Angiogenesis after injection of VEGF-loaded PLGA/DC-Chol nanospheres.

<p>(A) Immunofluorescent staining for SM-α actin in injured spinal cords 4 weeks after injection. Scale bars indicate 100 μm at 100× and 20 μm at 400× (B) Quantification of arteriole density in injured spinal cords. *<i>p</i> < 0.05 vs. PBS, naked VEGF, or PEI/VEGF. #<i>p</i> < 0.05 vs. PBS.</p

(A) Cytotoxicity of pDNA-loaded nanospheres and PEI/pDNA complexes. mNSCs were cultured for 4, 24, or 48 h with the indicated polymer concentrations, and mitochondrial metabolic activity in mNSCs was measured using the MTT assay. <i>p < 0</i>.<i>05</i> for nanospheres (PLGA/DC-Chol or PLGA) vs. PEI at all concentrations and time points (n = 5) except for 10 and 20 μg at 4 h.

<p>Transfection efficiency in cultured mNSCs (B, C). (B) Luciferase gene expression in mNSCs transfected with pSV-Luc-loaded PLGA nanospheres, PLGA/DC-Chol nanospheres, or PEI /pSV-Luc complex. *<i>p</i> < 0.05.PEI/pLuci compare with naked, PLGA and PLGA/DC-Chol at 7 days after transfection. <i># p</i> < 0.05 PLGA/DC-Chol nanospheres compared with PLGA, PEI and naked at 14 days after transfection. **<i>p</i> < 0.05 PLGA nanospheres compared with PEI and naked at 14 days after transfection. (C) Luciferase mRNA expression 7 and 14 days after gene transfection.</p

Gene expression in the rat spinal cord.

<p>(A, B) Luciferase activity in the rat spinal cord 14 days after injection. *<i>p</i> < 0.05 compared with naked pLuci and PLGA/pLuci. # <i>p</i> < 0.05 compared with naked and PEI/pDNA, + <i>p</i> < 0.05 compared with PEI/pDNA. Duble immunofluorescent staining for (C) beta III tublin (red), luciferease (green) and betaIII tublin/luciferase merged cells (yellow) (D) GFAP (green), luciferase (red) and GFAP/luciferase merged cells (yellow), (E) Smooth muscle α-actin (red), luciferase (green) and SM α-actin/luciferase merged cells(yellow) in spinal cord. The arrows indicate cells of luciferase expression and beta III tublin positive cells, GFAP positive cells or sm-α actin positive cells. Scale bars indicate 50 μm.</p

Characterization of nanospheres.

<p>SEM images of (A) PLGA and (B) PLGA/DC-Chol nanospheres. (C) Size distribution of nanospheres. Zeta potential (mV) of (D) PLGA and (E) PLGA/DC-Chol nanospheres. (F) Agarose gel retardation analysis. Agarose gel electrophoresis of pDNA released from PLGA and PLGA/DC-Chol nanospheres. (G) Cumulative release of pDNA from PLGA and PLGA/DC-Chol nanospheres.</p

Axonal growth at lesion site and functional recovery 4 weeks after injury and injection.

<p>(A) Double immunostaining with GFAP (red) and NF (green). Scare bar indicates 100 μm. (B) Quantification of NF optical density of regenerated area. *p < 0.05 vs. PBS, naked VEGF, and PEI/VEGF. #p < 0.05 vs. PBS. (C) Effect of VEGF-loaded PLGA/DC-Chol nanospheres on recovery of locomotor function. BBB score was significantly greater in the PLGA/DC-Chol nanosphere group (n = 5) compared to PLGA/VEGF (n = 5), PEI/VEGF (n = 5), naked VEGF (n = 5), and PBS (n = 5) 3 and 4 weeks after injury. *p < 0.05 for PLGA/DC-Chol <i>vs</i>. PBS, naked VEGF, and PEI/VEGF groups.</p

Physiochemical characterization of PLGA/DC-Chol nanospheres.

<p>Physiochemical characterization of PLGA/DC-Chol nanospheres.</p

Additional file 5: Table S3. of Human-induced pluripotent stem cells generated from intervertebral disc cells improve neurologic functions in spinal cord injury

Lists of human embryonic stem cell (hESC)-enriched genes and chondrogenic genes shown in Fig. 2j (right)

Additional file 1: Table S5. of Human-induced pluripotent stem cells generated from intervertebral disc cells improve neurologic functions in spinal cord injury

List of the antibodies used in this study

Additional file 3: Table S1. of Human-induced pluripotent stem cells generated from intervertebral disc cells improve neurologic functions in spinal cord injury

DNA fingerprint analysis

The flexible neuronal conversion of malignant C6 glioma through the action of small molecules.

<p>(A and B) Representative images of C6 glioma before neural induction. (C) Representative fluorescence image of Tuj1-positive cells 7 days after neural induction of glioma with CHIR99021 (2 μM) and forskolin (10 μM). (D) Representative image of Tuj1/DAPI staining at 7 days after treatment of mouse embryonic fibroblasts with a combination of CHIR99021 (2 μM) and forskolin (10 μM). (E) Representative image of Tuj1-positive cells 7 days after treatment of glioma with a variety of concentrations of CHIR99021 and forskolin. Scale bars represent 500 μm (A), 50 μm (B), 100 μm (C), 100 μm (D), and 100 μm (E).</p