离体冻伤模型创面愈合过程中基质细胞衍生因子1促表皮干细胞迁移的研究

目的 了解离体皮肤冻伤模型创面愈合过程中,基质细胞衍生因子1(SDF-1)对创缘表皮干细胞(ESC)的运动趋化作用.方法 体外构建三维皮肤等价物全层冻伤模型(创面呈孔形),免疫组织化学染色观察冻伤后3、7 d创缘间质内SDF-1的表达情况.另将冻伤模型分为对照组(创面区添加PBS 50 μL/孔)、SDF-1组(创面区添加100 ng/mL SDF-1,50μL/孔)和AMD3100组[创面区用100 ng/mL AMD3100(50μL/孔)处理30 min后,添加SDF-1 50μL/孔],观察各组创缘中ESC的分布变化.结果 免疫组织化学染色显示,冻伤后3、7 d创缘间质内SDF-1的表达逐渐增加.与对照组及AMD3100组相比,SDF-1组创缘基底层整合素β1阳性细胞数量增多,且部分阳性细胞向上层表皮迁移聚集,创缘ESC数量更多,表皮细胞层数增加.结论 SDF-1促进ESC向创缘迁移聚集参与创面修复,这可能是ESC参与创面修复过程的机制之一.     

创面愈合过程中创缘表皮干细胞的异位

目的观察创缘表皮干细胞在全层皮肤创面愈合过程中的分布特征,初步探讨其在创面愈合过程中的作用.方法将已行BrdU活体标记的 20只Wistar大鼠背部制备4个为2.54 cm2的全层皮肤创面,分别于伤后3、7、14和21 d(n=5)行组织学检查,动态观察创面愈合情况,并行β1整合素、角蛋白19(K19)与BrdU免疫组织化学法检测表皮干细胞在创面愈合过程中的分布情况.结果创面愈合率为83.75%(61/80).所有创面肉芽组织于各时相点均未见β1整合素、K19阳性细胞出现,但于创缘表皮的棘层或颗粒层均出现散在的β1整合素、K19和BrdU阳性细胞.且越接近创面阳性细胞越密集,组织学上与基底层的阳性细胞无直接联系;其数量随创面的缩小逐渐增加,直至创面愈合.创面上皮化后,阳性细胞逐渐减少,并随愈合创面表皮脚的出现而消失.而感染创面的阳性细胞数量明显少于未感染创面.结论表皮干细胞能主动参与创面的修复,创缘的表皮干细胞异位的主要功能可能是促进创面再上皮化.     

创面愈合过程中创缘表皮干细胞的再分布

目的研究皮肤干细胞在全层皮肤创面愈合过程中的分布与增殖分化特征,以及该特征在创面愈合过程中的作用.方法10只Wistar大鼠背部各制备4个面积为2.54cm2的全层皮肤创面,将刨面随机分为2组,即银锌霜治疗组(20个创面),空白对照组(20个创面).分别于伤后3d、1周、2周和3周以组织学检查动态观察各组治疗效果,并以β1整合素、角蛋白19(K19)免疫组化法检测表皮干细胞在创面愈合过程中的表达情况.结果两组创面愈合率为银锌霜组80%(16/20),对照组60%(12/20).两组创面肉芽组织于各时相点均未见β1整合素、K19阳性细胞出现,但于创缘表皮的棘层或颗粒层出现了散在的β1整合素和K19同时染色阳性细胞.且越接近创面这些阳性细胞越密集,组织学上与基底层的阳性细胞无直接联系,其数量随着创面的缩小渐渐增加,直到创面愈合.上皮化后,这些阳性细胞逐渐减少,并随着愈合创面表皮脚的出现而消失,而感染创面的阳性细胞数量明显少于未感染创面的阳性细胞数.结论表皮干细胞能动地参与创面的修复,创缘表皮干细胞再分布的主要功能可能促进创面再上皮化.     

Np63α在创面愈合过程中表达改变的研究

目的：探讨p63基因编码的蛋白亚型△Np63α在创面愈合表皮再生中的表达及其生物学意义。方法：①在3周龄小鼠背部用直径1．6cm的环钻压切制备出一个2．0cm~2全层皮肤缺损的圆形创面,并将切除的皮肤原位缝合以覆盖创面。②将18只昆明小鼠随机分成两组,实验组12只小鼠均在背部用环钻制备圆形创面；而对照组6只小鼠不致伤。分别于1d、2d、3d、7d、14d、21d在实验组创缘和对照组小鼠背部相同部位切取全层皮肤组织标本,用抗鼠多克隆抗体P40进行抗△Np63免疫组化染色。结果：△Np63α在正常皮肤中的特征性表达模式是强阳性染色限于生发层的角质形成细胞核。伤后1～2d,表皮开始迁移,与对照组正常皮肤相比,迁移表皮舌基底层细胞△Np63α表达下调。伤后3d,△Np63α有较强的表达出现在创缘和迁移的表皮舌中,阳性染色限于基底层和其上方1～2层的角质形成细胞,而创面尚未被上皮覆盖处则没有阳性细胞。伤后5～7d,△Np63a有更强的表达出现在创缘和迁移表皮舌的基底层角质形成细胞。伤后14～21d创面愈合,△Np63α在覆盖创伤区表皮的基底层和棘层的细胞中高表达,并且其表达在表皮中长期和持续的存在。结论：△Np63α可能通过调控表皮修复中的细胞增殖和细胞迁移而参与正常皮肤的创面愈合过程。     

△Np63α在创面愈合过程中表达改变的研究

目的:探讨p63基因编码的蛋白亚型△Np63α在创面愈合表皮再生中的表达及其生物学意义.方法:①在3周龄小鼠背部用直径1.6cm的环钻压切制备出一个2.0cm2全层皮肤缺损的圆形创面,并将切除的皮肤原位缝合以覆盖创面.②将18只昆明小鼠随机分成两组,实验组12只小鼠均在背部用环钻制备圆形创面;而对照组6只小鼠不致伤.分别于1d、2d、3d、7d、14d、21d在实验组创缘和对照组小鼠背部相同部位切取全层皮肤组织标本,用抗鼠多克隆抗体P40进行抗△Np63免疫组化染色.结果:△Np63α在正常皮肤中的特征性表达模式是强阳性染色限于生发层的角质形成细胞核.伤后1～2d,表皮开始迁移,与对照组正常皮肤相比,迁移表皮舌基底层细胞△Np63α表达下调.伤后3d,△Np63α有较强的表达出现在创缘和迁移的表皮舌中,阳性染色限于基底层和其上方1～2层的角质形成细胞,而创面尚未被上皮覆盖处则没有阳性细胞.伤后5～7d,△Np63α有更强的表达出现在创缘和迁移表皮舌的基底层角质形成细胞.伤后14～21d创面愈合,△Np63α在覆盖创伤区表皮的基底层和棘层的细胞中高表达,并且其表达在表皮中长期和持续的存在.结论:△Np63α可能通过调控表皮修复中的细胞增殖和细胞迁移而参与正常皮肤的创面愈合过程.     

小鼠全层皮肤创伤愈合过程中基质细胞衍生因子-1及其受体CXCR4基因表达的研究

目的：研究小鼠全层皮肤创伤愈合过程中创面基质细胞衍生因子-1（stroma-cell derived factor-1,SDF-1）及其受体CXCR4的基因表达情况。方法：建立小鼠背部皮肤正中近颈侧1.5cm×1.5cm的正方形皮肤全层缺损模型,分别于伤后1、2、3、4、5、7、10和14d获取创缘组织,应用半定量逆转录-聚合酶链反应检测损伤后各时间点创面SDF-1及CXCR4的mRNA表达,并观察组织病理学变化。结果：伤后1～3d创面有大量炎症细胞浸润,4d时有肉芽组织形成,5d可见上皮细胞覆盖创面,7d时创面周围明显上皮化,14d创面基本完全愈合。SDF-1及CXCR4在创面愈合过程均呈双峰表达,SDF-1基因表达于伤后1d明显增高（P〈0.01）,随后下降,于伤后3d达最低,随后再次升高,于伤后5d达峰值（P〈0.01）,然后下降,14d时接近伤前值。CXCR4基因表达于伤后1d升高,然后继续升高,至伤后5d达峰值,随后下降,于伤后10d表达最少（P〈0.01）,伤后14d表达再次增加（P〈0.01）。结论：SDF-1/CXCR4轴参与了创伤愈合的炎症反应期和增殖期的愈合过程,在皮肤组织创伤愈合过程中起着重要作用。     

扁桃体剥离术后应用人表皮生长因子的疗效观察

目的：观察人表皮生长因子在全麻下扁桃体剥离术后应用对疼痛减轻及创面恢复的疗效。方法分析本院2008年10月至2010年3月全麻下扁桃体剥离术后病例132例，分为两组，治疗组72例术后予人表皮生长因子局部涂抹，1次／天，共3天，对照组60例术后6小时予朵贝氏液含漱，3次／天，共3天。结果比较两组的疼痛缓解时间及创面愈合时间，治疗组创面显效率为87．5％，疼痛消失率为72.2％：对照组创面愈合率为56．7％，疼痛消失率为33.3％，两组对比差异具有统计学意义（P〈O．05）。治疗组的疼痛缓解时间和创面愈合时间明显短于对照组。结论人表皮生长因予具有促进创面愈合功能，可减轻患者扁桃体剥离术后的痛苦，加快创面愈合，值得临床推广。     

肛肠病术后应用康复新液促进创面愈合的临床观察

为观察肛肠病术后应用康复新液促进创面愈合的临床疗效,将120例肛肠病术后患者随机分为治疗组和对照组,对照组采用常规换药,治疗组在常规处理基础上加用康复新液,比较两组创面愈合时间、创缘水肿差异。结果显示,治疗组创面愈合时间明显短于对照组．创面水肿临床发生率明显低于对照组。结果表明,康复新液对肛肠病术后创面有良好的促进愈合和降低水肿作用。     

仙方活命饮加减方治疗湿热毒蕴型低位肛周脓肿术后创面的临床疗效观察

目的：观察仙方活命饮加减方治疗湿热毒蕴型低位肛周脓肿术后创面的临床疗效。方法：选取2019年9月—2021年3月我院收治的低位肛周脓肿术后患者96例,分为对照组和试验组,每组48例。对照组采用常规治疗+温水熏洗、坐浴+外科换药,治疗组采用常规治疗+仙方活命饮加减方内服、熏洗坐浴+外科换药。比较两组患者炎症消退时间及创面愈合时间、创面疼痛、分泌物量及创缘水肿评分。结果：治疗组炎症消退时间为（6.67±1.69）d,创面愈合时间为(22.11±3.87) d;对照组炎症消退时间为(8.75±2.06) d,创面愈合时间为(26.55±3.93) d,两组创面炎症消退时间和愈合时间差异有统计学意义（P <0.01）。仙方活命饮加减方内服联合溻渍外用可减轻创缘水肿、减轻术后创面疼痛、减少创面炎性渗出,两组评分差异有统计学意义（P <0.05）。结论：治疗组经仙方活命饮加减方内服及溻渍外洗后创缘水肿消退快,渗出减少,疼痛减轻,肛门功能恢复快,创面愈合时间短。     

630～650nm光(红光)治疗烧伤创面的疗效观察

目的研究630~650nm可见光治疗烧伤创面的疗效及安全性评价。方法收集上海市第三人民医院烧伤整形科自2013年12月份至2014年12月份烧伤/创伤部位对称或临近的患者44例,共78个创面,随机分为治疗组和对照组。治疗组(n=40)创面使用630~650光治疗,对照组采用常规使用1%磺胺嘧啶银乳膏换药,两组共治疗21天,每日摄像,观察并计算创面愈合率、创面分泌物、创缘反应以及安全性评价。结果治疗组创面愈合率高于对照组,并减少创面分泌物、缓解创缘反应。结论临床观察表明,630~650nm光能减少创面分泌物的分泌,减轻创缘反应,缓解疼痛,促进创面愈合。     

表皮干细胞在创伤愈合中的作用

表皮干细胞（Epidermal stem cells,ESCs）是皮肤组织最重要的干细胞,具有多向分化潜能,在皮肤创伤愈合的各个阶段均发挥作用。了解ESCs在皮肤创伤愈合中的作用机制,将有助于临床皮肤创伤的修复,我们就ESCs在创伤愈合中作用的研究进展进行综述。     

Juglone ameliorates skin wound healing by promoting skin cell migration through Rac1/Cdc42/PAK pathway

Skin cell regeneration and wound healing are key processes in the recovery from skin injuries. Rapid cell migration and regeneration of skin cells lead to faster and better healing of wounded skin. In the present study, we aimed to investigate the wound healing potential of juglone, a naturally occurring Pin1 inhibitor found in walnuts. Cultured skin cells (NHDF and HaCaT) and hairless mice were treated with juglone after wound creation to examine its effects on cell migration and wound healing rate. The expressions of cell migration related proteins (Rac1, Cdc42, and α‐PAK), collagen deposition, and angiogenesis were analyzed. Juglone treatment resulted in faster rate of growth and migration and recovered cell morphology, particularly at a concentration of 5 µM, in skin cells compared to the untreated group. In vivo experiments showed that mice treated with juglone showed faster wound healing rate with better skin morphology and collagen deposition than the vehicle group. Furthermore, juglone increased the activation and/or expression of Cdc42, Rac1, and α‐pak in HaCaT cells, and resulted in enhanced angiogenesis in endothelial cells (HUVECs). Juglone also activated MAPKs signaling by activation of ERK, JNK, and p38 proteins. Taken together, these data suggest that juglone may be a potential candidate for wound healing and skin regeneration which ameliorates wound healing mainly by promoting skin cell migration through Rac1/Cdc42/PAK pathway.     

自体表皮细胞-纤维蛋白膜创面移植治疗大鼠烧伤

目的：观察自体表皮细胞-纤维蛋白膜移植到大鼠烧伤创面治疗皮肤缺损的效果。方法：健康Wistar大鼠20只,随机分成烧伤皮肤缺损造模组和自体表皮细胞-纤维蛋白膜移植治疗组,治疗后计算表皮细胞在纤维蛋白膜上最佳接种密度,观察移植后的各组创面愈合情况、创面伤口的收缩比例等。结果：在纤维蛋白膜上接种表皮细胞的最佳密度为5×10^4／cm2,烧伤皮肤缺损造模组创面完全愈合时间平均22．3d,自体表皮细胞-纤维蛋白膜移植治疗组为18．1d,造模组创面收缩率为（70±5）％,移植组为（20±5）％（均P〈0．05）。结论：自体表皮细胞-纤维蛋白膜可用于覆盖大面积烧伤造成的皮肤缺损,预防创面伤口瘢痕化的形成,减轻创面收缩率,加速皮肤缺损创面的愈合速度。     

Akermanite bioceramic enhances wound healing with accelerated reepithelialization by promoting proliferation,migration, and stemness of epidermal cells

Reepithelialization is an important step of wound healing, which is mainly completed by proliferation and migration of epidermal cells. Akermanite is a Ca‐, Mg‐, and Si‐containing bioceramic. This study evaluated the effects of Akermanite on wound healing and investigated the mechanisms. Using scald burn mice models, we demonstrated that local Akermanite treatment significantly accelerated wound healing by increasing reepithelialization and the stemness of epidermal cells. Epidermal cells were cultured in medium containing Akermanite extracts to explore the cellular mechanism of reepithelialization. Akermanite promoted the cell proliferation and migration, maintaining more cells in the S and G₂/M phases of the cell cycle. An additional study showed that Akermanite enhanced the expressions of integrinβ1, Lgr4, Lgr5, and Lgr6, which are specific molecular markers of epidermal stem cells, accompanied by the activation of the Wnt/β‐catenin pathway. These results suggested that Akermanite accelerated reepithelialization by increasing the proliferation, migration, and stemness of epidermal cells in a manner related to the Wnt/β‐catenin pathway, which might contribute, at least partially, to accelerated wound healing by Akermanite therapy.     

SCF increases in utero–labeled stem cells migration and improves wound healing

Diabetic skin wounds lack the ability to heal properly and constitute a major and significant complication of diabetes. Nontraumatic lower extremity amputations are the number one complication of diabetic skin wounds. The complexity of their pathophysiology requires an intervention at many levels to enhance healing and wound closure. Stem cells are a promising treatment for diabetic skin wounds as they have the ability to correct abnormal healing. Stem cell factor (SCF), a chemokine expressed in the skin, can induce stem cells migration, however the role of SCF in diabetic skin wound healing is still unknown. We hypothesize that SCF would correct the impairment and promote the healing of diabetic skin wounds. Our results show that SCF improved wound closure in diabetic mice and increased HIF‐1α and vascular endothelial growth factor (VEGF) expression levels in these wounds. SCF treatment also enhanced the migration of red fluorescent protein (RFP)‐labeled skin stem cells via in utero intra‐amniotic injection of lenti‐RFP at E8. Interestingly these RFP+ cells are present in the epidermis, stain negative for K15, and appear to be distinct from the already known hair follicle stem cells. These results demonstrate that SCF improves diabetic wound healing in part by increasing the recruitment of a unique stem cell population present in the skin.     

愈创膏联合干细胞移植治疗重度压疮的临床研究

目的探讨愈创膏联合干细胞移植治疗对重度压疮创面愈合及神经生长因子表达的影响。方法选取健康外伤患者30例手术中废弃的正常皮肤及皮下软组织作为对照组,将60例重度压疮患者随机分为两组各30例。观察组应用愈创膏联合庆大霉素纱布常规换药,治疗组应用愈创膏联合脐血干细胞移植治疗。比较治疗组与观察组愈合时间,同时测定三组创面神经生长因子(NGF)含量。结果治疗组创面愈合时间显著短于观察组(P<0.01)。观察组与治疗组治疗前NGF含量显著低于对照组(均P<0.01)。治疗后观察组与治疗组局部NGF含量显著升高,后期治疗6 d达到高峰,10 d逐步趋于正常,治疗组治疗后期3 d、6 dNGF含量显著高于观察组(均P<0.01)。结论愈创膏联合干细胞治疗可加速重度压疮创面的愈合,提高创面组织神经生长因子表达。     

Muscle-derived stem cells used to treat skin defects prevent wound contraction and expedite reepithelialization

Stem cells derived from adult tissues may serve as cell therapy to enhance the healing process in skin wounds. This study was designed to evaluate the use of autologous muscle-derived stem cells in an experimental skin wound model in terms of their efficiency at promoting tissue repair/regeneration. Muscle-derived cells obtained from the dorsal muscle of New Zealand rabbits were cultured in vitro for 2 weeks. The cell population was identified using the satellite markers CD34, m-cadherin and Myf5, and the proliferative capacity of the adult stem cells was determined. The population was then fluorescently labeled with PKH26 and seeded onto a circular 2 cm diameter defect created on the dorsal side of the ear of the rabbit from which the cells had been harvested. Similar defects on the contra lateral ears were left untreated to form the control group. Fourteen days later, specimens were taken for light, transmission, and scanning electron microscopy, as well as for immunolabeling with antibodies against vimentin, alpha-actin, desmin, myosin, fibronectin, and cytokeratin 14. Areas of wound contraction and reepithelialization were determined by image analysis. Wound contraction was significantly greater in the control than the treatment group (p<0.05); control specimens also showed more myosin expression. Reepithelialized areas were significantly greater in the treatment group (p<0.05). Control wounds showed nonepithelialized areas and inflammatory granulation tissue. Reepithelialization occurred as epidermal tongues of fusiform cells. Our findings indicate that the use of autologous stem cells on skin wounds expedites and improves the organism's natural healing process.     

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。     

三种不同免疫状态小鼠皮肤创面愈合过程中基质细胞衍生因子-1的表达

目的 观察3种不同免疫状态小鼠皮肤创面愈合过程中基质细胞衍生因子-1(SDF-1)的基因表达.方法 采用差异性贴壁法分离纯化BALB/C雄性小鼠骨髓源性干细胞(BMSCs),取第3代BMSCs进行成脂、成骨诱导鉴定;同时另取第3代BMSCs,经尾静脉注射到3.5 Gy放射性损伤后的雌性BALB/C鼠、裸鼠、SCID鼠体内,小鼠背部皮肤制作一直径为1.5 cm的创面,建立骨髓移植嵌合体皮肤创伤动物模型,观察创面的愈合过程,并于伤后1、3、5、7、14 d获取创面组织,应用半定量逆转录-聚合酶链反应(RT-PCR)检测创面组织中SDF-1的基因表达.结果 提取的BM-SCs可以成功进行成骨、成脂诱导分化;创面愈合过程中BALB/C鼠SDF-1基因表达于伤后1 d即有增高,5 d达峰值(P<0.01),然后逐渐下降,14 d创面基本愈合时接近对照组.裸鼠、SCID鼠创面中SDF-1基因表达于伤后逐渐增高,7 d达峰值(P<0.01),与BALB/C鼠比较,SDF-1基因表达峰值延后,14 d创面未愈,SDF-1基因表达仍高于对照组.结论 皮肤创面愈合过程中,SDF-1趋化BMSCs,参与创面愈合的炎症反应期和增殖期的修复过程,在创面愈合中起着重要作用;机体的免疫功能也将影响BMSCs募集,从而影响创面愈合.