Fabrication of an electrospun nanofibrous scaffold for use in the field

of tissue engineering

By: Shannon Daily & Tyler Crawford

Purpose To create a polycaprolactone mesh

which enables cell activity and seeks to eventually provide an application in the field of tissue engineering toward biomimetic skin graft.

Progress since last meeting Spun 8 more meshes

Visited Janelia Farm and used SEM

Details on cell procedure

Began cell work

Electrospun Meshes Mesh 5 & 6 Spun on 3/1 15 kV 15 cm from tip of pipette to collector

plate Pasteur pipette used Glass slide down middle

Electrospun Meshes

Mesh 7› 20 kV› 15 cm› No glass slide

Mesh 8› 20 kV› 20 cm› Glass slide

Mesh 7 & 8 Spun on 3/11 15 wt.% solution from 3/9

Electrospun Meshes

Mesh 9› 15 kV› 15 cm› Glass slide

Mesh 10› 20 kV› 15 cm› Glass slide

Mesh 9 & 10 Spun on 3/17 10 wt.% solution from 3/15

Electrospun Meshes

Mesh 11› 15 kV› 15 cm› Glass slide

Mesh 12› 20 kV› 15 cm› Glass slide

Mesh 11 & 12 Spun on 3/23 20 wt.% solution from 3/21

Chitosan Solution Created PCL/chitosan solution to begin

electrospinning 15 wt.% PCL, 1 wt.% chitosan

› 3 g PCL, .2 g chitosan, 16.8 g acetic acid

SEM ImagesMesh 4

SEM ImagesMesh 4

SEM ImagesMesh 6

SEM ImagesMesh 6

SEM ImagesMesh 6

Conclusion from SEM Beading is still a problem

› Sections of fibers and sections of beading Ideas to fix

› Increase the voltage› Decrease the concentration› Increase the distance

Comparison of fiber size› Mesh 1: 185 nm vs. Mesh 4&6: ~300 nm

Cell Work Meetings with Anu, Estelle, and Ms.

Curley› Determined basic procedure

Centrifuge thawed cells Add media to pellet of cells & resuspend Transfer to flask (in our case, tissue culture

plates) Allow cells to grow until about plate covered

about 80% Split to new containers

Cell Work Checked to see if mesh dissolves in

media› Placed small piece of mesh in 500 µL of

media› Let sit for 2 days- checking periodically› Mesh did not dissolve meaning cell growth

is possible

To be worked on: Use SEM on meshes created since last

time (7-12)› Different SEM?

Cell work› Begin with Mesh 4 (best fibers imaged)› Continue with other meshes

Begin spinning PCL/chitosan solutions

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