Tendon-derived matrix crosslinking techniques for electrospun multi-layered scaffolds.
Tendon-derived matrix crosslinking techniques for electrospun multi-layered scaffolds.
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DOI：
10.1002/jbm.a.37588
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发表时间：
2023-07
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影响因子：
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通讯作者：
Thomas L Jenkins;Paula A Sarmiento Huertas;Kentaro Umemori;F. Guilak;D. Little
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作者：
Thomas L Jenkins;Paula A Sarmiento Huertas;Kentaro Umemori;F. Guilak;D. Little
肌腱撕裂是常见的，愈合往往发生不完全和纤维化。组织工程旨在改善修复，其中一种正在研究的方法是使用含有仿生因子的细胞接种支架。仿生因子在支架上的保留对于最大化其益处，同时最小化不良反应的风险，并且不损失仿生因子的有益作用可能是至关重要的。本研究的目的是评估交联方法，以提高保留肌腱衍生基质（TDM）的静电纺丝聚（ε-己内酯）（PCL）支架。我们测试了紫外线（UV）或碳二亚胺（EDC：NHS：COOH）交联方法的效果，以更好地将TDM保留在支架上并刺激肌腱样基质合成。最初，我们测试了与未交联的TDM相比，在PCL膜上的碳二亚胺（2.5：1：1、5：2：1和10：4：1的EDC：NHS：COOH比率）和UV（30 s 1 J/cm 2、60 s 1 J/cm 2和60 s 4 J/cm 2）的各种交联构型。我们发现，没有测试的交联比单独的涂层保留更多的TDM（Kruskal-Wallis：p > 0.05），但是人脂肪干细胞（hASC）在60 μ s 1 J/cm 2 UV-和2.5：1：1 EDC-交联的膜上扩散最多（Kruskal-Wallis：p <0.05）。接下来，我们比较了60 s 1 J/cm 2 UV-和2.5：1：1 EDC-交联到TDM-涂覆的和未处理的PCL支架对hASC-诱导的肌腱样分化的影响。UV交联的支架具有比PCL或TDM支架更大的模量和刚度，并且hASC在UV交联的支架上扩展得更多（ANOVA：p < .05）。傅立叶变换红外光谱表明，UV或EDC交联的TDM不影响肌腱特征波数处的峰。将TDM交联到静电纺丝支架上改善了肌腱样基质的合成，为改善TDM在静电纺丝PCL支架上的保留提供了可行的策略。
Tendon tears are common and healing often occurs incompletely and by fibrosis. Tissue engineering seeks to improve repair, and one approach under investigation uses cell-seeded scaffolds containing biomimetic factors. Retention of biomimetic factors on the scaffolds is likely critical to maximize their benefit, while minimizing the risk of adverse effects, and without losing the beneficial effects of the biomimetic factors. The aim of the current study was to evaluate cross-linking methods to enhance the retention of tendon-derived matrix (TDM) on electrospun poly(ε-caprolactone) (PCL) scaffolds. We tested the effects of ultraviolet (UV) or carbodiimide (EDC:NHS:COOH) crosslinking methods to better retain TDM to the scaffolds and stimulate tendon-like matrix synthesis. Initially, we tested various crosslinking configurations of carbodiimide (2.5:1:1, 5:2:1, and 10:4:1 EDC:NHS:COOH ratios) and UV (30 s 1 J/cm2 , 60 s 1 J/cm2 , and 60 s 4 J/cm2 ) on PCL films compared to un-crosslinked TDM. We found that no crosslinking tested retained more TDM than coating alone (Kruskal-Wallis: p > .05), but that human adipose stem cells (hASCs) spread most on the 60 s 1 J/cm2 UV- and 2.5:1:1 EDC-crosslinked films (Kruskal-Wallis: p < .05). Next, we compared the effects of 60 s 1 J/cm2 UV- and 2.5:1:1 EDC-crosslinked to TDM-coated and untreated PCL scaffolds on hASC-induced tendon-like differentiation. UV-crosslinked scaffolds had greater modulus and stiffness than PCL or TDM scaffolds, and hASCs spread more on UV-crosslinked scaffolds (ANOVA: p < .05). Fourier transform infrared spectra revealed that UV- or EDC-crosslinking TDM did not affect the peaks at wavenumbers characteristic of tendon. Crosslinking TDM to electrospun scaffolds improves tendon-like matrix synthesis, providing a viable strategy for improving retention of TDM on electrospun PCL scaffolds.