{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/81272"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/81272","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"Hypothermic Machine Perfusion of Composite Tissues","abstract":"Organ perfusion systems have successfully been applied to solid organ preservation and subsequent transplantation. However, their use in limb preservation for Vascularized Composite Tissue Allotransplantation (VCA) has yet to be thoroughly investigated. This thesis explores the potential for hypothermic machine perfusion in prolonging limb graft viability in a swine forelimb amputation model. The experiment was designed with the right and left forelimbs from the same pig randomly assigned to the treatment and control groups. Eighteen (18) limbs were procured from a local abattoir, vessels cannulated, and an initial flush of a modified phosphate buffered saline (PBS) solution was performed. Half of those limbs, assigned to the treatment group, were then preserved with continuous hypothermic machine perfusion for 12 hours. The perfusate was a PBS solution supplemented with 5% w/v dextrose. The remaining nine limbs, assigned to the control group, were placed into a plastic bag and kept at room temperature (ca. 20oC) for the entire duration of the experiment. Methylene blue was used to verify perfusion throughout limbs. Histopathological analysis revealed the presence of significantly greater deterioration of the perfused limbs compared to control. I concluded that PBS solution is not suitable for extended limb preservation. Inadequate perfusate volume and lack of solution replenishment resulted in metabolic waste build up, accelerating total organ damage. Continued research is needed in order to develop clinically relevant hypothermic machine perfusion devices capable of prolonged limb preservation.","abstract_html":"Organ perfusion systems have successfully been applied to solid organ preservation and subsequent transplantation. However, their use in limb preservation for Vascularized Composite Tissue Allotransplantation (VCA) has yet to be thoroughly investigated. This thesis explores the potential for hypothermic machine perfusion in prolonging limb graft viability in a swine forelimb amputation model. The experiment was designed with the right and left forelimbs from the same pig randomly assigned to the treatment and control groups. Eighteen (18) limbs were procured from a local abattoir, vessels cannulated, and an initial flush of a modified phosphate buffered saline (PBS) solution was performed. Half of those limbs, assigned to the treatment group, were then preserved with continuous hypothermic machine perfusion for 12 hours. The perfusate was a PBS solution supplemented with 5% w/v dextrose. The remaining nine limbs, assigned to the control group, were placed into a plastic bag and kept at room temperature (ca. 20oC) for the entire duration of the experiment. Methylene blue was used to verify perfusion throughout limbs. Histopathological analysis revealed the presence of significantly greater deterioration of the perfused limbs compared to control. I concluded that PBS solution is not suitable for extended limb preservation. Inadequate perfusate volume and lack of solution replenishment resulted in metabolic waste build up, accelerating total organ damage. Continued research is needed in order to develop clinically relevant hypothermic machine perfusion devices capable of prolonged limb preservation.","abstract_has_math":false,"creators":["Troutman Mr., Eric Christopher"],"institution":"Virginia Tech","degree_name":"Master of Science","degree_level":"masters","degree_discipline":"Mechanical Engineering","degree_department":"Mechanical Engineering","school":null,"contributors":[],"advisors":[],"committee_chairs":["Robertson, John L."],"committee_members":["Paul, Mark R.","Diller, Thomas E."],"year":2017,"date_issued":"2017-12-18","date_published":"2017-12-18","updated_at":"2026-07-22T22:19:48Z","subjects":["Engineering","Mechanical Engineering","Biomedical"],"languages":[],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["vt_gsexam:13693"],"render_values":[{"text":"vt_gsexam:13693","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/10919/81272","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.committeechair","label":"Committee Chair","values":["Robertson, John L."]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Paul, Mark R.","Diller, Thomas E."]},{"key":"dc:contributor.department","label":"Department","values":["Mechanical Engineering"]},{"key":"dc:creator","label":"Author","values":["Troutman Mr., Eric Christopher"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2017-12-19T09:00:18Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2017-12-19T09:00:18Z"]},{"key":"dc:date.issued","label":"Date","values":["2017-12-18"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Tech"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Mechanical Engineering"]},{"key":"thesis:degree_level","label":"Degree Level","values":["masters"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Virginia Polytechnic Institute and State University"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Engineering","Mechanical Engineering","Biomedical"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:rights","label":"Dc Rights","values":["In Copyright"]},{"key":"dc:rights.uri","label":"Rights URI","values":["http://rightsstatements.org/vocab/InC/1.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["vt_gsexam:13693"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10919/81272"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Organ perfusion systems have successfully been applied to solid organ preservation and subsequent transplantation. However, their use in limb preservation for Vascularized Composite Tissue Allotransplantation (VCA) has yet to be thoroughly investigated. This thesis explores the potential for hypothermic machine perfusion in prolonging limb graft viability in a swine forelimb amputation model. The experiment was designed with the right and left forelimbs from the same pig randomly assigned to the treatment and control groups. Eighteen (18) limbs were procured from a local abattoir, vessels cannulated, and an initial flush of a modified phosphate buffered saline (PBS) solution was performed. Half of those limbs, assigned to the treatment group, were then preserved with continuous hypothermic machine perfusion for 12 hours. The perfusate was a PBS solution supplemented with 5% w/v dextrose. The remaining nine limbs, assigned to the control group, were placed into a plastic bag and kept at room temperature (ca. 20oC) for the entire duration of the experiment. Methylene blue was used to verify perfusion throughout limbs. Histopathological analysis revealed the presence of significantly greater deterioration of the perfused limbs compared to control. I concluded that PBS solution is not suitable for extended limb preservation. Inadequate perfusate volume and lack of solution replenishment resulted in metabolic waste build up, accelerating total organ damage. Continued research is needed in order to develop clinically relevant hypothermic machine perfusion devices capable of prolonged limb preservation."]},{"key":"dc:description.abstractgeneral","label":"General Abstract","values":["Artificial circulatory systems, capable of maintaining organ viability outside the body, have successfully been used for kidney preservation prior to transplantation. However, this has not been thoroughly investigated for limb preservation. This thesis explores the potential for an artificial circulatory system to prolong the life of pig front limbs following amputation. Nine (9) pairs of limbs were used. From each pair, one limb was randomly assigned to the treatment group and the other to the control group. The limbs assigned to the treatment group were attached to a machine that continuously circulated a cold preservation solution for twelve (12) hours. A blue dye was used to verify that the solution was being circulated throughout the whole limb. The remaining nine (9) limbs were in the control group, placed in individual plastic bags and left at room temperature for the entire duration of the experiment. After the experiment, microscopic examination of the tissue revealed that there was greater deterioration of the treatment group limbs versus the control group. I concluded that the study’s preservation solution and its use, was not beneficial for limb preservation. Without a greater volume and replenishment of the solution, toxins were able to build up in the solution causing damage to the limbs. Continued research is needed to develop a circulatory system capable of extending the viability of a limb for transplantation."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Master of Science"]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["ETD"]},{"key":"dc:title","label":"Title","values":["Hypothermic Machine Perfusion of Composite Tissues"]}]}],"canonical_facts":{"dc:contributor.committeechair":["Robertson, John L."],"dc:contributor.committeemember":["Paul, Mark R.","Diller, Thomas E."],"dc:contributor.department":["Mechanical Engineering"],"dc:creator":["Troutman Mr., Eric Christopher"],"dc:date.accessioned":["2017-12-19T09:00:18Z"],"dc:date.available":["2017-12-19T09:00:18Z"],"dc:date.issued":["2017-12-18"],"dc:description.abstract":["Organ perfusion systems have successfully been applied to solid organ preservation and subsequent transplantation. However, their use in limb preservation for Vascularized Composite Tissue Allotransplantation (VCA) has yet to be thoroughly investigated. This thesis explores the potential for hypothermic machine perfusion in prolonging limb graft viability in a swine forelimb amputation model. The experiment was designed with the right and left forelimbs from the same pig randomly assigned to the treatment and control groups. Eighteen (18) limbs were procured from a local abattoir, vessels cannulated, and an initial flush of a modified phosphate buffered saline (PBS) solution was performed. Half of those limbs, assigned to the treatment group, were then preserved with continuous hypothermic machine perfusion for 12 hours. The perfusate was a PBS solution supplemented with 5% w/v dextrose. The remaining nine limbs, assigned to the control group, were placed into a plastic bag and kept at room temperature (ca. 20oC) for the entire duration of the experiment. Methylene blue was used to verify perfusion throughout limbs. Histopathological analysis revealed the presence of significantly greater deterioration of the perfused limbs compared to control. I concluded that PBS solution is not suitable for extended limb preservation. Inadequate perfusate volume and lack of solution replenishment resulted in metabolic waste build up, accelerating total organ damage. Continued research is needed in order to develop clinically relevant hypothermic machine perfusion devices capable of prolonged limb preservation."],"dc:description.abstractgeneral":["Artificial circulatory systems, capable of maintaining organ viability outside the body, have successfully been used for kidney preservation prior to transplantation. However, this has not been thoroughly investigated for limb preservation. This thesis explores the potential for an artificial circulatory system to prolong the life of pig front limbs following amputation. Nine (9) pairs of limbs were used. From each pair, one limb was randomly assigned to the treatment group and the other to the control group. The limbs assigned to the treatment group were attached to a machine that continuously circulated a cold preservation solution for twelve (12) hours. A blue dye was used to verify that the solution was being circulated throughout the whole limb. The remaining nine (9) limbs were in the control group, placed in individual plastic bags and left at room temperature for the entire duration of the experiment. After the experiment, microscopic examination of the tissue revealed that there was greater deterioration of the treatment group limbs versus the control group. I concluded that the study’s preservation solution and its use, was not beneficial for limb preservation. Without a greater volume and replenishment of the solution, toxins were able to build up in the solution causing damage to the limbs. Continued research is needed to develop a circulatory system capable of extending the viability of a limb for transplantation."],"dc:description.degree":["Master of Science"],"dc:format.medium":["ETD"],"dc:identifier.other":["vt_gsexam:13693"],"dc:identifier.uri":["http://hdl.handle.net/10919/81272"],"dc:publisher":["Virginia Tech"],"dc:rights":["In Copyright"],"dc:rights.uri":["http://rightsstatements.org/vocab/InC/1.0/"],"dc:subject":["Engineering","Mechanical Engineering","Biomedical"],"dc:title":["Hypothermic Machine Perfusion of Composite Tissues"],"dc:type":["Thesis"],"thesis:degree_discipline":["Mechanical Engineering"],"thesis:degree_level":["masters"],"thesis:degree_name":["Master of Science"],"thesis:institution_name":["Virginia Polytechnic Institute and State University"]},"updated_at":"2026-07-22T22:19:48Z"}