INTRODUCTION: Microarray-based gene expression profiling represents a major breakthrough for understanding the molecular complexity of breast cancer. cDNA expression profiles cannot detect changes in activities that arise from post-translational modifications, however, and therefore do not provide a complete picture of all biologically important changes that occur in tumors. Additional opportunities to identify and/or validate molecular signatures of breast carcinomas are provided by proteomic approaches. Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) offers high-throughput protein profiling, leading to extraction of protein array data, calling for effective and appropriate use of bioinformatics and statistical tools. METHODS: Whole tissue lysates of 105 breast carcinomas were analyzed on IMAC 30 ProteinChip Arrays (Bio-Rad, Hercules, CA, USA) using the ProteinChip Reader Model PBS IIc (Bio-Rad) and Ciphergen ProteinChip software (Bio-Rad, Hercules, CA, USA). Cluster analysis of protein spectra was performed to identify protein patterns potentially related to established clinicopathological variables and/or tumor markers. RESULTS: Unsupervised hierarchical clustering of 130 peaks detected in spectra from breast cancer tissue lysates provided six clusters of peaks and five groups of patients differing significantly in tumor type, nuclear grade, presence of hormonal receptors, mucin 1 and cytokeratin 5/6 or cytokeratin 14. These tumor groups resembled closely luminal types A and B, basal and HER2-like carcinomas. CONCLUSION: Our results show similar clustering of tumors to those provided by cDNA expression profiles of breast carcinomas. This fact testifies the validity of the SELDI-TOF MS proteomic approach in such a type of study. As SELDI-TOF MS provides different information from cDNA expression profiles, the results suggest the technique's potential to supplement and expand our knowledge of breast cancer, to identify novel biomarkers and to produce clinically useful classifications of breast carcinomas.

• MeSH
• Models, Biological MeSH
• Protein Array Analysis methods   MeSH
• Molecular Diagnostic Techniques MeSH
• Financing, Organized MeSH
• DNA, Complementary metabolism   MeSH
• Humans MeSH
• Biomarkers, Tumor MeSH
• Breast Neoplasms genetics   metabolism   MeSH
• Protein Processing, Post-Translational MeSH
• Proteomics methods   MeSH
• Gene Expression Regulation MeSH
• Cluster Analysis MeSH
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods   MeSH
• Gene Expression Profiling MeSH
• Computational Biology methods   MeSH
• Check Tag
• Humans MeSH
• Female MeSH

The principal focus of this work is the in-depth analysis of the biological efficiency of inorganic calcium-filled bacterial cellulose (BC) based hydrogel scaffolds for their future use in bone tissue engineering/bioengineering. Inorganic calcium was filled in the form of calcium phosphate (β-tri calcium phosphate (β-TCP) and hydroxyapatite (HA)) and calcium carbonate (CaCO₃). The additional calcium, CaCO₃ was incorporated following in vitro bio-mineralization. Cell viability study was performed with the extracts of BC based hydrogel scaffolds: BC-PVP, BC-CMC; BC-PVP-β-TCP/HA, BC-CMC-β-TCP/HA and BC-PVP-β-TCP/HA-CaCO₃, BC-CMC-β-TCP/HA-CaCO₃; respectively. The biocompatibility study was performed with two different cell lines, i.e., human fibroblasts, Lep-3 and mouse bone explant cells. Each hydrogel scaffold has facilitated notable growth and proliferation in presence of these two cell types. Nevertheless, the percentage of DNA strand breaks was higher when cells were treated with BC-CMC based scaffolds i.e., BC-CMC-β-TCP/HA and BC-CMC-β-TCP/HA-CaCO₃. On the other hand, the apoptosis of human fibroblasts, Lep-3 was insignificant in BC-PVP-β-TCP/HA. The scanning electron microscopy confirmed the efficient adhesion and growth of Lep-3 cells throughout the surface of BC-PVP and BC-PVP-β-TCP/HA. Hence, among all inorganic calcium filled hydrogel scaffolds, 'BC-PVP-β-TCP/HA' was recommended as an efficient tissue engineering scaffold which could facilitate the musculoskeletal (i.e., bone tissue) engineering/bioengineering.

• MeSH
• Cellulose chemistry   MeSH
• Hydrogels chemistry   MeSH
• Durapatite chemistry   MeSH
• Bone and Bones cytology   MeSH
• Humans MeSH
• Mice MeSH
• Tissue Engineering methods   MeSH
• Tissue Scaffolds chemistry   MeSH
• Calcium chemistry   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Fabrication of porous and biologically inspired biomaterials that mimic the formation of microstructural structures of nacre in the form of calcite (CaCO3) and evaluation of the biocompatibility of such organic-inorganic composite scaffold for bone tissue engineering, are focus of this paper. Nacre's self-assembly characteristics are concerned about the development of calcite filled biomineralized scaffold following the nature based biomineralization process and biomimetic applications. The PVP-CMC hydrogel film, comprised of PVP:0.2, CMC:0.8, PEG:1.0, Agar:2.0, Glycerene:1.0 and water:95.0 w/v%; acts as catalyst and template for the nucleation and growth of the inorganic CaCO3 within the scaffold. The PVP-CMC hydrogel (in the dry state) was immersed in ionic solutions (g/100 ml) of Na2CO3 and CaCl2·H2O in different concentrations sets i.e. Set-1: 10.50/14.70; Set-2: 5.25/7.35; Set-3: 4.20/5.88; Set-4: 2.10/2.94; Set-5: 1.05/1.47, Set-6: 0.55/0.55 for 90 min. As a result, "PVP-CMC-CaCO3" hydrogel scaffold was fabricated having bio-inspired structural and functional properties. Cell proliferation and cell viability were examined until 7 days in the presence of "PVP-CMC-CaCO3" scaffolds using permanent cell lines MG63 (human osteosarcoma), L929 (murine fibroblasts) as well as cultures from mouse bone explants (CC-MBE), confirmed that the said hydrogel scaffolds are biocompatible. But, from mechanical strength as well as biocompatibility point of view, scaffolds prepared in Set-1 to 3 ionic solutions were superior. In conclusion, these three calcite filled hydrogel scaffolds are recommended and can be used for osseointegration.

• MeSH
• Biocompatible Materials chemistry   MeSH
• Cell Line MeSH
• X-Ray Diffraction MeSH
• Hydrogels chemistry   MeSH
• Microscopy, Electron, Scanning MeSH
• Mice MeSH
• Osseointegration physiology   MeSH
• Spectroscopy, Fourier Transform Infrared MeSH
• Tissue Engineering methods   MeSH
• Tissue Scaffolds chemistry   MeSH
• Cell Survival physiology   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

In a biological system, nanoparticles (NPs) may interact with biomolecules. Specifically, the adsorption of proteins on the nanoparticle surface may influence both the nanoparticles' and proteins' overall bio-reactivity. Nevertheless, our knowledge of the biocompatibility and risk of exposure to nanomaterials is limited. Here, in vitro and ex ovo biocompatibility of naturally based crosslinked freeze-dried 3D porous collagen/chitosan scaffolds, modified with thermostable fibroblast growth factor 2 (FGF2-STAB®), to enhance healing and selenium nanoparticles (SeNPs) to provide antibacterial activity, were evaluated. Biocompatibility and cytotoxicity were tested in vitro using normal human dermal fibroblasts (NHDF) with scaffolds and SeNPs and FGF2-STAB® solutions. Metabolic activity assays indicated an antagonistic effect of SeNPs and FGF2-STAB® at high concentrations of SeNPs. The half-maximal inhibitory concentration (IC50) of SeNPs for NHDF was 18.9 µg/ml and IC80 was 5.6 µg/ml. The angiogenic properties of the scaffolds were monitored ex ovo using a chick chorioallantoic membrane (CAM) assay and the cytotoxicity of SeNPs over IC80 value was confirmed. Furthermore, the positive effect of FGF2-STAB® at very low concentrations (0.01 µg/ml) on NHDF metabolic activity was observed. Based on detailed in vitro testing, the optimal concentrations of additives in the scaffolds were determined, specifically 1 µg/ml of FGF2-STAB® and 1 µg/ml of SeNPs. The scaffolds were further subjected to antimicrobial tests, where an increase in selenium concentration in the collagen/chitosan scaffolds increased the antibacterial activity. This work highlights the antimicrobial ability and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB® and SeNPs. Moreover, we suggest that these sponges could be used as scaffolds for growing cells in systems with low mechanical loading in tissue engineering, especially in dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration. Due to their antimicrobial properties, these scaffolds are also highly promising for tissue replacement requiring the prevention of infection.

• MeSH
• Anti-Bacterial Agents MeSH
• Biocompatible Materials pharmacology   MeSH
• Cell Line MeSH
• Chitosan pharmacology   MeSH
• Fibroblast Growth Factor 2 pharmacology   MeSH
• Fibroblasts drug effects   MeSH
• Wound Healing MeSH
• Collagen pharmacology   MeSH
• Humans MeSH
• Nanoparticles chemistry   therapeutic use   MeSH
• Porosity MeSH
• Selenium chemistry   pharmacology   MeSH
• Materials Testing MeSH
• Tissue Engineering methods   MeSH
• Tissue Scaffolds * MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Repairing and regenerating damaged tissues or organs, and restoring their functioning has been the ultimate aim of medical innovations. 'Reviving healthcare' blends tissue engineering with alternative techniques such as hydrogels, which have emerged as vital tools in modern medicine. Additive manufacturing (AM) is a practical manufacturing revolution that uses building strategies like molding as a viable solution for precise hydrogel manufacturing. Recent advances in this technology have led to the successful manufacturing of hydrogels with enhanced reproducibility, accuracy, precision, and ease of fabrication. Hydrogels continue to metamorphose as the vital compatible bio-ink matrix for AM. AM hydrogels have paved the way for complex 3D/4D hydrogels that can be loaded with drugs or cells. Bio-mimicking 3D cell cultures designed via hydrogel-based AM is a groundbreaking in-vivo assessment tool in biomedical trials. This brief review focuses on preparations and applications of additively manufactured hydrogels in the biomedical spectrum, such as targeted drug delivery, 3D-cell culture, numerous regenerative strategies, biosensing, bioprinting, and cancer therapies. Prevalent AM techniques like extrusion, inkjet, digital light processing, and stereo-lithography have been explored with their setup and methodology to yield functional hydrogels. The perspectives, limitations, and the possible prospects of AM hydrogels have been critically examined in this study.

• MeSH
• Printing, Three-Dimensional MeSH
• Bioprinting methods   MeSH
• Cell Culture Techniques MeSH
• Hydrogels * chemistry   MeSH
• Drug Delivery Systems MeSH
• Humans MeSH
• Cell Culture Techniques, Three Dimensional methods   MeSH
• Tissue Engineering * methods   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

Nanoparticles functionalized with specific biological recognition molecules play a major role for sensor response enhancement in surface plasmon resonance (SPR) based biosensors. The functionalization procedure of such nanoparticles is crucial, since it influences their interactions with the environment and determines their applicability to biomolecular detection in complex matrices. In this work we show how the ζ-potential (Zpot) of bio-functionalized gold spherical NPs (Bio-NPs) is related to the SPR sensor response enhancement of an immune-sandwich-assay for the detection of the carcinoembryonic antigen (CEA), a cancer marker for colorectal carcinomas. In particular, we prepare bio-functional nanoparticles by varying the amount of peptide (either streptavidin or antibody against CEA) bound on their surface. Specific and non-specific sensor responses, reproducibility, and colloidal stability of those bio-functional nanoparticles are measured via SPR and compared to ζ-potential values. Those parameters are first measured in buffer solution, then measured again when the surface of the biosensor is exposed to blood plasma, and finally when the nanoparticles are immersed in blood plasma and flowed overnight on the biosensor. We found that ζ-potential values can guide the design of bio-functional NPs with improved binding efficiency and reduced non-specific sensor response, suitable reproducibility and colloidal stability, even in complex matrixes like blood plasma.

• MeSH
• Hematologic Tests methods   MeSH
• Carcinoembryonic Antigen * blood   MeSH
• Plasma chemistry   MeSH
• Humans MeSH
• Biomarkers, Tumor analysis   MeSH
• Nanoparticles analysis   MeSH
• Peptides analysis   MeSH
• Surface Plasmon Resonance methods   MeSH
• Gold analysis   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Bio-artificial polymeric systems are a new class of polymeric constituents based on blends of synthetic and natural polymers, designed with the purpose of producing new materials that exhibit enhanced properties with respect to the individual components. In this frame, a combination of polyvinyl alcohol (PVA) and chitosan, blended with a widely used antibiotic, sodium ampicillin, has been developed showing a moderate behavior in terms of antibacterial properties. Thus, aqueous solutions of PVA at 1 wt.% were mixed with acid solutions of chitosan at 1 wt.%, followed by adding ampicillin ranging from 0.3 to 1.0 wt.% related to the total amount of the polymers. The prepared bio-artificial polymeric system was characterized by FTIR, SEM, DSC, contact angle measurements, antibacterial activity against Staphylococcus aureus and Escherichia coli and antibiotic release studies. The statistical significance of the antibacterial activity was determined using a multifactorial analysis of variance with ρ < 0.05 (ANOVA). The characterization techniques did not show alterations in the ampicillin structure and the interactions with polymers were limited to intermolecular forces. Therefore, the antibiotic was efficiently released from the matrix and its antibacterial activity was preserved. The system disclosed moderate antibacterial activity against bacterial strains without adding a high antibiotic concentration. The findings of this study suggest that the system may be effective against healthcare-associated infections, a promising view in the design of novel antimicrobial biomaterials potentially suitable for tissue engineering applications.

• MeSH
• Ampicillin * chemistry   pharmacology   MeSH
• Anti-Bacterial Agents * chemical synthesis   chemistry   pharmacology   MeSH
• Chitosan * chemistry   pharmacology   MeSH
• Escherichia coli growth & development   MeSH
• Polyvinyl Alcohol * chemistry   pharmacology   MeSH
• Staphylococcus aureus growth & development   MeSH
• Publication type
• Journal Article MeSH

The purpose of this study was to compare, by means of in vitro cultivation technique, five marketed brands of wound covers used in the treatment of burns and other skin defects (Biobrane(®), Suprathel(®), Veloderm(®), Xe-Derma(®), and Xenoderm(®)) for their ability to stimulate the keratinocyte growth, stratification, and differentiation. In three independent experiments, human keratinocytes were grown on the tested covers in organotypic cultures by the 3T3 feeder layer technique. Vertical paraffin sections of the wound covers with keratinocytes were processed using hematoxylin-eosin staining and immunostaining for involucrin. Keratinocyte populations on the dressings were assessed for (1) number of keratinocyte strata (primary variable), (2) quantitative growth, (3) thickness of the keratinocyte layer, and (4) cell differentiation. The Xe-Derma wound cover provided the best support to keratinocyte proliferation and stratification, with the number of keratinocyte strata significantly (p < 0.05) higher in comparison to all products studied, except Xenoderm. However, in contrast to Xe-Derma, Xenoderm did not significantly differ from the other dressings. The results of this in vitro study show that the brands based on porcine dermal matrix possess the strongest effect on keratinocyte proliferation and stratification. The distinctive position of Xe-Derma may be related to its composition, where natural dermal fibers form a smooth surface, similar to the basement membrane. Furthermore, the results indicate that in vitro evaluation of effects on epithelial growth may accelerate the development of new bio-engineering-based wound covers.

• Publication type
• Journal Article MeSH

INTRODUCTION: The presence of microbial invasion of the amniotic cavity (MIAC) and histological chorioamnionitis (HCA) is associated with adverse neonatal outcomes in pregnancies complicated by preterm prelabor rupture of membranes (pPROM). Therefore, there is an urgent need to identify new biomarkers revealing these conditions. The objective of this study is to identify possible biomarkers and their underlying biofunctions in pPROM pregnancies with and without MIAC and HCA. METHODS: A total of 72 women with pPROM were recruited. Only women with both MIAC and HCA (n = 19) and all women without these complications (n = 19) having the same range of gestational ages at sampling were included in the study. Samples of amniotic fluid were obtained by transabdominal amniocentesis, processed and analyzed using quantitative shotgun proteomics. Ingenuity pathway analysis was used to identify molecular networks that involve altered proteins. RESULTS: Network interaction identified by ingenuity pathway analysis revealed immunological disease and the inflammatory response as the top functions and disease associated with pPROM in the presence of MIAC and HCA. The proteins involved in these pathways were significantly altered between the groups with and without the presence of both MIAC and HCA. Proteins involved included histones H3, H4, H2B, cathelicidin antimicrobial peptide, myeloperoxidase, neutrophil gelatinase-associated lipocalin, matrix metalloproteinase-9, peptidoglycan recognition protein-1 and neutrophil defensin 1, all of which were found to be up-regulated in the presence of MIAC and HCA. CONCLUSION: Bioinformatic analysis of proteomics data allowed us to project likely biomolecular pathology resulting in pPROM complicated by MIAC and HCA. As inflammation is not a homogeneous phenomenon, we provide evidence for oxidative-stress-associated DNA damage and biomarkers of reactive oxygen species generation as factors associated with inflammation and proteolysis.

• MeSH
• Biomarkers metabolism   MeSH
• Chorioamnionitis immunology   metabolism   MeSH
• Adult MeSH
• Histones metabolism   MeSH
• Cohort Studies MeSH
• Humans MeSH
• Inflammation Mediators metabolism   MeSH
• Metabolic Networks and Pathways MeSH
• Young Adult MeSH
• Infant, Newborn MeSH
• Oxidative Stress MeSH
• Fetal Membranes, Premature Rupture immunology   metabolism   MeSH
• Proteomics MeSH
• Case-Control Studies MeSH
• Pregnancy MeSH
• Computational Biology MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Infant, Newborn MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• European Union MeSH
• Research MeSH
• Publication type
• Abstracts MeSH
• Geographicals
• Europe MeSH

• Conspectus
• Veřejné zdraví a hygiena
• NML Fields
• veřejné zdravotnictví
• politologie, politika, zdravotní politika

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• Commission of the European Communities. Biomedical and Health Research Programme Authority