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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

Cell attachment of L-929 cells, HDFs and HT-1080 cells on RGD-M13 phage/PLGA nanofiber matrices. Cell attachment was determined by a CCK-8 assay. Data are expressed as mean ± SD based on at least duplicate observations from three independent experiments.
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Fig4: Cell attachment of L-929 cells, HDFs and HT-1080 cells on RGD-M13 phage/PLGA nanofiber matrices. Cell attachment was determined by a CCK-8 assay. Data are expressed as mean ± SD based on at least duplicate observations from three independent experiments.

Mentions: We examined cellular behaviors of fibroblasts on the RGD-M13 phage/PLGA nanofiber matrices. In this study, three types of fibroblast were used: murine fibroblast cell lines (L-929 cells), primary human dermal fibroblasts (HDFs), and human fibrosarcoma cell lines (HT-1080 cells). The cell viability was measured to assess a cell attachment when cells had been seed on the matrices for 6 hours. As shown in Figure 4, fibroblasts showed the highest adhesion where they were cultured on RGD-M13 phage/PLGA matrices regardless of cell types. As mentioned above, RGD peptides contained the matrices have contributed to the enhancement of cell adhesion. In contrast, cell adhesion on pure PLGA matrices was decreased because the PLGA matrices did not contain any materials which could improve cell adhesion. In addition, we evaluated the proliferation of fibroblasts on pure PLGA and RGD-M13 phage/PLGA matrices. As shown in Figure 5, the proliferation of fibroblasts was increased in a time-dependent manner. However, fibroblasts cultured on the RGD-M13 phage/PLGA matrices showed the highest proliferation as compared with that on pure PLGA matrices and even the control. These results implied that the RGD peptides effectively promote not only initial attachment but also proliferation of fibroblasts to the matrices. In addition, proliferation of both cell lines and primary cells was increased. These results correspond well with previous studies. It has been reported that RGD-functionalized substrates are able to promote cell attachment [34, 35]. It is suggested that RGD peptides enhance cellular behaviors regardless of the cell types.Figure 4


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)

Cell attachment of L-929 cells, HDFs and HT-1080 cells on RGD-M13 phage/PLGA nanofiber matrices. Cell attachment was determined by a CCK-8 assay. Data are expressed as mean ± SD based on at least duplicate observations from three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: Cell attachment of L-929 cells, HDFs and HT-1080 cells on RGD-M13 phage/PLGA nanofiber matrices. Cell attachment was determined by a CCK-8 assay. Data are expressed as mean ± SD based on at least duplicate observations from three independent experiments.
Mentions: We examined cellular behaviors of fibroblasts on the RGD-M13 phage/PLGA nanofiber matrices. In this study, three types of fibroblast were used: murine fibroblast cell lines (L-929 cells), primary human dermal fibroblasts (HDFs), and human fibrosarcoma cell lines (HT-1080 cells). The cell viability was measured to assess a cell attachment when cells had been seed on the matrices for 6 hours. As shown in Figure 4, fibroblasts showed the highest adhesion where they were cultured on RGD-M13 phage/PLGA matrices regardless of cell types. As mentioned above, RGD peptides contained the matrices have contributed to the enhancement of cell adhesion. In contrast, cell adhesion on pure PLGA matrices was decreased because the PLGA matrices did not contain any materials which could improve cell adhesion. In addition, we evaluated the proliferation of fibroblasts on pure PLGA and RGD-M13 phage/PLGA matrices. As shown in Figure 5, the proliferation of fibroblasts was increased in a time-dependent manner. However, fibroblasts cultured on the RGD-M13 phage/PLGA matrices showed the highest proliferation as compared with that on pure PLGA matrices and even the control. These results implied that the RGD peptides effectively promote not only initial attachment but also proliferation of fibroblasts to the matrices. In addition, proliferation of both cell lines and primary cells was increased. These results correspond well with previous studies. It has been reported that RGD-functionalized substrates are able to promote cell attachment [34, 35]. It is suggested that RGD peptides enhance cellular behaviors regardless of the cell types.Figure 4

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