Horses and HBOT

Equine Vet J. 2012 Feb 15. doi: 10.1111/j.2042-3306.2011.00536.x. [Epub ahead of print]

(photo from http://www.sanctuaryequinerehab.com)

Equine peripheral blood-derived mesenchymal stem cells: Isolation, identification, trilineage differentiation and effect of hyperbaric oxygen treatment.

Dhar M, Neilsen N, Beatty K, Eaker S, Adair H, Geiser D.

Source

Departments of Large Animal Clinical Sciences Pathobiology, College of Veterinary Medicine, University of Tennessee, Knoxville, USA GE Healthcare Cell Therapy Division, Piscataway, New Jersey, USA.

Abstract
Reasons for performing study: Two studies report variability in proliferation and limited adipocyte differentiation of equine peripheral blood-derived adult mesenchymal stem cells, thus casting doubt on their adipogenic potential. Peripheral blood can be a valuable source of adult mesenchymal stem cells if cell culture conditions permissive for their adherence, proliferation and differentiation are defined. Hyperbaric oxygen treatment has been reported to mobilise haematopoietic progenitor stem cells into the peripheral blood in humans and mice, but similar experiments have not been done in horses. Objectives: To optimise cell culture conditions for isolation, propagation and differentiation of adult stem cells from peripheral blood and to assess the effect of hyperbaric oxygen treatment on adult stem cell concentrations. Methods: Peripheral blood was collected from the jugular vein of 6 research mares, and mononuclear cells were isolated. They were subjected to cell culture conditions that promote the adherence and proliferation of adult stem cells. The cells were characterised by their adherence, expression of cellular antigen markers, and trans-differentiation. Each horse was subjected to 3 hyperbaric oxygen treatments, and stem cells were compared before and after treatments. Stem cells derived from adipose tissue were used as controls. Results: One-third of the horses yielded viable stem cells from peripheral blood, positive for CD51, CD90 and CD105, and demonstrated osteocyte, chondrocyte and adipocyte differentiation. Hyperbaric oxygen treatment resulted in a significant increase in CD90-positive cells. Horses that did not yield any cells pretreatment did so only after 3 hyperbaric oxygen treatments. Conclusions and potential relevance: Peripheral blood can be a valuable source of adult stem cells, if one can identify reliable equine-specific markers, provide methods to increase the number of circulating progenitor cells and optimise cell culture conditions for growth and viability. Our findings are important for further studies towards technological advances in basic and clinical equine regenerative medicine.

Hyperbaric Oxygenation Increases Patients own Stem Cells By Eight-Fold

Hyperbaric Oxygenation Increases Patients own Stem Cells By Eight-Fold

... 2 hours HBOT at 2 ATA; doubles the patients own circulating stem cells
... 40-60 hours HBOT increases circulating stem cells by 8-fold (800%) !!
A scientific study completed at the University of Pennsylvania School of Medicine reports that Hyperbaric Oxygen Therapy (HBOT) are a safe and effective way to mobilize the patients own stem cells providing immediate benefit and further preparing the patient for future stem cell implantation related therapies.
In fact the population of CD34+ cells in the peripheral circulation of humans doubled in response to a single exposure to 2.0 atmospheres absolute (ATA) HBOT for 2 hours. Over a course of twenty treatments, circulating CD34+ cells increased eight-fold!
Stem cells, also called progenitor cells, are crucial to the repair of injured tissues and organs. Hyperbaric Oxygenation increases by eight-fold the number of circulating stem cells throughout the body. Healthy recovery of injured and diseased tissues is the ultimate goal and stem cells play an essential role.
In response to injury, stem cells are mobilized out of the bone marrow to the injured sites, where they differentiate into specialized cells that are important to the healing process. Stem cells from bone marrow are capable of providing specialized functions in many different organs and tissues throughout the body. This movement, or mobilization, of stem cells can be triggered by a variety of stimuli—including Hyperbaric Oxygenation.
While drugs are associated with a host of side effects, Hyperbaric Oxygenation treatments carry a significantly lower risk of such effects.
"This is the safest way clinically to increase stem cell circulation, far safer than any of the pharmaceutical options," said Stephen Thom, MD, Ph.D., Professor at the University of Pennsylvania School of Medicine and lead author of the study.
"This study provides information on the fundamental mechanisms for hyperbaric oxygen therapy and offers a new therapeutic option for mobilizing stem cells."
"We reproduced the observations from humans in animals in order to identify the mechanism for the hyperbaric oxygen effect," added Thom. "We found that hyperbaric oxygen mobilizes stem/progenitor cells because it increases synthesis of a molecule called nitric oxide in the bone marrow. This synthesis is thought to trigger enzymes that mediate stem/progenitor cell release."
Hyperbaric Oxygenation not only causes the release of the patients circulating stem cells but greatly facilitates future endeavors using stem cell related therapies which is costly and not an automatic guarantee in every patient.
It is hoped that future study of hyperbaric oxygen's role in mobilizing stem cells will provide a wide array of treatments for combating injury and chronic progressive disease.
The completed study is scheduled for publication in the April 2006 edition of the American Journal of Physiology – Heart and Circulatory Physiology.
Submitted on August 19, 2005; Accepted on November 7, 2005
Stem cell mobilization by hyperbaric oxygenation
Stephen R Thom1, Veena M Bhopale2, Omaida C Velazquez3, Lee J Goldstein3, Lynne H Thom2*, and Donald G Buerk4
1 Emergency Medicine, University of Pennsylvania, Philadelphia, PA, USA; Institute for Environmental Medicine, University of Pennsylvania, Philadelphia, PA, USA 2 Emergency Medicine, University of Pennsylvania, Philadelphia, PA, USA 3 Surgery, University of Pennsylvania, Philadelphia, PA, USA 4 Physiology, University of Pennsylvania, Philadelphia, PA, USA We hypothesized that exposure to hyperbaric oxygen (HBO2) would mobilize stem/progenitor cells from the bone marrow by a nitric oxide (.NO) dependent mechanism. The population of CD34+ cells in the peripheral circulation of humans doubled in response to a single exposure to 2.0 atmospheres absolute (ATA) O2 for 2 hours. Over a course of twenty treatments, circulating CD34+ cells increased eight-fold, although the over-all circulating white cell count was not significantly increased. The number of colony-forming cells (CFCs) increased from 16 ± 2 to 26 ± 3 CFCs/100,000 monocytes plated. Elevations in CFCs were entirely due to the CD34+ sub-population, but increased cell growth only occurred in samples obtained immediately post-treatment. A high proportion of progeny cells express receptors for vascular endothelial growth factor-2 and for stromal derived growth factor. In mice, HBO2 increased circulating stem cell factor by 50%, increased the number of circulating cells expressing stem cell antigen-1 and CD34 by 3.4-fold, and doubled the number of CFCs. Bone marrow .NO concentration increased by 1008 ± 255 nM in association with HBO2. Stem cell mobilization did not occur in knock out mice lacking genes for endothelial .NO synthase. Moreover, pre-treatment of wild type mice with a nitric oxide (.NO) synthase inhibitor prevented the HBO2-induced elevation in stem cell factor and circulating stem cells. We conclude that HBO2 mobilizes stem/progenitor cells by stimulating .NO synthesis.