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Modeling early stage bone regeneration with biomimetic electrospun fibrinogen nanofibers and adipose-derived mesenchymal stem cells 用仿生电纺丝纤维蛋白原纳米纤维和脂肪来源的间充质干细胞模拟早期骨再生
Pub Date : 2016-01-04 DOI: 10.1515/esp-2016-0002
Michael P. Francis, Yas M. Moghaddam-White, Patrick C. Sachs, M. Beckman, Stephen M. Chen, G. Bowlin, L. Elmore, S. Holt
Abstract The key events of the earliest stages of bone regeneration have been described in vivo although not yet modeled in an in vitro environment, where mechanistic cell-matrix-growth factor interactions can be more effectively studied. Here, we explore an early-stage bone regeneration model where the ability of electrospun fibrinogen (Fg) nanofibers to regulate osteoblastogenesis between distinct mesenchymal stem cells populations is assessed. Electrospun scaffolds of Fg, polydioxanone (PDO), and a Fg:PDO blend were seeded with adipose-derived mesenchymal stem cells (ASCs) and grown for 7-21 days in osteogenic differentiation media or control growth media. Scaffolds were analyzed weekly for histologic and molecular evidence of osteoblastogenesis. In response to osteogenic differentiation media, ASCs seeded on the Fg scaffolds exhibit elevated expression of multiple genes associated with osteoblastogenesis. Histologic stains and scanning electron microscopy demonstrate widespread mineralization within the scaffolds, as well as de novo type I collagen synthesis. Our data demonstrates that electrospun Fg nanofibers support ASC osteogenic differentiation, yet the scaffold itself does not appear to be osteoinductive. Together, ASCs and Fg recapitulate early stages of bone regeneration ex vivo and presents a prospective autologous therapeutic approach for bone repair.
骨再生早期阶段的关键事件已经在体内描述,尽管尚未在体外环境中建模,在体外环境中可以更有效地研究细胞基质-生长因子相互作用的机制。在这里,我们探索了早期骨再生模型,其中评估了电纺纤维蛋白原(Fg)纳米纤维调节不同间充质干细胞群体之间成骨细胞发生的能力。将Fg、聚二氧环酮(PDO)和Fg:PDO混合物的电纺丝支架与脂肪来源的间充质干细胞(ASCs)相结合,在成骨分化培养基或对照生长培养基中培养7-21天。每周对支架进行成骨细胞发生的组织学和分子证据分析。在成骨分化培养基的作用下,植于Fg支架上的ASCs表现出与成骨细胞发生相关的多种基因的表达升高。组织学染色和扫描电镜显示支架内广泛的矿化,以及从头合成I型胶原蛋白。我们的数据表明,静电纺Fg纳米纤维支持ASC成骨分化,但支架本身似乎不具有成骨诱导作用。ASCs和Fg共同概括了体外骨再生的早期阶段,并为骨修复提供了一种前瞻性的自体治疗方法。
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引用次数: 5
Sub-micron sized saccharide fibres via electrospinning 通过静电纺丝制备亚微米大小的糖纤维
Pub Date : 2015-12-01 DOI: 10.1515/esp-2016-0001
Pablo G. T. Lepe, N. Tucker, Lyall D. Simmons, A. Watson, A. Fairbanks, M. Staiger
Abstract In this work, the production of continuous submicron diameter saccharide fibres is shown to be possible using the electrospinning process. The mechanism for the formation of electrospun polymer fibres is usually attributed to the physical entanglement of long molecular chains. The ability to electrospin continuous fibre from a low molecular weight saccharides was an unexpected phenomenon. The formation of sub-micron diameter “sugar syrup” fibres was observed in situ using highspeed video. The trajectory of the electrospun saccharide fibre was observed to follow that typical of electrospun polymers. Based on initial food grade glucose syrup tests, various solutions based on combinations of syrup components, i.e. mono-, di- and tri-saccharides, were investigated to map out materials and electrospinning conditions thatwould lead to the formation of fibre. Thiswork demonstrated that sucrose exhibits the highest propensity for fibre formation during electrospinning amongst the various types of saccharide solutions studied. The possibility of electrospinning low molecular weight saccharides into sub-micron fibres has implications for the electrospinability of supramolecular polymers and other biomaterials.
在这项工作中,证明了使用静电纺丝工艺生产连续亚微米直径的糖纤维是可能的。静电纺丝聚合物纤维的形成机理通常归因于长分子链的物理缠结。从低分子量的糖中静电纺连续纤维的能力是一个意想不到的现象。利用高速视频原位观察了亚微米直径“糖浆”纤维的形成。观察到电纺糖纤维的运动轨迹遵循典型的电纺聚合物。在最初的食品级葡萄糖浆测试的基础上,研究了基于糖浆成分组合的各种解决方案,即单糖、二糖和三糖,以确定导致纤维形成的材料和静电纺丝条件。这项工作表明,在研究的各种糖类溶液中,蔗糖在静电纺丝过程中表现出最高的纤维形成倾向。将低分子量糖静电纺丝成亚微米纤维的可能性对超分子聚合物和其他生物材料的电可纺丝性具有重要意义。
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引用次数: 5
Fabrication and characterization of air-impedance electrospun polydioxanone templates 空气阻抗静电纺聚二氧环酮模板的制备与表征
Pub Date : 2015-04-26 DOI: 10.1515/esp-2016-0003
Gretchen S. Selders, A. Fetz, Shannon L. Speer, G. Bowlin
Abstract Electrospinning, a fabrication technique used to create non-woven, porous templates from natural and synthetic polymers, is commonly used in tissue engineering because it is highly tailorable. However, traditional electrospinning creates restrictive pore sizes that limit the required cell migration. Therefore, tissue engineering groups have sought to enhance and regulate porosity of tissue engineering templates. Air-impedance electrospinning generates templates with tailorable, patterned areas of low and high density fiber deposition. Here we demonstrate an improved air-impedance electrospinning system, consisting of a newly designed funnel equipped to hold changeable porous deposition plates and administer air flow in a confined and focused manner, with parameters that maintain template integrity. In this preliminary study, we quantify the increase in porosity of polydioxanone templates with use of traditional fiber and pore analysis as well as with mercury porosimetry. Additionally, we validate the system’s significance in fabricating enhanced porosity templates that maintain their mechanical properties (i.e. elastic modulus, tensile strength, and suture retention strength) despite the deliberate increase in porosity. This is of exceptional value to the template’s integrity and efficacy as these parameters can be further optimized to induce the desired template porosity, strength, and texture for a given application.
静电纺丝是一种利用天然和合成聚合物制造无纺布多孔模板的制造技术,由于其高度可定制性,在组织工程中被广泛使用。然而,传统的静电纺丝产生了限制性的孔大小,限制了所需的细胞迁移。因此,组织工程小组一直在寻求提高和调节组织工程模板的孔隙率。空气阻抗静电纺丝产生具有可定制的、有图案的低密度和高密度纤维沉积区域的模板。在这里,我们展示了一种改进的空气阻抗静电纺丝系统,由一个新设计的漏斗组成,该漏斗用于容纳可变多孔沉积板,并以受限和集中的方式管理空气流动,其参数保持模板的完整性。在这项初步研究中,我们使用传统的纤维和孔隙分析以及汞孔隙度法量化了聚二氧环酮模板孔隙度的增加。此外,我们验证了系统在制造增强孔隙度模板方面的重要性,尽管孔隙度故意增加,但仍能保持其机械性能(即弹性模量、抗拉强度和缝合保持强度)。这对模板的完整性和有效性具有特殊的价值,因为这些参数可以进一步优化,以获得给定应用所需的模板孔隙率、强度和纹理。
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引用次数: 6
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Electrospinning
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