scholarly journals Soy Protein-Based Composite Hydrogels: Physico-Chemical Characterization and In Vitro Cytocompatibility

Polymers ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 1159 ◽  
Author(s):  
Samira Tansaz ◽  
Raminder Singh ◽  
Iwona Cicha ◽  
Aldo Boccaccini
Keyword(s):  
Soft Tissue ◽  
Metabolic Activity ◽  
Soy Protein ◽  
Colorimetric Assay ◽  
Umbilical Vein ◽  
Chemical Characterization ◽  
Dermal Fibroblasts ◽  
Water Soluble ◽  
Composite Hydrogels

Novel composite hydrogels based on the combination of alginate (Alg), soy protein isolate (SPI) and bioactive glass (BG) nanoparticles were developed for soft tissue engineering. Human umbilical vein endothelial cells (HUVEC) and normal human dermal fibroblasts were cultivated on hydrogels for 7, 14 and 21 days. Cell morphology was visualized using fluorescent staining at Days 7 and 14 for fibroblast cells and Days 14 and 21 for HUVEC. Metabolic activity of cells was analyzed using a colorimetric assay (water soluble tetrazolium (WST) assay). Compared to pure Alg, Alg/SPI and Alg/SPI/BG provided superior surfaces for both types of cells, supporting their attachment, growth, spreading and metabolic activity. Fibroblasts showed better colonization and growth on Alg/SPI/BG hydrogels compared to Alg/SPI hydrogels. The results indicate that such novel composite hydrogels might find applications in soft tissue regeneration.

scholarly journals Mutual influence of selenium nanoparticles and FGF2-STAB® on biocompatible properties of collagen/chitosan 3D scaffolds: in vitro and ex ovo evaluation

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Johana Muchová ◽  
Vanessa Hearnden ◽  
Lenka Michlovská ◽  
Lucie Vištejnová ◽  
Anna Zavaďáková ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Metabolic Activity ◽  
Mutual Influence ◽  
Chorioallantoic Membrane ◽  
Antagonistic Effect ◽  
Dermal Fibroblasts ◽  
Selenium Nanoparticles ◽  
Freeze Dried ◽  
Chitosan Scaffolds

AbstractIn 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.

scholarly journals Single-Cell Receptor Quantification of an In Vitro Coculture Angiogenesis Model Reveals VEGFR, NRP1, Tie2, and PDGFR Regulation and Endothelial Heterogeneity

Processes ◽  
2019 ◽  
Vol 7 (6) ◽  
pp. 356 ◽  
Author(s):  
Si Chen ◽  
P. I. Imoukhuede
Keyword(s):  
Systems Biology ◽  
Growth Factor ◽  
Plasma Membranes ◽  
Temporal Dynamics ◽  
Ex Vivo ◽  
Umbilical Vein ◽  
Tube Formation ◽  
Dermal Fibroblasts ◽  
Steady Increase

Angiogenesis, the formation of new blood vessels from pre-existing ones, is essential for both normal development and numerous pathologies. Systems biology has offered a unique approach to study angiogenesis by profiling tyrosine kinase receptors (RTKs) that regulate angiogenic processes and computationally modeling RTK signaling pathways. Historically, this systems biology approach has been applied on ex vivo angiogenesis assays, however, these assays are difficult to quantify and limited in their potential of temporal analysis. In this study, we adopted a simple two-dimensional angiogenesis assay comprised of human umbilical vein endothelial cells (HUVECs) and human dermal fibroblasts (HDFs) and examined temporal dynamics of a panel of six RTKs and cell heterogeneity up to 17 days. We observed ~2700 VEGFR1 (vascular endothelial growth factor receptor 1) per cell on 24-h-old cocultured HDF plasma membranes, which do not express VEGFR when cultured alone. We observed 4000–8100 VEGFR2 per cell on cocultured HUVEC plasma membranes throughout endothelial tube formation. We showed steady increase of platelet-derived growth factor receptors (PDGFRs) on cocultured HDF plasma membranes, and more interestingly, 1900–2900 PDGFRβ per plasma membrane were found on HUVECs within the first six hours of coculturing. These quantitative findings will offer us insights into molecular regulation during angiogenesis and help assess in vitro tube formation models and their physiological relevance.

scholarly journals The role of integrins in the maintenance of endothelial monolayer integrity.

1991 ◽  
Vol 112 (3) ◽  
pp. 479-490 ◽  
Author(s):  
M G Lampugnani ◽  
M Resnati ◽  
E Dejana ◽  
P C Marchisio
Keyword(s):  
Umbilical Vein ◽  
Dermal Fibroblasts ◽  
Collagen Type Iv ◽  
Cell Contact ◽  
Endothelial Monolayer ◽  
Integrin Receptors ◽  
Endothelial Lining ◽  
Type Iv ◽  
Cell Cell

This paper shows that, in confluent human umbilical vein endothelial cell (EC) monolayers, the integrin heterodimers alpha 2 beta 1 and alpha 5 beta 1, but not other members of the beta 1 subfamily, are located at cell-cell contact borders and not at cellular free edges. Also the alpha v chain, but not its most common partner beta 3, that is widely expressed in EC cell-matrix junctions, is found at cell-cell borders. In EC monolayers, the putative ligands of alpha 2 beta 1 and alpha 5 beta 1 receptors, i.e., laminin, collagen type IV, and fibronectin, are also organized in strands corresponding to cell-cell borders. The location of the above integrin receptors is not an artifact of in vitro culture since it has been noted also in explanted islets of the native umbilical vein endothelium. The integrins alpha 2 beta 1 and alpha 5 beta 1 play a role in the maintenance of endothelial monolayer continuity in vitro. Indeed, specific antibodies to alpha 2 beta 1, alpha 5 beta 1, and the synthetic peptide GRGDSP alter its continuity without any initial cell detachment. Moreover, antibodies to alpha 5 beta 1 increase the permeation of macromolecules across confluent EC monolayers. In contrast beta 3 antibodies were ineffective. It is suggested that the relocation of integrins to cell-cell borders is a feature of cells programmed to form polarized monolayers since integrins have a different distribution in nonpolar confluent dermal fibroblasts. The conclusion is that some members of the integrin superfamily collaborate with other intercellular molecules to form lateral junctions and to control both the monolayer integrity and the permeability properties of the vascular endothelial lining. This also suggest that integrins are adhesion molecules provided with a unique biochemical adaptability to different biological functions.

scholarly journals Pharmacologic Characterization of a Kinetic In Vitro Human Co-Culture Angiogenesis Model Using Clinically Relevant Compounds

2013 ◽  
Vol 18 (10) ◽  
pp. 1234-1245 ◽  
Author(s):  
Ashley Wolfe ◽  
Belinda O’Clair ◽  
Vincent E. Groppi ◽  
Dyke P. McEwen
Keyword(s):  
Growth Factor ◽  
Umbilical Vein ◽  
Dermal Fibroblasts ◽  
Tube Length ◽  
Endothelial Cell Proliferation ◽  
Discovery Research ◽  
Drug Discovery Research ◽  
Angiogenesis Model

Angiogenesis, the formation of new vessels from preexisting vessels, involves multiple cell types acting in concert to cause endothelial cell proliferation, migration, and differentiation into microvascular arrays. Under pathologic conditions, microenvironment changes result in altered blood vessel production. Historically, in vitro angiogenesis assays study individual aspects of the process and tend to be variable, difficult to quantify, and limited in clinical relevance. Here, we describe a kinetic, quantitative, co-culture angiogenesis model and demonstrate its relevance to in vivo pharmacology. Similar to in vivo angiogenesis, a co-culture of human umbilical vein endothelial cells with normal human dermal fibroblasts remains sensitive to multiple cytokines, resulting in a concentration-dependent stimulation of tube formation over time. Treatment with axitinib, a selective vascular endothelial growth factor (VEGF) antagonist, inhibited VEGF-mediated tube length and branch point formation and was selective for inhibiting VEGF over basic fibroblast growth factor (bFGF), similar to previous studies. Conversely, an FGFR-1 selective compound, PD-161570, was more potent at inhibiting bFGF-mediated angiogenesis. These results demonstrate the cytokine dynamics, selective pharmacology, and translational application of this model system. Finally, combining quantitative angiogenic biology with kinetic, live-content imaging highlights the importance of using validated in vitro models in drug discovery research.

scholarly journals Mechanical stimulation of human dermal fibroblasts regulates pro-inflammatory cytokines: potential insight into soft tissue manual therapies

2020 ◽  
Author(s):  
Aric Anloague ◽  
Aaron Mahoney ◽  
Oladipupo Ogunbekun ◽  
William R. Thompson ◽  
Bryan Larsen ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Inflammatory Cytokines ◽  
Mechanical Stimulation ◽  
Mechanical Strain ◽  
Dermal Fibroblasts ◽  
Manual Therapies ◽  
Human Dermal Fibroblasts ◽  
Stimulation Of ◽  
Pro Inflammatory Cytokines

Abstract Objective Soft tissue manual therapies are commonly utilized by osteopathic physicians, chiropractors, physical therapists and massage therapists. These techniques are predicated on subjecting tissues to biophysical mechanical stimulation but the cellular and molecular mechanism(s) mediating these effects are poorly understood. A series of previous studies established an in vitro model system for examining mechanical stimulation of dermal fibroblasts and established that repetitive strain, intended to mimic overuse injury, induces the secretion of numerous pro-inflammatory cytokines. Moreover, mechanical strain intended to mimic soft tissue manual therapy reduces strain-induced secretion of pro-inflammatory cytokines. Here, we sought to partially confirm and extend these reports and provide independent corroboration of prior results. Results Using cultures of primary human dermal fibroblasts, we confirm mechanical forces intended to mimic repetitive motion strain increases levels of IL-6 and that mechanical strain intended to mimic therapeutic soft tissue stimulation reduces IL-6 levels. We also extend the prior work, reporting that therapy-like mechanical stimulation reduces levels of IL-8. Although there are important limitations to this experimental model, these findings provide supportive evidence that therapeutic soft tissue massage may reduce inflammation. Future work is required to address these open questions and advance the mechanistic understanding of therapeutic soft tissue stimulation.

scholarly journals Anti-Inflammatory Effect of Cherry Extract Loaded in Polymeric Nanoparticles: Relevance of Particle Internalization in Endothelial Cells

Pharmaceutics ◽  
2019 ◽  
Vol 11 (10) ◽  
pp. 500 ◽  
Author(s):  
Denise Beconcini ◽  
Francesca Felice ◽  
Ylenia Zambito ◽  
Angela Fabiano ◽  
Anna Maria Piras ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Prunus Avium ◽  
Umbilical Vein ◽  
Water Soluble ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tetrazolium Salt ◽  
Tnf Α ◽  
Anti Inflammatory ◽  
Inflammatory Effect

This study aimed at evaluating the anti-inflammatory effect of natural cherry extract (CE), either free or encapsulated in nanoparticles (NPs) based on chitosan derivatives (Ch-der) or poly(lactic-co-glycolic acid) (PLGA), on human umbilical vein endothelial cells (HUVEC). CE from Prunus avium L. was characterized for total polyphenols, flavonoids, and anthocyanins content. CE and CE-loaded NP cytotoxicity and protective effect on lipopolysaccharide (LPS)-stressed HUVEC were tested by water-soluble tetrazolium salt (WST-1) assay. Pro- and anti-inflammatory cytokines (TNF-α, IL-6, IL-10, and PGE2) released by HUVEC were quantified by enzyme-linked immunosorbent assay (ELISA). All NP types were internalized into HUVEC after 2 h incubation and promoted the anti-inflammatory effect of free CE at the concentration of 2 µg gallic acid equivalents (GAE)/mL. CE-loaded Ch-der NPs showed the highest in vitro uptake and anti-inflammatory activity, blunting the secretion of IL-6, TNF-α, and PGE2 cytokines. Moreover, all NPs reduced the production of nitric oxide and NLRP3 inflammasome, and had a stronger anti-inflammatory effect than the major corticosteroid dexamethasone. In particular, the results demonstrate that natural CE protects endothelial cells from inflammatory stress when encapsulated in NPs based on quaternary ammonium chitosan. The CE beneficial effects were directly related with in vitro internalization of CE-loaded NPs.

Effects of Cola-Flavored Beverages and Caffeine on Streptococcus mutans Biofilm Formation and Metabolic Activity

2017 ◽  
Vol 41 (4) ◽  
pp. 294-299 ◽  
Author(s):  
Roger P Dotsey ◽  
Elizabeth A S Moser ◽  
George J Eckert ◽  
Richard L Gregory
Keyword(s):  
Streptococcus Mutans ◽  
Metabolic Activity ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Bacterial Biofilm ◽  
Water Soluble ◽  
High Fructose Corn Syrup ◽  
Biofilm Growth ◽  
High Fructose

Objective: To examine the effects of cola-flavored beverages and caffeine on growth and metabolism of Streptococcus mutans biofilm. This study was designed to determine if carbonated beverages or caffeine can increase S. mutans growth and biofilm formation and metabolic activity in vitro, potentially leading to increased S. mutans-associated cariogenicity in children that consume them. Study Design: Six different cola-flavored products, plus pure caffeine, and pure high fructose corn syrup (HFCS), at different concentrations similar to those in the beverages were tested. A 16-hour culture of S. mutans was treated with different dilutions in bacteriological media. To test for the effect on biofilm formation, the biofilm was stained with crystal violet. The absorbance was determined to evaluate biofilm growth. Biofilm metabolic activity was measured based on biofilm having the ability to reduce XTT to a water-soluble orange compound. Results: The inclusion of HFCS in the beverages, as well as pure HFCS, significantly enhanced bacterial biofilm formation and metabolic activity. Pure caffeine and the presence of caffeine in beverages did not significantly increase biofilm formation, but pure caffeine significantly increased metabolism, and Diet Coke had significantly greater metabolic activity than Caffeine-Free Diet Coke. Conclusions: HFCS increases both the biofilm formation and metabolism of S. mutans, and caffeine in some cases increases metabolism of S. mutans.

A Bivalent Role of Genistein in Sprouting Angiogenesis

Planta Medica ◽  
2018 ◽  
Vol 84 (09/10) ◽  
pp. 653-661 ◽  
Author(s):  
Sarah Berndt ◽  
Mark Issa ◽  
Gilles Carpentier ◽  
Muriel Cuendet
Keyword(s):  
Endothelial Cell ◽  
Metabolic Activity ◽  
Umbilical Vein ◽  
Cancer Chemoprevention ◽  
Tube Formation ◽  
Antiangiogenic Effect ◽  
Human Umbilical Vein ◽  
Sprouting Angiogenesis ◽  
Genistein Treatment

AbstractThe effects of genistein on angiogenesis remain poorly understood. Some studies claim an antiangiogenic effect and others claim a pro-angiogenic one. Thus, the aim of this study was to determine if genistein may exhibit bivalent angiogenic effects. To address this question, genistein angiogenic modulatory effects were examined using an in vitro 3D angiogenesis model using human umbilical vein endothelial cells. In this model, a bivalent effect of genistein was demonstrated on sprouting angiogenesis, with angiogenic stimulation at low concentrations (0.001 – 1 µM) and inhibition at higher ones (25 – 100 µM). Enhancement of the endothelial tube formation correlated with an increase in human umbilical vein endothelial cell metabolic activity and proliferation. Inhibition of angiogenesis correlated with a decreased metabolic activity, proliferation, and migration. Moreover, high concentrations of genistein influenced human umbilical vein endothelial cell morphology. Expression of genes involved in the angiogenic process in response to genistein was measured to study the mechanism of action. Secretome profiling revealed that angiogenic regulators were modulated with genistein treatment. These results suggested a bivalent effect of genistein on human umbilical vein endothelial cell growth and angiogenesis, and further investigations on the benefit of genistein for cancer chemoprevention, cancer treatment, or pro-angiogenic therapies have to be carefully considered.

scholarly journals Alginate-amphotericin B nanocomplexes covered by nanocrystals from bacterial cellulose: physico-chemical characterization and in vitro toxicity

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Victória Soares Soeiro ◽  
Ricardo Silva-Carvalho ◽  
Daniela Martins ◽  
Pier Parpot ◽  
Denise Grotto ◽  
...  
Keyword(s):  
Bacterial Cellulose ◽  
Amphotericin B ◽  
Water Solubility ◽  
Chemical Characterization ◽  
Water Soluble ◽  
In Vitro Toxicity ◽  
Physico Chemical ◽  
Additional Protection ◽  
Anti Fungal

AbstractNanocomplexes systems made up natural poylymers have pharmacotechnical advantages such as increase of water solubility and a decrease of drugs toxicity. Amphotericin B (AmB) is a drug apply as anti-leishmanial and anti-fungal, however it has low water solubility and high toxicity, limiting its therapeutic application. With this in mind, the present study aimed to produce nanocomplexes composed by alginate (Alg), a natural polymer, with AmB covered by nanocrystals from bacterial cellulose (CNC). For this reason, the nanocomplexes were produced utilizing sodium alginate, amphotericin B in a borate buffer (pH 11.0). The CNC was obtained by enzymatic hydrolysis of the bacterial cellulose. To CNC cover the nanocomplexes 1 ml of the nanocomplexes was added into 1 ml of 0.01% CNC suspension. The results showed an ionic adsorption of the CNC into the Alg-AmB nanocomplexes surface. This phenomena was confirmed by an increase in the particle size and PDI decrease. Besides, nanocomplexes samples covered by CNC showed uniformity. The amorphous inclusion of AmB complex into the polysaccharide chain network in both formulations. AmB in the nanocomplexes was in supper-aggregated form and showed good biocompatibility, being significantly less cytotoxic in vitro against kidney cells and significantly less hemolytic compared to the free-drug. The in vitro toxicity results indicated the Alg-AmB nanocomplexes can be considered a non-toxic alternative to improve the AmB therapeutic effect. All process to obtain nanocomplexes and it coat was conduce without organic solvents, can be considered a green process, and allowed to obtain water soluble particles. Furthermore, CNC covering the nanocomplexes brought additional protection to the system can contribut advancement in the pharmaceutical.

scholarly journals Alginate-Amphothericin B Nanocomplexes Covered By Nanocrystals From Bacterial Cellulose: Physico-Chemical Characterization And In Vitro Toxicity

2021 ◽  
Author(s):  
Victória Soares Soeiro ◽  
Ricardo Silva-Carvalho ◽  
Daniela Martins ◽  
Pier Parpot ◽  
Denise Grotto ◽  
...  
Keyword(s):  
Water Solubility ◽  
Chemical Characterization ◽  
Water Soluble ◽  
In Vitro Toxicity ◽  
Small Surface ◽  
Physico Chemical ◽  
Nanostructured Systems ◽  
Additional Protection ◽  
Anti Fungal

Abstract Amphotericin B (AmB) is a drug with anti-leishmanial and anti-fungal ability, but it has low water solubility and permeability, limiting its therapeutic use. Therefore, the incorporation of AmB into nanostructured systems could be profit. Nanostructured systems without surfactants have pharmacotechnical advantages such as amplify the water solubility and decrease the toxicity. For this reason, the present study aimed to produce a nanocomplex of alginate (Alg) with AmB covered by NCC in order to decrease the toxicity of AmB. This straightforward process allowed to obtain water soluble particles Furthermore, the ionic adsorption of the NCC into the Alg-AmB nanocomplex surface was confirmed by an increase in the particle size and a small surface charge decrease. The amorphous inclusion of AmB complex into the polysaccharide chain network in both formulations. AmB in the nanocomplexes was in supper-aggregated form and showed good biocompatibility, being significantly less cytotoxic in vitro against kidney cells and significantly less hemolytic compared to the free-drug. The results indicated the Alg-AmB nanocomplex can be considered a non-toxic alternative to improve the AmB therapeutic effect. Furthermore, NCC coating of the nanocomplexes brought additional protection to the system without compromised the advantages attributed to the developed formulation.

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