电刺激糖尿病模型大鼠创面的愈合及血管生成

文题释义：糖尿病性难愈创面：是最常见的糖尿病并发症之一,主要由于体内高糖环境和多种生物学因素共同作用导致创面形成,大多难以治愈,以糖尿病足为代表,多数患者面临着截肢风险,其主要机制有晚期糖基化终末产物相关学说、炎性浸润、外周神经功能障碍等。 血管生成：指从已有的毛细血管或毛细血管后静脉发展而形成新的血管,主要包括：激活期血管基底膜降解;血管内皮细胞的激活、增殖、迁移;重建形成新的血管和血管网,是一个涉及多种细胞的多种分子的复杂过程。 背景：糖尿病性创面愈合障碍是糖尿病的常见并发症之一,然其发病机制目前尚未阐明。电刺激疗法是临床治疗创面损伤的常用手段之一,能有效促进损伤皮肤愈合。 目的：探讨电刺激疗法对糖尿病大鼠创面愈合及血管生成的影响及潜在机制。 方法：SPF级雄性SD大鼠42只,随机分为空白对照组和糖尿病模型组。模型组采用一次性尾静脉注射链脲佐菌素(50 mg/kg)联合背部皮肤创伤的方法建立糖尿病性难愈创面大鼠模型,再将造模成功的24只大鼠随机分为模型组和电刺激治疗组;空白对照组为正常大鼠造成背部创面,治疗组予以电刺激治疗21 d,对照组和模型组正常饲养,于治疗第3,7,14,21天分别评价创面愈合情况。治疗完成后,取大鼠血清和创面组织用于指标检测,采用苏木精-伊红染色法观察大鼠创面病理形态,酶联免疫吸附法检测血清内皮型一氧化氮合酶、血管生成素1和血管内皮生长因子水平,免疫组织化学法和Western-blotting法定位定量观察创面血管生成素受体2和血管内皮细胞生长因子受体2蛋白的表达。实验方案经西南医科大学附属医院动物实验伦理委员会批准。结果与结论：①经电刺激治疗后,糖尿病大鼠创面愈合率于第14天时即接近90%,而模型组不到60%,差异有显著性意义(P < 0.01);血清内皮型一氧化氮合酶、血管生成素1和血管内皮生长因子水平较模型组均显著升高(P < 0.05或P < 0.01);②与模型组比较,治疗组创面新生血管多,血管管腔较大,血管周围及创面组织的血管生成素受体2和血管内皮细胞生长因子受体2表达均显著升高(P < 0.05或P < 0.01);③电刺激疗法能明显促进糖尿病大鼠创面愈合及新血管生成,其机制与升高内皮型一氧化氮合酶、血管生成素1和血管内皮生长因子水平,上调血管生成素受体2和血管内皮细胞生长因子受体2水平有关。 ORCID: 0000-0001-9624-6694(耿康) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

皮肤发育相关的miRNAs及其在糖尿病创面愈合中的作用*

微小核糖核酸（microRNA/miRNA）作为一类进化上保守的非编码小分子RNA,参与基因转录后的表达与 调控,其表达模式有一定的时间性和空间性,体现在不同的miRNAs在不同组织、不同发育阶段的表达水平差异。 某些miRNAs能够促进创面愈合,在创面愈合的炎症期抑制炎性介质的表达;某些miRNAs能够促进增生期创面 细胞的增殖、迁移,有利于创面的快速修复;在创面重塑期,某些miRNAs又能够通过抑制无痕愈合信号通路的 相关蛋白质来促使瘢痕修复。糖尿病创面通常伴随糖尿病周围神经病变、糖尿病血管病变和感染。部分miRNAs 通过调控特定基因的表达水平,激活或抑制不同且特定的信号通路,一定程度上促进了糖尿病创面的愈合。本文 主要综述了miRNA在创面愈合过程不同阶段的调控研究进展,以及miRNAs促进、抑制糖尿病创面愈合的机制, 以期为后续研究开拓新的思路。     

自噬对烧伤创面愈合的影响

细胞自噬是细胞在应激状态下应用溶酶体对自身损伤细胞器等物质进行分解,将产生的大分子物质予以回收利用,从而保留细胞活性的降解过程。自噬在细胞的发育和分化进程中起着至关重要的作用。烧伤创面经历炎症期、细胞增殖期和组织重塑期3个时期完成组织愈合。细胞自噬在烧伤创面愈合过程中,具有促进病原体清除,参与新生血管增殖、肉芽组织形成,改善上皮的角质化以及瘢痕重塑的作用。调控自噬,可影响烧伤创面愈合。本文综述细胞自噬在烧伤创面愈合中的研究进展。     

活化素受体样激酶1及其在血管生成中的作用

正血管生成是指新的血管从已存在的毛细血管网生成的过程,受血管生成促进因子和抑制因子的严格调控。通常,血管生成发生于胚胎和出生后早期血管的发育过程中,除女性生理周期和伤口愈合等过程外,在成年阶段血管生成已处于静息状态。但是,在发生某些疾病如肿瘤、糖尿病视网膜病变、心血管疾病及类风湿性关节炎[1-2]时,血管生成过程被异常激活。活化素受体样激酶1(activin receptor-like     

血管生成相关因子与瘢痕增生

背景:瘢痕是创伤修复的产物,严重的瘢痕增生常常导致机体的畸形及功能障碍。血管生成伴随着瘢痕形成的整个过程,与瘢痕的形成密切相关。细胞因子也参与了与瘢痕的形成的过程。目的:关于血管生成相关细胞因子与瘢痕增生进行阐述,了解血管生成相关细胞因子通过影响瘢痕形成过程中血管的生成与成熟,影响瘢痕的增生及萎缩成熟,最终为临床防治瘢痕提供新的理论依据及方法。方法:应用计算机检索中文数据库CNKI,应用"瘢痕,血管生成,细胞因子"等为检索词进行检索;应用计算机检索英文数据库Pub Med及Medline,应用"scar,angiogenesis,cytokine"等为检索词进行检索。筛选关于瘢痕增生与血管生成,瘢痕增生与细胞因子,血管生成与细胞因子的相关文献,最后筛选纳入54条文献。结果与结论:瘢痕的形成是机体各种复杂因素综合作用的结果,受多种因素的影响。瘢痕的形成过程中存在血管生成和成熟。各种血管生成相关因子必然会通过影响血管生成成熟,从而影响瘢痕的增生和萎缩。在临床应用中通过干预相关血管生成因子的表达或阻断其作用,影响瘢痕组织中的血管生成与成熟,从而达到防治瘢痕的目的,为瘢痕治疗与预防提供新的思路及方法。     

生长激素/生长因子对微血管生成的影响的研究进展

血管生成(angiogenesis)是从先存血管产生新血管的过程,血管生成可发生在伤口愈合、子宫内膜周期性变化、肿瘤、心肌梗塞后、糖尿病眼底改变和肾损害[1]。正是确认了外源性生长激素可导致视网膜血管增生,人们才发现生长激素可用于某些缺血性疾病。现就生长激素与生长因子在血管生成初始的关系,以及在血管内皮细胞去分化和各种条件下最后形成新的血管的机制进行综述。1　生长激素引起微血管生成的机制在使用生长激素治疗的非糖尿病患者(因下丘脑外伤需外源性生长激素)身上发现视网膜血管新生[2],发现这一现象之初,研究者假设是高血糖所致,但…     

皮肤感觉神经与创面组织相互作用的研究

【摘要】皮肤感觉神经对创面愈合过程中的炎性介质反应、新血管形成、肉芽增生和愈合后重塑均有重要调控作用,同时创面的组织细胞又能反作用于皮肤感觉神经。对皮肤感觉神经的合理干预能够有效改善创面的转归。     

单核/巨噬细胞在血管生成和侧枝血管生长中的作用

血 管生成 (Ａｎｇｉｏｇｅｎｅｓｉｓ)是指毛细血管的增生和发芽 ,即血管形成 ,包括基质溶解、细胞转移、粘附、增生和管腔形成 ;侧枝血管生长 (ＣｏｌｌａｔｅｒａｌＶｅｓｓｅｌＧｒｏｗｔｈ)是指已有的连接侧枝动脉的微动脉原位增生形成肌型动脉 ,或者原有的侧枝血管形成。虽然毛细血管的形成可缓解缺血组织的灌注不足 ,但只有侧枝动脉能提供足够的血液到缺血区[1] 。在缺血区二种血管生长方式都存在 ,在不同的部位和条件常以一种生长方式为主[1,2 ] 。单核细胞系统的细胞在成人主要来源于骨髓造血干细胞 ,骨髓中的髓样干细胞受细胞因子…     

创面愈合及瘢痕形成中的结缔组织生长因子

背景:创面愈合是外科学研究的热点。结缔组织生长因子参与调节皮肤创面愈合过程并发挥了重要作用。目的:综述结缔组织生长因子在创面中的表达规律及其在瘢痕愈合和创面愈合中的作用。方法:运用计算机检索中国CNKI学术总库(www.cnki.net)、万方数据库、维普中文科技期刊数据库、Springer Link及Pub Med数据库相关文章。以"结缔组织生长因子;瘢痕;创面愈合;再生"为中文检索词;以"CTGF;CCN2;Wound healing;Regeneration"为英文检索词。纳入37篇关于结缔组织生长因子的基础研究及其在创面愈合实验中相关研究进行讨论。结果与结论:结缔组织生长因子在皮肤创面愈合及瘢痕形成过程中呈动态表达。在创伤修复初期,结缔组织生长因子的短暂上调可促进肉芽组织和新生血管形成等加速创面修复过程;在伤口修复和重塑阶段的持续表达可明显促进过度基质的产生、胶原沉积和瘢痕挛缩,导致病理性瘢痕的发生。结缔组织生长因子参与了皮肤创面的愈合过程,其在加速创面愈合和控制瘢痕形成方面可能具有应用前景。结缔组织生长因子作为转化生长因子β的下游产物,除本身的生物学作用外,其介导转化生长因子β的促成纤维细胞增殖、促纤维化、诱导黏附及迁移等作用是其影响创面愈合的作用机制之一。     

深Ⅱ度烧伤愈合后Bcl-xl、Bax蛋白表达与增生性瘢痕的相关性

目的探讨Bcl-xl和Bax蛋白在深Ⅱ度烧伤愈合后不同时期增生性瘢痕中的表达特点。方法人深Ⅱ度烧伤愈合后不同时期的增生性瘢痕皮肤40例,正常皮肤组织10例。分为增生期组、减退早期组、减退晚期组、成熟期组,正常对照组。免疫组织化学染色,检测Bcl-xl和Bax蛋白的表达。结果深Ⅱ度烧伤创面愈合后,Bcl-xl和Bax蛋白主要表达于表皮基底细胞和真皮层成纤维细胞,增生期和减退早期Bcl-xl和Bax蛋白产物光密度值明显高于正常皮肤(p<0.01),减退晚期和成熟期随瘢痕成熟而逐渐递减接近正常皮肤。结论 Bcl-xl和Bax蛋白表达与深Ⅱ度烧伤创面愈合后增生性瘢痕的发生和瘢痕成熟相关。     

血管新生的分子机制与相关疾病

正血管新生指从已存在的血管上生长出新的毛细血管的过程~([1])。该过程十分复杂,依赖血管内皮细胞、成纤维细胞、巨噬细胞和细胞外基质的相互作用,并受促血管新生分子和抗血管新生分子协同调控。生理条件下,促血管新生分子和抗血管新生分子保持动态平衡;一旦平衡被打破,机体就会出现多种疾病,如肿瘤、多种眼科疾病、帕金森病(Parkinson’s disease,PD)和阿尔茨海默症(Alzheimer’s di-     

早期磨痂后生物皮覆盖修复深Ⅱ度烧伤创面136例报告

目的观察早期磨痂术后生物皮覆盖修复深Ⅱ度烧伤创面的效果。方法136例创面分别于伤后1～2d内，在全麻无菌技术下用消毒钢丝球或电动磨削机彻底磨去创面坏死痂皮组织，然后用生物皮覆盖，观察创面愈合时间及瘢痕增生情况。创面愈合后1～1．5年随访观察。结果130例创面Ⅰ期愈合，Ⅰ期愈合率96．0％，6例因生物皮出现部分液化感染，创面延期愈合。愈合时间为13～20d，平均（15±2）d。随访130例肤色基本正常，4例出现轻度瘢痕增生，2例出现花斑样色素沉着。结论早期磨痂能有效清除仓4面坏死组织，降低烧伤毒素的产生，全身炎症反应较轻；生物皮完全无菌，与创面粘附性不变，抗原性减小，可以有效保护创面，使创面愈合快，瘢痕增生少，是一种十分理想的创面覆盖物。     

Cytotoxicity and immunogenicity of SACCHACHITIN and its mechanism of action on skin wound healing

SACCHACHITIN membrane, a weavable skin substitute made from the residual fruiting body of Ganoderma tsugae, has been demonstrated to promote skin wound healing. Prior to its clinical application, it is critical to learn more about any possible cytotoxicity, immunogenicity, or allergy response, and at least some of its mechanism(s) of action(s). In the present studies, it has been found that SACCHACHITIN suspension at less than 0.05% shows no cytotoxicity to the primary culture of rat fibroblasts. However, at higher concentrations (> or = 0.1%), it does reduce the growth of fibroblasts, based on MTT assays. This might be caused by positive charges on chitin molecules that are too strong, and may be harmful to the cell membrane. SACCHACHITIN showed no immunogenicity after it was inoculated into rats three times; however, the unmodified, purified rabbit type I and type II collagens did. Subcutaneous injection of SACCHACHITIN suspension into rats showed no gross allergic responses on skin. Nevertheless, it did cause local acute inflammation, as observed by histological investigation. This is similar to what occurred in the wound site covered with SACCHACHITIN membrane. The chemotactic effect of SACCHACHITIN was exhibited in both intact and wounded skin tissues. This may be one of the initial beneficial effects of SACCHACHITIN membrane to wound healing. The rapid acute inflammatory process was followed by the appearance of angiogenesis and granulation tissue formation, which occurred earlier than it normally would. Coverage of the wound area with SACCHACHITIN membrane also induced an earlier formation of scar tissue to replace the granulation tissue. A 1.5 x 1.5 cm(2) wound area covered by SACCHACHITIN completely healed by 21 days, while that covered with cotton gauze did not. Therefore, SACCHACHITIN is a safe biomaterial for use as a wound dressing for skin healing. Its promoting action for wound healing might be due to its chemotactic effect for inflammatory cells. This, in turn, may facilitate subsequent angiogenesis, granulation tissue formation, and faster new tissue formation, leading to faster wound healing.     

瘢痕组织中α-平滑肌肌动蛋白的表达与细胞凋亡的关系

目的:观察α-平滑肌肌动蛋白在瘢痕组织中的表达,了解病理性瘢痕形成过程中凋亡所扮演的角色及其与肌成纤维细胞在真皮中变化的关系。方法:瘢痕标本来自烧伤后来我院进行整形手术的病人,同时取病人手术供皮区的正常皮肤作为对照。 8例瘢痕组织标本 (含 2例愈合较为平坦的瘢痕和 6例增生性瘢痕组织)被分成增殖期和成熟期两组。运用caspase-3mRNA及其蛋白的表达及TUNEL方法检测瘢痕及正常组织中的凋亡细胞,并以免疫组化法检测瘢痕及正常皮肤真皮内α-平滑肌肌动蛋白单克隆抗体的表达。结果:瘢痕组织中细胞凋亡的数目与正常组织明显不同。瘢痕内的TUNEL标记阳性细胞数多于正常组织;增殖期瘢痕内的细胞凋亡的数目多于成熟期。增殖期TUNEL标记阳性的细胞多于平坦瘢痕,而成熟期两者无显著差别,Caspase-3mRNA及其蛋白的表达与TUNEL标记结果具有一致性。随着瘢痕组织的成熟,α-平滑肌肌动蛋白单克隆抗体的表达逐渐降低,平坦的瘢痕组织中的表现尤为明显;增生性瘢痕中,增殖期与成熟期之间无显著差别。结论:正常伤口愈合过程中,肌成纤维细胞暂时性的表达,可引起伤口的收缩,随着真皮再塑形,含有α-平滑肌肌动蛋白的肌成纤维细胞因凋亡而消失,而病理性的愈合结局可能是它持续表达的结果。     

Scar‐free cutaneous wound healing in the leopard gecko,Eublepharis macularius

Cutaneous wounds heal with two possible outcomes: scarification or near‐perfect integumentary restoration. Whereas scar formation has been intensively investigated, less is known about the tissue‐level events characterising wounds that spontaneously heal scar‐free, particularly in non‐foetal amniotes. Here, a spatiotemporal investigation of scar‐free cutaneous wound healing following full‐thickness excisional biopsies to the tail and body of leopard geckos (Eublepharis macularius) is provided. All injuries healed without scarring. Cutaneous repair involves the development of a cell‐rich aggregate within the wound bed, similar to scarring wounds. Unlike scar formation, scar‐free healing involves a more rapid closure of the wound epithelium, and a delay in blood vessel development and collagen deposition within the wound bed. It was found that, while granulation tissue of scarring wounds is hypervascular, scar‐free wound healing conspicuously does not involve a period of exuberant blood vessel formation. In addition, during scar‐free wound healing the newly formed blood vessels are typically perivascular cell‐supported. Immunohistochemistry revealed widespread expression of both the pro‐angiogenic factor vascular endothelial growth factor A and the anti‐angiogenic factor thrombospondin‐1 within the healing wound. It was found that scar‐free wound healing is an intrinsic property of leopard gecko integument, and involves a modulation of the cutaneous scar repair program. This proportional revascularisation is an important factor in scar‐free wound healing.     

Tissue Engineering and Regenerative Repair in Wound Healing

Wound healing is a highly evolved defense mechanism against infection and further injury. It is a complex process involving multiple cell types and biological pathways. Mammalian adult cutaneous wound healing is mediated by a fibroproliferative response leading to scar formation. In contrast, early to mid-gestational fetal cutaneous wound healing is more akin to regeneration and occurs without scar formation. This early observation has led to extensive research seeking to unlock the mechanism underlying fetal scarless regenerative repair. Building upon recent advances in biomaterials and stem cell applications, tissue engineering approaches are working towards a recapitulation of this phenomenon. In this review, we describe the elements that distinguish fetal scarless and adult scarring wound healing, and discuss current trends in tissue engineering aimed at achieving scarless tissue regeneration.     

Adenosine promotes wound healing and mediates angiogenesis in response to tissue injury via occupancy of A(2A) receptors

Recent evidence indicates that topical application of adenosine A(2A) receptor agonists, unlike growth factors, increases the rate at which wounds close in normal animals and promotes wound healing in diabetic animals as well as growth factors, yet neither the specific adenosine receptor involved nor the mechanism(s) by which adenosine receptor occupancy promotes wound healing have been fully established. To determine which adenosine receptor is involved and whether adenosine receptor-mediated stimulation of angiogenesis plays a role in promotion of wound closure we compared the effect of topical application of the adenosine receptor agonist CGS-21680 (2-p-[2-carboxyethyl]phenethyl-amino-5'-N-ethylcarboxamido-adenosine) on wound closure and angiogenesis in adenosine A(2A) receptor knockout mice and their wild-type littermates. There was no change in the rate of wound closure in the A(2A) receptor knockout mice compared to their wild-type littermates although granulation tissue formation was nonhomogeneous and there seemed to be greater inflammation at the base of the wound. Topical application of CGS-21680 increased the rate of wound closure and increased the number of microvessels in the wounds of wild-type mice but did not affect the rate of wound closure in A(2A) receptor knockout mice. Similarly, in a model of internal trauma and repair (murine air pouch model), endogenously produced adenosine released into areas of internal tissue injury stimulates angiogenesis because there was a marked reduction in blood vessels in the walls of healing air pouches of A(2A) receptor knockout mice compared to their wild-type controls. Inflammatory vascular leakage and leukocyte accumulation in the inflamed air pouch were similarly reduced in the A(2A) receptor knockout mice reflecting the reduced vascularity. Thus, targeting the adenosine A(2A) receptor is a novel approach to promoting wound healing and angiogenesis in normal individuals and those suffering from chronic wounds.     

COX-2信号通路及其调控对皮肤伤口愈合的干预研究

目的 通过体外及体内干预实验研究探讨COX-2信号通路对皮肤伤口炎症细胞、新生血管的影响,评价皮肤伤口愈合的情况,为减少伤口瘢痕形成、促进愈合的诊疗方法提供实验依据.方法 采用体外细胞培养划痕试验及小鼠体内实验,运用COX-2选择性抑制剂尼美舒利进行干预,通过伤口皮肤铺片、免疫组化对伤口生长过程中炎症细胞、新生血管、成...     

选择性环氧合酶2抑制剂尼美舒利抑制皮肤瘢痕形成的研究

背景：尼美舒利是一种选择性环氧合酶2抑制剂,可通过抑制环氧合酶2的合成、抑制前列腺素的合成、白细胞的介质释放等环节影响炎症的发生过程,具有解热、镇痛和抗炎作用。 目的：探讨尼美舒利抑制小鼠皮肤瘢痕形成的作用及机制。 方法：昆明种小鼠背部脊柱两侧建立皮肤损伤模型,随机分为模型组、25,50,100 μmol/L尼美舒利组,切口分别涂抹0.5 mL玻璃酸钠凝胶、25,50,100 μmol/L尼美舒利玻璃酸钠分散制剂。 结果与结论：应用25,50 μmol/L尼美舒利可以明显减小瘢痕组织的面积(P < 0.01),但100 μmol/L尼美舒利却造成损伤面积增大(P < 0.05),与模型组相比,不同剂量的尼美舒利治疗的小鼠在创伤形成第3,7,14天环氧合酶2的表达明显降低,且其中50和100 μmol/L尼美舒利治疗的小鼠创口微血管的生成速度明显降低(P < 0.05),提示尼美舒利可以通过适度下调皮肤修复过程中环氧合酶2的表达,控制微血管的形成,达到抑制病理性瘢痕形成的治疗目的,但剂量过大(100 μmol/L)则可抑制创伤的愈合。