小鼠牙胚在肾包膜及口腔颊黏膜下成牙能力的对比研究
发布时间：2018-08-31 14:15
【摘要】：背景及目的 近年来，牙齿组织工程以及基于牙齿发育和干细胞理论的牙齿再生技术，逐渐成为口腔医学的研究热点，，同时，建立一种合适的器官培养模式也是该领域的重点问题。本实验旨在对比分析肾包膜和口腔黏膜环境对小鼠牙胚成牙的适宜度，为牙齿再生技术探寻较为理想的培养环境。 方法 选择健康成年ICR小鼠50只，其中雌鼠30只，雄鼠20只，体重约20g。本实验共分为2组：1.实验组：实验组A-肾包膜下移植（选取右肾作为移植部位，挑开肾囊膜后随机选取牙胚植入），共3只小鼠，每只鼠移植12个牙胚，共36个牙胚；实验组B-口腔黏膜下移植（分别于小鼠的左右侧下颌切牙旁颊黏膜处作切口，随机选取牙胚植入），共18只小鼠，每只鼠移植2个牙胚，共36个牙胚；2.正常对照组：实验组C-取1~2只怀孕14.5天的孕鼠，不做任何处理，让其自然生产。于术后3周、4周、8周三个时间点处死实验动物后获取标本，进行大体标本观察、制作组织切片、HE染色、硬度测试、拉曼光谱分析等。 结果 所有动物术后均存活良好，肾包膜及口腔黏膜下均能检测到移植物。牙胚在肾包膜及口腔黏膜下的发育情况如下所述。 1.大体形态学观察结果显示：肾包膜下组牙冠形态与同时期正常鼠磨牙形态相一致，尖窝发育明显；而口腔黏膜下组在各时间点牙冠体积均小于肾包膜下组，且牙齿尖窝发育状态不如肾包膜下组明显。在两个实验组中均无牙根生成。 2.组织学染色结果显示：各组HE染色均可见正常的牙体组织结构，在两个实验组中可见上皮根鞘的组织结构但未进一步延伸；和肾包膜下组3~4周相比，在黏膜下形成的牙釉质层和牙本质层较薄，成釉细胞及成牙本质细胞分化不明显；两个实验组8周时，均出现成釉细胞及成牙本质细胞退化，牙髓内细胞减少，部分形成机化组织，呈现纤维组织交织成网状。 3.成牙概率统计结果显示：同一移植时间点比较，肾包膜下组移植的成牙率均高于口腔黏膜下移植组，而在同一组内，随着移植周期的增长，成牙率有所下降。 4.移植物硬度和弹性模量结果显示：在釉质的硬度比较上，仅口腔黏膜4周组的釉质硬度比成年鼠牙低，差异具有统计学意义（P＜0.05），而其他组间两两比较的差别均无统计学意义（P＞0.05）。在釉质的弹性模量比较上，肾包膜下3周后的移植物釉质弹性模量比成年鼠牙略低，但差别无统计学意义（P＞0.05）；肾包膜下4周和口腔黏膜下4周后的移植物釉质弹性模量均低于成年鼠牙组和肾包膜3周的釉质弹性模量，差异具有显著的统计学意义（P＜0.01），但是这两组之间的差异无统计学意义（P＞0.05）。各组间牙本质的硬度及弹性模量无显著性差异（P＞0.05）。 5.拉曼光谱分析结果显示：各组的拉曼光谱趋势基本一致，在961cm-1处有最大峰值，但是在2947cm-1处，口腔黏膜3周组有明显的峰值，远高于其他各组。 结论 1.对于ED14.5的牙胚来说，相对口腔黏膜环境而言，肾包膜下环境培育3~4周更能发育成相对完整的牙齿。 2.口腔黏膜环境所形成牙齿的诸多性状与肾包膜环境下发育的牙齿相比虽有差异，但其对牙胚的生长发育还是具有一定的影响。
[Abstract]:Background and purpose
In recent years, dental tissue engineering and tooth regeneration based on tooth development and stem cell theory have gradually become a hot topic in stomatology. At the same time, the establishment of an appropriate organ culture model is also a key issue in this field. To explore an ideal culture environment for tooth regeneration technology.
Method
Fifty healthy adult ICR mice were selected, including 30 females and 20 males, weighing about 20 g. The experiment was divided into two groups: 1. Experimental group: A-kidney subcapsular transplantation (right kidney was selected as the transplantation site, and then the renal capsule was picked out and randomly selected to implant dental germs), a total of 3 mice, 12 dental germs per mouse, a total of 36 dental germs; Experimental group B-oral mucus. Submembranous transplantation (left and right mandibular incisor buccal mucosa incision, randomly selected dental germs implantation), a total of 18 mice, each mouse transplanted 2 dental germs, a total of 36 dental germs; 2. Normal control group: experimental group C - Take 1 to 2 pregnant mice 14.5 days pregnant, without any treatment, let them produce naturally. 3 weeks, 4 weeks, 8 weeks after surgery for three times Samples were obtained after point-killing, gross observation, histological section, HE staining, hardness test, Raman spectroscopy and so on.
Result
All the animals survived well after the operation. Grafts were detected in the renal capsule and oral submucosa. The development of tooth germs in the renal capsule and oral submucosa is described below.
1. The results of gross morphological observation showed that the crown morphology of the subcapsular group was consistent with that of the normal mouse molars at the same time, and the cuspid fossa development was obvious. The crown volume of the subcapsular group was smaller than that of the subcapsular group at each time point, and the cuspid fossa development was not as obvious as that of the subcapsular group.
2. Histological staining showed that normal dental tissues were observed in all groups with HE staining, and the epithelial root sheath was observed in both groups without further extension. At the 8th week, ameloblasts and odontoblasts degenerated, the cells in the pulp decreased, and some of the tissues formed organic tissue, showing a network of fibrous tissue.
3. The statistical results of dental formation probability showed that the rate of subcapsular renal transplantation was higher than that of Oral Submucosal transplantation at the same time point, and the rate of dental formation decreased with the increase of transplantation period in the same group.
4. The results of the hardness and elastic modulus of the graft showed that the hardness of the enamel in the 4-week group was lower than that in the adult rats (P < 0.05), but there was no significant difference between the other groups (P > 0.05). The elastic modulus of allograft enamel was slightly lower than that of adult rat teeth, but there was no significant difference between the two groups (P > 0.05); the elastic modulus of allograft enamel was lower than that of adult rat teeth and renal capsule after 4 weeks of renal capsule and oral mucosa (P < 0.01), but there was no significant difference between the two groups (P < 0.01). There was no significant difference in hardness and elastic modulus between each group (P > 0.05) (P > 0.05).
5. The results of Raman spectroscopy analysis showed that the trend of Raman spectroscopy in each group was basically the same, with the largest peak at 961 cm-1, but at 2947 cm-1, there was a significant peak at 3 weeks of oral mucosa, much higher than other groups.
conclusion
1. For ED14.5 tooth germs, the development of relatively complete teeth in subcapsular environment is better than that in oral environment.
2. Although the characteristics of teeth formed by oral mucosal environment are different from those of teeth developed under renal envelope environment, they still have some effects on the growth and development of tooth germs.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R783
[Abstract]:Background and purpose
In recent years, dental tissue engineering and tooth regeneration based on tooth development and stem cell theory have gradually become a hot topic in stomatology. At the same time, the establishment of an appropriate organ culture model is also a key issue in this field. To explore an ideal culture environment for tooth regeneration technology.
Method
Fifty healthy adult ICR mice were selected, including 30 females and 20 males, weighing about 20 g. The experiment was divided into two groups: 1. Experimental group: A-kidney subcapsular transplantation (right kidney was selected as the transplantation site, and then the renal capsule was picked out and randomly selected to implant dental germs), a total of 3 mice, 12 dental germs per mouse, a total of 36 dental germs; Experimental group B-oral mucus. Submembranous transplantation (left and right mandibular incisor buccal mucosa incision, randomly selected dental germs implantation), a total of 18 mice, each mouse transplanted 2 dental germs, a total of 36 dental germs; 2. Normal control group: experimental group C - Take 1 to 2 pregnant mice 14.5 days pregnant, without any treatment, let them produce naturally. 3 weeks, 4 weeks, 8 weeks after surgery for three times Samples were obtained after point-killing, gross observation, histological section, HE staining, hardness test, Raman spectroscopy and so on.
Result
All the animals survived well after the operation. Grafts were detected in the renal capsule and oral submucosa. The development of tooth germs in the renal capsule and oral submucosa is described below.
1. The results of gross morphological observation showed that the crown morphology of the subcapsular group was consistent with that of the normal mouse molars at the same time, and the cuspid fossa development was obvious. The crown volume of the subcapsular group was smaller than that of the subcapsular group at each time point, and the cuspid fossa development was not as obvious as that of the subcapsular group.
2. Histological staining showed that normal dental tissues were observed in all groups with HE staining, and the epithelial root sheath was observed in both groups without further extension. At the 8th week, ameloblasts and odontoblasts degenerated, the cells in the pulp decreased, and some of the tissues formed organic tissue, showing a network of fibrous tissue.
3. The statistical results of dental formation probability showed that the rate of subcapsular renal transplantation was higher than that of Oral Submucosal transplantation at the same time point, and the rate of dental formation decreased with the increase of transplantation period in the same group.
4. The results of the hardness and elastic modulus of the graft showed that the hardness of the enamel in the 4-week group was lower than that in the adult rats (P < 0.05), but there was no significant difference between the other groups (P > 0.05). The elastic modulus of allograft enamel was slightly lower than that of adult rat teeth, but there was no significant difference between the two groups (P > 0.05); the elastic modulus of allograft enamel was lower than that of adult rat teeth and renal capsule after 4 weeks of renal capsule and oral mucosa (P < 0.01), but there was no significant difference between the two groups (P < 0.01). There was no significant difference in hardness and elastic modulus between each group (P > 0.05) (P > 0.05).
5. The results of Raman spectroscopy analysis showed that the trend of Raman spectroscopy in each group was basically the same, with the largest peak at 961 cm-1, but at 2947 cm-1, there was a significant peak at 3 weeks of oral mucosa, much higher than other groups.
conclusion
1. For ED14.5 tooth germs, the development of relatively complete teeth in subcapsular environment is better than that in oral environment.
2. Although the characteristics of teeth formed by oral mucosal environment are different from those of teeth developed under renal envelope environment, they still have some effects on the growth and development of tooth germs.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R783
【相似文献】
相关期刊论文 前10条
1 杨帆,肖明振,赵守亮;人牙胚成纤维细胞生长因子18的基因克隆和序列分析[J];牙体牙髓牙周病学杂志;2003年05期
2 江宏兵;牙胚发生的位置和形态调控[J];国外医学.口腔医学分册;2003年04期
3 李锋,吴补领,余擎;小鼠牙胚体外发育过程中组织形态学变化的观察[J];中国临床康复;2004年14期
4 贺慧霞;金岩;史俊南;罗玉庆;常秀梅;;构建大鼠牙胚肾被膜下培养模型及其牙胚组织混合后的成牙能力[J];中国组织工程研究与临床康复;2007年02期
5 高玉光,韩建国,史俊南;维甲酸体外诱导鼠磨牙牙胚骨涎蛋白基因表达的研究[J];牙体牙髓牙周病学杂志;2000年01期
6 于金华,金岩,史俊南,聂鑫,王亦菁,庄Y
本文编号：2215240
本文链接：https://www.wllwen.com/yixuelunwen/kouq/2215240.html
最近更新
教材专著
