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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 玛丽·C·法拉赫-卡森 电纺胶原支架用于抗肿瘤药物三维细胞模型的研究 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。这反映了这样一个事实，即这些表面细胞首当其冲地承受着活细胞受到包括物理和化学刺激在内的环境侮辱的影响。当这些细胞从正常细胞转化为癌细胞时，它们的性质就会改变。肿瘤是由细胞从其自然衬里(或基底膜)中释放出来形成的，并形成相互作用的三维结构，并与肿瘤周围组织的微环境相互作用。在塑料组织培养上生长平坦的癌细胞作为单层并不能反映整个肿瘤的许多特性。这一缺陷限制了它们作为测试药理活性化合物的完美模型的能力，包括那些正在作为抗癌药物(抗新塑料)进行测试的化合物。我们建议将我们在生物学(BIO)和材料科学与工程(MSE)的单独实验室中优化的两项技术结合在一起，创造出具有与癌症周围天然组织中更相似的特性的新的3-D细胞材料。这项工作的目标是生产一种静电纺丝的微米和纳米纤维支架，将支持肿瘤在三维空间的生长。电纺是电喷雾的一个分支，将被用于纺制蜘蛛网类型的纤维，细胞将在这些纤维上生长，用于表征和测试抗癌化合物。在静电纺丝过程中产生的纤维是纳米级的，与直径为5-200米的传统纺织纤维相比，直径在40-2000纳米之间。静电纺丝的主要优势是它在溶液中使用微量(50-100毫克，可能是定制合成的量)来形成微米和纳米纤维。第二个优点是，可以将例如小分子、第二聚合物或细胞结合因子的附加成分添加到聚合物溶液中，并且通常在电纺过程中将其并入纤维中。在可行性研究中，选择I型胶原作为基质材料是因为它是天然纤维的主要成分，因此在结构上可以模拟天然细胞外基质(ECM)的物理环境。胶原蛋白本身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的蛋白质表现出很高的亲和力。我们计划用ECM基底膜蛋白的小段重组片段覆盖胶原基支架，我们已经证明这是一种有用的聚乳酸(PLA)支架上的蛋白质涂层材料。我们相信，这种涂层将为癌细胞提供更自然的环境，使它们的生长方式与人类肿瘤更加相似。因此，它将提供一种更好的方法来测试癌细胞对药理活性化合物的反应，并将为在3-D培养中测试潜在的抗癌新药提供一个更好的模型。

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.