VEGF抗体靶向血管治疗增生性瘢痕的研究

目的探讨VEGF抗体靶向血管治疗对烧伤后增生性瘢痕的治疗作用。方法将人的增生性瘢痕植入裸鼠肩胛部皮下,建立裸鼠增生性瘢痕移植模型。3周后,用不同剂量血管内皮生长因子单克隆抗体进行瘢痕内注射治疗。治疗3周后解剖取材,通过测量瘢痕的大小观察瘢痕的体积变化,用Masson染色和免疫组化技术观察移植瘢痕胶原和微血管数量的变化。结果与PBS对照组相比,靶向血管治疗3周后,增生性瘢痕的体积明显减小,瘢痕胶原和微血管明显减少。结论VEGF抗体靶向血管治疗具有抑制增生性瘢痕微血管生成、胶原合成及抑制瘢痕生长的作用。     

VEGF抗体靶向血管治疗增生性瘢痕的研究

目的探讨VEGF抗体靶向血管治疗对烧伤后增生性瘢痕的治疗作用.方法将人的增生性瘢痕植入裸鼠肩胛部皮下,建立裸鼠增生性瘢痕移植模型.3周后,用不同剂量血管内皮生长因子单克隆抗体进行瘢痕内注射治疗.治疗3周后解剖取材,通过测量瘢痕的大小观察瘢痕的体积变化,用Masson 染色和免疫组化技术观察移植瘢痕胶原和微血管数量的变化.结果与PBS对照组相比,靶向血管治疗3周后, 增生性瘢痕的体积明显减小, 瘢痕胶原和微血管明显减少.结论 VEGF抗体靶向血管治疗具有抑制增生性瘢痕微血管生成、胶原合成及抑制瘢痕生长的作用.     

血管内皮生长因子抗体靶向血管治疗对增生性瘢痕Ⅰ型胶原蛋白表达的影响

目的观察血管内皮生长因子(VEGF)抗体靶向血管治疗对人增生性瘢痕Ⅰ型胶原蛋白在裸鼠体内表达的影响。方法将1％TBSA深Ⅱ度创面愈合后的增生性瘢痕组织块(取自1例女性烧伤患者)植入48只BALA／C裸鼠肩胛部皮下,建立裸鼠增生性瘢痕移植模型。术后3周,将裸鼠分为大剂量组、中剂量组、小剂量组及对照组,每组12只,分别用0．01 mol／L灭菌磷酸盐缓冲液(PBS)稀释的15、10、5μg／ml VEGF单克隆抗体200μl以及等量、同浓度的PBS进行瘢痕内直接注射,每周2次,持续3周。术后45 d,测量各组裸鼠瘢痕组织的大小,计算体积;以HE染色行组织学观察;采用逆转录聚合酶链反应与蛋白质印迹法分析瘢痕组织Ⅰ型前胶原蛋白mRNA和Ⅰ型胶原蛋白的表达。结果大剂量组、中剂量组、小剂量组瘢痕体积分别为(55．3±4．1)、(67．9±5．7)、(78．9±5．5)mm3;与对照组(85．0±7．3)mm3比较,大剂量组、中剂量组瘢痕体积明显变小(P< 0．05)。大剂量组、中剂量组血管和成纤维细胞较少,胶原纤维减少,排列较整齐。与对照组比较,大剂量组和中剂量组Ⅰ型前胶原蛋白mRNA和Ⅰ型胶原蛋白表达明显降低;小剂量组与之接近。结论VEGF抗体靶向血管治疗可抑制增生性瘢痕血管形成、胶原表达及瘢痕生长。     

其他

血管内皮生长因子抗体靶向血管治疗对增生性瘢痕Ⅰ型胶原蛋白表达的影响,颏下岛状皮瓣一期修复下咽肿瘤切除后非环周缺损,瘢痕研究,[编者按]     

以血管内皮生长因子受体为靶点抑制瘢痕血管增生的研究

目的　以血管内皮生长因子受体 2 (VEGFR 2 /KDR)为靶点,观察其抗体对烧伤增生性瘢痕新生血管形成的作用。方法　以人烧伤增生性瘢痕移植于裸鼠建立动物模型。治疗组KDR多抗分为10mg/L和5mg/L两组,以磷酸盐缓冲液(PBS)组和空白对照组作对照,每周2次,治疗1、2、3周分别观察瘢痕体积、组织形态学、微血管定量及Ⅰ、Ⅲ型胶原含量的变化。结果　治疗3周后的瘢痕体积(mm3 )、血管密度(个/mm2 )、Ⅰ、Ⅲ型胶原含量(阳性面积) ,KDR两治疗组均与对照组差异有统计学意义(P <0 .0 5 )。形态学显示瘢痕组织内有大量的血管内皮细胞坏死和血管闭塞,成纤维细胞凋亡,对照组变化不明显。结论　KDR抗体可通过抑制血管的形成治疗增生性瘢痕     

增生性瘢痕中血管内皮细胞生物学功能的研究

目的 探讨增生性瘢痕中血管内皮细胞的生物学功能及其与瘢痕形成的关系。方法 取人增生性瘢痕组织和正常皮肤组织，进行组织学观察。分离和纯化2种标本中的血管内皮细胞，应用酶联免疫吸附测定法分别检测单个血管内皮细胞中转化生长因子β1，（TGF-β1）、成纤维细胞生长因子2（FGF2）、血小板源性生长因子（PDGF）、内皮素1（ET-1）和血管内皮生长因子（VEGF）的水平。结果光学显微镜下可见正常皮肤微血管数目较少；增生性瘢痕微血管数目增多，血管狭长扭曲甚至闭塞。透射电镜可见增生性瘢痕中毛细血管管腔狭窄，有内皮细胞脱落。增生性瘢痕中血管内皮细胞分泌TGF-β1、PDGF、ET-1、VEGF、FGF2的水平分别为（60±8）、（30±4）、（0．12±0．03）、（52±5）、（18．1±1．2）μg／个细胞，明显低于正常皮肤（P〈0．05）。结论 增生性瘢痕中血管内皮细胞生物学功能减退，可能与瘢痕中胶原的大量产生和缺氧有关。     

VEGF和TSP-1在病理性瘢痕中的表达及其意义

目的 研究血管内皮生长因子(VEGF)和TSP-1(thrombospondin 1)在病理性瘢痕中的表达及其临床意义.方法 应用免疫组化SP法检测正常皮肤、扁平瘢痕、增生性德痕和瘢痕疙瘩组织中VEGF、TSP-1蛋白的表达并进行统计学分析.结果 病理性瘢痕组织中VEGF蛋白表达增高,与正常皮肤、扁平瘢痕对照组比较差异有统计学意义(P<0.05);病理性瘢痕组织中TSP-1蛋白表达显著低于正常皮肤、扁平瘢痕对照组(P<0.05);四类组织中VEGF蛋白和TSP-1蛋白呈负相关(P<0.05).结论 VEGF和TSP-1的表达与病理性瘢痕形成机制有关,VEGF可能通过诱导血管生成而促进瘢痕增生,TSP-1降低导致病理性瘢痕中血管的增生,并可能抑制VEGF的升高,从而导致病理性瘢痕的形成.     

血管内皮生长因子与病理性瘢痕

各类创伤创面愈合后常形成病理性瘢痕,随之发生外形改变、引发畸形及功能障碍,给患者带来巨大的心理和生理损害[1].病理性瘢痕包括增生性瘢痕(hypertrophic scar)和瘢痕疙瘩(keloid),是皮肤损伤后肉芽组织过度增生修复的结果.创伤修复需经历炎性反应、肉芽组织形成及组织重塑等主要阶段,每一阶段都有血管及其相关生物活性物质的参与,新生血管对于创面愈合起着至关重要的作用,为快速生长的细胞提供氧气和营养支持,以促进创伤组织的修复.对于增生性瘢痕和瘢痕疙瘩的组织学研究已经发现,其中有大量新生血管的存在,成纤维细胞合成大量的胶原可能与瘢痕组织内大量异常新生的血管有关.血管内皮生长因子(vas-cular endothelial growth factor,VEGF)与血管生成密切相关,研究也发现,VEGF在增生性瘢痕和瘢痕疙瘩中高度表达[2-4],因此,VEGF及其受体已经成为预防和治疗瘢痕增生研究的热门靶点.     

重组血管生成抑制剂Ad-METH-1对兔耳增生性瘢痕的抑制

目的 将已经成功构建、携带有METH-1基因的腺病毒表达载体pAdEasy-meth1,转染增生性瘢痕动物模型,研究血管靶向基因治疗对增生性瘢痕的抑制作用.方法 复制兔耳增生性瘢痕模型,于创面上皮化后10天,兔耳瘢痕局部注射携带有目的基因METH-1的重组腺病毒颗粒,30天后,用激光多普勒血流仪观察兔耳瘢痕微循环的变化,标本取材,行HE染色、CD34染色、核仁区嗜银颗粒染色,研究分析血管靶向基因治疗对兔耳瘢痕组织血管生成、微循环血流灌注及成纤维细胞增殖的影响.结果 与对照组相比,血管靶向治疗30天后,兔耳瘢痕微血管生成、微循环血流灌注及成纤维细胞增殖受到明显抑制.结论 在瘢痕形成早期,应用Ad-METH-1进行血管靶向治疗,对兔耳增生性瘢痕的形成,有明显的抑制作用.     

重组血管生成抑制剂Ad-METH-1对兔耳增生性瘢痕的抑制

目的将已经成功构建、携带有METH-1基因的腺病毒表达载体pAdEasy-meth1,转染增生性瘢痕动物模型,研究血管靶向基因治疗对增生性瘢痕的抑制作用.方法复制兔耳增生性瘢痕模型,于创面上皮化后10天,兔耳瘢痕局部注射携带有目的基因METH-1的重组腺病毒颗粒,30天后,用激光多普勒血流仪观察兔耳瘢痕微循环的变化,标本取材,行HE染色、CD34染色、核仁区嗜银颗粒染色,研究分析血管靶向基因治疗对兔耳瘢痕组织血管生成、微循环血流灌注及成纤维细胞增殖的影响.结果与对照组相比,血管靶向治疗30天后,兔耳瘢痕微血管生成、微循环血流灌注及成纤维细胞增殖受到明显抑制.结论在瘢痕形成早期,应用Ad-METH-1进行血管靶向治疗,对兔耳增生性瘢痕的形成,有明显的抑制作用.     

烧伤后增生性瘢痕患者不同时期血清中PⅢ NP的浓度变化及其意义

目的 探讨烧伤后增生性瘢痕患者不同时期血清中Ⅲ型前胶原氨基端肽(PⅢNP)浓度变化及其临床意义.方法 选择2007年8月-2009年8月收治住院的烧伤后增生性瘢痕患者共74例,临床上分属增生性瘢痕各个时期,根据瘢痕增生时段分组,应用放免法检测患者血清中PⅢNP的浓度变化,并比较PⅢNP的血清含量与瘢痕增生时段之间的关系.结果 烧伤后增生性瘢痕患者血清中PⅢNP含最和瘢痕增生时段关系密切,早期瘢痕增生患者血清中PⅢNP含量开始增加,随着增生性瘢痕的发展,其血清含量进一步的增加,到4～6个月的增生性瘢痕中,PⅢNP含量达到了高峰.伴随瘢痕的成熟,PⅢNP浓度又逐渐下降.结论 烧伤后增生性患者血清中PⅢNP的含量可较好地反映增生性瘢痕组织中的胶原代谢活跃程度,是增生性瘢痕增生活跃程度较为敏感的标志物,可以用做临床来指导临床瘢痕治疗.     

The effect of the angiogenesis inhibitor, angiostatin, on hypertrophic scarring

Introduction: Hypertrophic scarring is commonly seen by plastic surgeons in China. Since the etiology of hypertrophic scarring is still unknown, the only reliable treatment is surgical excision. To understand if angiogenesis plays an important role in the formation of hypertrophic scars, we investigated the effect of angiostatin, a potent angiogenesis inhibitor, on hypertrophic scar formation. Our hypothesis is that angiogenesis is required for increased scar formation and angiogenesis inhibition may be one of the methods that can be used to prevent the formation of hypertrophic scars. Methods: We have developed a reliable model in rabbits that results in hypertrophic scarring by creating a 6mm x 6mm full thickness skin wound on both ears. The cDNA for angiostatin is cloned into the pcDNA 3.1 mammalian expression vector. After the wounds re-epithelialized but prior to excessive scarring, the angiostatin expression vector was injected with Lipofectin 2000 once every two days. The expression of angiostatin was confirmed by RT-PCR. The scar tissue was harvested 14 days after injection and processed for histology and total protein. Histology was examined with routine stains, the amount of collagen deposition in the scar tissue was detected by proline assay, and TGF-β1 and VEGF expression was detected by western blot. Results: Compared to the control injection scar, the injection of angiostatin led to a much more normal-looking of scar in the rabbit ear. The proline assay demonstrated that the injection of the angiostatin expression vector resulted in much less collagen in the scar tissue. Western blot analysis showed there was less TGF-β1 and VEGF protein expression in the treated ear compared to the control. Conclusion: The introduction of a vector over-expression angiostatin can result in the decreased formation of hypertrophic scars in a rabbit ear model. This is corroborated by evidence of decreased collagen deposition, the primary extracellular matrix component of scars. In addition, we demonstrate the decreased expression of τηε pro-fibrosis growth factor, TGF-1, and the potent angiogenic factor, VEGF. These data suggest that angiogenesis inhibitors may have a potential role in the treatment of hypertrophic scarring.     

烧伤后增生性瘢痕患者不同时期血清中PⅢNP的浓度变化及其意义

目的探讨烧伤后增生性瘢痕患者不同时期血清中Ⅲ型前胶原氨基端肽（PⅢNP）浓度变化及其临床意义。方法选择2007年8月-2009年8月收治住院的烧伤后增生性瘢痕患者共74例,临床上分属增生性瘢痕各个时期,根据瘢痕增生时段分组,应用放免法检测患者血清中PⅢNP的浓度变化,并比较PⅢNP的血清含量与瘢痕增生时段之间的关系。结果烧伤后增生性瘢痕患者血清中PⅢNP含量和瘢痕增生时段关系密切,早期瘢痕增生患者血清中PⅢNP含量开始增加,随着增生性瘢痕的发展,其血清含量进一步的增加,到4～6个月的增生性瘢痕中,PⅢNP含量达到了高峰。伴随瘢痕的成熟,PⅢNP浓度又逐渐下降。结论烧伤后增生性患者血清中PⅢNP的含量可较好地反映增生性瘢痕组织中的胶原代谢活跃程度,是增生性瘢痕增生活跃程度较为敏感的标志物,可以用做临床来指导临床瘢痕治疗。     

Hypertrophic scar fibroblasts accelerate collagen gel contraction

Excessive contraction of hypertrophic scar and subsequent contracture formation are a formidable problem after thermal injury. A comparison between fibroblasts from hypertrophic scar and normal skin was made with the use of fibroblast-populated collagen lattices as a measure of cellular generated contractile forces. Hypertrophic scar and normal skin fibroblasts were mixed with soluble tendon collagen and Dulbecco's modified Eagle medium supplemented with 10% serum, and contraction was measured by serial area measurements. Parallel experiments in the presence of transforming growth factor-beta or anti-transforming growth factor-beta antibody examined the role of this cytokine on lattice contraction. Transforming growth factor-beta activity was measured in an additional set of 10 biopsy specimens. Hypertrophic scar fibroblasts contract lattices at a significantly faster rate than do normal skin fibroblasts. Exogenous transforming growth factor-beta increased lattice contraction by normal skin fibroblasts but had little effect on hypertrophic scar cell-populated lattices. The addition of anti-transforming growth factor-beta antibody decreased lattice contraction by both cell types. Transforming growth factor-beta activity was significantly increased in the hypertrophic scar biopsy specimens. Excessive scar contraction and post-burn scar contracture result from increased contraction forces generated by hypertrophic scar cells. This increased contractility appears to be mediated by increased endogenous presence of transforming growth factor-beta.     

Changes in VEGF and nitric oxide after deep dermal injury in the female, red Duroc pig-further similarities between female, Duroc scar and human hypertrophic scar

Despite decades of research, our understanding of human hypertrophic scar is limited. A reliable animal model could significantly increase our understanding. We previously confirmed similarities between scarring in the female, red, Duroc pig and human hypertrophic scarring. The purpose of this study was to: (1) measure vascular endothelial growth factor (VEGF) and nitric oxide (NO) levels in wounds on the female Duroc; and (2) to compare the NO levels to those reported for human hypertrophic scar. Shallow and deep wounds were created on four female Durocs. VEGF levels were measured using ELISA and NO levels with the Griess reagent. VEGF and NO levels were increased in deep wounds at 10 days when compared to shallow wounds (p < 0.05). At 15 weeks, VEGF and NO levels had returned to the level of shallow wounds. At 21 weeks, VEGF and NO levels had declined below baseline levels in deep wounds and the NO levels were significantly lower (p < 0.01). We found that VEGF and NO exhibit two distinctly different temporal patterns in shallow and deep wounds on the female Durocs. Furthermore, NO is decreased in female, Duroc scar as it is in human, hypertrophic scar further validating the usefulness of the model.