The aim of this work is to quantify the total protein and growth factors content in a tissue-suspension obtained from processed human amniotic membrane (hAM). hAM was collected, frozen, freeze dried, powdered and sterilized by γ-irradiation. At each step of the process, samples were characterized for the total protein amounts by a Bradford protein assay and for the growth factor concentrations by ELISA test of the tissue suspensions. Frozen-hAM samples show higher release of total proteins and specific growth factors in the tissue suspension in comparison with freeze-dried hAM. We observed that even if the protein extraction is hindered once the tissue is dried, the powdering process allows a greater release in the tissue suspension of total proteins and growth factors after tissue re-solubilization in comparison with only the freeze-drying process (+91 ± 13% for EGF, +16 ± 4% for HGF, +11 ± 5% for FGF, +16 ± 9% for TGF-β1), and a greater release of EGF (85 ± 10%) in comparison with only the freezing process, because proteins become much readily solubilized in the solution. According with these results, we describe a protocol to obtain a new sterile biological product from hAM tissue, with well-known effects of thermal, mechanical and physical processes on the total protein and grow factors contents.
To investigate the impact of Coronavirus Disease-2019 lockdown on the Italian Eye Bank organization. In this national retrospective, multicentric, cohort study, data from the Italian Eye Bank during both the lockdown and the first month after the lockdown period were retrieved. We compared the Italian Eye Bank metrics with the same timeframe of 2019 and 2018. Data from 13 out of 13 (100%) Italian Eye Banks were included in the analysis. A statistically significant reduction in the number of donor corneas retrieved in 2020 was found as compared to the same period in 2019 and in 2018, respectively (2020 = 1284; 2019 = 3088; 2018 = 3221; ANOVA: p < 0.0001). Only 534 corneas have been distributed by Eye Banks during the COVID-19-lockdown period (2020 = 534; 2019 = 1220; 2018 = 1237. ANOVA: p < 0.0001). Similarly, the number of wasted corneas due to postponed or cancelled surgeries was 421, resulting in a considerable increase as compared to the previous 2 years (2020 = 421; 2019 = 67; 2018 = 84; ANOVA: p = 0.0035). Overall, 45 donor corneas were rejected in accordance with the guidance of the Italian National Health Institute Italian National Transplant Centre (CNT). SARS-CoV-2 pandemic has profoundly affected every social and medical field, including the Eye Bank procurement and distribution programs. The current data collected from all the Italian Eye Banks highlights the present and the forthcoming difficulties that the Eye Bank community is going to experience, as for the ongoing pandemic.
This new therapy, which is less traumatic than an implant of amniotic membrane, is safe, and can be repeated for a long period, could help patients with corneal superficial defects.
Purpose: This study aims to explore and characterize healthy eye microbiota. Methods: Healthy subjects older than 18 years were selected for this descriptive cross-sectional study. Samples were collected with an eSwab with 1 mL of Liquid Amies Medium (Copan Brescia, Italy). Following DNA extraction, libraries preparation, and amplification, PCR products were purified and end-repaired for barcode ligation. Libraries were pooled to a final concentration of 26 pM. Template preparation was performed with Ion Chef according to Ion 510, Ion 520, and Ion 530 Kit-Chef protocol. Sequencing of the amplicon libraries was carried out on a 520 or 530 chip using the Ion Torrent S5 system (Thermo Fisher; Waltham, MA, USA). Raw reads were analyzed with GAIA (v 2.02). Results: Healthy eye microbiota is a low-diversity microbiome. The vast majority of the 137 analyzed samples were highly enriched with Staphylococcus, whereas only in a few of them, other genera such as Bacillus, Pseudomonas, and Corynebacterium predominate. We found an average of 88 genera with an average Shannon index of 0.65. Conclusion: We identified nine different ECSTs. A better understanding of healthy eye microbiota has the potential to improve disease diagnosis and personalized regimens to promote health.
The results showed that the method of preservation and distribution of dehydrated, 4 °C stored, and rehydrated lenticules is safe, cheap, and simple. Furthermore, it does not appear to influence the outcomes of the DALK surgery.
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