Chen Xue, Xu Mei-Ling, Wang Cheng-Niu, Zhang Lu-Zhong, Zhao Ya-Hong, Zhu Chang-Lai, Chen Ying, Wu Jian, Yang Yu-Min, Wang Xiao-Dong
School of Biology & Basic Medical Sciences, Soochow University, Suzhou; Department of Histology and Embryology, Medical College, Nantong University, Nantong; Wuxi School of Medicine, Jiangnan University, Wuxi, Jiangsu Province, China.
Department of Histology and Embryology, Medical College, Nantong University, Nantong, Jiangsu Province, China.
Neural Regen Res. 2018 Jul;13(7):1231-1240. doi: 10.4103/1673-5374.235061.
The best tissue-engineered spinal cord grafts not only match the structural characteristics of the spinal cord but also allow the seed cells to grow and function in situ. Platelet-derived growth factor (PDGF) has been shown to promote the migration of bone marrow stromal cells; however, cytokines need to be released at a steady rate to maintain a stable concentration in vivo. Therefore, new methods are needed to maintain an optimal concentration of cytokines over an extended period of time to effectively promote seed cell localization, proliferation and differentiation. In the present study, a partition-type tubular scaffold matching the anatomical features of the thoracic 8-10 spinal cord of the rat was fabricated using chitosan and then subsequently loaded with chitosan-encapsulated PDGF-BB microspheres (PDGF-MSs). The PDGF-MS-containing scaffold was then examined in vitro for sustained-release capacity, biocompatibility, and its effect on neural progenitor cells differentiated in vitro from multilineage-differentiating stress-enduring cells (MUSE-NPCs). We found that pre-freezing for 2 hours at -20°C significantly increased the yield of partition-type tubular scaffolds, and 30 μL of 25% glutaraldehyde ensured optimal crosslinking of PDGF-MSs. The resulting PDGF-MSs cumulatively released 52% of the PDGF-BB at 4 weeks in vitro without burst release. The PDGF-MS-containing tubular scaffold showed suitable biocompatibility towards MUSE-NPCs and could promote the directional migration and growth of these cells. These findings indicate that the combination of a partition-type tubular scaffold, PDGF-MSs and MUSE-NPCs may be a promising model for the fabrication of tissue-engineered spinal cord grafts.
最佳的组织工程化脊髓移植物不仅要与脊髓的结构特征相匹配,还要使种子细胞能够在原位生长并发挥功能。血小板衍生生长因子(PDGF)已被证明可促进骨髓基质细胞的迁移;然而,细胞因子需要以稳定的速率释放,以在体内维持稳定的浓度。因此,需要新的方法来在较长时间内维持细胞因子的最佳浓度,以有效促进种子细胞的定位、增殖和分化。在本研究中,使用壳聚糖制备了一种与大鼠胸8 - 10脊髓解剖特征相匹配的分隔型管状支架,随后装载壳聚糖包封的PDGF - BB微球(PDGF - MSs)。然后在体外检测含PDGF - MSs的支架的缓释能力、生物相容性及其对从多谱系分化应激耐受细胞(MUSE - NPCs)体外分化而来的神经祖细胞的影响。我们发现,在-20°C预冻2小时可显著提高分隔型管状支架的产量,30 μL的25%戊二醛可确保PDGF - MSs的最佳交联。所得的PDGF - MSs在体外4周时累计释放了52%的PDGF - BB,无突发释放。含PDGF - MSs的管状支架对MUSE - NPCs表现出合适的生物相容性,并可促进这些细胞的定向迁移和生长。这些发现表明,分隔型管状支架、PDGF - MSs和MUSE - NPCs的组合可能是制备组织工程化脊髓移植物的一个有前景的模型。