Seow Wei Yang, Kandasamy Karthikeyan, Peh Gary S L, Mehta Jodhbir S, Sun William
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #07-01, Singapore 138669.
Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
ACS Biomater Sci Eng. 2019 Aug 12;5(8):4067-4076. doi: 10.1021/acsbiomaterials.9b00610. Epub 2019 Jul 8.
We aimed to bioengineer a scaffold that can facilitate the transplantation of corneal endothelial cells (CEC), given the global shortage of cadaveric donor tissues. Although agarose (A) has outstanding biocompatibility and mechanical properties, it natively does not permit cell adhesion. In this study, agarose was modified with different attachment signals: GRGD (giving AR as product), lysine (AK), poly lysine (AP), and fish-derived gelatin (AG). Samples with varying conjugation ratios were prepared. All products formed bulk hydrogels, which were then collapsed into ultrathin membranes in a controlled environment. Membranes were evaluated for their ability to support attachment of various cell types. Cells, however, preferred the AG series of membrane. Notably, primary rabbit CEC remained attached and viable for ⩾4 weeks. The cells also stained positive for CD166, ZO-1 and Na/K ATPase, indicative of function. The hydrated AG membranes allowed >96% transmittance of visible light. The membranes were typically ∼15 μm thick and did not swell significantly after immersion in PBS. Tensile strength was 49-60 MPa, while young's modulus was 525-596 MPa. This membrane thus offers great promise as a scaffold for CEC during endothelial keratoplasty.
鉴于全球尸体供体组织短缺，我们旨在通过生物工程制造一种能够促进角膜内皮细胞（CEC）移植的支架。尽管琼脂糖（A）具有出色的生物相容性和机械性能，但它本身不允许细胞粘附。在本研究中，琼脂糖用不同的附着信号进行了修饰：GRGD（产物为AR）、赖氨酸（AK）、聚赖氨酸（AP）和鱼源明胶（AG）。制备了具有不同共轭比例的样品。所有产品都形成了块状水凝胶，然后在可控环境中塌陷成超薄膜。对薄膜支持各种细胞类型附着的能力进行了评估。然而，细胞更喜欢AG系列薄膜。值得注意的是，原代兔角膜内皮细胞在⩾4周内保持附着且存活。细胞对CD166、ZO - 1和钠钾ATP酶染色呈阳性，表明其功能正常。水合后的AG薄膜对可见光的透过率>96%。薄膜通常约15μm厚，浸入PBS后不会显著膨胀。拉伸强度为49 - 60MPa，而杨氏模量为525 - 596MPa。因此，这种薄膜作为内皮角膜移植术中角膜内皮细胞的支架具有很大的前景。
