Publications by authors named "Hossein Elbadawy"

20 Publications

  • Page 1 of 1

Design of molecular hybrids of phthalimide-triazole agents with potent selective MCF-7/HepG2 cytotoxicity: Synthesis, EGFR inhibitory effect, and metabolic stability.

Bioorg Chem 2021 Jun 22;111:104835. Epub 2021 Mar 22.

Pharmacognosy and Pharmaceutical Chemistry Department, College of Pharmacy, Taibah University, Al-Madinah Al-Munawarah, Saudi Arabia; Pharmaceutical Organic Chemistry Department, Faculty of Pharmacy, Al-Azhar University, 11884 Nasr City, Cairo, Egypt.

This study reports an efficient and convenient click chemistry synthesis of a novel series of phthalimide scaffold linked to 1,2,3 triazole ring and terminal lipophilic fragments. Structures of newly synthesized compounds were well characterized by different spectroscopic tools. In vitro MTT cytotoxicity assay was performed comparing the cytotoxic effects of newly synthesized compounds to staurosporine using three different types: human liver cancer cell line (HepG2), Michigan cancer foundation-7 (MCF-7) and human colorectal carcinoma cell line (HCT116). The initial screening showed excellent to moderate anticancer activity for these newly synthesized compounds with high degree of cell line selectivity with micromolar (µM) half maximal inhibitory concentration (IC) values against tumor cells. The SAR analysis of these derivatives confirmed the role of molecular fragments including phthalimide, linker, triazole, and terminal tails in correlation to activity. In addition, enzymatic inhibitory assay against wild type EGFR was performed for the most active compounds to get more details about their mechanism of action. In order to further explore their binding affinities, molecular docking simulation was studied against EGFR site. The results obtained from molecular docking study and those obtained from cytotoxic screening were correlated. One of the most prominent analogs is (6f) with terminal disubstituted ring and amide linker showed selective MCF-7 cytotoxicity profile with IC 0.22 µM and 79 nM to EGFR target. Extensive structure activity relationship (SAR) analyses were also carried out. The pharmacokinetic profile of (6f) was studied showing good metabolic stability and long duration behavior. This design offered a potent selective anticancer phthalimide-triazole leads for further optimization in cancer drug discovery.
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http://dx.doi.org/10.1016/j.bioorg.2021.104835DOI Listing
June 2021

Prevalence of dry eye syndrome in association with the use of contact lenses in Saudi Arabia.

BMC Ophthalmol 2021 Mar 23;21(1):147. Epub 2021 Mar 23.

Department of Pharmacology and Toxicology, College of Pharmacy, Taibah university, Madinah, Kingdom of Saudi Arabia.

Background: Dry eye disease is a tear film disorder which can cause discomfort to patients and negatively affect vision acuity. A number of risk factors has been reported to affect the incidence and severity of dry eye syndrome (DES). The aim is to study the prevalence of DES in Saudi Arabia and the factors affecting the severity of DES in relation to the use of contact lenses.

Methods: A cross-sectional questionnaire-based study was conducted on 310 participants using the ocular surface disease index (OSDI) questionnaire and the eye dryness part from contact lens questionnaire-8 (CLDEQ-8). Dry eye OSDI scores were compared across different epidemiological and risk factors with focus on the use of contact lenses. Pearson and Spearman's correlation coefficients were used to analyze the frequency of contact lenses usage in relation to OSDI scores. Student's t-test and one-way analysis of variance (ANOVA) tests were used to compare means of two or more than two groups, respectively.

Results: Forty eight (15.5%) of participants did not have any degree of DES, achieving an OSDI score between 0 and 12. Forty participants (12.9%) scored from 13 to 22, (mild DES), 44 (14.2%) were moderate, scoring 23-32 on the OSDI, while those who scored above 33 were 178 (57.4%) had severe DES. The mean score for all participants was 37.8. A high percentage of participants (84.5%) had some degree of DES. There was a strong positive correlation between OSDI score and the frequency of the feeling of dry eye and a moderate positive correlation between OSDI score and the intensity of dryness feeling. Out of 310 participants, 136 (43.9%) indicated using contact lenses. There was no significant association between the use of contact lenses per se and DES, however, those who used contact lenses more frequently had significantly higher OSDI scores.

Conclusions: Dry eye syndrome is a widespread, underdiagnosed condition in Saudi Arabia. The frequency of contact lenses use may contribute to the incidence of DES.
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http://dx.doi.org/10.1186/s12886-021-01912-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7986502PMC
March 2021

The detection of SARS-CoV-2 in outpatient clinics and public facilities during the COVID-19 pandemic.

J Med Virol 2021 05 10;93(5):2955-2961. Epub 2021 Feb 10.

Department of Medical Laboratory Technology, College of Applied Medical Sciences, Taibah University, Madinah, Kingdom of Saudi Arabia.

The transmission of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can occur through an airborne route, in addition to contaminated surfaces and objects. In hospitals, it has been confirmed by several studies that SARS-CoV-2 can contaminate surfaces and medical equipment especially in hospitals dedicated to coronavirus disease 2019 (COVID-19) patients. The aim of this study was to detect the contamination of hands, objects, and surfaces in isolation rooms and also in outpatients' clinics in hospitals and polyclinics. Environmental contamination of public high-touch surfaces in public facilities was also investigated during an active COVID-19 pandemic. Random swabs were also taken from public shops, pharmacies, bakeries, groceries, banknotes, and automated teller machines (ATMs). Samples were analyzed for SARS-CoV-2 positivity using real-time polymerase chain reaction. In the COVID-19 regional reference hospital, only 3 out of 20 samples were positive for SARS-CoV-2 RNA. Hand swabs from SARS-CoV-2-positive patients in isolation rooms were occasionally positive for viral RNA. In outpatients' clinics, door handles were the most contaminated surfaces. Dental chairs, sinks, keyboards, ophthalmoscopes, and laboratory equipment were also contaminated. Although no positive swabs were found in shops and public facilities, random ATM swabs returned a positive result for SARS-CoV-2. Although there is no longer a focus on COVID-19 wards and isolation hospitals, more attention is required to decontaminate frequently touched surfaces in health-care facilities used by patients not diagnosed with COVID-19. Additionally, high-touch public surfaces such as ATMs require further disinfection procedures to limit the transmission of the infection.
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http://dx.doi.org/10.1002/jmv.26819DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8014553PMC
May 2021

Enhancement of atorvastatin oral bioavailability via encapsulation in polymeric nanoparticles.

Int J Pharm 2021 Jan 24;592:120077. Epub 2020 Nov 24.

Department of Pharmaceutics, Faculty of Pharmacy, King Khalid University, Abha, Saudi Arabia.

Despite the fact that atrovastatin (At) is being one of the bestselling statins used to prevent complicated cardiovascular diseases, its low oral bioavailability decreases its clinical relevance. Herein, incorporation of At into ethylcellulose nanoparticles (At-NPs) was executed to test if it would enhance its oral bioavailability. The emulsification-evaporation method was used to prepare the At-NPs. The prepared nanoparticles were characterized by measuring the particle size, zeta potential as well as using FTIR, DSC, and XRD examination. The entrapment efficiency, drug content, and the in vitro release behavior of At-NPs were also examined. The in vivo oral bioavailability of the selected At-NPs formula was tested after being given orally to New Zealand rabbits. The nanoparticles obtained had a high drug content and a distinct spherical shape but with varying sizes. No physical or chemical interactions were detected between At and the nanoparticles as confirmed by FTIR, DSC, and XRD. The in vitro release study of At from the prepared At-NPs has shown nanoparticles size-dependent release behavior. The in vivo oral absorption testing confirmed the bioavailability of the prepared At-NPs to be as follows: (C = 940 ng/ml and AUC = 8759 ng.h/ml) > Lipitor® (C = 635 ng/ml and AUC = 4367 ng.h/ml) > At (C = 515 ng/ml and AUC = 2517 ng.h/ml). These results revealed that the oral formula of At-NPs increases the bioavailability of At 3.87 times. This makes ethylcellulose nanoparticles an esteemed candidate nano-vehicle for At, increasing its bioavailability and thus improving its clinical relevance.
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http://dx.doi.org/10.1016/j.ijpharm.2020.120077DOI Listing
January 2021

Generation of Combinatorial Lentiviral Vectors Expressing Multiple Anti-Hepatitis C Virus shRNAs and Their Validation on a Novel HCV Replicon Double Reporter Cell Line.

Viruses 2020 09 18;12(9). Epub 2020 Sep 18.

Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.

Despite the introduction of directly acting antivirals (DAAs), for the treatment of hepatitis C virus (HCV) infection, their cost, patient compliance, and viral resistance are still important issues to be considered. Here, we describe the generation of a novel JFH1-based HCV subgenomic replicon double reporter cell line suitable for testing different antiviral drugs and therapeutic interventions. This cells line allowed a rapid and accurate quantification of cell growth/viability and HCV RNA replication, thus discriminating specific from unspecific antiviral effects caused by DAAs or cytotoxic compounds, respectively. By correlating cell number and virus replication, we could confirm the inhibitory effect on the latter of cell over confluency and characterize an array of lentiviral vectors expressing single, double, or triple cassettes containing different combinations of short hairpin (sh)RNAs, targeting both highly conserved viral genome sequences and cellular factors crucial for HCV replication. While all vectors were effective in reducing HCV replication, the ones targeting viral sequences displayed a stronger antiviral effect, without significant cytopathic effects. Such combinatorial platforms as well as the developed double reporter cell line might find application both in setting-up anti-HCV gene therapy approaches and in studies aimed at further dissecting the viral biology/pathogenesis of infection.
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http://dx.doi.org/10.3390/v12091044DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7551853PMC
September 2020

The metabolic disorders associated with chronic consumption of soft and energy drinks in rats.

Acta Biochim Pol 2020 Mar;67(1):79-84

1Department of Pharmacology and Toxicology (subdivision of Biochemistry), College of Pharmacy, Taibah University, Medina, Kingdom of Saudi Arabia; 2Department of Biochemistry, Faculty of Pharmacy, Minia University, Minia, Egypt.

Background: Energy Drinks (EDs) and Soft Drinks (SDs) are widely consumed among adolescents and young adults. These drinks contain variable amounts of caffeine which is a central nervous system stimulator; in addition to sugar, taurine, vitamins and herbal extracts. Several adverse effects have been reported for the excessive consumption of caffeine and sugar.

Aim: This work aimed at providing a comparison between the effect of chronic consumption of both drinks on metabolism biochemically as well as at the histopathological level.

Methods: Adult albino rats were randomly divided into three groups and treated for 4 weeks. Animals received water (control, group 1), 12.5 ml/kg/day of either Pepsi® (SD, group 2) or Power Horse® (ED, group 3). All animals had free access to water and standard animal chow.

Results: ED and SD groups showed a significant weight gain compared to control. ED animals showed a significant increase in serum urea, hyperlipidemia and hyperglycemia in comparison to control and SD groups. Serum uric acid significantly increased in ED and SD groups. ED group showed congestion and inflammation in their renal tissues in addition to splenomegaly and increased phagocyte infiltration.

Conclusion: The high caffeine-sugar content in ED exerts a more significant influence on the metabolic pathways than SDs. Both increase the incidence of cardiovascular diseases and tissue inflammation due to their effect on lipid profile and blood glucose. The other ingredients in EDs may play a role in the observed metabolic disturbances. Chronic use of EDs should be especially discouraged to avoid these negative effects.
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http://dx.doi.org/10.18388/abp.2020_2914DOI Listing
March 2020

Protective effects of Ajwa date extract against tissue damage induced by acute diclofenac toxicity.

J Taibah Univ Med Sci 2019 Dec 27;14(6):553-559. Epub 2019 Nov 27.

Department of Pharmacognosy and Pharmaceutical Chemistry, College of Pharmacy, Taibah University, Almadinah Almunawwarah, KSA.

Objectives: To investigate the tissue-protective effects of Ajwa date fruits (a Prophetic medicinal remedy) against acute diclofenac toxicity.

Methods: Albino Sprague-Dawley rats were allocated to four experimental groups: a negative control group, an Ajwa-only group that received 2 g/kg of Ajwa date extract (ADE) orally, an acute diclofenac toxicity group that received 200 mg diclofenac once intraperitoneally, and a treatment group that received diclofenac and ADE after 4 h. Histological examinations of rat lung and liver tissues were performed.

Results: Acute diclofenac toxicity caused marked hepatic derangements, such as congested central veins, congested blood sinusoids, hyaline degeneration, and hepatocyte necrosis. Toxic diclofenac overdose resulted in markedly congested alveolar capillaries and alveolar haemorrhages, thick edematous alveolar walls, and edema fluid exudates in the alveoli. Upon treatment with ADE, significant reduction in diclofenac-induced hepatic and pulmonary derangements were observed.

Conclusion: ADE is a safe, tissue-protective nutritional agent that alleviates cellular and tissue-damaging effects due to acute diclofenac toxicity. ADE relieved hepatic and pulmonary changes induced by acute diclofenac toxicity. The use of ADE is recommended for the treatment of acute diclofenac toxicity.
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http://dx.doi.org/10.1016/j.jtumed.2019.10.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6940670PMC
December 2019

Introducing of acyclonucleoside analogues tethered 1,2,4-triazole as anticancer agents with dual epidermal growth factor receptor kinase and microtubule inhibitors.

Bioorg Chem 2020 01 23;94:103446. Epub 2019 Nov 23.

Department of Chemistry, Faculty of Sciences, Taibah University, Al-Madinah Al-Munawarah 30002, Saudi Arabia; Laboratoire de Chimie & Electrochimie des Complexes Métalliques (LCECM) USTO-MB, Department of Chemistry, Faculty of Sciences, University of Sciences and Technology Mohamed Boudiaf, B.p. 1505 El M nouar, Oran 31000, Algeria.

This study reports an efficient and convenient regioselective synthesis of a novel series of S- and S,N-bis(acyclonucleoside) analogues carrying 5-(2-chlorophenyl)-2,4-dihydro-1,2,4-triazole-3-thione. A facile and straightforward synthesis of thiazolotriazole and triazolothiazines has also been reported. Structures of all newly synthesized compounds were well characterized by infrared IR, H and C nuclear magnetic resonance (NMR) and mass (MS) spectra analyses. Cytotoxic screening was performed according to (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium (MTT) assay method using staurosporine as a reference drug against three different types: human liver cancer cell line (Hep G2), Michigan cancer foundation-7 (MCF-7) and human colorectal carcinoma cell line (HCT116). These data showed considerable anticancer activity for these newly synthesized compounds. Biological data for most of the S-acyclonucleoside analogues and S,N-bis(acyclonucleoside) analogues showed excellent activity with micromolar (µM) half maximal inhibitory concentration (IC) values against tumor cells. EGFR assay and tubulin inhibition assay analysis were performed for the most active compounds to get more details about their mechanism of action. In order to assess and explain their binding affinities, molecular docking simulation was studied against EGFR and tubulin binding sites. The results obtained from molecular docking study and those obtained from cytotoxic screening were correlated. Extensive structure activity relationship (SAR) analyses were also carried out.
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http://dx.doi.org/10.1016/j.bioorg.2019.103446DOI Listing
January 2020

Improved solubility, dissolution, and oral bioavailability for atorvastatin-Pluronic® solid dispersions.

Int J Pharm 2020 Jan 28;574:118891. Epub 2019 Nov 28.

Department of Pharmaceutics, Faculty of Pharmacy, King Khalid University, Abha, Saudi Arabia.

Despite the status of atorvastatin (AT) as one of the top selling statins for prophylaxis against primary and secondary cardiovascular diseases, its limited oral absorption limits its full therapeutic benefits. Herein, formulations of AT with amphiphilic carriers (Pluronic F127® and Pluronic F68®) were developed in the form of hard capsules to improve in vitro solubility and dissolution, as well as in vivo oral bioavailability. Prepared formulas were characterized by assessing solubility improvements in the carrier solution and examining the FTIR, DSC, and X-RPD profiles for each formula. The dissolution rate and absorption were also examined after oral administration to New Zealand rabbits. The solubility of AT was improved by the incorporation of either Pluronic F127® or Pluronic F68®. No chemical changes or interactions were detected using X-RPD, DSC, and FTIR characterization. Dissolution profiles revealed an increase in the rate and maximum amount of dissolved AT and showed that up to 93% of the AT content was dissolved within 30 min. In vivo absorption of the tested formula (C = 1146 ng/ml and AUC0-12 to 9,993.4 ng.h/ml) was greater than Lipitor® (C = 642.3 ng/ml and AUC0-12 = 4427.4 ng.h/ml) and AT (C = 517.6 ng/ml and AUC0- 12 = 2,473.7 ng.h/ml). In conclusion, the formulation of AT with Pluronics® profoundly augments the dissolution behavior and absorption of AT and may serve as a useful approach for improving AT therapeutic and clinical efficacy.
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http://dx.doi.org/10.1016/j.ijpharm.2019.118891DOI Listing
January 2020

Recombinant human serum albumin for corneal preservation.

Acta Ophthalmol 2018 Feb 21;96(1):e79-e86. Epub 2017 Jun 21.

International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy.

Purpose: To study the performance of a completely synthetic organ culture (OC) preservation system containing recombinant human serum albumin (rHSA) for preservation of human donor corneas.

Methods: Twenty-four paired donor corneas were randomly collected, and one cornea from each donor was preserved in synthetic (experimental) and serum-based media (control). The tissues were assessed at day 0; after 6 days of preservation at room temperature (RT) in Cornea Trans and Cornea Prep II ; after 28 days at 31°C in Cornea Syn [with rHSA] and Cornea Max [with foetal calf serum (FCS)] and; 4-day post deswelling in Cornea Trans and Cornea Jet . Thickness was determined with optical coherence tomography (OCT) and transparency with a validated, custom device. Morphology, endothelial cell density (ECD) and mortality were observed after treating the tissues with Trypan blue and sucrose. Glucose uptake by the cells was analysed. Data were compared using non-parametric paired Wilcoxon tests with p < 0.05 deemed significant. Histology using periodic acid-schiff (PAS), expressions of p63, CK12, αSMA and ZO-1 were analysed, and cell apoptosis postpreservation was studied.

Results: Corneas stored in synthetic media showed a higher and statistically significant value as compared to serum-based media in terms of viable endothelial cell density (VECD), mortality, morphology and glucose uptake postpreservation. Histology showed presence of all the layers, all the markers were expressed, and no apoptosis was observed in either media.

Conclusion: The new synthetic preservation system containing rHSA (and other confidential constituents) showed better preservation effects than traditional media containing serum.
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http://dx.doi.org/10.1111/aos.13498DOI Listing
February 2018

Towards xeno-free cultures of human limbal stem cells for ocular surface reconstruction.

Cell Tissue Bank 2017 Dec 26;18(4):461-474. Epub 2017 May 26.

Fondazione Banca degli Occhi del Veneto, c/o Padiglione G. Rama - Via Paccagnella 11, 30174, Zelarino, Venice, Italy.

Isolated limbal epithelial stem cells (LESCs) were cultured with or without a 3T3 murine fibroblast feeder-layer (FL) in 4 different culture media on culture plates or on denuded human amniotic membrane (AM) support and fibrin gel support: (1) control medium supplemented with fetal bovine serum; (2) control medium supplemented with the synthetic serum "XerumFree™ XF205" (XF); (3) CnT-20 medium supplemented with "XerumFree™ XF205" (CnT-XF) and (4) CnT-20 medium supplemented with human AB serum (CnT-AB). The three xenogeneic media were compared to standard condition (control + FL) and parameters assessed included cell morphology, proliferative potential, number of passages, assessment of clonogenic and abortive colonies, life span, ∆Np63α expression and epithelial morphology on AM. During serial cultivation of LESCs, most of the tested xeno-free media supported similar numbers of cell passages, total colony number, cumulative cell doublings (CCD) rates and expression of ∆Np63α compared to control. The conditions cultivated with a FL showed a non-statistically significant higher number of cell passages and CCD rates before senescence when compared to the same conditions cultured without FL. Except for the control medium, only XF medium enabled the growth of cells on AM. The expression of ∆Np63α was comparable in all the cultures grown onto AM, when compared to the controls on fibrin gel. In conclusion, the xeno-free media enabled LESC culture both on plastic and on denuded human AM. Despite the analyses were carried out in a statistically low number of samples and need re-assessment in a larger cohort, our results suggest that the production of a completely xeno-free LESC graft could be beneficial for future clinical applications.
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http://dx.doi.org/10.1007/s10561-017-9632-7DOI Listing
December 2017

Role of dextran in maintaining adhesive and stiffness properties of prestripped DMEK lenticules.

Eur J Ophthalmol 2017 May 25;27(3):270-277. Epub 2016 Nov 25.

International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice - Italy.

Purpose: To investigate the adhesive and stiffness properties of prestripped Descemet membrane endothelial keratoplasty (DMEK) lenticules in different preservation conditions (with and without dextran).

Methods: The study included 3 conditions: (C1) tissues collected from tissue culture media (TCM), stripped and preserved in TCM; (C2) tissues collected from transport media (TM) (TCM supplemented with 6% dextran T-500), stripped and preserved in TM; and (C3) tissues collected from TCM, stripped and preserved in TM. Using a hinge, 9.5-mm stripped DMEK lenticules were restored back on the stroma and preserved for 4 days at room temperature (RT) in different conditions as above. Nine tissues, 3 from each condition, were used to check the adhesive (fibronectin, laminin, and vitronectin) and elastic properties (fibrillin, elastin, and collagen VI) using different antibodies. Six tissues, 2 from each condition, were used to check the stiffness properties after preservation using atomic force microscopy (AFM) nanoindentation method.

Results: On the Descemet membrane, fibronectin was strongly expressed in C2 and C3, whereas laminin was intense in C2 postpreservation. Vitronectin was expressed in all the conditions. Elastic proteins were not expressed in either of the conditions apart from collagen VI, which was expressed on the posterior stroma. Atomic force microscopy showed higher stiffness in C3 and an insignificant but lower rigidity in C2 as compared to C1.

Conclusions: The tissues from C2 showed expression of adherent proteins and lower stiffness. Dextran may be suitable in preservation of DMEK grafts before and after preparation. Less stiff tissues may help reduce manipulations required in the recipient eye during DMEK surgery.
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http://dx.doi.org/10.5301/ejo.5000906DOI Listing
May 2017

Corneal Epithelial Stem Cells Repopulate the Donor Area within 1 Year from Limbus Removal for Limbal Autograft.

Ophthalmology 2016 12 21;123(12):2481-2488. Epub 2016 Sep 21.

Research Centre, Fondazione Banca degli Occhi del Veneto, Venice, Italy; Department of Pharmacology and Toxicology, College of Pharmacy, Taibah University, Al-Madinah Al-Munawarah, Kingdom of Saudi Arabia.

Purpose: To determine whether limbal epithelial stem cells (LESCs) repopulate the site harvested for limbal autograft transplantation (LAT), the expression of LESCs markers was evaluated in bioptic specimens obtained from the donor area 12 months or more after surgery.

Design: Interventional case series.

Participants: Patients who underwent LAT for unilateral acquired limbal stem cell deficiency after chemical burn.

Methods: Corneal limbal explants were obtained from 2 sites, the harvested area and the untouched control area, in the donor eyes of 6 patients who previously underwent LAT for unilateral acquired limbal stem cell deficiency after chemical burn. Limbal epithelial stem cells were isolated, and cellular, immunohistochemistry, and histologic parameters were assessed to compare differences between LESCs isolated from harvested or control sites.

Main Outcome Measures: Presence of LESCs 1 year or more after LAT.

Results: Specific markers (p63, Ki67, K12), percentage of LESCs, cell doubling, and number of passages in culture did not differ significantly between harvested and control sites. However, the distinctive structure of the palisades of Vogt was found only in 2 of 6 harvested sites.

Conclusions: Limbal epithelial stem cells repopulate the donor site as early as 1 year after limbus removal for LAT. Autologous transplantation of conjunctiva and limbus are safe procedures and can be performed in cases that cannot be treated by simple grafting of LESCs cultured ex vivo.
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http://dx.doi.org/10.1016/j.ophtha.2016.08.018DOI Listing
December 2016

Effect of connexin 43 inhibition by the mimetic peptide Gap27 on corneal wound healing, inflammation and neovascularization.

Br J Pharmacol 2016 10 26;173(19):2880-93. Epub 2016 Aug 26.

International Center for Ocular Physiopathology, The Veneto Eye Bank Foundation, Venice, Italy.

Background And Purpose: The connexin 43 (Cx43) mimetic peptide Gap27 was designed to transiently block the function of this gap junction. This study was undertaken to investigate the effect of Gap27 on corneal healing, inflammation and neovascularization.

Experimental Approach: The effect of Gap27 on wound healing, inflammation and vascularization was assessed in primary human corneal epithelial cells (HCEC) in vitro and whole human corneas ex vivo, and in an in vivo rat wound healing model.

Key Results: Gap27 enhanced the wound closure of HCEC in vitro and accelerated wound closure and stratification of epithelium in human corneas ex vivo, but did not suppress the corneal release of inflammatory mediators IL-6 or TNF-α in vivo. In human corneas ex vivo, F4/80 positive macrophages were observed around the wound site. In vivo, topical Gap27 treatment enhanced the speed and density of early granulocyte infiltration into rat corneas. After 7 days, the expressions of TNF-α and TGFβ1 were elevated and correlated with inflammatory cell accumulation in the tissue. Additionally, Gap27 did not suppress VEGF release in organotypic culture, nor did it suppress early or late VEGFA expression or neovascularization in vivo.

Conclusions And Implications: Gap27 can be effective in promoting the healing of superficial epithelial wounds, but in deep stromal wounds it has the potential to promote inflammatory cell migration and accumulation in the tissue and does not suppress the subsequent neovascularization response. These results support the proposal that Gap27 acts as a healing agent in the transient, early stages of corneal epithelial wounding.
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http://dx.doi.org/10.1111/bph.13568DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5055138PMC
October 2016

Polarization of human donor corneas.

Cell Tissue Bank 2016 Jun 27;17(2):233-9. Epub 2016 Feb 27.

Phronema srl, via Junipero Serra, 19, 70125, Bari, Italy.

To investigate the de-orientation effect of DSAEK grafts by observing the cross patterns and polarization power of human donor corneas using a polarizing device (Lumaxis(®)). Forty human donor corneas were placed in small petri-plates with epithelial side facing up. Polarizing power (arbitrary unit) and crosses were monitored and recorded by the software. The tissue was marked at 'Superior' position to ensure that the base and the polarizer are in alignment with each other after the cut. The anterior lamellar cut was performed using microkeratome. The lenticule was placed back in the same position as marked to mimic the alignment. The tissue was further rotated by 45° ensuring that the base of the cornea and the polarizer were in alignment. The polarization power and 'crosses' were identified at each step. The average of forty corneas from pre-cut to post-45° angular change showed statistically significant difference (p < 0.05) in terms of polarizing power. The cross-shaped pattern deformed and lost the sharpness towards 45° angle. However, multiple variances in terms of 'cross-patterns' were observed throughout the study. Lumaxis(®) was able to determine the worst quality tissue in terms of polarization (no black zone and crosses). Despite the quality of cross pattern which can be used as an additional objective parameter to evaluate the optical properties of the corneal tissue, this preliminary study needs to be further justified in terms of clinical relevance whether polarization changes with oriented or de-oriented grafts have any effects and consequences on the visual acuity.
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http://dx.doi.org/10.1007/s10561-016-9546-9DOI Listing
June 2016

Transplantation failure due to inadvertent reversal of human donor corneas.

Eur J Ophthalmol 2016 Feb 15;26(2):e21-3. Epub 2016 Feb 15.

International Center for Ocular Physiopathology (ICOP), Fondazione Banca Degli Occhi Del Veneto Onlus, Venice - Italy.

Purpose: To emphasize and create awareness of the possibility of inadvertently grafting an inverted corneal button and describe 4 cases that showed this phenomenon on human donor corneas during preservation and transportation from an eye bank.

Methods: Three out of 4 tissues showed inadvertent inversion during transportation and 1 was identified as inverted during preservation in the eye bank. Out of the 3 tissues that were shipped, 2 tissues were transplanted and 1 was identified as inverted prior to transplantation and shipped back to the eye bank.

Results: Four tissues showed inversion at different time points. The anatomy was clearly visible with a naked eye that showed the presence of trabecular meshwork on its outside along with some residual choroid. The endothelial cell count was in the range of transplantation without any mortality even after inversion. The inverted corneas showed a mountain-like shape with grooves as compared to the normal cornea.

Conclusions: Previous reports have identified tissue inversion post-transplantation. We describe inadvertent inversion of donor tissues during preservation and transportation from an eye bank. It is recommended to check the presence of corneo-scleral rim, trabecular meshwork, grooves, and residues of choroid if present inside or outside before transplantation to ensure accurate grafting and avoid transplantation failures.
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http://dx.doi.org/10.5301/ejo.5000711DOI Listing
February 2016

Amniotic membranes in ophthalmology: long term data on transplantation outcomes.

Cell Tissue Bank 2016 Mar 11;17(1):51-8. Epub 2015 Jul 11.

Veneto Eye Bank Foundation, Via Paccagnella 11, 30174, Venice, Italy.

The use of amniotic membrane (AM) is a widespread clinical practice for eye surgeries and the treatment of an increasing number of ocular surface pathologies. Here we describe the AM collection methods and donor selection criteria adopted by our tissue bank to distribute 5349 amniotic membrane patches over the last 12 years for the treatment of several ocular pathologies. Specific quality control measures are described and the long term results attained using the reported procedure are presented. A case of AM utilized to treat severe ocular ulceration is also described as an example of AM transplantation. Collective data for the total amniotic membrane patches deployed to treat various ocular diseases are discussed and success rates for AM transplantations are reported. An extensive follow-up is illustrated. The results suggest that the procedures and protocols used by the Treviso Tissue Bank Foundation and Veneto Eye Bank Foundation for collection, preservation, distribution and follow-up are of an optimal standard. Accordingly, the authors conclude that the safety and efficiency of the proposed procedure for the therapeutic use of AM to treat various ocular pathologies are reproducible, with additional evidence favoring the use of AM as an alternative to conventional medical treatment for certain ocular conditions.
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http://dx.doi.org/10.1007/s10561-015-9520-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4786589PMC
March 2016

A superfusion apparatus for ex vivo human eye irritation investigations.

Toxicol In Vitro 2015 Oct 20;29(7):1619-27. Epub 2015 Jun 20.

International Centre for Ocular Physiopathology, The Eye Bank Foundation of Veneto, Via Paccagnella 11, Padiglione Rama, Venice 30174, Italy.

A superfusion apparatus (SA) was developed to maintain isolated human corneas ex vivo under conditions which mimic the natural eye environment in vivo, including controlled temperature, tear flow and intraocular pressure. The SA was designed, developed and tested for use in ophthalmic pre-clinical research and to test new pharmaceutical formulations. Corneas undergo an equilibration process in the new physiological environment for one day. The test was then initiated by the application of the test substance, incubation, and temporal assessment of corneal damage using various parameters. The effects of mild and severe irritant concentrations of NaOH (2% and 8%, respectively) on corneal opacity, swelling and epithelial integrity were studied, and the inflammatory status assessed using F4/80 and MPO as macrophages and neutrophils markers, respectively. The SA was then used to test new artificial tear formulations supplemented with silver ions as an active constituent, showing different degrees of inflammatory responses as indicated by the migration of MPO and F4/80 positive cells towards the epithelium. The human cornea superfusion apparatus was proposed as a model for acute eye irritation research.
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http://dx.doi.org/10.1016/j.tiv.2015.06.022DOI Listing
October 2015

Surgical Excision of Orbital Progressive Granular Cell Tumour.

Case Rep Ophthalmol Med 2015 18;2015:420490. Epub 2015 May 18.

The Department of Maxillofacial Surgery, Ospedale dell'Angelo, Via Paccagnella 11, 30174 Venice, Italy.

Granular cell tumour (GCT) is mostly benign lesion first described by Abrikossoff and named after him. Most cases are reported in the head and neck area, where the tongue is the most common site. Here we review previous cases in the literature for GCT in the orbit and present a new case. A 49-year-old male presented with apparent exophthalmos. Examination of the patient revealed the presence of a mass in the bottom side of the orbit. A substantial progress was noted after two months from the initial examination using computed tomography (CT) scan. An orbital mass was extracted and histological analysis showed signs typical for GCT. Immunohistochemistry was positive for S-100; the biopsy showed no mitotic or necrotic areas. Proptosis was resolved after surgery and a six-year follow-up CT scan was performed. We conclude that rapid progress of the tumour does not necessarily suggest malignancy.
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http://dx.doi.org/10.1155/2015/420490DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4451783PMC
June 2015

Targeting herpetic keratitis by gene therapy.

J Ophthalmol 2012 26;2012:594869. Epub 2012 Dec 26.

The Veneto Eye Bank Foundation, Via Paccagnella 11, Padiglione Giovanni Rama, Zelarino, 30174 Venice, Italy.

Ocular gene therapy is rapidly becoming a reality. By November 2012, approximately 28 clinical trials were approved to assess novel gene therapy agents. Viral infections such as herpetic keratitis caused by herpes simplex virus 1 (HSV-1) can cause serious complications that may lead to blindness. Recurrence of the disease is likely and cornea transplantation, therefore, might not be the ideal therapeutic solution. This paper will focus on the current situation of ocular gene therapy research against herpetic keratitis, including the use of viral and nonviral vectors, routes of delivery of therapeutic genes, new techniques, and key research strategies. Whereas the correction of inherited diseases was the initial goal of the field of gene therapy, here we discuss transgene expression, gene replacement, silencing, or clipping. Gene therapy of herpetic keratitis previously reported in the literature is screened emphasizing candidate gene therapy targets. Commonly adopted strategies are discussed to assess the relative advantages of the protective therapy using antiviral drugs and the common gene therapy against long-term HSV-1 ocular infections signs, inflammation and neovascularization. Successful gene therapy can provide innovative physiological and pharmaceutical solutions against herpetic keratitis.
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http://dx.doi.org/10.1155/2012/594869DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3541562PMC
January 2013