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Research Background:The use of human biological valves referred to as allograft heart valves（AHV）is shown to have numerous improvements above conventional xenografts,significantly concerning hemodynamic reaction and durability.This allogeneic heart valves preservation necessitates meticulous attention to ensure that the cell integrity is preserved prior to clinical application.Recently,vitrification protocols were reported to preserve the matrix integrity of allograft valves and concomitantly result in low tissue immunogenicity.Trehalose has been shown to be involved in retaining cell viability after thawing in IFC protocols.Previous researches showed that nanoparticle encapsulated trehalose resulted in cryopreservation of mammalian cells with high viability and unblemished function.In practice,they are using a nanoparticle to encapsulate Trehalose and using it in the VS83 protocol results in minor ice crystallization.Hemocompatibility of cryopreserved heart valve has been a hurdle demanding the recipient to live on anticoagulants for their entire life.Calcification and inflammation significantly hinder the valve grafting due to hemocompatibility.Both these issues can be sorted out by tissue engineering which will take a long time to achieve it.Trehalose’s effect on hemocompatibility must be evaluated because it has been shown to be excellent at maintaining cell viability.Objective:This study aims to see whether Trehalose affects the allogeneic valves hemocompatibility in the VS83 procedure,precisely if the disaccharide will induce an immune reaction and decreases immune response after thawing.In addition,the research seeks to assess if nanoparticle-encapsulated Trehalose increases cell viability in IFC cardiovascular preservation techniques and maintains the extracellular matrix integrity.The primary theory is that Trehalose increases cell viability by vitrifying allogeneic heart valves and safeguards ECM structures.The second theory states that the absorption of Trehalose intracellularly results in no fundamental hematocompatibility involvement in allogeneic heart valves VS83 vitrification.Methods and Results:The preparation of hydroxyapatite nanoparticles was the initial step.The system of precipitation was acquired for apatite colloidal suspension.Animals were caught,and tissues were ethically isolated and grouped.Both samples were then assigned to 4 groups: Control group,CFC group,IFC group,and IFC+T group.Histological evaluation of the tissue was conducted,Extracellular matrix components were stained with modified Weigert staining,and reticular fibers were stained by Gomori ammonia method.To assess cell viability,the Alamar blue assay is currently used.Immunohistochemistry（IL-10 and TNFα）was conducted on the samples to determine their hemocompatibility.Neutrophil Elastase and Terminal Complement Complex have to be used as markers to determine hemocompatibility.H&E staining revealed that the IFC+T group formed fewer intracellular vacuoles and extracellular ice crystals in comparison to all other groups.The CFC group had higher cell viability than that of the IFC group,however the IFC+T group had the highest viability（70.96±2.53,P < 0.0001,n=6）.The levels of TNFα were lowest in both the IFC and IFC+T groups,and IL-10 expression was slightly lower in both the IFC and IFC+T groups,according to immunohistochemistry.Conclusion:Trehalose encapsulated by nanoparticles with VS83 formulation exhibited less ice formation.The morphology of tissues was not affected.Cell viability after thawing showed a noteworthy improvement in the preservation of tissues.This study additionally displays that integration of Trehalose did not elicit the creation of immunogenic and thrombogenic indicators,which indicates that there will be less hemocompatibility.