Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity.
Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity.
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DOI：
10.3389/fbioe.2022.883977
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发表时间：
2022
影响因子：
5.7
通讯作者：
Griffith, May
中科院分区：
文献类型：
--
作者：
Simpson, Fiona C.;Islam, Mohammed Mirazul;Buznyk, Oleksiy;Edin, Elle;Groleau, Marc;Kozak-Ljunggren, Monika;Magrelli, Federica M.;AbuSamra, Dina B.;Argueeso, Pablo;Chodosh, James;Liszka, Aneta;Fagerholm, Per;Griffith, May
可生物降解的胶原基植入物的灭菌是具有挑战性的，因为辐照灭菌方法可以改变其机械性能。电子束（EB）辐照是一种终端灭菌方法，已用于生物衍生植入物。在这里，重组人iii型胶原-磷酸胆碱（RHCIII-MPC）水凝胶以17,19或21 kGy的EB剂量照射，并评估其随后的生物相容性和促进兔角膜再生的能力。未辐照的水凝胶保存在1%氯仿磷酸盐缓冲盐水（C-PBS）中作为对照。辐照样品和未辐照样品在光学或物理性质（抗拉强度、模量、弹性）或支持细胞生长的能力方面没有显著差异。然而，与未照射的对照组相比，照射后的植入物对高水平的胶原酶更敏感，与EB和对照组相比，C-PBS植入物在72小时内的细胞生长增加。以17千吉/小时的速度进行电子照射或电子照射后冷冻（EB- f）以延长保质期的角膜植入物没有显示出辐照的不良生物效应。EB、EB- f和C-PBS植入的角膜都能迅速重新上皮化，但表现出轻微的新生血管，在6个月后消退。术后6个月再生的新角膜透明。体内共聚焦显微镜证实上皮、间质、亚基底神经和未手术内皮形态正常。组织学结果显示，所有再生角膜形态与正常角膜相似。免疫组织化学显示存在分化的角膜上皮和功能性泪膜。综上所述，电子束角膜植入物的表现与非辐照对照植入物一样，产生了完全再生的新角膜，有新的神经，没有血管或可能妨碍视力或角膜功能的炎症。因此，下一步有必要在临床应用前进行一项完整的验证研究，以确定EB照射作为角膜种植体消毒的有效手段。
Sterilization of biodegradable, collagen-based implants is challenging as irradiation sterilization methods can alter their mechanical properties. Electron beam (EB) irradiation is a terminal sterilization method that has been used for biologically-derived implants. Here, recombinant human collagen type III-phosphorylcholine (RHCIII-MPC) hydrogels were irradiated with EB doses of 17, 19, or 21 kGy and their subsequent biocompatibility and ability to promote regeneration in rabbit corneas was evaluated. Unirradiated hydrogels stored in 1% chloroform in phosphate-buffered saline (C-PBS) were the controls. There were no significant differences between irradiated and non-irradiated samples in optical or physical properties (tensile strength, modulus, elasticity), or the ability to support cell growth. However, irradiated implants were more sensitive to high levels of collagenase than unirradiated controls and the C-PBS implants had increased cell growth compared to EB and controls at 72 h. Corneal implants e-beamed at 17 kGy or e-beamed and subsequently frozen (EB-F) to increase shelf-life showed no adverse biological effects of the irradiation. EB, EB-F, and C-PBS implanted corneas all rapidly re-epithelialized but showed mild neovascularization that resolved over 6 months. The regenerated neo-corneas were transparent at 6 months post-operation. In vivo confocal microscopy confirmed normal morphology for the epithelium, stroma, sub-basal nerves and unoperated endothelium. Histology showed that all the regenerated corneas were morphologically similar to the normal. Immunohistochemistry indicated the presence of a differentiated corneal epithelium and functional tear film. In conclusion, the e-beamed corneal implants performed as well as non-irradiated control implants, resulting in fully regenerated neo-corneas with new nerves and without blood vessels or inflammation that may impede vision or corneal function. Therefore, a complete validation study to establish EB irradiation as an effective means for corneal implant sterilization prior to clinical application is necessary as a next step.
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影响因子：
14
作者：
Fagerholm, Per;Lagali, Neil S.;Griffith, May
通讯作者：
Griffith, May
影响因子：
4.4
作者：
Hackett, Joanne M.;Lagali, Neil;Fagerholm, Per
通讯作者：
Fagerholm, Per
影响因子：
4.8
作者：
Hoburg, Arnd T.;Keshlaf, Salahedeen;Scheffler, Sven
通讯作者：
Scheffler, Sven
影响因子：
8.2
作者：
Dai Z;Ronholm J;Tian Y;Sethi B;Cao X
通讯作者：
Cao X
影响因子：
9.7
作者：
Islam, M. Mirazul;Cepla, Vytautas;Griffith, May
通讯作者：
Griffith, May