1. Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized commercial reproduction of this slide is prohibited Supplemental PowerPoint Slides Effects of transplantation of hTIMP1-expressing bone marrow mesenchymal stem cells on the extracellular matrix of degenerative intervertebral discs in an in vivo rabbit model Yi Zhou, MDa,b, Guanjun Tu, MD, PhDa,*, Lin Cong, MDa, Zifeng Pei, MDb aDepartment of Orthopedics, First Hospital Affiliated with China Medical University, Shenyang 110001, China bDepartment of Orthopedics, Second Hospital Affiliated with Shenyang Medical College, Shenyang 110002, China

2. Figure 2. Comparison of tissues across groups. A. Example discs obtained from rabbits in each group showing anatomical and morphological differences between the groups. B. Hematoxylin and eosin stained sections (400×) demonstrating fewer and more irregularly shaped NP cells in animals from the PCon group relative to the h-TIMP-1 TgBT group.

3. Figure 3. Quantification of PG and collagen type II content. A. Spectrographic analysis revealed that PG content was significantly lower in the PCon group and BT group than in the h-TIMP-1 TgBT group (*P <.05). B. Likewise, spectrographic analysis revealed that collagen type II content was significantly lower in the PCon group and BT group than in the h-TIMP-1 TgBT group (*P <.05).

4. Figure 4. Protein expression in NP tissue specimens from each group after 12 weeks, with buffy color indicating positive staining. A. Type II collagen labeled sections. (400×) B. hTIMP-1 labeled sections. (400×)

5. Figure 5. Summary of data demonstrating successful elevation of hTIMP-1 expression in h-TIMP-1 TgBT group. A. hTIMP-1 solubility curve obtained after fluorescent Q- PCR. Note the unimodal curve, indicating that one PCR product was obtained. B. Comparison of hTIMP-1 mRNA expression between groups. There was elevated expression of hTIMP-1 mRNA in the h-TIMP-1 TgBT group (*P <.05 vs. BT group). C. Western blot for hTIMP-1 protein expression. We observed higher expression of hTIMP-1 protein in the h-TIMP-1 TgBT group. D. Quantitative analysis of hTIMP-1 protein expression. There was elevated expression of hTIMP-1 protein in the h-TIMP- 1 TgBT group (*P <.05 vs. BT group).

6. Figure 1. Digital radiographic analysis of the spinal columns revealed more severe sclerosis of the endplate of the intervertebral disc and shorter relative intervertebral space height (RISH) in the PCon group and BT group compared to the h-TIMP-1 TgBT group (*P <.05).