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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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 |
doi_str_mv | 10.1111/xen.12407 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2038705137</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2038705137</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3537-e1d43f1801026045844c3cbe3a8356576bb382cbe8fbb9e82765daa36df94ded3</originalsourceid><addsrcrecordid>eNp1kc1u1DAURi0EokNhwQsgS2zaRVo7jhOHXdUOP6LApkjdRXZyPeOS2KntTJtdH4Etz8Eb9UnwMIUFEndzpaujo_vpQ-glJUc0zfEt2COaF6R6hBaU1XXGiKgfowWpicjKkl_uoWchXBFCGBf8KdrL64qXBSUL9PNiDViDDEaZ3sQZS9th0Nq0sp2x03j0MEpv7AqPzrfGAj6D0MIA8f7uR8ADDMrLdAXbubiG3sgefwMvoxt7GZLw4OzT8uMhXnmpY8BqxvHG4Qjt2prrCcIbfGIx3N7ffd-YjcPGbiBEs5LROIu181hPcfKAU0aXoK3sOXqiZR_gxcPeR1_fLi9O32fnX959OD05z1rGWZUB7QqmqSCU5CUpuCiKlrUKmBSMl7wqlWIiTwehlapB5FXJOylZ2em66KBj--hg5x29274am8Gk6H2f8ropNDlhoiKcsiqhr_9Br9zkbfquySnNKWElZYk63FGtdyF40M3ozSD93FDSbJtsUsrmd5OJffVgnNQA3V_yT3UJON4BN6aH-f-m5nL5eaf8BVWBrVU</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2112103613</pqid></control><display><type>article</type><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><source>MEDLINE</source><source>Wiley Online Library Journals Frontfile Complete</source><creator>Liu, Yang ; Zhang, Jing ; Zhang, Yingnan ; Yin, Mingyang ; Miao, Sen ; Liang, Qingfeng ; Pan, Zhiqiang</creator><creatorcontrib>Liu, Yang ; Zhang, Jing ; Zhang, Yingnan ; Yin, Mingyang ; Miao, Sen ; Liang, Qingfeng ; Pan, Zhiqiang</creatorcontrib><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 < .0001). The ECDs reduced significantly after preparation no matter what techniques were used (P < .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 < .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 & Sons A/S. Published by John Wiley & Sons Ltd.</rights><rights>2018 John Wiley & 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 < .0001). The ECDs reduced significantly after preparation no matter what techniques were used (P < .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 < .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 < .0001). The ECDs reduced significantly after preparation no matter what techniques were used (P < .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 < .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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