Regenerative medicine and stem cells

The stem cells offer hope for unprecedented care and perhaps the healing of tissues severely damaged that they cannot be saved even by the most advanced surgical or pharmacological treatments. This perspective has opened the way for a new paradigm in the management of complex diseases, the so-called “Regenerative Medicine”, which has the potential to cure old diseases and help a healthy and active aging, with exceptional socio-economic impact.

The field of Regenerative Medicine and Stem Cell has received a major boost from the recent observation that using gene transfer with viral vectors with only three or four genes, it may be possible to reprogram human somatic adult cells non-stem cells (i.e. fibroblasts of the skin) or human multi-potent adult stem cells (i.e. mesenchymal stem cells) to become even embryonic-like stem cells. Next to this purely scientific scenario, it opened also a complex regulatory and economic context. In the field of a cell therapy, stem cells are configured legally as drugs, and as such, they must therefore be isolated, characterized, expanded and cryopreserved in a strictly manner, codified at international level (the so-called “GMP”). From the economic point of view, the impact of Regenerative Medicine is crucial to the sustainability of public health costs in an aging population. The turnover of cell-based products is growing at a rate of 10-15% /year and is expected to reach $ 6.6 M in 2016.

The INBB is active in the field, in particular in the INBB Laboratory of Molecular Biology and Stem Cell Bioengineering at the University of Bologna, with the creation of innovative strategies for regenerative medicine of cardiovascular damage. As a result of the activities of the INBB Laboratory were identified natural molecules that can act as directors of cardio-genesis in stem cells and new molecules have been synthesized with a “logical differentiation and paracrine”. Cocktails of natural molecules were developed in order to increase the paracrine activities of human mesenchymal stem cells, also to support the survival and functionality of pancreatic islets in vivo. It was produced the first evidence in the literature of the ability of magnetic fields to modulate the degree of multi-potency of stem cells, together with their ability to differentiate into multiple phenotypes.
It was produced the first evidence in the literature of the possibility of reprogramming, with electromagnetic fields, human adult somatic cardiac non-stem, neuronal and skeletal muscle, without the use of gene transfer with viral vectors, and without altering the sense pro-oncogenic the cellular homeostasis. The discovery of the properties of the cells to emit detectable and audible nano-mechanical vibrations as sound waves has opened up unprecedented prospectives to the study of the health of cells and modulation of cellular differentiation destinations.
In an attempt to develop systems of cell therapy rapidly transferable in the clinical environment, it has been realized for the first time a method and a device capable of isolating a fraction from human adipose tissue micronized containing a niche vasculo-stromal intact and, in its interior, cells mesenchymal stem cells and pericytes. This form of tissue, unlike the lipo-aspirate, it is cryo-storable, also from cadaver donor, without any loss of architecture of the niche vasculo-stromal and vitality of its cellular elements. The commitment of the INBB researchers in this specific area is also reported from the activity reports submitted as part of the INBB National Conferences, among which are mentioned the most recent ones related to INBB IX National Conference, held in Rome on 21-22 October 2010 and X INBB National Conference, held in Rome on 22-23 October 2012, whose detailed programs and abstracts of the reports on the website www.inbb.it

The research lines on which INBB researchers are working and will address in the near future their activities are illustrated as follows:

• Identification of natural molecules responsible for cardio-genesis. These studies have led to the identification of endorphins as directors of the natural process of cardio-genesis in the stem cells. The research have also identified, for the first time, the presence of nuclear receptors and signal intra-nuclear transduction pathways for these molecules.
• Synthesis of innovative molecules with a differentiation logic and paracrine. For the first time, it has been synthesized mixed esters of hyaluronic acid, butyric acid and retinoic acid (HBR), capable of acting with epigenetic modulation and re-modelling of the state of histone acetylation, inducing a significant increase in myocardial and vascular targeted differentiation of human mesenchymal stem cells from different sources (bone marrow, dental pulp, placenta at term). Overall, these studies open the way to a chemical rescue, immediate and lasting of the infarcted myocardium, allowing to avoid the formation of fibrous scar tissue and at the same time, to secure the cardiac tissue for the time needed to initiate therapy cell with stem cell transplant autologous or allogeneic.
• Use of physical energy for regenerative medicine. These studies demonstrated for the first time in the literature the possibility of using extremely low frequency electro-magnetic fields or radio-electric fields channelled asymmetrically to modulate the multi-potency of stem cells, increase the differentiation potential and paracrine stem, reprogram adult somatic cells not-stem elements in pluripotent and fight the aging of the stem cell population. We have also demonstrated for the first time the emission of audible vibrations by cells, opening the way to the use of sound as a “vibrational signature” of the processes of cell transformation and useful information to guide the stem cell and somatic differentiation and reprogramming.
• Development of new methods and devices for the isolation of mesenchymal tissues cryo-storable containing elements and pericytes within an intact niche stromal-vascular. These research have developed a system and a method based on the use of weak mechanical forces for the isolation of a fraction of micronized fat from human lipoaspirate. The resulting product (Lipogems) was found to be cryo-storable, also if it is from deceased donor. From the biological point of view it is a tissue which contains intact inside the micro-environment in which stem cells live. From the regulatory point of view, the product Lipogems is a derivative of the adipose tissue with the characteristics of a product that can be easily manipulated minimally injected to autologous donor. The general procedure is very fast and safe, it does not require the expansion or manipulation of stem cells, and therefore is not subject to any restrictions imposed by the standards of good manufacturing practice.

In addition to INBB National Laboratory of Molecular Biology and Stem Cell Bioengineering active at the University of Bologna, different RUs operate on INBB Platform on “Regenerative Medicine and Stem Cells”: first the RU of the University of Naples Federico II and Pavia.

SOME RECENT PUBLICATIONS

• Cavallari G, Olivi E, Bianchi F, Neri F, Foroni L, Valente S, La Manna G, Nardo B, Stefoni S, Ventura C. Mesenchymal stem cells and islet cotransplantation in diabetic rats: improved islet graft revascularization and function by human adipose tissue-derived stem cells preconditioned with natural molecules. Cell. Transplant. 21:2771-2781, 2012
• Cantoni S, Galletti M, Zambelli F, Valente S, Ponti F, Tassinari R, Pasquinelli G, Galiè N, Ventura C. Sodium butyrate inhibits platelet-derived growth factor-induced proliferation and migration in pulmonary artery smooth muscle cells through Akt inhibition. FEBS J. 280:2042-2055, 2013.
• Cantoni S, Cavallini C, Bianchi F, Bonavita F, Vaccari V, Olivi E, Frascari I, Tassinari R, Valente S, Lionetti V, Ventura C. Rosuvastatin elicits KDR-dependent vasculogenic response of human placental stem cells through PI3K/AKT pathway. Pharmacol. Res. 65:275-284, 2012
• Lionetti V, Ventura C. Regenerative medicine approach to repair the failing heart. Vascul. Pharmacol. 58:159-163, 2013
• Maioli M, Rinaldi S, Santaniello S, Castagna A, Pigliaru G, Gualini S, Fontani V, Ventura C. Radiofrequency energy loop primes cardiac, neuronal, and skeletal muscle differentiation in mouse embryonic stem cells: a new tool for improving tissue regeneration. Cell Transplant.; 21:1225-1233, 2012
• Rinaldi S, Maioli M, Santaniello S, Castagna A, Pigliaru G, Gualini S, Margotti ML, Carta A, Fontani V, Ventura C. Regenerative treatment using a radioelectric asymmetric conveyor as a novel tool in antiaging medicine: an in vitro beta-galactosidase study. Clin. Interv. Aging 7:191-194, 2012
• Maioli M, Rinaldi S, Santaniello S, Castagna A, Pigliaru G, Gualini S, Cavallini C, Fontani V, Ventura C. Radio electric conveyed fields directly reprogram human dermal skin fibroblasts toward cardiac, neuronal, and skeletal muscle-like lineages. Cell Transplant. 22:1227-1235, 2013
• Collodel G, Fioravanti A, Pascarelli NA, Lamboglia A, Fontani V, Maioli M, Santaniello S, Pigliaru G, Castagna A, Moretti E, Iacoponi F, Rinaldi S, Ventura C. Effects of regenerative radioelectric asymmetric conveyer treatment on human normal and osteoarthritic chondrocytes exposed to IL-1β. A biochemical and morphological study. Clin. Interv. Aging 8:309-316, 2013
• Maioli M, Rinaldi S, Santaniello S, Castagna A, Pigliaru G, Delitala A, Lotti Margotti M, Bagella L, Fontani V, Ventura C. Anti-senescence efficacy of radio-electric asymmetric conveyer technology. Age (Dordr). 2013 May 9. [Epub ahead of print]
• Bianchi F, Maioli M, Leonardi E, Olivi E, Pasquinelli G, Valente S, Mendez AJ, Ricordi C, Raffaini M, Tremolada C, Ventura C. A new non-enzymatic method and device to obtain a fat tissue derivative highly enriched in pericyte-like elements by mild mechanical forces from human lipoaspirates. Cell Transplant. 2013;22(11):2063-77