The extracellular matrix of fascia-like tissues is a resilient network of collagenous fibers that withstand the forces of daily life. When overstretched, the matrix may tear, with serious consequences like pelvic organ prolapse (POP). Synthetic implants can provide mechanical support and evoke a host response that induces new matrix production, thus reinforcing the fascia. However, there is considerable risk of scar formation and tissue contraction which result in severe complications. Matrix producing fibroblasts are both mechanosensitive and contractile; their behavior depends on the implant's surface texture and mechanical straining. Here we investigate the effect of both in a newly-designed experimental setting. Electrospun scaffolds of Nylon and PLGA/PCL and a non-porous PLGA/PCL film were clamped like a drumhead and seeded with fibroblasts of POP patients. Upon confluency, scaffolds were cyclically strained for 24 or 72 h at 10% and 0.2 Hz, mimicking gentle breathing. Non-loading condition was control. Strained fibroblasts loosened their actin-fibers, thereby preventing myofibroblastic differentiation. Mechanical loading upregulated genes involved in matrix synthesis (collagen I, III, V and elastin), matrix remodeling (α-SMA, TGF-β1, MMP-2) and inflammation (COX-2, TNF-α, IL8, IL1-β). Collagen genes were expressed earlier under mechanical loading and the ratio of I/III collagen increased. Matrix synthesis and remodeling were stronger on the electrospun scaffolds, while inflammation was more prominent on the non-porous film. Our findings indicate that mechanical straining enhances the regenerative potential of fibroblasts for the regeneration of fascia-type tissues and limit the risk of scar tissue formation. These effects are stronger on an electrospun texture. STATEMENT OF SIGNIFICANCE: Pelvic organ prolapsed is a dysfunctional disease in female pelvic floor that can reduce the quality of life women. Currently, trans-vaginal knitted meshes are used to anatomically correct the dysfunctional tissues. However, the meshes can create sever adverse complications in some patients (e.g. chronic pain) in longer-term. As an alternative, we developed nanofibrous matrices by electrospinning based on different materials. We designed an in-vitro culture system and subjected cell-seeded matrices to cyclic mechanical loading. Results revealed that gentle straining of POP-cells on electrospun matrices, advances their regenerative potential at morphological and gene expression levels. Our findings, provide a proof-of-concept for using electrospun matrices as an alternative implant for pelvic floor repair, given that the parameters are designed efficiently and safely.

中文翻译：

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在电纺支架上对人成纤维细胞进行柔和的循环过滤可增强其再生潜力。

筋膜样组织的细胞外基质是胶原纤维的弹性网络，可承受日常生活的力量。过度拉伸时，基质可能会撕裂，并产生严重后果，如骨盆器官脱垂（POP）。合成植入物可以提供机械支撑并引起宿主反应，从而诱导新的基质产生，从而增强筋膜。然而，存在疤痕形成和组织收缩的严重风险，这会导致严重的并发症。产生基质的成纤维细胞具有机械敏感性和收缩性。它们的行为取决于植入物的表面纹理和机械应变。在这里，我们研究了两者在新设计的实验环境中的效果。尼龙和PLGA / PCL的电纺支架以及无孔PLGA / PCL薄膜像鼓皮一样被夹住，并播种POP患者的成纤维细胞。汇合后，将脚手架以10％和0.2 Hz的频率循环拉紧24或72 h，以模仿温和的呼吸。空载条件为对照。紧张的成纤维细胞会松动肌动蛋白纤维，从而阻止肌成纤维细胞分化。机械负荷上调参与基质合成（胶原I，III，V和弹性蛋白），基质重塑（α-SMA，TGF-β1，MMP-2）和炎症（COX-2，TNF-α，IL8，IL1-β）的基因）。胶原蛋白基因在机械负荷下较早表达，I / III胶原蛋白的比例增加。在电纺支架上，基质的合成和重塑作用更强，而在无孔膜上的炎症则更为明显。我们的发现表明，机械应变可增强成纤维细胞在筋膜型组织再生中的再生潜力，并限制形成疤痕组织的风险。这些效果在静电纺纹理上更强。意义声明：盆腔器官脱垂是女性骨盆底功能失调的疾病，会降低女性的生活质量。当前，经阴道编织网用于解剖上纠正功能失调的组织。但是，长期而言，网片会在某些患者中产生严重的不良并发症（例如，慢性疼痛）。作为替代方案，我们通过基于不同材料的静电纺丝开发了纳米纤维基质。我们设计了体外培养系统，并对接种了细胞的基质进行周期性的机械加载。结果表明，在静电纺丝基质上轻度拉伸POP细胞可提高其在形态学和基因表达水平上的再生潜力。我们的研究结果为使用电纺丝基质作为骨盆底修复的替代植入物提供了概念验证，因为可以有效且安全地设计参数。