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1.
Foods ; 11(7)2022 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-35407124

RESUMO

Bacillus cereus is relatively resistant to pasteurization. We assessed the risk of B. cereus growth during warming and subsequent storage of pasteurized banked milk (PBM) in the warmed state using a predictive mathematical model. Holder pasteurization followed by storage below -18 °C was used. Temperature maps, water activity values, and B. cereus growth in artificially inoculated PBM were obtained during a simulation of manipulation of PBM after its release from a Human Milk Bank. As a real risk level, we chose a B. cereus concentration of 100 CFU/mL; the risk was assessed for three cases: 1. For an immediate post-pasteurization B. cereus concentration below 1 CFU/mL (level of detection); 2. For a B. cereus concentration of 10 CFU/mL, which is allowed in some countries; 3. For a B. cereus concentration of 50 CFU/mL, which is approved for milk formulas. In the first and second cases, no risk was detected after 1 h of storage in the warmed state, while after 2 h of storage, B. cereus concentrations of 102 CFU/mL were occasionally encountered. In the third case, exceeding the B. cereus concentration of 102 CFU/mL could be regularly expected after 2 h of storage. Based on these results, we recommend that post-pasteurization bacteriological analysis be performed as recommended by the European Milk Bank Association (EMBA) and using warmed PBM within 1 h after warming (no exceptions).

2.
Polymers (Basel) ; 14(3)2022 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-35160567

RESUMO

This article describes 1D extension tests on bovine collagen samples (8% collagen in water). At such a high collagen concentration, the mechanical properties of semi-solid samples can be approximated by hyperelastic models (two-parametric HGO and Misof models were used), or simply by Hooke's law and the modulus of elasticity E. The experiments confirm a significant increase in the E-modulus of the samples irradiated with high-energy electrons. The modulus E ~ 9 kPa of non-irradiated samples increases monotonically up to E ~ 250 kPa for samples absorbing an e-beam dose of ~3300 Gy. This amplification is attributed to the formation of cross-links by irradiation. However, E-modulus can be increased not only by irradiation but also by exposure to a high strain rate. For example, soft isotropic collagen extruded through a 200 mm long capillary increases the modulus of elasticity from 9 kPa to 30 kPa, and the increase is almost isotropic. This stiffening occurs when the corrugated collagen fibers are straightened and are aligned in the flow direction. It seems that the permanent structural changes caused by extrusion mitigate the effects of the ex post applied irradiation. Irradiation of extruded samples by 3300 Gy increases the modulus of E-elasticity only three times (from 30 kPa to approximately 90 kPa). Extruded and ex post irradiated samples show slight anisotropy (the stiffness in the longitudinal direction is on an average greater than the transverse stiffness).

3.
Foods ; 11(4)2022 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-35206086

RESUMO

This Special Issue is focused on the use of modern food processing technologies to retain the highest possible content of health-promoting compounds in raw foods [...].

4.
Foods ; 11(2)2022 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-35053954

RESUMO

High pressure processing (HPP) is a cold pasteurization technology by which products, prepacked in their final package, are introduced to a vessel and subjected to a high level of isostatic pressure (300-600 MPa). High-pressure treatment of fruit, vegetable and fresh herb homogenate products offers us nearly fresh products in regard to sensorial and nutritional quality of original raw materials, representing relatively stable and safe source of nutrients, vitamins, minerals and health effective components. Such components can play an important role as a preventive tool against the start of illnesses, namely in the elderly. An overview of several food HPP products, namely of fruit and vegetable origin, marketed successfully around the world is presented. Effects of HPP and HPP plus heat on key spoilage and pathogenic microorganisms, including the resistant spore form and fruit/vegetable endogenous enzymes are reviewed, including the effect on the product quality. Part of the paper is devoted to the industrial equipment available for factories manufacturing HPP treated products.

5.
Foods ; 10(12)2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34945507

RESUMO

A systematic study, performed from 2017-2020 looked at the rate of positive post-pasteurization B. cereus findings, the quantity of B. cereus in pasteurized banked human milk (PBM), and the rate of B. cereus toxicogenic isolates from PBM. During the study period, 6815.71 L (30,943 tested bottles) of PBM were tested, with an average amount per year of 1703.93 L (7736 tested bottles). The PBM discard rate per year due to bacterial contamination varied between 8.7-10.0% and contamination with B. cereus was the most frequent reason. The total number of B. cereus positive tests was 2739 and the proportion of its positivity from all positive tests was between 56.7-66.6%. The prevalence of B. cereus positive tests rose significantly in the summer months. The production of enterotoxin was found in 3 of the 20 tested samples (15.0%). The B. cereus CFU-quantities in the PBM were below 10 CFU/mL in 80% of cases (16 of 20 samples tested). The quantitative data can be used in the risk assessment of cold storage of PBM at temperatures above zero and manipulation of PBM prior to its administration.

6.
ACS Appl Mater Interfaces ; 13(50): 60612-60624, 2021 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-34902239

RESUMO

New analytical techniques that overcome major drawbacks of current routinely used viral infection diagnosis methods, i.e., the long analysis time and laboriousness of real-time reverse-transcription polymerase chain reaction (qRT-PCR) and the insufficient sensitivity of "antigen tests", are urgently needed in the context of SARS-CoV-2 and other highly contagious viruses. Here, we report on an antifouling terpolymer-brush biointerface that enables the rapid and sensitive detection of SARS-CoV-2 in untreated clinical samples. The developed biointerface carries a tailored composition of zwitterionic and non-ionic moieties and allows for the significant improvement of antifouling capabilities when postmodified with biorecognition elements and exposed to complex media. When deployed on a surface of piezoelectric sensor and postmodified with human-cell-expressed antibodies specific to the nucleocapsid (N) protein of SARS-CoV-2, it made possible the quantitative analysis of untreated samples by a direct detection assay format without the need of additional amplification steps. Natively occurring N-protein-vRNA complexes, usually disrupted during the sample pre-treatment steps, were detected in the untreated clinical samples. This biosensor design improved the bioassay sensitivity to a clinically relevant limit of detection of 1.3 × 104 PFU/mL within a detection time of only 20 min. The high specificity toward N-protein-vRNA complexes was validated both by mass spectrometry and qRT-PCR. The performance characteristics were confirmed by qRT-PCR through a comparative study using a set of clinical nasopharyngeal swab samples. We further demonstrate the extraordinary fouling resistance of this biointerface through exposure to other commonly used crude biological samples (including blood plasma, oropharyngeal, stool, and nasopharyngeal swabs), measured via both the surface plasmon resonance and piezoelectric measurements, which highlights the potential to serve as a generic platform for a wide range of biosensing applications.


Assuntos
Teste para COVID-19 , COVID-19/diagnóstico , Proteínas do Nucleocapsídeo de Coronavírus/química , Mucosa Nasal/virologia , Polímeros/química , RNA Viral/metabolismo , SARS-CoV-2 , Incrustação Biológica , Bioensaio , Técnicas Biossensoriais , Humanos , Íons , Limite de Detecção , Espectrometria de Massas , Nasofaringe/virologia , Fosfoproteínas/química , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Manejo de Espécimes
7.
Foods ; 10(8)2021 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-34441704

RESUMO

Broccoli sprouts contain 10-100 times higher levels of sulforaphane than mature plants, something that has been well known since 1997. Sulforaphane has a whole range of unique biological properties, and it is especially an inducer of phase 2 detoxication enzymes. Therefore, its use has been intensively studied in the field of health and nutrition. The formation of sulforaphane is controlled by the epithiospecifier protein, a myrosinase co-factor, which is temperature-specific. This paper studies the influence of temperature, heating time, the addition of myrosinase in the form of Raphanus sativus sprouts in constant ratio to broccoli sprouts, and other technological steps on the final sulforaphane content in broccoli sprout homogenates. These technological steps are very important for preserving sulforaphane in broccoli sprouts, but there are some limitations concerning the amount of sulforaphane. We focused, therefore, on the extraction process, using suitable ß-cyclodextrin, hexane and ethanol, with the goal of increasing the amount of sulforaphane in the final extract, thus stabilizing it and reducing the required amount sulforaphane needed, e.g., as a dietary supplement.

8.
Foods ; 10(8)2021 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-34441719

RESUMO

The aim of this work was to compare selected physicochemical properties of air dried 'Golden Delicious' apples, pretreated either by high-pressure processing (HPP), ultrasound (US) or pulsed electric field (PEF). Following parameters of pretreatment were used: HPP-400 MPa for 15 min, US-21 kHz, 180 W for 45 min, PEF-1 kV/cm, 3.5 kJ/kg. The quality of materials was evaluated by their rehydration properties, hygroscopicity, color and total phenolic content. To compare the effectiveness of the utilized methods, determined properties were expressed as relative comparison values against the reference sample obtained without any pretreatment in the same conditions. The performed research demonstrated that properties can be shaped by the application of proper pretreatment methods. For instance, PEF was shown to be the best method for improving water uptake during rehydration, whereas HPP was the most effective in decreasing hygroscopic properties in comparison with untreated dried apples. Among the investigated methods, HPP resulted in the deepest browning and thus total color difference, while the effects of US and PEF were comparable. For all pretreated dried apples, the total phenolic content was lower when compared with reference material, though the smallest drop was found in sonicated samples.

9.
Antibiotics (Basel) ; 10(6)2021 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-34204644

RESUMO

Staphylococcus (S.) aureus is an important causative agent of wound infections with increasing incidence in the past decades. Specifically, the emergence of methicillin-resistant S. aureus (MRSA) causes serious problems, especially in nosocomial infections. Therefore, there is an urgent need to develop of alternative or supportive antimicrobial therapeutic modalities to meet these challenges. Purified compounds from hops have previously shown promising antimicrobial effects against MRSA isolates in vitro. In this study, purified beta-acids from hops were tested for their potential antimicrobial and healing properties using a porcine model of wounds infected by MRSA. The results show highly significant antimicrobial effects of the active substance in both the powder and Ambiderman-based application forms compared to both no-treatment control and treatment with Framycoin. Moreover, the macroscopic evaluation of the wounds during the treatment using the standardized Wound Healing Continuum indicated positive effects of the beta-acids on the overall wound healing. This is further supported by the microscopic data, which showed a clear improvement of the inflammatory parameters in the wounds treated by beta-acids. Thus, using the porcine model, we demonstrate significant therapeutic effects of hops compounds in the management of wounds infected by MRSA. Beta-acids from hops, therefore, represent a suitable candidate for the treatment of non-responsive nosocomial tissue infections by MRSA.

10.
Antibiotics (Basel) ; 10(4)2021 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-33917416

RESUMO

Clostridioides (C.) difficile is an important causative pathogen of nosocomial gastrointestinal infections in humans with an increasing incidence, morbidity, and mortality. The available treatment options against this pathogen are limited. The standard antibiotics are expensive, can promote emerging resistance, and the recurrence rate of the infection is high. Therefore, there is an urgent need for new approaches to meet these challenges. One of the possible treatment alternatives is to use compounds available in commonly used plants. In this study, purified extracts isolated from hops-alpha and beta acids and xanthohumol-were tested in vivo for their inhibitory effect against C. difficile. A rat model of the peroral intestinal infection by C. difficile has been developed. The results show that both xanthohumol and beta acids from hops exert a notable antimicrobial effect in the C. difficile infection. The xanthohumol application showed the most pronounced antimicrobial effect together with an improvement of local inflammatory signs in the large intestine. Thus, the hops compounds represent promising antimicrobial agents for the treatment of intestinal infections caused by C. difficile.

11.
J Food Sci Technol ; 58(2): 581-585, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33568851

RESUMO

Lignans are members of a broad group of plant phenols that can positively affect human health. They occur in negligible quantities in processed foodstuffs such as lager beer. The aim of this work was to utilize the high levels of lignans in the knots of spruce trees (Picea abies) to increase the lignans content in beer, without negatively impacting the natural taste and aroma. By means of lignans addition in the forms of spruce knot chips or different extracts made from spruce knots during the wort boiling were produced beer and beer-based beverages with lignans content ranging from 34 to 174 mg/L.

12.
Biomolecules ; 10(8)2020 08 05.
Artigo em Inglês | MEDLINE | ID: mdl-32764330

RESUMO

Cells are continuously sensing their microenvironment and subsequently respond to different physicochemical cues by the activation or inhibition of different signaling pathways. To study a very complex cellular response, it is necessary to diminish background environmental influences and highlight the particular event. However, surface-driven nonspecific interactions of the abundant biomolecules from the environment influence the targeted cell response significantly. Yes-associated protein (YAP) translocation may serve as a marker of human hepatocellular carcinoma (Huh7) cell responses to the extracellular matrix and surface-mediated stresses. Here, we propose a platform of tunable functionable antifouling poly(carboxybetain) (pCB)-based brushes to achieve a molecularly clean background for studying arginine, glycine, and aspartic acid (RGD)-induced YAP-connected mechanotransduction. Using two different sets of RGD-functionalized zwitterionic antifouling coatings with varying compositions of the antifouling layer, a clear correlation of YAP distribution with RGD functionalization concentrations was observed. On the other hand, commonly used surface passivation by the oligo(ethylene glycol)-based self-assembled monolayer (SAM) shows no potential to induce dependency of the YAP distribution on RGD concentrations. The results indicate that the antifouling background is a crucial component of surface-based cellular response studies, and pCB-based zwitterionic antifouling brush architectures may serve as a potential next-generation easily functionable surface platform for the monitoring and quantification of cellular processes.


Assuntos
Incrustação Biológica/prevenção & controle , Materiais Revestidos Biocompatíveis/química , Mecanotransdução Celular , Acrilamidas/química , Linhagem Celular Tumoral , Matriz Extracelular/metabolismo , Humanos , Oligopeptídeos/química , Proteínas Proto-Oncogênicas c-yes/metabolismo , Estresse Mecânico
13.
Langmuir ; 36(29): 8485-8493, 2020 07 28.
Artigo em Inglês | MEDLINE | ID: mdl-32506911

RESUMO

Polymer brushes not only represent emerging surface platforms for numerous bioanalytical and biological applications but also create advanced surface-tethered systems to mimic real-life biological processes. In particular, zwitterionic and nonionic polymer brushes have been intensively studied because of their extraordinary resistance to nonspecific adsorption of biomolecules (antifouling characteristics) as well as the ability to be functionalized with bioactive molecules. However, the relation between antifouling behavior in real-world biological media and structural changes of polymer brushes induced by surface preconditioning in different environments remains unexplored. In this work, we use multiple methods to study the structural properties of numerous brushes under variable ionic concentrations and determine the impact of these changes on resistance to fouling from undiluted blood plasma. We describe different mechanisms of swelling, depending on both the polymer brush coating properties and the environmental conditions that affect changes in both hydration levels and thickness. Using both fluorescent and surface plasmon resonance methods, we found that the antifouling behavior of these brushes is strongly dependent on the aforementioned structural changes. Moreover, preconditioning of the brush coatings (incubation at a variable salt concentration or drying) prior to biomolecule interaction may significantly improve the antifouling performance. These results suggest a new simple approach to improve the antifouling behavior of polymer brushes. In addition, the results herein enhance the understanding for improved design of antifouling and bioresponsive brushes employed in biosensor and biomimetic applications.

14.
Macromol Biosci ; 20(3): e1900351, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32045093

RESUMO

Ultra-low fouling and functionalizable coatings represent emerging surface platforms for various analytical and biomedical applications such as those involving examination of cellular interactions in their native environments. Ultra-low fouling surface platforms as advanced interfaces enabling modulation of behavior of living cells via tuning surface physicochemical properties are presented and studied. The state-of-art ultra-low fouling surface-grafted polymer brushes of zwitterionic poly(carboxybetaine acrylamide), nonionic poly(N-(2-hydroxypropyl)methacrylamide), and random copolymers of carboxybetaine methacrylamide (CBMAA) and HPMAA [p(CBMAA-co-HPMAA)] with tunable molar contents of CBMAA and HPMAA are employed. Using a model Huh7 cell line, a systematic study of surface wettability, swelling, and charge effects on the cell growth, shape, and cytoskeleton distribution is performed. This study reveals that ultra-low fouling interfaces with a high content of zwitterionic moieties (>65 mol%) modulate cell behavior in a distinctly different way compared to coatings with a high content of nonionic HPMAA. These differences are attributed mostly to the surface hydration capabilities. The results demonstrate a high potential of carboxybetaine-rich ultra-low fouling surfaces with high hydration capabilities and minimum background signal interferences to create next-generation bioresponsive interfaces for advanced studies of living objects.


Assuntos
Materiais Revestidos Biocompatíveis , Citoesqueleto/metabolismo , Teste de Materiais , Polímeros , Linhagem Celular Tumoral , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Humanos , Polímeros/química , Polímeros/farmacologia , Molhabilidade
15.
J Biomed Mater Res A ; 105(11): 2995-3005, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28646555

RESUMO

Long-term performance of implanted cardiovascular grafts can be ensured if living endothelium overgrows their surface. Surface modifications to implants are therefore being sought that can encourage endothelialization while preventing thrombus formation until the natural endothelium is formed. In the present study, heparin was covalently attached to a fibrin mesh grown from a polyvinyl chloride (PVC) substrate surface by the catalytic action of surface immobilized thrombin on a fibrinogen solution. The coating prevented platelet activation, thrombin generation and clot formation, and reduced inflammatory reactions when exposed to fresh human whole blood circulating in a Chandler loop model. In addition, in vitro seeded human umbilical vein and human saphenous vein endothelial cells showed considerably enhanced attachment and proliferation on the coating. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2995-3005, 2017.


Assuntos
Anticoagulantes/química , Anticoagulantes/farmacologia , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Heparina/química , Heparina/farmacologia , Trombose/prevenção & controle , Coagulação Sanguínea/efeitos dos fármacos , Prótese Vascular/efeitos adversos , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Fibrina/química , Hematócrito , Células Endoteliais da Veia Umbilical Humana , Humanos , Ativação Plaquetária/efeitos dos fármacos , Veia Safena/citologia , Trombose/sangue , Trombose/etiologia
16.
Anal Chem ; 89(6): 3524-3531, 2017 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-28233990

RESUMO

Fouling from complex biological fluids such as blood plasma to biorecognition element (BRE)-functionalized coatings hampers the use of affinity biosensor technologies in medical diagnostics. Here, we report the effects the molecular mechanisms involved in functionalization of low-fouling carboxy-functional coatings have on the BRE capacity and resistance to fouling from blood plasma. The specific mechanisms of EDC/NHS activation of carboxy groups, BRE attachment, and deactivation of residual activated groups on recently developed ultra-low-fouling carboxybetaine polymer and copolymer brushes (pCB) as well as conventional carboxy-terminated oligo(ethylene glycol)-based alkanethiolate self-assembled monolayers (OEG-SAMs) are studied using the polarization modulation infrared reflection/absorption spectroscopy, X-ray photoelectron spectroscopy, and surface plasmon resonance methods. It is shown that the fouling resistance of BRE-functionalized pCB coatings is strongly influenced by a deactivation method affecting the ultra-low-fouling molecular structure of the brush and surface charges. It is revealed that, in contrast to free carboxy-group-terminated OEG-SAMs, only a partial deactivation of EDC/NHS-activated zwitterionic carboxy groups by spontaneous hydrolysis is possible in the pCB brushes. The fouling resistance of activated/BRE-functionalized pCB is shown to be recovered only by covalent attachment of amino acid deactivation agents to residual activated carboxy groups of pCB. The developed deactivation procedure is further combined with ultra-low-fouling brushes of random copolymer carboxybetaine methacrylamide (CBMAA) and N-(2-hydroxypropyl) methacrylamide (HPMAA) with optimized CBMAA content (15%) providing a BRE-functionalized coating with superior fouling resistance over various carboxy-functional low-fouling coatings including homopolymer pCB brushes and OEG-SAMs. The biorecognition capabilities of pHPMAA-CBMAA(15%) are demonstrated via the sensitive label-free detection of a microRNA cancer biomarker (miR-16) in blood plasma.


Assuntos
Materiais Revestidos Biocompatíveis/metabolismo , Polímeros/metabolismo , Materiais Revestidos Biocompatíveis/química , Humanos , Estrutura Molecular , Espectroscopia Fotoeletrônica , Polímeros/síntese química , Polímeros/química
17.
Anal Chem ; 88(21): 10533-10539, 2016 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-27689386

RESUMO

Functional polymer coatings that combine the ability to resist nonspecific fouling from complex media with high biorecognition element (BRE) immobilization capacity represent an emerging class of new functional materials for a number of bioanalytical and biosensor technologies for medical diagnostics, security, and food safety. Here, we report on a random copolymer brush surface - poly(CBMAA-ran-HPMAA) - providing high BRE immobilization capacity while simultaneously exhibiting ultralow-fouling behavior in complex food media. We demonstrate that both the functionalization and fouling resistance capabilities of such copolymer brushes can be tuned by changing the surface contents of the two monomer units: nonionic N-(2-hydroxypropyl) methacrylamide (HPMAA) and carboxy-functional zwitterionic carboxybetaine methacrylamide (CBMAA). It is demonstrated that the resistance to fouling decreases with the surface content of CBMAA; poly(CBMAA-ran-HPMAA) brushes with CBMAA molar content up to 15 mol % maintain excellent resistance to fouling from a variety of homogenized foods (hamburger, cucumber, milk, and lettuce) even after covalent attachment of BREs to carboxy groups of CBMAA. The poly(CBMAA 15 mol %-ran-HPMAA) brushes functionalized with antibodies are demonstrated to exhibit fouling resistance from food samples by up to 3 orders of magnitude better when compared with the widely used low-fouling carboxy-functional oligo(ethylene glycol) (OEG)-based alkanethiolate self-assembled monolayers (AT SAMs) and, furthermore, by up to 2 orders of magnitude better when compared with the most successful ultralow-fouling biorecognition coatings - poly(carboxybetaine acrylamide), poly(CBAA). When model SPR detections of food-borne bacterial pathogens in homogenized foods are used, it is also demonstrated that the antibody-functionalized poly(CBMAA 15 mol %-ran-HPMAA) brush exhibits superior biorecognition properties over the poly(CBAA).


Assuntos
Acrilamidas/química , Resinas Acrílicas/química , Incrustação Biológica/prevenção & controle , Inocuidade dos Alimentos/métodos , Resinas Acrílicas/síntese química , Anticorpos/química , Escherichia coli/imunologia , Alimentos , Ouro/química , Nanopartículas/química , Salmonella typhimurium/imunologia , Molhabilidade
18.
PLoS One ; 10(5): e0125484, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25945799

RESUMO

In a typical cell culture system, growth factors immobilized on the cell culture surfaces can serve as a reservoir of bio-signaling molecules, without the need to supplement them additionally into the culture medium. In this paper, we report on the fabrication of albumin/heparin (Alb/Hep) assemblies for controlled binding of basic fibroblast growth factor (FGF-2). The surfaces were constructed by layer-by-layer adsorption of polyelectrolytes albumin and heparin and were subsequently stabilized by covalent crosslinking with glutaraldehyde. An analysis of the surface morphology by atomic force microscopy showed that two Alb/Hep bilayers are required to cover the surface of substrate. The formation of the Alb/Hep assemblies was monitored by the surface plasmon resonance (SPR), the infrared multiinternal reflection spectroscopy (FTIR MIRS) and UV/VIS spectroscopy. The adsorption of FGF-2 on the cross-linked Alb/Hep was followed by SPR. The results revealed that FGF-2 binds to the Alb/Hep assembly in a dose and time-dependent manner up to the surface concentration of 120 ng/cm(2). The bioactivity of the adsorbed FGF-2 was assessed in experiments in vitro, using calf pulmonary arterial endothelial cells (CPAE). CPAE cells could attach and proliferate on Alb/Hep surfaces. The adsorbed FGF-2 was bioactive and stimulated both the proliferation and the differentiation of CPAE cells. The improvement was more pronounced at a lower FGF-2 surface concentration (30 ng/cm(2)) than on surfaces with a higher concentration of FGF-2 (120 ng/cm(2)).


Assuntos
Albuminas/farmacologia , Técnicas de Cultura de Células/métodos , Células Endoteliais/citologia , Fator 2 de Crescimento de Fibroblastos/farmacologia , Heparina/farmacologia , Animais , Bovinos , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Meios de Cultura/química , Fator 2 de Crescimento de Fibroblastos/metabolismo , Humanos , Microscopia de Força Atômica , Ligação Proteica
19.
Beilstein J Nanotechnol ; 6: 617-631, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25821702

RESUMO

Composite materials based on a titanium support and a thin, alginate hydrogel could be used in bone tissue engineering as a scaffold material that provides biologically active molecules. The main objective of this contribution is to characterize the activation and the functionalization of titanium surfaces by the covalent immobilization of anchoring layers of self-assembled bisphosphonate neridronate monolayers and polymer films of 3-aminopropyltriethoxysilane and biomimetic poly(dopamine). These were further used to bind a bio-functional alginate coating. The success of the titanium surface activation, anchoring layer formation and alginate immobilization, as well as the stability upon immersion under physiological-like conditions, are demonstrated by different surface sensitive techniques such as spectroscopic ellipsometry, infrared reflection-absorption spectroscopy and X-ray photoelectron spectroscopy. The changes in morphology and the established continuity of the layers are examined by scanning electron microscopy, surface profilometry and atomic force microscopy. The changes in hydrophilicity after each modification step are further examined by contact angle goniometry.

20.
Tissue Eng Part A ; 20(15-16): 2253-64, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24606163

RESUMO

Vascular surgery for atherosclerosis is confronted by the lack of a suitable bypass material. Tissue engineering strives to produce bio-artificial conduits to provide resistance to thrombosis. The objectives of our study were to culture endothelial cells (EC) on composite assemblies of extracellular matrix proteins, and to evaluate the cellular phenotype under flow. Cell-adhesive assemblies were fabricated on glass slides as combinations of collagen (Co), laminin (LM), and fibronectin (FN), resulting in three samples: Co, Co/LM, and Co/FN. Surface topography, roughness, and wettability were determined. Human saphenous vein EC were harvested from cardiac patients, cultured on the assemblies and submitted to laminar shear stress (SS) of 12 dyn/cm(2) for 40, 80, and 120 min. Cell retention was assessed and qRT-PCR of adhesion genes (VE-cadherin, vinculin, KDR, CD-31 or PECAM-1, ß1-integrins) and metabolic genes (t-PA, NF-κB, eNOS and MMP-1) was performed. Quantitative immunofluorescence of VE cadherin, vinculin, KDR, and vonWillebrand factor was performed after 2 and 6 h of flow. Static samples were excluded from shearing. The cells reached confluence with similar growth curves. The cells on Co/LM and Co/FN were resistant to flow up to 120 min but minor desquamation occurred on Co corresponding with temporary downregulation of VE cadherin and vinculin-mRNA and decreased fluorescence of vinculin. The cells seeded on Co/LM initially more upregulated vinculin-mRNA and also the inflammatory factor NF-κB, and the cells plated on Co/FN changed the expression profile minimally in comparison with the static control. Fluorescence of VE cadherin and vonWillebrand factor was enhanced on Co/FN. The cells cultured on Co/LM and Co/FN increased the vinculin fluorescence and expressed more VE cadherin and KDR-mRNA than the cells on Co. The cells plated on Co/FN upregulated the mRNA of VE cadherin, CD-31, and MMP 1 to a greater extent than the cells on Co/LM and they enhanced the fluorescence of VE cadherin, KDR, and vonWillebrand factor. Some of these changes sustained up to 6 h of flow, as confirmed by immunofluorescence. Combined matrices Co/LM and Co/FN seem to be more suitable for EC seeding and retention under flow. Moreover, Co/FN matrix promoted slightly more favorable cellular phenotype than Co/LM under SS of 2-6 h.


Assuntos
Células Endoteliais/metabolismo , Proteínas da Matriz Extracelular/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Resistência ao Cisalhamento , Estresse Mecânico , Animais , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Imunofluorescência , Perfilação da Expressão Gênica , Humanos , Camundongos , Ratos , Veia Safena/citologia , Ressonância de Plasmônio de Superfície , Fatores de Tempo , Molhabilidade
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