Schmidt C, Fabinyi S, Rehfeldt S, Klöpzig S, Jentzen V, Bohrisch J, Messner A, Storsberg J
Fraunhofer-Institut für Angewandte Polymerforschung IAP, 14476, Potsdam-Golm, Deutschland.
HumanOptics AG, Erlangen, Deutschland.
Ophthalmologe. 2016 Oct;113(10):852-860. doi: 10.1007/s00347-016-0259-z.
Intrastromal insertion of Krumeich's corneal ring between graft and residual host corneal tissue appears to impair preripheral, superficial and superfluous vascularization of donor corneal tissue.
The purpose of this study was to investigate the cytotoxic effects of Krumeich's ring using tissue cultures composed of primary human dermal microvascular endothelial cells from adult donors (HMVEC).
Soluble growth medium extracts of the individual components of Krumeich's ring alloy were prepared and HMVEC were exposed to these extracts in triplicate for 1 day followed by investigation with 3‑(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Furthermore, HMVEC were cultured for 5 days on either Krumeich's ring or polypropylene (PP) discs coated with individual components of the Krumeich's ring alloy followed by double vital staining with fluorescein diacetate (FDA) and propidium iodide (PI).
The MTT assays revealed that higher doses of the extracts appeared to reduce the viability of HMVEC, while highly diluted extracts of molybdenum (Mo) powder appeared to increase the metabolic activity of HMVEC. The FDA-PI staining showed only a few live HMVEC on either cobalt (Co) or Mo-coated PP discs, compared to the respective titanium (Ti) and chromium (Cr) counterparts. Viable HMVEC appeared to attach to Krumeich's ring after a 5‑day incubation period.
The results confirm that Krumeich's ring does not exert measurable cytotoxic effects in our chosen assay system. High dilutions of medium-soluble Mo powder extracts appear to increase the metabolic activity of HMVEC.
在移植片与残留的宿主角膜组织之间基质内插入克鲁梅希角膜环似乎会损害供体角膜组织周边、浅层和多余的血管形成。
本研究的目的是使用来自成年供体的原代人真皮微血管内皮细胞（HMVEC）组成的组织培养物来研究克鲁梅希环的细胞毒性作用。
制备克鲁梅希环合金各成分的可溶性生长培养基提取物，将HMVEC一式三份暴露于这些提取物中1天，然后用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）试验进行检测。此外，将HMVEC在涂有克鲁梅希环合金各成分之一的克鲁梅希环或聚丙烯（PP）盘上培养5天，然后用荧光素二乙酸酯（FDA）和碘化丙啶（PI）进行双重活细胞染色。
MTT试验显示，较高剂量的提取物似乎会降低HMVEC的活力，而钼（Mo）粉的高度稀释提取物似乎会增加HMVEC的代谢活性。FDA-PI染色显示，与相应的钛（Ti）和铬（Cr）盘相比，钴（Co）或钼涂层PP盘上只有少数活的HMVEC。经过5天的孵育期后，有活力的HMVEC似乎附着在克鲁梅希环上。
结果证实，在我们选择的检测系统中，克鲁梅希环不会产生可测量的细胞毒性作用。中等可溶性钼粉提取物的高稀释度似乎会增加HMVEC的代谢活性。
