Pro-elastogenic effects of mesenchymal stem cell derived smooth muscle cells in a 3D collagenous milieu
Pro-elastogenic effects of mesenchymal stem cell derived smooth muscle cells in a 3D collagenous milieu
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DOI：
10.1016/j.actbio.2020.01.030
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发表时间：
2020-03-15
影响因子：
9.7
通讯作者：
Ramamurthi, Anand
中科院分区：
文献类型：
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作者：
Dahal, Shataakshi;Swaminathan, Ganesh;Ramamurthi, Anand
腹主动脉瘤（AAA）壁内的SMC对蛋白水解破坏的弹性基质结构的内在不良自动再生修复可防止这些壁扩张的生长停滞和消退。在先前的研究中，我们表明骨髓间充质干细胞衍生的平滑肌细胞（BM-SMCs）分泌的生物因子对非接触共培养的大鼠主动脉平滑肌细胞（EaRASMCs）具有显着的促弹性蛋白生成和抗蛋白水解作用，这支持了它们未来可能用于AAA的再生细胞治疗。我们还鉴定了一种稳定的BMSMC表型（cBM-SMC），其通过在存在PDGF（血小板衍生生长因子）和TGF-β 1（转化生长因子-β 1）的情况下在2D纤连蛋白基底上分化BM-MSC而产生，其表现出上级的弹性原性和促弹性原性/抗蛋白水解性质。在这项研究中，我们进一步研究了这些cBM-SMC在单独的3D胶原环境中以及与EaRASMC共培养中维持这些上级弹性蛋白生成特性的能力，以评估其作为AAA细胞治疗的替代细胞来源的潜力。我们的一些关键观察结果是在cBM-SMC的独立培养物以及cBM-SMC与EaRASMC 5的共培养物中的更高的收缩性和更大量的结构完整的弹性蛋白产生，如WG（Verhoeff-Van Gieson）染色和Pontamine天蓝色标记所示，以及在3D胶原环境中的独立培养物中的更低的MMP-9蛋白表达。我们的总体结果表明，cBM-SMC具有在3D胶原AAA环境中提供弹性生成动力的能力，否则这不利于弹性生成。因此，我们的研究强烈表明，cBM-SMCs作为一种潜在的细胞来源，用于细胞治疗，以增加弹性基质新组装和纤维形成，并减弱胶原环境中的蛋白水解，这是去弹性化的动脉壁的唤起。由主动脉壁中蛋白水解活性显著增加引起的相关死亡。逆转这种情况的病理生理学是具有挑战性的，因为主动脉平滑肌细胞的弹性蛋白再生本质上很差。AM的当前管理仅限于被动监测疾病，直到它变得足够大以接受手术干预，并且目前不存在基于药物的治疗。在这种情况下，基于细胞的治疗可以是潜在的替代治疗，因为它为平滑肌细胞提供弹性动力，补偿死亡的平滑肌细胞，并作为弹性蛋白的强大来源，同时以最小的侵入性递送。因此，这项工作将在组织工程和再生医学领域产生重大影响。(C)2020由Elsevier Ltd代表Acta Materialia Inc.发布。
Intrinsically poor auto-regenerative repair of proteolytically-disrupted elastic matrix structures by resident SMCs in the wall of abdominal aortic aneurysms (AAAs) prevents growth arrest and regression of these wall expansions. Supporting their possible future use in a regenerative cell therapy for AAAs, in a prior study, we showed that bone marrow mesenchymal stem cell-derived Smooth Muscle Cells (BM-SMCs) secrete biological factors that have significant pro-elastogenic and anti-proteolytic effects on aneurysmal rat aortic SMCs (EaRASMCs) in non-contact co-cultures. We also identified one stable BMSMC phenotype (cBM-SMC) generated by differentiating BM-MSCs on a 2D fibronectin substrate in the presence of PDGF (Platelet Derived Growth Factor) and TGF-beta 1 (Transforming Growth Factor-beta 1) that exhibited superior elastogenicity and pro-elastogenic/anti-proteolytic properties. In this study, we further investigated the ability of these cBM-SMCs to maintain these superior elastogenic properties in a 3D collagenous milieu alone and in co-culture with EaRASMC to evaluate their potential as an alternative cell source for cell therapy in AAA. Some of our key observations were higher contractility and greater amount of structurally intact elastin production in both standalone culture of cBM-SMCs as well as co culture of cBM-SMCs with EaRASMC5 as shown by WG (Verhoeff-Van Gieson) staining and Pontamine Sky Blue labeling and lower MMP-9 protein expression in standalone culture in 3D collagenous environment. Our overall result indicates that cBM-SMCs possess the ability to provide elastogenic impetus in a 3D collagenous AAA milieu which is otherwise not conducive to elastogenesis. Therefore our study strongly suggest the utility of cBM-SMCs as a potential cell source for cell therapy to augment elastic matrix neo-assembly and fiber formation and attenuate proteolysis in a collagenous milieu that is evocative of the de-elasticized aneurysmal wall.Statement of SignificanceAbdominal aortic aneurysm (AM) or ballooning of the aorta is one of the leading causes of cardiovascular disease (CVD) related death caused by significantly increased proteolytic activity in the aortic wall. Reversing pathophysiology of this condition is challenging due to intrinsically poor regeneration of elastin by aortic smooth muscle cells. Current management of AM is limited to passive monitoring of the disease until it becomes large enough to receive surgical intervention and no drug based therapy currently exists. Cell based therapy can be a potential alternative treatment in this scenario because it provides elastogenic impetus to the aneurysmal SMCs, compensates for the dead SMCs and serves as a robust source of elastin while being delivered with minimal invasiveness. Hence this work will have significant impact in the field of tissue engineering and regenerative medicine. (C) 2020 Published by Elsevier Ltd on behalf of Acta Materialia Inc.