Regeneration Of Alveolar Bone With Tissue Engineered Bone: An Experimental Study Of Dogs

PurposeUsing the BMSCs-Bio-Oss tissue engineered bone which was constructed by BMSCs as seed cells and the Bio-Oss calf inogenic bone grains as scaffold materials to approach the feasibility of applying the tissue engineering combined with Bio-Gide collogen membrane to promote canine alveolar bone to regenerate.Methods1. Gybrid canine BMSCs were dissociated, cultivated, bone formation induced and amplificated, and then the third passage BMSCs were compounded with Bio-Oss calf inogenic bone grains to construct tissue engineered bone.2. BMSCs were observed by inverted phase contrast microscope during the course of being cultivated.3. The second bicuspid teeth were removed on preoperative three months, and three-wall bone defects were made artificially when operating. Then the twelve bone defect districts which were made artificially were divided into two groups. Experimentation groups were implanted tissue engineered bone which was constructed by BMSCs and Bio-Oss calf inogenic bone grains combined with Bio-Gide collogen membrane and then the gums had interrupted sutured; Control groups were merely implanted Bio-Oss grains combined with Bio-Gide collogen membrane and then the gums had interrupted sutured; Experimentation groups and control groups were both set periodontal packs after being sutured, and both had routine anti-inflammatory therapy at the 3^rd days after operation and the periodontal packs were demolished at the 14^th days after operation.4. Carrying out the imageology examination at the preoperative moment and at the second, forth, eighth week separately after operation and histology examination at the eighth week after operation to appraise the effect of canine alveolar bone regeneration by both HE and Masson dyeing.Results1. None of the experimental animals was dead and the drink and food of all experimental animals were normal. The cuts at the bone defect districts healed well and had no flare,infection and graft discarding.2. The tenth day of the BMSCs showed swirl shape to line up and spread out the whole culture flask ,which and fused to 70%～80% which were observed by inverted phase contrast microscope.3. The X-ray images of experimental groups and control groups at the preoperative moment showed that the tooth extraction wounds of both groups had completely bony healed and had the same bone density4. The X-ray images of experimental groups and control groups at the second week after the operation had no differences .At the fourth week after operation, experimental groups had new bone formation, the shade grew light at the bone defect districts. The normal bone trabeculae construction could be observed and the Bio-Oss grains were absorbed partly observed by X—ray. And the bone defect districts of control groups had little new bone formation and the edges were unsharped; at the eighth week after operation, the new bone mass of experimental groups increased obviously, and the Bio-Oss grains were absorbed completely. The bone defect districts were covered by normal bone trabeculae and their density were similar to that of surrounding bone. The bone defect districts were dense and the shade vanished. The shade of control groups grew lightl and normal bone trabeculae were observed.5. The histological section examination at the eighth week after operation showed that experimental groups constructed normal calcified bone construction and collogen membrane structure had vanished. The bone defect districts were filled with new alveolar bone, and bone trabeculae were small, multi-space and showed floccus shape. The calcification effect of new alveolar bone surpassed control groups obviously. The collogen were dyed red by Masson, and their calcifications were similar to normal alveolar bone. The bone defect districts were not full of new alveolar bone, and bone trabeculae were small, multi-space and in disorder at control groups. Athough it set up normal bone construction, its calcification degree was extreme low and was colllogen bone architecture basically with collogen dyed blue by Masson.ConclusionIt was feasible that to use the BMSCs-Bio-Oss tissue engineered bone which was constructed by BMSCs as seed cells and the Bio-Oss calf inogenic bone grains as scaffold materials to promote the generation of canine alveolar bone, and the growth velocity of the alveolar bone of experimental group surpass control group obviously.