Eye Center, Medical Center, Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, Germany.
Department of Ophthalmology, University Hospital Erlangen, Friedrich-Alexander-University of Erlangen-Nürnberg, Schwabachanlage 6, D-91054 Erlangen, Germany.
Int J Mol Sci. 2021 Sep 17;22(18):10067. doi: 10.3390/ijms221810067.
The transplantation of ex vivo expanded limbal epithelial progenitor cells (LEPCs) on amniotic membrane or fibrin gel is an established therapeutic strategy to regenerate the damaged corneal surface in patients with limbal stem cell deficiency (LSCD), but the long-term success rate is restricted. A scaffold with niche-specific structure and extracellular matrix (ECM) composition might have the advantage to improve long-term clinical outcomes, in particular for patients with severe damage or complete loss of the limbal niche tissue structure. Therefore, we evaluated the decellularized human limbus (DHL) as a biomimetic scaffold for the transplantation of LEPCs. Corneoscleral tissue was decellularized by sodium deoxycholate and deoxyribonuclease I in the presence or absence of dextran. We evaluated the efficiency of decellularization and its effects on the ultrastructure and ECM composition of the human corneal limbus. The recellularization of these scaffolds was studied by plating cultured LEPCs and limbal melanocytes (LMs) or by allowing cells to migrate from the host tissue following a lamellar transplantation ex vivo. Our decellularization protocol rapidly and effectively removed cellular and nuclear material while preserving the native ECM composition. In vitro recellularization by LEPCs and LMs demonstrated the good biocompatibility of the DHL and intrastromal invasion of LEPCs. Ex vivo transplantation of DHL revealed complete epithelialization as well as melanocytic and stromal repopulation from the host tissue. Thus, the generated DHL scaffold could be a promising biological material as a carrier for the transplantation of LEPCs to treat LSCD.
体外扩增的角膜缘上皮祖细胞（LEPCs）在羊膜或纤维蛋白凝胶上的移植是一种已建立的治疗策略，可用于再生患有角膜缘干细胞缺乏症（LSCD）的患者受损的角膜表面，但长期成功率受到限制。具有特定龛位结构和细胞外基质（ECM）组成的支架可能具有改善长期临床结果的优势，特别是对于具有严重损伤或完全丧失角膜缘龛位组织结构的患者。因此，我们评估了去细胞人角膜缘（DHL）作为 LEPCs 移植的仿生支架。通过在有或没有葡聚糖的情况下用脱氧胆酸钠和脱氧核糖核酸酶 I 对角膜缘组织进行去细胞化处理。我们评估了去细胞化的效率及其对人眼角膜缘的超微结构和 ECM 组成的影响。通过培养 LEPCs 和角膜缘黑素细胞（LMs）在这些支架上进行再细胞化，或允许细胞从宿主组织中迁移，进行体外层状移植。我们的去细胞化方案快速有效地去除了细胞和核材料，同时保留了天然的 ECM 组成。LEPCs 和 LMs 的体外再细胞化证明了 DHL 的良好生物相容性和 LEPCs 的基质内浸润。DHL 的体外移植显示出完全的上皮化以及来自宿主组织的黑素细胞和基质再填充。因此，所产生的 DHL 支架可以作为 LEPCs 移植治疗 LSCD 的载体，是一种很有前途的生物材料。
