CAJ | 학술논문

目的探讨电穿孔介导的基因治疗对下领骨牵引成骨过程中早期血管生成的影响.方法 32只新西兰大白兔随机分为4组:质粒+电穿孔组(A组),质粒组(B组),生理盐水+电穿孔组(C组),空白对照组(D组).各组动物分别于注射后1、3、7、14 d处死,取牵引区组织进行组织学检查、电镜观察、CD34免疫组织化学染色及微血管密度检测.结果 A、B组血管内皮细胞呈增殖活跃状态;C、D组多数血管内皮细胞部分呈现退变及凋亡早期改变.免疫组化染色发现,转染后第1天血管壁内皮细胞浆CD34表达较弱;第3、7、14天,牵引区肉芽组织血管内皮细胞均出现CD34阳性表达.A组CD34阳性表达较B组强,A、B组的CD34表达持续阳性且呈上升趋势;C、D组表达最弱,CD34阳性表达维持在第1天水平上平稳波动.结论电穿孔介导的pIRES-hVEGF165-EGFP重组质粒体内转染能够促进牵引区早期微血管的生成,使局部血管增生、渗入,增加骨断端的血流量.对调节和促进骨的生长和修复过程具有重要作用.
목적 탐토전천공개도적기인치료대하령골견인성골과정중조기혈관생성적영향.방법 32지신서란대백토수궤분위4조:질립+전천공조(A조),질립조(B조),생리염수+전천공조(C조),공백대조조(D조).각조동물분별우주사후1、3、7、14 d처사,취견인구조직진행조직학검사、전경관찰、CD34면역조직화학염색급미혈관밀도검측.결과 A、B조혈관내피세포정증식활약상태;C、D조다수혈관내피세포부분정현퇴변급조망조기개변.면역조화염색발현,전염후제1천혈관벽내피세포장CD34표체교약;제3、7、14천,견인구육아조직혈관내피세포균출현CD34양성표체.A조CD34양성표체교B조강,A、B조적CD34표체지속양성차정상승추세;C、D조표체최약,CD34양성표체유지재제1천수평상평은파동.결론 전천공개도적pIRES-hVEGF165-EGFP중조질립체내전염능구촉진견인구조기미혈관적생성,사국부혈관증생、삼입,증가골단단적혈류량.대조절화촉진골적생장화수복과정구유중요작용.
Objective To explore the effect of electroporation mediated gene therapy on angiogene-sis of the distraction area during early mandibular distraction osteogenesis (DO). Methods Thirty-two New-Zeland rabbits were randomly divided into 4 groups: group A: recombinant plasmid pIRES-VEGF165-EGFP and electroporation; group B: recombinant plasmid pIRES-VEGF165-EGFP; group C: normal saline (NS) and electroporation and group D: control group. The rabbits were sacrificed at 1d, 3d, 7d and 14d after injection, respectively. The distraction area tissue was removed for histological examination and electron microscopy, and immunohistochemical stain for CD34 was performed to detect the microvessel density. Results Generation of vascular endothelial cells (VEC) in the group A and group B were active, and majority of VEC in groups C and D took on early change of cataplasia and apoptosis. The immunohistochemical stain for CD34 showed that it expressed weakly at the first day after transfection, and at 3,7,14 days after transfection, CD34 of VECs in the distraction area expressed positively. CD34 expression was the strongest in group A (P<0. 05), and there was significant difference among three groups and different time, respectively.Compared to each other, CD34 of VECs expressed positively with a tendency to rise in the groups A and B. But it fluctuated at the level of the expression at the first day in the groups C and D. Conclusion Electroporation-mediated transfecting recombinant plasmid could promote angiogenesis during early stage of mandibular DO. It could promote local vascular proliferation and penetration, increase the blood flow of broken ends in fractured bone. It indicates that electroporation-mediated transfecting recombinant plasmid play an important role in regulating and promoting growth and reparative process of the bone.