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Endothelial Cell Mediated Delay of Blood Brain Barrier Recovery Following Tra...Arthur Stem
TBI is the leading cause of death among young adults and children in the developed world, accounting for over 50,000 deaths per year. [12] TBI results in a sleuth of poor health outcomes, including hemorrhaging, seizures, neural edema, neural inflammation, and cognitive and emotional disabilities. All of these outcomes are a direct result of fundamental degradation of the BBB over a time course post TBI. [1] [12] The BBB is an integral structure that forms around the microvascular of the cerebral cavity. Endothelial cells form the basal membrane through which strictly controlled movement of molecules is observed between the extravascular and intravascular space across this basal membrane. This basal membrane is maintained by endothelial cells, having tight junctions between them to make up the pores through which transport of molecules can occur between the brain and microvasculature. These tight junctions are maintained through cross-talk between the endothelial cells and supporting neurons such as astrocytes and pericytes. [2] A multitude of proteins make up the tight junctions between the endothelial cells, including six main scaffolding structures Claudins 1, 3, and 5, ZO-1, Occludins, and Cadherins. [3] VEGF release following trauma induces endothelial cells to release matrix metalloproteinases (MMPs), in particular MMP9, which can catalyze the N-terminal amino acids that compose the tight junction protein ZO-1. [10] [11] MMP9 when in circulation is also known to activate tumor necrosis factor alpha (TNFɑ) which in turn upregulates transcription of MMP9, creating a positive feedback loop. [11] The management of MMP production is three fold, transcription, proenzyme activation, and substrate inhibition. [11] In our study, it is proenzyme activation via TP that is the focus and how that affects the overall transcription levels of the tight junction proteins within the endothelial cells and astrocytes.
Dr. Hughes discusses the TBI Therapy mutlimodal approach to treating traumatic brain injury including hyperbaric oxygen therapy (HBOT), IV nutrition, intransal PRP, insulin, and adult stem cells, cranial osteopathy, and the ketogenic diet at the OMED conference in Baltimore.
Dr. David Steenblock has been specializing in regenerative medicine for over 40 years. This power point discusses how stem cells can regenerate the body and help you heal. To learn more about stem cell treatments, call 1-800-300-1063.
Pathophysiology of TBI is complex and consists of acute and delayed injury. In the acute phase, brain tissue destroyed upon impact includes neurons, glia, and endothelial cells, the latter of which makes up the blood-brain barrier. In the delayed phase, “toxins” released from damaged cells set off cascades in neighboring cells eventually leading to exacerbation of primary injury. As researches further explore pathophysiology and molecular mechanisms underlying this debilitating condition, numerous potential therapeutic strategies, especially those involving stem cells, are emerging to improve recovery and possibly reverse damage. In addition to elucidating the most recent advances in the understanding of TBI pathophysiology, this review explores two primary pathways currently under investigation and are thought to yield the most viable therapeutic approach for treatment of TBI: manipulation of endogenous neural cell response and administration of exogenous stem cell therapy.
A Multimodal Regenerative Approach to TBI - ROME 2019Megan Hughes
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A Multimodal, Regenerative Approach to Traumatic Brain Injury
1. A Multimodal, Regenerative Approach to
Traumatic Brain Injury
John C. Hughes, D.O.
Academy of Regenerative Practices
Conference and Scientific Seminar
Fort Lauderdale, FL
February 24th-26th, 2017
2. DISCLAIMER
• I have no relevant financial relationships with
any commercial interests to disclose.
• The content of this presentation has been peer reviewed
for fair balance and evidence based medicine.
4. MAINSTREAM TREATMENTS
• Occupational and physical rehabilitation, speech
therapy, pharmaceutical drugs, and cognitive
maintenance exercises.
• Patients resign to simply cope with their condition as
they reach a plateau of overall treatment benefit.
5. ALTERNATIVE
TREATMENTS
• Do not seek to regenerate the
damaged brain function TBI but
rather simply treat symptoms
(aka hormone replacement
therapy).
• Do not combine regenerative
treatments in a multimodal
manner in order to maximize
patient benefit.
• Singular treatments can be
prohibitive for patients and their
families, both in cost and time.
6. REGENERATIVE, MULTIMODAL
APPROACH TO TBI
I. Hyperbaric Oxygen Therapy
II. Intranasal Therapies
III. IV Nutrition
IV. Cranial Osteopathy
V. Ketogenic Diet and MCT Oil
7. TBI THERAPY:
THESIS
It is hypothesized that the practical, effective combination of
multiple regenerative TBI therapies can produce synergistic benefits
to the patient that exceed the use of one particular TBI treatment.
9. TBI THERAPY:
HYPERBARIC OXYGEN THERAPY
• Breathing 100% oxygen at increased
atmospheric pressures allows the body
to absorb about 10-15 times its normal
supply of oxygen.
• This “high dose” oxygen, stimulates
the growth of tissue, bone and blood
vessels, reduces inflammation, and
mobilizes stem cells.
10. Volume rendered Brain SPECT perfusion maps of Example 1, a 51-year-old woman from the
treated group suffering mTBI that had occurred 2 years prior to inclusion in the study.
Boussi-Gross R, Golan H, Fishlev G, Bechor Y, Volkov O, et al. (2013) Hyperbaric Oxygen Therapy Can Improve Post Concussion Syndrome Years after Mild Traumatic Brain Injury -
Randomized Prospective Trial. PLoS ONE 8(11): e79995. doi:10.1371/journal.pone.0079995
11. TBI THERAPY:
HYPERBARIC OXYGEN THERAPY
• 2 hours of HBOT at 2 ATA triples the patients own
circulating stem cells (SPCs) and doubles CD34+ cells in
the peripheral circulation (Thom, 2005).
• 20 sessions of HBOT at 2 ATA increases circulating
CD34+ stem cells to 8 fold (800%) (Thom, 2005).
• This stem cell increase results from an increase in nitric
oxide in the bone marrow. Nitric oxide stimulates
enzymes that mediate stem/progenitor cell release.
12. Mean CD34+ population in blood of humans before and after HBO2 treatments. Data are the
fraction of CD34+ cells within the gated population using leukocytes obtained from 26
patients before and after their 1st, 10th, and 20th HBO2 treatment. *Repeated-measures one-
way ANOVA, P < 0.05 vs. the pre-HBO2 first treatment value.
Thom, S. R., Bhopale, V. M., Velazquez, O. C., Goldstein, L. J., Thom, L. H., & Buerk, D. G. (2006). Stem cell mobilization by hyperbaric oxygen. American Journal of Physiology-
Heart and Circulatory Physiology, 290(4), H1378-H1386.
13. “
”Stephen Thom, MD, Ph.D. (2005)
[Hyperbaric oxygen therapy] is the safest way
clinically to increase stem cell circulation, far
safer than any of the pharmaceutical options.
15. Danielyan, L., Beer-Hammer, S., Stolzing, A., Schäfer, R., Siegel, G., Fabian, C., ... & Novakovic, A. (2014). Intranasal delivery of bone marrow-derived mesenchymal stem cells, macrophages,
and microglia to the brain in mouse models of Alzheimer’s and Parkinson’s disease. Cell transplantation,23(1),S123-S139.
Schematic drawing of two routes of IN delivery of cells to the brain. After crossing the cribriform plate
(CP), the olfactory route (OR, red arrows) divides into two branches: (1) the CSF branch and (2) the
parenchymal branch. Solid arrows represent the paths of migration of cells into the brain evidenced in this
study, whereas dashed arrows reflect possible hypothetical routes of cell delivery. The hypothetical
trigeminal route (TR) consists also of at least two branches one of which crosses the cribriform plate into
the parenchyma, where it diverges to the rostral and caudal parts of the brain. The second branch projects
from the nasal mucosa to the trigeminal ganglion, where the exogenously applied cells are further
distributed to the forebrain, OB and caudal brain areas including the brainstem and the cerebellum.
16. TBI THERAPY:
INTRANASAL THERAPIES-
INSULIN
• Improves brain ATP production
• Decreases CSF cortisol
• Improves neuronal viability in the hippocampus
• Increases the expression of anti-inflammatory microglia
• Reduces beta-amyloid and tau protein deposition
17. NeuN, an immunohistochemical marker of neurons, was used to examine the effect of intranasal
insulin on neurons after injury. Qualitative assessment of histology showed improved neuronal
viability in the hippocampus of the insulin treated rats.
Brabazon, F. P., Khayrullina, G. I., Frey, W. H., & Byrnes, K. R. (2014, June). INTRANASAL INSULIN TREATMENT OF TRAUMATIC BRAIN INJURY. In JOURNAL OF
NEUROTRAUMA (Vol. 31, No. 12, pp. A106-A106). 140 HUGUENOT STREET, 3RD FL, NEW ROCHELLE, NY 10801 USA: MARY ANN LIEBERT, INC.
Intranasal insulin increases the expression of anti-inflammatory microglia in the hippocampus.
18. TBI THERAPY:
INTRANASAL THERAPIES-
PLATELET RICH PLASMA (PRP)
Autologous plasma contains growth
factors and cytokines to aid the injured
brain:
• VEGF, EGF increases angiogenesis
• PDGF, TGF-p enhance collagen growth
• IGF-1 stimulates protein synthesis
Enhanced collagen IV in neurons of the brain has been shown to have a
neuroprotective effect and reduce amyloid-beta proteins.
19. TBI THERAPY:
INTRANASAL THERAPIES-
PLATELET RICH PLASMA (PRP)
• The infusion of concentrated platelets, once activated,
results in an exponential increase in numerous growth
factors at the sight of infusion
• Plasma cytokines:
• Control inflammatory mediators cox1, cox2
• Guide stem cells to areas of injury
20. TBI THERAPY:
INTRANASAL THERAPIES-
PLATELET RICH PLASMA (PRP)
• “Basic fibroblast growth factor (bFGF) or epidermal growth factor
(EGF) infusion enhances injury-induced cell proliferation in the
dentate gyrus (DG) and improves cognitive function in rats
following fluid percussive injury” (Sun, 2014).
• “Other studies have found that infusion of S100β or VEGF can also
enhance neurogenesis in the hippocampus and improve the
functional recovery of animals following TBI” (Kleindienst et al.,
2005; Lee and Agoston, 2010; Thau-Zuchman et al., 2010 cited from
Sun, 2014).
21. TBI THERAPY:
INTRANASAL THERAPIES-
STEM CELLS
• Peripheral blood derived
pluripotent stem cells that are
released from the bone
marrow are used.
• Plasma contains hundreds of
thousands of these cells per
ml in peripheral blood
depending on age.
22. TBI THERAPY:
INTRANASAL THERAPIES-
STEM CELLS
• IN adult stem cells have been used to treat ischemic brain
damage by reducing gray and white matter loss.
• IN adult stem cells downregulate neuroinflammatory
cytokines and rescue tyrosine hydroxylase (TH)-positive
neurons in the substantia nigra in AD and PD patients.
23. TBI THERAPY:
INTRANASAL THERAPIES-
NUTRIENTS
• IN glutathione has been used to reduce oxidative
stress and enhance cellular detoxificaton in
Parkinson’s disease patients.
• IN methylcobalamin has been shown to improve
QEEG Theta activity in ADHD and autism
patients.
25. TBI THERAPY:
INTRAVENOUS NUTRITION
• Vitamin D deficiency has been found in over 65% of TBI
patients suffering from chronic fatigue.
• B vitamin supplementation (particularly B12, folate, B6) has
been found to improve memory, mood, and energy levels and
has been used to prevent stroke and Alzeheimer’s disease.
27. TBI THERAPY:
CRANIAL OSTEOPATHY
• The central nervous system, including the brain and
spinal cord, has a subtle, rhythmic pulsation which can
be manipulated by a skilled practitioner.
• This rhythmic pulsation (aka CRI) can be blocked in
brain injuries in which CSF and blood flow is impeded
and the brain loses nourishment.
• Cranial osteopathy has been found effective at treating
the vertigo and headaches associated with TBIs.
28. Time shift between peaks of TCD and В-Imp is determined by the replacement of some portion
of CSF out from (or into) zone of В-Imp electrodes. This time interval represents the mobility оf
CSF inside the cranium during the pulse cycle. At this period no active processes could operate.
Investigations under different conditions have shown that "t" reflects CSF mobility.
Moskalenko, Y., Frymann, V., Kravchenko, T., & Weinstein, G. (2003). Physiological background of the Cranial Rhythmic Impulse and the Primary respiratory Mechanism. Am Acad Osteopath
J, 13(2), 21-33.
30. TBI THERAPY:
KETOGENIC DIET
• Proven treatment for patients suffering from
epilectic seizures.
• Ketogenic diets studied in brain trauma (CCI)
produce corticol sparing and less apoptotic
neuro-degeneration and overall improvements in
cognitive and motor functioning.
31. TBI THERAPY:
MCT OILS
• Increase the available calming neurotransmitter
GABA, thereby reducing neuronal
hyperpolarization and the excititatory
neurotransmitter glutamate.
• With less glutamate, there is less oxidative stress and
improved neuroprotection.
• MCT oils are a rich source of ketone bodies.
32. McNally, M. A., & Hartman, A. L. (2012). Ketone bodies in epilepsy. Journal of neurochemistry, 121(1), 28-35.
Possible anticonvulsant effects of
ketone bodies on the brain.
(1) Increased GABA synthesis
through alteration of glutamate
cycling in glutamate-glutamine
cycle or altered neuronal
responsiveness to GABA at
GABAA receptors.
(2) Decreased glutamate release by
competitive inhibition of
vesicular glutamate transporters.
(3) Other neurotransmitters,
including norepinephrine and
adenosine.
(4) Increased membrane potential
hyperpolarization via
KATP channels possibly mediated
by GABAB receptor signaling.
(5) Decreased reactive oxygen
species production from
glutamate exposure.
(6) Electron transport chain subunit
transcription.
Abbreviations: A1R, adenosine receptor; Cl, chloride; GLN, glutamine; GLU, glutamate; GABA, γ-aminobutyric acid ;
GABABR, γ-aminobutyric acid beta receptor; GABAAR, γ-aminobutyric acid alpha receptor; VGLUT, vesicular glutamate
transporter; ROS, reactive oxygen species.
34. TBI THERAPY:
PROTOCOL
• HBOT: at 1.3 ATA to 1.75 ATA from 10 to 40 sessions
• Intranasal therapies: utilized 1 to 4 x during HBOT treatment
series (IN plasma, insulin, glutathione, B12) administered
first followed by IN platelet-derived, pluripotent stem cells
within 7 days of IN plasma
• patients are also sent home with 10 days IN insulin to self administer
• Cranial osteopathy: administered throughout HBOT
treatment series
35. TBI THERAPY:
PROTOCOL
• IV nutrition: administered 1-4 x during HBOT treatment
series
•Myer’s cocktail with potassium, magnesium, calcium, B-complex,
B5, B6, and B12, ascorbate followed by a glutathione push
• Ketogenic Diet, MCT Oils and Supplementation
•Blueberries, Vitamin D3, and elk antler recommended daily 3 weeks
before and after treatment
•Ketogenic dietary counseling and MCT oils are begun on day 1 of
HBOT series and continued for 3 months after treatment
36. TBI THERAPY:
CLINICAL RESULTS
Out of 20 patients treated, every patient has reported:
• More mental clarity
• Improved memory
• Improved executive function/decision making
• More stable emotions and less stress
• Better ability to cope with pain
• More physical and mental energy
37. TBI THERAPY:
CLINICAL RESULTS
Out of 20 patients treated, some patients have reported:
• Less sound and light sensitivity
• Improved eyesight
• Improved sleep and libido
• Improved motor function (ability to open a clenched fist,
ability to walk)
• Less muscle spasticity
38. TBI THERAPY:
CASE REPORT
“I was so hypersensitive to light and sound that I had to
wear ear plugs, headphones, sunglasses, giant sun hat, and
a scarf just to attend the appointment. I could not drive.
After the intranasal PRP treatment, it was like a stream of
information had been let loose like a dam that had busted. I
felt for the first time in a year that I had some clarity. I was
excited and able to read more than 2-3 sentences without
triggering a migraine. I found that I was able to get back
on the computer and learn more about my trauma and
recent treatments. Within the following days it was like an
awakening. It seemed like a light switch was turned back
on inside my head. The ability to think and plan returned.”
39. TBI THERAPY:
CONCLUSION
This regenerative, multi-modal TBI therapy protocol has a
unifying theme of providing regenerative substrates to the
injured brain that correct its metabolic crisis—by increasing
oxygen, nutrients, cofactors, growth factors, and stem cells—
to ultimately repair damaged neurons, enhance neurogenesis,
increase blood flow, improve glucose utilization, decrease
inflammation, and stabilize neurotransmitter release.
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