The Study On Effect Of Tissue Engineering Materials KLD-12 Polypeptide /TGF-β1 Nanofiber Gel Composite BMSCs To Treatment Of Disc Degeneration

Objective: To discuss the effect of tissue engineering materials KLD- 12 polypeptide /TGF- beta 1 composite with BMSCs to treat intervertebral disc degeneration.Methods: To choose 40 healthy New Zealand white rabbit, randomly divided into A, B, C, D, E groups, including A, B, C, D in model groups(L3-4、 L4-5、L5-6). After two weeks, we will implant 40 ul something into degeneration of intervertebral disc in situ gel, including KLD- 12 ploypeptide/TGF- beta/BMSCs gel, physiological saline, KLD- 12 ploypeptide /BMSCs gel, KLD- 12 ploypeptide / TGF-β1.The group E do not do anything. After transplantation 4 weeks, 8 weeks, 12 weeks and 24 weeks, we randomly choose 2 rabbit to inspect X-ray and MRI imaging in each group. Then observation intervertebral disc changes between groups by organizations observation, HE staining, Toluidine blue staining, semi-quantitative RT-PCR detection of intervertebral disccollagen typeⅡ and Aggrecan m RNA expression in the rabbit intervertebral disc specimens.Results:The X-ray examination showed, % DHI compared with the A group at each time point, B, C, D group of 4 weeks,8 weeks, 12 weeks, 24 weeks were decreased, the difference was statistically significant(P < 0.05). Compared with the A group, the T2 weighted signal intensity of MRI in C and D were lower, B was lowest, E was higher. Accordin, we compared it with the A: there were statistically significant(P < 0.05). Gross observation, group A can be seen newborn nucleus pulposus tissue; group B central nucleus pulposus tissue loss, showing a pale yellow fibrous tissue hyperplasia; group C intervertebral slightly narrowed, the central nucleus area visible white fibrous tissue hyperplasia, texture tough, peripheral annulus structure slightly disordered; D group showed a small amount of newborn nucleus pulposus tissue.H&E staining,ccording to the histological grade of disc degeneration: group A 1~2 grade, group B 4~5 grade, group C 3~4 grade, group D,2~3 grade, group E,0grade. Toluidine blue,group A of nucleus pulposus depth toluidine blue staining, Group B nucleus pulposus toluidine blue stained portion of the nucleus pulposus tissue loss, not colored, local lightly stained; group C and D in the nucleus pulposus toluidine blue moderate of staining; Group E pulposus depth toluidine blue staining. Aggrecan m RNA, group A ~ group E were expressed, group A compared with group B, C, D a significant increase in the expression of various points of time, the difference was statistically significant(P <0.05); group A compared with group E, expression was significantly lower, the difference was statistically significant(P <0.05). CollagenⅡm RNA, group A compared with group B, C, D a significant increase in the expression of various points of time, the difference was statistically significant(P <0.05); group A compared with group E, expression was significantly lower, the difference was statistically significant(P <0.05).Conclusion: KLD-12 peptide / TGF-β1 complex BMSCs transplantation has some repair and regeneration effect to degenerative disc.