Preparation, preservation, and morphological evaluation of the donor graft for descemet membrane endothelial keratoplasty: an experimental study
Preparation, preservation, and morphological evaluation of the donor graft for descemet membrane endothelial keratoplasty: an experimental study
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后弹力层内皮角膜移植供体的制备、保存及形态学评价：实验研究
DOI：
10.3760/cma.j.issn.0366-6999.20133097
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发表时间：
2014-05
影响因子：
6.1
通讯作者：
Hong Jing
中科院分区：
文献类型：
--
作者：
Sun Yiqian;Peng Rongmei;Hong Jing
背景：目前已有多种方法获取和保存Descemet膜(DM)和完整的内皮细胞，但关于Descemet膜内皮细胞移植(Descemet Memet Memet Endothalkeratportion，DMEK)移植准备后内皮细胞的形态评价尚无文献报道。本研究的目的是建立和改进一种简便的制备、保存和评价DMEK供体移植物的方法。方法用29G针将1ml无菌空气注入Schwalbe线外的小边缘，形成气泡，以获得供体移植物。另一根针通过基质插入气泡，以吸入空气或用器官培养液取代一半空气。采用台盼蓝标记小切口的位置，提高手术成功率。冰冻切片苏木精-伊红(HE)染色。根据气泡大小分为4组：A组(正常对照组)，不加任何手术方法；B组，零压气泡；C组，全压气泡；D组，半压气泡。将4组移植物保存24小时，观察气泡对细胞的影响。用茜素红和扫描电子显微镜对样品的大体和超微结构进行了表征。结果供体移植物通过气泡法获得，并通过台盼蓝染色获得。HE染色切片显示为无间质的纯移植物。光镜下未见明显改变。A组：扫描电镜下可见多角形内皮细胞融合层，微绒毛呈散在分布，呈交错连接特征。B组细胞间边界变薄。C组交错多为扁平，微绒毛较少。D组除微绒毛减少外，其余变化不大。结论供体移植物制备方法简便、有效。适当降低气压可以保护和保存血管内皮细胞。
Background Though there have been various methods for harvesting and preserving descemet membrane (DM) and intact endothelium, there is no literature about the morphological evaluation of endothelium after graft preparation for descemet membrane endothelial keratoplasty (DMEK). The aim of this study was to establish and improve a simple method for preparing, preserving, and morphologically evaluating the donor graft for DMEK. Methods To obtain a donor graft, an air bubble was formed by injecting a 29 G needle with 1 ml sterile air into a small edge created outside the Schwalbe line. Another needle was inserted into the bubble through the stroma to aspirate the air or replace half the air with organ culture medium. Trypan blue was used to mark the location for small incision to improve the success rate. Frozen sections were stained with hematoxylin and eosin (HE). Based on the air bubble, DM grafts were divided into four groups: group A (normal control), graft without any operative technique; group B, graft with zero‐pressure air bubble; group C, graft with full‐pressure air bubble; group D, graft with half‐pressure air bubble. The four groups of grafts were preserved for 24 hours to observe the effect of bubbles on cells. The gross and ultrastructure morphologies were evaluated using alizarin red and scanning electron microscopy (SEM), respectively. Results Donor grafts were harvested via the air bubble technique, facilitated by prior trypan blue staining. HE‐stained sections revealed a pure graft without stroma. There were no significant changes under light microscope. In group A, SEM revealed a confluent layer of polygonal endothelium with distributed microvilli exhibiting characteristics of interdigitating junctions. In group B, intercellular borders became thinner. In group C, interdigitations were almost flat and microvilli were observed less frequently. In group D, other than less microvilli, there were minimal changes. Conclusions The donor graft preparation method appears to be effective and convenient. Properly decreasing the air pressure could protect and preserve the endothelium.
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影响因子：
2.8
作者：
S. Chang;J. Jun;K. Hong
通讯作者：
S. Chang;J. Jun;K. Hong
影响因子：
2.8
作者：
D. Venzano;P. Pagani;Nadia Randazzo;F. Cabiddu;C. Traverso
通讯作者：
D. Venzano;P. Pagani;Nadia Randazzo;F. Cabiddu;C. Traverso
影响因子：
4.2
作者：
Melles, Gerrit R. J.;Ong, T. San;Van der Wees, Jacqueline
通讯作者：
Van der Wees, Jacqueline
影响因子：
4.2
作者：
Teresa S. Ignacio;Thao T B Nguyen;M. Sarayba;P. Sweet;O. Piovanetti;R. Chuck;A. Behrens
通讯作者：
Teresa S. Ignacio;Thao T B Nguyen;M. Sarayba;P. Sweet;O. Piovanetti;R. Chuck;A. Behrens
影响因子：
2.8
作者：
Lie, Jessica T.;Birbal, Renuka;Melles, Gerrit R. J.
通讯作者：
Melles, Gerrit R. J.