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Biomimetic Hybrid Nanofiber Sheets Composed of RGD Peptide-Decorated PLGA as Cell-Adhesive Substrates.

Shin YC, Lee JH, Kim MJ, Park JH, Kim SE, Kim JS, Oh JW, Han DW - J Funct Biomater (2015)

Bottom Line: RGD peptide-decorated PLGA (RGD-PLGA) nanofiber sheets were characterized by scanning electron microscopy, immunofluorescence staining, contact angle measurement and differential scanning calorimetry.Our results showed that the hybrid nanofiber sheets have a three-dimensional porous structure comparable to the native ECM.These results suggest that biomimetic RGD-PLGA nanofiber sheets can be promising cell-adhesive substrates for application as tissue engineering scaffolds.

View Article: PubMed Central - PubMed

Affiliation: Department of Optics and Mechatronics Engineering, BK21+ Nano-Integrated Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University, Busandaehak-ro 63beon-gil, Geumjeong-gu, Busan 609-735, Korea. choel15@naver.com.

ABSTRACT
In biomedical applications, there is a need for tissue engineering scaffolds to promote and control cellular behaviors, including adhesion, proliferation and differentiation. In particular, the initial adhesion of cells has a great influence on those cellular behaviors. In this study, we concentrate on developing cell-adhesive substrates applicable for tissue engineering scaffolds. The hybrid nanofiber sheets were prepared by electrospinning poly(lactic-co-glycolic acid) (PLGA) and M13 phage, which was genetically modified to enhance cell adhesion thru expressing RGD peptides on their surface. The RGD peptide is a specific motif of extracellular matrix (ECM) for integrin receptors of cells. RGD peptide-decorated PLGA (RGD-PLGA) nanofiber sheets were characterized by scanning electron microscopy, immunofluorescence staining, contact angle measurement and differential scanning calorimetry. In addition, the initial adhesion and proliferation of four different types of mammalian cells were determined in order to evaluate the potential of RGD-PLGA nanofiber sheets as cell-adhesive substrates. Our results showed that the hybrid nanofiber sheets have a three-dimensional porous structure comparable to the native ECM. Furthermore, the initial adhesion and proliferation of cells were significantly enhanced on RGD-PLGA sheets. These results suggest that biomimetic RGD-PLGA nanofiber sheets can be promising cell-adhesive substrates for application as tissue engineering scaffolds.

No MeSH data available.


Related in: MedlinePlus

(A) Representative SEM image of RGD-PLGA nanofiber sheets; (B) representative immunofluorescence images of the pure PLGA and RGD-PLGA nanofiber sheets. RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the fluorescein isothiocyanate (FITC)-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results.
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jfb-06-00367-f002: (A) Representative SEM image of RGD-PLGA nanofiber sheets; (B) representative immunofluorescence images of the pure PLGA and RGD-PLGA nanofiber sheets. RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the fluorescein isothiocyanate (FITC)-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results.

Mentions: RGD-PLGA nanofiber sheets were fabricated by electrospinning of PLGA and RGD-M13 phage blend solutions (Figure 1). The optimal mixing ratio between RGD-M13 phage and PLGA solution was 1:3. The surface morphology of RGD-PLGA nanofiber sheets was observed by SEM, as shown in Figure 2A. The SEM image demonstrated that the RGD-PLGA nanofiber sheet has a three-dimensional porous architecture with interconnected pores homologous to the ECM. In addition, the RGD-PLGA nanofibers have continuous, smooth and beadless morphologies with an average diameter of 370 ± 190 nm. This indicates that the electrospinning parameters, including the mixing ratio, voltage, distance between the tip of the needle and the collector and the flow rate of a blend solution were optimum for the fabrication of RGD-PLGA nanofibers. These nanometer-scale diameters of RGD-PLGA hybrid fibers allow achieving a high surface area-to-volume ratio [28]. Therefore, the RGD-PLGA nanofiber sheets can effectively interact with cells. The distribution of decorated RGD peptides on the nanofiber sheets was examined by immunofluorescence staining for RGD-M13 phages (Figure 2B). The RGD-PLGA nanofiber sheet showed green fluorescence of RGD-M13 phages throughout the sheets, whereas the pure PLGA nanofiber sheet did not show any detectable fluorescence. Therefore, it was revealed that the RGD-PLGA nanofiber sheets were successfully prepared, and the RGD peptides were abundantly decorated on the sheets.


Biomimetic Hybrid Nanofiber Sheets Composed of RGD Peptide-Decorated PLGA as Cell-Adhesive Substrates.

Shin YC, Lee JH, Kim MJ, Park JH, Kim SE, Kim JS, Oh JW, Han DW - J Funct Biomater (2015)

(A) Representative SEM image of RGD-PLGA nanofiber sheets; (B) representative immunofluorescence images of the pure PLGA and RGD-PLGA nanofiber sheets. RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the fluorescein isothiocyanate (FITC)-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4493517&req=5

jfb-06-00367-f002: (A) Representative SEM image of RGD-PLGA nanofiber sheets; (B) representative immunofluorescence images of the pure PLGA and RGD-PLGA nanofiber sheets. RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the fluorescein isothiocyanate (FITC)-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results.
Mentions: RGD-PLGA nanofiber sheets were fabricated by electrospinning of PLGA and RGD-M13 phage blend solutions (Figure 1). The optimal mixing ratio between RGD-M13 phage and PLGA solution was 1:3. The surface morphology of RGD-PLGA nanofiber sheets was observed by SEM, as shown in Figure 2A. The SEM image demonstrated that the RGD-PLGA nanofiber sheet has a three-dimensional porous architecture with interconnected pores homologous to the ECM. In addition, the RGD-PLGA nanofibers have continuous, smooth and beadless morphologies with an average diameter of 370 ± 190 nm. This indicates that the electrospinning parameters, including the mixing ratio, voltage, distance between the tip of the needle and the collector and the flow rate of a blend solution were optimum for the fabrication of RGD-PLGA nanofibers. These nanometer-scale diameters of RGD-PLGA hybrid fibers allow achieving a high surface area-to-volume ratio [28]. Therefore, the RGD-PLGA nanofiber sheets can effectively interact with cells. The distribution of decorated RGD peptides on the nanofiber sheets was examined by immunofluorescence staining for RGD-M13 phages (Figure 2B). The RGD-PLGA nanofiber sheet showed green fluorescence of RGD-M13 phages throughout the sheets, whereas the pure PLGA nanofiber sheet did not show any detectable fluorescence. Therefore, it was revealed that the RGD-PLGA nanofiber sheets were successfully prepared, and the RGD peptides were abundantly decorated on the sheets.

Bottom Line: RGD peptide-decorated PLGA (RGD-PLGA) nanofiber sheets were characterized by scanning electron microscopy, immunofluorescence staining, contact angle measurement and differential scanning calorimetry.Our results showed that the hybrid nanofiber sheets have a three-dimensional porous structure comparable to the native ECM.These results suggest that biomimetic RGD-PLGA nanofiber sheets can be promising cell-adhesive substrates for application as tissue engineering scaffolds.

View Article: PubMed Central - PubMed

Affiliation: Department of Optics and Mechatronics Engineering, BK21+ Nano-Integrated Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University, Busandaehak-ro 63beon-gil, Geumjeong-gu, Busan 609-735, Korea. choel15@naver.com.

ABSTRACT
In biomedical applications, there is a need for tissue engineering scaffolds to promote and control cellular behaviors, including adhesion, proliferation and differentiation. In particular, the initial adhesion of cells has a great influence on those cellular behaviors. In this study, we concentrate on developing cell-adhesive substrates applicable for tissue engineering scaffolds. The hybrid nanofiber sheets were prepared by electrospinning poly(lactic-co-glycolic acid) (PLGA) and M13 phage, which was genetically modified to enhance cell adhesion thru expressing RGD peptides on their surface. The RGD peptide is a specific motif of extracellular matrix (ECM) for integrin receptors of cells. RGD peptide-decorated PLGA (RGD-PLGA) nanofiber sheets were characterized by scanning electron microscopy, immunofluorescence staining, contact angle measurement and differential scanning calorimetry. In addition, the initial adhesion and proliferation of four different types of mammalian cells were determined in order to evaluate the potential of RGD-PLGA nanofiber sheets as cell-adhesive substrates. Our results showed that the hybrid nanofiber sheets have a three-dimensional porous structure comparable to the native ECM. Furthermore, the initial adhesion and proliferation of cells were significantly enhanced on RGD-PLGA sheets. These results suggest that biomimetic RGD-PLGA nanofiber sheets can be promising cell-adhesive substrates for application as tissue engineering scaffolds.

No MeSH data available.


Related in: MedlinePlus