LOS ANGELES, CA--(Marketwired - Dec 19, 2016) - Endonovo Therapeutics, Inc. (OTCQB: ENDV) ("Endonovo" or the "Company"), a developer of non-invasive electroceuticals for the treatment of vascular diseases and inflammatory conditions, today announced it is commencing a pre-clinical study at a contract research organization to assess the therapeutic potential of its Immunotronics™ platform in the prevention of heart failure following myocardial infarction (MI). The pre-clinical study will evaluate the effect of the Company's non-invasive electroceutical technology on cardiac function, post-MI remodeling, and infarct size, as well as angiogenesis (the development of new blood vessels). The study, estimated to be complete at the end of the first quarter of 2017, represents the first of several planned studies designed to evaluate the Company's proprietary non-invasive electroceuticals in the treatment of vascular diseases and ischemia/reperfusion injury.
"We are leveraging the experience and proprietary knowledge of our scientific team, which has undertaken over 25 years of research and development in the field of bioelectromagnetic therapeutics in angiogenesis, neuroinflammation and ischemia to develop non-invasive treatments for substantially unmet clinical needs," commented Endonovo Chairman and CEO, Alan Collier.
"A positive result in this study would represent a significant milestone for Endonovo and the field of bioelectronic medicine. We are moving to establish Endonovo as the leader in electroceuticals-based regenerative medicine. Our competitors in the bioelectronic medicine space are primarily developing implantable devices targeting the inflammatory response, and our competitors in regenerative medicine are developing cell therapies, biologics and gene therapies to treat many of these diseases, which are more expensive to develop and present significantly more safety concerns for potential patients.
"Our technology presents a lucrative opportunity to develop a non-invasive platform device that can be used to treat cardiovascular, cerebrovascular and peripheral artery disease as well as ischemia/reperfusion injuries," concluded Mr. Collier.
The Company had previously announced receiving a term sheet for $5 million in preferred financing from a strategic healthcare investor to develop a pipeline in vascular diseases, peripheral artery disease and ischemia/reperfusion injury. The $5 million proposed financing is part of a larger $15 million round of financing to uplist the Company's common stock onto a national stock exchange in the first half of 2017.
Physiological and Molecular Genetic Effects of Time-Varying Electromagnetic Fields on Human Neuronal Cells
Thomas J. Goodwin, PH.D., Lyndon B. Johnson Space Center
National Aeronautics and Space Administration (NASA), Johnson Space Center, Houston, Texas
The present investigation details the development of model systems for growing two- and three dimensional human neural progenitor (NHNP) stem cells within a culture medium facilitated by a time-varying electromagnetic field (TVEMF), i.e. PEMF.
The cells and culture medium are contained within a two- or three-dimensional culture vessel, and the electromagnetic field is emitted from an electrode or coil. These studies further provide methods to promote neural tissue regeneration by means of culturing the neural cells in either configuration. Grown in two dimensions, neuronal cells extended longitudinally, forming tissue strands extending axially along and within electrodes comprising electrically conductive channels or guides through which a time-varying electrical current was conducted.
In the three-dimensional aspect, exposure to TVEMF resulted in the development of three-dimensional aggregates, which emulated organized neural tissues.
In both experimental configurations, the proliferation rate of the TVEMF cells was 2.5 to 4.0 times the rate of the non-waveform cells. Each of the experimental setups resulted in similar molecular genetic changes regarding the growth potential of the tissues as measured by gene chip analyses, which measured more than 10,000 human genes simultaneously.
This study clearly shows the ability to use TVEMF to control the proliferative rate, directional attitude, and molecular genetic expression of normal human neural progenitor cells. The procedure is applicable to, but not limited to, the control of NHNP cells in both two-dimensional and three-dimensional culture.
The genetic responses both up-regulated and down-regulated genes which were maturation- and growth-regulatory in nature. These genes are also primarily involved in the embryogenic process.
Therefore it is reasonable to conclude that control over the embryogenic development process may be achieved via the presently demonstrated methodology. Specific genes such as human germline oligomeric matrix protein, prostaglandin endoperoxide synthase 2, early growth response protein 1, and insulin-like growth factor binding protein 3 precursor are highly up-regulated. Keratin Type II cytoskelatal 7, mytotic kinesin like protein 1, transcription factor 6 like 1, mytotic feedback 27 control protein, and cellular retinoic acid binding protein are down-regulated. Each of these two sets is only an example from the approximately 320 genes changes expressed as a consequence of exposure to TVEMF.
There is significant precedent in the literature for the results reported above. Kepler et al. (1990) reported the effects of the neurons with oscillatory properties on the composite of neural networks. This work illustrates the likelihood that a pulse width modulated system might bring on specific responses in neural tissues. As previously discussed, Valentini et al. (1993) demonstrated the ability to enhance the outgrowth of neural fibers on materials that possess a weak electric charge. This would indicate that intense electric fields are not necessarily an essential component of this process, and that a weak and persistent stimulus might yield a measurable effect.
Additional evidence of the effects of magnetic fields exists in the work of Sandyk et al. (1992a). This communication details dramatic improvement of a patient with progressive degenerative multiple sclerosis. Briefly, the patient showed considerable improvement when subjected to treatment at a frequency of 2-7 Hz and an intensity of the magnetic field of 7.5 pico Tesla. These parameters marginally parallel those of this report. In a similar fashion, Sandyk et al. (1992b) reported significant improvement in patients treated with the same field strength and intensity. The ability to suppress or stimulate the growth of non-excitable cells has been reported in mouse lymphoma cells by Lyte et al. (1991). A narrow range of electric field was found to be effective at one end to stimulate and at the other to inhibit the growth of these cells. These data might suggest cellular receptors in all cells. To sustain this notion, Brüstle et al. (1996) reported the potential to use neural progenitors for recapitulation of neural tissues. As would be expected, this would require genetic control at the embryonic level. We believe this study indicates our ability to trigger these parametric events.
As is clearly demonstrated in the human body, the bioelectric, biochemical process of electrical nerve stimulation is a documented reality. The present investigation demonstrates that a similar phenomenon can be potentiated in a synthetic atmosphere, i.e., two-dimensionally or in rotating wall cell culture vessels.
One may use this electrical potentiation for a number of purposes, including developing tissues for transplantation, repairing traumatized tissues, and moderating some neurodegenerative diseases and perhaps controlling the degeneration of tissue as might be effected in a bioelectric stasis field.
The basis for Endonovo’s Time-Varying Electromagnetic Field (TVEMF) technology was created at NASA in conjunction with the development of cell therapies to treat injuries and diseases that astronauts might encounter during long term manned missions in outer space.
The original concept of TVEMF was developed when NASA realized that cellular growth increased as the space shuttle crossed the earth’s magnetic poles. It took over eight years to define and develop a device that produced the same effect. This magnetic field device (TVEMF) was originally designed to enhance cellular expansion (growth) in NASA’s rotating wall bioreactor. After numerous experimental studies, the results were so significant with the bioreactor that a prototype was designed for use directly on the human body.
Moving Beyond the Petri Dish
Cells grown in Petri dishes cannot form complex 3D tissues because they tend to sink within their growth mediums. This posed a limitation on researchers studying how tissues form inside the body.
In the 1970’s, a small group at NASA’s Johnson Space Center (JSC) began to think about outer space as a possible answer. Researchers at JSC theorized that if cells could be grown without the influence of the earth’s gravity, they would not sink to the bottom of the culture container; rather, they would stay suspended in the medium and therefore might assemble and form tissue that more closely resembles tissue in the body.
NASA Bioreactor Simulates Microgravity on Earth
NASA researchers developed a soup can-sized bioreactor that spun on its side, preventing cells from sinking to the bottom by maintaining a continuously free-falling state through the culture media. The bioreactor was able to simulate microgravity on earth, providing researchers with the ability to study how tissues form in the body three-dimensionally. The original goal was to attempt tissue growth on Space Shuttle missions. However, the Challenger disaster in 1986 resulted in the grounding of NASA’s space fleet. Subsequently, NASA researchers switched their efforts to creating a culturing device that simulated some aspects of microgravity on earth.
Experiments in Outer Space
NASA researchers conducted multiple experiments using the bioreactor aboard STS-70, STS-85 and STS-90.
During STS-70 in 1995, researchers tested the performance of the bioreactor in microgravity using colon cancer cells. The cells were able to aggregate and form masses 10 mm in diameter. These masses were 30 times the volume of those grown in the control experiment on the ground.
During STS-85 in 1997, researchers repeated the experiment using colon cancer cells and again, mature, differentiated tissue samples were grown. This confirmed previous results – microgravity is an environment beneficial to cell culture and tissue growth.
During STS-90 in 1998, human renal tubular cells were cultured in the bioreactor for 6 days. Researchers compared the activity of 10,000 genes in the flight and ground cultures and identified several of the control genes for differentiation and three-dimensional tissue formation.
Early on, as NASA researchers analyzed data from their experiments aboard the Space Shuttle, they discovered that cells were growing at an increased rate when passing over the earth’s magnetic poles; suggesting that the earth’s magnetic field was influencing cell growth.
Earth’s Magnetic Field Influences Cell Growth
Additionally, several growth factors and cytokines were upregulated in the bioreactor during cellular expansions.
NASA Researchers Add Pulsed Electromagnetic Fields to Bioreactor
Initially, NASA researchers could not replicate the cell growth achieved aboard the Space Shuttle. NASA was using basic sine waves, which could not produce the same effect.
Over the subsequent eight years, researchers experimented with multiple waveforms, frequencies, and slew rates and found the square waveform was able to help deliver the required results and allowed NASA to identify the exact magnetic field format that increased cell growth 400% and upregulated over 200 growth factors and cytokines (G-CSF, GM-CSF, IL-2) in the rotating wall bioreactor.
It was determined that a square waveform with a low frequency (5-10 Hz) and a very rapid slew rate provided optimized results for cellular expansion.
Shifting Away from Stem Cell Transplants
After achieving astounding results with the TVEMF-bioreactor, NASA researchers began to explore whether the effects of TVEMF on cell growth and growth factor production could be done inside the body.
The basic idea was to create a device that could stimulate the body to mobilize and expand stem cells and then promote their homing to the sites of injury without having to extract stem cells from patients, expand the stem cells in vitro and then re-inject the expanded stem cells into the patient for the treatment.
The goal of the TVEMF device is to mitigate many of the problems that current stem cell treatments face, such as the use of drugs to mobilize stem cells and the use of external growth factors and genetic manipulation for stem cell expansion.
Rabbit Osteotomy Study
This is work conducted at Texas A&M University College of Veterinary Medicine on New Zealand White Rabbits using TVEMF.
An osteotomy was performed on the ulna (forearm) leaving a significant gap that would not heal under normal conditions.
The control group used “sham” TVEMF over the area of the osteotomy and the treatment group used TVEMF stimulation.
The study was not conducted until full healing had occurred, but there were obvious differences between the control group and the treatment group.
The study was actually discontinued because researchers could easily distinguish the difference between the control group and treatment group animals. The treatment group animals were mobile, while the control group animals were immobile.
Photo of the incision: Notice the significant gap in the bone
After 14 days with sham TVEMF stimulation:
No bone growth – notice the gap
After 14 days with TVEMF stimulation:
Already some bone growth
After 28 days with sham TVEMF stimulation:
Still no bone growth – notice the gap
After 28 days with TVEMF stimulation:
Dramatic increase in bone growth
Nude Mouse Study
The “nude mouse” study was performed by the independent firm, Charles Rivers Laboratories Inc. using athymic mice. The lack of a thymus leaves the mice without the ability to mount most types of immune responses.
The purpose of the study was to determine whether a cell line was tumorigenic when injected by the subcutaneous route into athymic nude mice.
TVEMF-expanded stem cells were injected into athymic nude mice. It was determined that the TVEMF-expanded stem cells caused no tumors in the mice within 85 days of the injection of the TVEMF-expanded stem cells.
The study validated that expanded stem cells could be transplanted without harmful effects that have been demonstrated with genetic modification.
The multicolor fluorescence in situ hybridization “mFISH” study was conducted at NASA’s Johnson Space Center to determine whether TVEMF technology caused chromosomal damage to stem cells expanded using the NASA technology.
The mFISH study found that there was no chromosomal damage or change in the expanded adult stem cells 90 days after their expansion.
The presence of chromosomal damage or change could cause a wide array of problems such as tumor formation.
The study verified that stem cells grown in the NASA bioreactor are safer for potential therapies than current methods that use genetic manipulation.
Electromagnetic pulses are responsible for many of the activities in our bodies. They are an integral part of what keeps our hearts beating and brains functioning.
The bioelectronics device is a portable Time-Varying Electromagnetic Field (TVEMF) system. The unit uses a sophisticated miniaturized circuit board that delivers the specifically designed and patented electromagnetic field to a designated area of the body through the use of antennas (coils).
TVEMF vs PEMF
Time-Varying Electromagnetic Field (TVEMF) technology is fundamentally different from other magnetic pulse devices. TVEMF is based on the physics of electricity and magnetism as well as the physiology of cells and tissues.
- PEMF devices do not produce square waves, the optimal waveform for a tissue response.
PEMF devices do not produce enough intensity to stimulate a response; most do not even produce 1 Gauss of intensity.
PEMF devices do not produce a rate of induction (slew rate) that is quick enough to penetrate deep within tissues. TVEMF produces pulses that are only 0.0001 seconds wide.
Furthermore, TVEMF features repeating patterns of pulses that are selected based on known motor-neuron stimulation patterns of musculoskeletal tissues.
Sine waves take too long to reach their peak intensity. The lower slew rate means that they are not effective in producing a cellular/tissue response.
Triangle and sawtooth waves also suffer from a lower slew rate, meaning that they are not effective in producing a cellular/tissue response.
Endonovo’s TVEMF device produces a patented square waveform, which was proven the most effective waveform to increase cell growth and the production of over 200 growth factors and cytokines.
Our Cytotronics™ platform uses Time-Varying Electromagnetic Field (TVEMF) technology to expand and enhance the therapeutic properties of stem cells, immune cells and for use in tissue engineering. The goal of our Cytotronics™ platform is to create optimized cell-based therapies with greater therapeutic potential than the un-modulated cells currently being used in regenerative medicine.
The origins of our Cytotronics™ platform dates back to experiments conducted at NASA to expand stem cells ex vivo and to create therapies that could be used to treat astronauts during long term space exploration. The results of those experiments revealed that Time-Varying Electromagnetic Fields (TVEMF) could be used to expand stem cells in the lab and resulted in the increased expression of dozens of genes related to cell growth, tumor suppression, cell adhesion and extracellular matrix production.
A Physics Approach to Biology – Ex Vivo Modulation
We have built upon over 15 years of research and development to create a novel therapeutic paradigm – the systematic enhancement of the biological and therapeutic properties of cells ex vivo. This allows for the creation of a precise, scalable and cost effective approach to maximize the safety and effectiveness of cell-based therapies.
While our competitors are using genetic modification and pharmacological modulation to alter and enhance the biological properties of cells, we are taking A Physics Approach to Biology™ in the use of bioelectronics to create ex vivo modulated cells with enhanced biological and therapeutic properties. Using our Cytotronics™ platform, we are able to create cells that express higher levels of key genes related to stem cell maintenance, cell growth, cell homing and engraftment. Cytotronic™ expansion of peripheral blood stem cells resulted in an over 80-fold expansion of CD34+ cells in as few as 6 days.
Our Cytotronic™ platform was shown to be able to maintain normal human neural progenitor (NHNP) cells in long term culture (180 days), undifferentiated with the ability to be cryogenically preserved then subsequently reintroduced into culture with stem and progenitor markers still intact. NHNP cells that were expanded using our technology were then shown to not be tumorigenic when introduced into nude (athymic) mice for 85 days. Multi-color Fluorescence In Situ Hybridization (mFISH) showed that there was no chromosomal damage to cells subjected to Time-Varying Electromagnetic Fields (TVEMF).
Perivascular Cell Therapy Created Using Cytotronics™
Each year, an estimated 60,000 patients with leukemia and lymphoma need bone marrow transplantation. However, only 25,000 of those patients actually receive a transplant, primarily because the other 35,000 patients are unable to find a fully matched bone marrow donor.
Umbilical cord blood from newborn children is an excellent source of hematopoietic stem cells for stem cell transplants because their immune system is still immature and the stem cells have a lower probability of inducing an adverse immune response in patients. Furthermore, a perfect immunological match between donor and recipient is not necessary, unlike in bone marrow transplants.
However, in most cases a unit of umbilical cord blood contains too few stem cells to treat an adult patient and its use is confined above all to the treatment of children. Numerous patients have better matched cord blood products available (5/6 or 6/6 HLA match), however those products have low cell doses and are not suitable for use in adult patients. This is one of the reasons why less than 3 percent of cord blood collected in the United States is ever used. Low volume cord blood units present the opportunity to expand and/or enhance these unused cord blood units for the widespread use of hematopoietic stem cells in regenerative medicine.
Our Perivascular Cell Therapy is created using immunologically privileged and immune-modulating stem cells from a portion of the human umbilical cord and co-cultured with adipose-derived stem cells along with cord blood cells to create a perivascular cell mixture that can be used to treat malignant and non-malignant hematological disorders.
Endonovo’s cell therapy is an off-the-shelf therapeutic that can be stored indefinitely in a low temperature freeze without requiring cryopreservation. Our technologies are particularly suited to expand the use and effectiveness of low-volume cord blood units.
More In Vivo-Like Stem Cells
Our off-the-shelf therapeutic is created in a three-dimensional bioreactor using our proprietary Cytotronics™ platform to further expand and enhance the biological properties of the stem cells within the perivascular cell mixture. Our use of a perivascular co-culture in a three-dimensional bioreactor is meant to mimic the way that blood-forming stem cells renew and reside in the body. This simulated stem cell niche allows for the expansion of long-term self-renewing stem cells. Researchers had previously identified endothelial and perivascular cells as the cells that were functionally responsible for the maintenance of blood-forming stem cells (HSCs) in the body. Our technologies provide a method for the expansion of HSCs in an environment that resembles their native stem cell niche.
Endonovo is using a three-dimensional bioreactor that allows for the large-scale expansion of blood-forming stem cells that enhances cell-to-cell contact between perivascular cells and HSCs, providing a culture method that more closely resembles the stem cell niche. As seen below, (A) cells grown in three-dimensions become more spherical that those grown in (B) static two-dimensional cultures. Cells grown two dimensionally in Petri dishes or t-flasks sink within their growth medium. These cells do not look or function like real human cells, which grow three-dimensionally in the body.
Alan Collier – Chairman & Chief Executive Officer
Mr. Collier has more than twenty years of experience in corporate finance, IP development, telecommunications and technology with a concentration in healthcare and technology over the last five years. Prior to his appointment as Endonovo’s CEO and chairman, Mr. Collier served as CEO and director of IP Resources International, Inc. where he was instrumental in developing a platform for for the licensing and acquisition of life science and technology companies.
Previously, Mr. Collier served as a Senior Managing Director at Mid-Market Securities, a FINRA-registered Broker-Dealer. Prior to joining Mid-Market Securities, Mr. Collier was a Managing Director at Mosaic capital and co-managed its Capital Markets Group. He was previously a Vice President at Corporate Capital Group and Managing Director and CEO of Greenbridge Capital Group.
Over the progression of his career, he has specialized in the development and financing of early stage, high growth, and acquisitive companies (public and private). He has structured, participated in, and completed numerous transactions including mergers and acquisitions, equity and debt placements, capital restructuring, joint venture development, and channel partner procurement. He has held numerous board and executive positions throughout his career in the telecommunications, technology, specialty finance, corporate finance and healthcare industries. Mr. Collier holds FINRA Series 7, 79, 63, and 24 Licenses.
Michael Scott Mann – President
Mr. Mann has over 30 years experience in merger and acquisitions and operational management. In 2008 Mr. Mann acquired the assets of Hanover Asset Management, now Endonovo Therapeutics, Inc,. and led the company to become listed on the OTCBB in 2012.
From January 2003 to April, 2011, Mr. Mann was the Founder, President and Chief Executive Officer of U.S. Debt Settlement, Inc. (USDS), a Frankfurt listed company, where he implemented a growth by acquisition strategy successfully acquiring numerous companies.
From January 2002 to July 2003, Mr. Mann was the Chief Executive Officer of Shared Vision Capital, a boutique investment banking firm that assisted emerging companies with early seed capital and bridge loans. Earlier still, from October 1998 to December 2001, Mr. Mann was the Vice President of Investor Relations for JuriSearch.com, an online legal research platform. During his tenure with JuriSearch.com, Mr. Mann was directly responsible for funding the company’s growth and development. In addition, Mr. Mann founded Universal Pacific Communications, a privately owned telecommunications company. Under his leadership as President, Universal Pacific developed a fiber optic disaster recovery telecommunications network, designed for and successfully marketed to Fortune 1000 companies.
Leonard Makowka, M.D., Ph.D. – Chief Medical Officer
Over the past 30 years, Dr. Makowka has transitioned himself as a distinguished clinical and transplantation surgeon and medical researcher to a successful entrepreneur, advisor and board member in a wide range of industries. Since receiving his M.D. degree from the University of Toronto Medical School in 1977 and Ph.D. from the University of Toronto’s Department of Pathology in 1982, he has published over 400 articles and chapters in both clinical and basic scientific research.
Between 1989 and 1995, Dr. Makowka served as the Chairman of the Department of Surgery and Director of Transplantation Services at Cedars Sinai Medical Center in Los Angeles, California. During this time, he also served as Professor of Surgery at the UCLA School of Medicine. Dr. Makowka would later become Executive Director of the Comprehensive Liver Disease Center at St. Vincent’s Medical Center in Los Angeles, CA, where he created a multiple disciplinary approach to the treatment of liver disease. He is currently the active Chairman of the Corporate Advisory Board of the UCLA School of Nursing. Dr. Makowka has since retired from the active practice of medicine and has experienced a successful career in developing investment and business strategies for companies in the healthcare, life sciences, finance, and other industries.
Dr. Makowka has served on the board of directors for various private and publicly traded companies in the healthcare and life science industries. From 1998 to 2003 he served on the Board of Directors of Hollis Eden Pharmaceuticals, (NASDAQ: HEPH) and served on the Board of Directors for Kinmed, Inc., a privately held biotech company. As an entrepreneur, Dr. Makowka has worked with several companies in a wide range of industries. He was a founding consultant for Ivivi Technologies, Inc. (NASDAQ: IVVI), a publicly traded medical technology company focusing on designing, developing, and commercializing proprietary electrotherapeutic technologies. In addition, Dr. Makowka founded Trillenium Medical Imaging Inc., a company engaged in the novel healthcare application of proprietary infrared imaging.
As Endonovo Therapeutics’ Chief Medical Officer, Dr. Makowka will be responsible for developing the company’s scientific advisory board, overseeing the company’s clinical research activities and will serve as a liaison between the company and the medical community.
Donnie Rudd, Ph.D., D.Sc., D.D., J.D. – Chief Scientist & Director of Intellectual Property
Dr. Rudd began his career as a Chemical Engineer after graduating with a B. S. degree in Chemical Engineering from Texas A&M. His career then progressed into work as a successful Patent Attorney after obtaining his J.D. degree from Chicago-Kent College of Law. Over the course of 15 years of litigating cases as one of the principals of Rudd & Associates., he was admitted to over 14 courts and taught at 3 universities. Dr. Rudd has written 17 books and over 140 publications.
After a successful tenure and being recognized as “One of the top 5 attorneys in the United States”, he earned his Ph.D. in Bioscience and shifted his focus to stem cell and electromagnetic therapy research.
He joined Regenetech, Inc. in April 2002 as CEO and Corporate Secretary. In 2002 he became its Chief Scientist and Director of Intellectual Property. With patented licensing rights through an innovation from NASA, he lead in the research on the NASA licensed bioreactor. Subsequently, NASA researchers under Dr. Rudd’s guidance added TVEMF technology to the bioreactor. He has developed over 100 different patents and inventions focused on stem cell technology. In accordance with his patents and groundbreaking discoveries in stem cell research, he has become a NASA recognized inventor. For his patent and inventive work during his time at Regenetech, Inc., Dr. Rudd’s work received recognition from the Space Foundation after they inducted the Commercial Earth-Imaging Satellite and Intrifuge CellXpansion technology into the Space Technology Hall of Fame.