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Evaluation of silica nanoparticle binding to major human blood proteins.

Hata K, Higashisaka K, Nagano K, Mukai Y, Kamada H, Tsunoda S, Yoshioka Y, Tsutsumi Y - Nanoscale Res Lett (2014)

Bottom Line: Nanomaterials are used for various biomedical applications because they are often more effective than conventional materials.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the amount of albumin, transferrin, and IgG binding to the silica particles increased as the particle size decreased under conditions where the silica particle mass remained the same.These results suggest that the characteristics of nanomaterials are important for binding with human blood proteins; this information may contribute to the development of safe and effective nanomaterials.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka, 565-0871, Japan, k-hata@towayakuhin.co.jp.

ABSTRACT
Nanomaterials are used for various biomedical applications because they are often more effective than conventional materials. Recently, however, it has become clear that the protein corona that forms on the surface of nanomaterials when they make contact with biological fluids, such as blood, influences the pharmacokinetics and biological responses induced by the nanomaterials. Therefore, when evaluating nanomaterial safety and efficacy, it is important to analyze the interaction between nanomaterials and proteins in biological fluids and to evaluate the effects of the protein corona. Here, we evaluated the interaction of silica nanoparticles, a commonly used nanomaterial, with the human blood proteins albumin, transferrin, fibrinogen, and IgG. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the amount of albumin, transferrin, and IgG binding to the silica particles increased as the particle size decreased under conditions where the silica particle mass remained the same. However, under conditions in which the specific surface area remained constant, there were no differences in the binding of human plasma proteins to the silica particles tested, suggesting that the binding of silica particles with human plasma proteins is dependent on the specific surface area of the silica particles. Furthermore, the amount of albumin, transferrin, and IgG binding to silica nanoparticles with a diameter of 70 nm (nSP70) and a functional amino group was lower than that with unmodified nSP70, although there was no difference in the binding between nSP70 with the surface modification of a carboxyl functional group and nSP70. These results suggest that the characteristics of nanomaterials are important for binding with human blood proteins; this information may contribute to the development of safe and effective nanomaterials.

No MeSH data available.


Effects of silica particle size on binding to human plasma proteins. Each protein solution was mixed with SP70, nSP100, nSP300, or mSP1000 (25 mg/mL). After centrifugation, each sample was separated by SDS-PAGE. The gel was stained by CBB staining, and the protein bands of albumin (A), transferrin (B), IgG (C), and fibrinogen (D) were quantified with the ImageJ software. The protein content of each band was estimated from its optical density compared with the optical density of each standard solution. Data are presented as the mean ± SD; n = 4; **P < 0.01 vs. nSP70-treated group; *P < 0.05 vs. nSP70-treated group; N.D., not detected.
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Fig1: Effects of silica particle size on binding to human plasma proteins. Each protein solution was mixed with SP70, nSP100, nSP300, or mSP1000 (25 mg/mL). After centrifugation, each sample was separated by SDS-PAGE. The gel was stained by CBB staining, and the protein bands of albumin (A), transferrin (B), IgG (C), and fibrinogen (D) were quantified with the ImageJ software. The protein content of each band was estimated from its optical density compared with the optical density of each standard solution. Data are presented as the mean ± SD; n = 4; **P < 0.01 vs. nSP70-treated group; *P < 0.05 vs. nSP70-treated group; N.D., not detected.

Mentions: Next, we evaluated whether the differences in the size of the silica particles affected their binding to albumin, transferrin, fibrinogen, and IgG. We selected these proteins because these have high concentrations in human blood [13] and because the frequency of contact in the body is considered to be high. In addition, it was reported that these proteins were identified in protein corona on silica nanoparticles with human plasma [14, 15]. SDS-PAGE analysis showed that there was no interaction between the silica particles and AGP (data not shown). For albumin (Figure 1A) and IgG (Figure 1C), the binding was dependent on the diameter of the silica particles. For transferrin, we found that nSP300 and mSP1000 did not bind (Figure 1B). In addition, the degree of binding of mSP1000 with fibrinogen was significantly lower than that of the other particles (Figure 1D). These results indicate that the binding of silica particles to albumin and IgG appears to increase as the particle size decreases. The degree of nSP70 binding was greatest with fibrinogen and smallest with albumin among these four proteins tested.Figure 1


Evaluation of silica nanoparticle binding to major human blood proteins.

Hata K, Higashisaka K, Nagano K, Mukai Y, Kamada H, Tsunoda S, Yoshioka Y, Tsutsumi Y - Nanoscale Res Lett (2014)

Effects of silica particle size on binding to human plasma proteins. Each protein solution was mixed with SP70, nSP100, nSP300, or mSP1000 (25 mg/mL). After centrifugation, each sample was separated by SDS-PAGE. The gel was stained by CBB staining, and the protein bands of albumin (A), transferrin (B), IgG (C), and fibrinogen (D) were quantified with the ImageJ software. The protein content of each band was estimated from its optical density compared with the optical density of each standard solution. Data are presented as the mean ± SD; n = 4; **P < 0.01 vs. nSP70-treated group; *P < 0.05 vs. nSP70-treated group; N.D., not detected.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493834&req=5

Fig1: Effects of silica particle size on binding to human plasma proteins. Each protein solution was mixed with SP70, nSP100, nSP300, or mSP1000 (25 mg/mL). After centrifugation, each sample was separated by SDS-PAGE. The gel was stained by CBB staining, and the protein bands of albumin (A), transferrin (B), IgG (C), and fibrinogen (D) were quantified with the ImageJ software. The protein content of each band was estimated from its optical density compared with the optical density of each standard solution. Data are presented as the mean ± SD; n = 4; **P < 0.01 vs. nSP70-treated group; *P < 0.05 vs. nSP70-treated group; N.D., not detected.
Mentions: Next, we evaluated whether the differences in the size of the silica particles affected their binding to albumin, transferrin, fibrinogen, and IgG. We selected these proteins because these have high concentrations in human blood [13] and because the frequency of contact in the body is considered to be high. In addition, it was reported that these proteins were identified in protein corona on silica nanoparticles with human plasma [14, 15]. SDS-PAGE analysis showed that there was no interaction between the silica particles and AGP (data not shown). For albumin (Figure 1A) and IgG (Figure 1C), the binding was dependent on the diameter of the silica particles. For transferrin, we found that nSP300 and mSP1000 did not bind (Figure 1B). In addition, the degree of binding of mSP1000 with fibrinogen was significantly lower than that of the other particles (Figure 1D). These results indicate that the binding of silica particles to albumin and IgG appears to increase as the particle size decreases. The degree of nSP70 binding was greatest with fibrinogen and smallest with albumin among these four proteins tested.Figure 1

Bottom Line: Nanomaterials are used for various biomedical applications because they are often more effective than conventional materials.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the amount of albumin, transferrin, and IgG binding to the silica particles increased as the particle size decreased under conditions where the silica particle mass remained the same.These results suggest that the characteristics of nanomaterials are important for binding with human blood proteins; this information may contribute to the development of safe and effective nanomaterials.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka, 565-0871, Japan, k-hata@towayakuhin.co.jp.

ABSTRACT
Nanomaterials are used for various biomedical applications because they are often more effective than conventional materials. Recently, however, it has become clear that the protein corona that forms on the surface of nanomaterials when they make contact with biological fluids, such as blood, influences the pharmacokinetics and biological responses induced by the nanomaterials. Therefore, when evaluating nanomaterial safety and efficacy, it is important to analyze the interaction between nanomaterials and proteins in biological fluids and to evaluate the effects of the protein corona. Here, we evaluated the interaction of silica nanoparticles, a commonly used nanomaterial, with the human blood proteins albumin, transferrin, fibrinogen, and IgG. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the amount of albumin, transferrin, and IgG binding to the silica particles increased as the particle size decreased under conditions where the silica particle mass remained the same. However, under conditions in which the specific surface area remained constant, there were no differences in the binding of human plasma proteins to the silica particles tested, suggesting that the binding of silica particles with human plasma proteins is dependent on the specific surface area of the silica particles. Furthermore, the amount of albumin, transferrin, and IgG binding to silica nanoparticles with a diameter of 70 nm (nSP70) and a functional amino group was lower than that with unmodified nSP70, although there was no difference in the binding between nSP70 with the surface modification of a carboxyl functional group and nSP70. These results suggest that the characteristics of nanomaterials are important for binding with human blood proteins; this information may contribute to the development of safe and effective nanomaterials.

No MeSH data available.