OPHTHALMIC FORMULATIONS PISA
OPHTHALMIC FORMULATIONS: CHALLENGES AND ADVANCES
OPENING SPEECH
Patrizia Chetoni
Department of Pharmacy - University of Pisa
Introduction
Claudio Bucolo
Department of Biomedical and Biotechnological Sciences - Center for Research in Ocular Pharmacology (CERFO) - University of Catania
Prof. Claudio Bucolo is Full Professor of Pharmacology at the University of Catania. Prof. Claudio Bucolo held, in the past, the position of Visiting Scholar in Pharmacology at the Eye Research Institute of the Oakland University (Rochester, MI, USA).
He was the first Italian to receive the international "Dr. Roger Vogel Award for Pharmaceutical Research 2015” for the significant contribution in the field of research of new drugs for the treatment of ophthalmic diseases. In 2014 he received the "Silver Fellow Medal" of the Association for Research in Vision and Ophthalmology and in 2019 he received the "Gold Fellow Medal" with the following motivation: "In honor of outstanding service to the eye and vision community”. In 2019, the Neuroscience Center of Excellence of Louisiana State University awarded Prof Bucolo the prestigious "Chancellor's Award Guest" award with the following motivation: "For exceptional contributions to pharmacology applied to ophthalmology and outstanding academic leadership". Prof. Bucolo research mainly concern the field of ocular pharmacology and neuropharmacology. Prof. Claudio Bucolo is the author of over 200 scientific articles published in peer-reviewed journals and book chapters.
During his career he has filed 11 international patents in the field of pharmacological research. He is a reviewer of Ministry of Education, University & Research and Agence National de la Recherche projects and is Associate Editor of scientific journals in the field of pharmacology. From 2012 to 2013 he was President of the Pharmacology Section of the Organizing Committee of the ARVO (Association for Research in Vision and Ophthalmology, Rockville, MD, USA- Association with about 12000 members) annual conference and was re-elected in 2016 as a member of the ARVO Conference Organizing Committee; from May 2018 until May 2019 he again held the role of President of the Pharmacology Section of the Organizing Committee of the ARVO annual Conference; from 2004 to 2008 he was board of trustee of the Association for Ocular Pharmacology and Therapeutics and re-elected in March 2019 again as board of trustee. In addition, he organized in the past 20 years a series of European Frontiers Lectures delivered by outstanding European scientists and the annual IT-ARVO meeting on ocular pharmacology. He was Coordinator of the International PhD Program in Neuroscience from 2019 to 2023.
Prof. Bucolo is currently Deputy Rector for Education in PhD Programs and Masters; he also acts as liaison between the BIOMETEC Department and the School of Excellence of the University of Catania.
Since 2017, Prof. Bucolo is Director of the Research Center in Ocular Pharmacology of the University of Catania (CERFO).
Unmet needs in ophthalmology: pharmacological issues
The talk will be focused on unmet needs in ophthalmology from a pharmacological perspective. Ocular diseases such as diabetic retinopathy, age-related macular degeneration and glaucoma will be discussed to assess the pharmacological issues. Remarkable progress has been done so far in the ocular field in terms of drugs and innovative delivery systems. Drug discovery is one of the most interesting and challenging effort facing the pharmacologists. New challenges await us in the next future.
Massimo Dal Monte
Department of Biology - University of Pisa
Education and career
June 2018-present: Full Professor of Physiology, Department of Biology, University of Pisa, Italy.
2014-2018: Associate Professor of Physiology, Department of Biology, University of Pisa, Italy.
June 2009-November 2009: Visiting Fellow, Department of Molecular Genetics, German Institute of Human Nutrition Potsdam-Rehbruecke, Nuthetal, Germany.
2006-2014: Researcher of Physiology, Department of Biology, University of Pisa, Italy.
2002-2005: Postdoc fellow, Department of Physiology and Biochemistry, University of Pisa, Italy.
2001-2002: Technician (D category), Department of Physiology and Biochemistry, University of Pisa, Italy.
1998: Specialization in Biochemistry and Clinical Chemistry, University of Pisa, Italy (magna cum laude)
1996-2000: Postdoc fellow, Department of Physiology and Biochemistry, University of Pisa, Italy.
1996: PhD in Biochemistry, University of Florence, Italy.
1992-1996: PhD student, Department of Physiology and Biochemistry, University of Pisa, Italy.
1990-1992: Fellow, Institute of Mutagenesis and Differentiation, CNR, Pisa, Italy.
1990: Master degree in Biology, University of Pisa, Italy (magna cum laude).
Institutional and organizational activities
2023: Chair, 73rd National Meeting of the Italian Society of Physiology
November 2020-present: Director, Department of Biology, University of Pisa
September 2020-present: Board Member, Italian Society of Physiology.
2019-2020: vice-president of the Bachelor degree in Biotechnology, Department of Biology, University of Pisa, Italy.
2019-2020: vice-president of the Master degree in Molecular Biotechnology, Department of Biology, University of Pisa, Italy.
2018-2020: Member of the Scientific Committee of Area 05 – Biology, University of Pisa, Italy.
2015-2020: Coordinator of International Collaborations, Department of Biology, University of Pisa, Italy.
2015-2020: Erasmus+ Coordinator, Department of Biology, University of Pisa, Italy.
2015-2020: Chair of the General Physiology Unit, Department of Biology, University of Pisa, Italy.
2014-2019: Member of the Animal Welfare Board, University of Pisa, Italy.
2012-present: Member of the Board of the Ph.D. program in Molecular Medicine, University of Siena, Italy.
2012-2019: Responsible of the Animal Facility, Department of Biology, University of Pisa, Italy.
2012-2018: Member of the Scientific Committee of the Library of Natural and Environmental Sciences, Chemistry, Mathematics, Informatics and Physics, University of Pisa, Italy.
2012-2016: Member of the Scientific Committee of Area 05 – Biology, University of Pisa, Italy.
2009-2011: Member of the Board of the Ph.D. School in Biological and Molecular Sciences, University of Pisa, Italy.
2009-2011: Member of the Board of the Ph.D. School in Neurosciences and Endocrine-metabolic Sciences, University of Pisa, Italy.
2008-2010: Member of the Scientific Committee of Area 05 – Biology, University of Pisa, Italy.
2008-2012: Member of the Scientific Committee of the Library of Natural and Environmental Sciences, University of Pisa, Italy.
2007-2010: Member of the Board of Dean’s Advisor, Faculty of Sciences, University of Pisa, Italy.
2010: Supporting activities in the organization of the commemoration of the centennial anniversary of Moruzzi's birthday.
2009-2011: Member of the Board of the Ph.D. School in Basic and Developmental Neurosciences, University of Pisa, Italy.
2009-2012: Supporting activities in the organization of the Annual Meeting of Young Researchers in Physiology in Pisa (2009 and 2010) and in Sestri Levante (2011 and 2012).
2006-2008: Member of the Scientific Committee of Area 05 – Biology, University of Pisa, Italy.
2006-present: Member of the Departmental Council, Department of Biology, University of Pisa, Italy.
2004: Supporting activities in the organization of the 55th National Congress of the Physiological Society of Italy
Research activities
Role of neuropeptides in retinal physiology
Molecular mechanisms underlying retinopathy of prematurity
Functional role of the beta adrenergic system: retinopathy of prematurity, retinitis pigmentosa and melanoma
Functional role of beta-3 adrenoceptors as oxygen sensors
Molecular mechanisms underlying diabetic retinopathy
Functional role of the uPAR system in diabetic retinopathy
Molecular mechanisms underlying retinitis pigmentosa
Role of neuroinflammation in retinal diseases
Nutraceutics in the treatment of retinal diseases
Eye drops based on natural products to treat eye diseases
Exploring retinal function and dysfunction: animal models @DBio, UniPi
The Research Unit of General Physiology at the Department of Biology, University of Pisa has a long track record in studying the mammal retina, in both physiological and pathological conditions, using animal models of several human ocular pathologies (those of the retina, in particular) including retinopathy of prematurity, diabetic retinopathy, retinitis pigmentosa and glaucoma. Using approaches such as electroretinography and optical coherence tomography, we are able to follow the same animal longitudinally, enabling new information to be gathered that cannot be seen at a single timepoint. Matching electrophysiology, imaging, molecular and biochemical approaches we were, for instance, the first to explore the role of the beta adrenergic system in the retina, and to link this system to the development of retinopathy of prematurity, one of the main blinding diseases in newborns. The same approaches were also used in investigating the effect of nutraceuticals in reducing or preventing retinal dysfunction in diabetic retinopathy or in glaucoma. In this talk, we will resume some of the main results obtained in the last fifteen years in our labs, extending the view on the near future, represented by the ongoing studies on retina neurodegeneration in animal models of neurodegenerative diseases such as Alzheimer’s disease and amyotrophic lateral sclerosis.
Laura Lucarini
Department of Neuroscience, Psychology, Drug Research and Child Health (NEUROFARBA) - University of Florence
Academic achievements
PhD in Drug Area and Innovative Treatments 2016-2018
Second PhD thesis at the Department of NEUROFARBA (Neuroscience, Psychology, Drug Research and Child Health), Pharmacology and Toxicology Section, University of Florence, Florence, Italy. Title of the thesis “New therapeutic targets for the treatment of inflammation and fibrosis in a murine model of bleomycin-induced pulmonary fibrosis”.
2nd level Specialization Degree in Biochemistry and Clinical Chemistry 2005-2009
Second level specialization post-degree course in Biochemistry (2005-2009) at the University of Florence, Italy. Title of the thesis “Genotype and phenotype correlation in patients affected by Marfan syndrome and related pathologies”.
PhD in Clinical Pathophysiology 2002-2004
Ph.D. thesis defense in 2005. Title of the thesis “Molecular defects of collagen type VI in Bethlem myopathy and Ullrich congenital muscular dystrophy: genotype-phenotype correlation”, at the University of Florence, Italy.
Degree in Biological Science 1993-2000
Degree (MSc) in Biological Sciences in 2000, at the University of Florence, Faculty of Science, Italy. Title of the thesis “Role of the alterations in Fibrillin-1 in vascular pathology”.
Present academic position
03/2020-present Laboratory Technician for Research
Department of NEUROFARBA (Neuroscience, Psychology, Drug Research and Child Health), Section of Pharmacology and Toxicology, University of Florence, Florence, Italy.
Research experience and professional activities
01/2013–02/2020 Research Associate Position
Dept. of NEUROFARBA, University of Florence, Florence, Italy. Supervisor: Prof. Emanuela Masini (2013-2018) and Prof. Patrizio Blandina (2019).
2013-present Contributing Scientist
Project on the "Study of novel drugs for the treatment of glaucoma in rabbit models”, in collaboration between Department of NEUROFARBA, University of Florence, Italy and the NICOX RESEARCH INSTITUTE Srl, Bresso Milan, Italy.
2013-2017 Contributing Scientist
Projects granted by Fondazione Cassa di Risparmio di Firenze (2013, 2014, 2015, 2016), on the study of glaucoma, lung fibrosis, neuropathic pain, dysmetabolic syndrome, at the Dept of NEUROFARBA, University of Florence, Italy.
2015-2017 Contributing Scientist
Project granted by Regione Toscana "Tagatose effects in the prevention of cardiac ischemic injury in the small animal with dysmetabolic syndrome induced by a fructose-rich diet (TAGIDISFRU)”
2016-2020 Contributing Scientist
Project PRIN 2015 (2015ZT9HXY), PI Prof.Carlo Riccardi of the University of Perugia, financed by the Italian Ministry of Education, University and Research (MIUR).
2009-2010 Associate Researcher
Molecular Cardiology Laboratory at Saint Boniface Research Centre, University of Manitoba, Winnipeg, Canada. Supervisor: Prof Ian M C Dixon. Project on the study of molecular mechanisms of cardiac fibrosis, matrix deposition in hearth failure and matrix remodelling of hearth tissue.
2005-2009 Post-doc position
Department of Medical and Surgical Critical Care, University of Florence, Florence, Italy. Supervisor: Prof. Guglielmina Pepe. The research project was on the study of molecular defects of TGFBR1 and TGFBR2 in Marfan syndrome and related pathologies.
2002-2004 Contributing Scientist
Member of the international research team for the project Myo-Cluster, FP5-LIFE QUALITY, EU, for the study of Bethlem myopathy and related neuromuscular disorders.
2003 Visiting Research Fellow. Thomas Jefferson University, Philadelphia, USA. Supervisor: Professor Mon-Li Chu. Molecular biology research on extracellular matrix proteins and functional consequences of collagen VI genes mutations on RNA and proteins.
Ocular Hypertension and Retinal Neuroprotection advances in rabbit models
Laura Lucarini, Silvia Sgambellone, Emanuela Masini
Glaucoma is the second leading cause of blindness, afflicting more than 60 million people worldwide with a discouraging estimate of 111 million in 2040. It is a degenerative disorder of the optic nerve characterized by a malfunction of ciliary processes and trabecular meshwork (TM). This damage causes an imbalance between production and outflow of aqueous humor (AH), generating an increase of intraocular pressure (IOP). The ocular hypertension (OHT), if untreated, provokes a reduction of local perfusion and chronic ischemia leading to a depletion of RGCs by autophagy and/or apoptosis. Currently, all treatment strategies are directed at lowering IOP, reducing AH production or increasing the outflow. A significant number of patients do not respond to therapies and several side effects occur. In recent years, an approach consisting of multi-targeted compounds is emerging; this strategy consists in the design of hybrid compounds that incorporate moieties able to interact at different biological levels. Among the numerous hybrid molecules under preclinical study, we focused on histamine H3R antagonists-nitric oxide (NO) donors hybrid compounds. The histaminergic system plays an important role in the regulation of IOP; in fact, H3R antagonists are effective in reducing OHT, preventing retinal ganglion cells (RGCs) loss by improving vascular performance of the ophthalmic artery and in reducing oxidative stress. NO is an endogenous messenger that stimulates soluble guanylyl cyclase and evokes de-novo cGMP biosynthesis. The stimulation of NO signalling pathway, directly or via administration of NO donor compounds, lowers IOP through the relaxation of the TM. Moreover, impaired ocular blood flow has been shown to result in insufficient oxygen and nutrient supply to the RGC axons crossing the optic nerve head (ONH), causing their degeneration. Indeed, unstable ocular blood flow, produced by chronically elevated and/or oscillating IOP, can lead to repeated hypoperfusion, resulting in ischemic damage.
The aim of this research was to evaluate the efficacy of histamine H3R antagonists-NO donor hybrid compounds in two rabbit models of OHT, and in one rabbit model of ophthalmic artery ischemia/reperfusion (I/R), evaluating their capability in reducing IOP, ameliorating vascular ocular performance and preventing photoreceptor degeneration.
The acute OHT model was obtained by injection of 50 µl of hypertonic saline (5% NaCl) into the vitreous, while the chronic one injecting 100 µl of carbomer into the anterior chamber of rabbits’ eye. The IOP was measured with a pneumotonometer at baseline, 60,120 and 240 min after acute OHT induction, and every day before drug dosing in the chronic model for 12 days. All animals received a topical dose of hybrid compound (30 μl, 1%) or vehicle or reference drugs, ciproxifan 1% or molsidomine1%. The I/R model was obtained by repeated injections of ET-1 twice a week for six weeks into the subtenon capsule of the eye. The animals were treated twice daily with vehicle or hybrid compound ST-1989 from the third week until the end of the protocol. IOP was measured once a week after 36 hours of washout. Ophtalmic artery resistence index and retinal functions were assessed using a Color Doppler ultrasound and Retimax Advance, respectively, at baseline, after 2 weekly injections of ET-1 and after 6 weeks. After 6 weeks, the animals were euthanized to collect whole eyes, AH, ciliary body and retina tissues. The hybrid compound ST-1989 was able to reduce the IOP in both acute and chronic OHT models demonstrating a long-lasting effect. It reduced inflammatory response (IL-6 and TNF-α), oxidative stress (GSH, MnSOD and 8OH2dG), and apoptotic process in chronic OHT and attenuated hypertension, improved ocular perfusion and prevented photoreceptor degeneration in the retinal I/R model. In conclusion, the hybrid compounds could be a potential therapeutic drug for glaucoma management and neuroprotection.
Fabio Neggiani
Farmigea SpA - Pisa
He has more than 27-year experience in the pharmaceutical industry and more than 20-years of roles of high responsibility in the R&D, CMC and Project management areas.
Miscellaneous know-how from the preformulation to the technology transfer for the industrial production and ranging from the New Chemical Entities to generics. Extensive and significant experience in a pharmaceutical developmental role. Strong technical experience in terms of early and later stage development of drug product or design of medical device. Strong analytical experience for the characterization of drug substance and quality control of drug substances and drug products.
During his working activity he has developed many products up to the launch on the market of different pharmaceutical forms as tablets, capsules, oral solutions, gels, creams, ointments, parenteral, eye drops and ointments.
Ophthalmic formulation for Dry Eye
Dry eye disease (DED) affects hundreds of millions of people throughout the world and is one of the most frequent causes of patient visits to eye care practitioners. Short review of the definition, the impact, classification & risk factors of dry eye disease. Highlight the evolution of therapeutic approaches and challenges for the treatment of dry eye disease. The history of Farmigea R&D in the dry eye field and the new DED project under development.
Sara Nicoli
Department of Food and Drug - University of Parma
Sara Nicoli received in 2001 her Ph.D. in Biopharmaceutics and Pharmacokinetics, from the University of Parma (IT) and the University of Paris XI (FR). Since 2014 she is Associate Professor at the Department of Food and Drug of the University of Parma. Her research is focused on innovative approaches to enhance dermal, transdermal and ocular drug delivery. She is author or co-author of more than 90 papers published on international peer-reviewed journals.
Overcoming ocular barriers using polymeric micelles
Polymeric micelles represent an interesting formulative approach for ocular drug delivery, as they act as solubility enhancers of poorly soluble drugs and can promote drug transport across conjunctiva, cornea and sclera. Despite their potentiality, several aspects deserve further clarification, mainly with respect to their stability, mechanism of action and interaction with ocular tissues. Indeed, drug delivery from this carrier depends upon several phenomena: drug release from intact micelles, micelle’s dissociation, interaction with the biological environment. In this presentation the attention will be mainly focused on the interaction of micelles with ocular tissues, to clarify the mechanisms of drug transport. Micelles studied are composed of a water-soluble derivative of vitamin E (TPGS: d-alpha tocopheryl polyethylene glycol succinate), alone or associated with other polymers, and loaded with different drugs used for the treatment of anterior and posterior segment eye diseases. Micelles interaction with ocular tissues has been investigated through ex-vivo studies, paying attention not only to the drug, its accumulation in the tissues and permeation through the membranes, but also to the fate of the polymer. The use of two-photon microscopy combined with the simultaneous quantification of the drug and the polymer, represents a useful tool for shedding light on the mechanism of micelle penetration and on the factors influencing the drug transport.
Rosario Pignatello
Department of Drug and Health Sciences - University of Catania
Prof. Rosario Pignatello is Full Professor of Pharmaceutical Technology and Legislation at the University of Catania, Italy. He is the actual Director of the Department of Drug and Health Sciences.
He has nearly 30 years of experience in the research and development of innovative formulations for the controlled release and targeting of bioactive molecules, through chemical approaches as well as nanotechnological carriers, aimed to treat different disorders.
Prof. Pignatello has coauthored about 180 papers and edited a series of textbooks on biomaterials and their application in biomedicine.
The main areas of research are polymeric and lipid-based micro- and nanoparticles as modified drug delivery systems; vesicular nanocarriers (liposomes, micelles); lipophilic prodrugs and conjugates; synthesis and evaluation of new polymeric biomaterials for drug delivery and tissue regeneration.
In particular, Prof. Pignatello works actively in the field of ocular drug delivery, leading the Research Centre for Nanomedicine and Pharmaceutical Nanotechnology (NANOMED) at the University of Catania.
Setting up topical lipid nanoformulations for diabetic retinopathy management: optimization and pharmacokinetic studies
Rosario Pignatello, Elide Zingale, Angelo Spadaro, Pradip K. Sasmal, Dhanoj Reddy Doni, Rajasekhar Jaladi, Basant Amarji, Ganesh Ramachandran, Shanavas Alikunju
Axitinib is a multi-receptor tyrosine kinase inhibitor molecule. It acts by blocking the receptors for vascular endothelial growth factor (VEGFR) and platelet-derived growth factor (PDGFR), which are responsible for the development of neovascularisation. To date, intravitreal injections of anti-VEGF have become the first-line medical treatment for diabetic macular oedema, a degeneration of diabetic retinopathy. In this category, anti-VEGF are able to induce regression of iris and retinal neo-vascularisation and reduce the risk of intraoperative and post-operative haemorrhage after vitreous surgery.
Despite the efficacy of intravitreal injections, the complications of this type of administration are manifold. Our study aims to design a nanoformulation capable of delivering Axitinib to the retinal tissue after topical administration. With the aim of increasing the aqueous solubility of this poorly soluble compound, two lipid-based formulations, namely NLC and SLN, were designed with the use of a design of experiment approach (DoE). The formulations were optimised and characterised, in particular focusing on loading with the highest possible dose of the drug, and further tested in vivo. Pharmacokinetic studies showed that both systems ensured a greater retinal targeting than a drug suspension. The NLC were more interesting in terms of the higher loaded drug concentration and amount of Axitinib reaching the retina.
Chiara Posarelli
Department of Surgical, Medical and Molecular Pathology and Critical Care Medicine - University of Pisa
She graduated in Medicine and Surgery in 2007 and specialized in Ophthalmology in 2012
Since January 2014 to December 2015
Grant, Glaucoma and uveitis
Azienda Ospedaliero-Universitaria Pisana
Since October 2014 to october 2016
Postgraduation Research Doctorate, Clinic Physiopathology
Azienda Ospedaliero-Universitaria Pisana
Since January 2016 to 31 August 2022
Researcher, Azienda Ospedaliero-Universitaria Pisana
Glaucoma and Uveitis
Since 1 September 2022 to date
Associate Professor at University of Pisa
Glaucoma and Uveitis
Personal skills:
Glaucoma diagnosis and treatment also laser treatment (Iridotomy, Iridoplasty, SLT)
Eye surgeon (Cataract, Glaucoma and Refractive Surgery)
Echographist Diploma SOU (2012)
Fellow of the European Board of Ophthalmology Glaucoma Subspecialty (2015)
Fellow of the European Board of Ophthalmology (2018)
Diploma of completion of the SOE Leadership Development Programme (Class 2021 -2023)
She is author of 92 papers (78 indexed peer reviewed international journals, 10 book chapters, 4 papers on national journals) and author of several personal communications at national and international meetings.
Surgical devices in glaucoma surgery: clinical evidences and future perspectives
Glaucoma is a leading cause of irreversible blindness and the second most common cause of blindness worldwide. According to the progressively increase and aging of the population, the number of glaucomatous patients is expected to grow up and it was estimated that in 2040 about 111.8 million people will be affected. To date, the lowering of the intraocular pressure (IOP) seems to be the only proven treatment to manage the disease. Different methods could be performed to lower the IOP such as medical, laser and surgical therapies. The chronic application of topical medications has been proved to be related with the development of dry eye disease, conjunctival inflammation and consequently poor adherence and compliance. In recent years, focusing on the surgical approach great progresses have been made, thanks to the introduction of new devices to reduce the complications of topical treatments. Sustained release implants and Micro-Invasive Glaucoma Surgery (MIGS) shared some characteristics: minimally manipulation of the target tissue, efficacy in lowering IOP, safety from serious complications, short learning curve for the surgeon and short recovery time for the patient.
Simona Sapino
Dipartimento di Scienza e tecnologia del farmaco - University of Turin
Simona Sapino received her Master Degree in Pharmaceutical Chemistry and Technology in 2000 her professional license as a pharmacist in 2001, and her Ph.D. in Pharmaceutical Science in 2006 from the University of Turin. Currently, as a Research Technician at the Department of Drug Science and Technology (University of Turin) she is involved in research activities within the scientific-disciplinary field CHIM/09 with the study of micro-nanosystems for delivery of drugs and functional substances. The study focuses on various vehicles such as emulsions, micro-emulsions, micellar solutions, liposomes, nanoparticles, cyclodextrins, mesoporus silica, etc. She also conducts studies on the determination of physicochemical parameters and in vitro tests with model systems to evaluate the release and efficacy of drug delivery systems. Recently, she has developed an in vitro model to perform test of ocular clearance on novel ophthalmic formulations. Her research activity is documented by more than 50 peer-reviewed scientific articles (H-index 23), 1 book chapter, 1 patent (WO 2010/058242) and several communications at scientific conferences and workshops. She is also involved in teaching activities by assisting students in the development of experimental thesis; moreover, she is part of the examination committees, as a subject matter expert, in the disciplinary field CHIM/09. She is teacher in the E.M.O.T.I.O.N Master (European Master on Translational Cosmetic and Dermatological Sciences).
She is affiliated with the Association of Italian Teachers and Researchers of Pharmaceutical Technology and Legislation (ADRITELF) and the NIS Interdepartmental Center.
In vitro ocular flow cell model enabling preliminary studies on new ophthalmic formulations
Simona Sapino, Giulia Chindamo, Marina Gallarate
Repeated intravitreal injections employed for the treatment of retinal diseases can result in complications and severe adverse reactions. The development of innovative drug delivery systems could obviate these drawbacks, but requires extensive research, including the use of in vitro models in preclinical trials. Starting from the literature, a Plexiglas in vitro eye model has been designed and developed in our laboratory, consisting in a 3-compartment ocular flow cell that emulates the anterior and posterior segments of the human eye. The model is filled with a simulated vitreous based on hyaluronic acid and agar; to mimic real conditions, a continuous aqueous flow is ensured by a peristaltic pump.
A recent improvement consisted of placing a collagen-based scaffold enriched with human retinal cells (ARPE-19) between the central and posterior compartments to simulate the human retina and thus to predict the possible drug-retina interactions. Avastin® (commercial bevacizumab) was injected into the central compartment as a test formulation. The encouraging results indicate the potential of the proposed in vitro model as a promising platform for evaluating the efficacy of novel ophthalmic formulations intended for the posterior segment eye treatment. Additionally, it could aid in reducing the use of animal experiments, particularly in the initial stages of development of a novel formulation.
Maria Tilde Torracca
Department of Drug - Livorno Hospital, Azienda USL Toscana nordovest
Maria Tilde Torracca obtained her degree in Pharmaceutical Chemistry and Technology in 1985 and went on to earn a PhD in "Design, development, and bio-evaluation of drugs" in 1993. In 1997, she further specialized in Pharmacology.
Since January 1999, Maria Tilde Torracca has been working as a Pharmacist in different Hospital Pharmacies. In April 2021, she assumed the position of head of the Pharmaco and Device Vigilance Unit at Azienda USL Toscana Nordovest.
Maria Tilde Torracca has been strongly involved in the galenic laboratory activities of Hospital Pharmacies since 1999. Her contributions led her to assume the role of coordinating the activities of all the laboratories of Azienda USL Toscana Nordovest in 2016.
In addition, Maria Tilde Torracca currently teaches "Pain Therapy: Legislation and Tecnological Aspects" as part of the Specialization School of Clinical Pharmacy at the University of Pisa.
Throughout her career, Maria Tilde Torracca has published numerous papers and has been involved as an author and co-author in various posters and oral communications.
Ophthalmic Preparations in Hospital Pharmacy: therapeutic opportunities and strategies
Maria Tilde Torracca, Marco Giannini, Elena Nicolai, Elisa Bertinetto, Laura Mariotti
Pharmacists play a crucial role in preparing sterile ophthalmic products when they are not readily available from licensed manufacturers. These products can be administered through various routes, including topical application, subconjunctival injection, and intraocular injection (such as intravitreal and intracameral routes). They can be formulated as solutions, suspensions, or ointments. In addition to addressing product availability, the preparation of ophthalmic products in-house also contributes to the economic sustainability of treating age-related macular degeneration.
To ensure that our hospital pharmacies meet national and international standards in ophthalmic preparations, we have conducted a quality assessment project. This project aims to evaluate and enhance the quality of our ophthalmic preparations, aligning them with established guidelines and regulations.
At present, our laboratory prepares several ophthalmic products, commonly used in the management of various eye conditions, including bevacizumab and mitomycin syringes for intravitreal injections. Additionally, we produce fortified eye drops with high concentrations of antibiotics to effectively treat keratitis. Moreover, we formulate cyclosporine or tacrolimus eye drops for patients suffering from conditions such as Sindrome sicca or Vernal Conjunctivitis.
In this regard, we will share the data regarding our activities and discuss the organization of our ophthalmic preparation processes. This will highlight the efforts we have made to ensure high-quality and standardized ophthalmic preparations, supporting optimal patient care in our hospital setting.
Mario Vergara
Ophthalmologist veterinary doctor
Nato a Lecce il 01/03/1966, laureato a Perugia il 06/11/1992.
Dal 1996 si occupa di oftalmologia, iscrivendosi alla SOVI (Società di Oftalmologia Veterinaria Italiana) e frequentandone i corsi e gli incontri nei vari anni.
Dal 2008 è socio di ANVO (Associazione Nazionale Veterinari per l’Oftalmologia), dal 2009 è Consigliere nel Consiglio Direttivo e dal 2021 ne è Presidente.
Ha svolto internship presso il Servizio di Oftalmologia dell’Unità dei Piccoli Animali dell’Ospedale Veterinario della UAB (Università Autonoma di Barcellona ES), diretto dalla Dr.ssa Teresa Peña e dalla Dr.ssa Marta Leiva e presso il Tieraerztliche Spezialisten del Dr. Jens Linek, Rodigallee 85 D- 22043 Hamburg Germany.
Dal 2019 al 2022 ha frequentato e completato l’Executive Master in Veterinary Ophthalmology diretto dal Dr. Rick Sanchez.
Nel corso degli anni ha presentato relazioni per varie Società Veterinarie (SOVI, ANVO, AVULP, ATOVELP, CogitoErgoVet, Circolo Veterinario Pugliese), ditte farmaceutiche e mangimistiche (Trebifarma, NBF, Monge), Ordini Professionali (PG, VT, AQ, LE), relatore ed Istruttore a vari Corsi di base ed avanzati ANVO e relatore presso il Dipartimento de Medicina Veterinaria di Perugia e di Camerino nell’ambito del Corso di Fisiologia Veterinaria nell’Anno Accademico 2021-2022 e 2022-2023.
Professore a contratto presso l’Università di Medicina Veterinaria di Bari nell’ambito della Scuola di Specializzazione in Malattie Infettive, Profilassi e Polizia Veterinaria nell’anno 2022.
Dal 2007 presta opera di consulenza esclusivamente su problematiche oftalmologiche, mediche e chirurgiche, presso varie strutture veterinarie italiane.
Ocular surface diseases: diagnosis and treatment
The ocular surface, as the first protective barrier, includes tears. The lacrimal system consists of a secretory and an excretory portion. The secretory portion comprises various glands with different histological morphology and secreted products, including two lacrimal glands, accessory glands, and the principal gland with a mixed mucous-serous secretion. Additionally, there are Meibomian glands that secrete lipids and goblet cells that secrete mucus.
The normal interaction of these glands enables the proper functioning of the ocular surface, serving optical, mechanical, defensive, and trophic functions. However, any dysfunction or alteration of these glands, leading to an imbalance in the tear film, can result in a condition known as keratoconjunctivitis sicca (KCS). This condition is characterized by corneal epithelium exposure and subsequent irritation.
There are two types of KCS:
-
Quantitative KCS: This occurs due to a reduced production of the aqueous component and can be caused by hereditary systemic pathologies or neurological pathologies (neurogenic KCS).
-
Qualitative KCS: This is attributed to a decreased lipid component, leading to increased evaporation of the aqueous part of the tear film. It can also result from a lower mucous component, which reduces adherence and permanence of the aqueous part on the corneal epithelial surface.
The therapeutic approach, following a thorough diagnostic examination with specialized instruments, may involve the use of different types of tear drops with varying compositions and viscosities based on the type of KCS.
Stefania Vernazza
Department of Experimental Medicine (DIMES) – University of Genoa
Work experience
01/12/2021-Current
Full-time fixed-term Researcher of General Pathology, Department of Experimental Medicine (DIMES), University of Genoa
The main lines of research are the following:
- To study the mechanisms underlying glaucoma pathogenesis and to investigate the mechanisms of mineral fiber-induced toxicity by taking advantage of advanced Human-based 3D culture systems
- To investigate the mechanisms of drug resistance in cancer cells
01/01/2021 – 30/11/2021
Fixed-term research contract with IRCCS G.B. BIETTI Foundation (IRCCS Fondazione G.B. Bietti per lo studio e la ricerca in oftalmologia), Rome
Participant in the research team involved in the project “Analisi di biomarkers associati alla patogenesi e alla progressione del glaucoma"
11/11/2019 – 30/09/2021
Fixed-term contract with OMIKRON ITALIA SRL
Research and consultancy activities for the study "3D-trabecular meshwork and neuron models to evaluate the biological potential of OMK1 eye drops in glaucoma therapy"
01/01/2018 – 31/12/2020
Co.Co.Co. (IRCCS Fondazione G.B. Bietti per lo studio e la ricerca in oftalmologia), Rome
Scientific research collaboration in the project "Study of the molecular basis responsible for the pathogenesis and progression of glaucomatous neurodegeneration", glaucoma operating unit.
01/01/2019-30/09/2020
Fixed-term contract with Eyepharma Spa. Research and consultancy activities for the study "3D model of trabecular meshwork for evaluation of DRAIN Drops biological potential in glaucoma treatment"
01/06/2017 – 30/11/2017
Research fellowship, Department of Experimental Medicine (DIMES), University of Genoa,"Alternative/substitute methods for animal testing".
Scientific organizations/coordination of academic activities
22-23/06/2022 Part of the scientific and organizing committee of the theoretical and practical course “Approcci innovativi in Ambito cosmetico, in ottemperanza con la regolamentazione europea”. CME credits provided.
29-30/09/2016 Part of the scientific and organizing committee of the eighth edition of the theoretical and practical course “colture cellulari: I metodi alternativi”. CME credits provided.
22-23/10/2015 Part of the scientific committee of the 2nd edition of the theoretical and practical course “DARE UN SENSO AI METODI ALTERNATIVI ALLA SPERIMENTAZIONE ANIMALE”. CME credits provided.
An advanced in vitro 3D platform as useful tool to study molecular change underlying glaucoma pathogenesis
Primary Open Angle Glaucoma (POAG) is an etiology complex and multistep disease, often characterized by trabecular meshwork (TM) dysfunction, increased intraocular pressure (IOP), and progressive vision loss.
Although several harmful IOP-independent mechanisms contribute to glaucomatous degeneration, IOP management is currently the only proven modifiable risk factor for treating glaucoma. In this regard, there is an increasing interest in designing novel multi- or single-target compounds to improve glaucoma drug-based treatment.
The use of advanced human-based in vitro models modeling the early stage of glaucomatous damage offer the possibility to understand the cellular and molecular mechanisms that contribute to the disease onset, thereby reducing the use of experimental animals. To provide an approach to this task, we performed a 3D in vitro model of human TM cells (HTMC) cultured in a milli-scale bioreactor system (IV Tech srl) to assess the cell responses to oxidative stress, which is considered the significant risk factor involved in TM pathological changes. To mimic the prolonged oxidative stress condition, the 3D HTMCs were treated with 500µM H2O2 2 hours a day up to 72hrs and the oxidative stress-induced effects were analyzed in terms of cell viability reduction, intracellular ROS production, apoptosis, cell senescence, and inflammatory markers. The bioreactor system is designed to provide a constant medium supply by stabilizing its influx to the culture chamber. Such an experimental platform provided more physiological 3D culture conditions than the standard static ones. The system-induced dynamism drove the HTMC adaptation to the applied stressor toward the expression of pro-inflammatory markers rather than cell death. Our in vitro model proved to be useful for testing the efficacy of active substances of two commercial eyedrops in restoring HTMC cells from oxidative stress-induced damage, suggesting them as TM-targeted therapy in addition to that already known on retinal ganglion cells.
Elide Zingale
Department of Drug and Health Sciences - University of Catania
Elide Zingale is a PhD student in Neuroscience at University of Catania, Italy, under the supervision of Professor Rosario Pignatello.
She received her Master's degree in Pharmacy in 2020. During her thesis, she frequented the laboratory of Pharmaceutical Technology (University of Catania) and the Italian Research Council (CNR) in Catania as a part of a thesis project on the “Optimization of curcumin nanocrystals by Box-Behnken Design for nose-to-brain delivery”.
In September 2020, he won a research fellowship for one year in a collaborative project with a pharmaceutical company (Dr.Reddy's Ltd.) on the development of lipid nanoparticles for the ocular delivery of molecules of industrial interest. Now Elide Zingale is focused on the field of ocular delivery with a doctoral project regard the "Development of different nanocarriers loaded with Sirt-1 agonists for the treatment of degenerative eye diseases".
Optimization by response surface methodology of self-nanoemulsifying drug delivery systems (SNEDDS) for ocular drug delivery
Elide Zingale, Angela Bonaccorso, Rosamaria Lombardo, Agata Grazia D’Amico, Velia D’Agata, Rosario Pignatello
Poor ocular bioavailability of many active molecules, combined with stability issues of formulated systems, prompt searching for innovative technologies. In this respect, self-nanoemulsifying drug delivery systems (SNEDDS), due to the presence of biocompatible and biodegradable ingredients and ease of large-scale production, combined with the opportunity for high drug loading, are of great interest. These systems allow to improve the solubility, stability and absorption of actives, resulting in greater bioavailability of highly lipophilic molecules. SNEDDS, consist essentially of three components in an anhydrous mixture: oil, surfactant, and co-surfactant. The choice of starting materials and the ratio in which they are combined is the crucial point for obtaining an optimal system suitable for ocular delivery. These systems can represent an advancement of actual micro- and nanoemulsion technologies, because they are emulsified directly in-situ, avoiding the problem of drug loss during storage.
The aim of the current study was to develop SNEDDS intended for ocular delivery of two different Sirt-1 agonists, Resveratrol (RSV) and Melatonin (MEL), for a potential support in diabetic retinopathy.
The preformulation phase was performed by ternary plot construction thanks to a DoE software using I-optimal design, to identify the emulsion zone. The optimization phase was performed using Response surface methodology (RSM). At this point, four batches of SNEDDS, with four different surfactants (Tween® 80, Tween® 20, Solutol® HS15, Cremophor EL®) were optimized. The stability study identified the system with Tween® 80 as the most stable and it was chosen for the following studies.
SNEDDS were prepared by a low energy method. Optimized platforms were prepared with Capryol PGMC, Tween® 80 and Transcutol® P and loaded with RSV or MEL. RSV-SNEDDS and MEL-SNEDDS were subjected to physico-chemical characterization, involving the simulation of their behaviour in the ocular environment, after reconstitution with simulated tear fluid (1/10; v/v) and under slight agitation to simulate blinking phenomenon.
Specifically, SNEDDS were evaluated for mean size (< 50 nm), homogeneity (PDI < 0.2), emulsion time (around 40 seconds), clarity by turbidimetry (about 100% transmittance), drug content (up to 90%), mucoadhesion studies, pH and osmolarity (both found close to physiological values), stability to dilution, cloud point and in vitro cytotoxicity studies on rabbit corneal epithelial cells (SIRC).
Optimized SNEDDS can be used as a promising nanocarrier for the ocular delivery of Sirt-1 agonist agents.