The feasibility and efficacy of preparing porcine Descemet’s membrane endothelial keratoplasty (DMEK) grafts by two techniques: An ex‐vivo investigation for future xeno‐DMEK

Background Descemet’s membrane endothelial keratoplasty (DMEK) might be a promising technique for future xeno‐corneal transplantation due to its ultrathin graft, extremely low rejection occurrence, suture‐free graft fixation, and minimal immunosuppressive regime usage. The aim of this study is to ex...

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Veröffentlicht in:Xenotransplantation (Københaven) 2018-09, Vol.25 (5), p.e12407-n/a
Hauptverfasser: Liu, Yang, Zhang, Jing, Zhang, Yingnan, Yin, Mingyang, Miao, Sen, Liang, Qingfeng, Pan, Zhiqiang
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container_title Xenotransplantation (Københaven)
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creator Liu, Yang
Zhang, Jing
Zhang, Yingnan
Yin, Mingyang
Miao, Sen
Liang, Qingfeng
Pan, Zhiqiang
description Background Descemet’s membrane endothelial keratoplasty (DMEK) might be a promising technique for future xeno‐corneal transplantation due to its ultrathin graft, extremely low rejection occurrence, suture‐free graft fixation, and minimal immunosuppressive regime usage. The aim of this study is to explore the feasibility and efficacy of preparing porcine DMEK grafts by 2 techniques and investigate the graft ultrastructure. Methods Two mainstream techniques, mechanical stripping technique and liquid bubble technique, were modified to prepare the porcine DMEK grafts. In all, 40 corneas harvested from WZS‐pigs (aged 10‐12 months) were subjected to the techniques (20 corneas for each technique). The success rate, time consumption, and endothelial cell density (ECD) before and after preparation were recorded and compared between the 2 techniques. And the ultrastructure of the porcine DEMK graft was investigated by transmission electron microscope. In addition, 9 WZS‐pigs with different ages were sacrificed to explore the correlation between the thickness of Descemet’s membrane and porcine age. Results After modifying several technical details, the porcine DMEK grafts were successfully prepared by either mechanical stripping technique or liquid bubble technique, and the mark technique to distinguish the 2 sides of the graft was also explored. In all, 13 DMEK grafts (65%) were prepared successfully by the mechanical stripping technique, whereas 14 successful cases (70%) were prepared by the liquid bubble technique. The success rates between the 2 techniques showed no significant difference (P = .847). However, the mechanical stripping technique was significantly time‐consuming when compared with the liquid bubble technique (P 
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The aim of this study is to explore the feasibility and efficacy of preparing porcine DMEK grafts by 2 techniques and investigate the graft ultrastructure. Methods Two mainstream techniques, mechanical stripping technique and liquid bubble technique, were modified to prepare the porcine DMEK grafts. In all, 40 corneas harvested from WZS‐pigs (aged 10‐12 months) were subjected to the techniques (20 corneas for each technique). The success rate, time consumption, and endothelial cell density (ECD) before and after preparation were recorded and compared between the 2 techniques. And the ultrastructure of the porcine DEMK graft was investigated by transmission electron microscope. In addition, 9 WZS‐pigs with different ages were sacrificed to explore the correlation between the thickness of Descemet’s membrane and porcine age. Results After modifying several technical details, the porcine DMEK grafts were successfully prepared by either mechanical stripping technique or liquid bubble technique, and the mark technique to distinguish the 2 sides of the graft was also explored. In all, 13 DMEK grafts (65%) were prepared successfully by the mechanical stripping technique, whereas 14 successful cases (70%) were prepared by the liquid bubble technique. The success rates between the 2 techniques showed no significant difference (P = .847). However, the mechanical stripping technique was significantly time‐consuming when compared with the liquid bubble technique (P &lt; .0001). The ECDs reduced significantly after preparation no matter what techniques were used (P &lt; .0001), but the ECD after the liquid bubble preparation was significantly higher than the ECD after mechanical stripping (P = .032). The ECD reduction positively correlated to the time consumption for both mechanical stripping technique (P = .0014, R2 = 0.621) and liquid bubble technique (P = .013, R2 = 0.412). The ultrastructure showed the graft was comprised of stromal residuals, non‐banded layer, and endothelial layer. Unlike human Descemet’s membrane (DM), anterior banded layer was not observed. The thickness of porcine DM increased with the age, and a significant positive correlation between them was found (P &lt; .0001, R2 = 0.949), and the predict equation was Y = 0.3764*X + 7.378 (Y indicates the thickness, whereas X indicates the age). Conclusions Porcine DMEK grafts could be prepared either by mechanical stripping technique or liquid bubble technique, and the liquid bubble technique seems superior over the mechanical stripping technique regarding time consumption and ECD preservation. Although there are several technical barriers to overcome, xeno‐DMEK might be a promising direction for future xeno‐corneal transplantation.</description><identifier>ISSN: 0908-665X</identifier><identifier>EISSN: 1399-3089</identifier><identifier>DOI: 10.1111/xen.12407</identifier><identifier>PMID: 29756410</identifier><language>eng</language><publisher>Denmark: Wiley Subscription Services, Inc</publisher><subject>Adult ; Age ; Aged ; Animals ; Cell density ; Cornea ; Corneal Diseases - immunology ; Corneal Diseases - surgery ; Corneal transplantation ; Corneal Transplantation - methods ; Descemet Membrane - immunology ; Descemet’s membrane endothelial keratoplasty ; Endothelial cells ; ex‐vivo ; Female ; graft preparation ; Graft rejection ; Graft Survival - immunology ; Humans ; Immunosuppression ; Investigations ; liquid bubble technique ; mechanical stripping technique ; Middle Aged ; porcine cornea ; Preservation ; Swine ; Tissue and Organ Harvesting ; Tissue Donors ; Transplantation, Heterologous - methods ; Transplants - immunology ; Ultrastructure ; Xenografts ; xeno‐corneal transplantation</subject><ispartof>Xenotransplantation (Københaven), 2018-09, Vol.25 (5), p.e12407-n/a</ispartof><rights>2018 John Wiley &amp; Sons A/S. Published by John Wiley &amp; Sons Ltd.</rights><rights>2018 John Wiley &amp; Sons A/S</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3537-e1d43f1801026045844c3cbe3a8356576bb382cbe8fbb9e82765daa36df94ded3</citedby><cites>FETCH-LOGICAL-c3537-e1d43f1801026045844c3cbe3a8356576bb382cbe8fbb9e82765daa36df94ded3</cites><orcidid>0000-0002-6854-0615</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fxen.12407$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fxen.12407$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29756410$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liu, Yang</creatorcontrib><creatorcontrib>Zhang, Jing</creatorcontrib><creatorcontrib>Zhang, Yingnan</creatorcontrib><creatorcontrib>Yin, Mingyang</creatorcontrib><creatorcontrib>Miao, Sen</creatorcontrib><creatorcontrib>Liang, Qingfeng</creatorcontrib><creatorcontrib>Pan, Zhiqiang</creatorcontrib><title>The feasibility and efficacy of preparing porcine Descemet’s membrane endothelial keratoplasty (DMEK) grafts by two techniques: An ex‐vivo investigation for future xeno‐DMEK</title><title>Xenotransplantation (Københaven)</title><addtitle>Xenotransplantation</addtitle><description>Background Descemet’s membrane endothelial keratoplasty (DMEK) might be a promising technique for future xeno‐corneal transplantation due to its ultrathin graft, extremely low rejection occurrence, suture‐free graft fixation, and minimal immunosuppressive regime usage. The aim of this study is to explore the feasibility and efficacy of preparing porcine DMEK grafts by 2 techniques and investigate the graft ultrastructure. Methods Two mainstream techniques, mechanical stripping technique and liquid bubble technique, were modified to prepare the porcine DMEK grafts. In all, 40 corneas harvested from WZS‐pigs (aged 10‐12 months) were subjected to the techniques (20 corneas for each technique). The success rate, time consumption, and endothelial cell density (ECD) before and after preparation were recorded and compared between the 2 techniques. And the ultrastructure of the porcine DEMK graft was investigated by transmission electron microscope. In addition, 9 WZS‐pigs with different ages were sacrificed to explore the correlation between the thickness of Descemet’s membrane and porcine age. Results After modifying several technical details, the porcine DMEK grafts were successfully prepared by either mechanical stripping technique or liquid bubble technique, and the mark technique to distinguish the 2 sides of the graft was also explored. In all, 13 DMEK grafts (65%) were prepared successfully by the mechanical stripping technique, whereas 14 successful cases (70%) were prepared by the liquid bubble technique. The success rates between the 2 techniques showed no significant difference (P = .847). However, the mechanical stripping technique was significantly time‐consuming when compared with the liquid bubble technique (P &lt; .0001). The ECDs reduced significantly after preparation no matter what techniques were used (P &lt; .0001), but the ECD after the liquid bubble preparation was significantly higher than the ECD after mechanical stripping (P = .032). The ECD reduction positively correlated to the time consumption for both mechanical stripping technique (P = .0014, R2 = 0.621) and liquid bubble technique (P = .013, R2 = 0.412). The ultrastructure showed the graft was comprised of stromal residuals, non‐banded layer, and endothelial layer. Unlike human Descemet’s membrane (DM), anterior banded layer was not observed. The thickness of porcine DM increased with the age, and a significant positive correlation between them was found (P &lt; .0001, R2 = 0.949), and the predict equation was Y = 0.3764*X + 7.378 (Y indicates the thickness, whereas X indicates the age). Conclusions Porcine DMEK grafts could be prepared either by mechanical stripping technique or liquid bubble technique, and the liquid bubble technique seems superior over the mechanical stripping technique regarding time consumption and ECD preservation. Although there are several technical barriers to overcome, xeno‐DMEK might be a promising direction for future xeno‐corneal transplantation.</description><subject>Adult</subject><subject>Age</subject><subject>Aged</subject><subject>Animals</subject><subject>Cell density</subject><subject>Cornea</subject><subject>Corneal Diseases - immunology</subject><subject>Corneal Diseases - surgery</subject><subject>Corneal transplantation</subject><subject>Corneal Transplantation - methods</subject><subject>Descemet Membrane - immunology</subject><subject>Descemet’s membrane endothelial keratoplasty</subject><subject>Endothelial cells</subject><subject>ex‐vivo</subject><subject>Female</subject><subject>graft preparation</subject><subject>Graft rejection</subject><subject>Graft Survival - immunology</subject><subject>Humans</subject><subject>Immunosuppression</subject><subject>Investigations</subject><subject>liquid bubble technique</subject><subject>mechanical stripping technique</subject><subject>Middle Aged</subject><subject>porcine cornea</subject><subject>Preservation</subject><subject>Swine</subject><subject>Tissue and Organ Harvesting</subject><subject>Tissue Donors</subject><subject>Transplantation, Heterologous - methods</subject><subject>Transplants - immunology</subject><subject>Ultrastructure</subject><subject>Xenografts</subject><subject>xeno‐corneal transplantation</subject><issn>0908-665X</issn><issn>1399-3089</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kc1u1DAURi0EokNhwQsgS2zaRVo7jhOHXdUOP6LApkjdRXZyPeOS2KntTJtdH4Etz8Eb9UnwMIUFEndzpaujo_vpQ-glJUc0zfEt2COaF6R6hBaU1XXGiKgfowWpicjKkl_uoWchXBFCGBf8KdrL64qXBSUL9PNiDViDDEaZ3sQZS9th0Nq0sp2x03j0MEpv7AqPzrfGAj6D0MIA8f7uR8ADDMrLdAXbubiG3sgefwMvoxt7GZLw4OzT8uMhXnmpY8BqxvHG4Qjt2prrCcIbfGIx3N7ffd-YjcPGbiBEs5LROIu181hPcfKAU0aXoK3sOXqiZR_gxcPeR1_fLi9O32fnX959OD05z1rGWZUB7QqmqSCU5CUpuCiKlrUKmBSMl7wqlWIiTwehlapB5FXJOylZ2em66KBj--hg5x29274am8Gk6H2f8ropNDlhoiKcsiqhr_9Br9zkbfquySnNKWElZYk63FGtdyF40M3ozSD93FDSbJtsUsrmd5OJffVgnNQA3V_yT3UJON4BN6aH-f-m5nL5eaf8BVWBrVU</recordid><startdate>201809</startdate><enddate>201809</enddate><creator>Liu, Yang</creator><creator>Zhang, Jing</creator><creator>Zhang, Yingnan</creator><creator>Yin, Mingyang</creator><creator>Miao, Sen</creator><creator>Liang, Qingfeng</creator><creator>Pan, Zhiqiang</creator><general>Wiley Subscription Services, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7T5</scope><scope>H94</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-6854-0615</orcidid></search><sort><creationdate>201809</creationdate><title>The feasibility and efficacy of preparing porcine Descemet’s membrane endothelial keratoplasty (DMEK) grafts by two techniques: An ex‐vivo investigation for future xeno‐DMEK</title><author>Liu, Yang ; Zhang, Jing ; Zhang, Yingnan ; Yin, Mingyang ; Miao, Sen ; Liang, Qingfeng ; Pan, Zhiqiang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3537-e1d43f1801026045844c3cbe3a8356576bb382cbe8fbb9e82765daa36df94ded3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Adult</topic><topic>Age</topic><topic>Aged</topic><topic>Animals</topic><topic>Cell density</topic><topic>Cornea</topic><topic>Corneal Diseases - immunology</topic><topic>Corneal Diseases - surgery</topic><topic>Corneal transplantation</topic><topic>Corneal Transplantation - methods</topic><topic>Descemet Membrane - immunology</topic><topic>Descemet’s membrane endothelial keratoplasty</topic><topic>Endothelial cells</topic><topic>ex‐vivo</topic><topic>Female</topic><topic>graft preparation</topic><topic>Graft rejection</topic><topic>Graft Survival - immunology</topic><topic>Humans</topic><topic>Immunosuppression</topic><topic>Investigations</topic><topic>liquid bubble technique</topic><topic>mechanical stripping technique</topic><topic>Middle Aged</topic><topic>porcine cornea</topic><topic>Preservation</topic><topic>Swine</topic><topic>Tissue and Organ Harvesting</topic><topic>Tissue Donors</topic><topic>Transplantation, Heterologous - methods</topic><topic>Transplants - immunology</topic><topic>Ultrastructure</topic><topic>Xenografts</topic><topic>xeno‐corneal transplantation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liu, Yang</creatorcontrib><creatorcontrib>Zhang, Jing</creatorcontrib><creatorcontrib>Zhang, Yingnan</creatorcontrib><creatorcontrib>Yin, Mingyang</creatorcontrib><creatorcontrib>Miao, Sen</creatorcontrib><creatorcontrib>Liang, Qingfeng</creatorcontrib><creatorcontrib>Pan, Zhiqiang</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Xenotransplantation (Københaven)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liu, Yang</au><au>Zhang, Jing</au><au>Zhang, Yingnan</au><au>Yin, Mingyang</au><au>Miao, Sen</au><au>Liang, Qingfeng</au><au>Pan, Zhiqiang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The feasibility and efficacy of preparing porcine Descemet’s membrane endothelial keratoplasty (DMEK) grafts by two techniques: An ex‐vivo investigation for future xeno‐DMEK</atitle><jtitle>Xenotransplantation (Københaven)</jtitle><addtitle>Xenotransplantation</addtitle><date>2018-09</date><risdate>2018</risdate><volume>25</volume><issue>5</issue><spage>e12407</spage><epage>n/a</epage><pages>e12407-n/a</pages><issn>0908-665X</issn><eissn>1399-3089</eissn><abstract>Background Descemet’s membrane endothelial keratoplasty (DMEK) might be a promising technique for future xeno‐corneal transplantation due to its ultrathin graft, extremely low rejection occurrence, suture‐free graft fixation, and minimal immunosuppressive regime usage. The aim of this study is to explore the feasibility and efficacy of preparing porcine DMEK grafts by 2 techniques and investigate the graft ultrastructure. Methods Two mainstream techniques, mechanical stripping technique and liquid bubble technique, were modified to prepare the porcine DMEK grafts. In all, 40 corneas harvested from WZS‐pigs (aged 10‐12 months) were subjected to the techniques (20 corneas for each technique). The success rate, time consumption, and endothelial cell density (ECD) before and after preparation were recorded and compared between the 2 techniques. And the ultrastructure of the porcine DEMK graft was investigated by transmission electron microscope. In addition, 9 WZS‐pigs with different ages were sacrificed to explore the correlation between the thickness of Descemet’s membrane and porcine age. Results After modifying several technical details, the porcine DMEK grafts were successfully prepared by either mechanical stripping technique or liquid bubble technique, and the mark technique to distinguish the 2 sides of the graft was also explored. In all, 13 DMEK grafts (65%) were prepared successfully by the mechanical stripping technique, whereas 14 successful cases (70%) were prepared by the liquid bubble technique. The success rates between the 2 techniques showed no significant difference (P = .847). However, the mechanical stripping technique was significantly time‐consuming when compared with the liquid bubble technique (P &lt; .0001). The ECDs reduced significantly after preparation no matter what techniques were used (P &lt; .0001), but the ECD after the liquid bubble preparation was significantly higher than the ECD after mechanical stripping (P = .032). The ECD reduction positively correlated to the time consumption for both mechanical stripping technique (P = .0014, R2 = 0.621) and liquid bubble technique (P = .013, R2 = 0.412). The ultrastructure showed the graft was comprised of stromal residuals, non‐banded layer, and endothelial layer. Unlike human Descemet’s membrane (DM), anterior banded layer was not observed. The thickness of porcine DM increased with the age, and a significant positive correlation between them was found (P &lt; .0001, R2 = 0.949), and the predict equation was Y = 0.3764*X + 7.378 (Y indicates the thickness, whereas X indicates the age). Conclusions Porcine DMEK grafts could be prepared either by mechanical stripping technique or liquid bubble technique, and the liquid bubble technique seems superior over the mechanical stripping technique regarding time consumption and ECD preservation. Although there are several technical barriers to overcome, xeno‐DMEK might be a promising direction for future xeno‐corneal transplantation.</abstract><cop>Denmark</cop><pub>Wiley Subscription Services, Inc</pub><pmid>29756410</pmid><doi>10.1111/xen.12407</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0002-6854-0615</orcidid></addata></record>
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subjects Adult
Age
Aged
Animals
Cell density
Cornea
Corneal Diseases - immunology
Corneal Diseases - surgery
Corneal transplantation
Corneal Transplantation - methods
Descemet Membrane - immunology
Descemet’s membrane endothelial keratoplasty
Endothelial cells
ex‐vivo
Female
graft preparation
Graft rejection
Graft Survival - immunology
Humans
Immunosuppression
Investigations
liquid bubble technique
mechanical stripping technique
Middle Aged
porcine cornea
Preservation
Swine
Tissue and Organ Harvesting
Tissue Donors
Transplantation, Heterologous - methods
Transplants - immunology
Ultrastructure
Xenografts
xeno‐corneal transplantation
title The feasibility and efficacy of preparing porcine Descemet’s membrane endothelial keratoplasty (DMEK) grafts by two techniques: An ex‐vivo investigation for future xeno‐DMEK
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