The Experiment Research Of Repairing The Defect Of Cervical By Platelet-rich Plasma Gel Composite Materials

Objective: To investigate the capacity of repairing cervical bone defect of beagle dogs with platelet-rich plasma gel composite materials after being osteogenic induced , so as to provide experimental basis for establishing tissue-engineered bone and repairing cervical defects.Method: (1) Separation, purification, amplification of the beagle bone marrow mesenchymal stem cells, and preparation of platelet-rich plasma gel. (2) Choose the 3rd generation bone marrow mesenchymal stem cells respectively combine with platelet-rich plasma gel / beta-tricalcium phosphate composite scaffold material, simply beta-tricalcium phosphate composite scaffold and cultured with osteogenesis induced in vitro for 1 week. (3) Produced Beagle cervical bone defect model by anterior operative approach, and combined the prepared bone marrow mesenchymal stem cells with tricalcium phosphate alone enhanced stent (â… group), canine autologous bone (â…¡group), BMSCs compound strengthen the tricalcium phosphate / platelet-rich plasma gel scaffold (â…¢group) implanted cervical defect respectively. Then observe the tissue reaction around the implant, partial defect repair and bone formation and surface changes. (4) X-ray inspection was used at 4,8,12 week. Observe any changes in X-ray film. (5)Obtain the sample at 4,8,12 weeks and use HE staining and immunohistochemical to observe the osteocalcin protein expression. (6) RT-PCR was used to detect the expression of mRNA regarding OCN, ALP, CollA1 in the body by each groups at 4,8,12 week.Result: (1) There was no systemic and local inflammatory and toxicity response in each group of complex materials which implanted in beagle dogs with cervical bone defects,and the wound healing in each group was well. No significant inflammation and rejection occurred in implant sites. (2) The postoperative X-ray at 4,8,12 weeks displayed that groupâ…¡and groupâ…¢reached bone healing in 12 weeks, but the group I did not show complete absorption, bone healing, the materials - bone interface clear, material negative impact peripheral. (3) HE staining and osteocalcin protein immunohistochemical staining showed that endochondral bone emerged in each group, and the expression of enhanced tricalcium phosphate / autologous platelet-rich plasma gel composite material was stronger than the group of tricalcium phosphate group in 4,8,12 weeks. (4) The OCN, ALP and the mRNA expression of CollA1 emerging bands could be seen in 4,8,12 weeks. Compared the purpose band and the reference strip gray value of of each group, the expression of the groupâ…¡and groupâ…¢were stronger than group I in 8,12 weeks .there is no any significant difference between groupsâ…¡andâ…¢.Conclusion: Platelet-rich plasma gel / beta-tricalcium phosphate composite scaffold is an ideal material for bone tissue engineering scaffold material which has a satisfactory effect especially in histocompatibility, degradation rate and the capacity of osteogenics. Platelet-rich plasma gel / beta-tricalcium phosphate composite scaffold material can effectively promote osteoblastic effect of BMSCs after revulsant in vivo, in which osteoblastic phenotype are stably expressed . Platelet-rich plasma gel / beta-tricalcium phosphate composite scaffold material can effectively repair the bone defect of cervical in dogs which provide a experimental and theoretical basis for tissue engineering to repair bone defects in the cervical spine.