastatic lesions in situ and pre-incubation or early intravenous treatment of tumor cells with rVAR2 inhibited seeding and spreading of tumor cells in mice. This was associated with a significant increase in survival of the animals. These data functionally link ofCS modifications with cancer cell motility and further highlights ofCS as a novel therapeutic cancer target. Implications The cancer specific expression of oncofetal chondroitin sulfate aids in metastatic phenotypes and is a candidate target for therapy. PMID:27655130

Direct Thy-1/alphaVbeta3 integrin interaction mediates neuron to astrocyte communication.

Science.gov (United States)

Hermosilla, Tamara; Muñoz, Daniel; Herrera-Molina, Rodrigo; Valdivia, Alejandra; Muñoz, Nicolás; Nham, Sang-Uk; Schneider, Pascal; Burridge, Keith; Quest, Andrew F G; Leyton, Lisette

2008-06-01

Thy-1 is an abundant neuronal glycoprotein of poorly defined function. We recently provided evidence indicating that Thy-1 clusters a beta3-containing integrin in astrocytes to induce tyrosine phosphorylation, RhoA activation and the formation of focal adhesions and stress fibers. To date, the alpha subunit partner of beta3 integrin in DI TNC1 astrocytes is unknown. Similarly, the ability of neuronal, membrane-bound Thy-1 to trigger astrocyte signaling via integrin engagement remains speculation. Here, evidence that alphav forms an alphavbeta3 heterodimer in DI TNC1 astrocytes was obtained. In neuron-astrocyte association assays, the presence of either anti-alphav or anti-beta3 integrin antibodies reduced cell-cell interaction demonstrating the requirement of both integrin subunits for this association. Moreover, anti-Thy-1 antibodies blocked stimulation of astrocytes by neurons but not the binding of these two cell types. Thus, neuron-astrocyte association involved binding between molecular components in addition to the Thy-1-integrin; however, the signaling events leading to focal adhesion formation in astrocytes depended exclusively on the latter interaction. Additionally, wild-type (RLD) but not mutated (RLE) Thy-1 was shown to directly interact with alphavbeta3 integrin by Surface Plasmon Resonance analysis. This interaction was promoted by divalent cations and was species-independent. Together, these results demonstrate that the alphavbeta3 integrin heterodimer interacts directly with Thy-1 present on neuronal cells to stimulate astrocytes.

Genetic analysis of beta1 integrin "activation motifs" in mice

DEFF Research Database (Denmark)

Czuchra, Aleksandra; Meyer, Hannelore; Legate, Kyle R

2006-01-01

-null phenotype in vivo. Surprisingly, neither the substitution of the tyrosines with phenylalanine nor the aspartic acid with alanine resulted in an obvious defect. These data suggest that the NPXY motifs of the beta1 integrin tail are essential for beta1 integrin function, whereas tyrosine phosphorylation...

Integrins and small GTPases as modulators of phagocytosis.

Science.gov (United States)

Sayedyahossein, Samar; Dagnino, Lina

2013-01-01

Phagocytosis is the mechanism whereby cells engulf large particles. This process has long been recognized as a critical component of the innate immune response, which constitutes the organism's defense against microorganisms. In addition, phagocytic internalization of apoptotic cells or cell fragments plays important roles in tissue homeostasis and remodeling. Phagocytosis requires target interactions with receptors on the plasma membrane of the phagocytic cell. Integrins have been identified as important mediators of particle clearance, in addition to their well-established roles in cell adhesion, migration and mechanotransduction. Indeed, these ubiquitously expressed proteins impart phagocytic capacity to epithelial, endothelial and mesenchymal cell types. The importance of integrins in particle internalization is emphasized by the ability of microbial and viral pathogens to exploit their signaling pathways to invade host cells, and by the wide variety of disorders that arise from abnormalities in integrin-dependent phagocytic uptake. Copyright © 2013 Elsevier Inc. All rights reserved.

Wear performance of laser processed tantalum coatings

Energy Technology Data Exchange (ETDEWEB)

Dittrick, Stanley; Balla, Vamsi Krishna; Bose, Susmita; Bandyopadhyay, Amit, E-mail: amitband@wsu.edu

2011-12-01

This first generation investigation evaluates the in vitro tribological performance of laser-processed Ta coatings on Ti for load-bearing implant applications. Linear reciprocating wear tests in simulated body fluid showed one order of magnitude less wear rate, of the order of 10{sup -4} mm{sup 3}(N.m){sup -1}, for Ta coatings compared to Ti. Our results demonstrate that Ta coatings can potentially minimize the early-stage bone-implant interface micro-motion induced wear debris generation due to their excellent bioactivity comparable to that of hydroxyapatite (HA), high wear resistance and toughness compared to popular HA coatings. Highlights: {yields} In vitro wear performance of laser processed Ta coatings on Ti was evaluated. {yields} Wear tests in SBF showed one order of magnitude less wear for Ta coatings than Ti. {yields} Ta coatings can minimize early-stage micro-motion induced wear debris generation.

Assessing activation of hepatic stellate cells by 99mTc-3PRGD2 scintigraphy targeting integrin αvβ3: a feasibility study

International Nuclear Information System (INIS)

Zhang, Xin; Xin, Jun; Shi, Yu; Xu, Weina; Yu, Shupeng; Yang, Zhiguang; Liu, Changping; Cao, Li; Guo, Qiyong

2015-01-01

Objective: Hepatic stellate cell (HSC) activation, which is accompanied by increased expression of integrin αvβ3, is an important factor in liver fibrogenesis. Molecular imaging targeting the integrin αvβ3 could provide a non-invasive method for evaluating the expression and the function of the integrin αvβ3 on the activated HSCs (aHSCs) in the injured liver, and then provide important prognostic information. 99m Tc-3PRGD2 is such a radiotracer specific for integrin αvβ3. In this study, we aimed to compare the differences in liver uptake and retention of the 99m Tc-3PRGD2 between normal liver and injured liver to evaluate the feasibility of 99m Tc-3PRGD2 scintigraphy for this purpose. Methods: We used planar scintigraphy to assess changes in integrin αvβ3 binding of intravenously-administered 99m Tc-3PRGD2 in the livers of rats with thioacetamide (TAA)-induced liver fibrosis compared with the controls. We co-injected cold c(RGDyK) with 99m Tc-3PRGD2 to assess the specific binding of the radiotracer. We performed Sirius red staining to assess liver fibrosis, immunofluorescent colocalization to identify the location of integrin αvβ3 expressed in the fibrotic liver, and we measured protein and messenger RNA expression of integrin αvβ3 and alpha smooth muscle actin (α-SMA) in the control and fibrotic livers. Results: The fibrotic livers showed enhanced 99m Tc-3PRGD2 uptake and retention. The radiotracer was demonstrated to bind specifically with the integrin αvβ3 mainly expressed on the aHSCs. The liver-to-heart ratio at 30 min post-injection was higher in the fibrotic livers than in the control livers (TAA, 1.98 ± 0.08 vs. control, 1.50 ± 0.12, p < 0.01). The liver t 1/2 was longer than in the controls (TAA, 27.07 ± 10.69 min vs. control, 12.67 ± 4.10 min, p < 0.01). The difference of heart t 1/2 between the two groups was not statistically significant (TAA, 3.13 ± 0.63 min vs. control, 3.41 ± 0.77 min, p = 0.94). Conclusions: 99m Tc-3PRGD2

Tailoring degradation of AZ31 alloy by surface pre-treatment and electrospun PCL fibrous coating

Energy Technology Data Exchange (ETDEWEB)

Hanas, T. [Medical Materials Laboratory, Indian Institute of Technology Madras, Chennai 600036 (India); School of Nano Science and Technology, National Institute of Technology Calicut, Calicut, Kerala 673601 (India); Sampath Kumar, T.S., E-mail: tssk@iitm.ac.in [Medical Materials Laboratory, Indian Institute of Technology Madras, Chennai 600036 (India); Perumal, Govindaraj; Doble, Mukesh [Department of Biotechnology - Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036 (India)

2016-08-01

AZ31 magnesium alloy was coated with polycaprolactone (PCL) nano-fibrous layer using electrospinning technique so as to control degradation in physiological environment. Before coating, the alloy was treated with HNO{sub 3} to have good adhesion between the coating and substrate. To elucidate the role of pre-treatment and coating, samples only with PCL coating as well as HNO{sub 3} treatment only were prepared for comparison. Best coating adhesion of 4B grade by ASTM D3359–09 tape test was observed for pre-treated samples. The effect of coating on in vitro degradation and biomineralization was studied using supersaturated simulated body fluid (SBF 5 ×). The weight loss and corrosion results obtained by immersion test showed that the combination of HNO{sub 3} pre-treatment and PCL coating is very effective in controlling the degradation rate and improving bioactivity. Cytotoxicity studies using L6 cells showed that PCL coated sample has better cell adhesion and proliferation compared to uncoated samples. Nano-fibrous PCL coating combined with prior acid treatment seems to be a promising method to tailor degradation rate with enhanced bioactivity of Mg alloys. - Highlights: • PCL electrospun coating on HNO{sub 3} pre-treated AZ31 alloy controls biodegradation. • Acid pre-treatment stabilizes the substrate - coating interface. • Electrospun porous coating improves biomineralization. • Coating similar to extracellular matrix enhances cell adhesion.

Coating of biodegradable magnesium alloy bone implants using nanostructured diopside (CaMgSi2O6)

Science.gov (United States)

Razavi, Mehdi; Fathi, Mohammadhossein; Savabi, Omid; Beni, Batoul Hashemi; Razavi, Seyed Mohammad; Vashaee, Daryoosh; Tayebi, Lobat

2014-01-01

Magnesium alloys with their biodegradable characteristic can be a very good candidate to be used in orthopedic implants. However, magnesium alloys may corrode and degrade too fast for applications in the bone healing procedure. In order to enhance the corrosion resistance and the in vitro bioactivity of a magnesium alloy, a nanostructured diopside (CaMgSi2O6) film was coated on AZ91 magnesium alloy through combined micro-arc oxidation (MAO) and electrophoretic deposition (EPD) methods. The crystalline structures, morphologies and compositions of the coated and uncoated substrates were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive spectroscopy. Polarization, electrochemical impedance spectroscopy, and immersion test in simulated body fluid (SBF) were employed to evaluate the corrosion resistance and the in vitro bioactivity of the samples. The results of our investigation showed that the nanostructured diopside coating deposited on the MAO layer increases the corrosion resistance and improves the in vitro bioactivity of the biodegradable magnesium alloy.

Coating of biodegradable magnesium alloy bone implants using nanostructured diopside (CaMgSi2O6)

International Nuclear Information System (INIS)

Razavi, Mehdi; Fathi, Mohammadhossein; Savabi, Omid; Beni, Batoul Hashemi; Razavi, Seyed Mohammad; Vashaee, Daryoosh

2014-01-01

Magnesium alloys with their biodegradable characteristic can be a very good candidate to be used in orthopedic implants. However, magnesium alloys may corrode and degrade too fast for applications in the bone healing procedure. In order to enhance the corrosion resistance and the in vitro bioactivity of a magnesium alloy, a nanostructured diopside (CaMgSi 2 O 6 ) film was coated on AZ91 magnesium alloy through combined micro-arc oxidation (MAO) and electrophoretic deposition (EPD) methods. The crystalline structures, morphologies and compositions of the coated and uncoated substrates were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive spectroscopy. Polarization, electrochemical impedance spectroscopy, and immersion test in simulated body fluid (SBF) were employed to evaluate the corrosion resistance and the in vitro bioactivity of the samples. The results of our investigation showed that the nanostructured diopside coating deposited on the MAO layer increases the corrosion resistance and improves the in vitro bioactivity of the biodegradable magnesium alloy.

Simple and high yielding method for preparing tissue specific extracellular matrix coatings for cell culture.

Science.gov (United States)

DeQuach, Jessica A; Mezzano, Valeria; Miglani, Amar; Lange, Stephan; Keller, Gordon M; Sheikh, Farah; Christman, Karen L

2010-09-27

The native extracellular matrix (ECM) consists of a highly complex, tissue-specific network of proteins and polysaccharides, which help regulate many cellular functions. Despite the complex nature of the ECM, in vitro cell-based studies traditionally assess cell behavior on single ECM component substrates, which do not adequately mimic the in vivo extracellular milieu. We present a simple approach for developing naturally derived ECM coatings for cell culture that provide important tissue-specific cues unlike traditional cell culture coatings, thereby enabling the maturation of committed C2C12 skeletal myoblast progenitors and human embryonic stem cells differentiated into cardiomyocytes. Here we show that natural muscle-specific coatings can (i) be derived from decellularized, solubilized adult porcine muscle, (ii) contain a complex mixture of ECM components including polysaccharides, (iii) adsorb onto tissue culture plastic and (iv) promote cell maturation of committed muscle progenitor and stem cells. This versatile method can create tissue-specific ECM coatings, which offer a promising platform for cell culture to more closely mimic the mature in vivo ECM microenvironment.

Phospho-Tyrosine(s) vs. Phosphatidylinositol Binding in Shc Mediated Integrin Signaling.

Science.gov (United States)

Lin, Xiaochen; Vinogradova, Olga

2015-04-01

The Shc adaptor protein, particularly its p52 isoform, has been identified as a primary signaling partner for the tyrosine(s)-phosphorylated cytoplasmic tails of activated β 3 integrins. Inspired by our recent structure of the Shc PTB domain in complex with a bi-phosphorylated peptide derived from β 3 cytoplasmic tail, we have initiated the investigation of Shc interaction with phospholipids of the membrane. We are particularly focused on PtdIns and their effects on Shc mediated integrin signaling in vitro . Here we present thermodynamic profiles and molecular details of the interactions between Shc, integrin, and PtdIns, all of which have been studied by ITC and solution NMR methods. A model of p52 Shc interaction with phosphorylated β 3 integrin cytoplasmic tail at the cytosolic face of the plasma membrane is proposed based on these data.

EMMPRIN regulates β1 integrin-mediated adhesion through Kindlin-3 in human melanoma cells.

Science.gov (United States)

Delyon, Julie; Khayati, Farah; Djaafri, Ibtissem; Podgorniak, Marie-Pierre; Sadoux, Aurélie; Setterblad, Niclas; Boutalbi, Zineb; Maouche, Kamel; Maskos, Uwe; Menashi, Suzanne; Lebbé, Céleste; Mourah, Samia

2015-06-01

EMMPRIN is known to promote tumor invasion through extracellular matrix (ECM) degradation. Here we report that EMMPRIN can regulate melanoma cell adhesion to the ECM through an interaction with β1 integrin involving kindlin-3. In this study, EMMPRIN knockdown in the human melanoma cell line M10 using siRNA decreased cell invasion and significantly increased cell adhesion and spreading. A morphological change from a round to a spread shape was observed associated with enhanced phalloidin-labelled actin staining. In situ proximity ligation assay and co-immunoprecipitation revealed that EMMPRIN silencing increased the interaction of β1 integrin with kindlin-3, a focal adhesion protein. This was associated with an increase in β1 integrin activation and a decrease in the phosphorylation of the downstream integrin kinase FAK. Moreover, the expression at both the transcript and protein level of kindlin-3 and of β1 integrin was inversely regulated by EMMPRIN. EMMPRIN did not regulate either talin expression or its interaction with β1 integrin. These results are consistent with our in vivo demonstration that EMMPRIN inhibition increased β1 integrin activation and its interaction with kindlin-3. To conclude, these findings reveal a new role of EMMPRIN in tumor cell migration through ß1 integrin/kindlin-3-mediated adhesion pathway. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Highly Specific Binding on Antifouling Zwitterionic Polymer-Coated Microbeads as Measured by Flow Cytometry.

Science.gov (United States)

van Andel, Esther; de Bus, Ian; Tijhaar, Edwin J; Smulders, Maarten M J; Savelkoul, Huub F J; Zuilhof, Han

2017-11-08

Micron- and nano-sized particles are extensively used in various biomedical applications. However, their performance is often drastically hampered by the nonspecific adsorption of biomolecules, a process called biofouling, which can cause false-positive and false-negative outcomes in diagnostic tests. Although antifouling coatings have been extensively studied on flat surfaces, their use on micro- and nanoparticles remains largely unexplored, despite the widespread experimental (specifically, clinical) uncertainties that arise because of biofouling. Here, we describe the preparation of magnetic micron-sized beads coated with zwitterionic sulfobetaine polymer brushes that display strong antifouling characteristics. These coated beads can then be equipped with recognition elements of choice, to enable the specific binding of target molecules. First, we present a proof of principle with biotin-functionalized beads that are able to specifically bind fluorescently labeled streptavidin from a complex mixture of serum proteins. Moreover, we show the versatility of the method by demonstrating that it is also possible to functionalize the beads with mannose moieties to specifically bind the carbohydrate-binding protein concanavalin A. Flow cytometry was used to show that thus-modified beads only bind specifically targeted proteins, with minimal/near-zero nonspecific protein adsorption from other proteins that are present. These antifouling zwitterionic polymer-coated beads, therefore, provide a significant advancement for the many bead-based diagnostic and other biosensing applications that require stringent antifouling conditions.

Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure

Science.gov (United States)

Maniotis, A. J.; Chen, C. S.; Ingber, D. E.

1997-01-01

We report here that living cells and nuclei are hard-wired such that a mechanical tug on cell surface receptors can immediately change the organization of molecular assemblies in the cytoplasm and nucleus. When integrins were pulled by micromanipulating bound microbeads or micropipettes, cytoskeletal filaments reoriented, nuclei distorted, and nucleoli redistributed along the axis of the applied tension field. These effects were specific for integrins, independent of cortical membrane distortion, and were mediated by direct linkages between the cytoskeleton and nucleus. Actin microfilaments mediated force transfer to the nucleus at low strain; however, tearing of the actin gel resulted with greater distortion. In contrast, intermediate filaments effectively mediated force transfer to the nucleus under both conditions. These filament systems also acted as molecular guy wires to mechanically stiffen the nucleus and anchor it in place, whereas microtubules acted to hold open the intermediate filament lattice and to stabilize the nucleus against lateral compression. Molecular connections between integrins, cytoskeletal filaments, and nuclear scaffolds may therefore provide a discrete path for mechanical signal transfer through cells as well as a mechanism for producing integrated changes in cell and nuclear structure in response to changes in extracellular matrix adhesivity or mechanics.

Antagonizing the alpha(4)beta(1) Integrin, but Not alpha(4)beta(7), Inhibits Leukocytic Infiltration of the Central Nervous System in Rhesus Monkey Experimental Autoimmune Encephalomyelitis

NARCIS (Netherlands)

Haanstra, Krista G.; Hofman, Sam O.; Estevao, Dave M. Lopes; Blezer, Erwin L. A.; Bauer, Jan; Yang, Li-Li; Wyant, Tim; Csizmadia, Vilmos; 't Hart, Bert A.; Fedyk, Eric R.

2013-01-01

The immune system is characterized by the preferential migration of lymphocytes through specific tissues (i.e., tissue tropism). Tissue tropism is mediated, in part, by the alpha(4) integrins expressed by T lymphocytes. The alpha(4)beta(1) integrin mediates migration of memory T lymphocytes into the

Phoyunnanin E inhibits migration of non-small cell lung cancer cells via suppression of epithelial-to-mesenchymal transition and integrin αv and integrin β3.

Science.gov (United States)

Petpiroon, Nareerat; Sritularak, Boonchoo; Chanvorachote, Pithi

2017-12-29

The conversion of the epithelial phenotype of cancer cells into cells with a mesenchymal phenotype-so-called epithelial-mesenchymal transition (EMT)-has been shown to enhance the capacity of the cells to disseminate throughout the body. EMT is therefore becoming a potential target for anti-cancer drug discovery. Here, we showed that phoyunnanin E, a compound isolated from Dendrobium venustum, possesses anti-migration activity and addressed its mechanism of action. The cytotoxic and proliferative effects of phoyunnanin E on human non-small cell lung cancer-derived H460, H292, and A549 cells and human keratinocyte HaCaT cells were investigated by MTT assay. The effect of phoyunnanin E on EMT was evaluated by determining the colony formation and EMT markers. The migration and invasion of H460, H292, A549 and HaCaT cells was evaluated by wound healing assay and transwell invasion assay, respectively. EMT markers, integrins and migration-associated proteins were examined by western blot analysis. Phoyunnanin E at the concentrations of 5 and 10 μM, which are non-toxic to H460, H292, A549 and HaCaT cells showed good potential to inhibit the migratory activity of three types of human lung cancer cells. The anti-migration effect of phoyunnanin E was shown to relate to the suppressed EMT phenotypes, including growth in anchorage-independent condition, cell motility, and EMT-specific protein markers (N-cadherin, vimentin, slug, and snail). In addition to EMT suppression, we found that phoyunnanin E treatment with 5 and 10 μM could decrease the cellular level of integrin αv and integrin β3, these integrins are frequently up-regulated in highly metastatic tumor cells. We further characterized the regulatory proteins in cell migration and found that the cells treated with phoyunnanin E exhibited a significantly lower level of phosphorylated focal adhesion kinase (p-FAK) and phosphorylated ATP-dependent tyrosine kinase (p-AKT), and their downstream effectors (including

Identification of a seed coat-specific promoter fragment from the Arabidopsis MUCILAGE-MODIFIED4 gene.

Science.gov (United States)

Dean, Gillian H; Jin, Zhaoqing; Shi, Lin; Esfandiari, Elahe; McGee, Robert; Nabata, Kylie; Lee, Tiffany; Kunst, Ljerka; Western, Tamara L; Haughn, George W

2017-09-01

The Arabidopsis seed coat-specific promoter fragment described is an important tool for basic and applied research in Brassicaceae species. During differentiation, the epidermal cells of the Arabidopsis seed coat produce and secrete large quantities of mucilage. On hydration of mature seeds, this mucilage becomes easily accessible as it is extruded to form a tightly attached halo at the seed surface. Mucilage is composed mainly of pectin, and also contains the key cell wall components cellulose, hemicellulose, and proteins, making it a valuable model for studying numerous aspects of cell wall biology. Seed coat-specific promoters are an important tool that can be used to assess the effects of expressing biosynthetic enzymes and diverse cell wall-modifying proteins on mucilage structure and function. Additionally, they can be used for production of easily accessible recombinant proteins of commercial interest. The MUCILAGE-MODIFIED4 (MUM4) gene is expressed in a wide variety of plant tissues and is strongly up-regulated in the seed coat during mucilage synthesis, implying the presence of a seed coat-specific region in its promoter. Promoter deletion analysis facilitated isolation of a 308 base pair sequence (MUM4 0.3Pro ) that directs reporter gene expression in the seed coat cells of both Arabidopsis and Camelina sativa, and is regulated by the same transcription factor cascade as endogenous MUM4. Therefore, MUM4 0.3Pro is a promoter fragment that serves as a new tool for seed coat biology research.

Vesicle-associated membrane protein 2 mediates trafficking of α5β1 integrin to the plasma membrane

International Nuclear Information System (INIS)

Hasan, Nazarul; Hu, Chuan

2010-01-01

Integrins are major receptors for cell adhesion to the extracellular matrix (ECM). As transmembrane proteins, the levels of integrins at the plasma membrane or the cell surface are ultimately determined by the balance between two vesicle trafficking events: endocytosis of integrins at the plasma membrane and exocytosis of the vesicles that transport integrins. Here, we report that vesicle-associated membrane protein 2 (VAMP2), a SNARE protein that mediates vesicle fusion with the plasma membrane, is involved in the trafficking of α5β1 integrin. VAMP2 was present on vesicles containing endocytosed β1 integrin. Small interfering RNA (siRNA) silencing of VAMP2 markedly reduced cell surface α5β1 and inhibited cell adhesion and chemotactic migration to fibronectin, the ECM ligand of α5β1, without altering cell surface expression of α2β1 integrin or α3β1 integrin. By contrast, silencing of VAMP8, another SNARE protein, had no effect on cell surface expression of the integrins or cell adhesion to fibronectin. In addition, VAMP2-mediated trafficking is involved in cell adhesion to collagen but not to laminin. Consistent with disruption of integrin functions in cell proliferation and survival, VAMP2 silencing diminished proliferation and triggered apoptosis. Collectively, these data indicate that VAMP2 mediates the trafficking of α5β1 integrin to the plasma membrane and VAMP2-dependent integrin trafficking is critical in cell adhesion, migration and survival.

Synthesis and Characterization of Nano-Hydroxyapatite/mPEG-b-PCL Composite Coating on Nitinol Alloy

OpenAIRE

Mohamadreza Etminanfar; Jafar Khalil-Allafi; Kiyumars Jalili

2017-01-01

In this study the bioactivity of hydroxyapatite/poly(ε-caprolactone)–poly(ethylene glycol) bilayer coatings on Nitinol superelastic alloy was investigated. The surface of Nitinol alloy was activated by a thermo-chemical treatment and hydroxyapatite coating was electrodeposited on the alloy, followed by applying the polymer coating. The surface morphology of coatings was studied using FE-SEM and SEM. The data revealed that the hydroxyapatite coating is composed of one-dimensional nano sized fl...

The possibilities of atmospheric plasma-spraying application to obtain hydroxyapatite coatings on the stainless steel samples

Directory of Open Access Journals (Sweden)

Mihailović Marija D.

2013-01-01

Full Text Available For decades, the standard metallic materials for hip implants, besides the 316LVM stainless steel, were titanium- and cobalt/chromium-based alloys. Although bioinert, due to their corrosion resistance, they are not biocompatible. Contemporary surgical implants are not made just of bioinert metal anymore, but with deposited bioactive hydroxyapatite (HAp coating. Hydroxyapatite is chemically identical with the mineral constituent of bones and teeth, what besides its biocompatibility provides bioactivity as well. The HAp limitations are, however, weak tensile strength and low fatigue resistance for long term loadings, if used alone. This is the reason for HAp to be deposited onto the surgical implant, and to enable its bioactivity, what means intergrowth with bones, and therefore the long-lasting and mechanical stable non-cemented prosthesis. This is important predominantly because the need for such prostheses for younger population, and a better life quality. There are several contemporary techniques that have been used for deposition of these coatings onto the metal implant. The possibilities of atmospheric plasma-spraying for obtaining the stable HAp coatings on the 316LVM stainless steel, ordinary used as a standard material for hip implants production are presented in this paper. The coatings of a commercially available hydroxyapatite powder were plasma-sprayed onto the specimens of medical grade 316LVM stainless steel under various operating conditions. The optical microscopy was used for microstructure and porosity characterization, while coating morphology and Ca/P ratio were analyzed using SEM equipped with EDX. Coating microstructure varied from a porous to a glassy structure, depending on operating conditions applied and coating thickness. Coating porosity was determined to be at the lower required limit requested for the bone-coating intergrowth possibility, but nevertheless adhesion measurements showed good results. The Ca/P ratio was

Apolar Bioactive Fraction of Melipona scutellaris Geopropolis on Streptococcus mutans Biofilm.

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da Cunha, Marcos Guilherme; Franchin, Marcelo; Galvão, Lívia Câmara de Carvalho; Bueno-Silva, Bruno; Ikegaki, Masaharu; de Alencar, Severino Matias; Rosalen, Pedro Luiz

2013-01-01

The aim of this study was to evaluate the influence of the bioactive nonpolar fraction of geopropolis on Streptococcus mutans biofilm. The ethanolic extract of Melipona scutellaris geopropolis was subjected to a liquid-liquid partition, thus obtaining the bioactive hexane fraction (HF) possessing antimicrobial activity. The effects of HF on S. mutans UA159 biofilms generated on saliva-coated hydroxyapatite discs were analyzed by inhibition of formation, killing assay, and glycolytic pH-drop assays. Furthermore, biofilms treated with vehicle control and HF were analyzed by scanning electron microscopy (SEM). HF at 250  μ g/mL and 400  μ g/mL caused 38% and 53% reduction in the biomass of biofilm, respectively, when compared to vehicle control (P 0.05). In conclusion, the bioactive HF of geopropolis was promising to control the S. mutans biofilm formation, without affecting the microbial population but interfering with its structure by reducing the biochemical content of biofilm matrix.

Apolar Bioactive Fraction of Melipona scutellaris Geopropolis on Streptococcus mutans Biofilm

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Marcos Guilherme da Cunha

2013-01-01

Full Text Available The aim of this study was to evaluate the influence of the bioactive nonpolar fraction of geopropolis on Streptococcus mutans biofilm. The ethanolic extract of Melipona scutellaris geopropolis was subjected to a liquid-liquid partition, thus obtaining the bioactive hexane fraction (HF possessing antimicrobial activity. The effects of HF on S. mutans UA159 biofilms generated on saliva-coated hydroxyapatite discs were analyzed by inhibition of formation, killing assay, and glycolytic pH-drop assays. Furthermore, biofilms treated with vehicle control and HF were analyzed by scanning electron microscopy (SEM. HF at 250 μg/mL and 400 μg/mL caused 38% and 53% reduction in the biomass of biofilm, respectively, when compared to vehicle control (P0.05. In conclusion, the bioactive HF of geopropolis was promising to control the S. mutans biofilm formation, without affecting the microbial population but interfering with its structure by reducing the biochemical content of biofilm matrix.

An overview of the effects of thermal processing on bioactive glasses

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Bellucci D.

2010-01-01

Full Text Available Bioglass® 45S5 is widely used in biomedical applications due to its ability to bond to bone and even to soft tissues. The sintering ability of Bioglass® powders is a key factor from a technological point of view, since its govern the production of advanced devices, ranging from highly porous scaffolds to functionalized coatings. Unfortunately this particular glass composition is prone to crystallize at the temperature required for sintering and this may impair the bioactivity of the original glass. For these reasons, a prerequisite to tailor the fabrication of Bioglass®-derived implants is to understand the interaction between sintering, crystallization and bioactivity. In this work the structural transformations which occur during the heat treatment of Bioglass® are reviewed and a special attention is paid to the sintering and crystallization processes. Moreover the bioactivity of the final glass-ceramics is discussed and some alternative glass formulations are reported.

α6 Integrin and CD44 enrich for a primary keratinocyte population that displays resistance to UV-induced apoptosis.

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Helen Wray

Full Text Available Epidermal human keratinocytes are exposed to a wide range of environmental genotoxic insults, including the UV component of solar radiation. Epidermal homeostasis in response to cellular or tissue damage is maintained by a population of keratinocyte stem cells (KSC that reside in the basal layer of the epithelium. Using cell sorting based on cell-surface markers, we have identified a novel α6 integrin(high+/CD44(+ sub-population of basal keratinocytes. These α6 integrin(high+/CD44(+ keratinocytes have both high proliferative potential, form colonies in culture that have characteristics of holoclones and have a unique pattern of resistance to apoptosis induced by UVB radiation or by agents that induce single- or double strand DNA breaks. Resistance to UVB induced apoptosis in the α6 integrin(high+/CD44(+ cells involved increased expression of TAp63 and was overcome by PI-3 kinase inhibition. In marked contrast, the α6 integrin(high+/CD44(+ cells were sensitive to apoptosis induced by the cross-linking agent cisplatin, and imatinib inhibition of c-Abl blocked the ability of cisplatin to kill α6 integrin(high+/CD44(+ cells. Our findings reveal a population of basal keratinocytes with long-term proliferative properties that display specific patterns of apoptotic resistance that is dependent upon the genotoxic stimulus, and provide insights into how these cells can be targeted with chemotherapeutic agents.

Bidirectional remodeling of β1-integrin adhesions during chemotropic regulation of nerve growth

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Carlstrom Lucas P

2011-11-01

Full Text Available Abstract Background Chemotropic factors in the extracellular microenvironment guide nerve growth by acting on the growth cone located at the tip of extending axons. Growth cone extension requires the coordination of cytoskeleton-dependent membrane protrusion and dynamic adhesion to the extracellular matrix, yet how chemotropic factors regulate these events remains an outstanding question. We demonstrated previously that the inhibitory factor myelin-associated glycoprotein (MAG triggers endocytic removal of the adhesion receptor β1-integrin from the growth cone surface membrane to negatively remodel substrate adhesions during chemorepulsion. Here, we tested how a neurotrophin might affect integrin adhesions. Results We report that brain-derived neurotropic factor (BDNF positively regulates the formation of substrate adhesions in axonal growth cones during stimulated outgrowth and prevents removal of β1-integrin adhesions by MAG. Treatment of Xenopus spinal neurons with BDNF rapidly triggered β1-integrin clustering and induced the dynamic formation of nascent vinculin-containing adhesion complexes in the growth cone periphery. Both the formation of nascent β1-integrin adhesions and the stimulation of axon extension by BDNF required cytoplasmic calcium ion signaling and integrin activation at the cell surface. Exposure to MAG decreased the number of β1-integrin adhesions in the growth cone during inhibition of axon extension. In contrast, the BDNF-induced adhesions were resistant to negative remodeling by MAG, correlating with the ability of BDNF pretreatment to counteract MAG-inhibition of axon extension. Pre-exposure to MAG prevented the BDNF-induced formation of β1-integrin adhesions and blocked the stimulation of axon extension by BDNF. Conclusions Altogether, these findings demonstrate the neurotrophin-dependent formation of integrin-based adhesions in the growth cone and reveal how a positive regulator of substrate adhesions can block

Syndecan-4 Phosphorylation Is a Control Point for Integrin Recycling

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Morgan, Mark R.; Hamidi, Hellyeh; Bass, Mark D.; Warwood, Stacey; Ballestrem, Christoph; Humphries, Martin J.

2013-01-01

Summary Precise spatiotemporal coordination of integrin adhesion complex dynamics is essential for efficient cell migration. For cells adherent to fibronectin, differential engagement of α5β1 and αVβ3 integrins is used to elicit changes in adhesion complex stability, mechanosensation, matrix assembly, and migration, but the mechanisms responsible for receptor regulation have remained largely obscure. We identify phosphorylation of the membrane-intercalated proteoglycan syndecan-4 as an essential switch controlling integrin recycling. Src phosphorylates syndecan-4 and, by driving syntenin binding, leads to suppression of Arf6 activity and recycling of αVβ3 to the plasma membrane at the expense of α5β1. The resultant elevation in αVβ3 engagement promotes stabilization of focal adhesions. Conversely, abrogation of syndecan-4 phosphorylation drives surface expression of α5β1, destabilizes adhesion complexes, and disrupts cell migration. These data identify the dynamic spatiotemporal regulation of Src-mediated syndecan-4 phosphorylation as an essential switch controlling integrin trafficking and adhesion dynamics to promote efficient cell migration. PMID:23453597

Integrin-targeting thermally cross-linked superparamagnetic iron oxide nanoparticles for combined cancer imaging and drug delivery

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Yu, Mi Kyung; Park, Jinho; Jon, Sangyong [School of Life Sciences, Gwangju Institute of Science and Technology, 261 Chemdangwagi-ro, Gwangju 500-712 (Korea, Republic of); Jeong, Yong Yeon [Department of Diagnostic Radiology, Jeonnam National University Hwasun Hospital, 160 Ilsim-ri, Hwasun-eup, Jeonnam 519-809 (Korea, Republic of); Moon, Woo Kyung, E-mail: syjon@gist.ac.kr [Diagnostic Radiology, Seoul National University Hospital and the Institute of Radiation Medicine, Medical Research Center Seoul National University, Seoul 110-744 (Korea, Republic of)

2010-10-15

We report multifunctional nanoparticles that are capable of cancer targeting and simultaneous cancer imaging and therapy. The nanoparticles are composed of cyclic arginine-glycine-aspartic acid (cRGD) peptide ligand bioconjugated thermally cross-linked superparamagnetic iron oxide nanoparticles (TCL-SPION) that enable loading of the anticancer drug doxorubicin (Dox). The cyclic RGD-conjugated TCL-SPION (cRGD{sub T}CL-SPION) had a mean hydrodynamic size of 34 {+-} 8 nm with approximately 0.39 wt% of cyclic RGD attached to the surface of the nanoparticles. The cRGD{sub T}CL-SPION exhibited preferential binding towards target cancer cells (U87MG, integrin {alpha}{sub v{beta}3} +) when analyzed by T{sub 2}-weighted magnetic resonance (MR) imaging. When Dox was loaded onto the polymeric coating layers of cRGD{sub T}CL-SPION via ionic interaction, the resulting Dox-loaded cRGD{sub T}CL-SPION (Dox-cRGD{sub T}CL-SPION) showed much higher cytotoxicity in U87MG cells than Dox-TCL-SPION lacking cRGD (IC{sub 50} value of 0.02 {mu}M versus 0.12 {mu}M). These results suggest that Dox-cRGD{sub T}CL-SPION has potential for use as an integrin-targeted, combined imaging and therapeutic agent.

ADAM2 interactions with mouse eggs and cell lines expressing α4/α9 (ITGA4/ITGA9 integrins: implications for integrin-based adhesion and fertilization.

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Ulyana V Desiderio

2010-10-01

Full Text Available Integrins are heterodimeric cell adhesion molecules, with 18 α (ITGA and eight β (ITGB subunits forming 24 heterodimers classified into five families. Certain integrins, especially the α(4/α(9 (ITGA4/ITGA9 family, interact with members of the ADAM (a disintegrin and metalloprotease family. ADAM2 is among the better characterized and also of interest because of its role in sperm function. Having shown that ITGA9 on mouse eggs participates in mouse sperm-egg interactions, we sought to characterize ITGA4/ITGA9-ADAM2 interactions.An anti-β(1/ITGB1 function-blocking antibody that reduces sperm-egg binding significantly inhibited ADAM2 binding to mouse eggs. Analysis of integrin subunit expression indicates that mouse eggs could express at least ten different integrins, five in the RGD-binding family, two in the laminin-binding family, two in the collagen-binding family, and ITGA9-ITGB1. Adhesion assays to characterize ADAM2 interactions with ITGA4/ITGA9 family members produced the surprising result that RPMI 8866 cell adhesion to ADAM2 was inhibited by an anti-ITGA9 antibody, noteworthy because ITGA9 has only been reported to dimerize with ITGB1, and RPMI 8866 cells lack detectable ITGB1. Antibody and siRNA studies demonstrate that ITGB7 is the β subunit contributing to RPMI 8866 adhesion to ADAM2.These data indicate that a novel integrin α-β combination, ITGA9-ITGB7 (α(9β(7, in RPMI 8866 cells functions as a binding partner for ADAM2. ITGA9 had previously only been reported to dimerize with ITGB1. Although ITGA9-ITGB7 is unlikely to be a widely expressed integrin and appears to be the result of "compensatory dimerization" occurring in the context of little/no ITGB1 expression, the data indicate that ITGA9-ITGB7 functions as an ADAM binding partner in certain cellular contexts, with implications for mammalian fertilization and integrin function.

Role of β1 Integrin in Tissue Homing of Neutrophils During Sepsis

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Sarangi, Pranita P.; Hyun, Young-Min; Lerman, Yelena V.; Pietropaoli, Anthony P.; Kim, Minsoo

2012-01-01

Aberrant activation of neutrophils during sepsis results in the widespread release of pro-inflammatory mediators, leading to multi-organ system failure and death. However, aberrant activation of neutrophils during sepsis results in the widespread release of harmful inflammatory mediators causing host tissue injuries that can lead to multi organ system failure and death. One of the pivotal components of neutrophil migration during inflammation is the expression of surface integrins. In this study, we show that administration of a cyclic analog of RGD peptide (Arg-Gly-Asp) significantly reduced the number of tissue-invading neutrophils and the degree of sepsis-induced lethality in mice as compared to control peptide. Secondly, β1 integrin (CD29) was highly up-regulated on the neutrophils isolated from both septic patients and animals. Finally, conditional genetic ablation of β1 integrin from granulocytes also improved survival and bacterial clearance in septic animals Thus, our results indicate that expression of β1 integrin is important for modulating neutrophil trafficking during sepsis, and that therapeutics designed against β1 integrins may be beneficial. PMID:22683734

Interactions of foot-and-mouth disease virus with soluble bovine alphaVbeta3 and alphaVbeta6 integrins.

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Duque, Hernando; LaRocco, Michael; Golde, William T; Baxt, Barry

2004-09-01

At least four members of the integrin family of receptors, alphaVbeta1, alphaVbeta3, alphaVbeta6, and alphaVbeta8, have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro. Our investigators have recently shown that the efficiency of receptor usage appears to be related to the viral serotype and may be influenced by structural differences on the viral surface (H. Duque and B. Baxt, J. Virol. 77:2500-2511, 2003). To further examine these differences, we generated soluble alphaVbeta3 and alphaVbeta6 integrins. cDNA plasmids encoding the individual complete integrin alphaV, beta3, and beta6 subunits were used to amplify sequences encoding the subunits' signal peptide and ectodomain, resulting in subunits lacking transmembrane and cytoplasmic domains. COS-1 cells were transfected with plasmids encoding the soluble alphaV subunit and either the soluble beta3 or beta6 subunit and labeled with [35S]methionine-cysteine. Complete subunit heterodimeric integrins were secreted into the medium, as determined by radioimmunoprecipitation with specific monoclonal and polyclonal antibodies. For the examination of the integrins' biological activities, stable cell lines producing the soluble integrins were generated in HEK 293A cells. In the presence of divalent cations, soluble alphaVbeta6 bound to representatives of type A or O viruses, immobilized on plastic dishes, and significantly inhibited viral replication, as determined by plaque reduction assays. In contrast, soluble alphaVbeta3 was unable to bind to immobilized virus of either serotype; however, virus bound to the immobilized integrin, suggesting that FMDV binding to alphaVbeta3 is a low-affinity interaction. In addition, soluble alphaVbeta3 did not neutralize virus infectivity. Incubation of soluble alphaVbeta6 with labeled type A12 or O1 resulted in a significant inhibition of virus adsorption to BHK cells, while soluble alphaVbeta3 caused a low (20 to 30%), but consistent, inhibition of virus

α6β1- and αV-integrins are required for long-term self-renewal of murine embryonic stem cells in the absence of LIF.

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Cattavarayane, Sandhanakrishnan; Palovuori, Riitta; Tanjore Ramanathan, Jayendrakishore; Manninen, Aki

2015-02-27

The growth properties and self-renewal capacity of embryonic stem (ES) cells are regulated by their immediate microenvironment such as the extracellular matrix (ECM). Integrins, a central family of cellular ECM receptors, have been implicated in these processes but their specific role in ES cell self-renewal remains unclear. Here we have studied the effects of different ECM substrates and integrins in mouse ES cells in the absence of Leukemia Inhibitory Factor (LIF) using short-term assays as well as long-term cultures. Removal of LIF from ES cell culture medium induced morphological differentiation of ES cells into polarized epistem cell-like cells. These cells maintained epithelial morphology and expression of key stemness markers for at least 10 passages in the absence of LIF when cultured on laminin, fibronectin or collagen IV substrates. The specific functional roles of α6-, αV- and β1-integrin subunits were dissected using stable lentivirus-mediated RNAi methodology. β1-integrins were required for ES cell survival in long-term cultures and for the maintenance of stem cell marker expression. Inhibition of α6-integrin expression compromised self-renewal on collagen while αV-integrins were required for robust ES cell adhesion on laminin. Analysis of the stemness marker expression revealed subtle differences between α6- and αV-depleted ES cells but the expression of both was required for optimal self-renewal in long-term ES cell cultures. In the absence of LIF, long-term ES cell cultures adapt an epistem cell-like epithelial phenotype and retain the expression of multiple stem cell markers. Long-term maintenance of such self-renewing cultures depends on the expression of β1-, α6- and αV-integrins.

Lumican alleviates hypertrophic scarring by suppressing integrin-FAK signaling

International Nuclear Information System (INIS)

Zhao, Yuqian; Li, Xueyong; Xu, Xiaoli; He, Zhi; Cui, Lei; Lv, Xiaoxing