烧伤创面成纤维细胞周期、羟脯氨酸和胶原比例动态变化

目的　探讨深Ⅱ度烧伤创面愈合规律。　方法　通过 4 8只大鼠深Ⅱ度烧伤模型 ,动态观察烧伤后不同时间点创面成纤维细胞增殖周期、羟脯氨酸 (OHP)含量和Ⅰ /Ⅲ型胶原比值变化及病理组织学改变。　结果　伤后 10d真皮细胞s期细胞百分比达高峰 ,伤后 2 1d仍显著高于正常对照组。OHP含量伤后逐渐增加 ,伤后 14d达高峰 ,伤后 2 1d仍显著高于正常对照组。伤后Ⅰ /Ⅲ型胶原比值逐渐降低 ,并显著低于正常对照组 ,病理切片观察伤后 3～ 10d明显可见白细胞浸润带 ,伤后 14d可见上皮细胞自创缘及坏死组织界面爬行生长。　结论　深Ⅱ度烧伤创面伤后 3～ 10d以炎症反应为主。伤后 10～ 14d主要表现为成纤维细胞增殖和新生胶原合成 ,伤后 2 1d真皮的修复尚未结束。伤后 14～ 2 1d主要为表皮细胞增殖和迁移。     

烧伤创面羟脯氨酸含量动态变化的实验研究

为探讨浅Ⅱ度烧伤创面胶原含量的变化及其与创面上皮化的关系，我们对浅Ⅱ度烧伤创面的羟脯氨酸（ＯＨＰ）水平及创面的形态学变化作了动态检测。结果发现烧伤后早期创面表皮缺损，ＯＨＰ含量增高，伤后第３天达峰值，继之随着上皮化的完善，ＯＨＰ水平逐渐下降至正常。我们认为，浅Ⅱ度烧伤创面胶原的含量与上皮化程度有关，良好的上皮化过程有利于控制胶原的过度增生。表皮细胞源因子（ＥＤＦ）可能是表皮细胞调控抑制胶原合成的主     

烧伤创面羟脯氨酸含量动态变化的实验研究

为探讨浅Ⅱ度烧伤创面胶原含量的变化及其与创面上皮化的关系，我们对浅Ⅱ度烧伤创面的羟脯氨酸（ＯＨＰ）水平及创面的形态学变化作了动态检测。结果发现烧伤后早期创面表皮缺损，ＯＨＰ含量增高，伤后第３天达峰值，继之随着上皮化的完善，ＯＨＰ水平逐渐下降至正常。我们认为，浅Ⅱ度烧伤创面胶原的含量与上皮化程度有关，良好的上皮化过程有利于控制胶原的过度增生。表皮细胞源因子（ＥＤＦ）可能是表皮细胞调控抑制胶原合成的主要因素。     

精氨酸对烧伤创面愈合的影响

目的 探讨精氨酸对烧伤创面愈合的影响和量效关系。方法 通过大鼠深Ⅱ度烫伤模型,伤后肠道站充不同剂量精氨酸,观察烧伤创面愈合时间,测定烧伤创面愈合面积百分比,创面Ｉ型／Ⅲ型胶原比例,羟脯氨酸（ＯＨＰ）含量和真皮细胞增殖周期,结果 每天补充１００－４００ｍｇ／ｋｇ精氨酸可缩短烧伤创面愈合时间,增加烧伤创面中ＯＨＰ含量,降低烧伤创面Ｉ／Ⅲ型胶原比例,加速真皮细胞ＤＮＡ复制。比较喂不同剂量精氨酸各组,发现     

应用十二烷基硫酸钠—聚丙烯酰胺凝胶电泳分析兔Ⅰ型和…

应用十二烷基硫酸钠－聚丙烯酰胺凝胶电泳分析了兔皮中Ⅲ型胶原的含量。采用此法检测了正常兔皮伤后７２ｈ深Ⅱ度创面，伤后１月及１．５月的已愈创面组织中胶原成分的改变，旨在探讨深Ⅱ度烧伤早期及愈后皮肤胶原的变化，结果表明创面在伤后及愈合后组织中Ⅲ型胶原含量均显著减少，提示在创面愈合过程中，胶原修复的同时伴有Ⅲ型胶原的减少。     

类肝素对大鼠深Ⅱ度烫伤创面愈合的影响

目的探讨深Ⅱ度烧伤创面早期加深而延迟愈合的机理。方法采用大鼠深Ⅱ度烫伤模型，创面外用类肝素软膏，观察其对大鼠深Ⅱ度烫伤早期创面病理变化和创面愈合的影响，并测定了烧伤创面含水量，血浆和烧伤创面抗凝血酶Ⅲ（ＡＴ－Ⅲ）、纤维蛋白降解产物（ＦＤＰ）含量、烧伤创面羟脯氨酸含量、Ⅰ／Ⅲ型胶原比例、真皮细胞增殖周期及创面愈合时间。结果创面外用类肝素可以降低烧伤创面水肿程度，增加血浆ＡＴ－Ⅲ活性，增加烧伤创面中ＦＤＰ含量及羟脯氨酸含量，促使Ⅰ／Ⅲ型胶原比例降低，增加真皮细胞中ＤＮＡ合成，减轻烧伤创面早期加深，缩短大鼠深Ⅱ度烫伤创面愈合时间。结论类肝素具有促进大鼠深Ⅱ度烫伤创面愈合的作用。     

应用十二烷基硫酸钠—聚丙烯酰胺凝胶电泳分析兔Ⅰ型和Ⅲ型胶原的含量

应用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE 电泳)分析了兔皮中Ⅲ型胶原的含量。采用此法检测了正常兔皮伤后72h 深Ⅱ度创面、伤后1月及1.5月的已愈创面组织中胶原成分的改变,旨在探讨深Ⅱ度烧伤早期及愈后皮肤胶原的变化,结果表明创面在伤后及愈合后组织中Ⅲ型胶原含量均显著减少。提示在创面愈合过程中,胶原修复的同时伴有Ⅲ型胶原的减少。     

应用十二烷基硫酸钠－聚丙烯酰胺凝胶电泳分析兔Ⅰ型和Ⅲ型胶原的含量

应用十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ电泳）分析了兔皮中Ⅲ型胶原的含量。采用此法检测了正常兔皮伤后７２ｈ深Ⅱ度创面、伤后１月及１．５月的已愈创面组织中胶原成分的改变，旨在探讨深Ⅱ度烧伤早期及愈后皮肤胶原的变化，结果表明创面在伤后及愈合后组织中Ⅲ型胶原含量均显著减少。提示在创面愈合过程中，胶原修复的同时伴有Ⅲ型胶原的减少。     

类肝素对大鼠深Ⅱ度烫伤创面愈合的影响

目的探讨深Ⅱ度烧伤创面早期加深而延迟愈合的机理。方法采用大鼠深Ⅱ度烫伤模型,创面外用类肝素软膏,观察其对大鼠深Ⅱ度烫伤早期创面病理变化和创面愈合的影响,并测定了烧伤创面含水量,血浆和烧伤创面抗凝血酶Ⅲ(AT-Ⅲ)、纤维蛋白降解产物(FDP)含量、烧伤创面羟脯氨酸含量、Ⅰ/Ⅲ型胶原比例、真皮细胞增殖周期及创面愈合时间。结果创面外用类肝素可以降低烧伤创面水肿程度,增加血浆AT-重活性,增加烧伤创面中FDP含量及羟脯氨酸含量,促使Ⅰ/Ⅲ型胶原比例降低,增加真皮细胞中DNA合成,减轻烧伤创面早期加深,缩短大鼠深Ⅱ度烫伤创面愈合时间。结论类肝素具有促进大鼠深Ⅱ度烫伤创面愈合的作用。     

烧伤创面羟脯氨酸含量动态变化的实验研究

为探讨浅Ⅱ度烧伤创面胶原含量的变化及其与创面上皮化的关系,我们对浅Ⅱ度烧伤创面的羟脯氨酸(OHP)水平及创面的形态学变化作了动态检测。结果发现烧伤后早期创面表皮缺损,OHP含量增高,伤后第3天达峰值,继之随着上皮化的完善,OHP 水平逐渐下降至正常。我们认为,浅Ⅱ度烧伤创面胶原的含量与上皮化程度有关,良好的上皮化过程有利于控制胶原的过度增生。表皮细胞源因子(EDF)可能是表皮细胞调控抑制胶原合成的主要因素。     

表皮生长因子对大鼠深Ⅱ度烧伤创面愈合的影响

目的　进一步观察重组人表皮生长因子 (rhEGF)对深Ⅱ度烧伤创面愈合的促进作用。方法　采用大鼠深Ⅱ度烫伤模型 (以下称烧伤 ) ,创面分别外用rhEGF、肝素加rhEGF及等渗盐水。比较不同方式处理创面后的愈合时间 ,测定创面愈合率、创面含水量、羟脯氨酸 (OHP)含量及Ⅰ /Ⅲ型胶原比例 ,进行细胞DNA周期分析和组织学检查。　结果　外用rhEGF可使烧伤创面愈合时间缩短 2d,增加创面OHP含量 (P <0.0 5),降低Ⅰ /Ⅲ型胶原比例 (P <0.0 5),促进肉芽组织形成 ,加速细胞DNA复制 (P <0.0 5 )。加用肝素后可使创面愈合时间进一步缩短 2d,促进肉芽组织生长。伤后第 7天用药各组间比较 ,差异无显著性意义 (P >0.0 5)。　结论　外用rhEGF能明显促进深Ⅱ度烧伤创面愈合 ,加用肝素后效果更加显著 ,但早期使用rhEGF效果不明显     

Collagen in the scarless fetal skin wound: Detection with Picrosirius-polarization

Our group has developed an ovine model of deep dermal, partial-thickness burn where the fetus heals scarlessly and the lamb heals with scar. The comparison of collagen structure between these two different mechanisms of healing may elucidate the process of scarless wound healing. Picrosirius staining followed by polarized light microscopy was used to visualize collagen fibers, with digital capture and analysis. Collagen deposition increased with fetal age and the fibers became thicker, changing from green (type III collagen) to yellow/red (type I collagen). The ratio of type III collagen to type I was high in the fetus (166), whereas the lamb had a much lower ratio (0.2). After burn, the ratios of type III to type I collagen did not differ from those in control skin for either fetus or lamb. The fetal tissue maintained normal tissue architecture after burn while the lamb tissue showed irregular collagen organization. In conclusion, the type or amount of collagen does not alter significantly after injury. Tissue architecture differed between fetal and lamb tissue, suggesting that scar development is related to collagen cross-linking or arrangement. This study indicates that healing in the scarless fetal wound is representative of the normal fetal growth pattern, rather than a "response" to burn injury.     

重组人表皮生长因子促进大鼠皮肤创面愈合的研究

目的观察重组人表皮生长因子(rhEGF)对皮肤创面愈合的作用。方法制作大鼠背部创伤模型,采用自身平行对照,将34只大鼠背部的68个创面分成rhEGF治疗组与盐水对照组,观察大体形态和组织学改变、创面愈合时间和愈合率,测定伤后不同时间创面羟脯氨酸(OHP)含量和Ⅰ型Ⅲ型胶原比例,进行细胞DNA周期分析。结果经rhEGF治疗的创面愈合速度较盐水对照明显加快,2组平均愈合时间为(17.2±1.3)d和(20.5±1.6)d(P<0.01);外用rhEGF使创面肉芽组织生成增多,再上皮化明显,显著增加创面中OHP含量,降低Ⅰ型Ⅲ型胶原比例,加速细胞DNA复制。结论外用rhEGF可缩短创面愈合时间,增加肉芽组织及OHP含量,降低Ⅰ型Ⅲ型胶原比例,加速细胞DNA复制,明显促进皮肤创面的修复。     

Interaction of exogenous liposomal insulin-like growth factor-I cDNA gene transfer with growth factors on collagen expression in acute wounds

Growth factors have been shown to modulate the complex cascade of wound healing, however, interaction between different growth factors during dermal and epidermal regeneration is still not entirely defined. We have recently shown that exogenous liposomal gene transfer of cDNA results in physiologic expression and response in an acute wound. In the present study we determined the interaction between insulin-like growth factor-I (IGF-I), a mesenchymal growth factor, administered as liposomal cDNA, with other dermal and epidermal growth factors on collagen synthesis in an acute wound. Sprague-Dawley rats were given a scald burn to inflict an acute wound and divided into two groups to receive weekly subcutaneous injections of liposomes plus a beta-galactosidase containing plasmid (Lac Z [0.2 microg, vehicle]), or liposomes plus the IGF-I cDNA containing plasmid (2.2 microg) and Lac Z (0.2 microg). Immunological assays, histological and immunohistochemical techniques were used to determine growth factor concentration and different types of collagen (I, III, and IV) after IGF-I cDNA gene transfer. IGF-I cDNA transfer accelerated reepithelization and was associated with increased levels of IGF-I, fibroblast growth factor, keratinocyte growth factor, vascular endothelial cell growth factor, and platelet-derived growth factor protein expression. IGF-I cDNA had no effect on transforming growth factor-beta. IGF-I cDNA significantly increased type IV collagen while it had no effect on types I and III collagen. Exogenously administered IGF-I cDNA increased protein concentrations of keratinocyte growth factor, fibroblast growth factor, platelet-derived growth factor, and type IV collagen. We conclude that liposomal IGF-I gene transfer can accelerate wound healing without causing an increase in types I and III collagen expression.     

大鼠浅度烫伤创面愈合与相关生长因子及受体的基因表达

目的 探讨浅Ⅱ度烧伤创面愈合过程中相关生长因子及其受体的作用。方法 采用大鼠浅Ⅱ度烫伤模型,观察了大鼠浅Ⅱ度烫伤后创面愈合过程中组织学、炎性细胞、表皮细胞周期的变化,检测了创面相关生长因子及受体的基因表达。结果 炎性细胞到达创面依次为中性粒细胞、巨噬细胞和淋巴细胞;表皮细胞周期变化为伤后3天细胞增殖活跃,伤后7天细胞分裂明显增多;伤后1,3天PDGF（血小板源性生长因子）、PDGFR（血小板源性生长因子受体）和EGFR（表皮生长因子受体）基因表达明显增强,伤后3,5天EGF（表皮生长因子）、TGFβ－R2（转化生长因子β受体α）基因表达增强,伤后5,7天TGFβ－R1（转化生长因子β受体1）基因表达增强,伤后7,10天TGFβ1（转化生长因子β）基因表达增强。结论烧伤诱导相关生长因子及受体的基因表达,并且具有时相性和可逆性;相关生长因子基因表达与表皮细胞周期间可能存在相互调控作用,生长因子是调控创面愈合的主要因素。     

碱性成纤维细胞生长因子对大鼠烫伤创面中基质金属蛋白酶及其抑制剂的影响

目的　观察大鼠深Ⅱ度烫伤创面愈合过程中基质金属蛋白酶 (MMP) 2、MMP 7与金属蛋白酶组织抑制因子 2 (TIMP 2 )的变化以及创面外用碱性成纤维细胞生长因子 (bFGF)对其的影响。　方法　制作 30 %TBSA深Ⅱ度烫伤大鼠模型 ,分为单纯烫伤组和bFGF治疗组。于伤后 3、6h和 1、3、7、14d取创面皮肤标本 ,检测再上皮化率及胶原含量 ,并采用免疫组织化学染色法检测创面组织真皮内成纤维细胞MMP 2、MMP 7和TIMP 2的表达变化。另取 6只正常大鼠作为假烫组 ,分别检测上述各项指标。　结果　 (1)伤后 3～ 14d ,bFGF治疗组再上皮化率高于单纯烫伤组。 (2 )伤后 3h～ 3d ,bFGF治疗组和单纯烫伤组胶原含量持续下降 ,7～ 14d开始回升 ,但仍低于假烫组 (P<0 .0 5 )。 (3)伤后 1d ,单纯烫伤组MMP 2、MMP 7和TIMP 2表达增多 ,7d时达到高峰 ,持续到 14d。(4 )伤后 3～ 6h,bFGF治疗组MMP 2、MMP 7的表达与单纯烫伤组相似 ;伤后 1～ 14d ,3者的阳性表达强于单纯烫伤组。　结论　烫伤大鼠创面中细胞外基质的活动会影响皮肤的胶原沉积 ;MMP 2、MMP 7、TIMP 2的表达变化是组织修复的重要步骤 ,与bFGF加速创面愈合的过程密切相关。     

Liposomal gene transfer of keratinocyte growth factor improves wound healing by altering growth factor and collagen expression

BACKGROUND: Growth factors affect the complex cascade of wound healing; however, interaction between different growth factors during dermal and epidermal regeneration are still not entirely defined. In the present study, we thought to determine the interaction between keratinocyte growth factor (KGF) administered as liposomal cDNA with other dermal and epidermal growth factors and collagen synthesis in an acute wound. MATERIALS AND METHODS: Rats received an acute wound and were divided into two groups to receive weekly subcutaneous injections of liposomes plus the Lac-Z gene (0.22 microg, vehicle), or liposomes plus the KGF cDNA (2.2 microg) and Lac-Z gene (0.22 microg). Histological and immunohistochemical techniques were used to determine growth factor, collagen expression, and dermal and epidermal structure. RESULTS: KGF cDNA increased insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-3 (IGFBP-3), and fibroblast growth factor (FGF), decreased transforming growth factor-beta (TGF-beta), while it had no effect on platelet-derived growth factor (PDGF) levels in the wound. KGF cDNA significantly increased collagen Type IV at both the wound edge as well as the wound bed, while it had no effect on collagen Type I and III. KGF cDNA increased re-epithelialization, improved dermal regeneration, and increased neovascularization. CONCLUSIONS: Exogenous administered KGF cDNA causes increases in IGF-I, IGF-BP3, FGF, and collagen IV and decreases TGF-beta concentration. KGF gene transfer accelerates wound healing without causing an increase in collagen I or III.