Recellularization of decellularized allograft scaffolds in ovine great vessel reconstructions
Recellularization of decellularized allograft scaffolds in ovine great vessel reconstructions
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DOI：
10.1016/j.athoracsur.2004.09.033
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发表时间：
2005-03-01
影响因子：
4.6
通讯作者：
Hopkins, R
中科院分区：
文献类型：
--
作者：
Ketchedjian, A;Jones, AL;Hopkins, R
背景脱细胞同种异体移植物组织已被确定为潜在的细胞外基质（EM支架），再细胞化组织工程血管和瓣膜替代品的基础。降低的抗原性和再细胞化的能力表明这样的构建体可能具有有利的耐久性。去污剂/酶脱细胞方法去除细胞和细胞碎片，同时留下完整的结构蛋白“支架”。在一个长期植入模型中测试了用阴离子去污剂/酶方法脱细胞的同种异体肺动脉组织，该模型使用幼年绵羊大血管中的动脉壁修复。对于三种不同的支架方案中的每一种，植入12只试验绵羊（n = 4），所述方案比较了传统的二甲基亚砜冷冻保存、冷冻保存后去细胞化和新鲜组织的去细胞化。另外4只绵羊作为对照，n = 2假手术，n = 2新鲜组织）。将贴片成型并植入肺动脉中，并随时间（10至20周）进行测量。外植体组织病理学确定再细胞化形态以及钙，胶原蛋白和弹性蛋白在移植组织中的分布。与传统冷冻保存的组织不同，脱细胞组织在植入前不含残留细胞或细胞碎片，这与PRA的可测量减少相关。脱细胞外植体表现出受体细胞通过基质的时间依赖性迁移，典型的a-平滑肌肌动蛋白染色阳性，无钙化。所展示的特性似乎与可植入组织工程支架所需的特性一致。所描述的去细胞化方法似乎产生了生物学上合适的ECM支架，用于表型上合适的细胞的体内迁移，同时避免了抗原性和钙化。
Background. Decellularized allograft tissues have been identified as a potential extracellular matrix (EM scaffold on which to base recellularized tissue-engineered vascular and valvular substitutes. Decreased antigenicity and the capacity to recellularize suggest that such constructs may have favorable durability. Detergent/enzyme decellularization methods remove cells and cellular debris while leaving intact structural protein "scaffolds." Allograft pulmonary artery tissues decellularized with an anionic detergent/enzyme methodology were tested in a long-term implantation model that used arterial wall repairs in the great vessels of juvenile sheep.Methods. Twelve test sheep were implanted (n = 4) for each of three different scaffold protocols that compared traditional dimethylsulfoxide cryopreservation, cryopreservation followed by decellularization, and decellularization of fresh tissue. Four additional sheep served as controls In = 2 sham, n = 2 fresh tissue). Patches were fashioned and implanted into pulmonary artery and measured over time (10 to 20 weeks). Explant histopathology determined recellularization morphology as well as calcium, collagen, and elastin distribution within explanted tissue.Results. Unlike traditionally cryopreserved tissues, the decellularized tissues contained no residual cells or cellular debris before implantation, which correlated with measurable reductions in PRA. Decellularized explants demonstrated time-dependent migration of recipient cells through matrix, typically staining positive for a-smooth muscle actin with no calcification.Conclusions. The properties demonstrated seem consistent with characteristics necessary for implantable tissue-engineered scaffolds. The decellularization method described appears to create a biologically suitable ECM scaffold for in vivo migration of phenotypically appropriate cells while avoiding antigenicity and calcification.