载苯妥英钠、硝苯地平、环孢素的PLGA缓释微球对牙周组织再生的作用

发布时间：2018-06-21 20:43

本文选题：SD大鼠 + 苯妥英钠　；参考：《武汉大学》2015年博士论文

【摘要】：第一部分大鼠药物性牙龈增生模型的建立实验目的：以SD雄性大鼠为实验对象,建立药物性牙龈增生的动物模型。实验方法：选择两种致药物性牙龈增生的代表药物,分别为抗癫痫药苯妥英钠和钙通道阻滞剂硝苯地平,给药方式为全身给药(口服),给药时间设定为25天、50天、75天,按不同给药时间对大鼠进行分组,对大鼠进行给药实验,每日给药一次,给药期结束后,处死实验鼠,收集上下颌骨及牙周组织,在体视显微镜下观察并测量牙龈厚度的变化,然后石蜡包埋切片,HE染色及免疫组化,并分析实验结果。实验结果：在苯妥英钠或硝苯地平长期口服给药下,SD雄性大鼠的牙龈表现出较为明显的增生(p0.01或p0.05),与对照组比较发现,实验组牙龈厚度增厚,牙龈上皮钉突伸长。结论：苯妥英钠和硝苯地平长期全身用药,可以诱导大鼠牙龈组织的药物性增生。第二部分苯妥英钠局部注射给药对大鼠牙龈的作用实验目的：在SD雄性大鼠上颌磨牙的牙龈局部注射致牙龈增生的药物苯妥英钠溶液,观察苯妥英钠对SD大鼠牙龈的作用情况。实验方法：SD雄性大鼠行进行全身麻醉,采用自身对照的方法,在大鼠上颌一侧的第一磨牙颊侧龈颊沟黏膜下注射苯妥英钠溶液,每日给药一次,给药剂量为20ul,间隔24小时,40天后处死实验鼠,收集大鼠上颌骨及牙周组织,观察并测量大鼠牙龈厚度的变化。实验结果：给药结束后收集分析数据结果显示,局部注射苯妥英钠溶液后,与对照组相比,大鼠实验组的牙龈并未出现增生(p0.05)。结论：以苯妥英钠溶液的形式进行局部注射给药不能诱导大鼠牙龈增生,有必要改进给药方法和剂型。第三部分载苯妥英钠、硝苯地平、环孢素的聚乳酸羟基乙酸(PLGA)缓释微球对牙周组织再生的作用实验目的：苯妥英钠、硝苯地平、环孢素是三大类致药物性牙龈增生药(抗癫痫药,钙通道拮抗剂,免疫抑制剂)的代表药,通过一种生物相容性材料PLGA作为载体,将这三种药物分别导入大鼠牙龈退缩模型中,以缓释给药的方式,检验三种药物,对牙龈退缩的治疗效果。实验方法：1.制备苯妥英钠、硝苯地平、环孢素三种药物的PLGA缓释微球；2.通过高效液相色谱法检验三种PLGA缓释微球在体外释放的药物成分、释放浓度及释放时间；3.在SD雄性大鼠上颌磨牙区制备牙龈退缩的动物模型；4.将三种载药微球导入制备好的牙龈退缩大鼠牙槽骨缺损处,2个月后,处死取材,在体视显微镜下观察牙龈增生情况,显微CT观察牙槽骨再生,大鼠上颌颌骨及牙周组织石蜡包埋切片,检测胶原纤维相关蛋白,成骨细胞、破骨细胞相关蛋白的表达。实验结果：1.三种药物的PLGA缓释微球制备成功,宏观形态及扫描电镜下形态良好,符合文献描述；2.高效液相色谱分析结果显示,苯妥英钠组和硝苯地平组药物释放良好,长期保持较高的浓度,环孢素组药物释放浓度下降较快,不能保持长期较高浓度；3.在SD雄性大鼠上颌磨牙区制备的牙龈退缩模型成功,实验组牙龈相比于对照组出现了退缩；4.苯妥英钠和硝苯地平的PLGA缓释微球对大鼠牙槽骨骨质的再生和牙龈胶原的增加有促进作用,环孢素没有表现出促进作用。其中,苯妥英钠的药效最为显著,新生牙槽骨面积、新生牙槽骨高度显著高于对照组(p0.01),牙龈厚度显著高于对照组(p0.01)。结论：1.苯妥英钠局部缓释给药可以显著促进牙周组织再生,可以用于治疗牙龈退缩和牙槽骨吸收。2.PLGA微球作为一种生物相容性良好的药物载体,它缓释的特性、微小的尺寸,可能是最适宜用于治疗牙周组织疾病的载体,起到了组织支架的作用,应用前景较为广阔。
[Abstract]:The experimental objective: to establish an animal model of drug induced gingival hyperplasia in SD male rats. Experimental methods: two representative drugs for gingival hyperplasia were selected as antiepileptic drugs, phenytoin sodium and calcium channel blocker nifedipine respectively. The time of administration was set for 25 days, 50 days and 75 days. The rats were divided into groups according to the time of administration. The rats were given the drug experiment. The rats were given one time daily. After the time of administration, the rats were killed and the maxillary and periodontal tissues were collected. The changes of gingival thickness were observed and measured under the stereoscopic microscope. Then paraffin embedded sections. HE staining and immunohistochemistry, and analysis of the results of the experiment. Experimental results: under the long-term oral administration of phenytoin or nifedipine, the gums of the SD male rats showed a more obvious proliferation (P0.01 or P0.05). Compared with the control group, the thickness of the gingiva in the experimental group was thickened and the gingival epithelium was elongated. Conclusion: phenytoin and nifedipine are long. The effect of local injection on gingival tissue of rats. The effect of local injection of second partial phenytoin on gingival of rats: a local injection of phenytoin solution to gingival hyperplasia in the gingiva of the maxillary molar of SD male rats, to observe the effect of phenytoin on the gingival of SD rats. Method: SD male rats were anaesthetized with the method of self control. A solution of phenytoin sodium was injected under the buccal gully of the maxillary first molar on one side of the maxillary rat. The dose was given once a day and the dose was 20ul. The rats were sacrificed for 24 hours and 40 days later. The rats' maxillary and periodontal tissues were collected and the gums were observed and measured. Changes in thickness. Experimental results: the results of the collection of analysis data after the end of the administration showed that after local injection of phenytoin sodium solution, the gums of the experimental group did not proliferate (P0.05) compared with the control group. Conclusion: local injection of phenytoin solution can not induce gingival hyperplasia in rats. It is necessary to improve the drug prescription. The effects of phenytoin sodium, nifedipine and cyclosporin's Poly (PLGA) microspheres on periodontal tissue regeneration: phenytoin, nifedipine and cyclosporin are the three major drug induced gingival proliferative drugs (antiepileptic, calcium channel antagonists, immunosuppressants). Biocompatible material PLGA as a carrier, the three drugs were introduced into the gingival regression model of rats respectively, and the treatment effect of three drugs on gingival retraction was tested by the way of sustained release drug delivery. The experimental method: 1. preparation of PLGA sustained-release microspheres of phenytoin, nifedipine, cyclosporin and 2. by HPLC. The drug components, release concentration and release time of three kinds of PLGA sustained-release microspheres were released in vitro; 3. the animal model of gingival contraction was prepared in the maxillary molar area of SD male rats; 4. the three kinds of drug loaded microspheres were introduced into the alveolar bone defect of the gingival retraction rats. After 2 months, the material was sacrificed and the gingival hyperplasia was observed under the stereoscopic microscope. Microscopically CT observation of alveolar bone regeneration and paraffin embedded section of maxillary and periodontal tissues of rats were used to detect the expression of collagen fibrin related proteins, osteoblasts and osteoclast related proteins. Experimental results: 1. PLGA sustained-release microspheres of three kinds of drugs were successfully prepared, and the morphology and scanning electron microscope were good in the literature and 2. high efficiency. The results of liquid chromatography analysis showed that the drug release in the phenytoin group and nifedipine group was good and maintained a high concentration for a long time. The drug release concentration in the cyclosporin group decreased rapidly and could not maintain a high concentration for a long time. 3. the gingival retraction model of the maxillary molar area of SD male rats was successfully developed, and the gingival of the experimental group appeared compared with the control group. PLGA sustained-release microspheres of 4. phenytoin and nifedipine promoted the regeneration of alveolar bone and the increase of gingival collagen in rats, and cyclosporin did not promote the effect. Among them, phenytoin sodium was most effective. The area of new alveolar bone and the height of new alveolar bone were significantly higher than that of the control group (P0.01), and the thickness of the gums was significant. Higher than the control group (P0.01). Conclusion: 1. phenytoin sodium can significantly promote periodontal tissue regeneration, can be used to treat gingival contraction and alveolar bone absorption of.2.PLGA microspheres as a biocompatible drug carrier, its sustained release characteristics, small size, may be the most suitable for the treatment of periodontal tissue disease. The carrier has played an important role in tissue scaffold and has wide application prospects.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R781.4

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