Regenerative treatment using a radioelectric asymmetric conveyor as a novel tool in antiaging medicine: an in vitro beta-galactosidase study

Clinical Interventions in Aging Dovepress open access to scientific and medical research O riginal R esearch Clinical Interventions in Aging downloaded from https://www.dovepress.com/ by 5.135.254.153 on 12-Jul-2018 For personal use only. Open Access Full Text Article Regenerative treatment using a radioelectric asymmetric conveyor as a novel tool in antiaging medicine: an in vitro beta-galactosidase study This article was published in the following Dove Press journal: Clinical Interventions in Aging 27 June 2012 Number of times this article has been viewed Salvatore Rinaldi 1,2 Margherita Maioli 1,3,4 Sara Santaniello 3,4 Alessandro Castagna 1,2 Gianfranco Pigliaru 3,4 Sara Gualini 3,4 Matteo Lotti Margotti 5 Arturo Carta 6 Vania Fontani 1,2 Carlo Ventura 1,4,7 Department of Regenerative Medicine, Rinaldi Fontani Institute, Florence; 2Department of Neuro Psycho Physio Pathology and Neuro Psycho Physical Optimization, Rinaldi Fontani Institute, Florence; 3 Department of Biomedical Sciences, University of Sassari, Sassari; 4 Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Bologna; 5Department of Information Technology and Statistical Analysis, Rinaldi Fontani Institute, Florence; 6Ophthalmology Section, University of Parma, Parma; 7 Cardiovascular Department, S Orsola Malpighi Hospital, University of Bologna, Bologna, Italy 1 Correspondence: Salvatore Rinaldi Rinaldi Fontani Institute, Viale Belfiore 43, 50144 Florence, Italy Tel +390 5529 0307 Fax +390 5529 0399 Email submit your manuscript | www.dovepress.com Dovepress http://dx.doi.org/10.2147/CIA.S33312 Powered by TCPDF (www.tcpdf.org) Background: Beta-galactosidase is the most widely used biomarker for highlighting the processes of cellular aging, including neurodegeneration. On this basis, we decided to test in vitro whether a set of rescuing/reparative events previously observed by us in subjects treated with radioelectric asymmetric conveyor (REAC) technology may also involve antagonism of a marker of aging-related degenerative processes, as assessed by a reduction in beta-galactosidase at the cellular level. Methods: Human adipose-derived stem cells were cultured at different passages, ranging from 5 to 20, with or without REAC exposure for 12 hours. The cells were then processed for biochemical beta-galactosidase staining and morphological microscopy analysis. Results: We observed a significant reduction in expression of senescence associated-betagalactosidase, and a persistence of fibroblast-like morphology typical of human adipose-derived stem cells, even at late passages. Conclusion: Our results indicate the ability of REAC technology to counteract in vitro senescence of human adipose-derived stem cells, and prompt the hypothesis that such technology may be exploited to antagonize in vivo senescence of tissue-resident or transplanted stem cells playing an important role in clinical treatment of age-related processes. Keywords: aging, adipose-derived stem cells, neurodegenerative diseases Introduction Aging of the human population is a problem with many social and economic implications. If aging is accompanied by disease, especially of the neurodegenerative type, the social and economic costs will increase even more dramatically. The aging processes are natural phenomena, but are accelerated and aggravated by environmental factors, often beyond the control or knowledge of the subject. It is also very difficult to implement real and effective strategies that can slow down the processes of aging and related diseases in a “biological” manner. On the other hand, biomarkers are presently available for close monitoring of the aging process. One of the most often cited in the literature for highlighting both the processes of cellular aging and neurodegeneration is beta-galactosidase.1–4 We have previously shown that radioelectric asymmetric conveyer (REAC) technology, using specific protocols, is effective in eliciting reparative phenomena and is able to drive gene expression profiles controlling stem cell differentiation and pluripotency in vitro. Based on these findings, the purpose of this study was to verify whether REAC technology, using a specific protocol known as the “in vitro regenerative treatment protocol” (IVRTP), may be able to influence the in vitro production of beta-galactosidase from human adipose-derived stem cells Clinical Interventions in Aging 2012:7 191–194 191 © 2012 Rinaldi et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited. Dovepress Rinaldi et al which have been subjected to an aging process throughout multiple passages in culture. Clinical Interventions in Aging downloaded from https://www.dovepress.com/ by 5.135.254.153 on 12-Jul-2018 For personal use only. Materials and methods REAC technology for therapeutic use REAC is an innovative technology5,6 involving biostimulation and/or bioenhancement techniques that induce weak radioelectric currents in tissues, thereby inducing cell reprogramming activity. The model used in this study (ASMED, Florence, Italy) is specific for regenerative treatment. REAC technology has demonstrated efficacy in ameliorating several stress-related disorders,7–13 depression,12,14,15 anxiety,12,15 social anxiety,16 agoraphobia,17 bipolar disorder,18 behavioral and psychiatric symptoms in Alzheimer’s disease,19 and impaired motor control.20–24 Recently, REAC technology using IVRTP has also demonstrated an ability to induce stem cell pluripotency and differentiation.25 In vitro regenerative REAC protocol REAC IVRTP consists of a sequence of radiofrequency bursts 250 msec in duration, with an off interval of 2.5 seconds. The REAC apparatus is placed into a CO2 incubator, set at a frequency of 2.4 GHz, and its conveyor electrodes are immersed into culture medium containing human adiposederived stem cells. The REAC-radiated power is about 2 mW, the electric field is 0.4 V/m, the magnetic field is 1 mA/m, the specific absorption rate 0.128 µW/g, and the density of radioelectric current flowing in the culture medium (J) during the REAC single radiofrequency burst is 30 µA/cm2. Isolation and culture of human adipose-derived stem cells According to the procedure approved by the local ethics committee, all tissue samples were obtained after informed consent. Human subcutaneous adipose tissue samples were obtained during lipoaspiration or liposuction procedures. After washing, the lipoaspirates were digested with 0.2% collagenase A type I solution (Sigma-Aldrich, St Louis, MO) under gentle agitation for 45 minutes at 37°C, and centrifuged at 2000 rpm for 10 minutes to separate the stromal vascular fraction from the adipocytes. If necessary, the mesenchymal stem cell fraction was treated with red blood cell lysis buffer for 5 minutes at 37°C, then centrifuged again. Th (...truncated)