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Electrospun silk fibroin fiber diameter influences in vitro dermal fibroblast behavior and promotes healing of ex vivo wound models.

Hodgkinson T, Yuan XF, Bayat A - J Tissue Eng (2014)

Bottom Line: Cell morphology was observed to depart from a stellate morphology with numerous cell -fiber interactions to an elongated, fiber-aligned morphology with interaction predominately with single fibers.The re-epithelialization of ex vivo wound models was significantly improved with the addition of BMSF electrospun scaffolds, with migratory keratinocytes incorporated into scaffolds.The results may aid in the development of effective BMSF electrospun scaffolds for wound healing applications.

View Article: PubMed Central - PubMed

Affiliation: Plastic & Reconstructive Surgery Research, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK ; School of Chemical Engineering and Analytical Science, University of Manchester, Manchester, UK.

ABSTRACT
Replicating the nanostructured components of extracellular matrix is a target for dermal tissue engineering and regenerative medicine. Electrospinning Bombyx mori silk fibroin (BMSF) allows the production of nano- to microscale fibrous scaffolds. For BMSF electrospun scaffolds to be successful, understanding and optimizing the cellular response to material morphology is essential. Primary human dermal fibroblast response to nine variants of BMSF scaffolds composed of nano- to microscale fibers ranging from ~250 to ~1200 nm was assessed in vitro with regard to cell proliferation, viability, cellular morphology, and gene expression. BMSF support of epithelial migration was then assessed through utilization of a novel ex vivo human skin wound healing model. Scaffolds composed of the smallest diameter fibers, ~250 -300 nm, supported cell proliferation significantly more than fibers with diameters approximately 1 μm (p < 0.001). Cell morphology was observed to depart from a stellate morphology with numerous cell -fiber interactions to an elongated, fiber-aligned morphology with interaction predominately with single fibers. The expressions of extracellular matrix genes, collagen types I and III (p < 0.001), and proliferation markers, proliferating cell nuclear antigen (p < 0.001), increased with decreasing fiber diameter. The re-epithelialization of ex vivo wound models was significantly improved with the addition of BMSF electrospun scaffolds, with migratory keratinocytes incorporated into scaffolds. BMSF scaffolds with nanofibrous architectures enhanced proliferation in comparison to microfibrous scaffolds and provided an effective template for migratory keratinocytes during re-epithelialization. The results may aid in the development of effective BMSF electrospun scaffolds for wound healing applications.

No MeSH data available.


Related in: MedlinePlus

Representative scanning electron micrographs of BMSF scaffolds and fiber diameter distribution histograms. Red curves indicate normal distributions calculated using the means and standard deviations of each fiber set.BMSF: Bombyx mori silk fibroin.
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fig1-2041731414551661: Representative scanning electron micrographs of BMSF scaffolds and fiber diameter distribution histograms. Red curves indicate normal distributions calculated using the means and standard deviations of each fiber set.BMSF: Bombyx mori silk fibroin.

Mentions: To produce fibers with varying diameters, the BMSF-PEO solutions were spun under various electrospinning process parameter conditions. The resultant fibers were analyzed, after insolubilization treatment by submersion in 100% (v/v) methanol for 10 min at room temperature, through scanning electron microscopy (SEM) of gold sputter-coated samples (FEI Quanta 200 (E)SEM + energy-dispersive X-ray (EDX); FEI, USA). Fibers to be used in this study were selected on the basis of their fiber diameter and standard deviations. Fiber measurements were conducted using ImageJ Software v1.45 (NIH, USA). To calculate diameter, 50 fiber diameters were measured at random to obtain mean diameter and standard deviation. For histograms of fiber diameter distribution, numbers of fiber diameters were converted into percentage total values and plotted against grouped fiber diameter. The mean fiber diameters are shown, along with standard deviations (Figure 1; Table 1).


Electrospun silk fibroin fiber diameter influences in vitro dermal fibroblast behavior and promotes healing of ex vivo wound models.

Hodgkinson T, Yuan XF, Bayat A - J Tissue Eng (2014)

Representative scanning electron micrographs of BMSF scaffolds and fiber diameter distribution histograms. Red curves indicate normal distributions calculated using the means and standard deviations of each fiber set.BMSF: Bombyx mori silk fibroin.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4221927&req=5

fig1-2041731414551661: Representative scanning electron micrographs of BMSF scaffolds and fiber diameter distribution histograms. Red curves indicate normal distributions calculated using the means and standard deviations of each fiber set.BMSF: Bombyx mori silk fibroin.
Mentions: To produce fibers with varying diameters, the BMSF-PEO solutions were spun under various electrospinning process parameter conditions. The resultant fibers were analyzed, after insolubilization treatment by submersion in 100% (v/v) methanol for 10 min at room temperature, through scanning electron microscopy (SEM) of gold sputter-coated samples (FEI Quanta 200 (E)SEM + energy-dispersive X-ray (EDX); FEI, USA). Fibers to be used in this study were selected on the basis of their fiber diameter and standard deviations. Fiber measurements were conducted using ImageJ Software v1.45 (NIH, USA). To calculate diameter, 50 fiber diameters were measured at random to obtain mean diameter and standard deviation. For histograms of fiber diameter distribution, numbers of fiber diameters were converted into percentage total values and plotted against grouped fiber diameter. The mean fiber diameters are shown, along with standard deviations (Figure 1; Table 1).

Bottom Line: Cell morphology was observed to depart from a stellate morphology with numerous cell -fiber interactions to an elongated, fiber-aligned morphology with interaction predominately with single fibers.The re-epithelialization of ex vivo wound models was significantly improved with the addition of BMSF electrospun scaffolds, with migratory keratinocytes incorporated into scaffolds.The results may aid in the development of effective BMSF electrospun scaffolds for wound healing applications.

View Article: PubMed Central - PubMed

Affiliation: Plastic & Reconstructive Surgery Research, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK ; School of Chemical Engineering and Analytical Science, University of Manchester, Manchester, UK.

ABSTRACT
Replicating the nanostructured components of extracellular matrix is a target for dermal tissue engineering and regenerative medicine. Electrospinning Bombyx mori silk fibroin (BMSF) allows the production of nano- to microscale fibrous scaffolds. For BMSF electrospun scaffolds to be successful, understanding and optimizing the cellular response to material morphology is essential. Primary human dermal fibroblast response to nine variants of BMSF scaffolds composed of nano- to microscale fibers ranging from ~250 to ~1200 nm was assessed in vitro with regard to cell proliferation, viability, cellular morphology, and gene expression. BMSF support of epithelial migration was then assessed through utilization of a novel ex vivo human skin wound healing model. Scaffolds composed of the smallest diameter fibers, ~250 -300 nm, supported cell proliferation significantly more than fibers with diameters approximately 1 μm (p < 0.001). Cell morphology was observed to depart from a stellate morphology with numerous cell -fiber interactions to an elongated, fiber-aligned morphology with interaction predominately with single fibers. The expressions of extracellular matrix genes, collagen types I and III (p < 0.001), and proliferation markers, proliferating cell nuclear antigen (p < 0.001), increased with decreasing fiber diameter. The re-epithelialization of ex vivo wound models was significantly improved with the addition of BMSF electrospun scaffolds, with migratory keratinocytes incorporated into scaffolds. BMSF scaffolds with nanofibrous architectures enhanced proliferation in comparison to microfibrous scaffolds and provided an effective template for migratory keratinocytes during re-epithelialization. The results may aid in the development of effective BMSF electrospun scaffolds for wound healing applications.

No MeSH data available.


Related in: MedlinePlus