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The photobiomodulation effect of higher-fluence 808-nm laser therapy with a flat-top handpiece on the wound healing of the earthworm Dendrobaena veneta: a brief report

Authors:
Andrea Amaroli at University of Genoa
Andrea Amaroli
Sara Ferrando at University of Genoa
Sara Ferrando
Reem Hanna
Reem Hanna
Reem Hanna
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Lorenzo Gallus at University of Genoa
Lorenzo Gallus
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BRIEF REPORT
The photobiomodulation effect of higher-fluence 808-nm laser
therapy with a flat-top handpiece on the wound healing
of the earthworm Dendrobaena veneta: a brief report
Andrea Amaroli
1,2
&Sara Ferrando
1
&Reem Hanna
2,3
&Lorenzo Gallus
1
&
Alberico Benedicenti
2
&Sonia Scarfì
1
&Marina Pozzolini
1
&Stefano Benedicenti
1
Received: 14 June 2016 /Accepted: 14 December 2016
#Springer-Verlag London 2017
Introduction
Photobiomodulation (PBM) is a useful tool employed in
wound healing [1]. Several studies in vitro indicate that
PBM with <500 mW average power and 3–10 J/cm
2
fluences
enhances the wound healing process in a wide range of con-
ditions in humans [2]. However, a lot of confusion still reigns
in this field since, to date, the correct laser parameters of de-
posited energy density that effectively promote cell rescue
without significant side effects are still elusive. To date, lim-
ited evidences suggest that the energy can be applied with the
same efficiency from cellular to organismic level [3].
Furthermore, there are issues using low-power and fluence
settings in the adoption of a standardized and worldwide use
protocol. The interaction between a tissue and energy in the
range of 600–1400 nm is based on scattering. When this type
of energy goes through a tissue, a rapid decrease in its density
can be observed [3]. Subsequently, it gets dispersed missing
the target and failing a real therapeutic effect [3].
Within this context, a novel flat-top handpiece
(AB2799) has been recently produced (LAMBDA SpA,
Italy). This particular probe provides homogeneous irradi-
ation and energy density by using relatively high power
densities and fluences with less risk of causing thermal
damage [4]. Our previous results using 808 nm with this
device at higher fluence (64 J/cm
2
)andpower(1W)in
continuous-wave (CW) showed an increase of mitochon-
drial activities such as oxygen consumption and ATP pro-
duction in the unicellular organism Paramecium
primaurelia [4–6] enhancing both its swimming speed
[4] and its fission rate rhythm [7]. In the current work,
we moved from the unicellular to the multicellular level.
Invertebrate species are as diverse as flies and worms and
share a lot of genes and molecular pathways with humans.
More than 90% of the domains that can be identified in
human proteins are present in fruit fly and worm proteins
while 61% of fruit fly proteins and 43% of worm proteins
have high sequence similarities to predicted human pro-
teins [8]. Furthermore, the gene conservation of earth-
worms is closer to humans than worms like the well-
known animal model Caenorhabditis elegans [9]. Thus,
we set up a new animal model based on the earthworm
*Stefano Benedicenti
Stefano.Benedicenti@unige.it
Andrea Amaroli
andrea.amaroli.71@gmail.com
Sara Ferrando
sara131274@gmail.com
Reem Hanna
reemhanna@hotmail.com
Lorenzo Gallus
galluslorenzo@gmail.com
Alberico Benedicenti
benedicenti@unige.it
Sonia Scarfì
soniascarfi@unige.it
Marina Pozzolini
marina.pozzolini@unige.it
1
Department of Earth, Environmental and Life Sciences, University of
Genoa, Corso Europa 26, 16132 Genoa, Italy
2
Department of Surgical Sciences and Integrated Diagnostic,
University of Genoa, Largo R. Benzi 10, 16132 Genoa, Italy
3
Department of Oral Surgery, Dental Institute, King’sCollege
Hospital NHS Foundation Trust, Denmark Hill, London SE5 9RS,
UK
Lasers Med Sci
DOI 10.1007/s10103-016-2132-3
Dendrobaena veneta to evaluate the effectiveness of the
flat-top handpiece at 808 nm on the wound healing pro-
cess of the earthworm by examining the macroscopic,
histological, and immunohistochemical changes on the ir-
radiated tail-excised earthworms with respect to controls.
Materials and methods
Experimental setting
D. veneta (Phylum: Annelida, Class: Oligochaeta) adult ma-
ture hermaphrodite earthworms were used in these exper-
iments. They were obtained from Redbug S.r.l. (Milano,
Italy), acclimated in laboratory into organic soil at the
temperature of 22 ± 1 °C and oatmeal fed. Selected spec-
imens with a similar length, weight, and color were rinsed
with water and anesthetized on an ice block for a few
minutes. Twenty caudal segments were excised from the
animals and half was used as controls while the other half
was irradiated with a 808-nm diode using a flat-top
handpiece (AB2799, Doctor Smile–LAMBDA SpA–
Vicenza, Italy), with a fluence of 64 J/cm
2
and a power
of 1 W in CW.
Macroscopic analysis
All experimental specimens were examined on a Zeiss stereo-
microscope, and images were acquired with a Cellpad E
(TiEsseLab S.r.l., Italy).
Histochemical and immunohistochemical analysis
All experimental specimens were placed in individual boxes
(Ø19 cm), with 60 g of organic soil and 50 ml of water. After
24 h of recovery, both the control and laser-treated earth-
worms were rinsed with water and placed on a wet block of
ice.
Then, 10 caudal body segments were further excised
from all animals (10 controls and 10 treated). All segments
were fixed in 4% paraformaldehyde (7.4 pH), paraffin em-
bedded, and sagittally sectioned into 5 μm-thick slides.
The hematoxylin-eosin staining was used for histological
observations while the immunohistochemistry analysis
was performed by using the anti-acetylchlinesterase
(AChE) rabbit polyclonal antibody (dilution 1:200)
(Santa Cruz Biotechnology, USA) followed by Alexa488
chicken anti-rabbit antibody (Invitrogen, USA) (dilution
1:800). The specimens were examined using a Leica
DMRB light and epifluorescence microscope. The images
were taken by a Leica CCD camera DFC420C (Leica,
Germany). Image analysis was performed by using
ImageJ v3.91 software. Each experiment was carried out
three times.
Immunoblot analysis
This analysis was performed to evaluate the specificity of the
immunorecognition of the anti-AChE rabbit polyclonal anti-
body. D. veneta protein lysates were obtained and blotted by
standard methods [10]. Band detection was performed using
the ChemiDoc MP system (Bio-Rad, USA).
Fig. 1 Diagram of experimental
design (A,B). Wound healing
process cartoon in control (A’)and
irradiated (B’)Dendrobaena
veneta specimens 24 h after
treatments
Lasers Med Sci
Results and discussion
The establishment of experimental animal models allows the
researchers to deepen their knowledge on human physiology
and diseases [11]. Recently, the use of vertebrate models has
been repeatedly challenged for ethical reasons inspiring the
setup of new invertebrate model organisms of diseases with
limited sentience. In this study, we used a new wound healing
model (Fig. 1) in the earthworm D. veneta to study the effects
of the 808-nm laser therapy.
All the earthworms survived during the experiments and
beyond and looked healthy despite the amputations,
confirming the results of a previous study on earthworm re-
generation [12].
After caudal dissections, macroscopic observations of the
wound sites (Fig. 2a) allowed us to detect all the tissues of the
body wall (epidermis, muscular layer, coelom’scells,blood
vessel, alimentary canal). As compared to control-dissected
animals, irradiated-dissected animals (Fig. 2b) immediately
after treatment showed a clear muscular contraction at the
wound site pointed out by the dorsal-ventral thinning of the
body and the internalization of the alimentary tract which in
fact disappeared from the wound site only in the irradiated
earthworms. Furthermore, at 24 h, a significant wound closure
Fig. 2 Effect of 808-nm laser therapy on wound repair in Dendrobaena
veneta.a–dMacroscopic images of amputated specimens. aNon-
irradiated control specimen. bSpecimen immediately after irradiation. c
Twenty-four-hour control specimen. dTwenty-four-hour irradiated
specimen. e–nHematoxylin-eosin-stained histological sections. e
Twenty-four-hour control specimen with evident coelomic plug. fDetail
of 24-h control specimen coelomic cavity. i,lDetails of 24-h control
specimen coelomic plug. gTwenty-four-hour irradiated specimen.
Coelomic plug is thinner than control. hDetail of 24-h control specimen
coelomic cavity. m,nDetails of the 24-h irradiated specimen plug area. o,
pAChE immunofluorescence on histological sections. oTwenty-four-
hour control specimen; intense immunoreactivity in the plug area high-
lights the non-cholinergic AChE. pTwenty-four-hour irradiated
specimen; immunoreactivity is evident in the cholinergic innervated
zones, while it is very reduced in the plug area. qImmunoblot. Lane 1:
markers. Lane 2:D. veneta homogenate. The antibody recognizes only
one band of molecular mass at 70 kDa. ac alimentary canal, ace
alimentary canal epidermis, bblood, bv blood vessel, ccoelomocytes,
cl chloragogen cell, eepidermis, mmuscle, pe peritoneum, pl coelomic
plug, sseptum. Scale bars,100μm
Lasers Med Sci
is observable only in the irradiated samples compared to con-
trols (Fig. 2d vs c). These findings seem to confirm
Shamirzaev’s[13] and Moskvin’s[14] results that showed
marked spasmolytic properties of the low-power laser irradi-
ation, likely due to Ca
2+
signaling in the adjacent muscle’s
cells. This provides a coordination of myocyte contraction
subsequently inducing the laser therapeutic effect.
The contraction of the body and the mechanical thinning of
its walls in the irradiated animals with respect to controls were
confirmed by the histological sections performed 24 h after
treatment (Fig. 2e, control, vs g, irradiated). Furthermore, 24 h
after treatments, controls showed a wide coelom delimited by
the peritoneum and by the chloragogen layer, which
enveloped the gut externally (Fig. 2e, f) while the irradiated
samples showed a slim ventral body with a compressed coe-
lom (Fig. 2g, h).
The main difference between the controls and the irradiated
samples was the formation of the coelomic plug, which was
evident in the histological sections of the control animals
(Fig. 2e, i, l and scheme in Fig. 1(A)) and significantly re-
duced in the irradiated ones (Fig. 2g, m, n and scheme in Fig. 1
(B)).
It is known that the coelomic plug is formed by passive
flow of coelomocytes and by active migration of cells from
other segments [15] as also displayed in the scheme in Fig. 1.
The formation of a smaller plug in the irradiated samples
could be due to the muscular contraction, which would slow
down the leakage of coelomic fluid and of coelomocytes from
the wound, causing an abatement of the inflammatoryprocess.
In the earthworm like other higher animals and human
species, inflammation is a complex biological response of
vascular tissues to infection caused by pathogens and/or dam-
aged cells [16]. In humans, the cholinergic system is involved
in the basic functions of the skin like proliferation and differ-
entiation of keratinocytes, cell adhesion and migration, blood
circulation, angiogenesis, and other immune reactions [16].
In the skin inflammatory process, in particular, the enzyme
AChE has shown an active non-cholinergic role [16]. The
nematode C. elegans AChE has great similarity to the human
AChE. In the Oligochaeta, through the D. veneta genome
sequencing (SRA database srx404365), a partial sequence
covering the 53% of the C-terminal epitope used for the gen-
eration of the rabbit polyclonal anti-AChE antibody can be
retrieved. Homology analysis of D. veneta partial sequence
via blastn algorithm shows a 49% identity to human AChE.
Indeed, our immunoblot analysis, using the abovementioned
antibody, seems to confirm the genome sequencing data,
showing a single polypeptide band with a coherent molecular
mass around 70 kDa (Fig. 2q).
AChE immunohistochemistry of sagittal sections obtained
24 h after treatments show the presence of AChE in the nerve
fibers on the peritoneal layer of the coelom, on the muscular
cells of the septum, and, only in the control animals, in a non-
innervated area of the wound, which corresponds to the coe-
lomic plug (Fig. 2o, control, vs p, irradiated). Indeed, the
optical density (OD) quantification of fluorescence in the area
of the coelomic plug shows a significant reduction of AChE
expression (p<0.001,by paired ttest) in the irradiated sam-
ples (673 ± 32.5 OD) as compared to the controls (2432 ± 150
OD) clearly indicating a laser anti-inflammatory effect.
In conclusion, the flat-top handpiece 808-nm phototherapy
with higher power and fluence promotes the wound healing in
the D. veneta new animal model through muscular contrac-
tion, which reduces the wound area, and via inhibition of the
inflammatory process demonstrated by the reduction of the
coelomic plug and of the non-cholinergic AChE production.
Compliance with ethical standards
Funding statement Fund from any funding agency was not procured.
Conflict of interest The authors declare that they have no conflict of
interest.
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... The gene expression profile of the inflammatory processes, apoptosis, heat stress, growth factors, collagen production, oxidative stress enzyme activity, and total prokaryotic symbiont content were analysed for this purpose. The results were compared with our previous data in unicellular organisms, animals, and humans [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26], and the molecular implication and evolutionary considerations on PBM-animal cell interaction are discussed. ...
... Our present and past data on the effect of PBM on life forms have highlighted that regeneration is a property of living matter, and regardless of its level of complexity, regeneration can be affected through 810 nm light. From this point of view, the C. reniform is data can be compared with the extensive information collected in our past work on the effect of 810 nm-1 W PBM on heterogeneous organisms from protozoa to humans [10][11][12][14][15][16][17][18][20][21][22][23][24][25]61]. ...
... Moreover, Wnt signalling, as well as the Bcl-2/Bax ratio, which interacts to coordinate cell fate, are probably the next molecular targets. When moving from Porifera to Annelida [15,16] up to vertebrate animals [19,20] and humans [22,23], our data highlight the key role of PBM in modulating inflammatory processes to support faster recovery. The consistent action-effect mode induced by 810 nm-1 W PBM suggests a continuum of mechanisms, as demonstrated by the functional conservation of key molecular events regulating cell behaviour and the regeneration process of both related and unrelated taxa across wide phylogenetic distances. ...
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Chemotrophic choice as a metabolic source of energy has characterised animal cell evolution. However, light interactions with animal cell photoacceptors that are able to increase energetic metabolism (photo-biomodulation (PBM)) have been previously described. In the present study, we cut three specimens of Chondrosia reniformis into four equal parts (12 fragments), and we irradiated the regenerating edge of six fragments with the previously characterised 810 nm near-infrared light, delivered at 1 W, 60 J/cm2, 1 W/cm2, and 60 J in a continuous-wave mode for 60 s through a flat-top hand-piece with a rounded spot-size area of 1 cm2. Six fragments were irradiated with 0 W for 60 s as the controls. We performed irradiation at the time 0 h and every 24 h for a total of five administrations. We monitored the regeneration process for five days (120 h) in aquaria by examining the macroscopic and histological changes. We analysed the gene expression profile of the inflammatory processes, apoptosis, heat stress, growth factors, and collagen production and determined oxidative stress enzyme activity and the total prokaryotic symbiont content. PBM sped up C. reniformis regeneration when compared to the controls. Particularly, transforming growth factor TGF3 and TGF6 upregulation during the early phase of regeneration and TGF5 upregulation 120 h postinjury in the irradiated samples supports the positive effect of PBM in sponge tissue recovery. Conversely, the expression of TGF4, a sponge fibroblast growth factor homologue, was not affected by irradiation, indicating that multiple, independent pathways regulate the TGF genes. The results are consistent with our previous data on a wide range of organisms and humans, suggesting that PBM interaction with primary and secondary cell targets has been conserved through the evolution of life forms.
View
... To improve the clinical knowledge and the support of photobiomodulation in the treatment of aphtha healing and the standardization of the therapies, we thus set up a randomized double-blind controlled trial. In our previous works, we showed that a higherpower and -fluence laser therapy, such as 808-nm, 1 W, 1 W/cm 2 , 60 J/cm 2 , positively affects mitochondrial energetic metabolism [32][33][34], improving healing [35][36][37][38] in in vitro and preclinical studies. Plus, we characterized a novel technology based on irradiation with a flat-top beam profile that can be supportive of the effectiveness and standardization of photobiomodulation therapy [32,39] and Section 2.3 below. ...
... In accordance with our previous in vitro and preclinical studies [35][36][37][38], photobiomodulation therapy was administered through the power of 1 W irradiated in continuous wave mode (CW) for an exposure time of 60 s and on a spot size of 1 cm 2 , which allowed generating a power density of 1 W/cm 2 and a fluence of 60 J/cm 2 (energy administered = 60 J). Because the aphtha's area is wider than the laser spot size, irradiation on two points was necessary to cover the entire surface. ...
... In accordance with our previous works [35][36][37][38], the predictor variable was the photobiomodulation through higher energy and power irradiated through the flat-top handpiece. ...
808-Nm Near-Infrared Laser Photobiomodulation versus Switched-Off Laser Placebo in Major Aphthae Management: A Randomized Double-Blind Controlled Trial
Article
Full-text available
  • May 2021
Background: the aphtha is one of the most common oral mucosal ulcerations and presents as a painful punched-out sore. Systemic and topical medications are used to reduce inflammation and pain and to support the natural period of remission. Alternative treatment modalities have been requested to relieve pain and improve its healing. In this regard, photobiomodulation, which is a manipulation of cells’ metabolism through an energy transfer by light sources of non-ablative or thermal intensity, could support aphtha management. The predictor variable of our research was the photobiomodulation through higher energy and power irradiated through a handpiece with a flat-top beam profile. The primary end point was the complete healing of the aphtha, defined as the time from the irradiation to the complete recovery. The secondary end point was pain relief, evaluated daily through the visual analogue scale (VAS), from the irradiation to 24 and 48 h after. Methods: a randomized, double-blind, controlled trial was conducted according to the CONSORT guideline. Irradiation was performed through an 808-nm diode laser with flat-top handpiece, and 1 W, 1 W/cm2, 60 J, 60 J/cm2 for 60 s on a spot-size area of 1 cm2. Time of complete healing and pain evaluation by VAS scale were evaluated. Results: between 1 January, 2020 and 1 March, 2021, 126 patients were screened for the study at the Department of Surgical and Diagnostic Sciences, University of Genoa, Italy. Sixty patients were randomly assigned (30 in the photobiomodulation group and 30 in the placebo group). Patients of the photobiomodulation group experienced complete healing in an average time of 8.13 days ± 1.69 (min 5–max 10 days), while for the placebo group the average time extended to 30.76 ± 4.63 days (min 25–max 42 days). Patients of the photobiomodulation therapy group experienced a statistically significant reduction in pain and discomfort 24 and 48 h after treatment (p < 0.05); the reduction was statistically higher (p < 0.05) 48 h after treatment compared to 24 h after. Conclusions: photobiomodulation at the parameters and modality of irradiation proposed accelerates the healing recovery and reduces pain compared to the patients treated with the placebo.
View
... Anti-inflammatory: attenuates cytokine storm at multiple levels and reduces the major inflammatory metabolites [189][190][191][192][193][194] Increases collagen and protein production, cell proliferation, normalized of impaired circulation, decreases edema and swelling, and improves quality and tensile strength of tissue [189][190][191][192][193][194] PBMT reduces pulmonary microvascular leakage, IL-1β, IL-6, and intracellular ROS. On this note, PBMT as a single or adjunct treatment modality can modulate the cytokine storm and ARDS via its anti-inflammatory action [196][197][198][199] Reduces inflammation and promotes lung healing 199,205,206 and minimizes the scarring process [189][190][191][192][193][194] Has an immunomodulator effect in improving immune functional and modulating the cellular and molecular activities of the cytokine storm of SARS-CoV-2 200,203,206 It is a safe, effective, low-cost modality without any side effects that may be combined with conventional treatment of ARDS [196][197][198][199] Minimizes the length of time needed on a ventilator, enhance the healing process, and shorten recovery time in COVID-19 patients 199,200 PBMT as a single or adjunct treatment modality can modulate the cytokine storm and ARDS via its anti-inflammatory action [196][197][198][199] Antioxidant: reduces pain related to inflammation via dose-dependent reduction of prostaglandin E2, prostaglandin-endoperoxide synthase-2, IL-1, IL-6, TNFa, as well as the cellular influx of neutrophils, oxidative stress [201][202][203] Prevents acidosis and hypoxia in COVID-19 patients results in SpO 2 improvement and lung healing in COVID-19 pneumonia 199,205,206,210 PBMT combined with conventional medical therapy has a potential to prevent COVID-19 progress and improve symptoms 200 PBMT combined with conventional treatment in patients with severe COVID-19 and morbid obesity is safe and effective 211 and is potential to prevent COVID-19 progress and improve symptoms 200 None encountered to date (Continued) patients. ...
... Anti-inflammatory: attenuates cytokine storm at multiple levels and reduces the major inflammatory metabolites [189][190][191][192][193][194] Increases collagen and protein production, cell proliferation, normalized of impaired circulation, decreases edema and swelling, and improves quality and tensile strength of tissue [189][190][191][192][193][194] PBMT reduces pulmonary microvascular leakage, IL-1β, IL-6, and intracellular ROS. On this note, PBMT as a single or adjunct treatment modality can modulate the cytokine storm and ARDS via its anti-inflammatory action [196][197][198][199] Reduces inflammation and promotes lung healing 199,205,206 and minimizes the scarring process [189][190][191][192][193][194] Has an immunomodulator effect in improving immune functional and modulating the cellular and molecular activities of the cytokine storm of SARS-CoV-2 200,203,206 It is a safe, effective, low-cost modality without any side effects that may be combined with conventional treatment of ARDS [196][197][198][199] Minimizes the length of time needed on a ventilator, enhance the healing process, and shorten recovery time in COVID-19 patients 199,200 PBMT as a single or adjunct treatment modality can modulate the cytokine storm and ARDS via its anti-inflammatory action [196][197][198][199] Antioxidant: reduces pain related to inflammation via dose-dependent reduction of prostaglandin E2, prostaglandin-endoperoxide synthase-2, IL-1, IL-6, TNFa, as well as the cellular influx of neutrophils, oxidative stress [201][202][203] Prevents acidosis and hypoxia in COVID-19 patients results in SpO 2 improvement and lung healing in COVID-19 pneumonia 199,205,206,210 PBMT combined with conventional medical therapy has a potential to prevent COVID-19 progress and improve symptoms 200 PBMT combined with conventional treatment in patients with severe COVID-19 and morbid obesity is safe and effective 211 and is potential to prevent COVID-19 progress and improve symptoms 200 None encountered to date (Continued) patients. ...
... Anti-inflammatory: attenuates cytokine storm at multiple levels and reduces the major inflammatory metabolites [189][190][191][192][193][194] Increases collagen and protein production, cell proliferation, normalized of impaired circulation, decreases edema and swelling, and improves quality and tensile strength of tissue [189][190][191][192][193][194] PBMT reduces pulmonary microvascular leakage, IL-1β, IL-6, and intracellular ROS. On this note, PBMT as a single or adjunct treatment modality can modulate the cytokine storm and ARDS via its anti-inflammatory action [196][197][198][199] Reduces inflammation and promotes lung healing 199,205,206 and minimizes the scarring process [189][190][191][192][193][194] Has an immunomodulator effect in improving immune functional and modulating the cellular and molecular activities of the cytokine storm of SARS-CoV-2 200,203,206 It is a safe, effective, low-cost modality without any side effects that may be combined with conventional treatment of ARDS [196][197][198][199] Minimizes the length of time needed on a ventilator, enhance the healing process, and shorten recovery time in COVID-19 patients 199,200 PBMT as a single or adjunct treatment modality can modulate the cytokine storm and ARDS via its anti-inflammatory action [196][197][198][199] Antioxidant: reduces pain related to inflammation via dose-dependent reduction of prostaglandin E2, prostaglandin-endoperoxide synthase-2, IL-1, IL-6, TNFa, as well as the cellular influx of neutrophils, oxidative stress [201][202][203] Prevents acidosis and hypoxia in COVID-19 patients results in SpO 2 improvement and lung healing in COVID-19 pneumonia 199,205,206,210 PBMT combined with conventional medical therapy has a potential to prevent COVID-19 progress and improve symptoms 200 PBMT combined with conventional treatment in patients with severe COVID-19 and morbid obesity is safe and effective 211 and is potential to prevent COVID-19 progress and improve symptoms 200 None encountered to date (Continued) patients. Over a period of 10 days, the patients were randomly assigned in a 2:1 ratio to intravenous remdesivir. ...
Understanding COVID-19 Pandemic: Molecular Mechanisms and Potential Therapeutic Strategies. An Evidence-Based Review
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Full-text available
  • Jan 2021
Initially, the SARS-CoV-2 virus was considered as a pneumonia virus; however, a series of peer reviewed medical papers published in the last eight months suggest that this virus attacks the brain, heart, intestine, nervous and vascular systems, as well the blood stream. Although many facts remain unknown, an objective appraisal of the current scientific literature addressing the latest progress on COVID-19 is required. The aim of the present study was to conduct a critical review of the literature, focusing on the current molecular structure of SARS-CoV-2 and prospective treatment modalities of COVID-19. The main objectives were to collect, scrutinize and objectively evaluate the current scientific evidence-based information, as well to provide an updated overview of the topic that is ongoing. The authors underlined potential prospective therapies, including vaccine and phototherapy, as a monotherapy or combined with current treatment modalities. The authors concluded that this review has produced high quality evidence, which can be utilized by the clinical scientific community for future reference, as the knowledge and understanding of the SARS-CoV-2 virus are evolving, in terms of its epidemiological, pathogenicity, and clinical manifestations, which ultimately map the strategic path, towards an effective and safe treatment and production of a reliable and potent vaccine.
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... It should be borne in mind that each laser wavelength carries some risk of causing collateral thermal damage which varies according to the depth of penetration of the different laser wavelengths (8,9,14). Diode laser therapy results in a peripheral zone of photobiomodulation which has anti-inflammatory, analgesic and regenerative properties (20)(21)(22). Moreover, since the laser wound heals by secondary intention, unlike a surgical excision with conventional method, there would be no need for suture or periodontal pack placement, thus improving patient comfort. ...
... The authors have ranked the different laser systems and treatment models available for dental clinicians in which adjunctive diode laser therapy to SRP has shown superior long-term (6 months) results in terms of CAL gain, in comparison with all the other laserbased treatment strategies (24). The outcomes of the present clinical report are in accordance with the inferences of all the abovementioned scientific evidence (14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24). Nevertheless, it is evident that the application of low power settings is advantageous in order to obtain long-term stability of the treatment outcome. ...
Utilisation of 810 nm Diode Laser Treatment in Periodontitis as an Alternative to Surgical Debridement Approach
Article
Full-text available
  • Mar 2021
  • PHOTOCHEM PHOTOBIOL
The efficacy of surgical periodontal may be compromised by inadequate patients’ self‐ performed plaque control or lack of supportive periodontal therapy. This clinical case report aimed to evaluate the effectiveness of 810 nm diode laser‐assisted full mouth sulcular debridement, as a potential treatment modality in the management of chronic periodontitis along with a 12‐months follow‐up evaluation. Each periodontal pocket was lased within 2 mm of deepest point, using a light contact of ‘hot tip’ of the fibre with tissue [Average power‐1W, emission mode‐continuous wave, in ‘contact’, beam diameter‐400 µm, spot area at tissue‐0.0013 cm2, fluence‐124.9 J/cm2, power density‐796 W/cm2, total energy‐ 300 J, total treatment duration‐300 seconds (5 minutes)]. The patient was followed up to 12‐months and did not report any immediate or delayed complications such as; any sign of photothermal damage, pain, tissue swelling and deformation, bleeding and infection. 810 nm diode laser‐assisted sulcular debridement at low power settings showed overall clinical and radiographic improvements and can be deemed as a potential alternative to surgical debridement approach, in persistent moderate periodontal pockets (4‐6 mm).
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... To achieve standardization and reproducibility of PBM dosimetry, a collimated flattop beam profile was employed. The flat-top beam profile allowed a static approach to treatment for consistent dosimetry and depth penetration as compared to a gaussian beam profile, which may have difficulty achieving consistent outcomes, optical delivery to target depth, as well as calculation of dosimetry [35,[37][38][39]. This is related to the unique property that the collimated flat-top, offering equal and uniform distribution of the photonic energy over more than 1 cm 2 compared with a gaussian beam profile, offering its maximum energy only at the center of the beam [37]. ...
Efficacy of 1064 nm Photobiomodulation Dosimetry Delivered with a Collimated Flat-Top Handpiece in the Management of Peripheral Facial Paralysis in Patients Unresponsive to Standard Treatment Care: A Case Series
Article
Full-text available
  • Sep 2023
Peripheral facial paralysis (PFP) is a common condition where oxidative stress (OS) is involved in the pathophysiology of facial paralysis, inhibiting peripheral nerve regeneration, which can be featured in Bell's palsy, Ramsay Hunt syndrome and Lyme disease. The current standard care treatments lack consensus and clear guidelines. Hence, the utilization of the antioxidant im-munomodulator photobiomodulation (PBM) can optimize clinical outcomes in patients who are unresponsive to standard care treatments. Our study describes three unique cases of chronic PFP of various origins that were unresponsive to standard care treatments, but achieved a significant and complete recovery of facial paralysis following PBM therapy. Case presentations: Case #1: a 30-year-old male who presented with a history of 12 years of left-side facial paralysis and tingling as a result of Bell's palsy, where all the standard care treatments failed to restore the facial muscles' paralysis. Eleven trigger and affected points were irradiated with 1064 nm with an irradiance of 0.5 W/cm 2 delivered with a collimated prototype flat-top (6 cm 2) in a pulsed mode, with a 100 µs pulse duration at a frequency of 10 Hz for 60 s (s) per point. Each point received a fluence of 30 J/cm 2 according to the following treatment protocol: three times a week for the first three months, then twice a week for another three weeks, and finally once a week for the following three months. The results showed an improvement in facial muscles' functionality (FMF) by week two, whereas significant improvement was observed after 11 weeks of PBM, after which the House-Brackmann grading scale (HBGS) of facial nerve palsy dropped to 8 from 13 prior to the treatment. Six months after PBM commencement, electromyography (EMG) showed sustainability of the FMF. Case #2: A five-year-old female who presented with a 6-month history of severe facial paralysis due to Lyme disease. The same PBM parameters were utilized, but the treatment protocol was as follows: three times a week for one month (12 consecutive treatment sessions), then the patient received seven more sessions twice a week. During the same time period, the physiotherapy of the face muscles was also delivered intensively twice a week (10 consecutive treatments in five weeks). Significant improvements in FMF and sustainability over a 6-month follow-up were observed. Case #3: A 52-year-old male who presented with severe facial palsy (Grade 6 on HBGS) and was diagnosed with Ramsay Hunt syndrome. The same laser parameters were employed, but the treatment protocol was as follows: three times a week for three weeks, then reduced to twice a week for another three weeks, then weekly for the next three months. By week 12, the patient showed a significant FMF improvement, and by week 20, complete FMF had been restored. Our results, for the first time, showed pulsed 1064 nm PBM delivered with a flat-top handpiece protocol is a valid and its treatment protocol modified, depending on the origin and severity of the condition, which is fundamental in optimizing facial paralysis recovery and alleviating neurological symptoms. Further extensive studies with large data are warranted to validate our PBM dosimetry and treatment protocols.
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... In fact, we previously showed the ability of 808 nm 1 W, 1 W/cm 2 and 60 J/cm 2 to modulate the calcium homeostasis affecting mitochondria bioenergy production [26,27], through the mitochondrial respiratory chain [23,25,55,56], cell metabolism [26,50], and inflammatory cell pathways [57], to promote cell proliferation [50,55,57], tissue regeneration [28,30,58], and release of neurotransmitters [59]. Lastly, the target of PBM therapy and the alteration of energy category genes in both moderate and severe BP could also explain why our PBM therapy was unsuccessful if performed more than 1 year after the onset of palsy. ...
Recovery from Idiopathic Facial Paralysis (Bell’s Palsy) Using Photobiomodulation in Patients Non-Responsive to Standard Treatment: A Case Series Study
Article
Full-text available
  • Aug 2021
Diminished facial movement and marked facial asymmetry can lead to a consistent psychological burden. Bell′s palsy (BP) is one of the most common causes of facial nerve illness, which comes with unilateral acute facial paresis. Nowadays, no clear guidelines for treating BP are available. We carried out a case series study to test the efficacy of photobiomodulation (PBM) therapy in patients with BP non-responsive to standard treatment. The study was experimentally performed at the Department of Surgical and Diagnostic Sciences, University of Genoa (Genoa, Italy), in accordance with case report guidelines. Patients were referred to our department by colleagues for evaluation to be included in the case series because no consistent improvement was observed at least 3 months from the diagnosis of BP. All the patients interrupted their pharmacological therapy before the initiation of PBM therapy. PBM therapy (808 nm, 1 W irradiated in continuous-wave for 60 s on spot-size 1 cm2; 1 W/cm2; 60 J/cm2; and 60 J) was administered every 2 days until complete resolution. Evaluation of the House-Brackmann scale was performed before and after treatments. Fourteen patients were screened as eligible for the study. Patients were Caucasians (36% females and 64% males) with a mean age ± standard deviation of 56.07 ± 15.21 years. Eleven patients out of 14, who experienced BP a maximum of 6 months, completely recovered through PBM. The three patients that did not show improvement were those who had experienced BP for years. PBM could be a supportive therapy for the management of BP in patients non-responsive to standard treatment. However, randomized controlled trials are necessary to sustain our encouraging results, exclude bias, and better explain the boundary between the time from diagnosis and the recovery of BP through PBM therapy.
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... It is effective in reducing inflammatory cytokines through PBM cellular signaling and Ca 2+ sensitivity, which accounts for its anti-inflammatory effects [32,34]. Additionally, studies have shown PBM effectiveness in alleviating pain [35][36][37], reducing oxidative stress and accelerating wound healing [38][39][40]. ...
A Novel Approach of Combining Methylene Blue Photodynamic Inactivation, Photobiomodulation and Oral Ingested Methylene Blue in COVID-19 Management: A Pilot Clinical Study with 12-Month Follow-Up
Article
Full-text available
  • Nov 2022
Coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 virus was first recognized in late 2019 and remains a significant threat. We therefore assessed the use of local methylene blue photodynamic viral inactivation (MB-PDI) in the oral and nasal cavities, in combination with the systemic anti-viral, anti-inflammatory and antioxidant actions of orally ingested methylene blue (MB) and photobiomodulation (PBM) for COVID-19 disease. The proposed protocol leverages the separate and combined effects of MB and 660nm red light emitted diode (LED) to comprehensively address the pathophysiological sequelae of COVID-19. A total of eight pilot subjects with COVID-19 disease were treated in the Bahamas over the period June 2021–August 2021, using a remote care program that was developed for this purpose. Although not a pre-requisite for inclusion, none of the subjects had received any COVID-19 vaccination prior to commencing the study. Clinical outcome assessment tools included serial cycle threshold measurements as a surrogate estimate of viral load; serial online questionnaires to document symptom response and adverse effects; and a one-year follow-up survey to assess long-term outcomes. All subjects received MB-PDI to target the main sites of viral entry in the nose and mouth. This was the central component of the treatment protocol with the addition of orally ingested MB and/or PBM based on clinical requirements. The mucosal surfaces were irradiated with 660 nm LED in a continuous emission mode at energy density of 49 J/cm2 for PDI and 4.9 J/cm2 for PBM. Although our pilot subjects had significant co-morbidities, extremely high viral loads and moderately severe symptoms during the Delta phase of the pandemic, the response to treatment was highly encouraging. Rapid reductions in viral loads were observed and negative PCR tests were documented within a median of 4 days. These laboratory findings occurred in parallel with significant clinical improvement, mostly within 12–24 h of commencing the treatment protocol. There were no significant adverse effects and none of the subjects who completed the protocol required in-patient hospitalization. The outcomes were similarly encouraging at one-year follow-up with virtual absence of “long COVID” symptoms or of COVID-19 re-infection. Our results indicate that the protocols may be a safe and promising approach to challenging COVID-19 disease. Moreover, due its broad spectrum of activity, this approach has the potential to address the prevailing and future COVID-19 variants and other infections transmitted via the upper respiratory tract. Extensive studies with a large cohort are warranted to validate our results.
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... Recently, new experimental organisms, earthworm species, have been introduced to investigate wound healing [13,14] and segment regeneration [6]. In our previous study, a very pronounced effect of illumination of earthworms by THz pulses was found on the regeneration of ablated tail segments in Eisenia andrei earthworms [6]. ...
Absorption of Pulsed Terahertz and Optical Radiation in Earthworm Tissue and Its Heating Effect
Article
Full-text available
  • Nov 2021
  • J INFRARED MILLIM TE
The transmission of THz, near-infrared (1030 nm), and green (515 nm) pulses through Eisenia andrei body wall is studied, which consists of epithelial layer and circular and longitudinal muscles. Samples with the full-body cross-section were also investigated. The transmitted power for the green pulses followed the Beer-Lambert law of exponential attenuation for all thicknesses and tissue structures. Different body wall and body center absorption coefficients were found in case of infrared pulses. In the THz range, the body wall absorption coefficient steadily increases from about 80 cm –1 at 0.2 THz to about 273 cm –1 at 2.5 THz. Numerical estimation indicates that THz pulses of 5-μJ energy and 1-kHz repetition rate (5-mW average power) cause only a small temperature increase of about 0.4 K, suggesting that heating has minor contribution to biological effectiveness.
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Photobiomodulation on isolated mitochondria at 810 nm: first results on the efficiency of the energy conversion process
Article
Full-text available
  • May 2024
In this paper the photobiomodulation on isolated mitochondria of bovine liver is studied as a thermodynamic process of conversion of energy. This analysis is conducted by considering a particular set-up for the photobiomodulation experiments of interest. It allows, in particular, the computation of the electromagnetic field and the related energetic quantities in the stimulated organelles. The measurements of the excess of biochemical power density produced by the illuminated mitochondria are performed at regular time intervals after the experiments. The calculations and the measurements finally allow us to obtain the first results on the efficiency of the process of conversion of electromagnetic energy into excess of biochemical energy released by the isolated organelles.
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Photobiomodulation Therapy for Management of Oral Complications-Induced by Head and Neck Cancer Treatments
Chapter
Full-text available
  • Feb 2023
This chapter covers photobiomodulation (PBM) mechanism of action, benefits and clinical applications. Also, it mentions the current clinical PBM protocols in the management of various oral complications-induced by chemotherapy and radiotherapy, in terms of therapeutic and preventive approaches. Moreover, emphasising the vital role of Dental hygienists and therapists (D&Hs) in assisting oral complications management within a multidisciplinary team at secondary care and in primary care setup. Finally, the future perspective scope of DH&Ts training in the field of laser in dentistry in general with focus on PBM therapy. Also, my chapter highlights the educational courses that H&Ts can undertake to enhance their skills and knowledge in PBM .
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808-nm laser therapy with a flat-top handpiece photobiomodulates mitochondria activities of Paramecium primaurelia (Protozoa)
Article
Full-text available
  • May 2016
  • LASER MED SCI
Photobiomodulation is proposed as a non-linear process, and only low-level laser therapy (LLLT) is assumed to stimulate exposed cells, whereas high powered laser and fluences can cause negative effects, exhausting the cell's energy reserve as a consequence of excessive photon-based stimulation. In our work, we investigated and compared the effects of 808-nm diode laser (CW) with a new flat-top handpiece. To this purpose, we tested the photobiomodulation effects of 1 and 3 J/cm(2) fluence, both generated by 100 mW or 1 W of laser power and of 64 J/cm(2) of fluence generated by 100 mW, 1 W, 1.5 W or 2 W, as expressed through oxygen consumption and ATP synthesis of Paramecium. Data collected indicates the incremental consumption of oxygen through irradiation with 3 J/cm(2)-100 mW or 64 J/cm(2)-1 W correlates with an increase in Paramecium ATP synthesis. The Paramecium respiration was inhibited by fluences 64 J/cm(2)-100 mW or 64 J/cm(2)-2 W and was followed by a decrease in the endogenous ATP concentration. The 1 J/cm(2)-100 mW or 1 W and 3 J/cm(2)-1 W did not affect mitochondrial activity. The results show that the fluence of 64 J/cm(2)-1 W more than the 3 J/cm(2)-100 mW causes greater efficiency in Paramecium mitochondria respiratory chain activity. Our results suggest that thanks to flat-top handpiece we used, high fluences by high-powered laser have to be reconsidered as an effective and non-invasive therapy. Possible associated benefits of deeper tissue penetration would increase treatment effectiveness and reduced irradiation time.
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The Protozoan, Paramecium primaurelia, as a Non-sentient Model to Test Laser Light Irradiation: The Effects of an 808nm Infrared Laser Diode on Cellular Respiration
Article
Full-text available
  • Jul 2015
Photobiomodulation (PBM) has been used in clinical practice for more than 40 years. Unfortunately, conflicting literature has led to the labelling of PBM as a complementary or alternative medicine approach. However, past and ongoing clinical and research studies by reputable investigators have re-established the merits of PBM as a genuine medical therapy, and the technique has, in the last decade, seen an exponential increase in the numbers of clinical instruments available, and their applications. This resurgence has led to a clear need for appropriate experimental models to test the burgeoning laser technology being developed for medical applications. In this context, an ethical model that employs the protozoan, Paramecium primaurelia, is proposed. We studied the possibility of using the measure of oxygen consumption to test PBM by irradiation with an infrared or nearinfrared laser. The results show that an 808nm infrared laser diode (1W; 64J/cm2) affects cellular respiration in P. primaurelia, inducing, in the irradiated cells, a significantly (p < 0.05) increased oxygen consumption of about 40%. Our findings indicate that Paramecium can be an excellent tool in biological assays involving infrared and near-infrared PBM, as it combines the advantages of in vivo results with the practicality of in vitro testing. This test represents a fast, inexpensive and straightforward assay, which offers an alternative to both traditional in vivo testing and moreexpensive mammalian cellular cultures.
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Functional properties of laser effects on morphology of liver, gall bladder and bile ducts in cholelithiasis
Article
Full-text available
  • Mar 2012
In 85 patients with calculous cholecystitis the preoperative preparation before laparoscopic cholecystectomy included irradiation of the area of gall bladder and epigastric puncture with low power magneto-infrared laser. The investigations performed revealed significant reduction of the liver and gall bladder changes both on the light optic and electron microscopic levels due to effects of low power laser irradiation.
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Mechanisms of low level light therapy
Article
Full-text available
  • Mar 2006
  • Proceedings of SPIE
The use of low levels of visible or near infrared light for reducing pain, inflammation and edema, promoting healing of wounds, deeper tissues and nerves, and preventing tissue damage has been known for almost forty years since the invention of lasers. Originally thought to be a peculiar property of laser light (soft or cold lasers), the subject has now broadened to include photobiomodulation and photobiostimulation using non-coherent light. Despite many reports of positive findings from experiments conducted in vitro, in animal models and in randomized controlled clinical trials, LLLT remains controversial. This likely is due to two main reasons; firstly the biochemical mechanisms underlying the positive effects are incompletely understood, and secondly the complexity of rationally choosing amongst a large number of illumination parameters such as wavelength, fluence, power density, pulse structure and treatment timing has led to the publication of a number of negative studies as well as many positive ones. In particular a biphasic dose response has been frequently observed where low levels of light have a much better effect than higher levels. This introductory review will cover some of the proposed cellular chromophores responsible for the effect of visible light on mammalian cells, including cytochrome c oxidase (with absorption peaks in the near infrared) and photoactive porphyrins. Mitochondria are thought to be a likely site for the initial effects of light, leading to increased ATP production, modulation of reactive oxygen species and induction of transcription factors. These effects in turn lead to increased cell proliferation and migration (particularly by fibroblasts), modulation in levels of cytokines, growth factors and inflammatory mediators, and increased tissue oxygenation. The results of these biochemical and cellular changes in animals and patients include such benefits as increased healing in chronic wounds, improvements in sports injuries and carpal tunnel syndrome, pain reduction in arthritis and neuropathies, and amelioration of damage after heart attacks, stroke, nerve injury and retinal toxicity.
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The Protozoan, Paramecium primaurelia, as a Non-sentient Model to Test Laser Light Irradiation: The Effects of an 808nm Infrared Laser Diode on Cellular Respiration
Data
  • Jul 2015
Photobiomodulation (PBM) has been used in clinical practice for more than 40 years. Unfortunately, conflicting literature has led to the labelling of PBM as a complementary or alternative medicine approach. However, past and ongoing clinical and research studies by reputable investigators have re-established the merits of PBM as a genuine medical therapy, and the technique has, in the last decade, seen an exponential increase in the numbers of clinical instruments available, and their applications. This resurgence has led to a clear need for appropriate experimental models to test the burgeoning laser technology being developed for medical applications. In this context, an ethical model that employs the protozoan, Paramecium primaurelia, is proposed. We studied the possibility of using the measure of oxygen consumption to test PBM by irradiation with an infrared or near-infrared laser. The results show that an 808nm infrared laser diode (1W; 64J/cm2) affects cellular respiration in P. primaurelia, inducing, in the irradiated cells, a significantly (p < 0.05) increased oxygen consumption of about 40%. Our findings indicate that Paramecium can be an excellent tool in biological assays involving infrared and near-infrared PBM, as it combines the advantages of in vivo results with the practicality of in vitro testing. This test represents a fast, inexpensive and straightforward assay, which offers an alternative to both traditional in vivo testing and more expensive mammalian cellular cultures. 2015 FRAME.
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Effect of 808 nm Diode Laser on Swimming Behavior, Food Vacuole Formation and Endogenous ATP Production of Paramecium primaurelia (Protozoa)
Article
  • Jun 2015
Photobiomodulation (PBM) has been used in clinical practice for more than 40 years. To clarify the mechanisms of action of PBM at cellular and organism levels, we investigated its effect on Paramecium primaurelia (Protozoa) irradiated by a 808 nm infrared diode laser with a flat-top handpiece (1 W in CW). Our results lead to the conclusion that: 1) The 808 nm laser stimulates the P. primaurelia without a thermal effect. 2) The laser effect is demonstrated by an increase in swimming speed and in food vacuole formation. 3) The laser treatment affects endogenous adenosine triphosphate (ATP) production in a positive way. 4) The effects of irradiation dose suggest an optimum exposure time of 50 s (64 J/cm(2) of fluence) to stimulate the Paramecium cells; irradiation of 25 s shows no effect or only mild effects and irradiation up to 100 s does not increase the effect observed with 50 s of treatment. 5) The increment of endogenous ATP concentration highlight the positive photobiomodulating effect of the 808 nm laser and the optimal irradiation conditions by the flat-top handpiece. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
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Paramecium: A Promising Non-Animal Bioassay to Study the Effect of 808 nm Infrared Diode Laser Photobiomodulation
Article
  • Jan 2015
Objective: Photobiostimulation and photobiomodulation (PBM) are terms applied to the manipulation of cellular behavior using low intensity light sources, which works on the principle of inducing a biological response through energy transfer. The aim of this investigation was to identify a laboratory assay to test the effect of an infrared diode laser light (808 nm) on cell fission rate. Materials and methods: Sixty cells of Paramecium primaurelia were divided in two groups of 30. The first group (test group) was irradiated, at a temperature of 24°C, for 50 sec by a 808 nm diode laser with a flat top handpiece [1 cm of spot diameter, 1 W in continuous wave (CW), 50 sec irradiation time, 64 J/cm(2) of fluence]. The second group (control group) received no laser irradiation. All cells were transferred onto a depression slide, fed, and incubated in a moist chamber at a temperature of 24°C. The cells were exposed and monitored for 10 consecutive fission rates. Changes in temperature and pH were also evaluated. Results: The exposed cells had a fission rate rhythm faster than the control cells, showing a binary fission significantly (p<0.05) shorter than unexposed cells. No significant effects of laser irradiation on pH and temperature of Paramecium's lettuce infusion medium were observed. Conclusions: The 808 nm infrared diode laser light, at the irradiation parameters used in our work, results in a precocious fission rate in P. primaurelia cells, probably through an increase in metabolic activity, secondary to an energy transfer.
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The draft sequences - Comparing species
Article
  • Feb 2001
Comparing the human genome sequences with those of other species will not only reveal what makes us genetically different. It may also help us understand what our genes do.
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Wound healing in Eisenia foetida (Oligochaeta)
Article
  • Jan 1974
Both anterior wounds (first eight segments removed) and shallow body wall wounds in Eisenia foetida are definitively closed by migration of epidermal columnar cells which undergo few morphological changes in the process. Epidermal basal cells do not contribute directly to wound epithelialization, but they enter the plug of cells which acts as the substratum for the columnar cells and function mainly as phagocytes. Epidermal gland cells, which also do not contribute to wound covering, degenerate and are phagocytized at the wound margins. The gland cells which re-form in the new wound epithelium probably arise from columnar cells. There is no change in the epidermal basement lamella during healing, but epidermal cuticle synthesis is stimulated. In the outer region of the forming cuticle the fiber bundles become disorganized and in the inner region fine filaments and larger, faintly striated fibers are laid down. The number of columnar cell microvilli, which may be involved in cuticle organization, increases after injury.
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