E<i>x-vivo</i> diaphragm muscle function at 31–33 days of age.

<p>2-Way ANOVA found no significant differences for peak twitch tension, time to peak twitch tension or time to ½ relaxation tension (p>0.05) between the four treatment groups. No effect of sex was observed for any of the above parameters (p>0.05). Values are means ± SEM; n = 5/group.</p

Postnatal growth for each treatment group from birth to 33±2 days.

<p>3-way repeated measures ANOVA showed a significant effect of age (p<0.05) in all treatment groups. No significant effects of birth or diet were found (p>0.05). Values are means ± SEM; n ≥6/group.</p

Mean cross-sectional area of the different fibre types in the spiny mouse diaphragm at 33 days of age for the four treatment groups.

<p>Values are means ± SEM; n = 5/group. * indicates significant difference to all other groups.</p

The number of dams pups, and the survival rates for each treatment group immediately after birth.

<p>The number of dams pups, and the survival rates for each treatment group immediately after birth.</p

Mean proportions of the different fibre types present in the spiny mouse diaphragm at 33 days of age for the four treatment groups.

<p>Values are means ± SEM; n = 5/group. * indicates significant difference to all other groups.</p

Microglial response to graphene-free or—inclusive PCL scaffold implantation at Week 1 and Week 3.

<p>(A) Image shows an overview of microglial infiltration into the outer shell of a gP6 scaffold at Week 3, and gP6 at high magnification at (B) Week 1 and (C) Week 3. (D) Graphene-free P6 scaffold at Week 3 (Green: Iba1⁺ microglia cells, blue: DAPI stained nucleus). (E) Microglial profile across the tissue/scaffold interfaces (*** p<0.001, n = 4). All brain tissue sections were collected on the transverse plane. Scale bar for (A) represents 100 μm; Scale bar for (B, C and D) represents 20 μm. Error bar in (E) shows standard error of the mean.</p

Microstructure of electrospun PCL microfibers and polyelectrolyte modified fibers with and without graphene.

<p>SEM images showing the microstructure of (A, B) PCL microfibers, (C) P6 and (D) gP6 modified microfibers. Partially aligned smooth microfiber morphology was revealed in all images at different magnifications.</p

Astrocyte morphology and infiltration at different time points following H6, P6, gP6, gH6 scaffolds implantation.

<p>Astrocyte/gP6 scaffolds interaction at Week (A) 1, (B) 3 and (C) 7; Astrocytes/P6 scaffolds interaction at Week (E) 3 and (F) 7; Astrocyte process infiltration into (H) gH6 at Week 3 and (I) H6 at Week 7. (D, G) detailed astrocyte morphology of the dash-box indicated area in (B, I) respectively. Green: GFAP positive astrocytes, blue: DAPI stained nucleus, red: surface functionalized scaffolds. * indicates astrocytes that bridge a gap between two scaffold layers in (D). All brain tissue sections were collected on the transverse plane. Scale bar for (D, G) represents 20 μm; for all other images represents 50 μm.</p

GFAP expression at different time points in the outer layer of scaffolds and in adjacent tissue.

<p>(150 μm from tissue/scaffold boundary) in terms of GFAP⁺ astrocytes occupied pixel percentage (pixel%). * (p<0.05, n = 4) indicate significant difference in GFAP expression between Week 3 and 7, within scaffold or tissue for gP6 implants. Error bar shows standard error of the mean.</p

Neuroblast migration/integration along/with gP6 implants at Week 3.

<p>Immunostaining images show DCX⁺ neuroblast (red) and nuclei (DAPI, blue) of tissue sections at the transverse plane. Schematic coronal brain section indicates the gP6 implantation track and locations of A-C, D-E tissue sections respectively. gP6 implant boundary is marked by dotted line. (A) gP6 implant in LV, bottom right part of the outermost scaffold layer is in direct contact with SVZ. Insert enlarges the box region showing neuroblast process across the entire scaffold layer. (B) Neuroblasts migrate along the scaffold surface/inter-layer gap and (C) processes infiltrating into the scaffold layers. Neuroblasts are identified in deeper sections of the gP6 implants either (D) within the scaffold or (E) close to implant in the LSV. LV: lateral ventricle, LSI: lateral septal nucleus, intermediate part, LSV: lateral septal nucleus, ventral part. Scale bars represent 20 μm for all images.</p