|
DPL™ LED Technologies
Deep Penetrating
Light Therapy
NASA Study
|
Medical College of Wisconsin/NASA
NASA
Healthlink
|
 |
NASA Light
Emitting Diode Medical Applications
From Deep Space
to Deep Sea
Harry T. Whelan1a,5,7, Ellen V. Buchmann1a,
Noel T. Whelan1a,7, Scott G. Turner1a,
Vita Cevenini7, Helen Stinson7, Ron Ignatius2,
Todd Martin2,
Joan Cwiklinski1a, Glenn A. Meyer1c, Brian
Hodgson3,4, Lisa Gould1b,
Mary Kane1b, Gina Chen1b, James Caviness6
1aDepartments of Neurology, 1bPlastic
Surgery, 1cNeurosurgery,
Medical College of Wisconsin, Milwaukee, WI
53226, (414) 456-4090
2Quantum Devices, Inc Barneveld, WI 53507
(608) 924-3000
3Children’s Hospital of Wisconsin, Milwaukee,
WI 53201 (414) 266-2044
44th Dental Battalion, 4th Force Service
Support Group, USMCR, Marietta, GA
5Naval Special Warfare Group TWO, Norfolk, VA
23521, (757) 462-7759
6Submarine Squadron ELEVEN, San Diego, CA
92106, (619)553-8719
7NASA-Marshall Space Flight Center, AL 35812,
(256) 544-2121
Abstract:
This work is supported and
managed through the NASA Marshall Space Flight
Center - SBIR Program. LED-technology developed
for NASA plant growth experiments in space shows
promise for delivering light deep into tissues
of the body to promote wound healing and human
tissue growth. We present the results of
LED-treatment of cells grown in culture and the
effects of LEDs on patients' chronic and acute
wounds. LED-technology is also biologically
optimal for photodynamic therapy of cancer and
we discuss our successes using LEDs in
conjunction with light-activated
chemotherapeutic drugs.
We have all heard how space technology can
benefit us all here on earth; well this is no
exception when we look at LED therapy. While the
researchers in the field were fine-tuning their
devices for pain relief, NASA needed a means to
produce light without the added heat produced by
incandescent light bulbs for space missions and
their plant experiments. NASA settled on (LED’s)
because of their ability to produce a scattered
light of various wavelengths that were of
benefit to plants in the confinements of a space
vehicle in space flight, while producing no
significant increase in thermal heat. They
worked, and NASA took the next step. Could LED’s
help in healing injuries to astronauts while in
space flight. One of the major dilemmas for NASA
regarding long-term space flight is the
well-documented effect of muscle and bone
atrophy that occurs to astronauts while in
space. In addition it has been shown that
injuries that occur while in space tend not to
heal until the astronaut is back within the
earth’s gravity. The LED’s that produced
near-infrared light used in NASA’s research were
shown to stimulate the basic energy processes by
activating color sensitive chemicals within the
cells. DNA synthesis in fibroblasts and muscle
cells had been quintupled. The light absorbed by
the cells stimulated the metabolism in muscle
and bone as well as skin and subcutaneous
tissue. What people and animals had felt through
utilizing this technology in real life, NASA was
proving to be true in the laboratory.
LED-ENHANCEMENT
OF CELL GROWTH
Studies on cells exposed to microgravity and
hypergravity indicate that human cells need
gravity to stimulate growth. As the
gravitational force increases or decreases, the
cell function responds in a linear fashion. This
poses significant health risks for astronauts in
long-term space flight. The application of light
therapy with the use of NASA LEDs will
significantly improve the medical care that is
available to astronauts on long-term space
missions. NASA LEDs stimulate the basic energy
processes in the mitochondria (energy
compartments) of each cell, particularly when
near-infrared light is used to activate the
color sensitive chemicals (chromospheres,
cytochrome systems) inside. Optimal LED
wavelengths include 680, 730 and 880 nm and
their laboratory has improved the healing of
wounds in laboratory animals by using both LED
light and hyperbaric oxygen. Furthermore, DNA
synthesis in fibroblasts and muscle cells has
been quintupled using NASA LED light alone, in a
single application combining 680, 730 and 880 nm
each at 4 Joules per centimeter squared.
Muscle and bone atrophy are well documented in
astronauts, and various minor injuries occurring
in space have been reported not to heal until
landing on Earth. An LED blanket device may be
used for the prevention of bone and muscle
atrophy in astronauts. The depth of
near-infrared light penetration into human
tissue has been measured spectroscopically
(Chance, et al., 1988). Spectra taken from the
wrist flexor muscles in the forearm and muscles
in the calf of the leg demonstrate that most of
the light photons at wavelengths between 630-800
nm travel 23 cm through the surface tissue and
muscle between input and exit at the photon
detector. The light is absorbed by mitochondria
where it stimulates energy metabolism in muscle
and bone, as well as skin and subcutaneous
tissue.
Long term space flight, with its many inherent
risks, also raises the possibility of astronauts
being injured performing their required tasks.
The fact that the normal healing process is
negatively affected by microgravity requires
novel approaches to improve wound healing and
tissue growth in space. NASA LED arrays have
already flown on Space Shuttle missions for
studies of plant growth and the U.S. Food and
Drug Administration (FDA) has approved human
trials. The use of light therapy with LEDs can
help prevent bone and muscle atrophy as well as
increase the rate of wound healing in a
microgravity environment, thus reducing the risk
of treatable injuries becoming mission
catastrophes.
Space flight has provided a laboratory for
studying wound healing problems due to
microgravity, which mimic traumatic wound
healing problems here on earth. Improved wound
healing may have multiple applications that
benefit civilian medical care, military
situations and long-term space flight. Laser
light and hyperbaric oxygen have been widely
acclaimed to speed wound healing in ischemic,
hypoxic wounds. An excellent review of recent
human experience with near-infrared light
therapy for wound healing was published by
Conlan, et al (Conlan, 1996). Lasers provide
low energy stimulation of tissues which results
in increased cellular activity during wound
healing (Beauvoit, 1994, 1995; Eggert, 1993;
Karu, 1989; Lubart, 1992, 1997; Salansky, 1998;
Whelan, 1999; Yu, 1997) including increased
fibroblast proliferation, growth factor
synthesis, collagen production and
angiogenesis. Lasers, however, have some
inherent characteristics that make their use in
a clinical setting problematic, such as
limitations in wavelength capabilities and beam
width. The combined wavelengths of light
optimal for wound healing cannot be efficiently
produced, and the size of wounds that may be
treated by lasers is limited. Light-emitting
diodes (LEDs) offer an effective alternative to
lasers. These diodes can be made to produce
multiple wavelengths, and can be arranged in
large, flat arrays allowing treatment of large
wounds. Potential benefits to NASA, military,
and civilian populations include treatment of
serious burns, crush injuries, non-healing
fractures, muscle and bone atrophy, traumatic
ischemic wounds, radiation tissue damage,
compromised skin grafts, and tissue
regeneration.
Combat casualty care in Special Operations
already have adopted the NASA LED technology for
submarines deployed in training with risk of
injury. The USS Salt Lake City is currently
underway with an LED Array in the Pacific.
Special Operations are characterized by lightly
equipped, highly mobile troops entering
situations requiring optimal physical
conditioning at all times. Wounds are an
obvious physical risk during combat operations.
Any simple and lightweight equipment that
promotes wound healing and musculoskeletal
rehabilitation and conditioning has potential
merit. NASA LEDs have proven to stimulate wound
healing at near-infrared wavelengths of 680, 730
and 880 nm in laboratory animals, and have been
approved by the U.S. Food and Drug
Administration (FDA) for human trials. The NASA
LED arrays are light enough and mobile enough to
have already flown on the Space Shuttle numerous
times. LED arrays may be used for improved
wound healing and treatment of problem wounds as
well as speeding the return of deconditioned
personnel to full duty performance. Examples
include: 1. Promotion of the rate of muscle
regeneration after confinement or surgery. 2.
Personnel spending long periods of time aboard
submarines may use LED arrays to combat muscle
atrophy during relative inactivity. 3. LED
arrays may be introduced early to speed wound
healing in the field. Human trials have begun at
the Medical College of Wisconsin, Naval Special
Warfare Command, Submarine Squadron ELEVEN and
NASA-Marshall Space Flight Center.
Wound Healing
with NASA LEDs
EXPERIMENTS USING AN ISCHEMIA ANIMAL MODEL
SYSTEM PROVIDE PRE-CLINICAL DATA RELEVANT TO
HUMAN HEALING PROBLEMS, CHRONIC NON-HEALING
WOUNDS.
LED-Wound Healing
in Rats
An ischemic wound is a wound in which there is a
lack of oxygen to the wound bed due to an
obstruction of arterial blood flow. Tissue
ischemia is a significant cause of impaired
wound healing which renders the wound more
susceptible to infection, leading to chronic,
non-healing wounds. Despite progress in wound
healing research, we still have very little
understanding of what constitutes a chronic
wound, particularly at the molecular level, and
have minimal scientific rationale for
treatment.
In order to study the effects of NASA LED
technology and hyperbaric oxygen therapy (HBO),
we developed a model of an ischemic wound in
normal Sprague Dawley rats. Two parallel 11-cm
incisions were made 2.5 cm apart on the dorsum
of the rats leaving the cranial and caudal ends
intact. The skin was elevated along the length
of the flap and two punch biopsies created the
wounds in the center of the flap. A sheet of
silicone was placed between the skin and the
underlying muscle to act as a barrier to
vascular growth, thus increasing the ischemic
insult to the wounds. The four groups, each
consisting of 15 rats, in this study include:
the control (no LED or HBO), HBO only, LED (880
nm) only, and LED and HBO in combination. The
HBO was supplied at 2.4 atm for 90 minutes, and
the LED was delivered at a fluency of 4J/cm2 for
fourteen consecutive days. A future study will
incorporate the combination of three wavelengths
(670nm, 728nm, and 880nm) in the treatment
groups.
The wounds were traced manually on days 4, 7,
10, and 14. These tracings were subsequently
scanned into a computer and the size of the
wounds was tracked using Sigma Scan Pro
software. Figure 1 depicts the change in wound
size over the course of the 14-day experiment.
The combination of HBO and LED (880 nm) proves
to have the greatest effect in wound healing in
terms of this qualitative assessment of wound
area. At day 7, wounds of the HBO and LED
(880nm) group are 36% smaller than those of the
control group. That size discrepancy remains
even by day 10. The LED (880nm) alone also
showed to speed wound closure. On day 7, the
LED (880 nm) treated wounds are 20% smaller than
the control wounds. By day 10, the difference
between these two groups has dropped to 12%.
This is due to the fact that there is a point
when the wounds from all of the groups will be
closed. Hence, the early differences are the
most important in terms of determining the
optimal effects of a given treatment. This can
be seen in Figure 1 at day 14 when the points
are converging due to the fact that the wounds
are healing.
Analysis of the biochemical makeup of the wounds
at days 4, 7, and 14 is currently underway. The
day 0 time point was determined by evaluating
the punch biopsy samples from the original
surgery. The levels of basic fibroblast growth
factor (FGF-2) and vascular endothelial growth
factor (VEGF) were determined using ELISA
(enzyme linked immunosorbent assay). The
changes in the VEGF concentration throughout the
14-day experiment can be seen in Figure 2. The
LED (880 nm) group experiences a VEGF peak at
day 4 much like the control group. In contrast,
the hyperoxic effect of the HBO suppresses the
day 4 peak, and instead, the HBO groups peak at
day 7. The synergistic effect of the HBO and
LED (880 nm) can be seen at day 4. The VEGF
level for the group receiving both treatments is
markedly higher at day 4 than the HBO only
group. The HBO and LED (880 nm) treated group
also experiences the day 7 peak characterized by
the HBO treatment. Hence, there is a more
uniform rise and fall to the VEGF level in the
combined treatment group as opposed to the
sudden increases seen in the control, LED only,
and HBO only groups. By day 14, the HBO treated
groups have dropped closer to the normal level
than the LED (880 nm) only or control groups.
The synergistic effects of HBO and LED (880 nm)
can be seen easily in Figure 3. The pattern of
the changes in basic fibroblast growth factor
(FGF-2) concentration is similar to that of the
VEGF data. It is clear that the LED (880 nm)
day 4 peak is higher than the day 4 peak of the
control group. These peaks can be attributed to
the hypoxic effect of the tissue ischemia
created in the surgery. The hyperoxia of the
HBO therapy has a greater effect on suppressing
the FGF-2 concentration at day 4 than the VEGF
concentration at the same time point. The
synergy of the two treatments is evident when
looking at the HBO and LED (880 nm) treated
group. The concentration of FGF-2 at day 4 is
significantly enhanced by the LED (880 nm)
treatment. Whereas, the level would normally
drop off by day 7 for a LED-only treated wound,
the HBO effect seizes control causing the
concentration of FGF-2 to plateau. Hence, an
elevated FGF-2 concentration is achieved
throughout the greater part of the 14 day
treatment with both HBO and LED (880)
therapies. Further analysis of the excised
wounds will include matrix metalloproteinase 2
and 9 (MMP-2 and MMP-9) determination by ELISA,
histological examination, and RNA extraction.
|
Figure 1. Change in wound size (%) in rat
ischemic wound model. |
LED-WOUND HEALING IN HUMAN
SUBJECTS
Preclinical and clinical
LED-Wound Healing studies were reported
previously (Whelan et al., 1999, 2000); and
additional human trials are summarized below:
Submarine atmospheres are low in oxygen and high
in carbon dioxide, which compounds the absence
of crew exposure to sunlight, making wound
healing slower than on the surface. An LED
array with 3 wavelengths combined in a single
unit (670, 720, 880 nm) was delivered to Naval
Special Warfare Group-2 in Norfolk and a data
collection system has been implemented for
musculoskeletal training injuries treated with
NASA LEDs. Data collection instruments now
include injury diagnosis, day from injury, range
of motion measured with goniometer, pain
intensity scales reported on scale 1-10,
girth-circumferential measurements in cm,
percent changes over time in all of the
aforementioned parameters, and number of
LED-treatments required for the subject to be
fit-for-full-duty (FFD). Data have also been
received from Naval Special Warfare Command
(Norfolk & San Diego) where 18-20 patients per
day are being treated with NASA-LEDs and results
indicate >40% improvement in musculoskeletal
training injuries. Data has also been received
from the USS Salt Lake City (submarine SSN 716
on Pacific deployment) reporting 50% faster (7
day) healing of lacerations in crew members
compared to untreated control healing
(approximately 14 days).
|
FIGURE 2. Change in vascular
endothelial growth factor (VEGF) concentration
(mg/mg Protein) vs. Time (Day) in rat ischemic
wound model.
FIGURE 3. Change in basic
fibroblast growth factor (FGF-2) concentration
(mg/mg Protein) vs. Time (Day) in rat ischemic
wound model. |
In addition to ischemic and
chronic wound healing, we have recently begun
using NASA LEDs to promote healing of acute oral
lesions in pediatric leukemia patients. As a
final life-saving effort, leukemia patients are
given healthy bone marrow from an HLA-matched
donor. Prior to the transplant, the patient is
given a lethal dose of chemo and radiation
therapy in order to destroy their own,
cancerous, bone marrow. Because many
chemotherapeutic drugs as well as radiation
therapy kill all rapidly dividing cells
indiscriminately, the mucosal linings of the
mouth and gastrointestinal tract are often
damaged during the treatment. As a result of
these GI effects, patients often develop ulcers
in their mouths (oral mucositis), and suffer
from nausea and diarrhea. Oral mucositis is a
significant risk for this population as it can
impair the ability to eat and drink, and poses a
risk for infection in this immunocompromized
patient. Because lasers have been shown to
speed healing of oral mucositis (Barasch, et
al., 1995), we have recently expanded the
wound-healing abilities of NASA LEDs to include
these oral lesions. Beginning on the day after
the last dose of chemotherapy, we treat one side
of the mouth with a 688nm LED at 4J/cm2 daily
until the lesions are healed. Dental clinicians
monitor the rate of healing by using an Oral
Mucositis Index (Schubert, et al., 1992) and a
Visual Analog Scale to assess mouth pain.
Although many BMT patients must receive
intravenous feeding due to their oral mucositis,
all of the patients we have treated with LEDs
have been able to eat, drink, and talk. All
have had nausea, diarrhea, and sore throats,
indicating mucositis elsewhere in their GI
tract, but their oral cavities have been
markedly less affected by mucosal ulcers. This
study has only included 10% of our target
subject number (3/30), and the data so far is
preliminary (figure1), but reports by the
attending oncologists reveal that these patients
have developed significantly less oral mucositis
than was expected, especially Patient 2 who
received Melphalan, which is notorious for
causing severe mucositis. All patients have had
Patient Controlled Analgesia (PCA) with morphine
sulfate, but all have reported that it was not
their mouths that caused them to activate
it.Further In Vitro LED Cell Growth Studies
In order to better understand the effects of
LEDs on cell growth and proliferation, we have
measured radiolabeled thymidine incorporation
in vitro in several cell lines treated with
LEDs at various wavelengths and energy levels.
As previously reported (Whelan, 2000), 3T3
fibroblasts (mouse derived skin cells) responded
extremely well to LED exposure. Cell growth
increased 150-200% over untreated controls.
Additionally, we have treated osteoblasts (rat
derived bone cells), and L6 rat skeletal muscle
cells with LEDs and have found that both
fibroblasts and particularly osteoblasts
demonstrated a growth-phase specificity to LED
treatment, responding only when cells are in the
growth phase. In these experiments, fibroblasts
and osteoblasts at a concentration of 1x104
cells/well were seeded in 24 well plates with a
well diameter of 2 square centimeters. DNA
synthesis was determined on the second, third
and fourth days in culture for both fibroblasts
(figure 1) and osteoblasts (figure 2). Exposure
to LED irradiation accelerated the growth rate
of fibroblasts and osteoblasts in culture for 2
to 3 days (growing phase), but showed no
significant change in growth rate for cells in
culture at 4 days (stationary phase). These
data are important demonstrations of cell-cell
contact inhibition, which occurs in vitro once
cell cultures approach confluence. This is
analogous in vivo to a healthy organism, which
will regenerate healing tissue, but stop further
growth when healing is complete. It is
important to demonstrate that LED treatment
accelerates this normal healing.
|
For a complete information regarding technical product
data, ordering information, and support literature, just
call our office at
1.888.874.0944, or
contact us. We
look forward to hearing from you!
|
|
 1.888.874.0944
|
|
|
|
|
$349
(+FREE
US
Shipping)
**will beat any offer
FOR
DISCOUNTS:
DISCOUNT
PRICE
CLICK HERE
(Ends
Friday)
|
 |
|
RESEARCH (
