Florine Emily M, Miller Rachel E, Liebesny Paul H, Mroszczyk Keri A, Lee Richard T, Patwari Parth, Grodzinsky Alan J
1 Biological Engineering, Massachusetts Institute of Technology , Cambridge, Massachusetts.
Tissue Eng Part A. 2015 Feb;21(3-4):637-46. doi: 10.1089/ten.TEA.2013.0679. Epub 2014 Oct 23.
Heparin-binding insulin-like growth factor 1 (HB-IGF-1) is a fusion protein of IGF-1 with the HB domain of heparin-binding epidermal growth factor-like growth factor. A single dose of HB-IGF-1 has been shown to bind specifically to cartilage and to promote sustained upregulation of proteoglycan synthesis in cartilage explants. Achieving strong integration between native cartilage and tissue-engineered cartilage remains challenging. We hypothesize that if a growth factor delivered by the tissue engineering scaffold could stimulate enhanced matrix synthesis by both the cells within the scaffold and the adjacent native cartilage, integration could be enhanced. In this work, we investigated methods for adsorbing HB-IGF-1 to self-assembling peptide hydrogels to deliver the growth factor to encapsulated chondrocytes and cartilage explants cultured with growth factor-loaded hydrogels. We tested multiple methods for adsorbing HB-IGF-1 in self-assembling peptide hydrogels, including adsorption prior to peptide assembly, following peptide assembly, and with/without heparan sulfate (HS, a potential linker between peptide molecules and HB-IGF-1). We found that HB-IGF-1 and HS were retained in the peptide for all tested conditions. A subset of these conditions was then studied for their ability to stimulate increased matrix production by gel-encapsulated chondrocytes and by chondrocytes within adjacent native cartilage. Adsorbing HB-IGF-1 or IGF-1 prior to peptide assembly was found to stimulate increased sulfated glycosaminoglycan per DNA and hydroxyproline content of chondrocyte-seeded hydrogels compared with basal controls at day 10. Cartilage explants cultured adjacent to functionalized hydrogels had increased proteoglycan synthesis at day 10 when HB-IGF-1 was adsorbed, but not IGF-1. We conclude that delivery of HB-IGF-1 to focal defects in cartilage using self-assembling peptide hydrogels is a promising technique that could aid cartilage repair via enhanced matrix production and integration with native tissue.
肝素结合胰岛素样生长因子1(HB-IGF-1)是胰岛素样生长因子1与肝素结合表皮生长因子样生长因子的HB结构域的融合蛋白。已证明单剂量的HB-IGF-1能特异性结合软骨,并促进软骨外植体中蛋白聚糖合成的持续上调。实现天然软骨与组织工程软骨之间的强整合仍然具有挑战性。我们假设,如果组织工程支架递送的生长因子能够刺激支架内细胞和相邻天然软骨细胞增强基质合成,那么整合可能会得到增强。在这项工作中,我们研究了将HB-IGF-1吸附到自组装肽水凝胶上的方法,以便将生长因子递送至包封的软骨细胞和与加载生长因子的水凝胶一起培养的软骨外植体。我们测试了多种将HB-IGF-1吸附到自组装肽水凝胶中的方法,包括在肽组装之前、之后吸附,以及有/无硫酸乙酰肝素(HS,肽分子与HB-IGF-1之间的潜在连接物)吸附。我们发现,在所有测试条件下,HB-IGF-1和HS都保留在肽中。然后研究了这些条件中的一部分刺激凝胶包封的软骨细胞和相邻天然软骨内软骨细胞增加基质产生的能力。与第10天的基础对照相比,发现在肽组装之前吸附HB-IGF-1或胰岛素样生长因子1能刺激软骨细胞接种水凝胶中每DNA的硫酸化糖胺聚糖和羟脯氨酸含量增加。当吸附HB-IGF-1而非胰岛素样生长因子1时,与功能化水凝胶相邻培养的软骨外植体在第10天蛋白聚糖合成增加。我们得出结论,使用自组装肽水凝胶将HB-IGF-1递送至软骨局部缺损是一种有前景的技术,可通过增强基质产生和与天然组织整合来辅助软骨修复。