大鼠烫伤创面感染模型的研制

Objective To reproduce a reliable rat model of burn with infection for the study of prevention and treatment of infected wound. Methods ( 1 ) Electrical burn producing apparatus equipped with constant temperature (80 ℃ ) and pressure (0.5 kg) was used to reproduce burn injury (with area of 4.5 cm2 ) on both sides of the back in 50 SD rats for different duration (4, 6, 8, 10, 12 s) , with 10 rats for each burn duration. On post burn day (PBD) 1, gross condition of wounds was observed with naked eyes.Wounds on the left side were used to observe healing time. The wounds on the right side were used for histological observation to determine the depth of injury, and they were classified into superficial and deep partialthickness injury. (2) Another 36 SD rats were divided into A (inflicted with superficial partial-thickness burn, n = 18) and B (inflicted with deep partial-thickness burn, n = 18) groups according to the random number table. Rats in both groups were treated in accordance with method of preliminary experiment. Immediately after burn, 0. 1 mL of liquid containing 1 × 109, 1 × 107, 1 × 105 CFU Pseudomonas aeruginosa (PA) ATCC 27853 was respectively inoculated to the wounds on one side (with 6 rats for each amount) ,while the wounds on the other side were treated with the same volume of normal saline as control. Inflammatory reaction of wounds was examined with HE staining on post inoculation day (PID) 1. On PID 1, 2, 3,5, 7, and 14, the number of subeschar bacteria was respectively counted and the bacteria were identified with Gram stain and biochemical reaction. Wound healing time was recorded. Data were processed with t test. Results (1) Burn for 6, 8 s was respectively identified as injury time resulting in superficial or deep partial-thickness injury according to histological observation and wound healing time. (2) Obvious inflammatory cell infiltration was observed in the wounds in B group which were inoculated with 1 × 107 , 1 ×109 CFU PA, and the infiltration was less marked in A group with inoculation of 1 × 109 CFU PA. (3) The bacteria isolated from wounds of A and B groups was identified as PA. The subeschar bacteria count within PID 14 in A group, in which different amount of PA was inoculated, was mostly less than 1 × 105 CFU/g of tissue, while that in B group in which 1 × 109 CFU PA was inoculated was more than 1 × 105 CFU/g of tissue. (4) There was no obvious difference in wound healing time between wounds inoculated with different amount of PA and wounds treated with normal saline in A group ( with t value respectively 1.26, 0. 29, 1.07,P values all above 0.05 ). Wound healing time of wounds in B group, in which 1 × 109 CFU PA was inoculated, was longer as compared with that treated with normal saline [(22.5 + 1.0) d vs. ( 19.4 + 1.6) d, t =2.73, P ＜0. 05]. Conclusions In rat, deep partial-thickness burn wound inoculated with 1 × 109 CFU PA ATCC 27853 is a reliable model with high reproducibility for the study of infection of burn wound.     

大鼠烫伤创面感染模型的研制

Objective To reproduce a reliable rat model of burn with infection for the study of prevention and treatment of infected wound. Methods ( 1 ) Electrical burn producing apparatus equipped with constant temperature (80 ℃ ) and pressure (0.5 kg) was used to reproduce burn injury (with area of 4.5 cm2 ) on both sides of the back in 50 SD rats for different duration (4, 6, 8, 10, 12 s) , with 10 rats for each burn duration. On post burn day (PBD) 1, gross condition of wounds was observed with naked eyes.Wounds on the left side were used to observe healing time. The wounds on the right side were used for histological observation to determine the depth of injury, and they were classified into superficial and deep partialthickness injury. (2) Another 36 SD rats were divided into A (inflicted with superficial partial-thickness burn, n = 18) and B (inflicted with deep partial-thickness burn, n = 18) groups according to the random number table. Rats in both groups were treated in accordance with method of preliminary experiment. Immediately after burn, 0. 1 mL of liquid containing 1 × 109, 1 × 107, 1 × 105 CFU Pseudomonas aeruginosa (PA) ATCC 27853 was respectively inoculated to the wounds on one side (with 6 rats for each amount) ,while the wounds on the other side were treated with the same volume of normal saline as control. Inflammatory reaction of wounds was examined with HE staining on post inoculation day (PID) 1. On PID 1, 2, 3,5, 7, and 14, the number of subeschar bacteria was respectively counted and the bacteria were identified with Gram stain and biochemical reaction. Wound healing time was recorded. Data were processed with t test. Results (1) Burn for 6, 8 s was respectively identified as injury time resulting in superficial or deep partial-thickness injury according to histological observation and wound healing time. (2) Obvious inflammatory cell infiltration was observed in the wounds in B group which were inoculated with 1 × 107 , 1 ×109 CFU PA, and the infiltration was less marked in A group with inoculation of 1 × 109 CFU PA. (3) The bacteria isolated from wounds of A and B groups was identified as PA. The subeschar bacteria count within PID 14 in A group, in which different amount of PA was inoculated, was mostly less than 1 × 105 CFU/g of tissue, while that in B group in which 1 × 109 CFU PA was inoculated was more than 1 × 105 CFU/g of tissue. (4) There was no obvious difference in wound healing time between wounds inoculated with different amount of PA and wounds treated with normal saline in A group ( with t value respectively 1.26, 0. 29, 1.07,P values all above 0.05 ). Wound healing time of wounds in B group, in which 1 × 109 CFU PA was inoculated, was longer as compared with that treated with normal saline [(22.5 + 1.0) d vs. ( 19.4 + 1.6) d, t =2.73, P ＜0. 05]. Conclusions In rat, deep partial-thickness burn wound inoculated with 1 × 109 CFU PA ATCC 27853 is a reliable model with high reproducibility for the study of infection of burn wound.     

雌激素对大鼠创面愈合过程中微血管形成的影响及其机制

目的 观察雌激素对大鼠全层皮肤缺损创面愈合过程中微小血管再形成的影响.方法 将40只雌性Wistar大鼠随机分成3组:单纯损伤组15只(A组),去卵巢组15只(B组)和假手术组10只(C组).8周后,在背部制成全层全损皮肤创伤愈合模型,分别于伤后1、3、7、14和21 d记录3组各个时间点伤腔容积,并在创缘取材.常规组织学观察皮肤愈合过程和微血管的数量,同时采用雌激素β受体(ER-β)进行免疫组织化学染色,观察微血管及周围细胞受体的染色.结果　比较各个时间点的伤腔容积,单纯损伤组和假手术组大鼠创面内肉芽组织形成量明显大于去卵巢组,组织学评分略高于去卵巢大鼠;而单纯损伤组和假手术组之间差异无统计学意义(P＞0.05).卵巢切除大鼠伤后各时间点,真皮内微小血管的数量较单纯损伤组减少,各组均于伤后7 d,随着成纤维细胞和毛细血管数目的 增加,肉芽组织生长达到高峰.同时,各损伤组创基内微小血管周围均出现较为幼稚的细胞,其表面可以表达ER-β,但卵巢切除组(159.91±12.65)稍弱于单纯损伤组(189.36±27.32),差异无统计学意义(P＞0.05).结论　大鼠去卵巢后创面愈合速度明显减慢与微血管形成有关,雌激素及其受体在该过程中有重要作用.

Abstract：

Objective To investigate the effect of estrogen on the revascularization of full-thickness skin wounds healing in rats. Methods Forty 2-month-old female Wistar rats were divided into three groups: control group (group A, pure injury,n = 15), ovariectomized group (group B,n = 15) and sham operation group (group C,n = 10 ). After 8 weeks, full-thickness wounds model, 18 mm in diameter, were made on the back of the each animal. The volume of wound was measured at each time point. The specimens of wound edge were obtained at the 1st, 3rd, 7th, 14th and 21st day after injury. The process of wound healing and the number of microvessels in the granulation tissue were evaluated by Haematine-Eosin (HE) staining. The immunohistochemical staining was used to detect the expression of estrogen receptor β (ER-β) in microvascular endothelial cells and their surrounding cells. Results To compare the volume of wound at each time point, the granulation tissue formation in group A and group C was significantly higher than that in group B, but there was no significant difference between group A and group C. The number of capillary blood vessels in group B was significantly reduced as compared with that in group A. The expression of ER-β was positive on the cells surface in all groups. ER-β expression in group B was weaker than in group A ( 159. 91 ± 12. 65 v. s 189. 36 ±27. 32,P ＜0. 05). Conclusion The delay of wound healing in ovariectomized rats has a great relationship with angiogenesis. Estrogen and its receptor might play important roles in the process.     

荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

目的 观察荷负电气溶胶治疗对大鼠烫伤创面愈合过程中白细胞介素(IL)-8、IL-10表达的影响,探讨荷负电气溶胶治疗促进创面愈合的作用机制.方法 制作SD大鼠深Ⅱ度烫伤模型,采用分组对照方法,将40只鼠随机分为治疗组(n1=20)和对照组(n2=20).治疗组应用荷负电气溶胶治疗,每次1.5 h,每天2次,直至创面愈合;对照组不作荷负电气溶胶治疗.伤后第1～11天分别取创面标本制作切片,采用免疫组织化学和图像分析方法,检测创面愈合过程中IL-8和IL-10表达水平.结果 创面平均愈合时间治疗组为(7.00±1.15)d,对照组为(9.00±1.34)d,治疗组创面愈合时间明显提前(P＜0.01).免疫组织化学显示,两组IL-8均在伤后第1天开始表达.主要位于多核粒细胞和单核细胞;第3天表达明显增多达高峰,并见大量成纤维细胞表达,治疗组的峰值明显低于对照组,差异有统计学意义(P＜0.01),第5～11天表达水平迅速下降.两组IL-10伤后第1天在淋巴细胞和单核细胞均有表达;第3天开始有角质细胞表达并达高峰,第5～11天表达水平缓慢下降,但治疗组要明显高于对照组,第3～11天差异有统计学意义(P＜0.01).结论 荷负电气溶胶治疗能有效抑制创面IL-8的表达及促进IL-10的表达,缩短炎症进程,从而加速创面愈合.

Abstract：

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

不同敷料组合对烧伤患者供皮区创面微环境的影响

Objective To investigate the effect of dressing materials in various combinations on bum wound microenvironment and healing condition. Methods Two hundred donor sites with wounds of 0.3 mm in depth in 186 burn patients, who needed skingrafting and admitted to our ward were enrolled in study, and they were divided into A ( with dressing composed of aiginate + cotton pad for donor area), B ( with dressing composed of vaseline gauze + cotton pad for donor area) , C (with dressing composed of algi-nate+ foam dressing for donor area), D ( with dressing composed of vaseline gauze + foam dressing for donor area) groups according to random table method. Effect of dressings on wound evaporation and pH value were observed. Bacterial colonization, degree of pain complained by patients after dressing change, and wound healing time in each group were compared. Results One hundred and eighty-four patients complied with the study, while 2 patients were excluded due to untimely falling-off of the dressing. Wound evaporaton in respectively, among them B group showed optimal effect of keeping humidity (P<0.01). Wound pH value in A, B, C, D groups was 7.22±0.06, 7.41±0.03, 7.05±0.03, 7.34±0.06, respectively, among them it was highest in B group. The positive rate of bacteria in D group was highest (22.4%), and lowest in C group (4.0%). Pain was lightest in C group (score was 0.98±0.12), and most serious in B group ( score was 8.14±0.82). The shortest wound healing time was seen in C group (6.7±0.8 d) , and longest in D group (15.6±3.5 d). Conclusions Application of various dressings on similar wounds can pro-duce different wound microenvironment, which is closely related to wound healing time. Compared with pH value, humidity is the more important factor for wound healing.     

不同敷料组合对烧伤患者供皮区创面微环境的影响

Objective To investigate the effect of dressing materials in various combinations on bum wound microenvironment and healing condition. Methods Two hundred donor sites with wounds of 0.3 mm in depth in 186 burn patients, who needed skingrafting and admitted to our ward were enrolled in study, and they were divided into A ( with dressing composed of aiginate + cotton pad for donor area), B ( with dressing composed of vaseline gauze + cotton pad for donor area) , C (with dressing composed of algi-nate+ foam dressing for donor area), D ( with dressing composed of vaseline gauze + foam dressing for donor area) groups according to random table method. Effect of dressings on wound evaporation and pH value were observed. Bacterial colonization, degree of pain complained by patients after dressing change, and wound healing time in each group were compared. Results One hundred and eighty-four patients complied with the study, while 2 patients were excluded due to untimely falling-off of the dressing. Wound evaporaton in respectively, among them B group showed optimal effect of keeping humidity (P<0.01). Wound pH value in A, B, C, D groups was 7.22±0.06, 7.41±0.03, 7.05±0.03, 7.34±0.06, respectively, among them it was highest in B group. The positive rate of bacteria in D group was highest (22.4%), and lowest in C group (4.0%). Pain was lightest in C group (score was 0.98±0.12), and most serious in B group ( score was 8.14±0.82). The shortest wound healing time was seen in C group (6.7±0.8 d) , and longest in D group (15.6±3.5 d). Conclusions Application of various dressings on similar wounds can pro-duce different wound microenvironment, which is closely related to wound healing time. Compared with pH value, humidity is the more important factor for wound healing.     

不同敷料组合对烧伤患者供皮区创面微环境的影响

Objective To investigate the effect of dressing materials in various combinations on bum wound microenvironment and healing condition. Methods Two hundred donor sites with wounds of 0.3 mm in depth in 186 burn patients, who needed skingrafting and admitted to our ward were enrolled in study, and they were divided into A ( with dressing composed of aiginate + cotton pad for donor area), B ( with dressing composed of vaseline gauze + cotton pad for donor area) , C (with dressing composed of algi-nate+ foam dressing for donor area), D ( with dressing composed of vaseline gauze + foam dressing for donor area) groups according to random table method. Effect of dressings on wound evaporation and pH value were observed. Bacterial colonization, degree of pain complained by patients after dressing change, and wound healing time in each group were compared. Results One hundred and eighty-four patients complied with the study, while 2 patients were excluded due to untimely falling-off of the dressing. Wound evaporaton in respectively, among them B group showed optimal effect of keeping humidity (P<0.01). Wound pH value in A, B, C, D groups was 7.22±0.06, 7.41±0.03, 7.05±0.03, 7.34±0.06, respectively, among them it was highest in B group. The positive rate of bacteria in D group was highest (22.4%), and lowest in C group (4.0%). Pain was lightest in C group (score was 0.98±0.12), and most serious in B group ( score was 8.14±0.82). The shortest wound healing time was seen in C group (6.7±0.8 d) , and longest in D group (15.6±3.5 d). Conclusions Application of various dressings on similar wounds can pro-duce different wound microenvironment, which is closely related to wound healing time. Compared with pH value, humidity is the more important factor for wound healing.     

不同敷料组合对烧伤患者供皮区创面微环境的影响

Objective To investigate the effect of dressing materials in various combinations on bum wound microenvironment and healing condition. Methods Two hundred donor sites with wounds of 0.3 mm in depth in 186 burn patients, who needed skingrafting and admitted to our ward were enrolled in study, and they were divided into A ( with dressing composed of aiginate + cotton pad for donor area), B ( with dressing composed of vaseline gauze + cotton pad for donor area) , C (with dressing composed of algi-nate+ foam dressing for donor area), D ( with dressing composed of vaseline gauze + foam dressing for donor area) groups according to random table method. Effect of dressings on wound evaporation and pH value were observed. Bacterial colonization, degree of pain complained by patients after dressing change, and wound healing time in each group were compared. Results One hundred and eighty-four patients complied with the study, while 2 patients were excluded due to untimely falling-off of the dressing. Wound evaporaton in respectively, among them B group showed optimal effect of keeping humidity (P<0.01). Wound pH value in A, B, C, D groups was 7.22±0.06, 7.41±0.03, 7.05±0.03, 7.34±0.06, respectively, among them it was highest in B group. The positive rate of bacteria in D group was highest (22.4%), and lowest in C group (4.0%). Pain was lightest in C group (score was 0.98±0.12), and most serious in B group ( score was 8.14±0.82). The shortest wound healing time was seen in C group (6.7±0.8 d) , and longest in D group (15.6±3.5 d). Conclusions Application of various dressings on similar wounds can pro-duce different wound microenvironment, which is closely related to wound healing time. Compared with pH value, humidity is the more important factor for wound healing.     

不同敷料组合对烧伤患者供皮区创面微环境的影响

Objective To investigate the effect of dressing materials in various combinations on bum wound microenvironment and healing condition. Methods Two hundred donor sites with wounds of 0.3 mm in depth in 186 burn patients, who needed skingrafting and admitted to our ward were enrolled in study, and they were divided into A ( with dressing composed of aiginate + cotton pad for donor area), B ( with dressing composed of vaseline gauze + cotton pad for donor area) , C (with dressing composed of algi-nate+ foam dressing for donor area), D ( with dressing composed of vaseline gauze + foam dressing for donor area) groups according to random table method. Effect of dressings on wound evaporation and pH value were observed. Bacterial colonization, degree of pain complained by patients after dressing change, and wound healing time in each group were compared. Results One hundred and eighty-four patients complied with the study, while 2 patients were excluded due to untimely falling-off of the dressing. Wound evaporaton in respectively, among them B group showed optimal effect of keeping humidity (P<0.01). Wound pH value in A, B, C, D groups was 7.22±0.06, 7.41±0.03, 7.05±0.03, 7.34±0.06, respectively, among them it was highest in B group. The positive rate of bacteria in D group was highest (22.4%), and lowest in C group (4.0%). Pain was lightest in C group (score was 0.98±0.12), and most serious in B group ( score was 8.14±0.82). The shortest wound healing time was seen in C group (6.7±0.8 d) , and longest in D group (15.6±3.5 d). Conclusions Application of various dressings on similar wounds can pro-duce different wound microenvironment, which is closely related to wound healing time. Compared with pH value, humidity is the more important factor for wound healing.     

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.     

An experimental study on the repair of full skin loss of nude mice with composite graft of epidermal stem cells

This study is to constitute a composite skin substitute with epidermal stem cells (ESCs) and fibroblasts on collagen sponge. ESCs were selected by rapid attachment to collagen IV for 10 min. Collagen was extracted from rat's tail. The matrix lattice was fabricated by freeze-dryer and cross-linked with glutaraldehyde. Fibroblasts were inoculated on collagen sponge and cultured for 1 week prior to inoculation of ESCs. Having cultured for 2 weeks in submerged culture, the bioengineered tissue was raised to the air-liquid interface and cultured for 2 weeks. The artificial skin was then grafted onto full skin loss wounds of nude mice. Collagen sponge membrane lacking cell inoculation and an artificial skin with epidermal cells (ECs) and fibroblasts were used as controls. The wounds were observed daily. Tissue samples were harvested and examined by means of histology, immunohistochemistry and electron microscopy. The wounds in the test group healed at a significantly faster rate than controls, with good skin appearance and minimal scar formation. The control group showed delayed wound healing and intensive wound contraction as compared to the test group. Thus the skin substitute with ESCs seemed to be a good equivalent.     

不同剂量rhGM-CSF凝胶促进大鼠深Ⅱ度烧伤创面愈合的实验研究

目的：观察不同剂量重组人粒细胞-巨噬细胞集落刺激因子（rhGM-CSF）凝胶（商品名为金扶宁）对SD大鼠深Ⅱ度烫伤创面愈合的促进作用并筛选最佳剂量.方法：42只SD大鼠,雌雄各半,随机分为7组：模型组,凝胶基质对照组,金扶宁100、250、500、750和1 000 mg/cm^2治疗组.每只大鼠背部制备两个深Ⅱ度烫伤创面,凝胶基质对照组创面于伤后1～10 d每日涂抹凝胶基质100 mg/cm^2,然后于伤后12、14、17和19 d间断创面涂抹凝胶基质100 mg/cm^2;不同剂量金扶宁治疗组创面涂沫相应剂量金扶宁,给药方法及时间同凝胶基质对照组;模型组不予处理.伤后3、7、10、14、17、21 d分别测量痂皮形成及脱落创面数,计算创面上皮化率、创面愈合率,伤后21 d行组织学检查.结果：与模型组和凝胶基质对照组相比,金扶宁各组对烫伤后7～10 d前后创面的融痂脱痂效果非常明显,以250～500 mg/cm^2治疗组疗效最佳（P〈0.05）.治疗后7～10 d,金扶宁各组新生上皮较模型组和凝胶基质对照组均显著增多（P〈0.05）;治疗后14 d,金扶宁500 mg/cm^2治疗组上皮化率[（55.46±6.13）%]较模型组[（48.81±4.99）%]和凝胶基质对照组[（43.17±5.65）%]仍然显著增多,并优于其他剂量治疗组（P〈0.05）.治疗后10～14 d,金扶宁250～500 mg/cm2治疗组创面愈合率较模型组、凝胶基质对照组和其他剂量治疗组显著增高（P〈0.05）;治疗后21 d,金扶宁250～1 000 mg/cm^2治疗组毛囊再生数量较模型组[（1.531±0.374）/mm2]和凝胶基质对照组[（1.423±0.346）/mm^2]显著增高,以500～750 mg/cm^2治疗组[分别为（7.145±1.153）/mm2和（6.250±0.768）/mm2]明显（P〈0.05）.结论：rhGM-CSF凝胶可以促进痂皮形成和脱落、加快上皮生长、促进毛囊再生,增加烧伤创面修复速度并改善修复质量,以500 mg/cm^2剂量治疗效果最佳.     

Topical embryonic stem cells enhance wound healing in diabetic rats

The effects of embryonic stem cells (ESCs) on diabetic wound healing were investigated using an excisional skin wound model in 110 diabetes‐induced rats. We transplanted a clonal population of ESCs (5 × 10⁶) by topical injection into full thickness skin wounds. Four study groups were used; nondiabetic rats as a control, non‐insulin controlled diabetic rats not treated with ESCs, insulin controlled diabetic rats not treated with ESCs, and insulin controlled diabetic rats treated with ESCs. Five rats in each experimental group were sacrificed on days 1, 5, 10, 15, and 20 after wounding. Wounds images were acquired daily and wound sizes were calculated. We measured the mRNA levels of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF), and fibronectin levels in extracellular matrix, and assessed wound healing by assessing histological parameters of epidermal regeneration, granulation tissue thickness, and angiogenesis. In the ESC‐treated group, wound sizes were significantly smaller than in the insulin controlled diabetic group not treated with ESCs on days 5 and 10 (p < 0.05), and EGF and VEGF levels were markedly higher on days 5 and 10, fibronectin levels on day 5 after injection. All histological scores in the ESC‐treated group were significantly higher than those of the insulin controlled diabetic group on day 5 (p < 0.05). Our results shows that topical ESCs enhance diabetic wound healing during the early stage, and suggest that ESCs transplantation offers a novel therapeutic modality for the treatment of diabetic wounds. © 2011 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 29: 1554–1562, 2011     

静电纺丝血管外支架抑制静脉移植物内膜增生

目的 探讨聚-3-羟基丁酸与聚-4-羟基丁酸聚酯[P(3HB-co-4HB)]静电纺丝血管外支架预防静脉移植物内膜增生的机制.方法 静电纺丝技术制备P(3HB-co-4HB)血管外支架.SD大鼠随机分成2组,每组24只,建立颈外静脉-颈总动脉血管移植模型:无支架(Ns)组和非限制支架(PS)组.分别于术后3、7、14、28 d取静脉移植物标本,计算机图像系统分析内膜、中膜厚度和面积,免疫组织化学检测静脉移植物增殖细胞核抗原(PCNA)表达.逆转录-聚合酶链反应(RT.PCR)检测静脉移植物血小板源性生长因子.BB(PDGF-BB)和组织因子(TF)mRNA表达.结果 术后7、14、28 d支架组静脉移植物内膜与中膜的厚度和面积明显低于无支架组(P<0.05).两组PCNA表达均增加并在术后14 d最高,而支架组在术后7、14 d PCNA表达低于无支架组(P<0.05).术后3、7d支架组PDGF.BB和TFmRNA表达明显低于无支架组(P<0.01).结论 静电纺丝P(3HB-co-4HB)血管外非限制性支架能抑制静脉移植物内膜增生,并通过降低静脉移植物管壁早期组织因子表达发挥作用.     

铜绿假单胞菌感染大鼠皮肤创面愈合过程中TGF-β1及胶原蛋白含量的变化

目的 探讨大鼠慢性皮肤溃疡创面感染铜绿假单胞菌后,TGF-β1和胶原Ⅰ、Ⅲ蛋白表达的变化.方法 24只8周龄雌性Wister大鼠随机分为单纯创面组(A组)和创面+铜绿假单胞菌接种组(B组).分别在术后第1、3、7、10天观察创面上皮化率、收缩率及中性粒细胞情况;并采用ELISA方法测定创面第1、3、7、10天TGF-β1和胶原Ⅰ、Ⅲ蛋白的表达情况.结果 A组上皮化率在第7天高于B组,收缩率低于B组.随着时间的延长,A组中性粒细胞在第3天增加到最多,随后逐渐减少,而B组中性粒细胞第1天达到最多.2组TGF-β1表达在术后呈上升趋势,B组TGF-β1在第3天降低,随着时间延长回升,第7天2组比较差异有统计学意义(P＜0.05).胶原Ⅰ、Ⅲ蛋白表达随着时间的延长呈下降趋势,B组胶原Ⅲ蛋白表达在第7、10天差异有统计学意义(P＜0.05).A组第1、3天胶原Ⅰ、Ⅲ蛋白表达高于B组,在第7、10天则低于B组,且胶原Ⅲ蛋白在第3天显示A组高于B组,差异有统计学意义(P＜0.05).结论 皮肤溃疡创面感染铜绿假单胞菌后延迟了TGF-β1和胶原Ⅰ、Ⅲ蛋白的表达,可能影响创面的正常愈合.

Abstract：

Objective To explore the different expression of TGF-β1 and collagen during the healing process of wound infected by pseudomonas aeruginosa(PAO1).Methods 24 female Wister rats were randomly divided into pure wound group(group A)and wound+PAO1 group(group B).The reepithelial rate,shrinkage rate and neutrophils number on the wounds were observed on the 1st,3rd,7th and 10th day after operation.The expression of TGF-β1 and collage Ⅰ,Ⅲ was also detected.Results On the 7th day,the re-epithelial rate in group A was higher than that in group B,while the shrinkage rate in group A was lower than that in group B.The neutrophils number increased to peak on the 1 st day in group B,but on the 3rd day in group A.The TGF-β1 expression increased after operation in both groups,but it decreased in group B on the 3rd day and re-increased after that.The TGF-β1 expression was significantly different between the two groups on the 7th day(P＜0.05).The expression of collagen Ⅰ and Ⅲ decreased during healing.The expression of collagen Ⅲ in group A was higher on the 3rd day and was lower on the 7th and 10th day than that in group B,showing a significant difference(P＜0.05).Conclusions PAO1 infection could delay the expression of TGF-β1 and collagen Ⅰ,Ⅲon wound,which may interfere the healing process of wound.     

富血小板纤维蛋白修复大鼠骨外露创面的实验研究

目的探讨富血小板纤维蛋白(PRF)治疗大鼠骨外露创面的疗效。方法取10~12周龄,250~300 g的雄性SD大鼠24只,应用随机数字表法将其分为3组,每组8只。将所有SD大鼠前头盖骨区域1.5 cm×1.5 cm皮肤全层切除及并去除颅骨表面的骨膜。PRF组创面覆盖PRF;脂肪移植组创面覆盖等量脂肪颗粒;对照组创面覆盖等量生理盐水。分别于第4、7和11天对创面换药。使用Image J软件对骨外露面积进行量化评估。应用HE染色和马松三色染色评估创面新生血管和胶原沉积情况。酶联免疫吸附实验检测创面肉芽组织生长因子水平。组间比较采用随机资料单因素方差分析。结果术后第4天时,PRF组大鼠骨外露比例为57.99%±11.29%;脂肪移植组骨外露比例为45.92%±9.55%;对照组骨外露比例为77.73%±5.57%。第7天时,PRF组大鼠骨外露比例为4.29%±2.28%;脂肪移植组骨外露比例为29.52%±6.33%;对照组骨外露比例为36.90%±8.43%。第11天时,仅PRF组的骨外露创面完全被肉芽组织覆盖,脂肪移植组及对照组均存在不同程度的骨外露,分别为10.15%±1.49%和21.69%±2.40%。PRF组在各时间点创面骨外露面积均显著低于对照组,差异具有统计学意义(P<0.01)。在第7天和第11天时,PRF组的骨外露比例显著低于脂肪移植组,但是在第4天时,脂肪移植组的骨外露比例却低于PRF组,差异均具有统计学意义(P<0.001)。第11天时,HE染色结果显示PRF组的新生微血管密度为(10.37%±0.49%)显著高于脂肪移植组(4.86%±0.83%)和对照组(2.91%±0.31%)(P<0.05),马松三色染色结果显示PRF组的胶原纤维最丰富。酶联免疫吸附实验结果提示第11天时PRF组的生长因子水平均明显升高,与脂肪移植组和对照组比较,差异均具有统计学意义(P<0.05)。结论PRF作为一种取材方便的治疗手段,对修复创面具有良好的疗效,可加快骨外露创面愈合。