点阵CO2激光作用小鼠皮肤光老化模型后创面愈合过程中皮肤超微结构的变化

目的:观察点阵CO2激光作用小鼠皮肤光老化模型后创面愈合过程表皮的超微结构的变化规律。方法:将5只昆明小鼠以UVB紫外线照射,制备成皮肤光老化模型,行点阵CO2激光(Deep FX)干预,观察激光创面愈合情况,并且分别于干预前、干预后第1、3、7、15天取材,进行HE染色和电镜观察。结果:点阵CO2激光作用于小鼠皮肤光老化模型后,术后第1天,见激光损伤灶周围变性组织及细胞水肿,成纤维细胞线粒体体积增大;术后第3天见表皮细胞向创底迁移,成纤维细胞出现增生,细胞内线粒体增多,粗面内质网扩张,胶原纤维见周期性横纹;术后第7天激光损伤灶创底见表皮细胞增殖,创面基本愈合,成纤维细胞增生活跃,线粒体、粗面内质网丰富,内质网扩张明显、胶原纤维增多增粗;术后第15天,成纤维细胞趋于稳定,胶原纤维排列密集。结论:点阵CO2激光通过对表皮的线粒体、粗面内质网产生影响,达到对光老化皮肤良好疗效。     

脂肪干细胞来源外泌体对人脐静脉血管内皮细胞增殖、迁移及管样分化的影响

目的探讨脂肪干细胞来源外泌体(adipose-derived stem cell released exosomes,ADSC-Exos)对人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖、迁移及管样分化的影响。方法取吸脂患者自愿捐赠的脂肪组织,采用酶消化法分离培养获得ADSCs,并行流式细胞检测及成脂诱导鉴定。收集第3代ADSCs上清液提取外泌体,透射电镜观察其形态,Western blot检测其膜表面标志性蛋白Alix和CD63,用纳米颗粒跟踪分析仪NanoSight分析外泌体粒径分布范围。用PKH26荧光标记的ADSC-Exos与HUVECs共培养后,共聚焦显微镜观察ADSC-Exos能否进入HUVECs。将ADSC-Exos与HUVECs共培养1、2、3、4、5 d,CCK-8法检测ADSC-Exos对HUVECs增殖影响;共培养12 h时ELISA法检测细胞上清液VEGF蛋白表达量。采用Transwell迁移实验检测ADSC-Exos对HUVECs迁移能力的影响。通过体外Matrigel基质胶实验,观察ADSCExos对HUVECs管状结构形成的影响;将HUVECs与Matrigel基质胶混合后联合ADSC-Exos注射在BALB/c雄性裸鼠皮下,2周后检测基质胶中新生血管情况,计算平均血管密度(mean blood vessel density,MVD)。上述实验均以等量PBS作为对照。结果经鉴定所培养细胞符合ADSCs的特征。ADSC-Exos为形态一致的圆形或椭圆形膜性囊泡,表达标志性蛋白Alix和CD63,粒径范围30~200 nm。共聚焦显微镜观察示ADSC-Exos可被HUVECs摄取。CCK-8法检测示,各时间点实验组细胞增殖均优于对照组(P0.05)。Transwell实验结果显示,实验组跨膜迁移细胞数较对照组显著增多(t=9.534,P=0.000)。在体外Matrigel胶成管实验中,实验组管样结构数明显多于对照组(t=15.910,P=0.000);在体内实验中,实验组MVD亦显著高于对照组(t=16.710,P=0.000)。ELISA检测示,实验组细胞上清液VEGF蛋白表达量明显高于对照组(t=21.470,P=0.000)。结论 ADSC-Exos可促进HUVECs增殖、迁移及管样结构形成,提示ADSC-Exos可促进血管新生。     

脂肪干细胞来源外泌体对人脐静脉血管内皮细胞增殖、迁移及管样分化的影响北大核心CSCD

目的探讨脂肪干细胞来源外泌体(adipose-derived stem cell released exosomes,ADSC-Exos)对人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖、迁移及管样分化的影响。方法取吸脂患者自愿捐赠的脂肪组织,采用酶消化法分离培养获得ADSCs,并行流式细胞检测及成脂诱导鉴定。收集第3代ADSCs上清液提取外泌体,透射电镜观察其形态,Western blot检测其膜表面标志性蛋白Alix和CD63,用纳米颗粒跟踪分析仪NanoSight分析外泌体粒径分布范围。用PKH26荧光标记的ADSC-Exos与HUVECs共培养后,共聚焦显微镜观察ADSC-Exos能否进入HUVECs。将ADSC-Exos与HUVECs共培养1、2、3、4、5 d,CCK-8法检测ADSC-Exos对HUVECs增殖影响;共培养12 h时ELISA法检测细胞上清液VEGF蛋白表达量。采用Transwell迁移实验检测ADSC-Exos对HUVECs迁移能力的影响。通过体外Matrigel基质胶实验,观察ADSCExos对HUVECs管状结构形成的影响;将HUVECs与Matrigel基质胶混合后联合ADSC-Exos注射在BALB/c雄性裸鼠皮下,2周后检测基质胶中新生血管情况,计算平均血管密度(mean blood vessel density,MVD)。上述实验均以等量PBS作为对照。结果经鉴定所培养细胞符合ADSCs的特征。ADSC-Exos为形态一致的圆形或椭圆形膜性囊泡,表达标志性蛋白Alix和CD63,粒径范围30~200 nm。共聚焦显微镜观察示ADSC-Exos可被HUVECs摄取。CCK-8法检测示,各时间点实验组细胞增殖均优于对照组(P<0.05)。Transwell实验结果显示,实验组跨膜迁移细胞数较对照组显著增多(t=9.534,P=0.000)。在体外Matrigel胶成管实验中,实验组管样结构数明显多于对照组(t=15.910,P=0.000);在体内实验中,实验组MVD亦显著高于对照组(t=16.710,P=0.000)。ELISA检测示,实验组细胞上清液VEGF蛋白表达量明显高于对照组(t=21.470,P=0.000)。结论 ADSC-Exos可促进HUVECs增殖、迁移及管样结构形成,提示ADSC-Exos可促进血管新生。     

肝素对大鼠Ⅱ度烧伤创面愈合的影响

目的　研究肝素对大鼠 度烧伤创面愈合的影响。方法　 2 0只 SD大鼠随机分为两组 ,制成2 0 %面积深 度烧伤 ,实验组采用肝素 10 0 U /kg加生理盐水至 1ml皮下注射 ;对照组采用生理盐水 1ml皮下注射 ,每天一次直至创面完全愈合。从大体观察创面愈合天数及全身有无出血现象 ,光镜观察肉芽组织及胶原纤维生长情况 ,电镜观察成纤维细胞生长情况。结果　大鼠全部成活。实验组创面愈合时间为 (2 2 .8± 1.87)天 ,对照组为(2 6 .2± 2 .82 )天 ,实验组创面愈合所需时间明显少于对照组 (P<0 .0 0 5 )。光镜观察发现实验组肉芽组织及胶原纤维生长明显优于对照组。电镜观察发现实验组成纤维细胞生长优于对照组。结论　肝素皮下注射能加速创面愈合。     

人脐带MSCs-海藻酸盐凝胶敷料的实验研究

目的观察人脐带MSCs(human umbilical cord MSCs,hUCMSCs)在海藻酸盐凝胶支架中的生长特性,探讨hUCMSCs-海藻酸盐敷料修复创面的可行性。方法取人脐带标本体外分离培养hUCMSCs,取第4代细胞与海藻酸钙凝胶支架复合培养(实验组)。倒置相差显微镜下观察细胞在凝胶支架中的生长特征,培养第0、3、6、9天检测细胞上清液VEGF含量并行细胞计数,并检测细胞迁移能力;以正常培养hUCMSCs作为对照组。取32只8周龄Balb/c雄性小鼠,制备全层皮肤缺损模型,随机分为4组(n=8);创面分别以hUCMSCs-海藻酸钙凝胶复合物(MSC-凝胶组)、细胞上清液-海藻酸钙凝胶复合物(CS-凝胶组)、10%FBS-海藻酸钙凝胶复合物(FBS-凝胶组)及0.01 mol/L PBS-海藻酸钙凝胶复合物(PBS-凝胶组)修复。观察创面愈合情况,计算第5、10、15天创面愈合率;于第15天切取创面组织行组织学及免疫组织化学染色,评估新生皮肤情况。结果 hUCMSCs在凝胶支架中可生长增殖达1周以上,形似葡萄样;细胞迁移实验示7 d内未见细胞从凝胶支架中迁移出。培养第3天实验组细胞数及VEGF表达量均明显少于对照组(P0.05),之后逐渐增加,至第9天时均明显多于对照组(P0.05)。动物实验显示,与FBS-凝胶组及PBS-凝胶组相比,MSC-凝胶组及CS-凝胶组第5、10、15天创面愈合率均显著增高(P0.05),新生皮肤鳞状上皮、成纤维细胞、皮脂腺及毛细血管均增多,第15天时VEGF表达量也显著增加(P0.05);而MSC-凝胶组及CS-凝胶组观察情况相似。结论 hUCMSCs能在海藻酸钙凝胶中持续表达创面愈合所必需的VEGF,hUCMSCs-海藻酸钙凝胶复合物能促进小鼠创面愈合,有望成为理想的创面敷料。     

碱性成纤维细胞生长因子对大鼠皮瓣愈合的实验研究

目的 观察碱性成纤维细胞生长因子(bFGF)对大鼠皮瓣损伤后创面愈合及血管生成的作用.方法 Wistar大鼠24只,制作大鼠皮瓣模型.实验组创面使用bFGF在皮瓣距离蒂部2、5、8 cm处分别注射bFGF 100 U(约0.025 ml)于皮瓣基底部,于第1、5、10天各重复1次,共3次,对照组创面给予等量生理盐水作为对照,观察其创面愈合时间及血管的变化,并分别取相同部位皮瓣组织块行病理组织学检查及免疫组化分析.结果 实验组大鼠皮瓣创面愈合时间比对照组缩短(3.0±0.56)d,创面新生血管内皮生长因子阳性表达增多,新鲜肉芽组织形成增多.结论 碱性成纤维细胞生长因子有促进大鼠皮瓣损伤后血管内皮生长因子的表达,刺激内皮细胞分裂,促进新生血管及肉芽组织形成,从而加速皮瓣损伤后愈合.     

Ⅲ度烧伤后肉芽切除断层皮下组织创面自体皮移植对创面愈合的影响

目的探讨对Ⅲ度烧伤后肉芽切除断层皮下组织创面,采用自体皮移植修复的机制,并评价其疗效.方法采用肉眼、病理组织学、免疫组织化学、电镜和流式细胞术,动态观察成年Ⅲ系小型猪Ⅲ度烧伤后肉芽切除断层皮下组织创面自体植皮术(A组)后,移植皮片活性与结构,移植床肉芽组织、胶原纤维、微血管、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)蛋白表达和成纤维细胞超微结构变化,并与Ⅲ度烧伤后传统的肉芽刮除纤维板创面自体植皮术(B组)进行比较,另设对照组,皮肤仅剃毛(C组). 结果术后3天,移植皮片成活,A组较B组皮片损伤轻、皮肤细胞增殖指数高、移植床血管内皮细胞及肉芽组织增生,bFGF表达明显.5天,两组血管内皮细胞及肉芽组织增殖旺盛,成纤维细胞呈蛋白合成旺盛和功能活跃象,新生胶原纤维出现,但B组增殖更加明显.7～14天,A组表皮结构及真皮毛细血管密度逐渐恢复正常,移植床肉芽组织逐渐成熟为纤维结缔组织,而B组则延后2天左右.21天后,A组愈合创面病理改变轻于B组.30～60天,A组创面收缩及改容程度明显轻于B组,且触之较软、移动度尚可. 结论Ⅲ度烧伤后肉芽切除断层皮下组织创面自体皮移植术的疗效优于传统手术.     

盐酸普萘洛尔乳膏对糖尿病小鼠创面愈合的影响及其机制研究

目的探讨局部外用盐酸普萘洛尔乳膏对糖尿病小鼠创面愈合的影响及其可能机制。方法取8周龄雌性自发性BKS.Cg-Dock7m+/+Leprdb/JNju糖尿病小鼠18只,随机分为实验组及对照组(n=9)。于两组小鼠背部左、右侧各制备直径为0.6 cm的圆形全层皮肤缺损创面,实验组及对照组创面分别涂抹含或不含盐酸普萘洛尔的乳膏,术后2、5、7、10、14、17 d重复涂抹1次。术后2、5、7、10、14、17、21 d,观察创面愈合情况,计算创面愈合率;取材行HE染色、Masson染色、甲苯胺蓝染色,观察创面再上皮化、胶原纤维、肥大细胞情况,Western blot检测创面组织中IL-1β和促血管生成素2(angiogenin 2,Ang2)蛋白的表达。结果两组创面均逐渐愈合,但实验组创面愈合速度快于对照组,除术后21 d外,其余各时间点实验组创面愈合率均高于对照组,比较差异有统计学意义(P0.05)。组织学观察示,与对照组相比,实验组创面再上皮化加快,创面内肥大细胞分布较少。Western blot检测示除术后10 d外,其余各时间点实验组IL-1β蛋白相对表达量均低于对照组;术后各时间点实验组Ang2蛋白相对表达量均低于对照组。结论局部外用盐酸普萘洛尔乳膏可促进自发性糖尿病小鼠创面愈合,可能与其减少炎性反应、稳定血管及促进表皮细胞增殖等有关。     

猪内脏膜生物敷料促进兔皮肤创面愈合的实验研究

目的 观察一种用猪内脏膜制成的新型生物敷料对兔皮肤创面愈合的影响.方法 制做兔背全层创伤模型,分成实验组和对照组,创面分别外用新型生物敷料及无菌凡士林敷料.伤后3、5、7、10、14、17、21 d观察两组创面愈合情况和创面愈合率;并分别取创面组织进行病理组织学、羟脯氨酸含量测定及成纤维细胞周期分析等检查,评估创面的修复质量,进行观察比较.两组数据的比较采用t检验,两组间组织病理结构评价采用秩和检验,P＜0.05为差异有统计学意义.结果 实验组术后7、10、14、17、21 d创面愈合率分别比对照组高,差异具有统计学意义(P＜0.05);术后第14天进行两组间组织病理等级评价,发现实验组在组织结构、表皮层、基底细胞层及真皮层评价中等级占+++的百分比分别比对照组高.两组之间对比采用秩和检验,差异具有统计学意义(P＜0.05);实验组与对照组平均愈合时间分别为(12.36±2.03)d、(16.82±2.12)d,差异有统计学意义(P＜0.05);实验组在3、5、7、10、14、17及21 d时,组织羟脯氨酸的含量分别比对照组高,差异有统计学意义(P＜0.05);创面组织成纤维细胞细胞周期分析结果显示:实验组每时相S期细胞百分比均高于对照组.结论 用猪内脏膜制成的新型生物敷料可加速皮肤创面上皮化过程,使伤口愈合时间提前.可提高皮肤创面愈合质量,促进胶原生成的作用.     

可溶性CD100蛋白促进糖尿病小鼠创面愈合的实验研究

目的研究外源性CD100分子对糖尿病小鼠创面愈合的影响。方法在BKS糖尿病小鼠背部制备直径5 mm的圆形创面,术后当天CD100组在每个创面局部皮下注射CD100 250 ng(50滋l),对照组注射PBS,两组均隔天注射直至创面完全愈合。术后隔天拍照,计算创面面积占原始面积比率。HE和Masson染色行组织学检查,CD34免疫组化染色并标记新生血管,CD68染色并标记巨噬细胞。结果由于血痂的影响,CD100组与PBS组相比创面面积无显著差异。组织学结果表明,第7、13、21天CD100组新生表皮与真皮的新生评分显著优于PBS组(P0.05);肉芽组织厚度评分在第7、13天大于PBS组,第21天小于PBS组(P0.05),胶原重塑优于PBS组。在各个时间点CD100组血管化程度优于PBS组(P0.05);炎症状态轻于PBS组(P0.05)。结论局部应用外源性CD100分子,能够通过促进创面的血管化,减轻炎症反应促进糖尿病小鼠创面愈合。     

透明质酸外敷膜对大鼠手术切口愈合

目的　探讨透明质酸外敷膜对手术切口愈合的影响。方法　选用纯系SD大鼠 4 8只 ,随机分为 8组 ,每组 6只。采用背部两侧切口 ,左侧外敷透明质酸膜 ,为实验组 ;右侧外敷生理盐水纱条作为对照。分别对术后 1～ 2 8d切口愈合情况进行组织学观察。结果　实验组炎症反应轻 ,表皮愈合快 ,成纤维细胞发生退行样变化 ,胶原纤维较纤细 ,排列较疏松。结论　透明质酸外敷膜在切口愈合早期 ,有较强的透皮作用 ;并可促进表皮层愈合、延缓角化层形成。     

Delayed cutaneous wound healing in aged rats compared to younger ones

Delayed wound healing in elderly males is a complex process in which the factors responsible are not fully understood. This study investigated the hormonal, oxidative and angiogenic factors affecting wound healing in aged rats. Two groups consisting of eight healthy male Wistar Albino rats [young (30 ± 7 days) and aged (360 ± 30 days)], and a cutaneous incision wound healing model were used. Scar tissue samples from wounds on the 7th, 14th and 21st days of healing were evaluated for hydroxyproline and vascular endothelial growth factor content. Macrophage, lymphocyte, fibroblast and polymorphonuclear cell infiltration; collagen formation and vascularization were assessed by light and electron microscopy. The free oxygen radical content of the wounds was measured by a chemiluminescence method. Blood sample analysis showed that the hydroxyproline and total testosterone levels were significantly higher, and the oxygen radical content was significantly lower in young rats. Histopathological, immunohistochemical and ultrastructural evaluations revealed higher amounts of fibroblasts and collagen fibers, and more vascularization in young rats. These results are indicative of the delayed wound healing in aged rats. A combination of multiple factors including hormonal regulation, free oxygen radicals and impaired angiogenesis appears to be the cause of delayed cutaneous healing.     

Comparison of the effects of topical fusidic acid and rifamycin on wound healing in rats

Wound healing is an active and dynamic process that begins from the moment of injury. Any delay in the initiation of the response to injury can prolong the healing process. The aim of this study was to investigate the effects of topically applied fusidic acid and rifamycin on wound healing in a full‐thickness wound model. Ten female Sprague‐Dawley rats, aged 4 months and weighing 200–250 g, were used. Four rifamycin (R), four fusidic acid (F) and four control (K) areas were generated on their backs by using a 5‐mm punch biopsy pen. On the 4th, 7th, 14th and 21st days, biopsies were taken from each wound area of all the rats. Fusidic acid group demonstrated a statistically significant increase of collagen and intensity of fibroblast proliferation on the 21st day of wound healing, whereas in the rifamycin group, healing time was, as expected, similar to physiological wound‐healing phases. Despite the limited number of subjects, topical fusidic acid was found to delay wound healing by prolonging fibroblast proliferation.     

Raman spectroscopy enables noninvasive biochemical identification of the collagen regeneration in cutaneous wound healing of diabetic mice treated with MSCs

Mesenchymal stem cells (MSCs) had been reported as a novel therapeutic strategy for non-healing diabetic cutaneous wound mainly by promoting the formation of extracellular matrix (ECM) and neovasculature. Collagen regeneration is one of the key processes of ECM remodeling in wound healing. Accordingly, rapid assessment of the collagen content in a noninvasive manner can promptly provide objective evaluation for MSC therapy of cutaneous wound healing and strength evidence to adjust therapeutic regimen. In the present study, noninvasive Raman microspectroscopy was used for tracing the regeneration status of collagen during diabetic wound healing with MSCs. Wound tissues of normal mice, diabetic mice, and MSC-treated diabetic mice were subjected to Masson trichrome staining assay and submitted to spectroscopic analysis by Raman microspectroscopy after wounding 7, 14, and 21 days. Masson trichrome staining demonstrated that there was more collagen deposition in diabetic + MSCs group relative to diabetic group. The relative intensity of Raman collagen peak positions at 937, 1004, 1321, 1452, and 1662 cm^?1 increased in MSC-treated diabetic group compared to diabetic group, although normal mice group had the highest relative intensity of collagen peak bands. Correlation analysis suggested that the spectral bands had a high positive correlation with the collagen intensity detected by Masson trichrome staining in wound tissues of three groups. Our results demonstrate that Raman microspectroscopy has potential application in rapidly and quantitatively assessing diabetic wound healing with MSCs by monitoring collagen variation, which may provide a novel method for the study of skin regeneration.     

体外震波治疗对糖尿病大鼠慢性创面愈合及组织学影响

目的体外震波(extracorporeal shock wave,ESW)可促进血管新生和组织修复,通过观察低能量ESW治疗糖尿病大鼠慢性创面的效果,探讨其促进糖尿病慢性创面愈合的机制。方法 6～8周龄雄性SD大鼠96只,体重(220±20)g,随机分为3组(糖尿病对照组、ESW治疗组及正常对照组),每组32只。取糖尿病对照组、ESW治疗组大鼠,腹腔注射链脲佐菌素(60 mg/kg)制备糖尿病大鼠模型后,于背部制备1个直径1.8 cm的圆形皮肤全层创面,建立糖尿病大鼠慢性创面模型。正常对照组大鼠同法制备创面。创面制备后1 d ESW治疗组采用0.11 mJ/mm2、1.5 Hz的ESW对创面干预500脉冲;糖尿病对照组及正常对照组创面未行ESW治疗。观察创面愈合情况;于3、7、14 d取创面组织行HE及Masson染色,观察创面组织学变化;行CD31和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)免疫组织化学染色,观察创面细胞增殖及血管化程度。结果糖尿病对照组创面闭合率低于正常对照组,创面闭合时间延长(P<0.05),治疗后3、7、14 d,创面组织炎性细胞浸润明显,胶原纤维相对面积密度、创面微血管密度、PCNA阳性细胞相对密度均低于正常对照组(P<0.05)。ESW治疗组与糖尿病对照组相比,创面愈合时间缩短,创面闭合率提高(P<0.05),各时间点炎性细胞计数减少,肉芽组织生长良好,创面内胶原纤维相对面积密度、微血管密度和PCNA阳性细胞相对密度增加,差异有统计学意义(P<0.05);ESW治疗组与正常对照组相比,微血管密度和PCNA阳性细胞相对密度差异均无统计学意义(P>0.05)。结论低能量ESW治疗可以抑制糖尿病大鼠慢性创面内局部炎性反应,提高创面内修复细胞增殖能力,增加微血管形成和胶原沉积,使肉芽组织形成增加,最终促进创面愈合。     

生肌长皮膏促进下肢慢性溃疡愈合的病例对照研究

目的:探讨中药生肌长皮膏促进下肢慢性溃疡愈合的临床疗效。方法:2010年3月至2011年9月外科门诊和中医科收治的64例患有下肢慢性溃疡病人随机分为实验组和对照组,每组32例。实验组溃疡创面应用中药生肌长皮膏,对照组采用普通换药治疗,均为每天1次。两组病人于治疗后第3、7、14和21 d观察并计算创面愈合率,并分别取创面肉芽组织标本行HE染色、Masson染色和免疫组织化学检测CD34,观察组织结构变化、总胶原的分布和微血管密度(MVD)。结果:与对照组相比,实验组在第14、21 d创面愈合率明显较高;胶原着色深、面积广[(68.7±7.7)比(48.9±9.3),P<0.05)。结论:中药生肌长皮膏可促进创面肉芽组织生长、胶原合成和血管新生,从而促进溃疡愈合,有益于治疗下肢慢性溃疡。     

组织工程口腔黏膜固有层修复皮肤缺损的初步研究

目的探讨两种组织工程口腔黏膜固有层移植修复皮肤缺损的效果.方法分别以胶原凝胶和壳多糖-胶原凝胶为网架与体外培养的Wistar大鼠口腔黏膜成纤维细胞构建胶原凝胶口腔黏膜固有层(fibroblast-populated collagen lattice,FPCL)、壳多糖-胶原凝胶口腔黏膜固有层(fibroblast -populated chitosan collagen lattice,FPCCL),用BrdU标记其中的成纤维细胞后,移植修复同种异体大鼠背部全层皮肤圆形缺损.将36只21～25周龄Wistar大鼠分为FPCL组、FPCCL组及创口仅覆盖纱布的对照组,每组12只.术后行大体观察创面愈合情况;4、7、14和21 d 3组创面直径测量;组织学及免疫组织化学染色观察其组织修复情况.结果术后大鼠创面均无感染,创面结痂在14及17 d自然脱落,创面逐渐愈合,被新生表皮覆盖,术后21 d新生皮肤光滑,颜色与正常皮肤接近,无体毛.术后各时间点3组创面直径均逐渐变小,差异有统计学意义(P＜0.01),14 d以后,创口大小较稳定.术后7 d,FPCL组受植创面比FPCCL组受植创面和对照组创面小(P＜0.01).组织学观察:术后7 d,3组创面未完全表皮化;14、21 d,FPCL组、FPCCL组受植区完全表皮化,有钉突,对照组无明显钉突,表皮已分层,基底细胞层完整,最表面有角化物;真皮中新生胶原纤维细,含毛细血管.免疫组织化学染色显示,FPCL组、FPCCL组术后各时间点阳性成纤维细胞出现在肉芽组织的细胞密集处和新生真皮中,与新生肉芽组织共同参与了皮肤缺损的修复重建,未出现免疫排斥现象.结论口腔黏膜成纤维细胞作为修复皮肤缺损的种子细胞是可行和有效的,壳多糖胶原凝胶在限制受植区创面收缩变小方面优于单纯胶原凝胶.FPCL和FPCCL两组新生皮肤的质量优于对照组,两种组织工程口腔黏膜固有层作为皮肤缺损区永久性真皮替代物是可行和有效的.     

京万红软膏治疗糖尿病慢性创面的实验研究

目的：探讨京万红软膏促进糖尿病小鼠慢性创面愈合的效果及机制.方法：清洁级、同周龄雄性C57小鼠54只,随机分为京万红软膏组、对照药物组和空白对照组,每组18只.采用连续3 d腹腔注射1%链脲佐菌素60 mg/kg的方法建立糖尿病小鼠模型,然后在小鼠背部制作直径1 cm的圆形全层皮肤缺损创面.创面形成第2日开始,京万红软膏组创面涂抹京万红软膏,厚度0.5 cm; 对照药物组创面涂抹相同厚度复方磺胺嘧啶锌凝胶剂;空白对照组不给药.3组创面均以凡士林纱布覆盖,每日换药1次.于给药后第3、7、10、17、21天,用塑料透明膜描记创面大小,计算创面愈合率.于给药后第7、17天各组分别随机选取3只小鼠处死,观察创面及创缘2 mm范围内全层皮肤及肉芽组织的大体形态变化及组织病理学变化.结果：给药后7 d,两药物治疗组创面愈合率均有所升高,但3组间差异无显著性（P〉0.05）.给药后10、17和21 d,对照药物组创面愈合率[（81.00±0.85）%,（95.00±0.29）%,（97.00±0.37）% ]明显高于空白对照组[（77.00±1.35）%,（87.00±1.17）%,（90.00±0.96）%,P〈0.05],而京万红软膏组愈合率[（85.00±1.93）%,（100.00±0）%,（100.00±0）% ]明显高于对照药物组（P〈0.05）.治疗17 d京万红软膏组创面全部愈合,而对照药物组和空白对照组在治疗后21 d仍未全部愈合.组织病理学观察发现,京万红软膏较对照药物复方磺胺嘧啶锌凝胶剂能更早使炎症减轻、创面上皮化,而且表皮细胞复层分化良好,新生胶原排列整齐,创面组织结构恢复正常化明显,显示具有更好的愈合质量.结论：京万红软膏对于糖尿病慢性创面有良好的促进愈合作用.     

Lentiviral gene therapy with platelet-derived growth factor B sustains accelerated healing of diabetic wounds over time

The treatment of diabetic wounds is a formidable clinical challenge. In this study, lentiviral vectors carrying the human platelet-derived growth factor B (PDGF-B) gene were used to treated diabetic mouse wounds. Full-thickness 2.0-cm x 2.0-cm excisional wounds were created on the dorsa of genetically diabetic C57BL/KsJ-m+/+Lepr(db) mice. Lentiviral vectors containing the PDGF-B gene were injected into the wound margins and base. Mice were killed at 14-, 21-, and 35-day intervals. Measurement of the residual epithelial gap showed a trend towards increased healing in lentiviral PDGF-treated wounds compared with untreated and saline-treated wounds at all time points. At 21 days, there was significantly increased healing in lentiviral PDGF-treated wounds (0.98+/-0.17 cm) compared with saline-treated wounds (1.22+/-0.30 cm; P<0.05). Immunohistochemistry for CD31 revealed significantly increased neovascularization in lentiviral PDGF-treated wounds compared with untreated and saline-treated wounds at 14 and 21 days (P<0.01). Picrosirius red staining demonstrated thicker and more highly organized collagen fibers in treated wounds compared with untreated and saline-treated wounds. Quantitative analysis of collagen content showed a 3.5-fold and 2.3-fold increase in lentiviral PDGF-treated wounds versus untreated and saline-treated wounds, respectively (P<0.01). Lentiviral gene therapy with PDGF-B can sustain diabetic wound healing over time and may possess promising potential in the clinical setting.     

Evaluation of a new type of wound dressing made from recombinant spider silk protein using rat models

This study investigates the feasibility of recombinant spider silk protein as a wound-dressing material for coverage of deep second-degree burn wounds using an animal model. Sixty Sprague–Dawley (SD) rats were randomly divided into four groups (15 rats in each group). Two types of recombinant spider silk proteins, pNSR-16 and pNSR-32, as well as collagen (as a control) were applied on the wound; the fourth group was left untreated as a negative control. Each group was evaluated on the 3rd, 5th, 7th, 14th and 21st days for wound-healing rate, histological test, levels of hydroxyproline synthesis and the samples were stained for immunohistochemical detection of the basic fibroblast growth factor (bFGF). The results of implantation testing showed that wound healing in the treatment groups – recombinant spider silk protein pNSR-16 and pNSR-32 – was much better than that in the control group (p < 0.01). On the 7th, 14th and 21st days, higher expression of bFGF and the increase of hydroxyproline content of the skin indicated good regeneration of wound skin in the treatment groups. Preliminarily, we conclude that the recombinant spider silk protein membrane promotes the recovery of wound skin by increasing the expression and secretion of the growth factor bFGF and hydroxyproline.