In Alzheimer disease (AD) patients, amyloid β (Aβ) peptide-mediated degeneration of cholinergic system utilizing acetylcholine (ACh) for memory acquisition is observed. Since AD therapy using acetylcholinesterase (AChE) inhibitors are only palliative for memory deficits without reversing disease progress, cell-based therapeutic approaches are attracting attention. To recover cognitive function and to eliminate the causative Aβ peptides, we established F3.ChAT human neural stem cells (NSCs) encoding choline acetyltransferase (ChAT) gene (an ACh-synthesizing enzyme) as well as HMO6.NEP human microglial cells encoding neprilysin (NEP) gene (an Aβ-degrading enzyme) and HMO6.SRA cells encoding scavenger receptor (SRA) gene (an Aβ-uptaking enzyme). The cells (2 ⅹ 105 cells) were transplanted intracerebroventrocularly to mice showing memory loss induced by cholinotoxin AF64A, and brain Aβ accumulation, ACh concentration, and cognitive function were analyzed. Transplanted NSCs and microglial cells were found to express their functional genes in the mouse brain and restored the learning and memory function of AF64A-challenged mice by eliminating Aβ deposit and recovering ACh level, in which the effects were further enhanced by combinational treatments with F3.ChAT and HMO6.NEP or HMO6.SRA cells. The cell therapy also attenuated inflammatory astrocytic response by reducing Aβ accumulation. Taken together, it is expected that NSCs and microglial cells over-expressing ChAT, NEP or SRA genes could be candidates for replacement therapy of AD.
Prof. Yun-Bae Kim has completed his PhD at the age of 33 years from Seoul National University, Korea, and has studied as a visiting professor at University of British Columbia, Canada. He is a professor of College of Veterinary Medicine, Chungbuk National University, Korea. He has published more than 300 papers in reputed journals and has been serving as an Editor-in-Chief of Laboratory Animal Research.
Introductions.Despite earlier works ascribe the therapeutic effects of human mesenchymal stromal cells (MSCs) to their ability to engraft and differentiate into tissue resident cells, growing literature evidences have brought the attention to the array of moleculesproduced by MSCs. Cell secretome is a mixture of proteins, microvesicle/exosomes and genetic materials collected in a form of conditioned medium from cultured cells.Animal studies have shown that MSC-derived secretome promotes functional recovering in several districts, such as cardiac and neural, thus recapitulating the effects of MSC-based therapy. Particularly, growth factors, cytokines, and chemokines seem in an exclusive position as important modulators of MSC-mediated tissue repair. Methods.We used aTaqMan human angiogenesis array to analyze the expression of tissue repair-related genes in fetal dermal cells and their adult counterparts. Luminex multi-analyte profiling (xMAP) technology was employedto quantify growth factors and chemokines known for theirbeneficial action on tissue repair in cell secretome.Functional evaluation of cell secretomewas doneby testing its ability in inducingformation of capillary-like structuresof target cells (in vitro angiogenesis assay). Results.We found that upregulated genes (≥ 2-fold) in fetal vs adult samples were mostly linked to blood vessels formation and organization, while genes correlated with inflammation, particularly those regulating cell motility and recruitment, were significantly downregulated. Quantification of soluble factors revealed that fetal dermal cells secretome contained growth factors and chemokines such as VEGF-A, HGF, bFGF, IL-8, andGro-alpha detected at significantly higher values than those detected in adult dermal cells secretome.Moreover, fetal dermal cells secretomeinduced formation of capillary-like structures with higherefficiency than adult dermal cells.
After graduating in Natural Sciences in 1996 at the University of Palermo, he worked for several years in the field of Innate Immunity. In 2003 he obtained a Ph.D. in Animal Biology at the University of Palermo developing a thesis on humoral and cell-mediated mechanisms that regulate the inflammatory response in marine invertebrates, in collaboration with the Laboratory of Animal Physiology & Evolution of the Zoological Station A Dohrn of Naples and with the Pathology & Laboratory Medicine of the School of Medicine, University of Pennsylvania, Philadelphia (USA). In 2003 she was Visiting Scholar for a year at the Pathology & Laboratory Medicine of the School of Medicine, University of Pennsylvania, focusing on the role of complement in the inflammatory response in mouse models. From July 2004 to early 2006 she was a post-doctoral fellow at the Pharmacology Department of the School of Medicine, University of Pennsylvania, dealing with the expression of enzymes modulating the oxidative stress of the brain in transgenic mice suffering from "Alzheimer's disease-like" amyloidosis. "Since September 2006 he has been Research Associate at the Regenerative Medicine and Advanced Biotechnology Unit of the Fondazione Ri.MED of Palermo, where he is involved in several projects and since January 2013 he has been a Senior Researcher at the same Unit.
The injury of peripheral nerves is a common condition and damages both motor and sensorial functions. This injury especially is seen in motor vehicle accidents, falls or various trauma conditions. In contrast to central nervous system, the peripheral nervous has regeneration capacity after injury but this regeneration is quite poor. After the peripheral nerve injury, some cellular and molecular changes occur at the injury site such as demyelination or axonal degeneration. Myelin is very important for rapid axonal conduction and myelination process orchestrated by Schwann cells in peripheral nerves. Recently, studies have focused the enrichment of regeneration process by adding variety material into the lumen in this environment. Lithium is a mood stabilizer and is widely used as neuroprotective agent. It is an also enzymatic inhibitor of Glycogen synthase kinase 3β (GSK3β) and is mimics Wnt/β-catenin signal pathway. Wnt activation inhibits GSK3β and supress β catenin degradation. In addition, GSK3β inhibition speeds both debris clearance and remyelination. In our study, experimental peripheral nerve injury made on rat sciatic nerves and bridged with silicone conduit. Study protocol was approved by the Animal Ethics Committee of Eskisehir Osmangazi University (ESOGU-HADYEK approval No. 21.12.2016/568). This conduit filled with at different dosages lithium dilution. After three months, biopsy samples were prepared for confocal microscopic analysis. Our data suggested topical lithium stimulated the expression of Cyclin D1 and β catenin proteins. Confocal microscopic analysis verified the topical lithium administration showed positive effects in healing of injured peripheral nerve and remyelination. Acknowledgements: This work was supported by a grant from ESOGU-BAP (Project no: 2017-1483).
Ilknur Dag completed his PhD from Gazi University, Faculty of Science, TURKEY and doctoral studies from Eskişehir Osmangazi University, Medical Faculty, TURKEY. She studies in Eskişehir Osmangazi University, Central Research Laboratory, Application and Research Center, especially in electron microscopy area. He has published more than 20 papers in reputed journals.
The influence of biologically active additives, crosslinking agents, and enrichment with growth factors on the morphological properties of collagen-based scaffolds and their in-vitro bioactivity with mouse fibroblasts 3T3 have been investigated. 3D porous collagen scaffolds were modified with both antibacterial natural polysaccharides (chitosan and oxidized cellulose) and growth factors delivered in the form of blood platelet lysate. Addition of blood platelet lysate decrease the pore size of pure collagen scaffold but increased its porosity as demonstrate scanning electron microscopy pictures. Surprisingly, in-vitro tests of prepared scaffolds on mouse fibroblasts 3T3 showed that scaffolds enriched with platelet lysate exhibited significant synergistic effect with antibacterial polysaccharide-based additives on cells cultivation as determined by MTS assay and PicoGreen method. It is probably due to the cationic groups, either protonated amino groups or calcium cations from chitosan and oxidized cellulose, respectively, which are bioadhesive and readily binds to negatively charged surfaces and might therefore activated the blood platelets going along with acceleration of fibrin production. Moreover, angiogeneic potential of newly prepared scaffolds was proven by Chicken Chorioallantoic Membrane assay. Therefore, these newly developed antibacterial collagen sponges involving growth factors could be used as scaffold for growing cells in systems with low mechanical loading having potential application in soft tissue engineering.
Johana Babrnáková has completed her Master degree at Brno University of Technology, Czech Republic and 2 years ago started PhD study at Central European Institute of Technology, Czech Republic in the Advanced materials program. She has experiences in chemistry and materials science specifically biopolymers for regenerative medicine. During her Freemover internship stay at Sheffield University, UK, she has been trained in cell seeding and in-vitro characterization of biomaterials. Simuntaneously, she is engaged in the development and manufacturing of human and veterinary diagnostics.