Ctsk基因敲除调控H型血管参与小鼠牙槽窝愈合的研究
Study of Ctsk knockout regulating type H blood vessel formation and tooth extraction socket healing in mice
张武阳 1李登科 2王一名 1李元 1陈禹喆 1郑雪妮 1周宏志 1胡开进 1薛洋1
作者信息
- 1. 710032 西安,口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病临床医学研究中心,空军军医大学第三附属医院口腔颌面外科
- 2. 南部战区空军医院口腔科
- 折叠
摘要
目的:观察组织蛋白酶K(CTSK)对小鼠牙槽窝愈合及H型血管生成的作用.方法:利用CRISPR/Cas9技术构建Ctsk基因敲除小鼠,通过基因测序、大体观、Micro-CT及免疫组化对基因敲除情况进行鉴定;随后选择8周龄野生(wide type,WT)和Ctsk基因敲除小鼠拔除左侧上颌第一磨牙,于术后第7、10、14、21、28和35天处死各组小鼠(n=3),利用体视显微镜观察牙槽窝软组织愈合情况,利用Micro-CT观察骨组织愈合情况,利用免疫荧光染色观察牙槽窝愈合过程H型血管生成情况.结果:Ctsk基因敲除不影响小鼠牙槽窝黏膜软组织愈合,但能显著促进牙槽窝骨质愈合,且Ctsk基因敲除小鼠牙槽窝内H型血管生成显著增多.结论:Ctsk基因敲除能够增强H型血管生成,进而促进小鼠牙槽窝骨质愈合.
Abstract
Objective:To study the effects of cathepsin K(CTSK)on the healing process of tooth extraction socket and type H blood vessel angiogenesis in mice.Methods:Ctsk knockout(Ctsk-/-)mice were generated by CRISPR/Cas9 technology,and genotype sequen-cing,general observation,Micro-CT and immunohistochemistry were performed to confirm successful knockout of Ctsk.Then 8 week-old WT and Ctsk-/-mice were used to establish the tooth extraction modle by extracting the left maxillary first molars,and the mice were sac-rificed at the day 7,10,14,21,28 and 35 respectively(n=3)after tooth extraction.Then samples were subjected to stereo microscope and Micro-CT examination.Immunofluorescence staining was used to study the effect of Ctsk knockout on type H blood vessel angiogene-sis.Results:Ctsk knockout did not affect the soft tissue healing of tooth extraction socket,but significantly promoted the bone healing process,and Ctsk deficency significantly enhanced type H blood vessel angiogenesis in the tooth extraction socket.Conclusion:Ctsk knockout can enhance type H vessel angiogenesis,and promote bone healing process of tooth extraction socket in mice.
关键词
组织蛋白酶K/拔牙创愈合/H型血管/骨再生Key words
Cathepsin K/Healing of tooth extraction/Type H blood vessel/Bone regeneration引用本文复制引用
基金项目
军事口腔医学国家重点实验室自主研究课题(2019ZA09)
陕西省重点研发计划(2019SF-173)
出版年
2024