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Fibrin- versus Plasma-Gel Scaffolds - und der Einfluss von TGF-ß und bFGF auf Myofibroblasten und die Gewebeneogenese

机译:纤维蛋白与血浆凝胶支架-以及TGF-ß和bFGF对成肌纤维细胞和组织发生的影响

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摘要

One central aspect in tissue engineering is the scaffold. Due to its properties and the direct contact to cells, it strongly influences the development of the tissue engineered construct. Especially attractive are autologous materials, like fibrin. To optimize autologous PFP- and fibrin-scaffolds this work compared the basic materials platelet free plasma (PFP), platelet poor plasma (PPP), and platelet rich plasma (PRP) as autologous materials for scaffold production. Three differently concentrated PPP and PRP-gels were used. Therefore the microstructure of gels, the mechanical strength, proliferation, ECM-development and neogenesis were analyzed. Furthermore the influence of the growth factors bFGF and TGFß on myofibroblasts/HUASMCs was investigated.In conclusion all experiments show better results for the highly concentrated plasma-gels in contrast to the PFP/fibrin-gels. In particular, it could be shown that cells cultivated in PPP- and PRP-gels proliferated, produced collagen and migration was positively influenced also without the addition of FCS. Between PPP and PRP the differences were minor, just in comparison to PFP/fibrin-gels the difference were strong. So it seems that the key element is the resuspention of the fibrinogen pellet after cryoprecipitation in plasma. The results suggest that in follow-up studies the resuspention medium should be plasma and washing steps should be avoided to obtain a better basic material for fibrin based scaffolds. Furthermore the optimization mechanical strength and the reduction of blood volume should be investigated in upcoming studies.Myofibroblasts/HUASMCS had no receptors for TGF-ß, but for bFGF. Still no effect on mechanical strength, collagen synthesis, apoptosis, proliferation and necrosis on myofibroblasten/HUASMCs could be observed due to bFGF. More likely it is to expect that bFGF negatively influences collagen synthesis. In follow-up studies different combinations of growth factors should be investigated, because it is likely that those factors influence each other. The investigation of different combinations of growth factors could lead to the right combination for the desired effects on HUASMCs.
机译:组织工程学的一个主要方面是支架。由于其特性以及与细胞的直接接触,它强烈影响组织工程构建体的发展。诸如纤维蛋白之类的自体材料尤其有吸引力。为了优化自体PFP和纤维蛋白支架,这项工作比较了基本材料无血小板血浆(PFP),贫血小板血浆(PPP)和富血小板血浆(PRP)作为用于支架生产的自体材料。使用了三种不同浓度的PPP和PRP凝胶。因此,分析了凝胶的微观结构,机械强度,增殖,ECM发育和新生。此外,研究了生长因子bFGF和TGFβ对成肌纤维细胞/ HUASMCs的影响。总之,所有实验均表明,与PFP /纤维蛋白凝胶相比,高浓度血浆凝胶的结果更好。特别是,可以证明在不添加FCS的情况下,在PPP和PRP凝胶中培养的细胞增殖,产生胶原蛋白,并且迁移受到积极影响。 PPP和PRP之间的差异很小,只是与PFP /纤维蛋白凝胶相比,差异很大。因此看来,关键因素是血浆中冷沉淀后纤维蛋白原沉淀物的恢复。结果表明,在后续研究中,复苏介质应为血浆,应避免洗涤步骤,以获得更好的基于纤维蛋白的支架基础材料。此外,在即将进行的研究中还应研究最佳的机械强度和减少血容量。肌成纤维细胞/ HUASMCS没有TGF-ß受体,但有bFGF受体。由于bFGF,仍未观察到对肌成纤维细胞/ HUASMCs的机械强度,胶原蛋白合成,凋亡,增殖和坏死的影响。更有可能期望bFGF对胶原蛋白的合成产生负面影响。在后续研究中,应研究生长因子的不同组合,因为这些因子可能相互影响。对生长因子不同组合的研究可能会导致正确组合,从而达到对HUASMC的预期效果。

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  • 作者

    Dietrich Maren;

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  • 年度 2015
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  • 原文格式 PDF
  • 正文语种 ger
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