牙周组织工程中壳聚糖—丝素蛋白—磷酸三钙复合物的应用研究

发布时间：2019-03-26 12:36

【摘要】：牙周疾病是引起牙周组织缺损最终导致牙齿缺失的常见病因，如何有效防治牙周病所引起的组织缺损及功能障碍，成为口腔科医生亟待解决的问题。组织工程学理论和技术的飞速发展，为牙周组织再生研究提供了新的思路[1]。用组织工程学方法促进牙周再生已成为这一领域的研究热点之一。如何选择具有增殖分化潜能的理想种子细胞及符合组织工程要求的支架材料是组织工程研究的重要内容。目的：以犬牙周膜细胞（PDLCS）为种子细胞，壳聚糖-丝素蛋白-磷酸三钙复合物作为支架材料，，应用于牙周组织工程，探讨壳聚糖-丝素蛋白-磷酸三钙复合物的优越性。方法：选择1-2岁的健康雄性杂种犬6只，以每只犬下颌双侧第2、3、4前磨牙为实验牙，建立雄性杂种犬的牙周缺损模型；模型建立后，应用随机化法将其分为空白对照组、壳聚糖组、壳聚糖-丝素蛋白-磷酸三钙复合物组，每组2只犬。体外培养犬牙周膜细胞，取传至第4代细胞接种到支架材料上，并于2 h内分别植入犬下颌双侧第2、3、4前磨牙的牙周缺损处；术后8周处死动物，取实验处牙槽骨；经常规固定、脱钙、包埋后，HE染色，光镜下观察牙周组织再生情况。MTT法检测支架材料对狗PDLCs增殖的影响。所得各组数据使用SPSS17.0软件进行统计分析。结果：HE染色结果显示，壳聚糖-丝素蛋白-磷酸三钙复合支架材料用于牙周组织工程，其牙周间隙减小、成骨细胞数量增多较壳聚糖组和空白对照组更为明显。MTT法显示，壳聚糖/丝素蛋白/磷酸三钙复合支架对狗PDLCs的生长、增殖与单纯壳聚糖作为支架材料对照相比，差异无统计学意义（P 0.05）结论：以犬牙周膜细胞（PDLCs）为种子细胞，壳聚糖-丝素蛋白-磷酸三钙复合支架材料较单纯壳聚糖支架材料具有更强的组织相容性和再生能力，具有可行性。
[Abstract]:Periodontal disease is the common cause of periodontal tissue defect and tooth loss. How to effectively prevent and cure the tissue defect and dysfunction caused by periodontal disease has become an urgent problem for stomatologists to solve. The rapid development of tissue engineering theory and technology provides a new idea for the study of periodontal tissue regeneration. Tissue engineering to promote periodontal regeneration has become one of the research hotspots in this field. How to select ideal seed cells with proliferation and differentiation potential and scaffold materials which meet the requirements of tissue engineering is an important part of tissue engineering research. Aim: to investigate the advantages of chitosan-silk fibroin-tricalcium phosphate complex in periodontal tissue engineering by using canine periodontal ligament cell (PDLCS) as seed cell and chitosan-fibroin-tricalcium phosphate complex as scaffold material. Methods: six healthy male hybrid dogs aged 1-2 years were used as experimental teeth to establish periodontal defect model of male hybrid dogs with the 2nd, 3rd and 4th premolars of each dog's mandible as experimental teeth. After the model was established, the dogs were randomly divided into control group, chitosan-silk fibroin-tricalcium phosphate complex group (2 dogs in each group). Canine periodontal ligament cells were cultured in vitro and seeded into the scaffolds in passage 4, then implanted into the periodontal defects of canine mandibular premolars at the 2nd, 3rd and 4th premolars within 2 hours, and the animals were killed 8 weeks after operation and the alveolar bones were taken from the experimental sites. After routine fixation, decalcification, embedding and HE staining, the regeneration of periodontal tissue was observed under light microscope. The effect of scaffolds on the proliferation of dog PDLCs was detected by MTT method. The data were analyzed by SPSS17.0 software. Results: the results of HE staining showed that when chitosan-silk fibroin-tricalcium phosphate composite scaffold was used for periodontal tissue engineering, the periodontal space decreased and the number of osteoblasts increased more obviously than that of chitosan group and blank control group. The growth and proliferation of chitosan / silk fibroin / tricalcium phosphate composite scaffold on dog PDLCs were compared with that of chitosan as scaffold material. There was no significant difference (P 0.05). Conclusion: (PDLCs) is the seed cell of canine periodontal ligament cells. Chitosan-silk fibroin-tricalcium phosphate composite scaffolds have stronger histocompatibility and regeneration ability than pure chitosan scaffolds, so it is feasible to use chitosan-silk protein-tricalcium phosphate composite scaffolds.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08

【参考文献】