猕猴腭裂动物模型的建立及牵张成骨整复腭部组织缺损

发布时间：2018-03-10 11:03

本文选题：腭裂　切入点：牵张成骨　出处：《天津医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 目的腭裂是最常见的先天性发育畸形之一,其发病率在我国高达新生儿的2‰左右。腭部骨与软组织的缺损畸形,严重影响患儿口颌系统功能及精神心理健康。以往传统的整复手术都是通过转移邻近组织瓣关闭裂隙、后延软腭长度等途径,以恢复腭部的解剖形态和生理功能,但均不能整复腭部的骨质缺损,恢复错位的肌肉附着。加之术后大量瘢痕形成影响患儿的颜面部生长发育,往往出现继发性的上颌发育不足。给患者及其家庭带来影响深远的社会心理障碍。近年来利用牵张成骨(Distraction Osteogenesis,DO)技术矫治骨骼畸形和缺损成为新的研究热点。DO以其操作简便、创伤小、形成瘢痕少、无需植骨,在形成新骨的同时,附着于其表面肌肉等软组织也相应得到伸展等独特优点逐渐得到青睐。本研究探索利用灵长类动物猕猴为研究对象,建立与人类解剖结构非常接近的腭裂动物模型,观察该模型的制作可行性及其效果,为今后的临床治疗研究建立实验平台,并且尝试使用DO技术关闭裂隙,观察牵张区域在不同时间点新骨生成的变化发展规律。同时自行开发设计具有专利知识产权的新型口内纯钛腭裂口内牵张矫治器,并观察其在DO应用过程中的效能、稳定性及适用性。方法 1、采用25只亚成体猕猴为实验对象,性别随机,3-4岁龄,体重4.7-6.4kg。利用外科手术在动物一侧硬腭部切除一约25×6mm~2大小矩形软硬组织,形成口鼻腔相通的洞穿性缺损,建立腭裂动物模型。术后6周安乐处死2只动物X-ray检查及大体组织观察裂隙形态,从而验证和判断该模型是否建立成功。 2、在上述方法成功建立模型的基础上,以骨切开术形成骨运送盘,安装自行设计的牵张器,延迟期5天后开始以0.8mm×1次/天的牵张速率牵张5～7天结束,固定6周后拆除牵张器。分别在牵张结束后的第1、2、4、6、8、12、24周安乐处死动物3只,通过X-ray检查、扫描电镜观察及组织学切片染色观察等方面研究,观察各时间点新骨的不同形态特征及其变化规律。结果 1、所有动物安全度过手术,患侧鼻孔有食物溢出、偶尔呛咳等表现,与人类腭裂患者表现相似,裂隙未发生自发性愈合,成功建立猕猴腭裂动物模型。 2、自行研制的口内纯钛牵张器顺利完成牵引及固定过程,除一例1颗压紧螺帽脱落仍保持稳定外,均完好保存至拆除。可靠性、稳定性及其牵张效能均高。 3、利用DO成功关闭模型动物的腭部裂隙。 4、X线影像学检查可见不同时间点间隙新骨不断形成钙化至接近正常骨密度。 5、扫描电镜下,可见新骨由类骨质产生到逐渐钙化成骨的超微结构变化过程。 6、组织学观察牵张区域由中央到边缘呈现新生类骨质逐渐矿化成骨小梁直至成熟改建,未见软骨骨痂。结论 1、利用外科手术建立的猕猴腭裂模型,能够顺利适应手术及其牵张操作过程,是进行腭裂以及其他原因导致的腭部获得性组织缺损牵张整复研究的良好实验动物模型。 2、本实验的纯钛腭裂口内牵张器使用方便,固位良好,能够顺利完成牵张及固定操作过程,效果稳定。 3、本研究0.8mm×1次/天的牵张速率及6周的固定期成骨效果良好。 4、大体组织学观察、X-ray检查及扫描电镜的观测的牵张区域的改变证实新骨的形成方式为膜内成骨。
[Abstract]:The purpose of cleft palate is one of the most common congenital malformation, the incidence rate in our country up to around 2. The neonatal palatal defect of bone and soft tissue deformity, seriously affect the patients' function of the stomatognathic system and mental health. The traditional surgery is through the transfer of the adjacent flap closed fracture after. The extension of the length of the soft palate and other ways to restore the anatomical morphology and physiological function of palate, but can not repair the palatal bone and restore normal muscle attachments. In addition to a large number of postoperative scar formation in infants with facial growth, often appear secondary maxillary hypoplasia to patients and their families to bring social and psychological obstacles far-reaching.
In recent years the use of distraction osteogenesis (Distraction Osteogenesis DO) technique for the treatment of skeletal deformities and defects become a hot research topic in the new.DO for its simple operation, small trauma, less scar formation, without bone graft in the formation of new bone at the same time, attached to the surface of muscle and soft tissue has the unique advantages of extension has been favored. This study explores the use of primate animal monkeys as the research object, the establishment of animal model of cleft palate structure is very close to the human anatomy, observe the production feasibility and effectiveness of the model, establish the experimental platform for the study of clinical treatment in the future, and try to use DO technology to close the cleft, observe the development of the traction law Zhang at the different time change the new bone formation. At the same time to develop new export with proprietary intellectual property in pure titanium palatal distraction appliance in the fissure, and observe the application effect in DO Yes, stability and applicability.
Method
1, the 25 sub adult rhesus monkeys as the experimental object, random sex, age 3-4, body weight 4.7-6.4kg. by surgery in the animal side of the hard palate resection of a 25 * 6mm~2 rectangular soft tissue, the formation of nasal cavity defects, the establishment of animal model of cleft palate. After 6 Zhou Anle killed 2 animal X-ray check and general observation of fracture morphology, thus verifying and judging whether the model was successfully established.
2, the model was successfully established based on the above method, the osteotomy was formed the carrier plate, installed distrator designed, delayed 5 days after the date of starting with 0.8mm * 1 / day stretch rate of stretch over 5~7 days, 6 weeks after fixation removed distractor in distraction respectively. After the end of the week 1,2,4,6,8,12,24 rats were euthanized by animal 3, X-ray examination, observation etc. histologic observation and scanning electron microscope observation of different forms of organization, characteristics and change rules of the new bone at each time point.
Result
1, all animals passed the operation safely. There were food spills and occasional choking cough on the affected nostrils. Similar to those of human cleft palate patients, there was no spontaneous healing of fissures. A rhesus monkey cleft palate animal model was successfully established.
2, the self-developed intraoral titanium distractor successfully completed the traction and fixation process. Except for one case of 1 compression nuts, the stability of the screw cap remained stable, and all of them were well preserved until the dismantling. The reliability, stability and distraction efficiency were all high.
3, the clefts of the palate of the model animals were successfully closed by DO.
4, X-ray imaging showed that the new bone formed in different time points formed calcification to close to the normal bone density.
5, under scanning electron microscope, the ultrastructural changes of the new bone from osteoid to gradual calcification and osteogenesis can be seen.
6, the histological observation of the distraction area from the central to the edge showed that the new bone like bone gradually mineralized into the bone trabecula straight to the mature reconstruction, and no cartilaginous callus was found.
conclusion
1, the rhesus monkey cleft palate model established by surgery can adapt to operation and stretching process smoothly. It is a good experimental animal model for palatine and other palatal acquired tissue defect distraction.
2, the pure titanium palate retainer in this experiment is easy to use and has good retention, and can successfully complete the process of traction and fixation, and the effect is stable.
3, the results of this study were good at 0.8mm x 1 / day and 6 weeks in the fixation period.
4, gross histological observation, X-ray examination and changes in the distraction area observed by scanning electron microscopy confirmed that the formation of the new bone was the osteogenesis in the membrane.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R782.2;R-332

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