Gordana Nikcevic, Ph.D.is Senior Research Associate, Laboratory for Molecular Biomedicine at Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade, Belgrade, Serbia. She is also Scientific Coordinator/Assistant Coordinator of the EU FP7 Project (FP7-REGPOT: Strengthening the Research Potential of IMGGE through Reinforcement of Biomedical Science of Rare Diseases in Serbia – en route for innovation; SERBORDISinn). Her research interest are studying the molecular basis of various rare diseases, the regulation of eukaryotic gene expression and genetic variants relevant for the optimization of drug therapy in the treatment of inflammatory diseases. Studying the approaches to increase therapeutic capacity of animal mesenchymal stem cells.
Mesenchymal stem cells (MSCs) show enormous potential for cell-based therapy in the treatment of various diseases. In the present report, the genetic manipulation of canine MSCs isolated from adipose tissue has been investigated, with the aim of enhancing their therapeutic potential. Since MSCs are regarded as hard-to-transfect cells, we initially optimized the transient transfection efficiency of these cells using the LacZ reporter vector and in situ -galactosidase staining. Next, by applying optimized conditions, we transfected MSCs with pCIneoIL-10 plasmid that contains the cDNA of human IL-10, the potent immunosuppressive cytokine which is not produced by MSCs at a significant level. Using an ELISA assay, in which the level of human IL-10 from the supernatant samples of transfected canine MSCs was quantified, the successful expression of transgene was confirmed. The obtained results provide a working platform for further studies related to the reinforcement of MSCs-mediated therapeutic impact and the targeted delivery of various biological agents to disease sites.
Hung-Chih Hsu completed his MD degree from Taipei Medical University, and Ph.D. from Chang Gung University, Taiwan. He finished his residency at the Department of Physical Medicine and Rehabilitation (PM&R) at Chang Gung Memorial Hospital, and fellowship in Washington Medical Center in Seattle, USA. In addition to PM&R, he is also a specialty doctor of Geriatric Medicine, and Medical Diagnostic Ultrasound. He served as the director of PM&R, and Center of Developmental Pediatrics at ChiaYi Chang Gung Memorial Hospital. He is now the supervisory board of Taiwan Academy of Physical Medicine and Rehabilitation, Taiwan Myopain Society, Taiwan Sports Medicine Association. He is specialized in myofascial pain treatment, electrodiagnostic medicine, and regeneration medicine. He has published more than 50 papers in reputed journals and three books in his specialty.
Despite a plethora of literature has documented that osteoarthritis (OA) is veritably associated with oxidative stress-mediated chondrocyte death and matrix degradation, yet the possible involvement of synoviocyte abnormality as causative factor of OA has not been thoroughly investigated. For this reason, we conduct the current studies to insight into how synoviocytes could respond to an episode of folate-deprived (FD) condition. First, when HIG-82 synoviocytes were cultivated under FD condition, a time-dependent growth impediment was observed and the demise of these cells was demonstrated to be apoptotic in nature mediated through FD-evoked overproduction of reactive oxygen species (ROS) and drastically released of cytosolic calcium (Ca2+) concentrations. Next, we uncovered that FD-evoked ROS overproduction could only be strongly suppressed by either mitochondrial complex II inhibitors (TTFA and carboxin) or NADPH oxidase (NOX) inhibitors (AEBSF and apocynin), but not by mitochondrial complex I inhibitor (rotenone) and mitochondrial complex III inhibitor (antimycin A). Interestingly, this selective inhibition of FD-evoked ROS by mitochondrial complex II and NOX inhibitors was found to correlate excellently with the suppression of cytosolic Ca2+ release and reduced the magnitude of the apoptotic TUNEL-positive cells. Taken together, we present the first evidence here that FD-triggered ROS overproduction in synoviocytes is originated from mitochondrial complex II and NOX. Both elevated ROS in tandem with cytosolic Ca2+ overload serve as final arbitrators for apoptotic lethality of synoviocytes cultivated under FD condition. Thus, folate supplementation may be beneficial to patients with OA.