A Bio-manufacturing process for fabrication of 3D scaffolds with cell culture der

描述(申请人提供)：本研究的目标是开发具有仿生细胞外基质(ECM)涂层的新型3D可生物降解多孔聚合物支架，用于组织工程应用。这项SBIR第一阶段的建议是通过使用专利的精密挤压沉积技术来制造支架的可行性，以制造具有可控孔隙率和内部结构的多孔3D组织支架，然后利用细胞培养技术在支架上覆盖细胞培养来源的细胞外基质(ECM)。我们推测，这种细胞培养来源的仿生ECM涂层经过适当的脱细胞和处理后，在组织工程应用中可以提供与传统组织来源ECM相似的生物学性能。为了实现这一目标，我们想要证明可以在多孔组织支架的表面生成一层ECM涂层。为了实现这一目标，我们首先应用了一种能够实现精密挤出沉积的技术来自由地制备具有不同孔道结构的多孔3D聚己内酯(PCL)支架。然后，我们应用细胞培养方法，用人真皮成纤维细胞和人间充质干细胞(在成骨分化情况下)制备了两种类型的ECM涂层。我们希望进一步证明来自两种不同细胞系(成纤维细胞和成骨分化的间充质干细胞)的脱细胞仿生ECM涂层将影响重新种植的间充质干细胞的成骨分化。更具体地说，我们想要证明，从细胞附着、增殖和分化到成骨细胞谱系，从细胞附着、增殖和向下分化到成骨谱系，从细胞黏附、增殖和向下分化到成骨细胞，ECM涂层可以作为更好的ECM基质用于重新种植间充质干细胞。该项目旨在开发具有仿生细胞外基质(ECM)涂层的新型3D可生物降解多孔聚合物支架，用于组织工程应用。在这项SBIR第一阶段的工作中，我们将展示支架制造的可行性，方法是使用专利制造技术来生产孔隙率和内部结构可控的多孔3D组织支架，然后使用细胞培养技术生成细胞外基质(ECM)作为仿生涂层。我们相信，这种新的制造工艺将生产出具有理想的生物物理性能和生物性能的支架。

DESCRIPTION (provided by applicant): The objective of this research is to develop novel 3D biodegradable porous polymer scaffolds with biomimetic extracellular matrix (ECM) coating for tissue engineering application. This SBIR Phase I proposal is to demonstrate the scaffold manufacturing feasibility by using a proprietary precision extruding deposition technique to fabricate porous 3D tissue scaffold with controlled porosity and internal architecture, and then coated the scaffold with a cell culture derived extracellular matrix (ECM) by using cell culture technology. We hypotheze that this cell culture derived biomimetic ECM coating, after proper decellularization and treatment, can offer similar biological performance as conventional tissue derived ECM in tissue engineering application. Toward this goal, we would like to demonstrate that an ECM coating can be generated onto the surface of porous tissue scaffolds. To achieve this aim, we first apply an enabling precision extruding deposition technique to freeformly fabricate porous 3D polycaprolactone (PCL) scaffolds with various porous structures. We then apply cell culture method to produce two types of ECM coatings using human dermal fibroblasts and human mesenchymal stem cells (under osteogeneic differentiation). We would like to further demonstrate that the decellularized biomimetic ECM coatings derived from two different cell lines (fibroblast and mesenchymal stem cells under osteogeneic differentiation) will affect osteo-differentiation of re-seeded mesenchymal stem cells. More specifically, we would like to demonstrate that the osteo- differentiated mesenchymal stem cells derived ECM coating could be used as a better ECM substrate for re-seeded mesenchymal stem cell in terms of the cell attachment, proliferation and differentiation down to the osteogeneic lineage. PUBLIC HEALTH RELEVANCE The project is aimed at developing novel 3D biodegradable porous polymer scaffolds with biomimetic extracellular matrix (ECM) coating for tissue engineering application. In this SBIR Phase I work, we will demonstrate the scaffold manufacturing feasibility by using a proprietary fabrication technique to produce porous 3D tissue scaffold with controlled porosity and internal architecture, and then use a cell culture technique to generate an extracellular matrix (ECM) as biomimetic coating. We believe that this novel manufacturing process will produce scaffolds with desired both biophysical properties and biological performance.