Acidic Fibroblast Growth Factor: Evaluation of Topical Formulations in a Diabetic Mouse Wound Healing Model

The efficacy of topical formulations of acidic fibroblast growth factor (aFGF) in healing of full-thickness wounds has been studied in a diabetic db + /db+ mouse model. The effect of several formulation variables, dose, and application frequency was examined. It was found that wound healing in diabetic animals treated with aFGF or placebo was slower than in their nondiabetic littermates. The availability of aFGF from the viscous vehicle employed in this study (1% hydroxyethyl cellulose) was demonstrated in vitro using diffusion cells. The viscous formulation of aFGF was equally effective in wound healing as a nonviscous formulation in phosphate-buffered saline. A formulation containing heparin (necessary for full biological and conformational stability of aFGF) at a mass ratio of 3:1 to aFGF was more efficacious than formulations with lower heparin: aFGF ratios. Wounds treated with three doses of 3.0 µg/cm² aFGF healed faster than those treated with a single dose of 3.0 µg/cm² aFGF. Three applications of 3.0 or 0.6 µg/cm² aFGF were equally effective in accelerating wound healing.     

Topical Application of Keratinocyte Growth Factor Conjugated Gold Nanoparticles Accelerate Wound Healing

Keratinocyte growth factor (KGF) has been demonstrated to specifically stimulate the multiplication and migration of keratinocytes. However, due to rapid degradation, the results of topical application of growth factors on wounds are unsatisfactory. In this study, we cross-linked KGF to the surface of gold nanoparticles (GNPs) and explored their effects on wound healing. The as-synthesized nanocomposite (KGF-GNPs) displayed good colloidal stability, decent biocompatibility as well as negligible cellular cytotoxicity. The in vitro cellular experimental results demonstrated that KGF-GNPs could effectively promote the proliferation of keratinocytes in contrast to bare GNPs or KGF. Furthermore, in animal full-thickness wound model, KGF-GNPs are more conducive to wound healing than bare GNPs or KGF. KGF-GNPs enhanced wound healing by promoting wound re-epithelialization rather than granulation. The superior biocompatibility, colloidal depressiveness and biological activity of this nanocomposite indicate that it could be utilized as a promising wound healing drug for clinical application in the future.     

miR-9a通过促进角质形成细胞增殖和迁移促进皮肤伤口愈合

目的 探讨miR-9a对大鼠皮肤伤口愈合的影响及其作用途径。方法 构建大鼠伤口愈合模型,检测miR-9a在不同时间点皮肤伤口愈合组织中的表达水平。过表达miR-9a,利用原位杂交试验、HE染色、免疫荧光染色、划痕试验和伤口愈合试验分析差异。结果 与损伤前相比,损伤后7 d皮肤组织中miR-9a表达显著升高（P<0.001）;过表达miR-9a组在损伤第14天和第21天损伤区域面积显著减小（P<0.05或P<0.001）。与对照组相比,转染miR-9a到角质形成细胞HaCaT后,EdU阳性细胞数量显著增加（P<0.001）、伤口愈合百分比显著升高（P<0.01）、迁移细胞的数量显著增加（P<0.001）。结论 miR-9a促进大鼠皮肤伤口愈合,其途径可能是通过促进角质形成细胞的增殖和迁移。     

碱性成纤维细胞因子促进创面修复的临床疗效

目的:观察碱性成纤维细胞因子(bFGF)对创面修复的疗效及毒副作用.方法:173例患者分为治疗组89例,其中急性创面55例,慢性难愈合创面34例,采用bFGF局部外用,每平方厘米150活性单位,qd,7 d为1个疗程;对照组84例,其中急性创面50例,慢性难愈合创面34例,除未用bFGF外,其他疗法同治疗组.结果:治疗组较对照组创面修复质量显著提高,创面愈合时间缩短,且无明显毒副作用.结论:bFGF具有促进创面修复作用,有很大临床应用价值.     

胶原蛋白海绵复合bFGF促进兔胫骨外露创面愈合的实验研究

目的研究胶原蛋白海绵复合bFGF对骨外露创面愈合的影响。方法 30只日本大耳白兔,随机分为A、B、C三组,每组10只。于胫骨上段前内侧作一1cm×1.5cm创面,暴露胫骨。A组创面喷洒bFGF后,胶原蛋白海绵覆盖,纱布包扎。B组覆盖胶原蛋白海绵扎。C组喷洒生理盐水。术后观察各组创面完全愈合的时间,测量并计算术后各时期创面收缩率。创面愈合后,取材,HE染色,显微镜下观察组织结构。结果大体观察:A、B、C组创面愈合时间分别为(25.12±1.46)d、(30.50±1.69)d、(33.38±1.77)d。A组较B、C组愈合时间提前,差异有统计学意义(P<0.01)。组织学观察:A组创面愈合质量优于B、C组。结论胶原蛋白海绵复合bFGF对兔胫骨外露创面愈合具有促进作用,提高创面愈合质量,减少瘢痕组织。     

bFGF对成纤维细胞增殖和胶原及纤维连接蛋白的影响

目的：探讨碱性成纤维细胞因子（ bFGF）调节皮肤创伤修复的作用机制。方法采用组织块贴壁法培养来源于正常皮肤和增生性瘢痕的成纤维细胞（FB）,加入含有不同浓度bFGF（0,0.1,1,10,100,1000 ng·mL-1）的无血清培养液培养72 h,采用细胞计数法和锥虫蓝染色检测各组FB增殖和凋亡,酶联免疫吸附法（ ELISA）和逆转录PCR （ RT-PCR）分别检测Ⅰ型、Ⅲ型胶原和纤维连接蛋白浓度及基因表达情况。结果 bFGF促进FB增殖,当浓度过高时则抑制FB增殖,促进其凋亡,并且增生性瘢痕FB增殖较来源于正常皮肤的FB缓慢;bFGF明显抑制增生性瘢痕FB Ⅰ型胶原产生,对来源于正常皮肤FB无影响;bFGF上调来源于正常皮肤FB的纤维连接蛋白基因表达,对增生性瘢痕FB中的表达无影响;bFGF对两种来源的FBⅢ型胶原的分泌和表达均无显著作用。结论 bFGF对来源正常皮肤和增生性瘢痕的FB具有不同的影响和作用机制,可能对创伤修复早期和瘢痕形成过程发挥作用。     

碱性成纤维生长因子与血管内皮生长因子联合对大鼠后肢动脉硬化闭塞血管再生作用机制的研究

目的探讨联合应用血管内皮生长因子(VEGF)与碱性成纤维细胞生长因子(bFGF)对大鼠后肢动脉硬化闭塞血管再生的作用机制。方法建立大鼠后肢动脉硬化闭塞模型60只,随机分为4组,生理盐水治疗组(NS组)15只,碱性成纤维细胞生长因子组(bFGF组)15只,血管内皮生长因子组(VEGF组)15只,血管内皮生长因子+碱性成纤维细胞生长因子(VEGF+bFGF组)15只。NS组、VEGF组分别隔日一次腹腔注射生理盐水1mL、100μg/LVEGF 1mL;bFGF组隔日一次后肢股内侧多点肌肉注射100μg/L bFGF 1mL;VEGF+bFGF组隔日一次腹腔注射100μg/L LVEGF 1mL+后肢股内侧多点肌肉注射100μg/L bFGF 1mL,共治疗21d。分别于7d、14d、21d观察大鼠后肢缺血情况如:间歇性跛行情况、皮肤颜色改变、皮温改变、足溃疡情况等。继续饲养至3个月后取后肢股内侧部位肌肉组织行免疫组化观察血管数。结果 VEGF+bFGF组微血管密度显著高于bFGF组、VEGF组及NS组,差异有显著性意义(P<0.05);其他各组间差异无显著性意义(P>0.05)。可见VEGF与bFGF联合应用可促进后肢动脉硬化闭塞大鼠的血管再生。结论 VEGF联合bFGF能促进大鼠后肢动脉硬化闭塞血管再生,为临床治疗下肢动脉硬化闭塞提供新的治疗依据。     

重组人酸性成纤维细胞生长因子治疗肛瘘术后创面的随机对照试验

目的：观察重组人酸性成纤维细胞生长因子（rh-aFGF）对肛瘘术后的创面促愈合作用.方法：237例肛瘘术后患者随机分为2组,观察组（n=125）给予rh-aFGF,对照组（n=112）给予重组人碱性成纤维细胞生长因子（bFGF）进行创面治疗.观察治疗3周后两组剩余创面大小、愈合率、创面祛腐时间、创面平均愈合时间、创面愈合率、肉芽生长开始时间等情况,及治疗后第7、14、21天肉芽组织生长、创面疼痛情况.结果：治疗3周后,观察组在减少创面面积,增强创面愈合率,缩短愈合时间、祛腐时间,以及肉芽组织开始生长时间等方面均优于对照组（P＜0.05或0.01）;治疗后第7天两组肉芽组织生长情况及疼痛情况差异无统计学意义（P＞0.05）,治疗第14天及第21天,两组肉芽组织生长及疼痛差异均有统计学意义（P＜0.05）.结论：rh-aFGF对肛瘘术后的促修复作用较bFGF更强,有效促进创面缩小、缩短祛腐时间与愈合时间、缩短肉芽生长开始时间、提高创面愈合率、有效提高肉芽组织生长、缓解创面疼痛等方面的作用.     

重组人酸性成纤维细胞生长因子促进儿童创面愈合的多中心、开放性随机对照试验

目的:观察重组人酸性成纤维细胞生长因子(rh-a FGF)对儿童手术或意外创面愈合的促进作用。方法:206例患儿随机分作2组,观察组给予rh-a FGF、对照组给予重组人碱性成纤维细胞生长因子(b FGF),分别进行创面治疗,观察治疗7 d后两组伤口愈合、患儿感染、脂肪液化等疗效情况,及治疗6个月后的瘢痕、色素沉着及不良反应等情况。结果:rh-a FGF、b FGF治疗7 d的愈合率分别为88.4%、74.2%,剩余创面面积分别为(2.2±1.1)cm2、(3.9±2.4)cm2;6个月后rh-a FGF组与b FGF组疤痕发生率分别为27.2%、67.5%,色素沉着发生率分别为30.1%、69.9%,两组比较差异均有统计学意义(P<0.05)。结论:rh-a FGF对儿童手术或意外创伤创面的促修复作用较b FGF更强,可有效促进创面愈合、缩短愈合时间、降低疤痕发生率及色素沉着发生率、有效减少疤痕面积及色素沉着面积。     

Improvement in Wound Healing by Epidermal Growth Factor (EGF) Ointment. I. Effect of Nafamostat,Gabexate, or Gelatin on Stabilization and Efficacy of EGF

The healing effect of human epidermal growth factor (hEGF) on open wounds was studied in rats. No improvement in wound healing was found by topical application of EGF alone to open wound sites. We found an ointment containing EGF and a protease inhibitor, nafamostat mesilate or gabexate mesilate, or gelatin accelerated the healing rate of open wounds. Significant increases in the dry weight of the wound site granulation tissue, uronic acid (as an index of acid mucopolysaccharide) and hydroxyproline (as an index of collagen) were observed by treatment with EGF ointment containing nafamostat compared with the controls. The effects of the protease inhibitor on wound healing were dose dependent. Nafamostat was more efficient than gabexate or gelatin on wound healing. The degradation of ¹²⁵I-EGF in wound tissue homogenate was significantly decreased in the presence of a protease inhibitor, such as nafamostat or gabexate, or gelatin. These findings indicate that the stabilization of EGF at the wound site is an important factor in permitting the expression of its healing effects and suggest that the ointment containing EGF and a stabilizing agent would be a suitable dosage form for acceleration of wound repair.     

碱性成纤维细胞生长因子对兔甲状软骨细胞的影响

目的 研究碱性成纤维细胞生长因子(bFGF)对免甲状软骨细胞的影响，探索软骨细胞体外扩增方法，为软骨组织工程提供理论依据。方法 分离鬼甲状软骨细胞，分4组培养，每组用含不同成分的培养基一第1组：只用DMEM培养基；第2组：DMEM培养基 10％小牛血清；第3组：DMEM培养基加bFGF(10ng/m1)；第4组：DMEM培养基 10％小牛血清 bFGF(10ng/m1)。采用软骨细胞计数观察各组细胞增殖情况。结果 在培养基成分相同时，补充bFGF组比未补充组细胞数目明显增多，即第3组多于第1组，第4组多于第2组。结论 bFGF能够促进单层培养中兔甲状软骨细胞的增殖，在软骨组织工程细胞增殖阶段，可能成为软骨细胞体外大量扩增的主要生长因子之一。     

模拟微重力环境对小鼠成纤维细胞增殖和创伤修复相关蛋白基因表达的影响

目的：研究微重力细胞培养系统( RCCS)模拟微重力环境下小鼠成纤维细胞( L929细胞)增殖及创伤修复相关蛋白基因表达的变化。方法将L929细胞随机分为微重力组和正常重力对照组,分别在RCCS中培养3、5、7天,流式细胞仪检测细胞周期,实时荧光定量聚合酶链反应检测创伤修复蛋白mRNA表达水平。培养第7天取两组细胞-微载体悬液观察细胞微丝结构形态变化。结果微重力组L929细胞的G1期细胞在第3、5、7天明显少于正常重力对照组(P<0．05);与正常重力对照组相比,微重力组S期细胞在第3、5天有所增加,第7天减少,G2/M期细胞在培养第3、7天升高,培养第5天下降(P<0．05)。培养第3天微重力组Ⅰ型胶原蛋白(ColⅠ)和Ⅲ型胶原蛋白(ColⅢ)、β转化生长因子( TGF-β) mRNA表达低于正常重力对照组( P<0．05);培养第5天微重力组Caspase-3 mRNA表达低于正常重力对照组,ColⅠ、TGF-β及Bcl-2 mRNA表达均高于正常重力对照组(P<0．05);培养第7天微重力组创伤修复相关蛋白mRNA表达均高于正常重力对照组( P<0．05)。模拟微重力环境下培养7 d对L929细胞微丝结构有影响但两组间差异无明显差别。结论模拟微重力环境能改变L929细胞周期转化、增强细胞增殖能力,并影响创伤修复相关蛋白的表达。     

反应停对人类肺腺癌细胞系血管内皮生长因子表达影响

目的：探讨反应停对亲代与耐顺铂人类肺腺癌细胞系A549与A549^DDP血管内皮生长因子（VEGF）表达的影响，及在克服肺腺癌顺铂耐药中的作用。方法：采用RT－PCR和免疫细胞化学方法，分别检测经反应停处理前后A549与A549^DDP VEGF mRNA与蛋白的表达水平。结果：生理剂量（6mg/L)反应停处理后第1－5d，A549 VEGF mRNA表达较处理前显著增加，A549^DDP VEGF mRNA表达较处理前显著降低。不同浓度反应停处理后第5d,A549gn A549^DDP VEGF mRNA表达水平存在显著性差异，且蛋白表达与其相应mRNA呈显著正相关。结论：生理剂量反应停显著上调A549 VEGF mRNA表达，但大剂量则抑制其表达，同时显著下调A549^DDP VEGF mRNA表达，呈剂量依赖性，VEGF蛋白表达与mRNA表达量一致。     

VEGF MVD在子宫腺肌病中的表达

目的 探讨子宫腺肌病异位内膜组织中血管内皮生长因子(VEGF)和微血管密度(MVD)的表达.方法 采用免疫组织化学方法检测子宫腺肌病异位内膜与正常子宫内膜中VEGF和MVD的表达.结果 子宫腺肌病异位内膜组织中VEGF和MVD在增生期和分泌期的表达均明显高于正常内膜(P＜0.05).结论 异位内膜组织中VEGF和MVD的高表达可能对子宫腺肌病的发生发展起重要作用.     

阿魏酸钠对类风湿关节炎患者血清VEGF和TNF-α表达的影响

目的:观察阿魏酸钠治疗类风湿关节炎(RA)的疗效及对血清血管内皮生长因子(VEGF)、肿瘤坏死因子-α(TNF-α)表达水平的影响。方法:43例RA患者随机分为治疗组与对照组,治疗组在对照组治疗基础上加用阿魏酸钠,连用4wk。ELISA法检测治疗前后血清VEGF、TNF-α水平。结果:治疗组与对照组总有效率分别为91.30%、75.00%(P<0.05),治疗后血清VEGF、TNF-α水平治疗组均显著低于对照组(P<0.05)。结论:阿魏酸钠能显著提高RA患者的疗效,并能影响血清VEGF、TNF-α水平。     

2型糖尿病患者血管内皮生长因子、C反应蛋白与动脉硬化的关系

目的研究2型糖尿病患者血管内皮生长因子（VEGF）和C反应蛋白（CRP）水平变化与动脉硬化的关系。方法酶联免疫分析法和免疫比浊法分别检测2型糖尿病患者（共63例,单纯糖尿病33例,糖尿病动脉硬化者30例）及健康人（31例）血浆VEGF、CRP水平,观察其水平变化与动脉硬化间的关系。结果糖尿病动脉硬化组血浆VEGF、CRP浓度明显高于单纯糖尿病组及正常对照组（P〈0．05）;Logistic回归分析示SBP、TC、TG、HDL、HbA．C、VEGF、CRP与动脉硬化的发生相关。多元相关分析示VEGF与WHR、SBP、FPG、TC、TG、HDL、HbAlc、CRP相关（P〈0．05）;CRP与WHR、SBP、FPG、TC、TG、HDL、HbA1c、VEGF相关（P〈0．05）。结论T2DM动脉硬化患者VEGF、CRP可能参与动脉粥样硬化形成过程。控制血压、血脂、血糖,减轻体质量,抗炎等综合治疗有益于延缓动脉硬化的发生与发展。     

VEGF及其受体与MMP-9在卵巢上皮性癌中的表达

目的：研究血管内皮生长因子（VEGF）及其受体、MMP-9的表达在卵巢上皮性癌侵袭转移方面的作用。方法：免疫组化SABC法检测VEGF及其受体、MMP-9蛋白的表达，并对VEGF、MMP-9蛋白表达及其相互关系进行秩和检验、Spearman等级分析，采用Kaplan-Meier分析进行生存分析。结果：VEGF广泛表达于卵巢上皮性癌的瘤细胞胞浆中，疾病晚期、分化差、腹水量多、残余瘤组织多的患者表达增强；VEGF受体表达于卵巢癌血管内皮细胞的胞浆．尤其是靠近癌巢处的内皮细胞；MMP-9表达于卵巢上皮性癌的间质细胞，疾病晚期、分化差的患者表达增强：VEGF与MMP-9在卵巢上皮性癌中的表达呈正相关。结论：VEGF与MMP-9在血管生长和局部侵袭及远处转移中可能有协同作用。     

非小细胞肺癌血清VEGF和PDGF测定的临床意义

目的：探讨血清血管内皮生长因子（VEGF）和血小板衍生生长因子（PDGF）测定在非小细胞肺癌诊断和预后判定中的意义。方法：采用双抗体夹心ABC—EHSA法测定31例非小细胞肺癌患者及30例健康对照组血清VEGF和PDGF的含量。结果：非小细胞肺癌患者血清VEGF和PDGF测定值均高于对照组（P〈0．01）。血清VEGF与非小细胞肺癌临床分期有关（P〈0.01），Ⅲ期和Ⅳ期非小细胞肺癌患者血清PDGF高于Ⅱ期患者（P〈0.01）。血清VEGF和PDGF测定值与非小细胞肺癌病理分型无关（P〉0．05）。非小细胞肺癌患者血清VEGF与PDGF测定值之间呈正相关（r=0．641，P〈0．01）。结论：检测血清VEGF和PDGF水平对非小细胞肺癌的诊断和预后判定具有一定价值。     

Quercetin Glucuronide Inhibits Cell Migration and Proliferation by Platelet-Derived Growth Factor in Vascular Smooth Muscle Cells

Many epidemiologic studies have reported that dietary flavonoids provide protection against cardiovascular disease. Quercetin, a member of the bioflavonoids family, has been proposed to have anti-inflammatory, anti-atherogenic, and anti-hypertensive properties leading to the beneficial effects against cardiovascular diseases. Recent studies demonstrated that orally administered quercetin appeared in plasma as glucuronide-conjugated forms in rats and humans. Therefore, we examined the effect of chemically synthesized quercetin glucuronide on platelet-derived growth factor (PDGF)-induced cell migration and kinase activation in cultured rat aortic smooth muscle cells (RASMCs). PDGF-induced RASMC migration was inhibited by quercetin 3-O-β-D-glucuronide (Q3GA). Q3GA also attenuated PDGF-induced cell proliferation in RASMCs. PDGF activated extracellular-signal regulated kinase (ERK) 1/2, c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein (MAP) kinase, and Akt in RASMCs. PDGF-induced JNK and Akt activations were suppressed by Q3GA, whereas ERK1/2 and p38 MAP kinase activations were not affected. We also confirmed that PDGF-induced JNK and Akt activations were inhibited by antioxidants, N-acetylcysteine and diphenyleneiodonium chloride, in RASMCs. These findings suggest Q3GA would be an active metabolite of quercetin in plasma and may possess preventing effects for cardiovascular diseases relevant to vascular smooth muscle cell disorders.     

Effect of Molecular Size of PEGylated Recombinant Human Epidermal Growth Factor on the Biological Activity and Stability in Rat Wound Tissue

The purpose of this study was to investigate the effect of size of polyethylene glycol (PEG) conjugated to recombinant human epidermal growth factor (rhEGF) on its stability in skin wound tissue and in vitro biological activity to find the desirable conjugate as topical therapeutic agent for wound healing. Site-specific PEGylation at N-terminus of rhEGF was performed with monomethoxy PEG-Butyraldehyde derivatives (MW 2, 5, and 20 kDa). Mono-PEG-rhEGFs retained 60–70% of biological activity of native rhEGF, and the effect of PEG size was not significant. The improvement of stability in the rat skin wound tissue was dependent on the increase of the PEG size attached. The degradation half-lives of native rhEGF, mono-PEG-2K-, ?5K-, and ?20K-rhEGFs were 1.1, 3.1, 5.2, and 41.5 hr, respectively. Therefore, mono-PEG-20K-rhEGF was considered to be the most desirable in terms of the increase of stability and the preservation of biological activity. This study suggests that the high molecular weight PEG at N-terminus of rhEGF would give a satisfactory stabilizing effect and thus may improve therapeutic efficacy in clinical use.