Department of Biomedical Engineering, Washington University in St. Louis, USA.
Department of Orthopedic Surgery, Washington University School of Medicine, USA.
Biomaterials. 2021 Oct;277:121113. doi: 10.1016/j.biomaterials.2021.121113. Epub 2021 Sep 1.
Biomaterial based strategies have been widely explored to preserve and restore the juvenile phenotype of cells of the nucleus pulposus (NP) in degenerated intervertebral discs (IVD). With aging and maturation, NP cells lose their ability to produce necessary extracellular matrix and proteoglycans, accelerating disc degeneration. Previous studies have shown that integrin or syndecan binding peptide motifs from laminin can induce NP cells from degenerative human discs to re-express juvenile NP-specific cell phenotype and biosynthetic activity. Here, we engineered alginate hydrogels to present integrin- and syndecan-binding peptides alone or in combination (cyclic RGD and AG73, respectively) to introduce bioactive features into the alginate gels. We demonstrated human NP cells cultured upon and within alginate hydrogels presented with cRGD and AG73 peptides exhibited higher cell viability, biosynthetic activity, and NP-specific protein expression over alginate alone. Moreover, the combination of the two peptide motifs elicited markers of the NP-specific cell phenotype, including N-Cadherin, despite differences in cell morphology and multicellular cluster formation between 2D and 3D cultures. These results represent a promising step toward understanding how distinct adhesive peptides can be combined to guide NP cell fate. In the future, these insights may be useful to rationally design hydrogels for NP cell-transplantation based therapies for IVD degeneration.
生物材料策略已被广泛探索用于保存和恢复退化椎间盘核髓核(NP)细胞的幼年表型。随着年龄的增长和成熟,NP 细胞丧失了产生必需的细胞外基质和蛋白聚糖的能力,加速了椎间盘的退化。先前的研究表明,来自层粘连蛋白的整联蛋白或连接蛋白结合肽基序可以诱导来自退行性人椎间盘的 NP 细胞重新表达幼年 NP 特异性细胞表型和生物合成活性。在这里,我们设计了藻酸盐水凝胶来单独或组合呈现整联蛋白和连接蛋白结合肽(分别为环状 RGD 和 AG73),以将生物活性特征引入藻酸盐凝胶中。我们证明,在藻酸盐水凝胶上和内部培养的人 NP 细胞表现出更高的细胞活力、生物合成活性和 NP 特异性蛋白表达,而单独使用藻酸盐时则没有。此外,尽管 2D 和 3D 培养之间的细胞形态和多细胞簇形成存在差异,但两种肽基序的组合引发了 NP 特异性细胞表型的标志物,包括 N-钙粘蛋白。这些结果代表了理解如何结合不同的粘附肽来指导 NP 细胞命运的一个有希望的步骤。将来,这些见解可能有助于合理设计用于椎间盘退行性变的 NP 细胞移植治疗的水凝胶。