Department of Scientific Research, Hong Hui Hospital, Xi'an Jiaotong University College of Medicine, Xi'an, Shaanxi 710054, China.
Chin Med J (Engl). 2013 Oct;126(20):3891-6.
Olfactory ensheathing cell (OEC) transplantation is a promising or potential therapy for spinal cord injury (SCI). However, the effects of injecting OECs directly into SCI site have been limited and unsatisfied due to the complexity of SCI. To improve the outcome, proper biomaterials are thought to be helpful since these materials would allow the cells to grow three-dimensionally and guide cell migration.
In this study, we made a new peptide hydrogel scaffold named GRGDSPmx by mixing the pure RADA16 and designer peptide RADA16-GRGDSP solution, and we examined the molecular integration of the mixed nanofiber scaffolds using atomic force microscopy. In addition, we have studied the behavior of OECs in GRGDSPmx condition as well as on RADA16 scaffold by analyzing their phenotypes including cell proliferation, apoptosis, survival, and morphology.
The experimental results showed that GRGDSPmx could be self-assembled to form a hydrogel. Inverted optical microscopic and scanning electron microscopic analyses showed that OECs are viable and they proliferate within the nanostructured environment of the scaffold. Thiazolyl blue (MTT) assay demonstrated that OEC proliferation rate was increased on GRGDSPmx scaffold compared with the pure RADA16 scaffold. In addition, OECs on GRGDSPmx scaffolds also showed less apoptosis and maintained the original spindle-shaped morphology. Calcein-AM/PI fluorescence staining revealed that OECs cultured on GRGDSPmx grew well and the viable cell count was 95%.
These results suggested that this new hydrogel scaffold provided an ideal substrate for OEC three-dimensional culture and suggested its further application for SCI repair.
嗅鞘细胞（OEC）移植是一种有前途的脊髓损伤（SCI）治疗方法。然而，由于 SCI 的复杂性，将 OEC 直接注射到 SCI 部位的效果有限且不理想。为了改善结果，人们认为适当的生物材料是有帮助的，因为这些材料可以使细胞进行三维生长并引导细胞迁移。
在这项研究中，我们通过混合纯 RADA16 和设计肽 RADA16-GRGDSP 溶液，制作了一种名为 GRGDSPmx 的新型肽水凝胶支架，并使用原子力显微镜检查了混合纳米纤维支架的分子整合情况。此外，我们通过分析 OEC 在 GRGDSPmx 条件和 RADA16 支架上的表型，包括细胞增殖、凋亡、存活和形态，研究了 OEC 的行为。
实验结果表明，GRGDSPmx 可以自组装形成水凝胶。倒置光学显微镜和扫描电子显微镜分析表明，OEC 在支架的纳米结构环境中是存活的并且增殖。噻唑蓝（MTT）测定表明，与纯 RADA16 支架相比，OEC 的增殖率在 GRGDSPmx 支架上增加。此外，GRGDSPmx 支架上的 OEC 凋亡较少，保持原始纺锤形形态。钙黄绿素 AM/PI 荧光染色显示，在 GRGDSPmx 支架上培养的 OEC 生长良好，活细胞计数为 95%。
这些结果表明，这种新型水凝胶支架为 OEC 的三维培养提供了理想的基质，并为 SCI 修复的进一步应用提供了依据。
