| Chronic Periodontitis (CP), a inflammatory disease which affect on tooth support tissue include gingiva, cementum, periodontal ligament and alveolar bone, is one of the two dominating oral diseases, CP is the main reason induce the tooth lost in adult. At present, the therapeutic efficacy of CP is just to control its symptom, to prevent its development, but hard to regenerate the lost tissue in periodontitis. Periodontal tissue regeneration provide a new way to regenerate the lost periodtontal tissue and seed cells, cell factor and scaffold are three elements in periodontal tissue regeneration. Zuk et al first found adipose derived stem cells at 2001, it was used in many kinds of tissue regeneration for it can be gain easily and can grow quickly. Adipose derived stem cells are not used widely in periodontal tissue regeneration; seed cells'differentiatio and transformation need suitable cell factor, platelet-rich plasma(PRP) which was concentration of platelet centrifuge from whole blood contain seven cell factors which were secrete by platelet, these cell factors relate to every wound healing, so PRP plays a very important role in seed cells'differentiatio and transformation. nano-HAp/Collagen(nHAC) is a new scaffold in periodontal tissue regeneration and it can promote its regeneration. This research investigate the beagle dog's adipose derived stem cells'isolation, culture, evaluation and its transformation to osteoblast, also we study the effect of platelet-rich plasma (PRP) on beagle dog's adipose derived stem cells when it differentiate into osteoblast, and the biocompatibility of beagle dog's adipose derived stem cells and nano-HAp/Collagen, to provide the experimental basis for suitable seed cells, cell factor and scaffold in periodontal tissue regeneration. At the same time, transplant the compound of adipose derived stem cells-PRP- nano HAp/Collagen to the experimental periodontist defect, so as to explore the effect of this kind of compound on periodontal tissue regeneration.Object: The aim of this study was to investigate the approach of beagle dog's adipose derived stem cells'culture, evaluation and its competence of differentiation into mature cells, to survey the effect of platelet-rich plasma (PRP) on beagle dog's adipose derived stem cells when it differentiate into osteoblast, and the biocompatibility of beagle dog's adipose derived stem cells with nano-HAp/Collagen, to explore the effect of the compound of adipose derived stem cells-PRP- nano HAp/Collagen in periodontal tissue regeneration. To investigate the feasibility of adipose derived stem cells as periodontal tissue regeneration'seed cell, to provide the experimental basis for suitable seed cells and scaffold in periodontal tissue regeneration.Method: Adipose derived stem cells were obtained by enzyme digestion, and immunohistochemistry S-P method was uesd to detect the CD44 and CD29 expression, to assess its stem cell characteristics. Then cultured the cells in DMEM with 10 mL/L FBS, Osteoblasts induced medium and Osteoblasts induced medium with PRP respectively. The result was observed with von kossa staining after 21 days, to observe the beagle dog's adipose derived stem cells'competence of differentiation into osteoblasts and the effect of platelet-rich plasma (PRP) on its differentiate into osteoblast. Adipose derived stem cells were cocultured with nano-HAp/Collagen, cell attachment was observed by scanning electron microscope after 7 days. Transplant the compound of adipose derived stem cells-PRP- nano HAp/Collagen to the experimental periodontist defect, Animal were killed at 4 weeks, 8 weeks and 12 weeks after surgery, to explore the effect of this kind of compound on periodontal tissue regeneration.Result: 1 Enzyme digestion can obtain adipose derived stem cells effectively, the source of mesenchyme stem cell can evaluate by CD44,CD29 staining;2 PRP has the best effect on the proliferation of adipose derived stem cells when the cell culture medium contains 1.5 times of whole blood concentration of platelet concentration3 Induced by osteoblasts induced medium, primary cultured stem cells can differentiated into osteoblasts, form calcified nodules, PRP can promote adipose derived stem cells to form more calcified nodules;4 Adipose derived stem cells can compound grow in the scaffold, adhere tightly and fully extend on nano-HAp/Collagen after 7 days cocultureConclusion: 1 Adipose derived stem cells can obtain convenient,have high survival rate and strong reproductive activity, can differentiated into osteoblasts with the contribution of osteoblasts induced medium, it can be used as a satisfactory seed cell in periodontal tissue regeneration;2 PRP can improve adipose derived stem cells'multiplication, enhance its differentiation to osteoblasts. PRP can be used as suitable cell factor3 nano-HAp/Collagen has favorable biocompatibility, can be used as scaffold in periodontal tissue regeneration4 Adipose derived stem cells-PRP- nano-HAp/Collagen is a excellent combination to promote periodontal regeneration.
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