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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: 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.

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

MTT analysis of cell proliferation on BMSF fibrous scaffolds. Cell proliferation in 256 and 304 nm diameter fibrous scaffolds is significantly enhanced during the culture period (*p < 0.05; **p < 0.001). Values are means of five independent experiments performed in triplicate with five different PHDF populations.BMSF: Bombyx mori silk fibroin; PHDF: primary human dermal fibroblast; OD: optical density.
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fig3-2041731414551661: MTT analysis of cell proliferation on BMSF fibrous scaffolds. Cell proliferation in 256 and 304 nm diameter fibrous scaffolds is significantly enhanced during the culture period (*p < 0.05; **p < 0.001). Values are means of five independent experiments performed in triplicate with five different PHDF populations.BMSF: Bombyx mori silk fibroin; PHDF: primary human dermal fibroblast; OD: optical density.

Mentions: Through control of electrospinning solution properties and process parameters,13 scaffolds of uniform BMSF fibers were generated and scaffolds of increasing fiber diameter were chosen for analysis from 256 ± 30 to 1214 ± 321 nm (Figure 1). To assess the effects of electrospun BMSF fiber diameter on cell proliferation in culture, MTT assays were performed to quantify viable cell number up to 14 days (Figure 3). Initial proliferation in the first 48 h of culture was comparable between scaffolds. After 72 h, PHDF proliferation on BMSF scaffolds with fibers ~250–300 nm in diameter was increased in comparison with larger diameter fibers. After 1 week, there is a highly significant difference in cell proliferation between fibers below 304 nm in diameter and those 400 nm and above, with a general trend of decreasing proliferation for increasing fiber diameter. Interestingly, the largest increase in cell number is observed between cells cultured on scaffolds ~300 and ~400 nm. The proliferation in scaffolds between ~850 and ~1200 nm was comparable during culture. Optimal cell proliferation was seen in scaffolds with the smallest fiber diameters, approximately 250–300 nm (p < 0.001) (Figure 3).


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)

MTT analysis of cell proliferation on BMSF fibrous scaffolds. Cell proliferation in 256 and 304 nm diameter fibrous scaffolds is significantly enhanced during the culture period (*p < 0.05; **p < 0.001). Values are means of five independent experiments performed in triplicate with five different PHDF populations.BMSF: Bombyx mori silk fibroin; PHDF: primary human dermal fibroblast; OD: optical density.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

fig3-2041731414551661: MTT analysis of cell proliferation on BMSF fibrous scaffolds. Cell proliferation in 256 and 304 nm diameter fibrous scaffolds is significantly enhanced during the culture period (*p < 0.05; **p < 0.001). Values are means of five independent experiments performed in triplicate with five different PHDF populations.BMSF: Bombyx mori silk fibroin; PHDF: primary human dermal fibroblast; OD: optical density.
Mentions: Through control of electrospinning solution properties and process parameters,13 scaffolds of uniform BMSF fibers were generated and scaffolds of increasing fiber diameter were chosen for analysis from 256 ± 30 to 1214 ± 321 nm (Figure 1). To assess the effects of electrospun BMSF fiber diameter on cell proliferation in culture, MTT assays were performed to quantify viable cell number up to 14 days (Figure 3). Initial proliferation in the first 48 h of culture was comparable between scaffolds. After 72 h, PHDF proliferation on BMSF scaffolds with fibers ~250–300 nm in diameter was increased in comparison with larger diameter fibers. After 1 week, there is a highly significant difference in cell proliferation between fibers below 304 nm in diameter and those 400 nm and above, with a general trend of decreasing proliferation for increasing fiber diameter. Interestingly, the largest increase in cell number is observed between cells cultured on scaffolds ~300 and ~400 nm. The proliferation in scaffolds between ~850 and ~1200 nm was comparable during culture. Optimal cell proliferation was seen in scaffolds with the smallest fiber diameters, approximately 250–300 nm (p < 0.001) (Figure 3).

Bottom Line: 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.

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