Synthetic Heparan Sulfate Hydrogels Regulate Neurotrophic Factor Signaling and Neuronal Network Activity.
Synthetic Heparan Sulfate Hydrogels Regulate Neurotrophic Factor Signaling and Neuronal Network Activity.
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DOI：
10.1021/acsami.2c01575
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发表时间：
2022-06-29
影响因子：
9.5
通讯作者：
Karumbaiah, Lohitash
中科院分区：
文献类型：
--
作者：
Latchoumane, Charles-Francois, V;Chopra, Pradeep;Sun, Lifeng;Ahmed, Aws;Palmieri, Francesco;Wu, Hsueh-Fu;Guerreso, Rebecca;Thorne, Kristen;Zeltner, Nadja;Boons, Geert-Jan;Karumbaiah, Lohitash
三维（3D）合成硫酸乙酰肝素（HS）构建体在神经组织工程应用中具有良好的特性。然而，它们依赖硫酸化促进分子识别和细胞信号传导的能力尚未得到研究。我们假设，与未硫酸化的对照构建体（含化合物2）相比，用神经黏附肽功能化的完全硫酸化合成HS构建体（含化合物1）将增强成纤维细胞生长因子 - 2（FGF2）与FGF受体 - 1（FGFR1）的结合及复合，从而促进人神经干细胞（hNSCs）的增殖和神经元分化。我们使用由HS四糖（化合物3和4）以及一种工程化整合素结合嵌合肽（CP）的不同组合构成的2D和3D基质在体外测试了这一假设，这些物质是利用应变促进的炔 - 叠氮环加成（SPAAC）化学方法组装的。结果表明，当在嵌合肽功能化的2D玻璃基质上培养时，hNSCs的黏附显著增加。与在2 - CP水凝胶中的hNSCs相比，包封在1 - CP水凝胶中并在含有有丝分裂原FGF2的培养基中培养的hNSCs表现出显著更高的神经元分化。这些观察结果通过蛋白质印迹分析得到了证实，该分析表明化合物1对FGF2和FGFR1的结合和保留增强，以及细胞外信号调节激酶（ERK1/2）的下游磷酸化和hNSCs增殖增强。最后，钙活动成像显示1和2水凝胶都支持预分化的人前额叶皮质神经元的神经元生长和活动。总之，这些结果表明合成的HS水凝胶可被定制以调节生长因子信号传导以及神经元的命运、活动。
Three-dimensional (3D) synthetic heparan sulfate (HS) constructs possess promising attributes for neural tissue engineering applications. However, their sulfation-dependent ability to facilitate molecular recognition and cell signaling has not yet been investigated. We hypothesized that fully sulfated synthetic HS constructs (bearing compound 1) that are functionalized with neural adhesion peptides will enhance fibroblast growth factor-2 (FGF2) binding and complexation with FGF receptor-1 (FGFR1) to promote the proliferation and neuronal differentiation of human neural stem cells (hNSCs) when compared to constructs with unsulfated controls (bearing compound 2). We tested this hypothesis in vitro using 2D and 3D substrates consisting of different combinations of HS tetrasaccharides (compounds 3 and 4) and an engineered integrin-binding chimeric peptide (CP), which were assembled using strain-promoted alkyne-azide cycloaddition (SPAAC) chemistry. Results indicated that the adhesion of hNSCs increased significantly when cultured on 2D glass substrates functionalized with chimeric peptide. hNSCs encapsulated in 1-CP hydrogels and cultured in media containing the mitogen FGF2 exhibited significantly higher neuronal differentiation when compared to hNSCs in 2-CP hydrogels. These observations were corroborated by Western blot analysis, which indicated the enhanced binding and retention of both FGF2 and FGFR1 by 1 as well as downstream phosphorylation of extracellular signal-regulated kinases (ERK1/2) and enhanced proliferation of hNSCs. Lastly, calcium activity imaging revealed that both 1 and 2 hydrogels supported the neuronal growth and activity of pre-differentiated human prefrontal cortex neurons. Collectively, these results demonstrate that synthetic HS hydrogels can be tailored to regulate growth factor signaling and neuronal fate and activity.
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DOI：
10.1534/g3.114.012591
发表时间：
2014-08-05
期刊：
G3 (Bethesda, Md.)
影响因子：
--
作者：
Díaz-Balzac CA;Lázaro-Peña MI;Tecle E;Gomez N;Bülow HE
通讯作者：
Bülow HE
DOI：
10.1016/j.bbagen.2008.09.001
发表时间：
2009-01-01
影响因子：
3
作者：
Asada, Masahiro;Shinomiya, Michiyo;Imamura, Toru
通讯作者：
Imamura, Toru
影响因子：
4
作者：
Chopra, Pradeep;Logun, Meghan T.;Boons, Geert-Jan
通讯作者：
Boons, Geert-Jan
影响因子：
4
作者：
Martin, VV;Beierlein, M;Gee, KR
通讯作者：
Gee, KR
影响因子：
15
作者：
Arungundram, Sailaja;Al-Mafraji, Kanar;Asong, Jinkeng;Leach, Franklin E., III;Amster, I. Jonathan;Venot, Andre;Turnbull, Jeremy E.;Boons, Geert-Jan
通讯作者：
Boons, Geert-Jan