电穿孔介导的基因治疗对兔下颌骨牵引区新生骨骨密度与强度的影响

Objective To explore the effect of electroporation mediated gene therapy on bone mineral density and strength of new-formed bone in mandibular distraction gap, so as to enhance the osteogenesis and shorten the distraction term. Methods New-Zeland rabbits were employed. The distraction began after 3 days of latency period at the rate of 0. 8 mm per day for 7 days. After distraction, the rabbits were randomly divided into 5 groups to receive injection in the distraction gap with recombinant plasmid 2 μg(0. 1μg/μl)pIRES-hVEGF165-hBMP2 in group A, with recombinant plasmid pIRES-hBMP2 in group B, with recombinant plasmid pIRES-hVEGF165 in group C, with pIRES in group D, and with normal saline (NS) in group E. After injection, electroporation was performed in all the groups. After 1 week, 2 weeks, 4 weeks and 8 weeks of consolidation, all the animals underwent X-ray and quantitative computed tomography (QCT). The new-formed bone in distraction gap was selected as regions of interest (ROI) to measure the bone mineral density( BMD). Then the rabbits were sacrificed and the new-formed bone samples were harvested to detect 3-point crushing strength. Results BMD of newly formed bone in group A, B and C was markedly higher than that in group D and E (P < 0. 01 ). After 2 weeks of consolidation, BMD in group A was much higher than that in the other groups, but there was no difference between group B and C. After 4 weeks of consolidation, BMD in group A and B was markedly higher than that in group C, D and E ( P < 0. 01 ). After 8 weeks of consolidation, BMD in group A was markedly higher than that in the other groups. While the BMD was not significantly different between group B and C, but the BMD in group B and C was higher than that in group D and E ( P < 0. 01 ). After 4 weeks of consolidation, the 3-point crushing strength of newly formed bone in group A was markedly higher than that in group B,C, D and E ( P < 0. 01 ) , which was still the same after 8 weeks of consolidation. And the crushing strength in group B was higher than that in group C, D and E ( P < 0.05 ). Conclusions Electroporation-mediated transfection of recombinant plasmid pIRES-hVEGF165-hBMP2 could greatly enhance osteogenesis and calcification. A combination of VECF and BMP may promote osteogenesis and angiogenesis simultaneously, so as to magnify the effect of each growth factor, resulting a synergetic effect.     

局部皮瓣在面部创伤性皮肤软组织缺损急诊修复中的应用

目的 探索依据整形外科原则应用局部皮瓣急诊修复面部创伤性软组织缺损的效果.方法 对32例面部皮肤软组织即时性创伤性缺损,于严格清创后,根据创面缺损情况,按整形外科原则分别设计皮下浅表肌腱膜系统(SMAS)蒂皮瓣、V-Y推进皮瓣、眼轮匝肌蒂岛状皮瓣、鼻唇沟皮瓣等修复创面,精细缝合关闭伤口.结果 32例伤口均Ⅰ期愈合,随访6～18个月,无明显瘢痕增生,无功能障碍,无需Ⅱ期修复,美容效果良好.结论 遵照整形外科基本原则采用局部皮瓣修复面部创伤性皮肤软组织缺损,可获得满意的效果,值得临床推广应用.     

电穿孔介导的基因治疗对兔下颌骨牵引区新生骨骨密度与强度的影响

目的 探索电穿孔介导的基因治疗对下颌骨DO过程中牵引间隙新生骨密度与强度的影响,从而为促进下颌骨DO新骨生成,缩短牵引周期,减少并发症提供新思路.方法 以新西兰大白兔为实验动物模型,于术后3 d开始下颌骨牵引,每天0.8 mm,连续牵引7 d后,将实验动物分为5组.A组:在牵引区注射2 μg(O.1μg/μl)重组质粒pIRES-hVEGF165-hBMP2;B组:在牵引区注射2 μg(0.1μg/μl)重组质粒pIRES-hBMP2;C组:在牵引区注射2 μg(0.1μg/μl)重组质粒pIRES-hVEGF165;D组:在牵引区注射2 μg(0.1μg/μl)空质粒pIRES;E组:在牵引区注射相同剂量的生理盐水.5组实验动物均施加电穿孔刺激.各组分别于固定期第1、2、4、8周行X线及QCT检查.选整个牵张间隙新生骨痂部分为兴趣区,测定骨密度.然后处死动物.取材测量牵引区新生骨的三点抗压强度.结果 A、B、C组新生骨痂密度各时相点新生骨痂密度明显高于D、E组(P<0.01).固定2周,A组明显高于各组,但B、c组间比较差异无统计学意义.固定4周,A、B组明显高于C、D、E组(P<0.01).固定8周A组明显高于B、c、D、E组(P<0.01).B、C组间比较差异无统计学意义,但高于D、E组(P<0.01).固定4周,A组新生骨的三点抗压强度明显高于B、C、D、E组(P<0.01).固定8周,A组仍明显高于各组(P<0.01),且B组也明显高于c、D、E组(P<0.05).结论 电脉冲介导的pIRES-hVEGF165-hBMP2重组质粒体内转染可使牵引区获得较满意的骨再生和骨化成熟进程,其新骨骨化、改建过程均超过对照组.提示联合应用BMP与VEGF,可能会实现成骨与血供的联合重建,并且使单一生长因子的效应放大,使骨愈合的速度加快.     

牵引成骨中细胞因子的表达及作用

牵引成骨技术（distractionosteogenesis,DO）因其能在原位快速成骨而被称为是一种内源性组织工程技术（endogenous tissues engineer）。自1992年McCarthy首次将DO技术用于下颌骨畸形患者的治疗以来,经过十多年的基础研究与临床实践,DO在颅颌面外科缺损修复重建中的临床运用越来越受到重视。     

电穿孔介导的基因治疗对兔下颌骨牵引区新生骨骨密度与强度的影响

Objective To explore the effect of electroporation mediated gene therapy on bone mineral density and strength of new-formed bone in mandibular distraction gap, so as to enhance the osteogenesis and shorten the distraction term. Methods New-Zeland rabbits were employed. The distraction began after 3 days of latency period at the rate of 0. 8 mm per day for 7 days. After distraction, the rabbits were randomly divided into 5 groups to receive injection in the distraction gap with recombinant plasmid 2 μg(0. 1μg/μl)pIRES-hVEGF165-hBMP2 in group A, with recombinant plasmid pIRES-hBMP2 in group B, with recombinant plasmid pIRES-hVEGF165 in group C, with pIRES in group D, and with normal saline (NS) in group E. After injection, electroporation was performed in all the groups. After 1 week, 2 weeks, 4 weeks and 8 weeks of consolidation, all the animals underwent X-ray and quantitative computed tomography (QCT). The new-formed bone in distraction gap was selected as regions of interest (ROI) to measure the bone mineral density( BMD). Then the rabbits were sacrificed and the new-formed bone samples were harvested to detect 3-point crushing strength. Results BMD of newly formed bone in group A, B and C was markedly higher than that in group D and E (P < 0. 01 ). After 2 weeks of consolidation, BMD in group A was much higher than that in the other groups, but there was no difference between group B and C. After 4 weeks of consolidation, BMD in group A and B was markedly higher than that in group C, D and E ( P < 0. 01 ). After 8 weeks of consolidation, BMD in group A was markedly higher than that in the other groups. While the BMD was not significantly different between group B and C, but the BMD in group B and C was higher than that in group D and E ( P < 0. 01 ). After 4 weeks of consolidation, the 3-point crushing strength of newly formed bone in group A was markedly higher than that in group B,C, D and E ( P < 0. 01 ) , which was still the same after 8 weeks of consolidation. And the crushing strength in group B was higher than that in group C, D and E ( P < 0.05 ). Conclusions Electroporation-mediated transfection of recombinant plasmid pIRES-hVEGF165-hBMP2 could greatly enhance osteogenesis and calcification. A combination of VECF and BMP may promote osteogenesis and angiogenesis simultaneously, so as to magnify the effect of each growth factor, resulting a synergetic effect.     

持续弹性外牵引对雌性小型猪乳头碱性成纤维细胞生长因子和β1转化生长因子表达的影响

目的观察持续弹性外牵引过程中雌性小型猪乳头及其支撑组织中碱性成纤维细胞生长因子（bFGF）和β1转化生长因子（TGF-β1）表达的变化,探讨持续弹性外牵引矫治乳头内陷的机制。方法选用3只雌性小型猪,每只各12个乳头;随机选取4个乳头作为空白对照组,其余乳头安置自制弹性可调式牵拉矫治器进行持续外牵引。于牵引后第2、4、8、12周分别各切取1个空白对照组乳头和2个被牵引乳头,行免疫组织化学染色,观察不同时期bFGF和TCF-β1的表达情况。结果牵引后各时期bFGF及TGF-β1的表达均较对照组明显增加,差异有统计学意义（P〈0．01）。bFGF的阳性表达在牵引4周时达到最高水平,与12周时相比差异有统计学意义（P〈0．05）。TOF-β1的阳性表达在牵引2周时达到最高水平,同8周、12周牵引组相比差异有统计学意义（P〈0．05）。bFGF和TGF-β1表达呈显著正相关。结论持续弹性外牵引可刺激小型猪乳头组织中bFGF和TGF-β1的合成和分泌增加。     

下颌骨牵引成骨过程中基因干预对牵引区转化生长因子β1表达的影响

背景:局部基因治疗能促进牵引区新骨的生成,但关于基因治疗后对局部生长因子表达的影响目前尚不清楚。目的:观察电穿孔介导的基因治疗对兔下颌骨牵引成骨过程中转化生长因子β1表达的影响。方法:新西兰大白兔双侧下颌骨截骨后3d开始下颌骨牵引,0.8mm/d,连续牵引7d后,随机分为5组,分别在牵引区注射2μg(0.1g/L)重组质粒pIRES-hVEGF165-hBMP2、pIRES-hBMP2、pIRES-hVEGF165、空质粒pIRES及相同剂量的生理盐水。之后施加电穿孔刺激。结果与结论:免疫组织化学染色发现转化生长因子β1主要在细胞胞浆中表达,给药7d时骨端骨细胞、编织骨痂骨细胞、骨痂表面成骨细胞呈转化生长因子β1染色阳性;14d时新生成的编织骨痂骨细胞、骨痂表面成骨细胞、肉芽组织中的间质细胞、单核巨细胞、多核巨细胞转化生长因子β1染色阳性;28d时转化生长因子β1阳性细胞明显减少。其中注射重组质粒pIRES-hVEGF165-hBMP2、pIRES-hBMP2、pIRES-hVEGF165后转化生长因子β1的表达明显多于注射空质粒pIRES及生理盐水(P<0.05或P<0.01)。说明基因治疗能促进转化生长因子β1的表达,促进牵引区细胞基质的形成和新骨生成。     

不同时期转染基因对牵引区新骨生成的影响

背景：前期研究表明,电穿孔介导的重组质粒pIRES-hBMP2-hVEGF165能促进牵引区早期新生血管的形成和新骨形成。 目的：观察不同时机转染基因对兔下颌骨牵张成骨过程中牵引区新骨生成的影响,探索基因导入的最佳转染时间,以获得更好的治疗效果。 方法：新西兰大白兔48只,全麻下行双侧下颌骨截骨及牵引器植入后,采用随机区组法分成4组,分别于造模后即刻、牵引开始时、牵引结束时在双侧牵引区注射2 μg(0.1 g/L)重组质粒pIRES-hBMP2-hVEGF165,3组均予电穿孔刺激;单纯牵引组单纯牵引不行基因转染。各组于造模后3 d开始以0.8 mm/d、1次/d的速率进行牵引,连续牵引10 d;各组分别于固定期1,2,4,8周处死3只兔子,切取下颌牵引区新生组织行组织学检测和形态计量学分析。 结果与结论：组织学检查和形态计量学分析发现,牵引期转染组与即刻转染组、固定期转染组、单纯牵引组比较间隙内有更多的新生血管、成骨细胞和间充质细胞等成分,各时点新生骨量与新生骨小梁宽度明显高于后者(P < 0.05)。表明在牵引开始时(牵引期)进行基因转染较其他时间转染促进新骨生成作用明显,能够获得最佳的促进新骨生成的效果,提示牵引期是下颌骨基因治疗的最佳时机。     

电穿孔介导的基因治疗对下颌骨牵引新骨生成影响的组织形态计量学研究

目的：探索电穿孔介导的基因治疗对下颌骨牵引成骨（distraction osteogenesis,DO）过程中牵引间隙新骨生成的影响。方法：采用新西兰大白兔50只,双侧下颌骨截骨安装牵引器后3天开始牵引,每天0.8mm,连续牵引7天后,将实验动物分为5组,每组10只：A组：在牵引区注射2μg（0.1μg/μl）pIRES-hVEGF165-hBMP2;B组：在牵引区注射2μg（0.1μg/μl）pIRES-hBMP2。C组：在牵引区注射2μg（0.1μg/μl）pIRES-hVEGF165;D组：在牵引区注射2μg（0.1μg/μl）空质粒pIRES;E组：在牵引区注射相同剂量的生理盐水。5组实验动物均施加电穿孔刺激。各组分别于固定期第2、4、8周取材行组织学检查和形态计量学分析,测定牵引区新生骨量和新生骨小梁宽度。结果：A、B、C组新生骨量及新生骨小梁宽度明显高于D、E组（P〈0.01）。且A组明显高于B、C组,但B、C组,D、E组间无显著性差异。结论：电脉冲介导的pIRES-hVEGF165-hBMP2重组质粒体内转染可能实现成骨与血供的联合重建,使单一生长因子的效应放大,促进牵引区新骨的生成。     

外鼻整形术在鼻翼退缩分型患者中的应用效果

目的 观察外鼻整形术在鼻翼退缩分型患者中的应用效果。方法 选取四川悦好医学美容医院美容 外科2021年1月-2023年1月收治的60例鼻翼退缩分型患者,按照随机数字表法分为对照组和观察组,各 30例。对照组行鼻孔缩小术,观察组行外鼻整形术,比较两组外鼻整形参数、鼻美学指标、临床疗效、鼻 通气功能指标、鼻外形畸形情况、鼻腔通气评分及并发症发生情况。结果 观察组鼻长、理想鼻长、额鼻 角、鼻唇角优于对照组,差异有统计学意义（P＜0.05）;观察组鼻尖突出度高于对照组,鼻孔暴露度低于 对照组,差异有统计学意义（P ＜0.05）;观察组治疗总有效率为96.67%,高于对照组的86.67%,差异有 统计学意义（P ＜0.05）;观察组鼻腔最小截面积、鼻腔容积、鼻腔呼气吸气阻力各项鼻通气功能指标优 于对照组,差异有统计学意义（P ＜0.05）;观察组鼻外形畸形评分、鼻腔通气评分低于对照组,差异有 统计学意义（P ＜0.05）;两组均未见并发症发生。结论 外鼻整形术在鼻孔暴露度鼻翼退缩分型患者中的 应用效果优于鼻孔缩小术,对改善患者鼻孔暴露度、鼻部美观度等具有积极作用,更具临床应用价值。