Department of Orthopaedic Surgery, Hong Hui Hospital, Xi'an Jiaotong University College of Medicine, Xi'an, China.
J Orthop Surg Res. 2022 Apr 2;17(1):197. doi: 10.1186/s13018-022-03102-8.
Functionalized self-assembling peptides, which display functional growth-factor bioactivity, can be designed by connecting the C-terminus of a pure self-assembling peptide with a short functional motif. In this study, we designed a novel functionalized peptide (RADA16-SNVI) in which an SNVI motif with hBMP-7 activity was conjugated onto the C-terminus of the RADA16 peptide via solid-phase synthesis. A mix of RADA16-SNVI and RADA16 solutions was used to create a functionalized peptide nanofiber scaffold (SNVI-RADA16). The hydrogels were analyzed by atomic force microscopy, circular dichroism, and scanning electron microscopy. The results showed that the SNVI-RADA16 solution effectively formed hydrogel. Next, we seeded the SNVI-RADA16 scaffold with adipose-derived stem cells (ADSCs) and investigated whether it displayed biological properties of nucleus pulposus tissue. SNVI-RADA16 displayed good biocompatibility with the ADSCs and induced their expression. Cells in SNVI-RADA16 gel had a greater secretion of the extracellular matrix marker collagen type II and aggrecan compared to ADSCs grown in monolayer and control gel (p < 0.05). The ratio of the aggrecan to collagen in cells in SNVI-RADA16 gel is approximately 29:1 after culture for 21 days. ADSCs in SNVI-RADA16 gels expressed the hypoxia-inducible factor 1α(HIF-1α) mRNA by real-time PCR. However, HIF-1 mRNA is absence in control gel and monolayer. The results suggested that the functionalized self-assembled peptide promotes the differentiation of ADSCs into nucleus pulposus-like cells. Thus, the designed SNVI-RADA16 self-assembling peptide hydrogel scaffolds may be suitable for application in nucleus pulposus tissue regeneration.
具有功能性生长因子生物活性的功能化自组装肽可以通过将纯自组装肽的 C 末端与短功能基序连接来设计。在这项研究中,我们通过固相合成,将具有 hBMP-7 活性的 SNVI 基序连接到 RADA16 肽的 C 末端,设计了一种新型功能化肽(RADA16-SNVI)。使用 RADA16-SNVI 和 RADA16 溶液的混合物来创建功能化肽纳米纤维支架(SNVI-RADA16)。通过原子力显微镜、圆二色性和扫描电子显微镜分析水凝胶。结果表明,SNVI-RADA16 溶液有效地形成水凝胶。接下来,我们在 SNVI-RADA16 支架上接种脂肪来源的干细胞(ADSCs),并研究其是否具有核髓组织的生物学特性。SNVI-RADA16 与 ADSCs 具有良好的生物相容性,并诱导其表达。与在单层和对照凝胶中生长的 ADSCs 相比,SNVI-RADA16 凝胶中的细胞对细胞外基质标记物 II 型胶原和聚集蛋白聚糖的分泌更大(p < 0.05)。在培养 21 天后,SNVI-RADA16 凝胶中细胞的聚集蛋白聚糖与胶原的比值约为 29:1。通过实时 PCR,SNVI-RADA16 凝胶中的 ADSCs 表达缺氧诱导因子 1α(HIF-1α)mRNA。然而,对照凝胶和单层中不存在 HIF-1 mRNA。结果表明,功能化自组装肽促进 ADSCs 分化为类似髓核的细胞。因此,设计的 SNVI-RADA16 自组装肽水凝胶支架可能适用于髓核组织再生。