P20-A129†An innovative wound healing method reveals a donor and post-mortem time-dependent regeneration of corneal epithelium.

PURPOSE: The aim of this study was to establish and optimize a new and reproducible epithelial wound healing model on human corneas. This assay was used to study the kinetics of epithelial regeneration following a chemical injury. METHODS: Thirty (n=30) human corneas unsuitable for transplant were used for the experiments. Corneas were cultured in Storagix medium (FBOV) at 31°C. Epithelial integrity before the beginning of the experiments (pre-wound) was assessed using the vital dyes trypan blue (TB, TB-S 0.25%, AL.CHI.MI.A. srl) and sodium fluorescein (Fluo). 1-heptanol soaked paper disks (6 mm) were applied in the centre of the corneas for 1' to trigger a chemical damage at the epithelial layer. Afterwards, sodium fluorescein and TB stainings were repeated to quantify the damaged area and to monitor healing progression. The damaged area (mm2) was calculated for each time point with Fiji software. Wound healing rate (HR, mm2/die) was calculated for both Fluo (HRF) and TB (HRTB) measurements using the previously described formula:Arithmetical averages (HRFAVG and HRTBAVG) of HRs were calculated and correlated by Pearson correlation coefficient with the following donor's parameters: age, sex, post-mortem time (PMT, time between death and tissue procurement), stromal defects, septicaemia, body temperature, diabetes. RESULTS: The execution of the heptanol wounding is highly reproducible, as highlighted by Fluo and TB staining. The average time for full recovery from wounding was 3,8 ± 0,41 days for Fluo and 3,5 ± 0,63 days for TB. Fluo and TB stainings are interchangeable as they significantly correlate (Pearson correlation coefficient = 0.630; p>0.05). A negative linear correlation was observed between HR and PMT (HRFAVG: corrected R2: 0.243, p = 0.003; HRTBAVG: corrected R2: 0,132, p = 0.028), but not with the other donors' parameters. CONCLUSION: Our wound/healing model might be of great interest for studies of epithelial regeneration kinetics and validation of drugs for the treatment of ocular defects. The inverse correlation between PMT and HR provides valuable insights for scientists investigating the regenerative properties of the corneal epithelium, as well as for eye bank personnel aiming to preserve the regenerative potential of corneal epithelium.

Presence of SARS-CoV-2 RNA in human corneal tissues donated in Italy during the COVID-19 pandemic.

OBJECTIVE: To analyse corneal tissues from asymptomatic donors with a postmortem nasopharyngeal swab tested positive for the presence of SARS-CoV-2 RNA, and therefore, understand the role that corneal transplantation may have in viral transmission. METHODS AND ANALYSIS: Between March 2020 and October 2021, 101 corneas (out of 8154 collected in Italy) from 51 donors (out of a total of 4155 Italian donors) positive for SARS-CoV-2 after postmortem nasopharyngeal swab tests were analysed for the presence of SARS-CoV-2 RNA through real-time RT-PCR. When available, the corneal tissue storage media were also assessed. Corneas and/or storage media with confirmed presence of SARS-CoV-2 RNA were further investigated by isolating SARS-CoV-2 virions, which were used to infect VeroE6 target cells. RESULTS: Only N=4 corneas and/or storage media out of 101 showed presence of SARS-CoV-2 RNA. No VeroE6 cell infection was detected with viral isolates, thus suggesting no presence of SARS-CoV-2 virions in corneal specimens and storage media. CONCLUSIONS: The presence of SARS-CoV-2 in cornea specimens would seem to be more likely due to prolonged detection of RNA rather than to active viral replication, with very low risk of infectivity and transmission through keratoplasty.

5†The impact of the pandemic on ocular tissue procurement : the experience of fondazione banca degli Occhi del veneto.

INTRODUCTION: Since 21 February 2020, the day that the first Italian COVID-19 case was identified, the organizational and regulatory conditions for ocular tissue donation have undergone numerous changes in order to guarantee safety and quality. Herewith we report the key responses of the procurement programme to these challenges. MATERIALS AND METHODS: A retrospective analysis of the ocular tissue procured between 1 January 2020 and 30 September 2021 is reported. RESULTS: 9224 ocular tissues were procured during the study period (weekly average: 100 ± 21 tissues, mean ± SD; down to 97 ± 24, if only 2020 is considered). During the first wave, the weekly average reached 80 ± 24 tissues, a significant reduction if compared to the first 8 weeks of the year (124 ± 22 tissues/week, p<0.001), falling to 67 ± 15 tissues/week during the lock-down period. Considering the ocular tissues collected in the Veneto Region alone, the weekly mean was 68 ± 20, a reduction when compared to the first 8 weeks of the year (102 ± 23, p<0.001), arriving at 58 ± 15 tissues/week during the lock-down period. The percentage of healthcare professionals who tested positive during the first wave was on average 12% of the positive cases in the whole country, and equal to 18% in the Veneto Region alone. During the second wave, the mean weekly recovery of ocular tissue was 91 ± 15 and 77 ± 15 in the Veneto Region, compared to positive cases of healthcare professionals of 4% across Italy and in the Veneto Region. During the third wave, the overall weekly mean recovery rate was 107 ± 14, and 87 ± 13 in the Veneto Region, with only 1% of positive cases among healthcare professionals in Italy and in the Veneto Region. CONCLUSIONS: The most dramatic decrease of ocular tissue recovery occurred during the first wave of COVID-19, notwithstanding the lower number of infected people. This phenomenon can be attributed to different factors: a high percentage of positive cases and/or contacts among potential donors; the number of infections among healthcare professionals, favoured by the lack of personal protection equipement and the still partial knowledge of the disease; the exclusion of donors with bilateral pneumonia. Subsequently, the system was better organized with the assimilation of new knowledge about the virus, overcoming the initial fears about transmission and thus guaranteeing the resumption and maintenance of donations.

The role of the comprehensive complication index for the prediction of survival after liver transplantation.

In the last years, several scoring systems based on pre- and post-transplant parameters have been developed to predict early post-LT graft function. However, some of them showed poor diagnostic abilities. This study aims to evaluate the role of the comprehensive complication index (CCI) as a useful scoring system for accurately predicting 90-day and 1-year graft loss after liver transplantation. A training set (n = 1262) and a validation set (n = 520) were obtained. The study was registered at https://www.ClinicalTrials.gov (ID: NCT03723317). CCI exhibited the best diagnostic performance for 90 days in the training (AUC = 0.94; p < 0.001) and Validation Sets (AUC = 0.77; p < 0.001) when compared to the BAR, D-MELD, MELD, and EAD scores. The cut-off value of 47.3 (third quartile) showed a diagnostic odds ratio of 48.3 and 7.0 in the two sets, respectively. As for 1-year graft loss, CCI showed good performances in the training (AUC = 0.88; p < 0.001) and validation sets (AUC = 0.75; p < 0.001). The threshold of 47.3 showed a diagnostic odds ratio of 21.0 and 5.4 in the two sets, respectively. All the other tested scores always showed AUCs < 0.70 in both the sets. CCI showed a good stratification ability in terms of graft loss rates in both the sets (log-rank p < 0.001). In the patients exceeding the CCI ninth decile, 1-year graft survival rates were only 0.7% and 23.1% in training and validation sets, respectively. CCI shows a very good diagnostic power for 90-day and 1-year graft loss in different sets of patients, indicating better accuracy with respect to other pre- and post-LT scores.Clinical Trial Notification: NCT03723317.

ALPPS for Locally Advanced Intrahepatic Cholangiocarcinoma: Did Aggressive Surgery Lead to the Oncological Benefit? An International Multi-center Study.

BACKGROUND: ALPPS is found to increase the resectability of primary and secondary liver malignancy at the advanced stage. The aim of the study was to verify the surgical and oncological outcome of ALPPS for intrahepatic cholangiocarcinoma (ICC). METHODS: The study cohort was based on the ALPPS registry with patients from 31 international centers between August 2009 and January 2018. Propensity score matched patients receiving chemotherapy only were selected from the SEER database as controls for the survival analysis. RESULTS: One hundred and two patients undergoing ALPPS were recruited, 99 completed the second stage with median inter-stage duration of 11 days. The median kinetic growth rate was 23 ml/day. R0 resection was achieved in 87 (85%). Initially high rates of morbidity and mortality decreased steadily to a 29% severe complication rate and 7% 90-day morbidity in the last 2 years. Post-hepatectomy liver failure remained the main cause of 90-day mortality. Multivariate analysis revealed insufficient future liver remnant at the stage-2 operation (FLR2) to be the only risk factor for severe complications (OR 2.91, p = 0.02). The propensity score matching analysis showed a superior overall survival in the ALPPS group compared to palliative chemotherapy (median overall survival: 26.4 months vs 14 months; 1-, 2-, and 3-year survival rates: 82.4%, 70.5% and 39.6% vs 51.2%, 21.4% and 11.3%, respectively, p < 0.01). The survival benefit, however, was not confirmed in the subgroup analysis for patients with insufficient FLR2 or multifocal ICC. CONCLUSION: ALPPS showed high efficacy in achieving R0 resections in locally advanced ICC. To get the most oncological benefit from this aggressive surgery, ALPPS would be restricted to patients with single lesions and sufficient FLR2.

Videolaparoscopic microwave ablation in patients with HCC at a European high-volume center: Results of 815 procedures.

BACKGROUND: Videolaparoscopic (VL) microwave ablation (MWA) is not included in most of the international guidelines as a therapeutic option for hepatocellular carcinoma (HCC). Aim of this study was to assess the safety of VL MWA in patients with HCC for whom resection or percutaneous ablation is unsuitable. METHODS: A retrospective analysis was performed on a prospective database of patients with HCC treated with VL MWA at our institution from 2009 to 2016. Patient demographics, operational characteristics, and complications were recorded. Statistical analysis was performed to identify safety profile, overall survival and recurrence rate. RESULTS: A total of 815 VL MWA were performed in 674 patients with a mean age of 64 years. Patients had a mean Model for End-stage Liver Disease score of 10 (±3); 32.8% were Child B, 44.1% Barcelona Clinic Liver Cancer B-C. Perioperative mortality was 0.4%. Overall morbidity was 30.8%, with Dindo-Clavien complications ≥3 in 2%. The median length of stay was 2 days. In 43.1% VL MWA was the first-line therapy. Overall 1-, 3-, and 5-year survival rates were 81.9%, 54.9%, and 35.9%. CONCLUSIONS: The present is the largest series of VL ablation and the bigger number of patients with HCC treated with MW reported nowadays. It confirms the safety of a minimally invasive procedure for patients with HCC when resection or percutaneous ablation is not feasible.