Fluorescence labeling to study platelet and leucocyte deposition onto vascular grafts in vitro

Toes, GJ; Dungen, van den Johannes; Haan, J. de; Hermens, R. A. E. C.; Oeveren, van Willem

doi:10.1016/s0142-9612(99)00100-3

Cited by 18 publications

(9 citation statements)

References 12 publications

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“…At the end of the study (Figure 4A), ePTFE tubings were fixed and assessed for platelet aggregation by scanning electron microscopy (Figure 4B) and confocal microscopy (Figure 4D). Platelets and leucocytes are known to be the key responders in small diameter ePTFE grafts [31]–[33]. Assessing the deposition of platelets is known to be important in evaluation of synthetic grafts.…”

Section: Resultsmentioning

confidence: 99%

Human Blood Vessel–Derived Endothelial Progenitors for Endothelialization of Small Diameter Vascular Prosthesis

et al. 2009

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BackgroundCoronary bypass graft failure as a result of acute thrombosis and intimal hyperplasia has been the major challenge in surgical procedures involving small-diameter vascular prosthesis. Coating synthetic grafts with patients' own endothelial cells has been suggested to improve the patency rate and overall success of bypass surgeries.Methodology/Principal FindingsWe isolated endothelial progenitor cells (EPCs) from leftover pieces of human saphenous vein/mammary artery. We demonstrate that EPCs can be expanded to generate millions of cells under low-density culture conditions. Exposure to high-density conditions induces differentiation to endothelial cell phenotype. EPC–derived endothelial cells show expression of CD144high, CD31, and vWF. We then assessed the ability of differentiated endothelial cells to adhere and grow on small diameter expanded polytetrafluoroethylene (ePTFE) tubings. Since ePTFE tubings are highly hydrophobic, we optimized protocols to introduce hydrophilic groups on luminal surface of ePTFE tubings. We demonstrate here a stepwise protocol that involves introduction of hydrophilic moieties and coating with defined ECM components that support adhesion of endothelial cells, but not of blood platelets.Conclusion/SignificanceOur data confirms that endothelial progenitors obtained from adult human blood vessels can be expanded in vitro under xenoprotein-free conditions, for potential use in endothelialization of small diameter ePTFE grafts. These endothelialized grafts may represent a promising treatment strategy for improving the clinical outcome of small-caliber vascular grafts in cardiac bypass surgeries.

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Section: Resultsmentioning

confidence: 99%

Human Blood Vessel–Derived Endothelial Progenitors for Endothelialization of Small Diameter Vascular Prosthesis

et al. 2009

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“…Nanoparticles internalisation has been followed by confocal microscopy analysis, after 1 h incubation at 37 buffer ( Fig. 3(A)).…”

Section: Evolution Of Nanoparticles Load With Cell Growthmentioning

confidence: 99%

Multi-Luminescent Hybrid Gadolinium Oxide Nanoparticles as Potential Cell Labeling

Fizet¹,

Rivière²,

Bridot³

et al. 2009

J. Nanosci. Nanotech.

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This manuscript analyses the use of newly developed hybrid gadolinium oxide nanoparticles as cell-labeling tracers. The nanoparticles are core-shell particles composed of a core of gadolinium oxide of [2-4] nm and a protecting shell of polysiloxane [1-3 nm] where different organic dyes (fluoresceine isothiocyanate (FITC) or rhodamine B isothiocyanate (RBITC)) are embedded. They are functionalized with poly(ethylene glycol)bis(carboxymethyl) to ensure their colloidal stability in biological buffers. These particles are potential multi-labeling tracers (magnetic and optical). In this paper, we show by optical imaging that they can be efficiently internalized in cells without cell alteration. The in-vitro uptake of the nanoparticles was followed in two cell lines (human fibroblasts and a human adenocarnima cell lines MCF7 cells). Nanoparticles distribution within cells was analysed by confocal analysis, and gadolinium concentration within cells was quantified by mass spectrometry (ICP-MS analysis). Nanoparticles uptake is found to be fast and efficient for both cell lines, with fluorescent labeling visible after 10 min of incubation whatever the nature of the fluorophore. The fluorescent intensity is mainly found as concentrated dots in the perinuclear region of the cells and decreases with the number of days in culture, but is still easily detectable after 3 days in culture. No significant effect on cell growth was detected. Finally, we show in this study the protective effect of the polysiloxane layer: encapsulation of RBITC within the polysiloxane shell, leads to a better photostability of this low cost dye than Cy3 and even reach a level comparable to Alexa 595. With their high photostability and long-lasting contrast properties, these hybrid luminescent nanoparticles appears thus as a versatile solution to assess multiple cell fate both in in-vitro cell model as well as in-vivo.

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“…Polyethylene terphthalate Mechanical heart valve 3, 4, 5, 6, 10, 11 Toes (1999), Bonchek et al (1969), Radomski et al (1987), Marois knitted/woven et al (1999), Marois et al (1996), Urayama et al (1996), Granström et al (1986) Biodegradable Polylactic acid Biodegradable pericardial 3, 4, 5, 6, 10, 11 Nguyen et al (2003), (Tamai et al (2000), Kohn et al (2004), replacement Cutright and Hunsuck (1971) Table 3 summarizes the types of biocompatibility issues that might be a consideration in each category of biomaterials described below. These considerations are general and are influenced by the nature of the material (e.g., biostable vs. biodegradable) and application (e.g., soft-tissue, blood, or hard-tissue applications).…”

Section: Synthetic Textilementioning

confidence: 99%

Biocompatibility: Meeting a Key Functional Requirement of Next-Generation Medical Devices

Helmus¹,

Gibbons²,

Cebon

2008

Toxicol Pathol

133

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The array of polymeric, biologic, metallic, and ceramic biomaterials will be reviewed with respect to their biocompatibility, which has traditionally been viewed as a requirement to develop a safe medical device. With the emergence of combination products, a paradigm shift is occurring that now requires biocompatibility to be designed into the device. In fact, next-generation medical devices will require enhanced biocompatibility by using, for example, pharmacological agents, bioactive coatings, nano-textures, or hybrid systems containing cells that control biologic interactions to have desirable biologic outcomes. The concept of biocompatibility is moving from a "do no harm" mission (i.e., nontoxic, nonantigenic, nonmutagenic, etc.) to one of doing "good," that is, encouraging positive healing responses. These new devices will promote the formation of normal healthy tissue as well as the integration of the device into adjacent tissue. In some contexts, biocompatibility can become a disruptive technology that can change therapeutic paradigms (e.g., drug-coated stents). New database tools to access biocompatibility data of the materials of construction in existing medical devices will facilitate the use of existing and new biomaterials for new medical device designs.

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Fluorescence labeling to study platelet and leucocyte deposition onto vascular grafts in vitro

Cited by 18 publications

References 12 publications

Human Blood Vessel–Derived Endothelial Progenitors for Endothelialization of Small Diameter Vascular Prosthesis

Human Blood Vessel–Derived Endothelial Progenitors for Endothelialization of Small Diameter Vascular Prosthesis

Multi-Luminescent Hybrid Gadolinium Oxide Nanoparticles as Potential Cell Labeling

Biocompatibility: Meeting a Key Functional Requirement of Next-Generation Medical Devices

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