Figure 8: Mkx-induced mesenchymal stem cells promote functional AF regeneration. | Nature Communications

Figure 8: Mkx-induced mesenchymal stem cells promote functional AF regeneration.

From: Mohawk promotes the maintenance and regeneration of the outer annulus fibrosus of intervertebral discs

Figure 8

(a) Strategy for assessing type I collagen synthesis of C3H10T1/2 cells induced by Venus (C3H10T1/2-V or Venus) and Venus-Mkx (C3H10T1/2-VM or Venus-Mkx) after outer AF (OAF) transplantation. (b) OAF before creating a transplantation site. Scale bar, 200 μm. (c) OAF after creating a transplantation site. Scale bar, 200 μm. (d) Cut surface of intervertebral disc (IVD) after creating a transplantation site. Yellow arrow, transplantation site. Scale bar, 200 μm. (e) Schema of the modified tail-looping model. Black arrow, transplanted site. (f,h) Haematoxylin and eosin staining of the transplantation sites of the OAF (f: C3H10T1/2-V; h: C3H10T1/2-VM). * The area in which the collagen gel was dissolved. ** Newly synthesized tissue. *** OAF. Scale bars, 50 μm. (g,i) IHC of these products (g: C3H10T1/2-V; i: C3H10T1/2-VM). Blue, Hoechst; green, anti-GFP. Scale bars, 50 μm. (j,k) Images of transmission electron microscopy (j: C3H10T1/2-V; k: C3H10T1/2-VM). Scale bars, 500 nm. (l) The calculated mean of the diameter of 100 collagen fibrils from C3H10T1/2-V and C3H10T1/2-VM cells. The mean diameter of 100 collagen fibrils was calculated: for C3H10T1/2-V cells, mean diameter=53.72 nm (s.d.=7.281 nm); for C3H10T1/2-VM cells, mean diameter=83.54 nm (s.d.=16.34 nm); for the intact OAF of C3H/HeSlc mice, mean diameter=77.74 nm (s.d.=24 nm). Error bars represent the s.d. **P<0.01. (m,n) Representative images of IVDs in each group. Safranin O fast green staining. (m) Whole images of each group. Transplantation sites of C3H10T1/2-V or C3H10T1/2-VM cells are on the left side in the images. Scale bars, 300 μm. (n) Images with a focus on the transplanted side of the AF. Scale bars, 100 μm. (o) Histological grading score of the AF using the Nishimura system. The mean average grade: control=2.4 (s.d.=0.5164); Venus=4 (s.d.=0.4082); Venus-Mkx=2.2 (s.d.=0.7528). **P<0.01. Statistical differences were assessed with Student’s t-test. GFP, green fluorescent protein.

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