重组人生长激素促进烧伤病人创面愈合机制初探

目的观察严重烧伤病人接受重组人生长激素（ｒｈＧＨ）治疗后血清生长激素（ＧＨ）、胰岛素样生长因子－１（ＩＧＦ－１）、创面ＤＮＡ合成。细胞增殖活性及表皮生长因子受体（ＥＧＦＲ）表达的变化，探讨生长激素促进烧伤创面愈合的机制。方法３３例严重烧伤病人随机分成ｒｈＧＨ组（１８例）和对照组（１５例）。ｒｈＧＨ组每晚１０点接受ｒｈＧＨ皮下注射（０．１ｍｇ· ｋｇ~(－１)），疗程 １４ｄ。ＥＬＩＳＡ检测血清 ＧＨ和 ＩＧＦ－１浓度；流式细胞计检测细胞 ＤＮＡ增殖指数（ＰＩ）和合成期细胞百分数（ＳＰＦ）；免疫组化学检测增殖细胞核抗原（ＰＣＮＡ）。ＥＧＦＲ、细胞角蛋白（ＣＫ）的表达。结果ｒｈＧＨ组血清ＧＨ、ＩＧＦ－１浓度较高，创面ＰＩ、ＳＰＦ明显高于用药前和对照组。免疫组化显示ＰＣＮＡ、ＥＧＦＲ、ＣＫ的表达明显增加。结论ｒｈＧＨ可以提高严重烧伤病人血清ＧＨ、ＩＧＦ－１水平，促进烧伤创面细胞ＤＮＡ合成和上皮细胞增殖，增加 ＥＧＦＲ表达，加速创面上皮化，促进烧伤创面愈合。     

大鼠浅度烫伤创面愈合与相关生长因子及受体的基因表达

目的探讨浅Ⅱ度烧伤创面愈合过程中相关生长因子及其受体的作用。方法采用大鼠浅Ⅱ度烫伤模型,观察了大鼠浅Ⅱ度烫伤后创面愈合过程中组织学、炎性细胞、表皮细胞周期的变化,检测了创面相关生长因子及受体的基因表达。结果炎性细胞到达创面依次为中性粒细胞、巨噬细胞和淋巴细胞;表皮细胞周期变化为伤后３天细胞增殖活跃,伤后７天细胞分裂明显增多;伤后１,３天ＰＤＧＦ（血小板源性生长因子）、ＰＤＧＦＲ（血小板源性生长因子受体）和ＥＧＦＲ（表皮生长因子受体）基因表达明显增强,伤后３,５天ＥＧＦ（表皮生长因子）、ＴＧＦβＲ２（转化生长因子β受体α）基因表达增强,伤后５,７天ＴＧＦβＲ１（转化生长因子β受体１）基因表达增强,伤后７,１０天ＴＧＦβ１（转化生长因子β）基因表达增强。结论烧伤诱导相关生长因子及受体的基因表达,并且具有时相性和可逆性;相关生长因子基因表达与表皮细胞周期间可能存在相互调控作用,生长因子是调控创面愈合的主要因素。     

表皮生长因子促进皮肤创面愈合的研究

为了观察外源性表皮生长因子对创面愈合的影响，在３０例大鼠皮肤创面愈合模型中，局部外用表皮生长因子（ＥＧＦ），以软膏基质作为对照，分别于１，２周测量创面面积，测定创面组织中的ＤＮＡ、蛋白质及羟脯氨酸含量，记录创面完全愈合时间。结果表明，实验组与对照组创面完全愈合的时间分别为（１４．６±１．２）天和（１８．５±２．０６）天（Ｐ＜０．０１），ＥＧＦ能显著增加组织中的ＤＮＡ、蛋白质及羟脯氨酸含量（Ｐ＜０．０１）。结果提示，ＥＧＦ能显著加速创面的修复，缩短愈合时间。     

烧伤创面应用生长因子后的DNA分析

将基因重组的人表皮细胞生长因子（ｒｈ－ＥＧＦ）、酸性成纤维细胞生长因子（ｒ－ａＦＧＦ）、碱性成纤维细胞生长因子（ｒ－ｂＦＧＦ）和牛脑提取液（ｂＢＥ）外用于烧伤创面，旨在探讨皮肤组织和细胞ＤＮＡ的含量及周期变化。实验结果显示：烧伤创面经应用ｒｈ－ＥＧＦ，ｒ－ａＦＧＦ，ｒ－ｂＦＧＦ和ｂＢＥ后第５天与生理盐水组（ＮＳ）相比，创面愈合率、表皮细胞和全厚层皮肤组织的ＤＮＡ含量均高于ＮＳ组（Ｐ＜０．０１）；全厚层皮肤ＤＮＡ的流式细胞仪分析表明，应用生长因子各组的Ｓ期细胞数百分比均高于ＮＳ组（Ｐ＜０．０１），细胞分裂增殖旺盛，Ｇ期细胞不断进入Ｓ期，使Ｓ期细胞数增加，ＤＮＡ的合成和含量增加。提示：烧伤创面外用细胞生长因子可促进表皮细胞的增殖，加速创面的愈合。     

PDGF,TGF—β1在人断层皮片供区创面愈合过程中的基因表 …

目的 探讨血小板源生长因子（ＰＤＧＦ）、转化生长因子－β１（ＴＧＦ－β１）对伤口愈合的影响。方法 用原位杂交技术观察人断层皮片供皮区创面愈合过程中内源性ＰＤＧＦ、ＴＧＦ－β１的基因表达。结果 发现人断层皮片供皮区创面愈合过程中内源性ＰＤＧＦ、ＴＧＦ－β１的基因有明显表达。结论 其表达规律性在创面愈合过程中可能起着重要的调控作用。     

神经生长因子调控烧伤创面的实验研究

目的 研究神经生长因子（ＮＧＦ）在烧伤创面愈合的作用,探索烧伤创面愈合机制。方法２０ｋｇ小白家猪６只为实验对象。用控温控压电烫仪在每只猪背部制成２４个直径为２．５ｃｍ的圆形深Ⅱ度烧伤创面。分为四组,分别为创面局部应用１、２．５和５μｇ／ｍｌ的ＮＧＦ实验组和不含ＮＧＦ的生理盐水对照组。在用药后３、５和９天分别取创面组织进行表皮生长因子（ＥＧＦ）受体,ＥＧＦ,ＮＧＦ受体,ＮＧＦ,ＣＤ６８,ＣＤ３免疫组     

PDGF、TGF-β_1在人断层皮片供区创面愈合过程中的基因表达及意义

目的探讨血小板源生长因子（ＰＤＧＦ）、转化生长因子－β１（ＴＧＦ－β１）对伤口愈合的影响。方法用原位杂交技术观察人断层皮片供皮区创面愈合过程中内源性ＰＤＧＦ、ＴＧＦ－β１的基因表达。结果发现人断层皮片供皮区创面愈合过程中内源性ＰＤＧＦ、ＴＧＦ－β１的基因有明显表达。结论其表达规律性在创面愈合过程中可能起着重要的调控作用     

创面愈合过程中内源性EGF变化的研究

为探讨创面愈合的机理，本实验采用免疫组织化学方法，对大鼠中厚皮片供皮区创面愈合过程中第４、８、１２和１６天创面内源性表皮生长因子（Ｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＥＧＦ）的变化进行了研究。结果表明，创面愈合过程中内源性ＥＧＦ含量表现有规律性变化，以伤后第８天含量最多，伤后早期和晚期次之，创面愈合后其含量进一步减少。结论：创面愈合过程中，内源性ＥＧＦ的变化促进了创面愈合，是创面愈合的机理之一，结合内源性ＥＧＦ变化，合理外用ＥＧＦ对创面愈合可能会取得促进的效果。     

白芨胶载HMW－NGF促进伤口愈合的实验研究

用中药白芨胶（ｂｌｅｔｉｌａｓｔｒｉａｔａｇｅｌａｔｉｎ，ＢＳＧ）作为外源性高分子神经生长因子（ｈｉｇｈｍｏｌｅｃｕｌａｒｗｅｉｇｈｔｎｅｒｖｅｇｒｏｗｔｈｆａｃｔｏｒ，ＨＭＷ－ＮＧＦ）的载体，观察促进大鼠伤口愈合的作用。实验分两部分进行：①将生理盐水、ＢＳＧ、ＨＭＷ－ＮＧＦ及ＨＭＷ－ＮＧＦ＋ＢＳＧ分别加入无血清培养基中，观察其对鸡胚背根神经节的影响；②选用ＳＤ大鼠４０只，分Ａ、Ｂ、Ｃ及Ｄ四组，在各大鼠背部切取２ｃｍ的伤口，一组大鼠伤口不用药，其余三组伤口分别给予外用ＢＳＧ、ＨＭＷ－ＮＧＦ及ＨＭＷ－ＮＧＦ＋ＢＳＧ，每天用药１次。在用药后第３及第１０天测量伤口面积，并用图象分析仪计算伤口面积，用光学显微镜观察细胞渗出类型和数量，并观察伤口愈合时间。结果表明：ＢＳＧ对ＨＭＷ－ＮＧＦ的活性无影响；ＢＳＧ、ＨＭＷ－ＮＧＦ及ＨＭＷ－ＮＧＦ＋ＢＳＧ都能促进伤口愈合和增加创面渗出多形核白细胞、单核细胞和成纤维细胞，尤以ＨＭＷ－ＮＧＦ＋ＢＳＧ的作用最为显著。可以得出以下结论：ＨＭＷ－ＮＧＦ＋ＢＳＧ能显著加速伤口愈合     

烧伤创面应用生长因子后的DNA分析

将基因重组的人表皮细胞生长因子，酸性成纤维细胞生长因子，碱性成纤维细胞生长因子和牛脑提取液外用于烧伤创面，旨在探讨皮肤组织和细胞ＤＮＡ的含量及周期变化。实验结果显示：烧伤创面经应用ｒｈ－ＥＧＦ，ｒ－ａＦＧＦ－ｒ－ｂＦＧＦ和ｂＢＥ后第５天与生理盐水组相比，创面愈合率，表皮细胞和全厚层皮肤组织的ＤＮＡ含量均高于ＮＳ组；全厚层皮肤ＤＮＡ的流式细胞仪分析表明，应用生长因子各组的Ｓ期细胞数百分比增高于ＮＳ组     

The combined effect of recombinant human epidermal growth factor and erythropoietin on full‐thickness wound healing in diabetic rat model

Diabetic wound is a chronic wound in which normal process of wound healing is interrupted. Lack of blood supply, infection and lack of functional growth factors are assumed as some of the conditions that lead to non‐healing environment. Epidermal growth factor (EGF) acts primarily to stimulate epithelial cell growth across wound. Erythropoietin (EPO) is a haematopoietic factor, which stimulates the production, differentiation and maturation of erythroid precursor cells. This study hypothesised combining these two factors, non‐healing process of diabetic wound will be compensated and eventually lead to acceleration of wound healing compared with single growth factor treatment. A total of 30 diabetic Sprague–Dawley rats were divided into three treatment groups (single treatment of rh‐EPO or rh‐EGF or combined treatment on a full‐thickness skin wound). To assess the wound healing effects of the components, the wound size and the healing time were measured in each treatment groups. The skin histology was examined by light microscopy and immunohistochemical analysis of proliferating markers was performed. The combined treatment with rh‐EPO and rh‐EGF improved full‐thickness wound significantly (P < 0·05) accelerating 50% healing time with higher expression of Ki‐67 compared with single growth factor‐treated groups. The combined treatment failed to accelerate the total healing time when compared with single growth factor treatments. However, the significant improvement were found in wound size reduction in the combined treatment group on day 4 against single growth factor‐treated groups (P < 0·05). This study demonstrated that the combined treatment of rh‐EPO and rh‐EGF improved the wound healing possibly through a synergistic action of each growth factor. This application provides further insight into combined growth factor therapy on non‐healing diabetic wounds.     

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     

Classification and treatment of chronic nonhealing wounds. Successful treatment with autologous platelet-derived wound healing factors (PDWHF)

Previous animal data showed that platelets contain growth factors that stimulate capillary endothelial migration (angiogenesis), fibroblast proliferation and migration, and collagen synthesis. This study utilized autologous platelet-derived wound healing factors (PDWHF) to treat 49 patients with chronic nonhealing cutaneous ulcers. Patients were classified on the basis of 20 clinical and wound status parameters to generate a wound severity index. Forty-nine patients--58% diabetic (20% with renal transplants); 16% with trauma, vasculitis, etc.; 14% with decubitus ulcers; and 6% each with venous stasis or arterial insufficiency--with a total of 95 wounds had received conventional wound care for an average of 198 weeks (range: 1-1820 weeks). After informed consent was obtained, patients received autologous PDWHF. Mean 100% healing time for all patients was 10.6 weeks. There was no abnormal tissue formation, keloid, or hypertrophic scarring. A multivariant analysis showed a direct correlation to 100% healing with initial wound size and the initiation of PDWHF therapy. This is the first clinical demonstration that locally acting growth factors promote healing of chronic cutaneous ulcers.     

Research of PDGF-BB gel on the wound healing of diabetic rats and its pharmacodynamics

BACKGROUND: One of the leading causes of impaired wound healing is diabetes mellitus. In diabetic patients, a minor skin wound often leads to serious complications. Many experiments had demonstrated that the expression of platelet-derived growth factor (PDGF) and its receptor was decreased in wounds of healing-impaired diabetic mice, indicating that a certain expression level of PDGF is essential for normal repair. MATERIALS AND METHODS: The diabetic rats was induced by a single i.p. injection of streptozotocin and a 1.8 cm diameter full-thickness wound was made on each side of the rat mid-back. Then the rats were randomly divided into five groups, with eight animals in each group as follows: blank control, vehicle control, 3.5 microg PDGF-BB/cm(2) treatment group, 7 microg PDGF-BB/cm(2) treatment group and 14 microg PDGF-BB/cm(2) treatment group for either 7 or 14 consecutive days after wounding. Re-epithelialization area was measured by computerized planimetry, percentage wound closure and percentage wound contraction was calculated, granulation tissue and collagen formation was assessed by Masson trichrome, cell proliferation (proliferating cell nuclear antigen staining) and angiogenesis (Factor VIII related antigen staining) was assessed by immunohistological methods. RESULTS: PDGF-BB treatment improved healing quality, enhanced angiogenesis, cell proliferation and epithelialization, and formed thicker and more highly organized collagen fiber deposition in full-thickness excisional wound of diabetic rats. The effects of topically applied PDGF-BB were dose-dependent. CONCLUSIONS: PDGF-BB is an important future clinical tool, particularly for stimulating soft tissue repair in patients with an impaired capacity for wound healing.     

Effect of Panax ginseng extract on the activity of diabetic fibroblasts in vitro

Numerous studies have demonstrated the various medicinal properties of Panax ginseng, including angiogenic, immuno‐stimulating, antimicrobial, and anti‐inflammatory activities, which can be helpful in chronic wound healing. However, a direct role for P. ginseng in chronic wound healing has not been demonstrated. The present study was designed to evaluate the effects of P. ginseng extract on diabetic fibroblasts in vitro. Human diabetic fibroblasts were cultured in the presence of Ginsenoside Rb1 (G‐Rb1), the active component in P. ginseng (10 ng/mL), and untreated diabetic fibroblasts were used as controls. Cell proliferation, collagen synthesis, the production of various growth factors (basic fibroblast growth factor [bFGF]; vascular endothelial growth factor [VEGF]; and transforming growth factor‐β1 [TGF‐β1]), and the synthesis of matrix metalloproteinase 1 (MMP‐1) and tissue inhibitor of metalloproteinases 1 (TIMP‐1) were compared using enzyme‐linked immunosorbent assay and immunofluorescence staining. Compared with the control group, G‐Rb1‐treated fibroblasts showed significantly (P < 0.05) higher levels of cell proliferation, collagen synthesis, VEGF, TGF‐β1, and TIMP‐1. However, no significant differences in bFGF and MMP‐1 levels were observed between the two groups. These results suggest that P. ginseng treatment may stimulate the wound‐healing activity of diabetic fibroblasts in vitro.     

Therapeutic role of growth factors in treating diabetic wound

Wounds in diabetic patients, especially diabetic foot ulcers, are more difficult to heal compared with normal wounds and can easily deteriorate, leading to amputation. Common treatments cannot heal diabetic wounds or control their many complications. Growth factors are found to play important roles in regulating complex diabetic wound healing. Different growth factors such as transforming growth factor beta 1, insulin-like growth factor, and vascular endothelial growth factor play different role...     

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。     

血小板源性伤口愈合因子促糖尿病大鼠伤口愈合与成纤维细 …

OBJECTIVE: The effect of platelet-derived wound healing factor (PDWHF) on wound healing in diabetic rats was studied. METHODS: Forty-four male SD rats were randomly divided into 2 groups. Thirty-two rats of experimental group accepted intraperitoneal injection of alloxan (1.5 mg/10 g body weight). Within one or two days after injection, while the blood sugar of the rats was higher than 180 mg/dl, the animal model of diabetic rat should have been established. Then a dorsal incision was given to every rat. After the addition of PDWHF (the experimental group) or bovine albumin (the control group), the incision was sutured up. Seven, ten and fourteen days after operation, the breaking strength of the wound was measured. On another hand, specimen from the wound was taken for the culture of fibroblasts. When the cultured fibroblasts have been incubated with 10% PDWHF for 4, 8 and 12 hours, the procollagen I (alpha 1) mRNA levels were examined respectively, and compared with those of control. RESULTS: Significant difference in wound breaking strength had been observed between PDWHF-treated incisions and the control on 7, 10 and 14 days after wounding (P < 0.01). Experiment in vitro demonstrated that the procollagen I (alpha 1) mRNA levels in wound fibroblasts incubated with 10% PDWHF for 4, 8 and 12 hours were 0.9, 3.7 and 2.2 folds higher than those in fibroblasts in control. CONCLUSION: It was suggested that direct stimulation of procollagen I (alpha 1) gene expression was one of the ways that PDWHF played its role in accelerating wound healing.     

Expression of growth factors in early wound healing in rat skin

Growth factors are a group of hormone-like polypeptides that have been shown to play a central role in different phases of wound healing. The expression of these growth factors in early wound healing has not been quantified, and the pattern and distribution of these growth factors in early wound healing has not been described completely. Furthermore the quantity and pattern of distribution of these growth factors have not been investigated in early wounds produced by various methods of surgical incision. Comparison of the rate of healing between the CO₂ laser wound and the scalpel wound has produced conflicting results. The present immunohistochemical study uses polyclonal antibodies specific for epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factor p (TGF-β), and basic fibroblast growth factor (bFGF) to observe the pattern and distribution of these growth factors in rat skin wound and elucidate whether there are differences in the expression of these growth factors which might account for the delayed healing of the CO₂ laser wounds compared to the scalpel as has been observed by some authors. Our results indicate that EGF, TGF-β, PDGF, and bFGF are expressed and distributed in same areas of the early skin wound. The area of expression of these growth factors was associated with presence of wound inflammatory cells and wound fibroblasts. Our study found that there were no significant differences in the expression of growth factors in the majority of time points between the CO₂ laser wounds and the scalpel wounds. © 1994 wiley-Liss, Inc.