小鼠牙胚在腎包膜及口腔頰黏膜下成牙能力的對(duì)比研究
發(fā)布時(shí)間：2018-08-31 14:15
【摘要】：背景及目的 近年來(lái)，牙齒組織工程以及基于牙齒發(fā)育和干細(xì)胞理論的牙齒再生技術(shù)，逐漸成為口腔醫(yī)學(xué)的研究熱點(diǎn)，，同時(shí)，建立一種合適的器官培養(yǎng)模式也是該領(lǐng)域的重點(diǎn)問(wèn)題。本實(shí)驗(yàn)旨在對(duì)比分析腎包膜和口腔黏膜環(huán)境對(duì)小鼠牙胚成牙的適宜度，為牙齒再生技術(shù)探尋較為理想的培養(yǎng)環(huán)境。 方法 選擇健康成年ICR小鼠50只，其中雌鼠30只，雄鼠20只，體重約20g。本實(shí)驗(yàn)共分為2組：1.實(shí)驗(yàn)組：實(shí)驗(yàn)組A-腎包膜下移植（選取右腎作為移植部位，挑開(kāi)腎囊膜后隨機(jī)選取牙胚植入），共3只小鼠，每只鼠移植12個(gè)牙胚，共36個(gè)牙胚；實(shí)驗(yàn)組B-口腔黏膜下移植（分別于小鼠的左右側(cè)下頜切牙旁頰黏膜處作切口，隨機(jī)選取牙胚植入），共18只小鼠，每只鼠移植2個(gè)牙胚，共36個(gè)牙胚；2.正常對(duì)照組：實(shí)驗(yàn)組C-取1~2只懷孕14.5天的孕鼠，不做任何處理，讓其自然生產(chǎn)。于術(shù)后3周、4周、8周三個(gè)時(shí)間點(diǎn)處死實(shí)驗(yàn)動(dòng)物后獲取標(biāo)本，進(jìn)行大體標(biāo)本觀察、制作組織切片、HE染色、硬度測(cè)試、拉曼光譜分析等。 結(jié)果 所有動(dòng)物術(shù)后均存活良好，腎包膜及口腔黏膜下均能檢測(cè)到移植物。牙胚在腎包膜及口腔黏膜下的發(fā)育情況如下所述。 1.大體形態(tài)學(xué)觀察結(jié)果顯示：腎包膜下組牙冠形態(tài)與同時(shí)期正常鼠磨牙形態(tài)相一致，尖窩發(fā)育明顯；而口腔黏膜下組在各時(shí)間點(diǎn)牙冠體積均小于腎包膜下組，且牙齒尖窩發(fā)育狀態(tài)不如腎包膜下組明顯。在兩個(gè)實(shí)驗(yàn)組中均無(wú)牙根生成。 2.組織學(xué)染色結(jié)果顯示：各組HE染色均可見(jiàn)正常的牙體組織結(jié)構(gòu)，在兩個(gè)實(shí)驗(yàn)組中可見(jiàn)上皮根鞘的組織結(jié)構(gòu)但未進(jìn)一步延伸；和腎包膜下組3~4周相比，在黏膜下形成的牙釉質(zhì)層和牙本質(zhì)層較薄，成釉細(xì)胞及成牙本質(zhì)細(xì)胞分化不明顯；兩個(gè)實(shí)驗(yàn)組8周時(shí)，均出現(xiàn)成釉細(xì)胞及成牙本質(zhì)細(xì)胞退化，牙髓內(nèi)細(xì)胞減少，部分形成機(jī)化組織，呈現(xiàn)纖維組織交織成網(wǎng)狀。 3.成牙概率統(tǒng)計(jì)結(jié)果顯示：同一移植時(shí)間點(diǎn)比較，腎包膜下組移植的成牙率均高于口腔黏膜下移植組，而在同一組內(nèi)，隨著移植周期的增長(zhǎng)，成牙率有所下降。 4.移植物硬度和彈性模量結(jié)果顯示：在釉質(zhì)的硬度比較上，僅口腔黏膜4周組的釉質(zhì)硬度比成年鼠牙低，差異具有統(tǒng)計(jì)學(xué)意義（P＜0.05），而其他組間兩兩比較的差別均無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05）。在釉質(zhì)的彈性模量比較上，腎包膜下3周后的移植物釉質(zhì)彈性模量比成年鼠牙略低，但差別無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05）；腎包膜下4周和口腔黏膜下4周后的移植物釉質(zhì)彈性模量均低于成年鼠牙組和腎包膜3周的釉質(zhì)彈性模量，差異具有顯著的統(tǒng)計(jì)學(xué)意義（P＜0.01），但是這兩組之間的差異無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05）。各組間牙本質(zhì)的硬度及彈性模量無(wú)顯著性差異（P＞0.05）。 5.拉曼光譜分析結(jié)果顯示：各組的拉曼光譜趨勢(shì)基本一致，在961cm-1處有最大峰值，但是在2947cm-1處，口腔黏膜3周組有明顯的峰值，遠(yuǎn)高于其他各組。 結(jié)論 1.對(duì)于ED14.5的牙胚來(lái)說(shuō)，相對(duì)口腔黏膜環(huán)境而言，腎包膜下環(huán)境培育3~4周更能發(fā)育成相對(duì)完整的牙齒。 2.口腔黏膜環(huán)境所形成牙齒的諸多性狀與腎包膜環(huán)境下發(fā)育的牙齒相比雖有差異，但其對(duì)牙胚的生長(zhǎng)發(fā)育還是具有一定的影響。
[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.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：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.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R783
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