Biological Sciences, Faculty of Medicine, Dentistry and Life Sciences, University of Chester, Parkgate Road, Chester, CH1 4BJ, United Kingdom.
Centre for Experimental Medicine, Queen's University, 97 Lisburn Road, Belfast, BT9 7BL, Northern Ireland, UK.
Biochimie. 2018 Dec;155:26-36. doi: 10.1016/j.biochi.2018.04.011. Epub 2018 Apr 19.
The multifactorial complexity of spinal cord injuries includes the formation of a glial scar, of which chondroitin sulphated proteoglycans (CSPG) are an integral component. Previous studies have shown CSPG to have inhibitory effects on endothelial and neuronal cell growth, highlighting the difficulty of spinal cord regeneration. Mesenchymal stem/stromal cells (MSC) are widely used as a cell therapy, and there is mounting evidence for their angiogenic and neurotrophic paracrine properties. However, in vivo studies have observed poor engraftment and survival of MSC when injected into SCI. Currently, it is not known whether increasing CSPG concentrations seen after SCI may affect MSC; therefore we have investigated the effects of CSPG exposure to MSC in vitro. CSPG-mediated inhibition of MSC adhesion was observed when MSC were cultured on substrates of increasing CSPG concentration, however MSC viability was not affected even up to five days of culture. Culture conditioned medium harvested from these cultures (primed MSC CM) was used as both culture substrata and soluble medium for EA.hy926 endothelial cells and SH-SY5Y neuronal cells. MSC CM was angiogenic, promoting endothelial cell adhesion, proliferation and tubule formation. However, exposing MSC to CSPG reduced the effects of CSPG-primed MSC CM on endothelial cell adhesion and proliferation, but did not reduce MSC-induced endothelial tubule formation. Primed MSC CM also promoted neuronal cell adhesion, which was reduced following exposure to CSPG. There were no marked differences in neurite outgrowth in MSC CM from CSPG primed MSC cultures versus control conditions, although non-primed MSC CM from the same donors was found to significantly enhance neurite outgrowth. Taken together, these studies demonstrate that MSC are resilient to CSPG exposure, but that there is a marked effect of CSPG on their paracrine regenerative activity. The findings increase our understanding of how the wound microenvironment after SCI can mitigate the beneficial effects of MSC transplantation.
脊髓损伤的多因素复杂性包括胶质瘢痕的形成，其中软骨素硫酸盐蛋白聚糖（CSPG）是其组成部分。先前的研究表明 CSPG 对内皮细胞和神经元细胞的生长具有抑制作用，这突出了脊髓再生的困难。间充质干细胞（MSC）被广泛用作细胞治疗，并且越来越多的证据表明其具有血管生成和神经营养的旁分泌特性。然而，在注射到脊髓损伤中时，体内研究观察到 MSC 的植入和存活不良。目前，尚不清楚脊髓损伤后 CSPG 浓度的增加是否会影响 MSC；因此，我们已经研究了 CSPG 暴露对 MSC 的体外影响。当 MSC 在 CSPG 浓度逐渐增加的基质上培养时，观察到 CSPG 介导的 MSC 黏附抑制，但即使在培养五天后，MSC 的活力也没有受到影响。从这些培养物中收获的培养条件培养基（预刺激 MSC CM）既用作培养底物，也用作 EA.hy926 内皮细胞和 SH-SY5Y 神经元细胞的可溶性培养基。MSC CM 具有血管生成作用，可促进内皮细胞黏附、增殖和管腔形成。然而，将 MSC 暴露于 CSPG 会降低 CSPG 预刺激 MSC CM 对内皮细胞黏附和增殖的作用，但不会降低 MSC 诱导的内皮管腔形成。预刺激 MSC CM 还促进神经元细胞黏附，而暴露于 CSPG 后则减少了这种黏附。在 CSPG 预刺激 MSC 培养物的 MSC CM 中，神经突生长没有明显差异，而来自同一供体的非预刺激 MSC CM 被发现可显著增强神经突生长。总之，这些研究表明，MSC 对 CSPG 暴露具有弹性，但 CSPG 对其旁分泌再生活性有明显影响。这些发现增加了我们对脊髓损伤后伤口微环境如何减轻 MSC 移植有益作用的理解。
