ELECTROSPUN COLLAGEN SCAFFOLDS FOR 3D CELLULAR MODELS FOR ANTI-NEOPLASTIC AGENTS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 玛丽C.法拉赫-卡森 静电纺丝胶原支架用于抗肿瘤药物三维细胞模型的建立 超过90%的癌症，包括乳腺癌和前列腺癌，起源于排列在人体组织表面的上皮细胞。这反映了这些表面细胞承受活细胞暴露于包括物理和化学刺激的环境损害的冲击的事实。当这些细胞从正常细胞转化为癌细胞时，它们的性质发生了变化。肿瘤由从其天然衬里（或基底膜）释放的细胞形成，并形成彼此相互作用并与肿瘤周围组织的微环境相互作用的3-D结构。在塑料组织培养物上生长的癌细胞作为单层并不能反映整个肿瘤的许多特性。这一缺点限制了它们作为测试生物活性化合物的完美模型的能力，包括那些作为抗癌药物（抗肿瘤）进行测试的化合物。我们建议将两种技术联合收割机结合起来，这两种技术已经在我们的生物学（BIO）和材料科学与工程（MSE）实验室中进行了优化，以创建新的3-D细胞材料，这些材料具有与癌症周围天然组织更相似的特性。这项工作的目标是生产一种静电纺丝的微纤维和纳米纤维支架，以支持肿瘤的三维生长。静电纺丝是电喷雾的一个分支，将用于纺制蜘蛛网型纤维，细胞将在其上生长，用于抗癌化合物的表征和测试。在静电纺丝过程中产生的纤维是纳米级的，与直径为5 - 200 μ m的传统纺织纤维相比，直径为40 - 2000 nm。 静电纺丝的主要优点是它使用极少量（50 - 100 mg  可能由定制合成产生的量）在溶液中聚合物以形成微米和纳米纤维。 第二个优点是附加的部件，可以将小分子、第二聚合物或细胞结合因子添加到聚合物溶液中，并且通常在静电纺丝过程中将其掺入纤维中。对于可行性研究，选择胶原蛋白（I型）作为基质材料，因为它是天然纤维的主要成分，因此可以在结构上模拟天然细胞外基质（ECM）的物理环境。 单独的胶原蛋白已被证明可以促进细胞识别，并对细胞表面结合和生长因子中发现的蛋白质表现出高亲和力。我们计划用ECM基底膜蛋白的小重组片段涂覆胶原基支架，我们已经证明ECM基底膜蛋白是聚乳酸（PLA）支架上有用的蛋白质涂层材料。我们相信这种涂层将为癌细胞提供一个更自然的环境，使它们的生长更类似于人类肿瘤。因此，它将提供一种上级的方法来测试癌细胞如何对抗癌活性化合物作出反应，并将提供一种上级的模型来测试在3-D培养中的潜在的新抗癌药物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Mary C. Farach-Carson Electrospun Collagen Scaffolds for Development of 3-D Cellular Models for Testing Anti-Neoplastic Agents Greater than 90% of cancers, including those from breast and prostate, originate from epithelial cells that line the surfaces of human tissues. This reflects the fact that these surface cells bear the brunt of exposure of living cells to environmental insult including physical and chemical stimuli. As these cells are transformed from normal cells to cancer cells, their properties change. Tumors form from cells that are released from their natural lining (or basement membrane) and form 3-D structures that interact with each other and with the microenvironment of the tissue around the tumor. Cancer cells growing flat on plastic tissue culture as single layers do not reflect many of the properties of whole tumors. This shortcoming limits their ability to serve as perfect models for testing of pharmacologically active compounds, including those that are being tested as anti-cancer drugs (anti-neoplastics). We propose to combine two technologies that have been optimized in our separate laboratories in Biology (BIO) and Materials Science and Engineering (MSE) to create new 3-D cellular materials possessing properties more similar to those in native tissues surrounding cancers. The goal of this work is to produce an electrospun micro- and nanofibrous scaffold that will support tumor growth in three dimensions. Electrospinning, an offshoot of electrospraying, will be used to spin spider web type fibers on which cells will be grown for characterization and testing of anti-cancer compounds. The fibers produced during the electrospinning process are nanoscale, with diameters ranging from 40 to 2000 nm compared to traditional textile fibers that have diameters of 5-200 ¿m. The primary advantage of electrospinning is that it uses tiny quantities (50-100 mg  the quantity that might result from a custom synthesis) of polymer in solution to form micro- and nanofibers. A second advantage is that additional components, e.g., small molecules, a second polymer, or cell binding factors can be added to the polymer solution and often be incorporated into the fiber during the electrospinning process. For a feasibility study, collagen (type I) was chosen as the matrix material because it is a major constituent of natural fibers and thus can structurally mimic the physical environment of the natural extracellular matrix (ECM). Collagen alone has been shown to promote cellular recognition and exhibits a high affinity for proteins like those found in cell surface binding and growth factors. We plan to coat the collagen based scaffolds with small recombinant fragments of the ECM basement membrane proteins which we have shown to be a useful protein coating material on polylactic acid (PLA) scaffolds. We believe this coating will provide a more natural environment to cancer cells such that they will grow more similarly to human tumors. As such, it will provide a superior way to test how cancer cells respond to pharmacologically active compounds and will provide a superior model for testing potential new anti-cancer drugs in 3-D culture.