柚皮苷促进新型纳米陶瓷材料应用及诱导牙周再生机制的研究

The treatment of Periodontitis has been a difficult problem attracting much attention in the world. Periodontal tissue engineering is a research hotspot, which is still at the elementary stage yet. The following conclusions are arrived in the preliminary studies:(1) nano biphasic calcium phosphate (NBCP) was favorable for adsorption and accumulation of endogenous bone growth factors in circulation such as BMP4 and Cbfɑ1 after NBCP was implanted into the subcutis of beagle dogs;(2) the healing evidenced a new attachment apparatus after NBCP was implanted into periodontal alveolar bone defects; NBCP and MSCs implanted into periodontal alveolar bone defects promote periodontal regeneration;（3）drynaria extracts can promote proliferation and differentiation of marrow Mesenchymal Stem Cells（MSCs）in vitro. Based on the previous studies, this research , with the combination of Traditional Chinese Medicine’s theory of condition and growth of bone and periodontal tissue engineering ,is aimed to implant the coculture of MSCs and marerail in vitro in periodontal bone defects of beagle dogs following the injection of naringin to promote regeneration of alveolar bone. This paper uses various methods including micro-computed tomographic imaging, fluorescence microscope and in-situ hybridization ,real time PCR, with the combination of in-vivo and in-vitro experiment，to study the periodontal response and regeneration from the aspects of morphology and molecular biology, on the protein level and gene level. It explores the effects of the components from Traditional Chinese Medicine on optimization and bone-formation of domestic materials and develops a new point of conjunction for the treatment of periodontal diseases by Chinese Traditional Medicine combined with Western Medicine.

牙周病的治疗是全世界关注的难题，牙周组织工程技术是当前研究热点，但尚处于初步探索阶段。前期试验发现：①将一种新型纳米双相钙磷陶瓷埋入比格犬皮下组织，发现材料可以富集周围组织中Cbfa1和BMP4等关键骨生长因子。②将材料植入牙周缺损，显示有明显的牙周再生。材料与骨髓基质干细胞（MSCs）体外复合后植入牙周缺损，显示促进牙周再生。③骨碎补提取液可促进兔MSCs的增殖和成骨分化。 本课题在前期研究基础上进一步研究将材料与MSCs体外共培养后植入犬牙周缺损，局部应用柚皮苷以期促进牙周再生。本课题将采用显微CT、原位杂交等手段，从形态学、分子生物学、深入蛋白、基因水平观察牙周再生情况，探索柚皮苷能否促进MSCs的活性及生物学信号的增强，能否促进国产材料的优化和成骨、为中西结合治疗牙周病，开拓新结合点。

在正畸临床上，常见部分成人患者因牙周病、咬合创伤以及不当正畸力等导致牙周支持组织丧失、骨裂和骨穿孔等。对于伴发严重牙周组织丧失的错牙合畸形患者，促进缺失的牙周组织恢复或再生是保证正畸治疗顺利进行的前提。本课题在前期研究基础上，进一步将材料与MSCs体外共培养，探索不同浓度柚皮苷对人工骨的影响，浓度为2×10 -6 mol/L时细胞的活性最佳。共培养支架材料的电镜观察发现，细胞浓度为1×10 的7次方 个/mL时，与材料共培养6天时，细胞生长良好，全部覆盖材料表面。将细胞材料复合物植入犬牙周缺损，应用柚皮苷以期促进牙周再生，采用显微CT、原位杂交等手段，从形态学、分子生物学、深入蛋白、基因水平观察牙周再生情况，研究显示柚皮苷可促进MSCs的活性及生物学信号的增强，本实验为促进国产材料的优化和成骨、为中西结合治疗牙周病，开拓新结合点。

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