TGFβ1在大黄多糖治疗糖尿病皮肤创面中的作用

目的探讨大黄多糖对糖尿病皮肤创面的疗效及可能机制。方法链脲佐菌素诱导糖尿病大鼠模型,2周后手术制作背部创面模型,再随机分为糖尿病组(给予生理盐水)、大黄多糖组(给予大黄多糖)及大黄多糖-SB431542组(给予大黄多糖及TGFβ1受体抑制剂SB431542混合液),另取正常大鼠制作背部创面模型给予生理盐水为对照组。2周后观测创面愈合率,检测创面羟脯氨酸含量,HE染色观测创面肉芽组织生长情况,免疫组化染色检测TGFβ1表达。结果与对照组相比,糖尿病组及大黄多糖-SB431542组创面愈合率明显降低,羟脯氨酸含量减少(均<0.01),而大黄多糖组无明显变化(>0.05)。对照组与大黄多糖组真皮层较厚,胶原纤维丰富,而糖尿病组与大黄多糖-SB431542组真皮层较薄,皮下胶原组织较少。糖尿病组TGFβ1表达较对照组减少,大黄多糖组及大黄多糖-SB431542组TGFβ1表达明显增加。结论诱导TGFβ1表达是大黄多糖促进糖尿病皮肤创面愈合的重要机制之一。     

组织工程全层皮肤修复糖尿病性皮肤溃疡创面的临床研究

目的探讨利用组织工程全层皮肤（ActivSkin）修复糖尿病性皮肤溃疡创面的临床应用方法。方法对皮肤溃疡患者采用控制血糖、增如营养、防治感染等措施，ActivSkin直接覆盖于创面，创面分3层包扎，常规换药，分别于术后观察组织工程皮肤的有效性及安全性。结果患者术后病情平稳，可见ActivSkin在创面存活，创面边缘可见白色新生组织形成，随时间延长创面逐渐缩小，平均28d后达到临床愈合。结论利用ActivSkin修复糖尿病性皮肤溃疡创面可促进创面早期修复、缩短病程和降低由此产生的致残率；为ActivSkin的临床应用提供了切实可行的方法。     

成纤维细胞生长因子在皮肤创面组织修复中的应用

皮肤是机体的重要屏障，烧伤后引起的局部或全身损害均与皮肤屏障的丧失有关，尽早修复缺损皮肤，封闭创面，重建皮肤屏障，有利于创面愈合和促进组织修复，最大限度地恢复皮肤的防御功能与外观。而目前大面积深度烧伤患者常存在严重的皮源不足，皮肤替代物的性能又较差，因此如何提高植皮成活率，改善皮肤替代物的性能就成为重要的因素。     

皮肤组织工程材料在深度烧伤创面修复中的应用

近年来迅速崛起的皮肤组织工程技术，正使深度烧伤创面的治疗发生着悄然的变化。它不仅改变着创面修复的传统理念，使我们逐步摆脱了对自体皮肤供区的依赖，简化了烧伤治疗过程，更影响着创面治疗的最终结局，使毁损皮肤结构和功能的真正修复成为了可能。纵观皮肤组织工程技术的发展历程，大致可分为三个阶段：表皮修复材料的构建、真皮修复材料的构建和全层皮肤修复材料的构建。作者拟将上述材料的特点及在深度烧伤创面修复中的应用情况作一简要评述并就未来发展方向作一简要展望。     

负载槲皮素的羧甲基壳聚糖水凝胶促进糖尿病大鼠创面愈合

背景：槲皮素具有抗炎、抗癌、抗氧化、抗衰老、抗抑郁等药理作用,具有极高的药用价值。槲皮素可以促进正常大鼠创面的愈合,但是以槲皮素为主要成分的药物研发较少,限制了其在临床的广泛应用。目的：以水凝胶材料负载槲皮素,探讨槲皮素-羧甲基壳聚糖水凝胶在糖尿病大鼠创面中的应用效果。方法：(1)分别制备羧甲基壳聚糖水凝胶与槲皮素-羧甲基壳聚糖水凝胶,表征水凝胶的微观形貌、体外释药性能。(2)细胞实验：将小鼠L929成纤维细胞分4组培养,空白对照组常规培养,纯水凝胶组、药物组、载药水凝胶组分别加入羧甲基壳聚糖水凝胶、槲皮素溶液与槲皮素-羧甲基壳聚糖水凝胶,检测细胞增殖与迁移能力。(3)动物实验：取80只SD大鼠建立糖尿病模型,造模成功后在大鼠背部制作直径2 cm的全层皮肤缺损创面,随机分4组干预,空白对照组、纯水凝胶组、药物组、载药水凝胶组创面分别注射生理盐水、羧甲基壳聚糖水凝胶、槲皮素溶液、槲皮素-羧甲基壳聚糖水凝胶,每组20只,无菌纱布包扎固定。术后观察创面愈合情况、创面病理形态、炎症因子表达及血管生成情况。结果与结论：(1)扫描电镜下可见两种水凝胶的微观形貌相似,均呈疏松多孔的网络结构,并且槲皮...     

硫化氢在ob/ob小鼠皮肤创面愈合中的作用与调控机制

目的:观察硫化氢(H2S)对ob/ob小鼠皮肤创面愈合的影响并探讨其作用机制。方法:将ob/ob小鼠随机分为生理盐水组、胰岛素组和NaHS(H2S供体)组,C57BL/6小鼠作为对照组,构建小鼠背部皮肤创面模型。干预后检测各组H2S释放量;用Western blot检测胱硫醚γ-裂解酶(CSE)及基质金属蛋白酶-9(MMP-9)蛋白的表达差异;用RT-qPCR检测CSE的mRNA表达变化;使用免疫组织化学法检测中性粒细胞及单核/巨噬细胞的浸润数量;使用ELISA检测肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6的水平;用Masson染色检测胶原沉积情况。结果:ob/ob小鼠皮肤创面肉芽组织中H2S释放及CSE蛋白、mRNA的表达水平以及胶原沉积显著低于C57BL/6小鼠(P0.05)。外源性H2S可加速ob/ob小鼠皮肤创面愈合(P0.05),增加胶原沉积。ob/ob小鼠创面中性粒细胞及单核/巨噬细胞浸润数量,TNF-α、IL-6的水平及MMP-9蛋白表达水平显著增加(P0.05),NaHS组显著降低。结论:H2S可显著改善糖尿病难愈性溃疡的愈合,作用机制可能与其抗炎作用有关。     

皮肤成纤维细胞在创面愈合中的研究进展

皮肤成纤维细胞是参与创面愈合的主要修复细胞,近年来其异质性及其与周围细胞间的通讯正逐渐引起重视.真皮成纤维细胞亚群主要包括乳头状成纤维细胞和网状成纤维细胞,对创面愈合发挥不同的作用.成纤维细胞通过自分泌和旁分泌信号分子与周围细胞之间相互作用构成创面微环境,影响创面愈合.慢性创面中的成纤维细胞表现出多种功能障碍.本文就成...     

环状RNA在皮肤相关疾病及创面愈合中的研究进展

环状RNA(circRNA)为一类具有连续共价闭环结构的新型非编码RNA(ncRNA),无3′及5′游离末端。其可作为微小RNA(miRNA)"分子海绵"调节转录或剪接,也可与RNA结合蛋白相互作用,在机体生理和病理过程中都起着关键作用。近来有研究表明,在与创面愈合相关的细胞(血管内皮细胞、角质细胞)增殖、迁移等活动中,有circRNA参与其中。本文回顾了circRNA在不同类型皮肤细胞中发挥的调节作用及其与皮肤相关疾病如皮肤鳞状细胞癌、基底细胞癌、重度痤疮的联系,旨在为阐释创面愈合的细胞分子机制提供新思路,以期为解决临床上创面治疗难题开拓新方向。     

外源性透明质酸对创面愈合影响的研究进展

对透明质酸参与创面修复过程及其机制的研究进展进行综述,回顾、总结与分析透明质酸盐的理化性质、代谢途径及代谢产物与创面修复的关系。透明质酸盐在创面修复过程中具有清创、抗炎、促进创伤愈合重要作用,其代谢产物能够促进血管生成及成纤维细胞增殖,调控胶原合成。透明质酸盐凭借在创面修复的重要作用,将会在创面修复及组织工程等领域得到广泛的应用。     

Heparan sulphate proteoglycan and wound healing in skin

Normal wound healing in skin embraces several reparative processes, many of which directly involve components of the extracellular matrix and the cutaneous basement membrane zone. Proteoglycans are a group of extracellular matrix macromolecules that have both structural and regulatory properties. In wound healing, certain proteoglycans fulfil a mechanical function of absorbing water and preventing tissue compression. However, proteoglycans may also have other roles in wound healing including a direct influence on inflammation, cell attachment and migration, and growth factor binding. Furthermore, proteoglycans may help to determine other aspects of the long-term quality of wound healing in skin through regulation of basement membrane permeability, epidermal hyperproliferation, and dermal fibrosis. © 1997 John Wiley & Sons, Ltd.     

miRNA在创伤愈合中的研究进展

微RNA(microRNA,miRNA)是近年来国内外生物医学领域的研究热点之一。作为一种含有21~25个核糖核苷酸的内源性非编码小分子单链RNA,miRNA通过与靶mRNA特异性的碱基互补配对,从而参与基因转录后的表达与调控,在多种疾病的发生发展过程中发挥重要功能。近来,越来越多的研究揭示miRNA参与了创伤愈合的全过程,深入阐释miRNA在不同阶段的病理生理干预机制,对于开展特定功能的靶向治疗具有重要意义。本文主要综述了部分miRNA在创伤愈合不同阶段中的调控研究进展。     

Hydrogen sulfide accelerates wound healing in diabetic rats

Aim: The aim of this study was to explore the role of hydrogen sulfide on wound healing in diabetic rats. Methods: Experimental diabetes in rats was induced by intraperitoneal injection of streptozotocin (STZ) (in 0.1 mol/L citrate buffer, Ph 4.5) at dose of 70 mg/kg. Diabetic and age-matched non-diabetic rats were randomly assigned to three groups: untreated diabetic controls (UDC), treated diabetic administrations (TDA), and non-diabetic controls (NDC). Wound Healing Model was prepared by making a round incision (2.0 cm in diameter) in full thickness. Rats from TDA receive 2% sodium bisulfide ointment on wound, and animals from UDC and NDC receive control cream. After treatment of 21 days with sodium bisulfide, blood samples were collected for determination of vascular endothelial growth factor (VEGF), intercellular cell adhesion molecule-1 (ICAM-1), antioxidant effects. Granulation tissues from the wound were processed for histological examination and analysis of western blot. Results: The study indicated a significant increase in levels of VEGF and ICAM-1 and a decline in activity of coagulation in diabetic rats treated with sodium bisulfide. Sodium bisulfide treatment raised the activity of superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) protein expression, and decreased tumor necrosis factor α (TNF-α) protein expression in diabetic rats. Conclusions: The findings in present study suggested that hydrogen sulfide accelerates the wound healing in rats with diabetes. The beneficial effect of H2S may be associated with formation of granulation, anti-inflammation, antioxidant, and the increased level of vascular endothelial growth factor (VEGF).     

Combined effects of artificial dermis and vascular endothelial growth factor concentration gradient on wound healing in diabetic porcine model

Wounds in patients with diabetes mellitus are one of the most prevalent impaired wounds in the world. Vascular endothelial growth factor (VEGF) is one of the most important proangiogenic mediators. Artificial dermal (AD) such as Pelnac^® has been shown, in humans and animal models, a great therapeutic potential in full-thickness skin wounds. We attempt to promote the wound healing in diabetic porcine models through combined use of AD and constant concentration of VEGF or VEGF concentration gradient. We created full-thickness excisional wounds in diabetic animal models. Analyzed the healing process through images, histology and immunohistochemistry. Results show that the combination of AD and concentration gradient of VEGF could provide an appropriate angiogenesis, improve granulation formation, increase epithelization and maintain the VEGF levels of the wound bed. Eventually accelerate the direct healing of diabetic wounds or make good preparation for secondary skin graft.     

Antibodies to wounded tissue enhance cutaneous wound healing

The wound repair process is a highly ordered sequence of events that encompasses haemostasis, inflammatory cell infiltration, tissue regrowth and remodelling. Wound healing follows tissue destruction so we hypothesized that antibodies might bind to wounded tissues, which would facilitate the engulfment of damaged tissues by macrophages. Here, we show that B cells, which produce antibodies to damaged tissues, are engaged in the process of wound healing. Splenectomy delayed wound healing, and transfer of spleen cells into splenectomized mice recovered the delay in wound healing. Furthermore, wound healing in splenectomized nude mice was also delayed. Transfer of enriched B220⁺ cells by magnetic beads accelerated wound healing in splenectomized mice. We detected immunoglobulin G1 (IgG1) binding to wounded tissues by using fluorescein isothiocyanate-labelled anti-IgG1 6–24 hr after wounding. Splenectomy reduced the amount of IgG1 binding to wounded tissues. Immunoblotting studies revealed several bands, which were reduced by splenectomy. Using immunoprecipitation with anti-IgG bound to protein G we found that the intensity of several bands was lower in the serum from splenectomized mice than in that from sham-operated mice. These bands were matched to myosin IIA, carbamoyl-phosphate synthase, argininosuccinate synthase, actin and α-actinin-4 by liquid chromatography tandem mass spectrometry analysis.     

Identification of a reference gene for the quantification of mRNA and miRNA expression during skin wound healing

Aim: Wound healing is a coordinated process to restore tissue homeostasis and reestablish the protective barrier of the skin. miRNAs may modulate the expression of target genes to contribute to repair processes, but due to the complexity of the tissue it is challenging to quantify gene expression during the distinct phases of wound repair. Here, we aimed to identify a common reference gene to quantify changes in miRNA and mRNA expression during skin wound healing. Methods: Quantitative real-time PCR and bioinformatic analysis tools were used to identify suitable reference genes during skin repair and their reliability was tested by studying the expression of mRNAs and miRNAs. Results: Morphological assessment of wounds showed that the injury model recapitulates the distinct phases of skin repair. Non-degraded RNA could be isolated from skin and wounds and used to study the expression of non-coding small nuclear RNAs during wound healing. Among those, RNU6B was most constantly expressed during skin repair. Using this reference gene we could confirm the transient upregulation of IL-1β and PTPRC/CD45 during the early phase as well as the increased expression of collagen type I at later stages of repair and validate the differential expression of miR-204, miR-205, and miR-31 in skin wounds. In contrast to Gapdh the normalization to multiple reference genes gave a similar outcome. Conclusion: RNU6B is an accurate alternative normalizer to quantify mRNA and miRNA expression during the distinct phases of skin wound healing when analysis of multiple reference genes is not feasible.     

Sonic hedgehog促1型糖尿病小鼠的伤口愈合

目的：研究sonic hedgehog（Shh）及其受体Ptc1在糖尿病小鼠伤口愈合中的作用。方法：分别在正常和链脲佐菌素（STZ）诱导的糖尿病小鼠建立皮肤损伤模型,Western印迹检测Shh和Ptc1的蛋白水平;观察外源性Shh或Ptc1抑制剂cyclopamine对伤口愈合的影响。结果：（1）正常小鼠损伤后皮肤组织中的Shh和Ptcl蛋白质表达明显升高;外源性Shh对伤口愈合无明显促进作用,但cyclopamine可以明显地抑制伤口愈合;（2）STZ诱导的糖尿病小鼠,其皮肤组织中内源性的Shh和Ptcl蛋白水平明显下调;（3）外源性Shh可显著促进糖尿病小鼠伤口的愈合,且呈浓度依赖性;Cyclopamine则明显地抑制糖尿病小鼠的伤口愈合。结论：Shh-Ptc1通路参与了皮肤伤口愈合,糖尿病伤口愈合延迟与Shh-Ptc1表达下调有关。     

Basal membrane heparan sulphate proteoglycan expression during wound healing in human skin

Heparan sulphate proteoglycans (HSPGs) are integral components of the basement membrane (BM) in various tissues. HSPGs are important in the assembly and structure of the BM, and their putative functions include regulation of basement membrane permeability, binding of growth factors, and a role in cellular adhesion. In this study the expression of HSPG was examined during wound healing in human skin, using monoclonal antibodies (MAbs) that recognize the HSPG core protein and two different heparan sulphate (HS) epitopes, and the dynamics of HSPG expression were investigated in relation to epidermal cellular proliferation and permeability of the BM. Healing of excisional wounds in healthy volunteers was studied from day 0 up to 1 year. Intact human skin showed strong continuous staining of the dermo-epidermal BM and the vascular BM with all MAbs. Up to day 4 after wounding, staining for HSPG was absent under the ingrowing epidermis, with any of the MAbs, indicating that no complete BM was present. From day 7 onwards, the BM of the neo-epidermis showed positive staining for the HSPG core protein and a low sulphated HS epitope, and after day 14, the staining intensity was similar to normal skin. The staining patterns of these HSPG epitopes were similar to that of laminin. The staining pattern with a MAb against an epitope in the highly sulphated part of HS was found to be distinct from the other BM markers studied. This epitope was absent under the neo-epidermis up to 2 months after wounding. One year after wounding, the epitope was found to be present again. We observed that only in the time period between 2 months and 1 year had the epidermis normalized with respect to the number of cycling cells and the absence of high molecular weight plasma proteins. These findings suggest a correlation between normalization of epidermal proliferation, BM permeability, and regeneration of BM HS. It is proposed that complete BM maturation following skin wounding is a slow process and may account for the epidermal abnormalities that persist for a considerable period of time after wound healing. © 1997 John Wiley & Sons, Ltd.     

组织工程化人工皮肤的构建与应用

体外培养表皮细胞构建组织工程化人工皮肤是促进皮肤缺损创面愈合、提高创面修复质量的新途径。本文就人工皮肤种子细胞研究现状、真皮支架材料的研制、复合人工皮肤的构建与应用作一综述     

Fibronectin and tenascin in rat tracheal wound healing and their relation to cell proliferation

To investigate the relationship between cell proliferation and distribution of fibronectin and tenascin during wound healing, light and electron microscopy and immunohistochemistry for fibronectin, tenascin, and 5-bromodeoxyuridine (BrdU) were performed following mechanical injury of rat trachea. Tenascin staining appeared 18h after curettage, when the percentage of BrdU-positive nuclei was maximal in the epithelium. Once tenascin appeared, the labeling index of BrdU-positive epithelial nuclei decreased rapidly. Distribution of tenascin was restricted to granulation tissue In curetted areas which were covered with regenerating epithelium, while fibronectin stained diffusely in both curetted and non-curetted areas. Analysis of the relative intensity of fibronectin and tenascin staining showed that decreases of fibronectin staining were followed by increasing tenascin staining. It is proposed that fibronectin and tenascin may contribute differently to tissue repair in the trachea by interfering with cell proliferation of epithelial cells and fibroblasts.