Vogt P M, Lehnhardt M, Wagner D, Jansen V, Krieg M, Steinau H U
Universitätsklinik für Plastische Chirurgie und Schwerbrandverletzte, Handchirurgiezentrum and Institut für Klinische Chemie und Laboratoriumsmedizin BG-Kliniken Bergmannsheil, Ruhr-Universität Bochum, Germany.
Plast Reconstr Surg. 1998 Jul;102(1):117-23. doi: 10.1097/00006534-199807000-00018.
Growth factors are important substances in the central control of wound healing during the exudative phase. Although these peptides have been applied frequently to chronic wounds in clinical studies, little is known about the naturally occurring levels at the wound site in correlation to healing in superficial wounds. We have therefore investigated the presence of these cytokines in partial thickness wounds. In 16 patients undergoing reconstructive surgery, split-thickness skin wounds were enclosed in cutaneous vinyl chambers filled with 2.5 ml of saline. Chambers placed over unwounded skin served as controls. After 24 hours, the accumulated wound fluid was harvested and replaced by 2.5 ml of saline until the wounds were healed. Wound fluid was centrifuged, aliquoted, and frozen at -70 degrees C. Samples were analyzed for protein and growth factors (insulin-like growth factor-1, epidermal growth factor, basic fibroblast growth factor, platelet-derived growth factor-AB, interleukin-1alpha, and transforming growth factor-beta1 and -beta2) and insulin-like growth factor-binding proteins 1 and 3 using a monoclonal Sandwich enzyme-linked immunosorbent assay and radioimmunoassay. All wounds healed in the liquid environment within 7 days (macroscopically) and 11 days (barrier function), respectively. In wound fluid, protein concentrations dropped from 5 mg/ml on day 1 to a baseline of 0.1 mg (unwounded skin), indicating a return of the barrier function. All growth factors could be measured already after 24 hours postwounding. However, the concentrations measured varied from 10 to more than 10,000 pg/ml between the different factors. The highest range was found for insulin-like growth factor-1 (21,000 to 41,000 pg/ml), the lowest for epidermal growth factor (3 to 63 and 3 to 88 pg/ml, respectively). Two different patterns of kinetics were distinguished: (1) a high initial peak decreasing to baseline values or below serum levels by the time of healing (insulin-like growth factor-1, insulin-like growth factor binding protein-1, -3, basic fibroblast growth factor, epidermal growth factor, platelet-derived growth factor-AB, transforming growth factor-beta1) and (2) a low initial concentration followed by an increase to a maximum at the time of epithelialization (interleukin-1alpha, transforming growth factor-beta2). Comparing the growth factor levels measured to serum baseline values, it was found that four of the growth factors appeared in wound fluid at above serum concentrations (interleukin-1alpha, transforming growth factor-beta2, basic fibroblast growth factor, epidermal growth factor); the other factors never reached serum values in wound fluid (insulin-like growth factor, transforming growth factor-beta1, platelet-derived growth factor-AB). It is concluded that the different profiles of secretion might reflect different functions of polypeptide growth factors such as stimulation of epithelialization (epidermal growth factor, insulin-like growth factor-1), matrix synthesis (transforming growth factor-beta), and inflammatory stimulation (interleukin-1alpha). The concentrations determined could serve as guidelines for adapted administration of growth factors once correlations to healing disorders such as overhealing and ulceration are established.
生长因子是渗出期伤口愈合中枢控制中的重要物质。尽管这些肽在临床研究中已频繁应用于慢性伤口，但对于浅表伤口愈合过程中伤口部位的天然存在水平却知之甚少。因此，我们研究了这些细胞因子在部分厚度伤口中的存在情况。在16例接受重建手术的患者中，将中厚皮片伤口置于充满2.5 ml生理盐水的皮肤乙烯基腔室中。置于未受伤皮肤上方的腔室作为对照。24小时后，收集积聚的伤口渗出液，并用2.5 ml生理盐水替换，直至伤口愈合。将伤口渗出液离心、分装，并在-70℃下冷冻。使用单克隆夹心酶联免疫吸附测定法和放射免疫测定法分析样品中的蛋白质和生长因子（胰岛素样生长因子-1、表皮生长因子、碱性成纤维细胞生长因子、血小板衍生生长因子-AB、白细胞介素-1α以及转化生长因子-β1和-β2）以及胰岛素样生长因子结合蛋白1和3。所有伤口在液体环境中分别于7天（宏观上）和11天（屏障功能）内愈合。在伤口渗出液中，蛋白质浓度从第1天的5 mg/ml降至0.1 mg（未受伤皮肤）的基线水平，表明屏障功能恢复。所有生长因子在受伤后24小时即可检测到。然而，不同因子测得的浓度在10至超过10,０００ pg/ml之间变化。胰岛素样生长因子-1的浓度范围最高（21,０００至41,０００ pg/ml），表皮生长因子的浓度范围最低（分别为3至63 pg/ml和3至88 pg/ml）。区分出两种不同的动力学模式：（1）一个高初始峰值，在愈合时降至基线值或低于血清水平（胰岛素样生长因子-1、胰岛素样生长因子结合蛋白-1、-3、碱性成纤维细胞生长因子、表皮生长因子、血小板衍生生长因子-AB、转化生长因子-β1）；（2）一个低初始浓度，随后在上皮化时增加至最大值（白细胞介素-1α、转化生长因子-β2）。将测得的生长因子水平与血清基线值进行比较，发现其中四种生长因子在伤口渗出液中的浓度高于血清浓度（白细胞介素-1α、转化生长因子-β2、碱性成纤维细胞生长因子、表皮生长因子）；其他因子在伤口渗出液中从未达到血清值（胰岛素样生长因子、转化生长因子-β1、血小板衍生生长因子-AB）。结论是，不同的分泌模式可能反映了多肽生长因子的不同功能，如刺激上皮化（表皮生长因子、胰岛素样生长因子-1）、基质合成（转化生长因子-β）和炎症刺激（白细胞介素-1α）。一旦建立与诸如过度愈合和溃疡等愈合障碍的相关性，所确定的浓度可作为生长因子适应性给药的指导原则。
