Barbara Adinolfi Received The Master Degree In Pharmaceutical Chemistry And Technology In 2000 From The University Of Pisa, Italy. She Received The Phd In Pharmacotoxicological, Pharmacognostical Sciences And Pharmacological Biotechnologies In 2005 From The University Of Milan And The Specialization In Pharmacology In 2008 From The University Of Pisa. She Has Worked From 2000 Until 2013 At The Department Of Pharmacy, University Of Pisa. She Also Spent A Period (2009-2010) In The Department Of Biomedical Sciences, University Of Copenhagen. At Present She Is At The CNR Institute Of Applied Physics In Florence. Her Research Activity Is Focused On The Evaluation Of Anticancer Activity Of Natural And Synthetic Compounds, On The Evaluation Of The Role Of Specific Proteins As Putative Prognostic Markers In Cancer And On The Activity Of Nanoparticles As Intracellular Delivery Tools For Oligonucleotide Optical Switches By Confocal Microscopy.
Statement Of The Problem: The Development Of Effective Drug Delivery Systems Represents One Of The Main Goals In Nanomedicine. Data In Literature Demonstrate That, Depending On The Uptake Pathways, The Nanocarriers And Their Payloads Can Be Trafficked Into Different Organelles. In This Context, The Aim Of The Work Was To Evaluate: A) The Ability Of Polymethylmethacrylate Fluorescent Nanoparticles (PMMA-Nps) To Promote, In Human A549 Cancer Cells, The Internalization Of A Molecular Beacon (MB) Specific For Survivin Mrna As Theranostic Agent; B) The Involvement Of Endocytosis In The NP Uptake; C) The NP Fate At Different Times Of Cell Incubation To Verify Their Localization In Lysosomes; D) The MB Localization In Relation With The Endoplasmic Reticulum (ER) Where The Target Mrna Is Located. Methodology: The PMMA-NP Capability To Promote The MB Uptake By Endocytosis And The NP And MB Intracellular Localization Studies Were Realized By Confocal Microscopy Using Markers Of Endocytic Process And Organelles; The NP Fate Was Also Evaluated By Fluorescence Measurements. Findings: A) PMMA-Nps Promote The MB Uptake; B) Significant Increase Of The Number And The Mean Area Of Dextran Filling Endocytic Vesicles In The Presence Of Nps After Incubation Of 30'; C) Strong Co-Localization Of NP And Lysosome Tracker Fluorescence After 2-48 Hours Of Incubation (The Cell Culture Medium Fluorescence Decreases In The 24-72 Hour Window And Increases After 96 Hours); D) Co-Localization Of The MB Fluorescence With The ER-Marker Signal After 90' Of Incubation. Conclusion & Significance: These Data Highlight The Ability Of The PMMA-Nps To Promote The Survivin-MB Internalization And The Involvement Of Endocytotic Pathway In Their Uptake. These Results Demonstrate That The PMMA-Nps Are An Appropriate Delivery System Capable Of Being Eliminated By The Cells Involved In The Treatment; These Evidences Also Contribute To Consider The MB As An Effective Tool For The Intracellular Sensing.
Esther Carls Has Her Primary Research Interest In Cardiovascular Diseases. She Has Completed Her Bachelor Degree In Life Sciences In 2011 And Her Master Degree In ‘Clinical Molecular Sciences’ In 2012 Focusing On Macrophage Cell Therapy In Atherosclerosis. Following This She Worked At The University Of Edinburgh On Macrophage Cell Therapy In Renal Ischemia. Since 2014 She Has Been Working At The University Hospital Bonn’s Dept. Of Cardiac Surgery On Cell And Gene Therapy In Myocardial Infarction. In 2017 She Started Work On Her Phd Thesis Under The Supervision Of Prof. Dr. Med. Wilhelm Röll.
After myocardial infarction, one of the major complications, besides a reduced ejection fraction (EF), is the occurrence of ventricular tachycardia (VT). The loss of cardiomyocytes in the infarction area is structurally compensated by their replacement with fibroblasts during scar formation. The massive loss of Cx43 expression within the infarct zone hampers electrical conduction and favors ventricular tachycardia (VT) by re-entry mechanisms. Therefore we hypothesized that intramyocardial transplantation of Cx43 expressing embryonic cardiac fibroblasts (eCF) following myocardial infarction could reduce post-infarct VT incidence. eCF were harvested on E13.5 and cultured for 7 days and transduced with either a Cx43-IRES-GFP or an IRES-GFP lentivirus (LV) construct in vitro. Subsequently they were loaded with fluorescently labelled, PMAO coated magnetic nanoparticles (MNP) overnight. Mice (female CD1, 10-12 wks.) underwent cryogenic induction of myocardial infarction and 200.000 cells were subsequently injected into the infarction area, either with or without application of a magnetic field. Functional analyses were performed 14 days post op by echocardiography and electrophysiological examination. Injected eCF were retained in the infarction area as shown by GFP expression of fibroblasts in the infarction area as well as the presence of the MNP. In the Cx43 transduced eCF injected mice an increased expression of Cx43 in fibroblasts could be observed. Functional examination revealed significantly increased anterior wall thickening (AWT) and improved arrhythmia resistance in mice injected with Cx43 expressing eCF (+Magnet, 25.0% VT incidence, n=8) compared to mice injected with GFP control virus (+Magnet, 77.7% VT incidence, n=9). However, there were no significant differences in global left ventricular function (fractional shortening) between groups. In conclusion, the transplantation of Cx43 expressing eCF protects against VT after myocardial infarction, but does not improve heart function.