局灶性光热作用诱导胶原重建及其机制的实验研究

发布时间：2018-03-16 11:19

  本文选题：局灶性光热作用　切入点：胶原重建　出处：《上海交通大学》2012年博士论文　论文类型：学位论文

【摘要】：局灶性光热作用是一种新型的激光照射方法，通过点阵激光烧蚀部分皮肤，修复和重建皮肤表层和真皮层，达到激光美容的疗效。与传统激光美容方法相比，它减少了水肿、瘢痕、红斑或色素变化等不良反应，也克服了非剥脱性激光治疗时一次照射效果不佳的弱点。局灶性光热作用通过点阵状的微治疗孔诱发热损伤后的皮肤修复和胶原重建，这一复杂过程涉及诸多细胞和生长因子，但由于人体取材的限制，组织学发展过程还没有得到系统的定量分析，具体的调控机制也没有明确。明确局灶性光热作用下胶原重建的特征和机理，可以为点阵激光的皮肤学临床应用提供有价值的实验依据和理论基础。 针对局灶性光热作用基础研究的现状和存在问题，我们建立小鼠模型进行了系统的研究。通过对生物物理学特性的测量，首先对照比较了局灶性光热作用和传统剥脱性及非剥脱性激光在胶原重建过程中对皮肤表面结构功能的影响的异同之处；在了解这种作用模式的治疗特点后，进一步通过HE染色、双重染色、免疫组化、酶联免疫法等多种手段揭示局灶性光热作用后皮肤修复和胶原重建的组织学特点，特别定量分析了成纤维细胞增殖和胶原重建的变化情况；最终通过定量地评价和比较胶原重建过程中生长因子VEGF、TGF-β1和bFGF表达水平的变化，与皮肤组织学特征相联系，揭示局灶性光热作用诱导皮肤修复和胶原重建过程中的细胞及生长因子调控机制。 实验结果发现，相对于传统的激光治疗，局灶性光热作用诱导胶原重建过程中可有效改善皮肤弹性值，而皮肤红斑值、黑素值等不良反应表现轻微，代表皮肤屏障功能的经皮失水量和皮肤含水量都显示了安全性，从皮肤表面结构和功能上肯定了局灶性光热作用的有效性和安全性，，这项新技术临床应用前景令人期待。局灶性光热作用的皮肤组织学表现为点阵状的热损伤和可控性的修复，有效刺激了成纤维细胞和胶原的增生，早期增生胶原以Ⅲ型为主，中后期以Ⅰ型为主，总量上以Ⅰ型为主，推测局灶性光热作用后的胶原重建以Ⅰ型胶原增生为主，揭示了局灶性光热作用的胶原重建特点。最为重要的是，实验结果显示VEGF、TGF-β1和bFGF等关键生长因子在不同的修复阶段起着重要作用，推测其中VEGF在修复早期起作用，TGF-β1和bFGF在整个修复过程中起作用，TGF-β1和其他两个生长因子可能存在相互影响，它们的主要作用与刺激成纤维细胞增殖和促进胶原重建有关。 本研究系统地评价和分析了局灶性光热作用在动物模型中对皮肤组织的确切疗效和作用机制，为局灶性光热作用这一技术在皮肤科临床上的应用提供了理论基础和实验依据。
[Abstract]:Focal photothermia is a new type of laser irradiation method, which uses dot array laser ablation of part of the skin, repair and reconstruction of the skin surface and dermis to achieve the therapeutic effect of laser cosmetology. Compared with the traditional laser cosmetic method, it reduces edema. Adverse reactions, such as scar, erythema or pigment changes, also overcome the weakness of a single irradiation effect during non-stripping laser therapy. Focal photothermia induces skin repair and collagen reconstruction after thermal injury through a lattice of microtherapeutic holes. This complex process involves many cells and growth factors, but the histologic development process has not been systematically analyzed because of the limitations of the human body's selection of materials. The characteristics and mechanism of collagen reconstruction under focal photothermia can provide valuable experimental and theoretical basis for the clinical application of lattice laser dermatology. In view of the present situation and existing problems in the basic research of focal photothermal action, we have established a mouse model for systematic study, which is based on the measurement of biophysical properties. Firstly, the effects of focal photothermal action on skin surface structure and function during collagen reconstruction were compared with those of traditional exfoliation and non-stripping laser. The histological features of skin repair and collagen reconstruction after focal photothermia were revealed by HE staining, double staining, immunohistochemistry and enzyme linked immunosorbent assay (Elisa). In particular, the changes of fibroblast proliferation and collagen reconstruction were quantitatively analyzed, and the changes of the expression levels of VEGF TGF- 尾 1 and bFGF during collagen reconstruction were quantitatively evaluated and compared, which were related to the histological characteristics of skin. To explore the regulation mechanism of cell and growth factor during skin repair and collagen reconstruction induced by focal photothermia. The results showed that compared with the traditional laser therapy, focal photothermia induced collagen reconstruction could effectively improve the skin elasticity, while the skin erythema value, melanin value and other adverse reactions showed slight. Both skin water loss and skin moisture content, which represent skin barrier function, are safe. The effectiveness and safety of focal photothermal action are confirmed from skin surface structure and function. The prospect of clinical application of this new technique is promising. The skin histology of focal photothermia is characterized by lattice thermal damage and controllable repair, which effectively stimulates the proliferation of fibroblasts and collagen. In the middle and late stage, type I was the main type, and the total amount was type 鈪

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