Enhanced trophic factor secretion by mesenchymal stem/stromal cells with Glycine-Histidine-Lysine (GHK)-modified alginate hydrogels.
Enhanced trophic factor secretion by mesenchymal stem/stromal cells with Glycine-Histidine-Lysine (GHK)-modified alginate hydrogels.
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DOI：
10.1016/j.actbio.2014.01.020
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发表时间：
2014-05
影响因子：
9.7
通讯作者：
Leach, J. Kent
中科院分区：
文献类型：
--
作者：
Jose, Soumia;Hughbanks, Marissa L.;Binder, Bernard Y. K.;Ingavle, Ganesh C.;Leach, J. Kent
重组蛋白和细胞因子正在进行广泛的临床前和临床研究，以促进血管生成，但它们的成功受到输送效率低、缺乏长期稳定性和成本过高的限制。间充质干细胞/基质细胞(MSC)分泌具有生物活性的营养因子，可能为解决这些挑战提供一种有效的替代方法。甘氨酸-组氨酸-赖氨酸(GHK)是骨连接蛋白(SPARC)的多肽片段，是一种具有促血管生成潜能的基质细胞蛋白。我们检测了GHK在培养和共价偶联到藻酸盐水凝胶上时，上调人MSC分泌促血管生成因子的能力。GHK在较宽的浓度范围内对培养的MSC无明显的细胞毒作用。我们发现在经GHK处理的MSC条件下，培养上清液中血管内皮生长因子(VEGF)浓度呈剂量依赖性增加，从而促进内皮细胞的增殖、迁移和小管形成。我们用碳二亚胺化学方法将GHK共价偶联到藻酸盐上，并将人的MSC包埋在藻酸盐水凝胶中，以评估血管内皮生长因子的分泌。与单层培养相似，MSC对GHK修饰凝胶的反应是分泌比未修饰凝胶更高浓度的血管内皮生长因子和碱性成纤维细胞生长因子。α-6和β-1整合素抗体可抑制血管生成，提示其促血管生成反应是由整合素介导的。这些数据表明，将GHK作为可生物降解的载体，可以显著增加MSC的促血管生成潜能，从而增加其用于组织修复的临床潜力。
Recombinant proteins and cytokines are under broad preclinical and clinical investigation to promote angiogenesis, but their success is limited by ineffective delivery, lack of long-term stability, and excessive cost. Mesenchymal stem/stromal cells (MSC) secrete bioactive trophic factors, and thus, may provide an effective alternative to address these challenges. Glycine-Histidine-Lysine (GHK) is a peptide fragment of osteonectin (SPARC), a matricellular protein with reported proangiogenic potential. We examined the capacity of GHK to upregulate secretion of proangiogenic factors from human MSC in culture and when covalently coupled to alginate hydrogels. GHK had no apparent cytotoxic effects on MSC in culture over a wide range of concentrations. We detected a dose-dependent increase in vascular endothelial growth factor (VEGF) concentration in media conditioned by GHK-treated MSC, which increased endothelial cell proliferation, migration, and tubule formation. We covalently coupled GHK to alginate using carbodiimide chemistry, and human MSC were entrapped in alginate hydrogels to assess VEGF secretion. Similar to monolayer culture, MSC responded to GHK-modified gels by secreting increased concentrations of VEGF and basic fibroblast growth factor (bFGF) compared to unmodified gels. The pre-treatment of MSC with antibodies to α6 and β1 integrins prior to entrapment in GHK-modified gels abrogated VEGF secretion, suggesting that the proangiogenic response of MSC was integrin-mediated. These data demonstrate that the proangiogenic potential of MSC can be significantly increased by the presentation of GHK with a biodegradable carrier, therefore increasing their clinical potential when used for tissue repair.
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影响因子：
23.9
作者：
Caplan AI;Correa D
通讯作者：
Correa D
影响因子：
7.6
作者：
Alsberg, E;Kong, HJ;Mooney, DJ
通讯作者：
Mooney, DJ
影响因子：
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作者：
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影响因子：
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作者：
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DOI：
10.1073/pnas.192291499
发表时间：
2002-09-17
影响因子：
11.1
作者：
Alsberg, E;Anderson, KW;Mooney, DJ
通讯作者：
Mooney, DJ