Experimental Study On Class â…¢ Furcation Defects Repaired With Autologous Periosteal Cells Transplantation In Beagle Dogs

PartⅠ:Effects of DMEM and PRMI1640 media on the biological behavior of periosteal cells from Beagle dogsObject: To investigate the effect of two media: DMEM and PRMI1640 on the behavior of periosteal cells from Beagle dogs, and provide the more ideal culture medium for periodontal tissue-engineering. Methods: Periosteum tissues derived from adult Beagle dogs were harevested, growing in DMEM and RPMI1640 mediums separately with modified tissue culture method. Then, the cell growth rate of periosteum tissues, proliferation speed, alkaline phosphate(ALP) and mineralized nodules of passaged cells cultured in vitro were observed. Finally, the effects of DMEM and RPMI 1640 culture medium on periosteal cells cultured in vitro were compared. Results: The cell growth rate of periosteum tissues, the proliferation speed, ALP and mineralized nodules in DMEM medium were better than in RPMI1640 medium. Conclusion: The culture medium DMEM may be more suitable for dog periosteal cells cultured in vitro.PartⅡ: Effect ofβ-tricalcium phosphate ceramic on the biological behavior of periosteal cells from Beagle dogsObject: to evaluate the effect ofβ-TCP on proliferation and differentiation of periosteal cells. Methods: periosteal cells isolated from Beagle dogs were cultured by tissue culture method in vitro. Cells were transferred in culture, and inoculated onto the surface ofβ-TCP. The proliferation and differentiation of periosteal cells were measured through MTT,flow cytometry (FCM) and alkaline phosphate (ALP) kit, and cellular morphology was examined by scanning electronic microscope. Results:periosteal cells could well adhered to and proliferated on the surface ofβ-TCP, and could have a higher alkine phosphatase activity.Conclusion:β-TCP has good biocompatibility and may be a acceptable scaffold for periodontal tissue engineering.PartⅢ: Periodontal regeneration following transplantation of periosteal cells into classⅢfurcation defects in Beagle dogsObject:to evaluate the healing of Beagle dogs classⅢfurcation defects following guided tissue regeneration and transplantation of periosteal cells. Methods:ClassⅢfurcation defects were created in 24 premolars of 4 adult Beagle dogs, gutta-percha was left in place for six weeks to maintain the depth of the defects and promote plaque accumulation. Each furcally involved premolar was then assigned to one of three treatments: Group A (e-PTFE), Group B(e-PTFE plusβ-TCP), Group C ( e- PTFE plusβ-TCP and periosteal cells). This resulted in eight total sites per treatment group. Twelve weeks after treatment and after no oral hygiene, tissue blocks of the mandible were taken for histometric analysis to access parameters of periodontal regeneration. Two teeth per treatment group were excluded from analysis because serious gingival recession or furcal perforation, and only six teeth per treatment were included in analysis. Results: Histometric analysis demonstrated, for group A, a surperior length of new periodontal ligament (3.71±1.82 mm verses 2.34±0.47 mm for group B, and verses 1.33±0.31 mm for group C), and a superior area of new bone (6.21±2.18 mm2 verses 3.71±2.00 mm2 for group B, and verses 2.35±1.26 mm2 for group C)(P<0.05). Conclusion: With the limit of the study, the transplantation of periosteal cells combined with GTR maybe a favarable way to regenerate periodontal defect.