13 research outputs found
Additional file 3: of Endothelial properties of third-trimester amniotic fluid stem cells cultured in hypoxia
Adipogenic and osteogenic differentiation of amniotic fluid stem cells from both trimesters. (A) Oil Red O staining (left images) (scale bar = 200 μm) and relative quantification (right graph) (expressed by ratio compared with control cells) (n = 5) performed at day 21 of differentiation. (B) Osteogenic differentiation was attained and calcium salt accumulation was detected by Von Kossa staining (top left images, brown-black deposits) (scale bar = 100 μm); calcium deposits from differentiating cells were also extracted and quantified at different time points (top right graph) (n = 3). Alkaline phosphatase activity was observed by colorimetric substrate modification (bottom left images) on differentiating cells at different time points. Alkaline phosphatase activity was also relatively quantified by measuring the substrate absorbance after reaction (bottom right graph) (expressed by ratio of differentiating over control cells, n = 3). **P < 0.01, ***P < 0.001. Ab antibody, ALP alkaline phosphatase. (JPEG 8213 kb
Additional file 2: of Endothelial properties of third-trimester amniotic fluid stem cells cultured in hypoxia
Cell characterization. (A) SSEA-4 staining for AFS cells in all culture conditions. (B) Percentage of SSEA-4-positive AFS cells in all conditions. AFS amniotic fluid stem, SSEA-4 stage-specific embryonic antigen-4 (JPEG 2556 kb
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps
C1q-Mediated Complement Activation and C3 Opsonization Trigger Recognition of Stealth Poly(2-methyl-2-oxazoline)-Coated Silica Nanoparticles by Human Phagocytes
PolyÂ(2-methyl-2-oxazoline) (PMOXA)
is an alternative promising polymer to polyÂ(ethylene glycol) (PEG)
for design and engineering of macrophage-evading nanoparticles (NPs).
Although PMOXA-engineered NPs have shown comparable pharmacokinetics
and <i>in vivo</i> performance to PEGylated stealth NPs
in the murine model, its interaction with elements of the human innate
immune system has not been studied. From a translational angle, we
studied the interaction of fully characterized PMOXA-coated vinyltriethoxysilane-derived
organically modified silica NPs (PMOXA-coated NPs) of approximately
100 nm in diameter with human complement system, blood leukocytes,
and macrophages and compared their performance with PEGylated and
uncoated NP counterparts. Through detailed immunological and proteomic
profiling, we show that PMOXA-coated NPs extensively trigger complement
activation in human sera exclusively through the classical pathway.
Complement activation is initiated by the sensing molecule C1q, where
C1q binds with high affinity (<i>K</i><sub>d</sub> = 11
± 1 nM) to NP surfaces independent of immunoglobulin binding.
C1q-mediated complement activation accelerates PMOXA opsonization
with the third complement protein (C3) through the amplification loop
of the alternative pathway. This promoted NP recognition by human
blood leukocytes and monocyte-derived macrophages. The macrophage
capture of PMOXA-coated NPs correlates with sera donor variability
in complement activation and opsonization but not with other major
corona proteins, including clusterin and a wide range of apolipoproteins. In contrast to these observations, PMOXA-coated NPs poorly activated the murine complement system and were marginally recognized by mouse macrophages. These studies provide important insights into compatibility of engineered NPs with elements of the human innate immune system for translational steps