本文選題：SD大鼠 + 苯妥英鈉��； 參考：《武漢大學(xué)》2015年博士論文

【摘要】：第一部分 大鼠藥物性牙齦增生模型的建立實(shí)驗(yàn)?zāi)康模阂許D雄性大鼠為實(shí)驗(yàn)對(duì)象,建立藥物性牙齦增生的動(dòng)物模型。實(shí)驗(yàn)方法：選擇兩種致藥物性牙齦增生的代表藥物,分別為抗癲癇藥苯妥英鈉和鈣通道阻滯劑硝苯地平,給藥方式為全身給藥(口服),給藥時(shí)間設(shè)定為25天、50天、75天,按不同給藥時(shí)間對(duì)大鼠進(jìn)行分組,對(duì)大鼠進(jìn)行給藥實(shí)驗(yàn),每日給藥一次,給藥期結(jié)束后,處死實(shí)驗(yàn)鼠,收集上下頜骨及牙周組織,在體視顯微鏡下觀察并測(cè)量牙齦厚度的變化,然后石蠟包埋切片,HE染色及免疫組化,并分析實(shí)驗(yàn)結(jié)果。實(shí)驗(yàn)結(jié)果：在苯妥英鈉或硝苯地平長(zhǎng)期口服給藥下,SD雄性大鼠的牙齦表現(xiàn)出較為明顯的增生(p0.01或p0.05),與對(duì)照組比較發(fā)現(xiàn),實(shí)驗(yàn)組牙齦厚度增厚,牙齦上皮釘突伸長(zhǎng)。結(jié)論：苯妥英鈉和硝苯地平長(zhǎng)期全身用藥,可以誘導(dǎo)大鼠牙齦組織的藥物性增生。第二部分苯妥英鈉局部注射給藥對(duì)大鼠牙齦的作用實(shí)驗(yàn)?zāi)康模涸赟D雄性大鼠上頜磨牙的牙齦局部注射致牙齦增生的藥物苯妥英鈉溶液,觀察苯妥英鈉對(duì)SD大鼠牙齦的作用情況。實(shí)驗(yàn)方法：SD雄性大鼠行進(jìn)行全身麻醉,采用自身對(duì)照的方法,在大鼠上頜一側(cè)的第一磨牙頰側(cè)齦頰溝黏膜下注射苯妥英鈉溶液,每日給藥一次,給藥劑量為20ul,間隔24小時(shí),40天后處死實(shí)驗(yàn)鼠,收集大鼠上頜骨及牙周組織,觀察并測(cè)量大鼠牙齦厚度的變化。實(shí)驗(yàn)結(jié)果：給藥結(jié)束后收集分析數(shù)據(jù)結(jié)果顯示,局部注射苯妥英鈉溶液后,與對(duì)照組相比,大鼠實(shí)驗(yàn)組的牙齦并未出現(xiàn)增生(p0.05)。結(jié)論：以苯妥英鈉溶液的形式進(jìn)行局部注射給藥不能誘導(dǎo)大鼠牙齦增生,有必要改進(jìn)給藥方法和劑型。第三部分載苯妥英鈉、硝苯地平、環(huán)孢素的聚乳酸羥基乙酸(PLGA)緩釋微球?qū)ρ乐芙M織再生的作用實(shí)驗(yàn)?zāi)康模罕酵子⑩c、硝苯地平、環(huán)孢素是三大類致藥物性牙齦增生藥(抗癲癇藥,鈣通道拮抗劑,免疫抑制劑)的代表藥,通過一種生物相容性材料PLGA作為載體,將這三種藥物分別導(dǎo)入大鼠牙齦退縮模型中,以緩釋給藥的方式,檢驗(yàn)三種藥物,對(duì)牙齦退縮的治療效果。實(shí)驗(yàn)方法：1.制備苯妥英鈉、硝苯地平、環(huán)孢素三種藥物的PLGA緩釋微球；2.通過高效液相色譜法檢驗(yàn)三種PLGA緩釋微球在體外釋放的藥物成分、釋放濃度及釋放時(shí)間；3.在SD雄性大鼠上頜磨牙區(qū)制備牙齦退縮的動(dòng)物模型；4.將三種載藥微球?qū)胫苽浜玫难例l退縮大鼠牙槽骨缺損處,2個(gè)月后,處死取材,在體視顯微鏡下觀察牙齦增生情況,顯微CT觀察牙槽骨再生,大鼠上頜頜骨及牙周組織石蠟包埋切片,檢測(cè)膠原纖維相關(guān)蛋白,成骨細(xì)胞、破骨細(xì)胞相關(guān)蛋白的表達(dá)。實(shí)驗(yàn)結(jié)果：1.三種藥物的PLGA緩釋微球制備成功,宏觀形態(tài)及掃描電鏡下形態(tài)良好,符合文獻(xiàn)描述；2.高效液相色譜分析結(jié)果顯示,苯妥英鈉組和硝苯地平組藥物釋放良好,長(zhǎng)期保持較高的濃度,環(huán)孢素組藥物釋放濃度下降較快,不能保持長(zhǎng)期較高濃度；3.在SD雄性大鼠上頜磨牙區(qū)制備的牙齦退縮模型成功,實(shí)驗(yàn)組牙齦相比于對(duì)照組出現(xiàn)了退縮；4.苯妥英鈉和硝苯地平的PLGA緩釋微球?qū)Υ笫笱啦酃枪琴|(zhì)的再生和牙齦膠原的增加有促進(jìn)作用,環(huán)孢素沒有表現(xiàn)出促進(jìn)作用。其中,苯妥英鈉的藥效最為顯著,新生牙槽骨面積、新生牙槽骨高度顯著高于對(duì)照組(p0.01),牙齦厚度顯著高于對(duì)照組(p0.01)。結(jié)論：1.苯妥英鈉局部緩釋給藥可以顯著促進(jìn)牙周組織再生,可以用于治療牙齦退縮和牙槽骨吸收。2.PLGA微球作為一種生物相容性良好的藥物載體,它緩釋的特性、微小的尺寸,可能是最適宜用于治療牙周組織疾病的載體,起到了組織支架的作用,應(yīng)用前景較為廣闊。
[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.
【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R781.4

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本文編號(hào)：2049946