Shear and endothelial induced late-stage calcific aortic valve disease-on-a-chip develops calcium phosphate mineralizations
Shear and endothelial induced late-stage calcific aortic valve disease-on-a-chip develops calcium phosphate mineralizations
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DOI：
10.1039/d1lc00931a
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发表时间：
2022-02-21
期刊：
影响因子：
6.1
通讯作者：
Mahler, Gretchen J.
中科院分区：
文献类型：
--
作者：
Mendoza, Melissa;Chen, Mei-Hsiu;Mahler, Gretchen J.
钙化性主动脉瓣病(CAVD)是导致严重主动脉狭窄的一种活跃的病理生物学过程，唯一的治疗方法是瓣膜置换术。晚期CAVD的特征是纤维组织中钙化、胶原解体和糖胺聚糖沉积，如硫酸软骨素(CS)。我们开发了一种三维微流控装置，用于研究切应力(1或20达因/厘米(2))、CS(1或20 mg mL(-1))和内皮细胞存在对钙化的影响。CAVD芯片由I型胶原水凝胶组成，猪主动脉瓣间质细胞植入其中，猪主动脉瓣内皮细胞种植在基质上，最长可达21d。在这里，我们展示了这种芯片上的CAVD是第一个形成类似人类的钙化结节，其钙磷矿化成熟度不同，是由高剪切力和内皮细胞，特别是二钙和八钙磷酸钙引起的。长期的共培养微流控研究证实了细胞存活和21天内磷酸钙的形成。鉴于CAVD没有针对性的治疗方法，建立一个与生理相关的主动脉瓣试验台可能会导致临床前研究的进展。
Calcific aortic valve disease (CAVD) is an active pathobiological process leading to severe aortic stenosis, where the only treatment is valve replacement. Late-stage CAVD is characterized by calcification, disorganization of collagen, and deposition of glycosaminoglycans, such as chondroitin sulfate (CS), in the fibrosa. We developed a three-dimensional microfluidic device of the aortic valve fibrosa to study the effects of shear stress (1 or 20 dyne per cm(2)), CS (1 or 20 mg mL(-1)), and endothelial cell presence on calcification. CAVD chips consisted of a collagen I hydrogel, where porcine aortic valve interstitial cells were embedded within and porcine aortic valve endothelial cells were seeded on top of the matrix for up to 21 days. Here, we show that this CAVD-on-a-chip is the first to develop human-like calcified nodules varying in calcium phosphate mineralization maturity resulting from high shear and endothelial cells, specifically di- and octa-calcium phosphates. Long-term co-culture microfluidic studies confirmed cell viability and calcium phosphate formations throughout 21 days. Given that CAVD has no targeted therapies, the creation of a physiologically relevant test-bed of the aortic valve could lead to advances in preclinical studies.