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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: 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.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

Schematic diagrams of RGD peptide-displaying M13 bacteriophage and electrospinning process for fabrication of hybrid nanofibers. (A) DNA sequence of wild-type M13 bacteriophages. (B) DNA sequence of RGD peptide-displaying M13 bacteriophages (RGD-M13 phages). (C) Structure of genetically engineered RGD-M13 phage. (D) Fabrication of RGD-M13 phage/PLGA nanofibers by electrospinning.
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Fig1: Schematic diagrams of RGD peptide-displaying M13 bacteriophage and electrospinning process for fabrication of hybrid nanofibers. (A) DNA sequence of wild-type M13 bacteriophages. (B) DNA sequence of RGD peptide-displaying M13 bacteriophages (RGD-M13 phages). (C) Structure of genetically engineered RGD-M13 phage. (D) Fabrication of RGD-M13 phage/PLGA nanofibers by electrospinning.

Mentions: Genetic engineering of M13 bacteriophage was conducted according to the method as previously described [20]. In brief, to display a desired peptide on major coat protein of M13 bacteriophage surface, an inverse PCR cloning method was carried out as described elsewhere [24, 27, 28]. As shown in Figure 1A-C, we built RGD-M13 phages through genetic modification of wild-type M13 bacteriophages. PLGA resins (PLA/PGA = 75/25, MW = 70–110 kDa) were purchased from Sigma-Aldrich (St Louis, MO). RGD-M13 phage/PLGA nanofiber matrices were fabricated by an electrospinning process (Figure 1D) as described elsewhere [29]. Random oriented RGD-M13 phage/PLGA nanofibers were collected on a steel rotating wheel covered with an aluminum foil. Then, electrospun nanofiber matrices were dried overnight under vacuum at room temperature in order to remove any residual solvent.Figure 1


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)

Schematic diagrams of RGD peptide-displaying M13 bacteriophage and electrospinning process for fabrication of hybrid nanofibers. (A) DNA sequence of wild-type M13 bacteriophages. (B) DNA sequence of RGD peptide-displaying M13 bacteriophages (RGD-M13 phages). (C) Structure of genetically engineered RGD-M13 phage. (D) Fabrication of RGD-M13 phage/PLGA nanofibers by electrospinning.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig1: Schematic diagrams of RGD peptide-displaying M13 bacteriophage and electrospinning process for fabrication of hybrid nanofibers. (A) DNA sequence of wild-type M13 bacteriophages. (B) DNA sequence of RGD peptide-displaying M13 bacteriophages (RGD-M13 phages). (C) Structure of genetically engineered RGD-M13 phage. (D) Fabrication of RGD-M13 phage/PLGA nanofibers by electrospinning.
Mentions: Genetic engineering of M13 bacteriophage was conducted according to the method as previously described [20]. In brief, to display a desired peptide on major coat protein of M13 bacteriophage surface, an inverse PCR cloning method was carried out as described elsewhere [24, 27, 28]. As shown in Figure 1A-C, we built RGD-M13 phages through genetic modification of wild-type M13 bacteriophages. PLGA resins (PLA/PGA = 75/25, MW = 70–110 kDa) were purchased from Sigma-Aldrich (St Louis, MO). RGD-M13 phage/PLGA nanofiber matrices were fabricated by an electrospinning process (Figure 1D) as described elsewhere [29]. Random oriented RGD-M13 phage/PLGA nanofibers were collected on a steel rotating wheel covered with an aluminum foil. Then, electrospun nanofiber matrices were dried overnight under vacuum at room temperature in order to remove any residual solvent.Figure 1

Bottom Line: 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.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