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Cell-adhesive RGD peptide-displaying M13 bacteriophage/PLGA nanofiber matrices for growth of fibroblasts.

Shin YC, Lee JH, Jin L, Kim MJ, Oh JW, Kim TW, Han DW - Biomater Res (2014)

Bottom Line: The surface morphology and chemical composition of hybrid nanofiber matrices were characterized by scanning electron microscopy (SEM) and Raman spectroscopy, respectively.Immunofluorescence images and Raman spectra revealed that RGD-M13 phages were homogeneously distributed in entire matrices.These results suggest that RGD-M13 phage/PLGA matrices can be efficiently used as biomimetic scaffolds for tissue engineering applications.

View Article: PubMed Central - PubMed

Affiliation: Department of Cogno-Mechatronics Engineering, Pusan National University, Busan, 609-735 Korea.

ABSTRACT

Background: M13 bacteriophages can be readily fabricated as nanofibers due to non-toxic bacterial virus with a nanofiber-like shape. In the present study, we prepared hybrid nanofiber matrices composed of poly(lactic-co-glycolic acid, PLGA) and M13 bacteriophages which were genetically modified to display the RGD peptide on their surface (RGD-M13 phage).

Results: The surface morphology and chemical composition of hybrid nanofiber matrices were characterized by scanning electron microscopy (SEM) and Raman spectroscopy, respectively. Immunofluorescence staining was conducted to investigate the existence of M13 bacteriophages in RGD-M13 phage/PLGA hybrid nanofibers. In addition, the attachment and proliferation of three different types of fibroblasts on RGD-M13 phage/PLGA nanofiber matrices were evaluated to explore how fibroblasts interact with these matrices. SEM images showed that RGD-M13 phage/PLGA hybrid matrices had the non-woven porous structure, quite similar to that of natural extracellular matrices, having an average fiber diameter of about 190 nm. Immunofluorescence images and Raman spectra revealed that RGD-M13 phages were homogeneously distributed in entire matrices. Moreover, the attachment and proliferation of fibroblasts cultured on RGD-M13 phage/PLGA matrices were significantly enhanced due to enriched RGD moieties on hybrid matrices.

Conclusions: These results suggest that RGD-M13 phage/PLGA matrices can be efficiently used as biomimetic scaffolds for tissue engineering applications.

No MeSH data available.


Related in: MedlinePlus

Surface morphology and immunostaining of electrospun nanofibers. (A, B) SEM images of RGD-M13 phage/PLGA nanofibers. Fluorescence microscopic images of RGD-M13 phage/PLGA nanofibers (C) and pure PLGA nanofibers (D). RGD-M13 phages in RGD-M13 phage/PLGA nanofibers were immunostained with FITC-labeled anti-M13 phage antibody and fluoresced green.
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Fig2: Surface morphology and immunostaining of electrospun nanofibers. (A, B) SEM images of RGD-M13 phage/PLGA nanofibers. Fluorescence microscopic images of RGD-M13 phage/PLGA nanofibers (C) and pure PLGA nanofibers (D). RGD-M13 phages in RGD-M13 phage/PLGA nanofibers were immunostained with FITC-labeled anti-M13 phage antibody and fluoresced green.

Mentions: As shown in Figure 1, RGD-M13 phages and PLGA were blended and then fabricated into hybrid nanofibers thru an electrospinning technique. The artificial scaffolds should be similar to structural property of the natural ECM and support cell growth through 3D microenvironment. As shown in Figure 2A, the morphology of RGD-M13 phage/PLGA nanofibers was not only non-woven porous but also uniform and bead-less as similar to the natural ECM. The RGD-M13 phage/PLGA nanofibers had an average diameter of 190 ± 30 nm (Figure 2B). The RGD-M13 phages of electrospun nanofibers could be very well interaction with cells because the matrices consisted of nanometer scale fibers had a very high surface area-to-volume ratio. Immunostaining fluorescence image showed that genetically engineered RGD-M13 phages were located on the hybrid nanofibers (Figure 2C). Green fluorescence from RGD-M13 phages labeled with FITC was detected along the RGD-M13 phage/PLGA nanofibers. On the contrary, any fluorescence was not exhibited from pure PLGA nanofibers (Figure 2D).Figure 2


Cell-adhesive RGD peptide-displaying M13 bacteriophage/PLGA nanofiber matrices for growth of fibroblasts.

Shin YC, Lee JH, Jin L, Kim MJ, Oh JW, Kim TW, Han DW - Biomater Res (2014)

Surface morphology and immunostaining of electrospun nanofibers. (A, B) SEM images of RGD-M13 phage/PLGA nanofibers. Fluorescence microscopic images of RGD-M13 phage/PLGA nanofibers (C) and pure PLGA nanofibers (D). RGD-M13 phages in RGD-M13 phage/PLGA nanofibers were immunostained with FITC-labeled anti-M13 phage antibody and fluoresced green.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4552277&req=5

Fig2: Surface morphology and immunostaining of electrospun nanofibers. (A, B) SEM images of RGD-M13 phage/PLGA nanofibers. Fluorescence microscopic images of RGD-M13 phage/PLGA nanofibers (C) and pure PLGA nanofibers (D). RGD-M13 phages in RGD-M13 phage/PLGA nanofibers were immunostained with FITC-labeled anti-M13 phage antibody and fluoresced green.
Mentions: As shown in Figure 1, RGD-M13 phages and PLGA were blended and then fabricated into hybrid nanofibers thru an electrospinning technique. The artificial scaffolds should be similar to structural property of the natural ECM and support cell growth through 3D microenvironment. As shown in Figure 2A, the morphology of RGD-M13 phage/PLGA nanofibers was not only non-woven porous but also uniform and bead-less as similar to the natural ECM. The RGD-M13 phage/PLGA nanofibers had an average diameter of 190 ± 30 nm (Figure 2B). The RGD-M13 phages of electrospun nanofibers could be very well interaction with cells because the matrices consisted of nanometer scale fibers had a very high surface area-to-volume ratio. Immunostaining fluorescence image showed that genetically engineered RGD-M13 phages were located on the hybrid nanofibers (Figure 2C). Green fluorescence from RGD-M13 phages labeled with FITC was detected along the RGD-M13 phage/PLGA nanofibers. On the contrary, any fluorescence was not exhibited from pure PLGA nanofibers (Figure 2D).Figure 2

Bottom Line: The surface morphology and chemical composition of hybrid nanofiber matrices were characterized by scanning electron microscopy (SEM) and Raman spectroscopy, respectively.Immunofluorescence images and Raman spectra revealed that RGD-M13 phages were homogeneously distributed in entire matrices.These results suggest that RGD-M13 phage/PLGA matrices can be efficiently used as biomimetic scaffolds for tissue engineering applications.

View Article: PubMed Central - PubMed

Affiliation: Department of Cogno-Mechatronics Engineering, Pusan National University, Busan, 609-735 Korea.

ABSTRACT

Background: M13 bacteriophages can be readily fabricated as nanofibers due to non-toxic bacterial virus with a nanofiber-like shape. In the present study, we prepared hybrid nanofiber matrices composed of poly(lactic-co-glycolic acid, PLGA) and M13 bacteriophages which were genetically modified to display the RGD peptide on their surface (RGD-M13 phage).

Results: The surface morphology and chemical composition of hybrid nanofiber matrices were characterized by scanning electron microscopy (SEM) and Raman spectroscopy, respectively. Immunofluorescence staining was conducted to investigate the existence of M13 bacteriophages in RGD-M13 phage/PLGA hybrid nanofibers. In addition, the attachment and proliferation of three different types of fibroblasts on RGD-M13 phage/PLGA nanofiber matrices were evaluated to explore how fibroblasts interact with these matrices. SEM images showed that RGD-M13 phage/PLGA hybrid matrices had the non-woven porous structure, quite similar to that of natural extracellular matrices, having an average fiber diameter of about 190 nm. Immunofluorescence images and Raman spectra revealed that RGD-M13 phages were homogeneously distributed in entire matrices. Moreover, the attachment and proliferation of fibroblasts cultured on RGD-M13 phage/PLGA matrices were significantly enhanced due to enriched RGD moieties on hybrid matrices.

Conclusions: These results suggest that RGD-M13 phage/PLGA matrices can be efficiently used as biomimetic scaffolds for tissue engineering applications.

No MeSH data available.


Related in: MedlinePlus