Search Results (186)

Background/Objectives: A novel treatment of absolute uterine factor infertility is uterus transplantation. In preparation for human surgery, autotransplantation was performed in a sheep model to assess ischemia-reperfusion injury of the uterine wall. Methods: Seven multiparous ewes underwent live-donor uterus autotransplantation; in six, the procedure was completed successfully. Tissue blocks of complete uterine wall, endometrium, and myometrium were obtained at four predefined time points: native (baseline), after 1 h of cold ischemia, after 30 min of warm ischemia, and after 30 min of reperfusion. Samples were analyzed by differential scanning calorimetry and routine hematoxylin–eosin histology. Results: Histology demonstrated preserved epithelial, glandular, and stromal structures, with only minimal, reversible changes that increased with the ischemic duration. Differential scanning calorimetry confirmed alterations in thermal stability: in the uterine wall and myometrium, the calorimetric enthalpy decreased from baseline (3.40 ± 0.53 J/g) to reperfusion (2.62 ± 0.22 J/g), indicating structural loosening; in contrast, the endometrium calorimetric enthalpy slightly increased, suggesting greater flexibility and less susceptibility to ischemia-reperfusion injury. Conclusions: In this preliminary study, differential scanning calorimetry proved to be an effective and sensitive method for detecting early structural alterations in the uterine wall that could negatively impact post-transplant function. Cold and warm ischemia did not cause irreversible damage within a two-hour time frame, supporting the feasibility of short-term preservation in uterus transplantation. The myometrium demonstrated more significant vulnerability than the endometrium, which highlights the necessity of protective strategies to preserve smooth muscle integrity during transplantation. Full article

Machine perfusion technologies have redefined the landscape of organ preservation by enabling not just static cold storage, but graft optimization and assessment with the opportunity for additional therapeutic interventions. Preservation solutions, traditionally developed for static cold storage, are now being adapted for use in dynamic perfusion platforms. The optimal composition for machine perfusion remains unclear as we shift to creating biologically intelligent platforms tailored to mitigate ischemia–reperfusion injury. This review presents a mechanistic framework for understanding organ preservation through the lens of shared vulnerabilities, particularly: mitochondrial dysfunction, endothelial barrier breakdown, and the activation of inflammatory cascades. We discuss the evolution of classical preservation solutions, the rationale for redox-targeted and endothelial-stabilizing additives, and the promise of modular approaches adaptable to both static cold storage and machine perfusion. By integrating recent preclinical insights, systems biology, and emerging clinical trials, we outline the path toward unified, precision-preservation strategies capable of expanding the donor pool and improving transplant outcomes. Full article

Background/Objectives: The shortage of liver grafts remains a major challenge in transplantation. Full-left-full-right (FLFR) split liver transplantation (SLT) expands the donor pool by providing two grafts for small adult recipients. However, prolonged cold ischemia time (CIT) and ischemia-reperfusion injury (IRI) limit its success. Methods: We report a case of FLFR SLT utilizing ex situ dual hypothermic oxygenated machine perfusion (DHOPE) to mitigate IRI and enhance graft viability. A brain-dead donor’s liver was split under continuous DHOPE, followed by simultaneous transplantation into two adult recipients. Results: Both recipients exhibited stable graft function at one-year follow-up. DHOPE effectively reduced CIT and optimized postoperative recovery, with no major complications beyond Clavien–Dindo Grade IIIb. Conclusions: This is the first reported FLFR SLT using ex situ DHOPE for two adult recipients, demonstrating its feasibility in reducing CIT and improving outcomes. Machine perfusion may become a standard in FLFR SLT. Full article

Background/Objectives: APOL1 renal-risk variants (RRVs) are of increasing relevance to kidney disease and transplant outcomes. It is currently understood that the presence of RRVs in donors negatively impacts kidney allograft survival in an autosomal recessive pattern of inheritance. Less well known is the interplay between ischemia and alternative allograft preservation methods, such as normothermic machine perfusion (NMP), on APOL1 gene expression. To investigate this, we examined the effects of APOL1 RRVs on APOL1 gene expression in ischemic donor kidneys and compared the differences in cytokine and APOL1 expression patterns between the alternative preservation methods, static cold storage (CS) and NMP. Methods: Non-utilized deceased donor kidney pairs from donors of African ancestry were procured from Mid-America Transplant after being deemed unsuitable for kidney transplant. Samples were collected from each donor kidney pair and DNA was extracted for APOL1 genotyping. APOL1 RRVs G1 (rs73885319) (rs60910145) and G2 (rs71785313) were identified by Sanger sequencing. From each pair, one kidney underwent 6 h NMP (n = 3) and the contralateral kidney 6 h of CS (n = 3) following the initial CS. Renal perfusion and biochemical, and histologic parameters were recorded. NMP was directly compared with CS using paired donor kidneys using NMP with allogeneic red blood cells, followed by assessment of perfusion, biochemical, and histologic parameters, in addition to gene expression. Results: Donor genotyping identified kidney pairs as heterozygous for the G1 RRV (G1/G0), homozygous for the G0 allele (G0/G0), and homozygous for the G2 RRV (G2/G2), respectively. All kidneys were successfully reperfused, with mRNA transcript levels of APOL1-related genes subsequently measured. Significant differences in APOL1 gene expression were observed among all three groups of kidneys. In paired kidneys from baseline to hour 6 of NMP, mRNA expression varied significantly between G1/G0 and G2/G2 homozygous pairs (p = 0.002) as well as between the G0/G0 and G2/G2 pairs (p = 0.002). APOL1 expression shifted by a significantly higher-fold change of 2.4 under NMP conditions in the G2/G2 genotype (p < 0.001). The inflammatory cytokine marker IFN-γ was also significantly upregulated in the G2/G2 genotype kidney, in both CS and NMP groups (p = 0.001). Other related genes such as KIM-1 were upregulated by a change of 3.9-fold in the NMP group for the G2/G2 kidney. Conclusion: Donor kidney pairs with the high-risk APOL1 genotype, especially G2/G2, show increased APOL1 expression and inflammation, particularly under NMP conditions. NMP enables detection of genotype-specific molecular changes in an ischemic reperfusion injury model, supporting its potential to improve donor kidney assessment before transplantation. Full article

Liver transplantation is commonly used for end-stage liver disease, but the demand for organs exceeds the supply, leading to the use of expanded criteria donors (ECDs). Organs from ECDs, especially from donors after circulatory death (DCD), encounter challenges like increased ischemia damage. Biomarkers, especially oxidative stress markers, may provide valuable insights for understanding and monitoring post-transplant events. Here, we highlight the unique value of organ preservation solution (OPS) as a non-invasive and early source of redox biomarkers, directly reflecting graft status during critical cold storage. This study investigated oxidative stress in 74 donated livers using OPS samples collected after cold storage, and also liver biopsies obtained before and after storage. We measured lipid peroxidation, protein carbonylation, DNA oxidation, and total antioxidant capacity from OPS, and performed gene expression analysis of liver biopsies. Oxidative stress markers differed based on donation type, with higher lipid peroxidation in DCD samples compared with donation after brain death (18.51 ± 2.77 vs. 11.03 ± 1.31 nmoles malondialdehyde (MDA)/mg protein; p = 0.049). Likewise, oxidative damage markers were associated with clinical outcomes: lipid peroxidation was increased in patients who developed biliary complications (21.86 ± 5.91 vs. 11.97 ± 1.12 nmol MDA/mg protein; p = 0.05), and protein carbonylation was elevated in those experiencing acute rejection (199.6 ± 22.02 vs. 141.6 ± 15.94 nmol carbonyl/mg protein; p = 0.005). Moreover, higher protein carbonylation levels showed a trend toward reduced survival (p = 0.091). Transcriptomic analysis revealed overexpression of genes associated with reactive oxygen species production in DCD livers. A predictive model for acute rejection integrating OPS biomarkers with clinical variables achieved 83% accuracy. Hence, this study underscores the importance of assessing oxidative stress status in preservation fluid as a biomarker for evaluating liver transplant outcomes and highlights the need for validation in larger, independent cohorts. Full article

Background and Objectives: Kidney transplantation remains the gold-standard treatment for end-stage renal disease (ESRD). For deceased donor transplantation, optimal allograft preservation represents a critical determinant of success. While static cold storage (SCS) has been the historical standard, hypothermic machine perfusion (HMP) has emerged in recent decades as a technologically advanced alternative. However, comparative data from smaller-volume centers utilizing exclusively donation after brain death (DBD) donors remain scarce. Materials and Methods: This retrospective single-center study included 94 patients who received kidney transplants from deceased DBD donors between January 2018 and December 2024. We employed a paired kidney study design where one kidney from each donor was preserved using HMP (LifePort, set at 30/20 mm Hg pressure), while the contralateral kidney was stored in SCS. Parameters compared were creatinine concentration in recipient serum after transplantation, DGF, acute rejection and hospital stay. Results: The HMP group had a significantly longer cold ischemia time (CIT) (18.09 ± 5.91 h, range: 6.5–34.0 h) compared to the SCS group (12.36 ± 5.18 h, range: 4.0–23.0 h; p < 0.005). The DGF rate was significantly lower in the HMP group (4.3%) than the SCS group (25.5%) (p = 0.004). HMP was also associated with a shorter mean hospitalization (11.81 vs. 15.66 days, p = 0.008) and superior early graft function, particularly in kidneys with CIT ≥ 18 h, which showed significantly lower serum creatinine at day 14 (124.48 vs. 164.89 µmol/L, p = 0.036). Conclusions: HMP usage in kidney transplantation decreased the possibility for DGF in DBD donors and shortened the post-op hospitalization time. It is a feasible method for kidney storage before transplantation even in a case of prolonged CIT. Full article

Background and Objectives: The impact of sarcopenia and myosteatosis on renal transplantation (RT) outcomes has yet to be explained, certainly due to differences in assessment methods. The role of visceral adiposity is also not clearly defined. This retrospective study aimed to evaluate pretransplant body composition—including sarcopenia, myosteatosis, and visceral adiposity ratio (VSR)—using computed tomography (CT) and analyze their relationship with short- and long-term graft outcomes. Materials and Methods: A total of 94 patients who underwent RT between 2019 and 2023 and had pretransplant non-contrast abdominal CT scans were included. Skeletal muscle area (SMA) was assessed at the L3 vertebral level, including multiple muscle groups. Sarcopenia was defined by a low skeletal muscle index (SMI), while myosteatosis was defined by high intramuscular adipose tissue content (IMAC). Visceral adiposity was evaluated by the visceral-to-subcutaneous adipose tissue ratio (VSR). These parameters were compared with post-transplant outcomes. Results: The mean age was 42.69 ± 12.47 years, with 54.3% male patients. High IMAC was significantly associated with early graft failure (p = 0.026), delayed graft function (p = 0.005), death-censored graft failure (p = 0.036), and overall graft failure (p = 0.047). One-year mortality was also higher in the high IMAC group (14.8% vs. 0.0%, p = 0.012). SMI and VSR were not significantly associated with outcomes. Myosteatosis emerged as a significant risk factor in univariate analysis but was not independently predictive in multivariate analysis. Among the established risk factors identified in the study, recipient age was found to be a significant predictor for overall graft failure, donation type (cadaveric vs. living) for death-censored graft failure, and cold ischemia time for delayed graft function (OR: 1.068, 95% CI: 1.001–1.141, p = 0.049; OR: 147.7, 95% CI: 2.1—10,427.0, p = 0.021; OR: 1.003, 95% CI: 1.001–1.006, p = 0.023). Conclusions: Myosteatosis correlates with worse graft outcomes and higher mortality, but its independent prognostic value requires further investigation. Full article

Background: Static cold storage is the standard method of kidney preservation following donation after circulatory death (DCD). A previous study on rodent models demonstrated the efficacy of storing DCD kidneys at 22 °C in an extracellular-type solution (ETK). We evaluated the efficacy of storing warm-ischemic kidneys at 22 °C in MHC-inbred miniature swine. Methods: After 2 h warm ischemia, the kidneys were preserved in ETK for one hour at either 4 °C or 22 °C and then subjected to ex vivo normothermic machine perfusion (NMP) for 2 h (n = 3 in each group). The same warm-ischemic kidneys, preserved in ETK (n = 3 in each group) or intracellular-type solution (UW; n = 2 in each group) at either 4 °C or 22 °C, were transplanted into MHC-matched recipients. Results: Compared with kidneys preserved at 4 °C, those preserved at 22 °C showed significantly better physiological and metabolic indices during ex vivo NMP. Furthermore, renal function was significantly higher in transplanted kidneys, and graft biopsies on postoperative day 4 showed more localized necrosis in the renal tubules when kidneys were stored at 22 °C. In contrast, recipient animals with kidneys stored in UW solution did not survive for more than 7 days. Conclusions: Two-hour warm-ischemic kidneys from miniature swine showed improved preservation at 22 °C than at 4 °C when an extracellular-type solution was used. Full article

Background: The perfusion of viscera, kidney, and spinal cord represents one of the main concerns during open repair (OR) of Thoraco-Abdominal Aortic Aneurisms (TAAAs). Passive shunting (PS) has been historically used for intraoperative distal aortic perfusion but has been progressively replaced almost entirely by partial left-sided heart or total cardiopulmonary bypass with extra-corporeal circulation (ECC). Despite several advantages of these methods, PS still has potential in mitigating some drawbacks of long extracorporeal circuits connected with centrifugal or roller pumps, such as the need for cardiac and great vessels cannulation, priming and large intravascular fluid volume shifts, high heparin dose, immunosuppressive effects, and systemic inflammatory response syndrome. Methods: This study prospectively analyzed data of a cohort of patients who underwent TAAA OR using a PS in a single institution. Outcomes of interest were mortality, rate of mesenteric, renal and spinal cord ischemia, cardiac complications, and intraoperative hemodynamic stability achieved in this setting. Our institutional bundle and a comprehensive literature review about the different configurations and applicability of PS for TAAA OR is also reported. The search was performed based on three databases (PubMed, EMBASE, and Cochrane Library) by two independent reviewers (LS and AA) from inception to 31 December 2023, and the reported clinical results (visceral, renal, and spinal cord complications and mortality) using PS during TAAAs OR were analyzed. Results: Between March 2021 and December 2023, 51 TAAA repairs were performed and eleven patients (n = 8, 73% male; mean age 67 years, range 63–79) were operated using a PS for a total of one (9%) type I, one (9%) type II, two (18%) type III, five (45%) type IV, and two (18%) type V TAAA. In our early experience, PS was indicated for limited staff resources during the COVID-19 pandemic to treat five non-deferable cases. The sixth and seventh patients were selected for PS as they already had a functioning axillo-bifemoral bypass that was used for this purpose. For the most recent cases, PS was chosen as the primary perfusion method according to a score based on clinical and anatomical factors with ECC as a bailout strategy. Selective renal perfusion with cold (4 °C) Custodiol solution was the method of choice for renal protection in all cases while antegrade perfusion of the coeliac trunk and the superior mesenteric artery was assured by PS through a loop graft (8–10mm) proximally anastomosed to the axillary artery (10 patients, 90.9%) or the descending thoracic aorta (one patient, 9%) and distally anastomosed to the infrarenal aorta (3), common iliac (3), or femoral vessels (5). In-hospital mortality was 9% as one patient died on the 10th postoperative day from mesenteric ischemia following hemodynamic instability; permanent spinal cord ischemia rate was 0% and the rate of AKI stage 3 was 9% (one patient). Bailout shifting to ECC was never required. No cardiac complications, nor a significant increase in serum CK-MB were reported in any patient. No prolonged severe intraoperative hypotension episodes (Mean Arterial Pressure < 50 mmHg) were assessed using the Software Acumen Analytics (Edwards LifeSciences, Irvine CA, USA). No peri-operative coagulopathy nor major bleeding was reported. Conclusions: Our experience showed satisfactory outcomes with the use of PS in specifically selected cases. Current data indicate that PS may represent an alternative to ECC techniques during TAAAs OR in high volume centers where assisted extracorporeal circulation could eventually be applied as a bailout strategy. However, due to the small sample size of this and previously published series, more data are needed to clearly define the potential role of such approach during TAAA OR. Full article

The precipitation of cryoglobulins, serum immunoglobulins, below 37 °C defines the clinical cryoglobulinemic syndrome, a systemic vasculitis usually characterized by purpura, weakness, and arthralgia. In most cases, this condition is associated with chronic infection by the hepatitis C virus (HCV) and can evolve into B-cell dysregulation and malignancies. The current literature on non-HCV-associated cryoglobulinemia is very limited, and little is known about the immunological and serological profile of affected patients. The cryoglobulinemic syndrome not associated with HCV infection is often found concomitantly with other infections, autoimmune diseases, and B-cell lymphoproliferative disorders. The cryoprecipitation of fibrinogen has been described as a rare disorder, perhaps underestimated and not fully understood, causing thrombotic occlusion and ischemia in different rheumatic disorders. Cold temperature plays a pathogenetic role in autoimmune hemolytic anemias, in which the presence of cold agglutinins produced by B cells at the lymphoplasmacytic cell stage may promote agglutination of red blood cells in the coldest parts of the circulation, even at mild room temperatures, undergoing hemolysis. Laboratory methods for the detection and quantification of cryoproteins are downright critical, and their concurrent detection is pivotal for the diagnosis. In this review, we summarize the clinical involvement of cryoglobulins, cryofibrinogen, and cold agglutinins in non-HCV autoimmune diseases, underlining the crucial steps of the most employed analytic methods. Full article

Introduction: Liver transplantation remains the definitive treatment for end-stage liver disease but faces critical challenges including organ shortages and preservation difficulties, particularly with extended criteria donor (ECD) grafts. Hypothermic machine perfusion (HMP) represents a promising alternative to traditional static cold storage (SCS). Methods: This retrospective study analyzed outcomes from 62 liver transplant recipients between 2016 and 2025, comparing 8 grafts preserved by HMP using the Liver Assist^® system and 54 grafts preserved by SCS. Parameters assessed included postoperative complications, hemodynamic stability, ischemia times, and survival outcomes. Results: HMP significantly reduced surgical (0% vs. 75.9%, p = 0.01) and biliary complications (0% vs. 34.4%, p = 0.004), improved hemodynamic stability post-reperfusion (∆MAP%: 1 vs. 21, p = 0.006), and achieved superior one-year survival rates (100% vs. 84.4%). Despite longer ischemia periods, grafts treated with HMP exhibited fewer adverse effects from ischemia-reperfusion injury. Discussion: These findings highlight the substantial benefits of HMP, particularly in improving graft quality from marginal donors and reducing postoperative morbidity. Further adoption of this technology could significantly impact liver transplantation outcomes by expanding the viable donor pool. Conclusions: The study underscores the effectiveness of hypothermic machine perfusion (HMP) as a superior preservation method compared to traditional static cold storage (SCS), HMP appears to be associated with improved short-term outcomes in liver transplantation. By substantially reducing postoperative complications and enhancing graft viability, HMP emerges as a pivotal strategy for maximizing the use of marginal donor organs. Further research and broader clinical implementation are recommended to validate these promising results and to fully harness the potential of HMP in liver transplantation. Full article

Organ shortage is a major challenge in transplantation, prompting the use of extended criteria donor grafts. These require improved preservation techniques and reliable methods to assess graft function. This study aimed to evaluate changes in the kidney metabolome following three preservation methods: normothermic ex vivo kidney perfusion (NEVKP), hypothermic machine perfusion (HMP) and static cold storage (SCS) in porcine autotransplant models. A chemical biopsy allowed minimally invasive sampling of metabolites, which were analyzed using liquid chromatography coupled with high-resolution mass spectrometry. The results highlighted metabolites affected by ischemia and oxidative stress in donor kidneys, as well as changes specific to each preservation method. Differences were observed immediately after transplantation and reperfusion and several days post-surgery. NEVKP was associated with the activation of physiological anti-oxidative and anti-inflammatory mechanisms, suggesting potential protective effects. However, some metabolites had dual roles, which may influence future graft treatment designs. HMP and SCS, while reducing energy demand in cells, also limit physiological repair mechanisms. These findings provide a basis for improving graft assessment and organ preservation, with chemical biopsy serving as both a tool for discovery and a potential diagnostic method for monitoring graft quality. Full article

Background: Hypothermia has been previously reported to ameliorate acute renal injury induced by ischemia–reperfusion injury (IRI). However, its protective effects against subsequent renal fibrosis remain unclear. Objectives: The aim of this study was to determine whether hypothermia provides protection against renal ischemia–reperfusion injury (IRI), and to elucidate the molecular mechanisms involved. Methods: We used a model of renal fibrosis after ischemia–reperfusion injury in mice. C57BL/6 mice were divided into the following groups: control mice and ischemia–reperfusion injury mice (at 37 °C and at 32 °C). Their kidneys were harvested on day 1, day 3, and day 7 after IRI. The molecular mechanisms were evaluated. Results: The blood urea nitrogen (BUN) levels, serum creatinine (s-Cr) levels, and the histologic renal injury scores were significantly lower in the 32 °C IRI group than in the 37 °C ischemia–reperfusion injury group. In the hypothermic IR group, TGF-β and α-SMA were significantly decreased, while the PGC-1α level was significantly increased. Cold preparation increased the PGC-1α levels in HK2 cells. In TGF-β-treated HK2 cells, cold preparation decreased α-SMA and collagen IV levels. In addition, siPGC-1α in HK2 cells increased α-SMA and collagen IV, despite cold preparation. Conclusions: Hypothermia attenuates renal function deterioration and renal fibrosis in renal IRI mice kidneys. PGC-1α may play a role in hypothermic protection in renal fibrosis after IRI. Full article

Background: Acute rejection (AR) in kidney transplant (KT) recipients remains a significant challenge for short- and long-term graft survival even in the most recent years characterized by extended criteria donors and older and more comorbid recipients. Methods: We analyzed risk factors and outcomes of AR in 339 KT recipients treated at St. Orsola-Malpighi Hospital, Bologna (Italy), between 1 January 2019 and 31 December 2021. Demographic, immunological, and transplant data (type, cold ischemia time, complications) were recorded with a follow-up period of up to 24 months. Key outcomes included estimated glomerular filtration rate (eGFR), 24 h proteinuria, delayed graft function (DGF), biopsy-proven AR, and graft loss. Results: During the first year after transplant, 57 AR episodes occurred: 19 antibody-mediated rejections (AMR), 18 borderline T cell-mediated rejections (TCMR), 18 TCMR, 2 mixed AMR/TCMR, and 11 graft losses. AR was linked to older donor age (59.9 ± 12.8 vs. 55.5 ± 15.1, p = 0.040), longer cold ischemia time (690 vs. 570 min, p = 0.044), higher DGF rates (61.40% vs. 39.57%, p = 0.002), and lower eGFR (39 vs. 52 mL/min, p = 0.003). AR was consistently prevalent in patients who underwent an AB0-incompatible (AB0-i) transplant (8.8% vs. 2.5%, p = 0.020). HLA matching was strongly associated with a reduced risk of AMR (HLA-DR: OR 0.35, HLA-A: OR 0.33, HLA-C: OR 0.35), while DGF was linked to a higher risk (OR 4.04). TCMR risk was associated with donor age (OR 1.05). The development of post-transplant donor-specific antibodies (DSAs) at 24 months showed no significant association with AR (AMR: p = 0.769; TCMR: p = 0.938). The decline in eGFR over time (24 months) did not differ between patients with and without AR (difference, −0.69 mL/min/year; Standard Error, 0.92; p = 0.452). Similarly, 24 h proteinuria change over time did not differ between patients with and without AR (difference, −0.12 g/24 h; Standard Error, 0.28; p = 0.657). Conclusions: Understanding the risk factors of AR is crucial to identifying KTs at more risk of rejection and to guiding targeted therapeutic decisions. In the most recent era of extended criteria donors and more vulnerable recipients, early diagnosis and prompt and tailored treatment of AR play a critical role in stabilizing renal function over time. Full article

Tissue substitution and graft transplantation are currently the best treatment options for patients suffering from severe heart diseases. However, the limited availability of donors and the restricted durability of tissues applied in cardiovascular treatments result in a constraint on applicability and a suboptimal therapeutic approach that is still not fully resolved. There are multiple ways to preserve heart tissue grafts, and the choice of method is solely dependent upon the nature and complexity of the tissue and the length of storage. The conventional cold storage method provides the base to nearly all of the preservation protocols for short- and long-term storage. Short-term storage methods frequently rely on designing preserving solutions to protect the graft against warm and cold ischemia at the temperature above freezing point. As ice-nucleation is the major notorious phenomenon during graft preservation, the modern era of research is focusing on developing ice-free preservation techniques, termed vitrification. However, despite the promising outcomes of vitrification, there are several recognized hurdles required to be overcome to build a biobank of heart grafts for an extended period of time. Besides tissue deterioration due to extreme cold temperature, there is another extreme phenomenon of tissue rejection mainly caused by the presence of cellular antigens. The modern approach of decellularization has the potential to minimize the chances of tissue rejection by removing the cells and providing a structural support and sustained biochemical signal via keeping the extracellular matrix of the graft intact. In conclusion, both nano-warming and decellularization are the leading approaches that have great potential to store the graft tissue in its optimal form via keeping its viability safe for a longer time and extending its applicability. This review article outlines a variety of approaches for the preservation and bioengineering of tissue to fulfill the need for the availability of on-shelf long-lasting grafts both in clinical and laboratory setups. Full article