Objective To explore the expression characteristics of chaperone interacting protein (CHIP) in normal, scar and chronic ulcer tissues and its relationship with wound healing. Methods Twenty biopsies including scar tissues(n=8), chronic ulcer tissues(n=4) and normal tissues(n=8)were used in this study. The immunohistochemical staining (power visionTMtwo-step histostaining reagent) was used to explore the amount and expression characteristics of such protein.Results The positive expression of CHIP was observed in fibroblasts, endothelial cells and epidermal cells in dermis and epidermis. It was not seen ininflammatory cells. The expression amount of CHIP in scar tissues, chronic ulcer tissues and normal tissues was 89%, 83% and 17% respectively. Conclusion Although the function of CHIP is not fully understood at present, the fact that this protein is expressed only at the mitogenic cells indicates that it may be involved in mitogenic regulation during wound healing.
Abstract In order to understand the expression change of epidermal growth factor (EGF) gene and its distribution in tissue duringwound healing, 12 Winstar rats were divided into 4 groups. In each side of the back of every rat, 4 wounds of 1.5cm×1.5cm in size were made. After 4,8,12,16 days, one group of rats was sacrificed and tissues from wound were collected. By DIG-labelled probe hybridization in situ technique, EGF gene mRNA was detected. It was shown that EGF gene expression was evident during the whole stage of woundhealing, and the peak was in 8th day. It suggested that the promotion of EGF gene expression may lead to wound healing earlier.
Objective To compare and research the process of woundhealing in occlusive moist environment and dry environment on the skin donor site. Methods The wound healing of adult skin donor site was studied by clinical observation, histological and electromicroscopical examinations on the operative day and the 1st, 2nd, 3rd, 4th, 7th days postoperatively, each skin donor site was divided into two parts: occlusive environment and dry environment. Results The wounds of occlusive moist environment healed faster than those of dry environment; thefibroblasts were more active and activated earlier, revascularization and re-epithelialization happened earlier and more quickly. Conclusion In occlusive environment, more active fibroblasts can accelerate granulation growth; quicker regenerative capillaries bring more nourishment; quicker re-epithelialization accelerates the wound healing.
【Abstract】 Objective To investigate the effects on forming of hypertrophic scar after BMSCs infected with adenovirus carrying TGF-β3c2s2 were transplanted into the wound of animal scar model. Methods The third passage of rabbit’ s BMSCs were infected with 150 mutiple infection, and were cultured 24 hours. The concentration of the BMSCs infected with recombinant adenovirus containing the TGF-β3c2s2 gene was 1×105cell/mL. The purified and evaporated recombinant adenovirus grains containing the TGF-β3c2s2 gene were diluted by DMEM/F12 (without FBS) to 1×108 pfu/mL. The animal scar model of the standard Japanese big ear rabbit was establ ished. Eighty wounds were generated on the gastroside of ear and were randomized to 4 groups in each rabbit, which were divided into 3 control groups (A: control, B: Ad-TGF-β3c2s2, C: BMSCs) and 1 experimental group (D: BMSCs/Ad-TGF-β3c2s2). Then the wounds were tranplanted with cells. On 45 days and 90 days after wounded, thicknessand hardness of scars were measured with color ultrasound diagnostic unit and especial measurement for skin and scar hardness. On 21, 45 and 90 days, three specimens were harvested respectively for further histological study. Results The wound of groups A, B, C gradually formed the different degree scars after epithel ial ization. The hyperplasty of scars reached peak on 45 days after wounded and lasted about 90 days. There was no prominent scar formed in group D during the whole observed procedure. Thickness and hardness of scar of group D and group E were approximate on 45 days and 90 days. Thickness and hardness of scar of groups A, B and C were lower than those of group D (P lt; 0.01), and group B showed more lower than group A and group C (P lt; 0.01). Disorder structure and overlapping arrangement, enlargement collagen fibers were showed in the HE histological sections of the scars of groups A, C. The structure of the scars of groups B, C were similar to Group E. The constitutionsof groups A, B, C, D on 90 days resembled to each one on 45 days. In section of immunohistochemistry after wounded on21 days and 45 days, positive expressions of BrdU in nucleus of Groups C, D were observed. Negative expressions of BrdU in Groups A, B, E were showed. Conclusion BMSCs with Ad-TGF-β3c2s2 gene transplanted into wound could inhibit the forming of hypertrophic scar.
Objective Human acellular amniotic membrane (HAAM) contains collagens, glucoproteins, proteinpolysaccharide,integrin, and lamellar, which can supply rich nutrition to cell prol iferation and differentiation. To explore the possibil ity of HAAM with adi pose-derived stem cells (ADSCs) as a good engineered skin substitute for repairing skin defect. Methods Primary ADSCs were obtained from inguinal fat of 30 healthy 4-month-old SD rats, male or female, weighing 250-300 g, and cultured in vitro and purified. The 3rd passage ADSCs were used to detect CD44, CD49d and CD34 by immunocytochemistry staining. After physical and trypsin preparation, the HAAM was observed by HE staining and scanning electron microscope(SEM) respectively. ADSCs were seeded on epithel ial side of HAAM at the density of 2 × 105/cm2, cocultured, and observed by SEM at different time. MTT test was used to detect viabil ity of cells that seeded on HAAM, the group without HAAM was used as control. Thirty SD rats were made models of full-thickness skin wound and randomly divided into three groups (A, B, and C). Wound was repaired with HAAM/ADSCs composites in group A, with HAAM in group B, and with gauze as control in group C. The rats underwent postoperative assessment of wound heal ing rate and histological observation at the 1st, 2nd, and 4th weeks. Results HE staining showed that the 3rd passage ADSCs was spindle-shaped with an ovoid nucleus which located in the middle of cell; the immunocytochemistry staining showed positive result for CD44 and CD49d and negative result for CD34. There were no residues of cells in the HAAM by HE staining. SEM showed that there were different structures at the two sides of HAAM；one side had compact reticular structure and the other side had fibrous structure. After 3 days of co-culture, ADSCs showed good growth on HAAM; the cells were closely packed onto the HAAM, attached firmly and prol iferated to confluence on the stromal surface of HAAM. MTT test showed that the cells on the HAAM grew well and had b prol iferation vital ity. There was no significant difference between ADSCs cultured in the HAAM and control group (P gt; 0.05). One, 2, 4 weeks after graft, there were significant differences in wound heal ing rate between group A and groups B, C (P lt; 0.05), between group B and group C (P lt; 0.05). HE staining showed that wound healed faster in group A than in groups B, C. Cytokeratin 19 (CK19) immunohistochemical statining showed that there were more CK19 positive cells in group A than in groups B, C. Conclusion The graft of HAAM with ADSCs plays an effective role in promoting the repair of full-thickness skin wound
【摘要】 目的 探讨中成药参附注射液对大鼠深Ⅱ度烧伤创面愈合的影响。 方法 选用16只Sprague-Dawley（SD）大鼠，随机分为实验组、对照组，每组各8只，建立20%体表面积深Ⅱ度烫伤模型。烫伤后即刻及此后每天，实验组大鼠以参附注射液20 mL/kg腹腔注射给药，1次/d，连续给药5 d；对照组给予注射等剂量的生理盐水。分别于致伤后的7、14 d取创面组织块，光学显微镜观察成纤维细胞及胶原纤维生长情况，电子显微镜观察致伤后14 d成纤维细胞细胞器情况，分别计算烧伤后第7、14天两组的创面愈合率，并观察比较两组创面愈合的时间。 结果 所有选入实验的SD大鼠均存活至实验结束。实验组、对照组术后第7天创面愈合率分别为（36.34±2.55）%及（33.13±2.62）%，两组差异有统计学意义（t=2.486，P=0.027）；实验组、对照组术后第14天创面愈合率分别为（75.71±2.29）%及（72.36±2.85）%，两组差异有统计学意义（t=2.590，P=0.022）；实验组、对照组创面愈合时间分别为（20.88±1.36）、（22.94±2.16） d，两组差异有统计学意义（t=-2.286，P=0.0395）。伤后7、14 d创面组织块切片光学显微镜观察发现，实验组肉芽组织及胶原纤维较多，排列更有序，表皮基底细胞增殖活跃，炎症反应较轻；伤后14 d时电子显微镜观察发现，实验组成纤维细胞的细胞器更丰富，分泌胶原更多，实验组创面愈合情况优于对照组。 结论 腹腔注射参附注射液可以促进烧伤创面的愈合，其可能的机制为清除氧自由基，抗脂质过氧化。【Abstract】 Objective To explore the effect of Shenfu injection on promoting healing of deep partial-thickness burn wound in rats. Methods Sixteen Sprague-Dawley (SD) rats were randomly selected, and deep partial-thickness burn with 20% of the body surface was inflicted. The rats were randomly divided into experimental group and control group with eight in each group. Rats in the experimental group were treated with abdominal injection of Shenfu injection at a dose of 20 mL/(kg•d) for five days continually, and rats in the control group were treated with the same dose of 0.9% saline solution. The growth of granulation tissue and collagen fibers were evaluated under light microscope at the seventh and the fourteenth day. The growth of fibroblast was observed under transmission electron microscope at the fourteenth day. The cure rate of both groups of rats at the seventh and fourteenth day was calculated. Then we compared their healing time and the cure rate of the traumatic wound respectively. Results All rats had survived until wound healing. The cure rate at the seventh day for the experimental group and the control group was (36.34±2.55)% and (33.13±2.62)%, and their difference was statistical (t=2.486, P=0.027); At the fourteenth day, the cure rate was respectively (75.71±2.29)% and (72.36±2.85)% with a significant difference between each other (t=2.590, P=0.022). The healing time of the experimental group (20.88±1.36) was significantly shorter than that of the control group (22.9±2.16) (t=-2.286, P=0.040). At the seventh and fourteenth day, light microscope observation showed that the growth of granulation tissue and collagen fibers for rats in the experimental group were much more than that in the control group, the basale cell proliferation was more active, and inflammation was slighter. Through transmission electron microscope, we observed more fibroblast and collagen in the experimental group, which showed a better cure than the control group. Conclusion Shenfu injection can significantly promote wound healing of deep partial-thickness burn. It may possibly get this effect through anti-oxidation.
OBJECTIVE: With the recognization of the mechanism of wound healing, some topical agents are created and applied in trauma to improve the healing rate of wounds. The main purpose of this study is to investigate the effect of some topical agents on the healing rate of deep second-degree burn wounds. METHODS: One thousand five hundred and sixty-three patients with deep second-degree burn wounds(total burn surface area lt; or = 10%) were involved in this study from January 1982 to December 1999. According to the application time of different treating measures including supplement of Zn, application of growth factors and collagenase, the patients were divided into 3 groups, wound healing rates were compared. RESULTS: Before 1991, none of special topical agents were used, and the healing time of deep second-degree burn wounds was(23.8 +/- 3.5) days. From 1991 to 1996, with the topical application of SD-Ag-Zn, which can provide Zn for cells taking part in wound healing, the healing time of deep second-degree burn wounds was (20.6 +/- 3.2) days, earlier than no special topical agents (P lt; 0.05). From 1997 to 1999, growth factors such as basic fibroblast growth factor(bFGF) and epithelial growth factor (EGF) and collagenases were applied in wound treatment combining with SD-Ag-Zn, wound healing time was (16.2 +/- 2.8) days, earlier than no special topical agents (P lt; 0.01) and simple SD-Ag-Zn application (P lt; 0.05). CONCLUSION: It indicates that the improvement of topical agents can accelerate wound healing speed.
ObjectiveTo investigate the mechanism of impeded wound healing by exogenous hyaluronan (HA). Methods Wound healing models were established on 18 adult rabbit ears, which were randomly divided into 3 groups, the 2% HA treated-group (group A), the 1% HA treated-group(group B), and the PBS control-group (group C). The process of wound healing was observed morphologically and histologically. The expression of α-smooth muscle actin in fibroblast was measured by immunohistochemical method. Results ①The mean values of wound healing time of groups A, B and C were (11.7±0.6), (11.3±0.6), and (10.8±1.0) days respectively. Wound contraction was greater in group C than in group A and group B. ②Compared with PBS controls, the collagen fibril was slender and arrayed regularly in HA treated wound. ③ The expression of α-smooth muscle actin was greater in group C than in groups A and B. Conclusion It is one of reasons of impeded wound healing that exogenous HA inhibits the expression of α-smooth muscle protein and wound contraction. There exists dose-dependant effect.
OBJECTIVE: To explore an optimal method of recombinant human epidermal growth factor(rhEGF) application on the burn wounds of superficial II degree and profound II degree for accelerating its healing. METHODS: There were 180 burn wounds in 60 patients with the self-corresponding wound of the same degree as controls. The wounds of all patients were divided three regions(A, B, C). The wounds were treated once a day with 1% SD-Ag in region A as controls, with rhEGF(40 U/cm2) in region B, and with a combination of rhEGF(40 U/cm2) and Su Yu Ping (5 g) in region C. The wound healing time was recorded and compared. RESULTS: In regions A, B and C, the healing time of superficial II degree wound was (13.20 +/- 2.40) days, (10.20 +/- 2.20) days and (8.72 +/- 2.31) days (P lt; 0.01); that of profound II degree wound was (20.10 +/- 3.40) days, (17.20 +/- 3.12) days and (15.10 +/- 3.81) days respectively (P lt; 0.01, P lt; 0.05). The healed wound of profound II degree was elastic and tough in regions B and C, while that was not elastic and tough, and congestive in region A. CONCLUSION: The above results indicate that rhEGF can enhance burn wound healing markedly and that a combination rhEGF and Su Yu Ping has more significant effect than rhEGF alone and is recommended for clinical application.