3B's Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, 4806-909 Taipas, Guimarães, Portugal.
Acta Biomater. 2012 Apr;8(4):1530-42. doi: 10.1016/j.actbio.2011.12.015. Epub 2011 Dec 16.
An important challenge in the biomaterials field is to mimic the structure of functional tissues via cell and extracellular matrix (ECM) alignment and anisotropy. Toward this goal, silk-based scaffolds resembling bone lamellar structure were developed using a freeze-drying technique. The structure could be controlled directly by solute concentration and freezing parameters, resulting in lamellar scaffolds with regular morphology. Different post-treatments, such as methanol, water annealing and steam sterilization, were investigated to induce water stability. The resulting structures exhibited significant differences in terms of morphological integrity, structure and mechanical properties. The lamellar thicknesses were ∼2.6 μm for the methanol-treated scaffolds and ∼5.8 μm for water-annealed. These values are in the range of those reported for human lamellar bone. Human bone marrow-derived mesenchymal stem cells (hMSC) were seeded on these silk fibroin lamellar scaffolds and grown under osteogenic conditions to assess the effect of the microstructure on cell behavior. Collagen in the newly deposited ECM was found aligned along the lamellar architectures. In the case of methanol-treated lamellar structures, the hMSC were able to migrate into the interior of the scaffolds, producing a multilamellar hybrid construct. The present morphology constitutes a useful pattern onto which hMSC cells attach and proliferate for guided formation of a highly oriented extracellular matrix.
在生物材料领域,一个重要的挑战是通过细胞和细胞外基质(ECM)的排列和各向异性来模拟功能性组织的结构。为了实现这一目标,使用冷冻干燥技术开发了类似于骨板层结构的基于丝的支架。通过溶质浓度和冷冻参数可以直接控制结构,从而得到具有规则形态的层状支架。研究了不同的后处理方法,如甲醇、水退火和蒸汽灭菌,以诱导水稳定性。所得结构在形态完整性、结构和机械性能方面表现出显著差异。甲醇处理的支架的层厚约为 2.6μm,水退火的支架的层厚约为 5.8μm。这些值在人板层骨报道的范围内。将人骨髓间充质干细胞(hMSC)接种在这些丝素蛋白层状支架上,并在成骨条件下培养,以评估微观结构对细胞行为的影响。新沉积的 ECM 中的胶原蛋白被发现沿着层状结构排列。在甲醇处理的层状结构的情况下,hMSC 能够迁移到支架的内部,形成多层杂交结构。目前的形态构成了一个有用的模式,hMSC 细胞附着并增殖,以引导高度定向的细胞外基质的形成。