Intro: The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, non-thermal intervention that has the potential to modulate regenerative processes is worthy of attention when searching for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a "photoceutical" for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis, and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous stem cells, as well as post-mobilization directing of stem cells will be discussed.

Background: The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, non-thermal intervention that has the potential to modulate regenerative processes is worthy of attention when searching for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a "photoceutical" for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis, and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous stem cells, as well as post-mobilization directing of stem cells will be discussed.

Abstract: Abstract The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, non-thermal intervention that has the potential to modulate regenerative processes is worthy of attention when searching for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a "photoceutical" for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis, and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous stem cells, as well as post-mobilization directing of stem cells will be discussed.

Original Source: http://www.ncbi.nlm.nih.gov/pubmed/20158898

Intro: The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, non-thermal intervention that has the potential to modulate regenerative processes is worthy of attention when searching for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a "photoceutical" for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis, and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous stem cells, as well as post-mobilization directing of stem cells will be discussed.

Background: The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, non-thermal intervention that has the potential to modulate regenerative processes is worthy of attention when searching for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a "photoceutical" for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis, and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous stem cells, as well as post-mobilization directing of stem cells will be discussed.

Abstract: Abstract The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, non-thermal intervention that has the potential to modulate regenerative processes is worthy of attention when searching for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a "photoceutical" for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis, and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous stem cells, as well as post-mobilization directing of stem cells will be discussed.

Original Source: http://www.ncbi.nlm.nih.gov/pubmed/20158898

 Abstract

The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue

healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, nonthermal

intervention that has the potential to modulate regenerative processes is worthy of attention when searching

for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a

“photoceutical” for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis,

and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous

stem cells, as well as post-mobilization directing of stem cells will be discussed.

Introduction (Personal Perspective)

We came upon the field of low level laser (LLL) therapy

by accident. One of our advisors read a press release

about a company using this novel technology of specific

light wavelengths to treat stroke. Given the possible role

of stem cells in post-stroke regeneration, we decided to

cautiously investigate. As a background, it should be

said that our scientific team has been focusing on the

area of cord blood banking and manufacturing of disposables

for processing of adipose stem cells for the past 3

years. Our board has been interested in strategically

refocusing the company from services-oriented into a

more research-focused model. An unbiased exploration

into the various degenerative conditions that may be

addressed by our existing know-how led us to explore

the condition of chronic obstructive pulmonary disease

(COPD), an umbrella term covering chronic bronchitis

and emphysema, which is the 4th largest cause of death

in the United States. As a means of increasing our probability

of success in treatment of this condition, the

decision was made to develop an adjuvant therapy that

would augment stem cell activity. The field of LLL therapy

attracted us because it appeared to be relatively

unexplored scientific territory for which large amounts

of clinical experience exist. Unfortunately, it was difficult

to obtain the cohesive “state-of-the-art” description of

the molecular/cellular mechanisms of this therapy in

reviews that we have searched. Therefore we sought in

this mini-review to discuss what we believe to be relevant

to investigators attracted by the concept of “regenerative

photoceuticals”. Before presenting our synthesis

of the field, we will begin by describing our rationale for

approaching COPD with the autologous stem cell based

approaches we are developing.

COPD as an Indication for Stem Cell Therapy

COPD possesses several features making it ideal for

stem cell based interventions: a) the quality of life and

lack of progress demands the ethical exploration of

novel approaches. For example, bone marrow stem cells

have been used in over a thousand cardiac patients with

some indication of efficacy [1,2]. Adipose-based stem

cell therapies have been successfully used in thousands

of race-horses and companion animals without adverse

effects [3], as well as numerous clinical trials are

ongoing and published human data reports no adverse

effects (reviewed in ref [4]). Unfortunately, evaluation of

stem cell therapy in COPD has lagged behind other

areas of regenerative investigation; b) the underlying

cause of COPD appears to be inflammatory and/or

immunologically mediated. The destruction of alveolar

tissue is associated with T cell reactivity [5,6], pathological

pulmonary macrophage activation [7], and auto-antibody

production [8]. Mesenchymal stem cells have been

demonstrated to potently suppress autoreactive T cells

[9,10], inhibit macrophage activation [11], and autoantibody

responses [12]. Additionally, mesenchymal stem

cells can be purified in high concentrations from adipose

stromal vascular tissue together with high concentrations of T regulatory cells [4], which in animal

models are approximately 100 more potent than peripheral

T cells at secreting cytokines therapeutic for COPD

such as IL-10 [13,14]. Additionally, use of adipose

derived cells has yielded promising clinical results in

autoimmune conditions such as multiple sclerosis [4];

and c) Pulmonary stem cells capable of regenerating

damaged parenchymal tissue have been reported [15].

Administration of mesenchymal stem cells into neonatal

oxygen-damaged lungs, which results in COPD-like

alveoli dysplasia, has been demonstrated to yield

improvements in two recent publications [16,17].

Based on the above rationale for stem cell-based

COPD treatments, we began our exploration into this

area by performing several preliminary experiments and

filing patents covering combination uses of stem cells

with various pharmacologically available antiinflammatories,

as well as methods of immune modulation. These

have served as the basis for two of our pipeline candidates,

ENT-111, and ENT-894. As a commerciallyoriented

organization, we needed to develop a therapeutic

candidate that not only has a great potential for efficacy,

but also can be easily implemented as part of the

standard of care. Our search led us to the area of low

level laser (LLL) therapy. From our initial perception as

neophytes to this field, the area of LLL therapy has been

somewhat of a medical mystery. A pubmed search for

“low level laser therapy” yields more than 1700 results,

yet before stumbling across this concept, none of us, or

our advisors, have ever heard of this area of medicine.

On face value, this field appeared to be somewhat of a

panacea: clinical trials claiming efficacy for conditions

ranging from alcoholism [18], to sinusitis [19], to

ischemic heart disease [20]. Further confusing was that

many of the studies used different types of LLL-generating

devices, with different parameters, in different model

systems, making comparison of data almost impossible.

Despite this initial impression, the possibility that a simple,

non-invasive methodology could exist that augments

regenerative potential in a tissue-focused manner

became very enticing to us. Specific uses envisioned, for

which intellectual property was filed included using light

to concentrate stem cells to an area of need, to modulate

effects of stem cells once they are in that specific

area, or even to use light together with other agents to

modulate endogenous stem cells.

The purpose of the current manuscript is to overview

some of the previous work performed in this area that was

of great interest to our ongoing work in regenerative medicine.

We believe that greater integration of the area of

LLL with current advancements in molecular and cellular

biology will accelerate medical progress. Unfortunately, in

our impression to date, this has been a very slow process.

What is Low Level Laser Irradiation?

Lasers (Light amplification by stimulated emission of

radiation) are devices that typically generate electromagnetic

radiation which is relatively uniform in wavelength,

phase, and polarization, originally described by Theodore

Maiman in 1960 in the form of a ruby laser [21]. These

properties have allowed for numerous medical applications

including uses in surgery, activation of photodynamic

agents, and various ablative therapies in cosmetics that are

based on heat/tissue destruction generated by the laser

beam [22-24]. These applications of lasers are considered

“high energy” because of their intensity, which ranges

from about 10-100 Watts. The subject of the current

paper will be another type of laser approach called low

level lasers (LLL) that elicits effects through non-thermal

means. This area of investigation started with the work of

Mester et al who in 1967 reported non-thermal effects of

lasers on mouse hair growth [25]. In a subsequent study

[26], the same group reported acceleration of wound healing

and improvement in regenerative ability of muscle

fibers post wounding using a 1 J/cm2 ruby laser. Since

those early days, numerous in vitro and in vivo studies

have been reported demonstrating a wide variety of therapeutic

effects involving LLL, a selected sample of which

will be discussed below. In order to narrow our focus of

discussion, it is important to first begin by establishing the

current definition of LLL therapy. According to Posten et

al [27], there are several parameters of importance: a)

Power output of laser being 10-3 to 10-1 Watts; b) Wavelength

in the range of 300-10,600 nm; c) Pulse rate from 0,

meaning continuous to 5000 Hertz (cycles per second); d)

intensity of 10-2-10 W/cm(2) and dose of 0.01 to 100 J/

cm2. Most common methods of administering LLL radiation

include lasers such as ruby (694 nm), Ar (488 and 514

nm), He-Ne (632.8 nm), Krypton (521, 530, 568, and 647

nm), Ga-Al-As (805 or 650 nm), and Ga-As (904 nm).

Perhaps one of the most distinguishing features of LLL

therapy as compared to other photoceutical modalities is

that effects are mediated not through induction of thermal

effects but rather through a process that is still not clearly

defined called “photobiostimulation”. It appears that this

effect of LLL is not depend on coherence, and therefore

allows for use of non-laser light generating devices such as

inexpensive Light Emitting Diode (LED) technology [28].

To date several mechanisms of biological action have

been proposed, although none are clearly established.

These include augmentation of cellular ATP levels [29],

manipulation of inducible nitric oxide synthase (iNOS)

activity [30,31], suppression of inflammatory cytokines

such as TNF-alpha, IL-1beta, IL-6 and IL-8 [32-36],

upregulation of growth factor production such as PDGF,

IGF-1, NGF and FGF-2 [36-39], alteration of mitochondrial

membrane potential [29,40-42] due to chromophores found in the mitochondrial respiratory

chain [43,44] as reviewed in [45], stimulation of protein

kinase C (PKC) activation [46], manipulation of NF-!B

activation [47], direct bacteriotoxic effect mediated by

induction of reactive oxygen species (ROS) [48], modification

of extracellular matrix components [49], inhibition

of apoptosis [29], stimulation of mast cell

degranulation [50], and upregulation of heat shock proteins

[51]. Unfortunately these effects have been demonstrated

using a variety of LLL devices in noncomparable

models. To add to confusion, dose-dependency

seems to be confined to such a narrow range or

does not seem to exist in that numerous systems therapeutic

effects disappear with increased dose.

In vitro studies of LLL

In areas of potential phenomenology, it is important to

begin by assessing in vitro studies reported in the literature

in which reproducibility can be attained with some

degree of confidence, and mechanistic dissection is simpler

as compared with in vivo systems. In 1983, one of

the first studies to demonstrate in vitro effects of LLL

was published. The investigators used a helium neon

(He-Ne) laser to generate a visible red light at 632.8 nm

for treatment of porcine granulosa cells. The paper

described upregulation of metabolic and hormone-producing

activity of the cells when exposed for 60 seconds

to pulsating low power (2.8 mW) irradiation [52]. The

possibility of modulating biologically-relevant signaling

proteins by LLL was further assessed in a study using an

energy dose of 1.5 J/cm2 in cultured keratinocytes.

Administration of He-Ne laser emitted light resulted in

upregulated gene expression of IL-1 and IL-8 [53]. Production

of various growth factors in vitro suggests the

possibility of enhanced cellular mitogenesis and mobility

as a result of LLL treatment. Using a diode-based

method to generate a similar wavelength to the He-Ne

laser (363 nm), Mvula et al reported in two papers that

irradiation at 5 J/cm2 of adipose derived mesenchymal

stem cells resulted in enhanced proliferation, viability

and expression of the adhesion molecule beta-1 integrin

as compared to control [54,55]. In agreement with possible

regenerative activity based on activation of stem

cells, other studies have used an in vitro injury model to

examine possible therapeutic effects. Migration of fibroblasts

was demonstrated to be enhanced in a “wound

assay” in which cell monolayers are scraped with a pipette

tip and amount of time needed to restore the

monolayer is used as an indicator of “healing”. The cells

exposed to 5 J/cm2 generated by an He-Ne laser

migrated rapidly across the wound margin indicating a

stimulatory or positive influence of phototherapy.

Higher doses (10 and 16 J/cm2) caused a decrease in

cell viability and proliferation with a significant amount

of damage to the cell membrane and DNA [56]. In

order to examine whether LLL may positively affect

healing under non-optimal conditions that mimic clinical

situations treatment of fibroblasts from diabetic animals

was performed. It was demonstrated that with the

He-Ne laser dosage of 5 J/cm2 fibroblasts exhibited an

enhanced migration activity, however at 16 J/cm2 activity

was negated and cellular damage observed [57]. Thus

from these studies it appears that energy doses from 1.5

J/cm2 to 5 J/cm2 are capable of eliciting “biostimulatory

effects” in vitro in the He-Ne-based laser for adherent

cells that may be useful in regeneration such as fibroblasts

and mesenchymal stem cells.

Studies have also been performed in vitro on immunological

cells. High intensity He-Ne irradiation at 28

and 112 J/cm2 of human peripheral blood mononuclear

cells, a heterogeneous population of T cells, B cells, NK

cells, and monocytes has been described to induce chromatin

relaxation and to augment proliferative response

to the T cell mitogen phytohemaglutin [58]. In human

peripheral blood mononuclear cells (PBMC), another

group reported in two papers that interleukin-1 alpha

(IL-1 alpha), tumor necrosis factor-alpha (TNF-alpha),

interleukin-2 (IL-2), and interferon-gamma (IFNgamma)

at a protein and gene level in PBMC was

increased after He-Ne irradiation at 18.9 J/cm2 and

decreased with 37.8 J/cm2 [59,60]. Stimulation of human

PBMC proliferation and murine splenic lymphocytes

was also reported with He-Ne LLL [61,62]. In terms of

innate immune cells, enhanced phagocytic activity of

murine macrophages have been reported with energy

densities ranging from 100 to 600 J/cm2, with an optimal

dose of 200 J/cm2 [63]. Furthermore, LLL has been

demonstrated to augment human monocyte killing

mycobacterial cells at similar densities, providing a functional

correlation [64].

Thus from the selected in vitro studies discussed, it

appears that modulation of proliferation and soluble factor

production by LLL can be reliably reproduced. However

the data may be to some extent contradictory. For

example, the over-arching clinical rationale for use of

LLL in conditions such as sinusitis [65], arthritis [66,67],

or wound healing [68] is that treatment is associated

with anti-inflammatory effects. However the in vitro studies

described above suggested LLL stimulates proinflammatory

agents such as TNF-alpha or IL-1 [59,60].

This suggests the in vivo effects of LLL may be very

complex, which to some extent should not be surprising.

Factors affecting LLL in vivo actions would include

degree of energy penetration through the tissue, the various

absorption ability of cells in the various tissues, and

complex chemical changes that maybe occurring in

paracrine/autocrine manner. Perhaps an analogy to the

possible discrepancy between LLL effects in vitro versus in vivo may be made with the medical practice of extracorporeal

ozonation of blood. This practice is similar to

LLL therapy given that it is used in treatment of conditions

such as atherosclerosis, non-healing ulcers, and

various degenerative conditions, despite no clear

mechanistic understanding [69-71]. In vitro studies have

demonstrated that ozone is a potent oxidant and inducer

of cell apoptosis and inflammatory signaling [72-74].

In contrast, in vivo systemic changes subsequent to

administration of ozone or ozonized blood in animal

models and patients are quite the opposite. Numerous

investigators have published enhanced anti-oxidant

enzyme activity such as elevations in Mg-SOD and glutathione-

peroxidase levels, as well as diminishment of

inflammation-associated pathology [75-78]. Regardless

of the complexity of in vivo situations, the fact that

reproducible, in vitro experiments, demonstrate a biological

effect provided support for us that there is some

basis for LLL and it is not strictly an area of

phenomenology.

Animal Studies with LLL

As early as 1983, Surinchak et al reported in a rat skin

incision healing model that wounds exposed He-Ne

radiation of fluency 2.2 J/cm2 for 3 min twice daily for

14 days demonstrated a 55% increase in breaking

strength over control rats. Interestingly, higher doses

yielded poorer healing [79]. This application of laser

light was performed directly on shaved skin. In a contradictory

experiment, it was reported that rats irradiated

for 12 days with four levels of laser light (0.0, 0.47, 0.93,

and 1.73 J/cm2) a possible strengthening of wounds tension

was observed at the highest levels of irradiation

(1.73 J/cm2), however it did not reach significance when

analyzed by resampling statistics [80]. In another

wound-healing study Ghamsari et al reported accelerated

healing in the cranial surface of teats in dairy cows

by administration of He-Ne irradiation at 3.64 J/cm2

dose of low-level laser, using a helium-neon system with

an output of 8.5 mW, continuous wave [81]. Collagen

fibers in LLL groups were denser, thicker, better

arranged and more continuous with existing collagen

fibers than those in non-LLL groups. The mean tensile

strength was significantly greater in LLL groups than in

non-LLL groups [82]. In the random skin flap model,

the use of He-Ne laser irradiation with 3 J/cm2 energy

density immediately after the surgery and for the four

subsequent days was evaluated in 4 experimental

groups: Group 1 (control) sham irradiation with He-Ne

laser; Group 2 irradiation by punctual contact technique

on the skin flap surface; Group 3 laser irradiation surrounding

the skin flap; and Group 4 laser irradiation

both on the skin flap surface and around it. The percentage

of necrotic area of the four groups was determined

on day 7-post injury. The control group had an average

necrotic area of 48.86%; the group irradiated on the skin

flap surface alone had 38.67%; the group irradiated

around the skin flap had 35.34%; and the group irradiated

one the skin flap surface and around it had

22.61%. All experimental groups reached statistically significant

values when compared to control [83]. Quite

striking results were obtained in an alloxan-induced diabetes

wound healing model in which a circular 4 cm2

excisional wound was created on the dorsum of the diabetic

rats. Treatment with He-Ne irradiation at 4.8 J/

cm2 was performed 5 days a week until the wound

healed completely and compared to sham irradiated animals.

The laser-treated group healed on average by the

18th day whereas, the control group healed on average

by the 59th day [84].

In addition to mechanically-induced wounds, beneficial

effects of LLL have been obtained in burn-wounds

in which deep second-degree burn wounds were

induced in rats and the effects of daily He-Ne irradiation

at 1.2 and 2.4 J/cm2 were assessed in comparison to

0.2% nitrofurazone cream. The number of macrophages

at day 16, and the depth of new epidermis at day 30,

was significantly less in the laser treated groups in comparison

with control and nitrofurazone treated groups.

Additionally, infections with S. epidermidis and S. aureus

were significantly reduced [85].

While numerous studies have examined dermatological

applications of LLL, which may conceptually be

easier to perform due to ability to topically apply light,

extensive investigation has also been made in the area

of orthopedic applications. Healing acceleration has

been observed in regeneration of the rat mid-cortical

diaphysis of the tibiae, which is a model of post-injury

bone healing. A small hole was surgically made with a

dentistry burr in the tibia and the injured area and LLL

was administered over a 7 or 14 day course transcutaneously

starting 24 h from surgery. Incident energy density

dosages of 31.5 and 94.5 J/cm2 were applied during

the period of the tibia wound healing. Increased angiogenesis

was observed after 7 days irradiation at an

energy density of 94.5 J/cm2, but significantly decreased

the number of vessels in the 14-day irradiated tibiae,

independent of the dosage [86]. In an osteoarthritis

model treatment with He-Ne resulted in augmentation

of heat shock proteins and pathohistological improvement

of arthritic cartilage [87]. The possibility that a

type of preconditioning response is occurring, which

would involve induction of genes such as hemoxygenase-

1 [88], remains to be investigated. Effects of LLL

therapy on articular cartilage were confirmed by another

group. The experiment consisted of 42 young Wistar

rats whose hind limbs were operated on in order to

immobilize the knee joint. One week after operation they were assigned to three groups; irradiance 3.9 W/

cm2, 5.8 W/cm2, and sham treatment. After 6 times of

treatment for another 2 weeks significantpreservation of

articular cartilage stiffness with 3.9 and 5.8 W/cm2 therapy

was observed [89].

Muscle regeneration by LLL was demonstrated in a rat

model of disuse atrophy in which eight-week-old rats

were subjected to hindlimb suspension for 2 weeks,

after which they were released and recovered. During

the recovery period, rats underwent daily LLL irradiation

(Ga-Al-As laser; 830 nm; 60 mW; total, 180 s) to

the right gastrocnemius muscle through the skin. After

2-weeks the number of capillaries and fibroblast growth

factor levels exhibited significant elevation relative to

those of the LLL-untreated muscles. LLL treatment

induced proliferation in satellite cells as detected by

BRdU [90].

Other animal studies of LLL have demonstrated

effects in areas that appear unrelated such as suppression

of snake venom induced muscle death [91],

decreasing histamine-induced vasospasms [92], inhibition

of post-injury restenosis [93], and immune stimulation

by thymic irradiation [94].

Clinical Studies Using LLL

Growth factor secretion by LLL and its apparent regenerative

activities have stimulated studies in radiationinduced

mucositis. A 30 patient randomized trial of carcinoma

patients treated by radiotherapy alone (65 Gy at

a rate of 2 Gy/fraction, 5 fractions per week) without

prior surgery or concomitant chemotherapy suffering

from radiation-induced mucositis was performed using a

He-Ne 60 mW laser. Grade 3 mucositis occured with a

frequency of 35.2% in controls and at 7.6% of treated

patients. Furthermore, a decrease in “severe pain” (grade

3) was observed in that 23.8% in the control group

experienced this level of pain, as compared to 1.9% in

the treatment group [95]. A subsequent study reported

similar effects [96].

Healing ability of lasers was also observed in a study

of patients with gingival flap incisions. Fifty-eight extraction

patients had one of two gingival flap incisions lased

with a 1.4 mW He-Ne (670 nm) at 0.34 J/cm2. Healing

rates were evaluated clinically and photographically.

Sixty-nine percent of the irradiated incisions healed faster

than the control incisions. No significant difference

in healing was noted when patients were compared by

age, gender, race, and anatomic location of the incision

[97]. Another study evaluating healing effects of LLL in

dental practice examined 48 patients subjected to surgical

removal of their lower third molars. Treated patients

were administered Ga-Al-As diode generated 808 nm at

a dose of 12 J. The study demonstrated that extraoral

LLL is more effective than intraoral LLL, which was

more effective than control for the reduction of postoperative

trismus and swelling after extraction of the

lower third molar [98].

Given the predominance of data supporting fibroblast

proliferative ability and animal wound healing effects of

LLL therapy, a clinical trial was performed on healing of

ulcers. In a double-blinded fashion 23 diabetic leg ulcers

from 14 patients were divided into two groups. Phototherapy

was applied (<1.0 J/cm2) twice per week, using a

Dynatron Solaris 705(R) LED device that concurrently

emits 660 and 890 nm energies. At days 15, 30, 45, 60,

75, and 90 mean ulcer granulation and healing rates

were significantly higher for the treatment group as

compared to control. By day 90, 58.3% of the ulcers in

the LLL treated group were fully healed and 75%

achieved 90-100% healing. In the placebo group only

one ulcer healed fully [68].

As previously mentioned, LLL appears to have some

angiogenic activity. One of the major problems in coronary

artery disease is lack of collateralization. In a 39

patient study advanced CAD, two sessions of irradiation

of low-energy laser light on skin in the chest area from

helium-neon B1 lasers. The time of irradiation was 15

minutes while operations were performed 6 days a week

for one month. Reduction in Canadian Cardiology

Society (CCS) score, increased exercise capacity and

time, less frequent angina symptoms during the treadmill

test, longer distance of 6-minute walk test and a

trend towards less frequent 1 mm ST depression lasting

1 min during Holter recordings was noted after therapy

[99].

Perhaps one of the largest clinical trials with LLL was

the NEST trial performed by Photothera. In this double

blind trial 660 stroke patients were recruited and randomized:

331 received LLL and 327 received sham. No

prespecified test achieved significance, but a post hoc

analysis of patients with a baseline National Institutes of

Health Stroke Scale score of <16 showed a favorable

outcome at 90 days on the primary end point (P <

0.044) [100]. Currently Photothera is in the process of

repeating this trial with modified parameters.

Relevance of LLL to COPD

A therapeutic intervention in COPD would require

addressing the issues of inflammation and regeneration.

Although approaches such as administration of bone marrow

stem cells, or fat derived cellular components have

both regenerative and anti-inflammatory activity in animal

models, the need to enhance their potency for clinical

applications can be seen in the recent Osiris’s COPD trial

interim data which reported no significant improvement

in pulmonary function [101]. Accordingly, we sought to

develop a possible rationale for how LLL may be useful as

an adjunct to autologous stem cell therapy.

Table 1 Examples of LLL Properties Relevant to COPD

COPD

Property

LLL Experiment LLL Details Ref

Inflammation In vivo. Decreased joint inflammation in zymosan-induced

arthritis

Semiconductor laser (685 nm and 830 nm) at (2.5 J/cm2)

In vitro. Suppression of LPS-induced bronchial inflammation and

TNF-alpha.

655 nm at of 2.6 J/cm2

In vivo. Carrageenan-induced pleurisy had decreased leukocyte

infiltration and cytokine (TNF-alpha, IL-6, and MCP)

660 nm at 2.1 J/cm2

In vitro. LPS stimulated Raw 264.7 monocytes had reduced gene

expression of MCP-1, IL-1 and IL-6

780 nm diode laser at 2.2 J/cm2)

In vivo. Suppression of LPS-stimulated neutrophil influx,

myeloperoxidase activity and IL-1beta in bronchoalveolar lavage

fluid.

660 nm diode laser at 7.5 J/cm2

In vitro. Inhibition of TNF-alpha induced IL-1, IL-8 and TNF-alpha

mRNA in human synoviocytes

810 nm (5 J/cm2) suppressed IL-1 and TNF, (25 J/cm2) also

suppressed IL-8

In vivo. Reduction of TNF-alpha in diaphragm muscle after

intravenous LPS injection.

4 sessions in 24 h with diode Ga-AsI-Al laser of 650 nm and

a total dose of 5.2 J/cm2

In vivo. Inhibition of LPS induced peritonitis and neutrophil influx 3 J/cm2 and 7.5 J/cm2

Growth Factor Production

In vivo. Upregulation of TGF-b and PDGF in rat gingiva after

incision.

He-Ne laser (632.8 nm) at a dose of 7.5 J/cm2

In vitro. Osteoblast-like cells were isolated from fetal rat calvariae

had increased IGF-1

Ga-Al-As laser (830 nm) at (3.75 J/cm2).

In vitro. Upregulated production of IGF-1 and FGF-2 in human

gingival fibroblasts.

685 nm, for 140 s, 2 J/cm2

Angiogenesis

In vivo. Increased fiber to capillary ratio in rabbits with ligated

femoral arteries.

Gallium-aluminum-arsenide (Ga-Al-As) diode laser, 904 nm

and power of 10 mW

In vitro. Stimulation of HUVEC proliferation by conditioned media

from LLL-treated T cells

820 nm at 1.2 and 3.6 J/cm2.

In vitro. 7-fold increased production of VEGF by cardiomyocytes,

1.6-fold increase by smooth muscle cells (SMC) and fibroblasts.

Supernatant of SMC had increased HUVEC-stimulating potential.

He:Ne continuous wave laser (632 nm). 0.5 J/cm2 for SMC,

2.1 J/cm2 for fibroblasts and 1.05 J/cm2 for cardiomyocytes.

In vitro. Direct stimulation of HUVEC proliferation 670 nm diode device at 2 and 8 J/cm2

Direct Stem Cell Effects

In vivo. LLL precondition significantly enhanced early cell survival

rate by 2-fold, decreased the apoptotic percentage of implanted

BMSCs in infarcted myocardium and increased the number of

newly formed capillaries.

635 nm at 0.96 J/cm2

In vitro. LLL stimulated MSC proliferation, VEGF and NGF

production, and myogenic differentiation after 5-aza induction.

635 nm diode laser at 0.5 J/cm2 for MSC proliferation, 5 J/

cm2 for VEGF and NGF production and for augmentation of

induced myogenic differentiation

In vitro. Increased proliferation of rat MSC. Red light LED 630 nm at 2 and 4 J/cm(2)

In vitro. Augmented proliferation of bone marrow and cardiac

specific stem cells.

GA-Al-As 810 nm at 1 and 3 J/cm2

In vitro/In vivo. Administration of LLL-treated MSC resulted 53%

reduction in infarct size, 5- and 6.3-fold significant increase in cell

density that positively immunoreacted to BrdU and c-kit,

respectively, and 1.4- and 2-fold higher level of angiogenesis and

vascular endothelial growth factor, respectively, when compared

to non-laser-treated implanted cells

Ga-Al-As laser (810 nm wavelength), 1 J/cm2

In vitro. Enhanced proliferation of adipose derived MSC in

presence of EGF.

636 nm diode, 5 J/cm2

Lin et al. Journal of Translational Medicine 2010, 8:16

http://www.translational-medicine.com/content/8/1/16

Table 1: Examples of LLL Properties Relevant to COPD (Continued)

In vitro. Enhanced proliferation and beta-1 integrin expression of

adipose derived MSC.

635 nm diode laser, at 5 J/cm2

Clinical. 660 stroke patients: 331 received LLL and 327 received

sham. No prespecified test achieved significance, but a post hoc

analysis of patients with a baseline National Institutes of Health

Stroke Scale score of <16 showed a favorable outcome at 90

days on the primary end point (P < 0.044).

808 nm. No density disclosed.

 

Table 1 depicts some of the properties of LLL that provide

a rationale for the combined use with stem cells. One

of the basic properties of LLL seems to be ability to inhibit

inflammation at the level of innate immune activation.

Representative studies showed that LLL was capable of

suppressing inflammatory genes and/or pathology after

administration of lipopolysaccharide (LPS) as a stimulator

of monocytes [102] and bronchial cells [34], in vitro, and

leukocyte infiltration in vivo [103,104]. Inflammation

induced by other stimulators such as zymosan, carrageenan,

and TNF-alpha was also inhibited by LLL

[32,105,106]. Growth factor stimulating activity of LLL

was demonstrated in both in vitro and in vivo experiments

in which augmentation of FGF-2, PDGF and IGF-1 was

observed [36,37,107]. Endogenous production of these

growth factors may be useful in regeneration based on

activation of endogenous pulmonary stem cells [108,109].

Another aspect of LLL activities of relevance is ability to

stimulate angiogenesis. In COPD, the constriction of

blood vessels as a result of poor oxygen uptake is results

in a feedback loop culminating in pulmonary hypertension.

Administration of angiogenic factors has been

demonstrated to be beneficial in several animal models of

pulmonary pathology [110,111]. The ability of LLL to

directly induce proliferation of HUVEC cells [112], as well

as to augment production of angiogenic factors such as

VEGF [113], supports the possibility of creation of an

environment hospitable to neoangiogenesis which is optimal

for stem cell growth. In fact, a study demonstrated in

vivo induction of neocapillary formation subsequent to

LLL administration in a hindlimb ischemia model [114].

The critical importance of angiogenesis in stem cell

mediated regeneration has previously been demonstrated

in the stroke model, where the major therapeutic activity

of exogenous stem cells has been attributed to angiogenic

as opposed to transdifferentiation effects [115].

Direct evidence of LLL stimulating stem cells has been

obtained using mesenchymal stem cells derived both

from the bone marrow and from the adipose tissue

[116,117]. Interestingly in vivo administration of LLL stimulated

MSC has resulted in 50% decrease in cardiac

infarct size [118]. Clinical translation of LLL has been

performed in the area of stroke, in which a 660 patient

trial demonstrated statistically significant effects in post

trial subset analysis [100].

Conclusions

Despite clinical use of LLL for decades, the field is still

in its infancy. As is obvious from the wide variety of

LLL sources, frequencies, and intensities used, no standard

protocols exist. The ability of LLL to induce

growth factor production, inhibition of inflammation,

stimulation of angiogenesis, and direct effects on stem

cells suggests the urgent need for combining this modality

with regenerative medicine, giving birth to the new

field of “regenerative photoceuticals”. Development of a

regenerative treatment for COPD as well as for other

degenerative diseases would be of considerable benefit.

Regarding COPD, such treatment would be life-saving/

life extending for thousands of affected individuals.

Ceasing smoking or not starting to smoke would considerably

impact this disease.

Acknowledgements

The authors thank Victoria Dardov and Matthew Gandjian for critical

discussions and input.

Author details

1Entest BioMedical, San Diego, CA, USA. 2Georgetown Dermatology,

Washington DC, USA. 3Cromos Pharma Services, Longview, WA, USA. 4Center

for the Study of Natural Oncology, Del Mar, CA, USA. 5Department of

Hematology and Medical Oncology, St Francis Hospital and Medical Center,

Hartford, CT, USA. 6Moores Cancer Center, University of California San Diego,

CA, USA. 7Department of Cardiothoracic Surgery, University of Utah, Salt

Lake City, UT, USA.

Authors’ contributions

FL, SFJ, DTA, FR, VB, VG, CAD, RDNC, ANP, EC, DRK contributed to literature

review, analysis and discussion, synthesis of concepts, writing of the

manuscript and proof-reading of the final draft.

Competing interests

David R Koos is a shareholder, as well as Chairman and CEO of Entest Bio.

Feng Lin is research director of Entest Bio. All other authors declare no

competing interest.

Received: 7 January 2010

Accepted: 16 February 2010 Published: 16 February 2010

References

1. Abdel-Latif A, Bolli R, Tleyjeh IM, Montori VM, Perin EC, Hornung CA, Zuba-

Surma EK, Al-Mallah M, Dawn B: Adult bone marrow-derived cells for

cardiac repair: a systematic review and meta-analysis. Arch Intern Med

2007, 167:989-997.

2. Martin-Rendon E, Brunskill SJ, Hyde CJ, Stanworth SJ, Mathur A, Watt SM:

Autologous bone marrow stem cells to treat acute myocardial infarction:

a systematic review. Eur Heart J 2008, 29:1807-1818.

3. Vet-Stem Regenerative Veterinary Medicine. http://www.vet-stem.com.

4. Riordan NH, Ichim TE, Min WP, Wang H, Solano F, Lara F, Alfaro M,

Rodriguez JP, Harman RJ, Patel AN, Murphy MP, Lee RR, Minev B: Nonexpanded

adipose stromal vascular fraction cell therapy for multiple

sclerosis. J Transl Med 2009, 7:29.

5. Motz GT, Eppert BL, Sun G, Wesselkamper SC, Linke MJ, Deka R,

Borchers MT: Persistence of lung CD8 T cell oligoclonal expansions upon

smoking cessation in a mouse model of cigarette smoke-induced

emphysema. J Immunol 2008, 181:8036-8043.

6. Maeno T, Houghton AM, Quintero PA, Grumelli S, Owen CA, Shapiro SD:

CD8+ T Cells are required for inflammation and destruction in cigarette

smoke-induced emphysema in mice. J Immunol 2007, 178:8090-8096.

7. Woodruff PG, Koth LL, Yang YH, Rodriguez MW, Favoreto S, Dolganov GM,

Paquet AC, Erle DJ: A distinctive alveolar macrophage activation state

induced by cigarette smoking. Am J Respir Crit Care Med 2005,

172:1383-1392.

8. Stefanska AM, Walsh PT: Chronic obstructive pulmonary disease: evidence

for an autoimmune component. Cell Mol Immunol 2009, 6:81-86.

9. Gonzalez-Rey E, Gonzalez MA, Varela N, O’Valle F, Hernandez-Cortes P,

Rico L, Buscher D, Delgado M: Human adipose-derived mesenchymal

stem cells reduce inflammatory and T cell responses and induce

regulatory T cells in vitro in rheumatoid arthritis. Ann Rheum Dis

69:241-248.

10. Lepelletier Y, Lecourt S, Arnulf B, Vanneaux V, Fermand JP, Menasche P,

Domet T, Marolleau JP, Hermine O, Larghero J: Galectin-1 and

Semaphorin-3A are two soluble factors conferring T cell

immunosuppression to bone marrow mesenchymal stem cell. Stem Cells

Dev 2009.

11. Tsyb AF, Petrov VN, Konoplyannikov AG, Saypina EV, Lepechina LA,

Kalsina S, Semenkova IV, Agaeva EV: In vitro inhibitory effect of

mesenchymal stem cells on zymosan-induced production of reactive

oxygen species. Bull Exp Biol Med 2008, 146:158-164.

12. Sun L, Akiyama K, Zhang H, Yamaza T, Hou Y, Zhao S, Xu T, Le A, Shi S:

Mesenchymal stem cell transplantation reverses multiorgan dysfunction

in systemic lupus erythematosus mice and humans. Stem Cells 2009,

27:1421-1432.

13. Feuerer M, Herrero L, Cipolletta D, Naaz A, Wong J, Nayer A, Lee J,

Goldfine AB, Benoist C, Shoelson S, Mathis D: Lean, but not obese, fat is

enriched for a unique population of regulatory T cells that affect

metabolic parameters. Nat Med 2009, 15:930-939.

14. Ogawa Y, Duru EA, Ameredes BT: Role of IL-10 in the resolution of airway

inflammation. Curr Mol Med 2008, 8:437-445.

15. Serrano-Mollar A, Nacher M, Gay-Jordi G, Closa D, Xaubet A, Bulbena O:

Intratracheal transplantation of alveolar type II cells reverses bleomycininduced

lung fibrosis. Am J Respir Crit Care Med 2007, 176:1261-1268.

16. Aslam M, Baveja R, Liang OD, Fernandez-Gonzalez A, Lee C, Mitsialis SA,

Kourembanas S: Bone marrow stromal cells attenuate lung injury in a

murine model of neonatal chronic lung disease. Am J Respir Crit Care Med

2009, 180:1122-1130.

17. van Haaften T, Byrne R, Bonnet S, Rochefort GY, Akabutu J, Bouchentouf M,

Rey-Parra GJ, Galipeau J, Haromy A, Eaton F, Chen M, Hashimoto K,

Abley D, Korbutt G, Archer SL, Thébaud B: Airway delivery of

mesenchymal stem cells prevents arrested alveolar growth in neonatal

lung injury in rats. Am J Respir Crit Care Med 2009, 180:1131-1142.

18. Zalewska-Kaszubska J, Obzejta D: Use of low-energy laser as adjunct

treatment of alcohol addiction. Lasers Med Sci 2004, 19:100-104.

19. Nikitin AV, Esaulenko IE, Shatalova OL: [Effectiveness of laser puncture in

elderly patients with bronchial asthma accompanied by chronic

rhinosinusitis]. Adv Gerontol 2008, 21:424-426.

20. Vasil’ev AP, Strel’tsova NN, Iu Senatorov N: [Laser irradiation in the

treatment of ischemic heart disease]. Vopr Kurortol Fizioter Lech Fiz Kult

2001, 10-13.

21. Maiman TH: Stimulated optical radiation in Ruby. Nature 187:493.

22. Roy D: Ablative facial resurfacing. Dermatol Clin 2005, 23:549-559, viii.

23. Brown MC: An evidence-based approach to patient selection for laser

vision correction. J Refract Surg 2009, 25:S661-667.

24. Brancaleon L, Moseley H: Laser and non-laser light sources for

photodynamic therapy. Lasers Med Sci 2002, 17:173-186.

25. Mester ESB, Tota JG: “Effect of laser on hair growth of mice”. Kiserl

Orvostud 1967, 19:628-631.

26. Mester E, Korenyi-Both A, Spiry T, Tisza S: The effect of laser irradiation on

the regeneration of muscle fibers (preliminary report). Z Exp Chir 1975,

8:258-262.

27. Posten W, Wrone DA, Dover JS, Arndt KA, Silapunt S, Alam M: Low-level

laser therapy for wound healing: mechanism and efficacy. Dermatol Surg

2005, 31:334-340.

28. Vladimirov YA, Osipov AN, Klebanov GI: Photobiological principles of

therapeutic applications of laser radiation. Biochemistry (Mosc) 2004,

69:81-90.

29. Hu WP, Wang JJ, Yu CL, Lan CC, Chen GS, Yu HS: Helium-neon laser

irradiation stimulates cell proliferation through photostimulatory effects

in mitochondria. J Invest Dermatol 2007, 127:2048-2057.

30. Moriyama Y, Nguyen J, Akens M, Moriyama EH, Lilge L: In vivo effects of

low level laser therapy on inducible nitric oxide synthase. Lasers Surg

Med 2009, 41:227-231.

31. Samoilova KA, Zhevago NA, Petrishchev NN, Zimin AA: Role of nitric oxide

in the visible light-induced rapid increase of human skin

microcirculation at the local and systemic levels: II. healthy volunteers.

Photomed Laser Surg 2008, 26:443-449.

32. Yamaura M, Yao M, Yaroslavsky I, Cohen R, Smotrich M, Kochevar IE: Low

level light effects on inflammatory cytokine production by rheumatoid

arthritis synoviocytes. Lasers Surg Med 2009, 41:282-290.

33. Shiba H, Tsuda H, Kajiya M, Fujita T, Takeda K, Hino T, Kawaguchi H,

Kurihara H: Neodymium-doped yttrium-aluminium-garnet laser

irradiation abolishes the increase in interleukin-6 levels caused by

peptidoglycan through the p38 mitogen-activated protein kinase

pathway in human pulp cells. J Endod 2009, 35:373-376.

34. de Lima Mafra F, Costa MS, Albertini R, Silva JA Jr, Aimbire F: Low level

laser therapy (LLLT): attenuation of cholinergic hyperreactivity, beta(2)-

adrenergic hyporesponsiveness and TNF-alpha mRNA expression in rat

bronchi segments in E. coli lipopolysaccharide-induced airway

inflammation by a NF-kappaB dependent mechanism. Lasers Surg Med

2009, 41:68-74.

35. Aimbire F, de Oliveira Ligeiro AP, Albertini R, Correa JC, de Campos

Ladeira CB, Lyon JP, Silva JA Jr, Costa MS: Low level laser therapy (LLLT)

decreases pulmonary microvascular leakage, neutrophil influx and IL-

1beta levels in airway and lung from rat subjected to LPS-induced

inflammation. Inflammation 2008, 31:189-197.

36. Safavi SM, Kazemi B, Esmaeili M, Fallah A, Modarresi A, Mir M: Effects of

low-level He-Ne laser irradiation on the gene expression of IL-1beta,

TNF-alpha, IFN-gamma, TGF-beta, bFGF, and PDGF in rat’s gingiva. Lasers

Med Sci 2008, 23:331-335.

37. Saygun I, Karacay S, Serdar M, Ural AU, Sencimen M, Kurtis B: Effects of

laser irradiation on the release of basic fibroblast growth factor (bFGF),

insulin like growth factor-1 (IGF-1), and receptor of IGF-1 (IGFBP3) from

gingival fibroblasts. Lasers Med Sci 2008, 23:211-215.

38. Schwartz F, Brodie C, Appel E, Kazimirsky G, Shainberg A: Effect of helium/

neon laser irradiation on nerve growth factor synthesis and secretion in

skeletal muscle cultures. J Photochem Photobiol B 2002, 66:195-200.

39. Yu W, Naim JO, Lanzafame RJ: The effect of laser irradiation on the

release of bFGF from 3T3 fibroblasts. Photochem Photobiol 1994,

59:167-170.

40. Zungu IL, Hawkins Evans D, Abrahamse H: Mitochondrial responses of

normal and injured human skin fibroblasts following low level laser

irradiation–an in vitro study. Photochem Photobiol 2009, 85:987-996.

41. Wu S, Xing D, Gao X, Chen WR: High fluence low-power laser irradiation

induces mitochondrial permeability transition mediated by reactive

oxygen species. J Cell Physiol 2009, 218:603-611.

42. Lan CC, Wu CS, Chiou MH, Chiang TY, Yu HS: Low-energy helium-neon

laser induces melanocyte proliferation via interaction with type IV

collagen: visible light as a therapeutic option for vitiligo. Br J Dermatol

2009, 161:273-280.

43. Karu T: Photobiology of low-power laser effects. Health Phys 1989,

56:691-704.

44. Tiphlova O, Karu T: Role of primary photoacceptors in low-power laser

effects: action of He-Ne laser radiation on bacteriophage T4-Escherichia

coli interaction. Lasers Surg Med 1989, 9:67-69.

45. Karu TI: Mitochondrial signaling in mammalian cells activated by red and

near-IR radiation. Photochem Photobiol 2008, 84:1091-1099.

Lin et al. Journal of Translational Medicine 2010, 8:16

http://www.translational-medicine.com/content/8/1/16

46. Zhang L, Xing D, Zhu D, Chen Q: Low-power laser irradiation inhibiting

Abeta25-35-induced PC12 cell apoptosis via PKC activation. Cell Physiol

Biochem 2008, 22:215-222.

47. Aimbire F, Santos FV, Albertini R, Castro-Faria-Neto HC, Mittmann J,

Pacheco-Soares C: Low-level laser therapy decreases levels of lung

neutrophils anti-apoptotic factors by a NF-kappaB dependent

mechanism. Int Immunopharmacol 2008, 8:603-605.

48. Lipovsky A, Nitzan Y, Lubart R: A possible mechanism for visible lightinduced

wound healing. Lasers Surg Med 2008, 40:509-514.

49. Ignatieva N, Zakharkina O, Andreeva I, Sobol E, Kamensky V, Lunin V: Effects

of laser irradiation on collagen organization in chemically induced

degenerative annulus fibrosus of lumbar intervertebral disc. Lasers Surg

Med 2008, 40:422-432.

50. Silveira LB, Prates RA, Novelli MD, Marigo HA, Garrocho AA, Amorim JC,

Sousa GR, Pinotti M, Ribeiro MS: Investigation of mast cells in human

gingiva following low-intensity laser irradiation. Photomed Laser Surg

2008, 26:315-321.

51. Coombe AR, Ho CT, Darendeliler MA, Hunter N, Philips JR, Chapple CC,

Yum LW: The effects of low level laser irradiation on osteoblastic cells.

Clin Orthod Res 2001, 4:3-14.

52. Gregoraszczuk E, Dobrowolski JW, Galas J: Effect of low intensity laser

beam on steroid dehydrogenase activity and steroid hormone

production in cultured porcine granulosa cells. Folia Histochem Cytochem

(Krakow) 1983, 21:87-92.

53. Yu HS, Chang KL, Yu CL, Chen JW, Chen GS: Low-energy helium-neon

laser irradiation stimulates interleukin-1 alpha and interleukin-8 release

from cultured human keratinocytes. J Invest Dermatol 1996, 107:593-596.

54. Mvula B, Mathope T, Moore T, Abrahamse H: The effect of low level laser

irradiation on adult human adipose derived stem cells. Lasers Med Sci

2008, 23:277-282.

55. Mvula B, Moore TJ, Abrahamse H: Effect of low-level laser irradiation and

epidermal growth factor on adult human adipose-derived stem cells.

Lasers Med Sci 25:33-39.

56. Hawkins DH, Abrahamse H: The role of laser fluence in cell viability,

proliferation, and membrane integrity of wounded human skin

fibroblasts following helium-neon laser irradiation. Lasers Surg Med 2006,

38:74-83.

57. Houreld N, Abrahamse H: In vitro exposure of wounded diabetic

fibroblast cells to a helium-neon laser at 5 and 16 J/cm2. Photomed Laser

Surg 2007, 25:78-84.

58. Smol’yaninova NK, Karu TI, Fedoseeva GE, Zelenin AV: Effects of He-Ne

laser irradiation on chromatin properties and synthesis of nucleic acids

in human peripheral blood lymphocytes. Biomed Sci 1991, 2:121-126.

59. Funk JO, Kruse A, Neustock P, Kirchner H: Helium-neon laser irradiation

induces effects on cytokine production at the protein and the mRNA

level. Exp Dermatol 1993, 2:75-83.

60. Funk JO, Kruse A, Kirchner H: Cytokine production after helium-neon laser

irradiation in cultures of human peripheral blood mononuclear cells. J

Photochem Photobiol B 1992, 16:347-355.

61. Gulsoy M, Ozer GH, Bozkulak O, Tabakoglu HO, Aktas E, Deniz G, Ertan C:

The biological effects of 632.8-nm low energy He-Ne laser on peripheral

blood mononuclear cells in vitro. J Photochem Photobiol B 2006,

82:199-202.

62. Novoselova EG, Cherenkov DA, Glushkova OV, Novoselova TV,

Chudnovskii VM, Iusupov VI, Fesenko EE: [Effect of low-intensity laser

radiation (632.8 nm) on immune cells isolated from mice]. Biofizika 2006,

51:509-518.

63. Dube A, Bansal H, Gupta PK: Modulation of macrophage structure and

function by low level He-Ne laser irradiation. Photochem Photobiol Sci

2003, 2:851-855.

64. Hemvani N, Chitnis DS, Bhagwanani NS: Helium-neon and nitrogen laser

irradiation accelerates the phagocytic activity of human monocytes.

Photomed Laser Surg 2005, 23:571-574.

65. Moustsen PA, Vinter N, Aas-Andersen L, Kragstrup J: [Laser treatment of

sinusitis in general practice assessed by a double-blind controlled

study]. Ugeskr Laeger 1991, 153:2232-2234.

66. Shen X, Zhao L, Ding G, Tan M, Gao J, Wang L, Lao L: Effect of combined

laser acupuncture on knee osteoarthritis: a pilot study. Lasers Med Sci

2009, 24:129-136.

67. Ekim A, Armagan O, Tascioglu F, Oner C, Colak M: Effect of low level laser

therapy in rheumatoid arthritis patients with carpal tunnel syndrome.

Swiss Med Wkly 2007, 137:347-352.

68. Minatel DG, Frade MA, Franca SC, Enwemeka CS: Phototherapy promotes

healing of chronic diabetic leg ulcers that failed to respond to other

therapies. Lasers Surg Med 2009, 41:433-441.

69. Bocci V, Travagli V, Zanardi I: May oxygen-ozone therapy improves

cardiovascular disorders?. Cardiovasc Hematol Disord Drug Targets 2009,

9:78-85.

70. Bocci V, Borrelli E, Travagli V, Zanardi I: The ozone paradox: ozone is a

strong oxidant as well as a medical drug. Med Res Rev 2009, 29:646-682.

71. Re L, Mawsouf MN, Menendez S, Leon OS, Sanchez GM, Hernandez F:

Ozone therapy: clinical and basic evidence of its therapeutic potential.

Arch Med Res 2008, 39:17-26.

72. Damera G, Zhao H, Wang M, Smith M, Kirby C, Jester WF, Lawson JA,

Panettieri RA Jr: Ozone modulates IL-6 secretion in human airway

epithelial and smooth muscle cells. Am J Physiol Lung Cell Mol Physiol

2009, 296:L674-683.

73. Manzer R, Dinarello CA, McConville G, Mason RJ: Ozone exposure of

macrophages induces an alveolar epithelial chemokine response

through IL-1alpha. Am J Respir Cell Mol Biol 2008, 38:318-323.

74. McDonald RJ, Usachencko J: Neutrophils injure bronchial epithelium after

ozone exposure. Inflammation 1999, 23:63-73.

75. Rodriguez ZZ, Guanche D, Alvarez RG, Rosales FH, Alonso Y, Schulz S:

Preconditioning with ozone/oxygen mixture induces reversion of some

indicators of oxidative stress and prevents organic damage in rats with

fecal peritonitis. Inflamm Res 2009.

76. Zamora ZB, Borrego A, Lopez OY, Delgado R, Gonzalez R, Menendez S,

Hernandez F, Schulz S: Effects of ozone oxidative preconditioning on

TNF-alpha release and antioxidant-prooxidant intracellular balance in

mice during endotoxic shock. Mediators Inflamm 2005, 2005:16-22.

77. Borrego A, Zamora ZB, Gonzalez R, Romay C, Menendez S, Hernandez F,

Montero T, Rojas E: Protection by ozone preconditioning is mediated by

the antioxidant system in cisplatin-induced nephrotoxicity in rats.

Mediators Inflamm 2004, 13:13-19.

78. Martinez-Sanchez G, Al-Dalain SM, Menendez S, Re L, Giuliani A, Candelario-

Jalil E, Alvarez H, Fernandez-Montequin JI, Leon OS: Therapeutic efficacy of

ozone in patients with diabetic foot. Eur J Pharmacol 2005, 523:151-161.

79. Surinchak JS, Alago ML, Bellamy RF, Stuck BE, Belkin M: Effects of low-level

energy lasers on the healing of full-thickness skin defects. Lasers Surg

Med 1983, 2:267-274.

80. Broadley C, Broadley KN, Disimone G, Riensch L, Davidson JM: Low-energy

helium-neon laser irradiation and the tensile strength of incisional

wounds in the rat. Wound Repair Regen 1995, 3:512-517.

81. Ghamsari SM, Taguchi K, Abe N, Acorda JA, Yamada H: Histopathological

effect of low-level laser therapy on sutured wounds of the teat in dairy

cattle. Vet Q 1996, 18:17-21.

82. Ghamsari SM, Taguchi K, Abe N, Acorda JA, Sato M, Yamada H: Evaluation

of low level laser therapy on primary healing of experimentally induced

full thickness teat wounds in dairy cattle. Vet Surg 1997, 26:114-120.

83. Pinfildi CE, Liebano RE, Hochman BS, Ferreira LM: Helium-neon laser in

viability of random skin flap in rats. Lasers Surg Med 2005, 37:74-77.

84. Maiya GA, Kumar P, Rao L: Effect of low intensity helium-neon (He-Ne)

laser irradiation on diabetic wound healing dynamics. Photomed Laser

Surg 2005, 23:187-190.

85. Bayat M, Vasheghani MM, Razavi N, Taheri S, Rakhshan M: Effect of lowlevel

laser therapy on the healing of second-degree burns in rats: a

histological and microbiological study. J Photochem Photobiol B 2005,

78:171-177.

86. Garavello I, Baranauskas V, da Cruz-Hofling MA: The effects of low laser

irradiation on angiogenesis in injured rat tibiae. Histol Histopathol 2004,

19:43-48.

87. Lin YS, Huang MH, Chai CY, Yang RC: Effects of helium-neon laser on

levels of stress protein and arthritic histopathology in experimental

osteoarthritis. Am J Phys Med Rehabil 2004, 83:758-765.

88. Jamieson RW, Friend PJ: Organ reperfusion and preservation. Front Biosci

2008, 13:221-235.

89. Akai M, Usuba M, Maeshima T, Shirasaki Y, Yasuoka S: Laser’s effect on

bone and cartilage change induced by joint immobilization: an

experiment with animal model. Lasers Surg Med 1997, 21:480-484.

Lin et al. Journal of Translational Medicine 2010, 8:16

http://www.translational-medicine.com/content/8/1/16

90. Nakano J, Kataoka H, Sakamoto J, Origuchi T, Okita M, Yoshimura T: Lowlevel

laser irradiation promotes the recovery of atrophied gastrocnemius

skeletal muscle in rats. Exp Physiol 2009, 94:1005-1015.

91. Doin-Silva R, Baranauskas V, Rodrigues-Simioni L, da Cruz-Hofling MA: The

ability of low level laser therapy to prevent muscle tissue damage

induced by snake venom. Photochem Photobiol 2009, 85:63-69.

92. Gal D, Chokshi SK, Mosseri M, Clarke RH, Isner JM: Percutaneous delivery of

low-level laser energy reverses histamine-induced spasm in

atherosclerotic Yucatan microswine. Circulation 1992, 85:756-768.

93. Kipshidze N, Sahota H, Komorowski R, Nikolaychik V, Keelan MH Jr:

Photoremodeling of arterial wall reduces restenosis after balloon

angioplasty in an atherosclerotic rabbit model. J Am Coll Cardiol 1998,

31:1152-1157.

94. Novoselova EG, Glushkova OV, Cherenkov DA, Chudnovsky VM, Fesenko EE:

Effects of low-power laser radiation on mice immunity. Photodermatol

Photoimmunol Photomed 2006, 22:33-38.

95. Bensadoun RJ, Franquin JC, Ciais G, Darcourt V, Schubert MM, Viot M,

Dejou J, Tardieu C, Benezery K, Nguyen TD, Laudoyer Y, Dassonville O,

Poissonnet G, Vallicioni J, Thyss A, Hamdi M, Chauvel P, Demard F: Lowenergy

He/Ne laser in the prevention of radiation-induced mucositis. A

multicenter phase III randomized study in patients with head and neck

cancer. Support Care Cancer 1999, 7:244-252.

96. Arun Maiya G, Sagar MS, Fernandes D: Effect of low level helium-neon

(He-Ne) laser therapy in the prevention & treatment of radiation

induced mucositis in head & neck cancer patients. Indian J Med Res 2006,

124:399-402.

97. Neiburger EJ: Rapid healing of gingival incisions by the helium-neon

diode laser. J Mass Dent Soc 1999, 48:8-13, 40..

98. Aras MH, Gungormus M: Placebo-controlled randomized clinical trial of

the effect two different low-level laser therapies (LLLT)-intraoral and

extraoral-on trismus and facial swelling following surgical extraction of

the lower third molar. Lasers Med Sci 2009.

99. Zycinski P, Krzeminska-Pakula M, Peszynski-Drews C, Kierus A, Trzos E,

Rechcinski T, Figiel L, Kurpesa M, Plewka M, Chrzanowski L, Drozdz J: Laser

biostimulation in end-stage multivessel coronary artery disease–a

preliminary observational study. Kardiol Pol 2007, 65:13-21, discussion 22-

13..

100. Zivin JA, Albers GW, Bornstein N, Chippendale T, Dahlof B, Devlin T,

Fisher M, Hacke W, Holt W, Ilic S, Kasner S, Lew R, Nash M, Perez J,

Rymer M, Schellinger P, Schneider D, Schwab S, Veltkamp R, Walker M,

Streeter J, NeuroThera Effectiveness and Safety Trial-2 Investigators:

Effectiveness and safety of transcranial laser therapy for acute ischemic

stroke. Stroke 2009, 40:1359-1364.

101. Osiris Therapeutics Reports Interim Data for COPD Trial. http://www.

medicalnewstoday.com/articles/155267.php.

102. Gavish L, Perez LS, Reissman P, Gertz SD: Irradiation with 780 nm diode

laser attenuates inflammatory cytokines but upregulates nitric oxide in

lipopolysaccharide-stimulated macrophages: implications for the

prevention of aneurysm progression. Lasers Surg Med 2008, 40:371-378.

103. Correa F, Lopes Martins RA, Correa JC, Iversen VV, Joenson J, Bjordal JM:

Low-level laser therapy (GaAs lambda = 904 nm) reduces inflammatory

cell migration in mice with lipopolysaccharide-induced peritonitis.

Photomed Laser Surg 2007, 25:245-249.

104. Aimbire F, Lopes-Martins RA, Castro-Faria-Neto HC, Albertini R,

Chavantes MC, Pacheco MT, Leonardo PS, Iversen VV, Bjordal JM: Low-level

laser therapy can reduce lipopolysaccharide-induced contractile force

dysfunction and TNF-alpha levels in rat diaphragm muscle. Lasers Med

Sci 2006, 21:238-244.

105. de Morais NC, Barbosa AM, Vale ML, Villaverde AB, de Lima CJ, Cogo JC,

Zamuner SR: Anti-Inflammatory Effect of Low-Level Laser and Light-

Emitting Diode in Zymosan-Induced Arthritis. Photomed Laser Surg 2009.

106. Boschi ES, Leite CE, Saciura VC, Caberlon E, Lunardelli A, Bitencourt S,

Melo DA, Oliveira JR: Anti-Inflammatory effects of low-level laser therapy

(660 nm) in the early phase in carrageenan-induced pleurisy in rat.

Lasers Surg Med 2008, 40:500-508.

107. Shimizu N, Mayahara K, Kiyosaki T, Yamaguchi A, Ozawa Y, Abiko Y: Lowintensity

laser irradiation stimulates bone nodule formation via insulinlike

growth factor-I expression in rat calvarial cells. Lasers Surg Med 2007,

39:551-559.

108. Hackett TL, Shaheen F, Johnson A, Wadsworth S, Pechkovsky DV,

Jacoby DB, Kicic A, Stick SM, Knight DA: Characterization of side

population cells from human airway epithelium. Stem Cells 2008,

26:2576-2585.

109. Irwin D, Helm K, Campbell N, Imamura M, Fagan K, Harral J, Carr M,

Young KA, Klemm D, Gebb S, Dempsey EC, West J, Majka S: Neonatal lung

side population cells demonstrate endothelial potential and are altered

in response to hyperoxia-induced lung simplification. Am J Physiol Lung

Cell Mol Physiol 2007, 293:L941-951.

110. Thebaud B, Ladha F, Michelakis ED, Sawicka M, Thurston G, Eaton F,

Hashimoto K, Harry G, Haromy A, Korbutt G, Archer SL: Vascular

endothelial growth factor gene therapy increases survival, promotes

lung angiogenesis, and prevents alveolar damage in hyperoxia-induced

lung injury: evidence that angiogenesis participates in alveolarization.

Circulation 2005, 112:2477-2486.

111. Thebaud B: Angiogenesis in lung development, injury and repair:

implications for chronic lung disease of prematurity. Neonatology 2007,

91:291-297.

112. Schindl A, Merwald H, Schindl L, Kaun C, Wojta J: Direct stimulatory effect

of low-intensity 670 nm laser irradiation on human endothelial cell

proliferation. Br J Dermatol 2003, 148:334-336.

113. Kipshidze N, Nikolaychik V, Keelan MH, Shankar LR, Khanna A, Kornowski R,

Leon M, Moses J: Low-power helium: neon laser irradiation enhances

production of vascular endothelial growth factor and promotes growth

of endothelial cells in vitro. Lasers Surg Med 2001, 28:355-364.

114. Ihsan FR: Low-level laser therapy accelerates collateral circulation and

enhances microcirculation. Photomed Laser Surg 2005, 23:289-294.

115. Taguchi A, Soma T, Tanaka H, Kanda T, Nishimura H, Yoshikawa H,

Tsukamoto Y, Iso H, Fujimori Y, Stern DM, Naritomi H, Matsuyama T:

Administration of CD34+ cells after stroke enhances neurogenesis via

angiogenesis in a mouse model. J Clin Invest 2004, 114:330-338.

116. Li WT, Leu YC: Effects of low level red-light irradiation on the

proliferation of mesenchymal stem cells derived from rat bone marrow.

Conf Proc IEEE Eng Med Biol Soc 2007, 2007:5830-5833.

117. Tuby H, Maltz L, Oron U: Low-level laser irradiation (LLLI) promotes

proliferation of mesenchymal and cardiac stem cells in culture. Lasers

Surg Med 2007, 39:373-378.

118. Tuby H, Maltz L, Oron U: Implantation of low-level laser irradiated

mesenchymal stem cells into the infarcted rat heart is associated with

reduction in infarct size and enhanced angiogenesis. Photomed Laser

Surg 2009, 27:227-233.

119. Agaiby AD, Ghali LR, Wilson R, Dyson M: Laser modulation of angiogenic

factor production by T-lymphocytes. Lasers Surg Med 2000, 26:357-363.

120. Zhang H, Hou JF, Shen Y, Wang W, Wei YJ, Hu S: Low Level Laser

Irradiation Precondition to Create Friendly Milieu of Infarcted

Myocardium and Enhance Early Survival of Transplanted Bone Marrow

Cells. J Cell Mol Med 2009.

121. Hou JF, Zhang H, Yuan X, Li J, Wei YJ, Hu SS: In vitro effects of low-level

laser irradiation for bone marrow mesenchymal stem cells: proliferation,

growth factors secretion and myogenic differentiation. Lasers Surg Med

2008, 40:726-733.

doi:10.1186/1479-5876-8-16

Cite this article as: Lin et al.: Lasers, stem cells, and COPD. Journal of

Translational Medicine 2010 8:16.

Original Source: http://www.translational-medicine.com/content/8/1/16