Optimization of optical and mechanical properties of real architecture for 3-dimensional tissue equivalents: Towards treatment of limbal epithelial stem cell deficiency.
Optimization of optical and mechanical properties of real architecture for 3-dimensional tissue equivalents: Towards treatment of limbal epithelial stem cell deficiency.
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DOI：
10.1016/j.actbio.2015.06.007
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发表时间：
2015-09
影响因子：
9.7
通讯作者：
Daniels JT
中科院分区：
文献类型：
--
作者：
Massie I;Kureshi AK;Schrader S;Shortt AJ;Daniels JT
角膜缘上皮干细胞(LESC)缺乏可导致失明。将培养的人角膜缘上皮细胞(HLE)移植到人羊膜(HAM)上可以恢复视力，但临床上制作的移植物可能并不可靠。我们已经开发了一种可靠和强大的组织等效(TE)替代HAM，真正的三维组织架构(RAFT)。在此，我们旨在优化RAFT TE的光学和机械性能，用于治疗LESC缺乏症的临床应用。RAFT TE方案是可调的；不同的胶原蛋白浓度和体积会产生不同的RAFT TE。这些样本与取自胎盘盘近端和远端的火腿样本进行了比较。评估的结果是透明度、厚度、透光性、抗拉强度、易处理性、降解率和作为HLE培养底物的适宜性。近端的HAM样品比远端的HAM样品更厚、更坚固，光学性能更差。使用较高含量的胶原蛋白生产的木筏比使用较低含量的胶原蛋白生产的木筏更厚、更坚固，但光学性能较差。最理想的木筏TE很薄，透明，但仍然可以握手，使用0.6毫升3毫克/毫升的胶原蛋白制作而成。“最佳”木筏TE和火腿的降解率相似。HLE在“最佳”的RAFT TES上以与HAM相当的速度实现融合，并且细胞表达高水平的假定干细胞标记p63α。这些发现支持在肝移植治疗LESC缺乏症时使用RAFT TE。
Limbal epithelial stem cell (LESC) deficiency can cause blindness. Transplantation of cultured human limbal epithelial cells (hLE) on human amniotic membrane (HAM) can restore vision but clinical graft manufacture can be unreliable. We have developed a reliable and robust tissue equivalent (TE) alternative to HAM, Real Architecture for 3D Tissue (RAFT). Here, we aimed to optimize the optical and mechanical properties of RAFT TE for treatment of LESC deficiency in clinical application. The RAFT TE protocol is tunable; varying collagen concentration and volume produces differing RAFT TEs. These were compared with HAM samples taken from locations proximal and distal to the placental disc. Outcomes assessed were transparency, thickness, light transmission, tensile strength, ease of handling, degradation rates and suitability as substrate for hLE culture. Proximal HAM samples were thicker and stronger with poorer optical properties than distal HAM samples. RAFT TEs produced using higher amounts of collagen were thicker and stronger with poorer optical properties than those produced using lower amounts of collagen. The ‘optimal’ RAFT TE was thin, transparent but still handleable and was produced using 0.6 ml of 3 mg/ml collagen. Degradation rates of the ‘optimal’ RAFT TE and HAM were similar. hLE achieved confluency on ‘optimal’ RAFT TEs at comparable rates to HAM and cells expressed high levels of putative stem cell marker p63α. These findings support the use of RAFT TE for hLE transplantation towards treatment of LESC deficiency.