Faggioli Marika, Moro Arianna, Butt Salman, Todesco Martina, Sandrin Deborah, Borile Giulia, Bagno Andrea, Fabozzo Assunta, Romanato Filippo, Marchesan Massimo, Imran Saima, Gerosa Gino
Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padua, I-35128 Padua, Italy.
L.I.F.E.L.A.B. Program, Consorzio per la Ricerca Sanitaria (CORIS), Veneto Region, I-35127 Padua, Italy.
Polymers (Basel). 2022 Mar 17;14(6):1226. doi: 10.3390/polym14061226.
The most common aortic valve diseases in adults are stenosis due to calcification and regurgitation. In pediatric patients, aortic pathologies are less common. When a native valve is surgically replaced by a prosthetic one, it is necessary to consider that the latter has a limited durability. In particular, current bioprosthetic valves have to be replaced after approximately 10 years; mechanical prostheses are more durable but require the administration of permanent anticoagulant therapy. With regard to pediatric patients, both mechanical and biological prosthetic valves have to be replaced due to their inability to follow patients' growth. An alternative surgical substitute can be represented by the acellular porcine aortic valve that exhibits less immunogenic risk and a longer lifespan. In the present study, an efficient protocol for the removal of cells by using detergents, enzyme inhibitors, and hyper- and hypotonic shocks is reported. A new detergent (Tergitol) was applied to replace TX-100 with the aim to reduce toxicity and maximize ECM preservation. The structural integrity and efficient removal of cells and nuclear components were assessed by means of histology, immunofluorescence, and protein quantification; biomechanical properties were also checked by tensile tests. After decellularization, the acellular scaffold was sterilized with a standard protocol and repopulated with bone marrow mesenchymal stem cells to analyze its biocompatibility profile.
成人中最常见的主动脉瓣疾病是钙化导致的狭窄和反流。在儿科患者中，主动脉病变较少见。当天然瓣膜通过人工瓣膜进行手术置换时，有必要考虑到后者的耐久性有限。特别是，目前的生物人工瓣膜大约10年后就必须更换；机械瓣膜更耐用，但需要长期进行抗凝治疗。对于儿科患者，由于机械和生物人工瓣膜都无法适应患者的生长，因此都必须更换。一种替代的手术替代品可以是无细胞猪主动脉瓣，其免疫原性风险较低，使用寿命更长。在本研究中，报告了一种使用去污剂、酶抑制剂以及高渗和低渗休克去除细胞的有效方案。应用一种新型去污剂（Tergitol）来替代TX-100，以降低毒性并最大限度地保留细胞外基质。通过组织学、免疫荧光和蛋白质定量评估细胞和核成分的结构完整性及有效去除情况；还通过拉伸试验检查生物力学性能。脱细胞后，用标准方案对无细胞支架进行消毒，并用骨髓间充质干细胞重新填充，以分析其生物相容性。
