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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

Two-photon excitation fluorescence images of HASMCs on the pure PLGA and RGD-PLGA nanofiber sheets. F-actins of HASMC cytoskeletons were stained with tetramethylrhodamine isothiocyanate (TRITC)-labelled phalloidin (red); cell nuclei were counter stained with 4',6-diamidino-2-phenylindole (DAPI, blue); and RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the FITC-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results. (A) Low-magnification and (B) higher-magnification images of HASMCs on the RGD-PLGA nanofiber sheets; (C) low-magnification and (D) higher-magnification images of HASMCs on the pure PLGA nanofiber sheets.
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jfb-06-00367-f006: Two-photon excitation fluorescence images of HASMCs on the pure PLGA and RGD-PLGA nanofiber sheets. F-actins of HASMC cytoskeletons were stained with tetramethylrhodamine isothiocyanate (TRITC)-labelled phalloidin (red); cell nuclei were counter stained with 4',6-diamidino-2-phenylindole (DAPI, blue); and RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the FITC-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results. (A) Low-magnification and (B) higher-magnification images of HASMCs on the RGD-PLGA nanofiber sheets; (C) low-magnification and (D) higher-magnification images of HASMCs on the pure PLGA nanofiber sheets.

Mentions: To confirm our findings of the promoted initial cell adhesions and proliferations, the adherent morphologies of HASMCs, cultured on the pure PLGA and RGD-PLGA nanofiber sheets for three days, were imaged with the two-photon excitation microscope. Figure 6 shows the obvious difference in HASMC morphologies between the pure PLGA nanofiber sheets and the RGD-PLGA nanofiber sheets. The HASMCs on the RGD-PLGA nanofiber sheets were successfully grown and maintained their well-adhered morphology (Figure 6A). As shown in the higher magnification image, they showed a well-organized F-actin network (Figure 6B). In addition, the RGD-PLGA nanofiber sheets exhibited green fluorescence throughout the sheet due to the decorated RGD-M13 phage. Contrary to this, the HASMCs on the pure PLGA nanofiber sheets were not fully grown and showed an irregular morphology with poorly-developed F-actins owing to the hydrophobic surface property of the sheets (Figure 6C,D). Therefore, it is suggested that the RGD-PLGA nanofiber sheets can specifically facilitate cellular behaviors, including cell adhesion and proliferation, due to the synergistic effects of an increase in the hydrophilic surface property and the RGD peptides.


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)

Two-photon excitation fluorescence images of HASMCs on the pure PLGA and RGD-PLGA nanofiber sheets. F-actins of HASMC cytoskeletons were stained with tetramethylrhodamine isothiocyanate (TRITC)-labelled phalloidin (red); cell nuclei were counter stained with 4',6-diamidino-2-phenylindole (DAPI, blue); and RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the FITC-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results. (A) Low-magnification and (B) higher-magnification images of HASMCs on the RGD-PLGA nanofiber sheets; (C) low-magnification and (D) higher-magnification images of HASMCs on the pure PLGA nanofiber sheets.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4493517&req=5

jfb-06-00367-f006: Two-photon excitation fluorescence images of HASMCs on the pure PLGA and RGD-PLGA nanofiber sheets. F-actins of HASMC cytoskeletons were stained with tetramethylrhodamine isothiocyanate (TRITC)-labelled phalloidin (red); cell nuclei were counter stained with 4',6-diamidino-2-phenylindole (DAPI, blue); and RGD-M13 phages in the RGD-PLGA nanofiber sheets were immunostained with the FITC-labelled anti-M13 phage antibody (green). All images shown in this figure are representative of six independent experiments with similar results. (A) Low-magnification and (B) higher-magnification images of HASMCs on the RGD-PLGA nanofiber sheets; (C) low-magnification and (D) higher-magnification images of HASMCs on the pure PLGA nanofiber sheets.
Mentions: To confirm our findings of the promoted initial cell adhesions and proliferations, the adherent morphologies of HASMCs, cultured on the pure PLGA and RGD-PLGA nanofiber sheets for three days, were imaged with the two-photon excitation microscope. Figure 6 shows the obvious difference in HASMC morphologies between the pure PLGA nanofiber sheets and the RGD-PLGA nanofiber sheets. The HASMCs on the RGD-PLGA nanofiber sheets were successfully grown and maintained their well-adhered morphology (Figure 6A). As shown in the higher magnification image, they showed a well-organized F-actin network (Figure 6B). In addition, the RGD-PLGA nanofiber sheets exhibited green fluorescence throughout the sheet due to the decorated RGD-M13 phage. Contrary to this, the HASMCs on the pure PLGA nanofiber sheets were not fully grown and showed an irregular morphology with poorly-developed F-actins owing to the hydrophobic surface property of the sheets (Figure 6C,D). Therefore, it is suggested that the RGD-PLGA nanofiber sheets can specifically facilitate cellular behaviors, including cell adhesion and proliferation, due to the synergistic effects of an increase in the hydrophilic surface property and the RGD peptides.

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