Acta Scientiae Veterinariae, 2021. 49(Suppl 1): 707.

CASE REPORT ISSN 1679-9216

Pub. 707

Antimicrobial Photodinamic Therapy Combined with Laser Photobiomodulation

in the Treatment of Skin Wounds in a Dog (Canis lupus familiaris)

Fernando Alzamora Filho 1, Manoel Luiz Ferreira 2, Marcus Vinícius Alves da Silva 2,
Anna Claudia Mombrini Silva Barbosa 3, Bruna Guedes Carvalho 4, Marcel Vasconcellos 5
,
José Marcus Raso Eulálio 2 & José Eduardo Ferreira Manso 2

ABSTRACT

Background: Laser photobiomodulation has been used in the treatment of various injuries and diseases. This promotes
modulation of the inflammatory process, edema reduction and devitalized tissue regeneration. The advantages of Anti-
microbial photodynamic therapy are its easy application and the absence of side effects. Other advantages are the cost
of the therapy, minimal damage to animal tissue, the broad spectrum of action, and efficiency against strains resistant
to antimicrobials. The aim of this study was to report the clinical and their resolution in a female dog with a traumatic,
infected wound treated with laser phototherapy as an alternative therapy method.
Case: A 3-year-old bitch Border Collie, weighing 18 kg, from the municipality of Ilhéus, Bahia, Brazil, was presented
for examination with a history of traumatic laceration of the left thoracic member. On the anamnesis, it was reported that
the patient presented with laceration of the left thoracic member. The wound was cleaned and an antibiotic [30 mg/kg of
12/12 h] and anti-inflammatory [0.1 mg/kg every 24 h were prescribed, both for 5 days]. Twenty-four h after the surgical
procedure, there was dehiscence of the sutures, with daily topical therapy based on zinc oxide for secondary intention
healing. Upon physical examination, the wound was found to be contaminated with swollen and erythematous edges, an
ulcerated area with devitalized tissue, serous exudate, and 8.8 cm2 of wounded area. Given the macroscopic characteristics
of the lesion, phototherapy was associated with conventional therapy until complete healing of the wound, with three
weekly applications at 48 h intervals. Initially, the wound was cleaned with saline solution at 0.9% and a single treatment
with aPDT was scheduled due to the high degree of contamination. The dosimetry parameters of irradiation were calcu-
lated according to the wounded area with a diode laser of 0.1 W of power, continuous emission, spot area of 0.028 cm2,
and energy of 9 J per application point. A gauze imbibed with 1 mL of methylene blue aqueous solution (300 μM), which
was the photosensitizer was applied to the lesion, with a pre-irradiation time of 5 min, after which it was irradiated with
red laser (RL) [λ = 660 nm] for 90 s per point, using the sweeping technique. The edge of the lesion was irradiated with
infrared laser (IRL) [λ = 808 nm], total energy of 5 J, using the technique of specific points and 1 J of energy/point. After
aPDT, low-intensity laser therapy (LILT) sessions were set up with RL and IRL, with energies of 0.5 J/point and 1 J/point,
respectively. The wound was cleaned daily, protected with a bandage, and clinically evaluated until complete regression.
Discussion: In the present case, methylene blue was used as a photosensitizer, but it is noteworthy that, apparently, each
microorganism responds differently to photosensitizers. Thus, the therapy becomes specific for each application, for example:
the type of photosensitizer, its concentration, pre-irradiation time, type of light used in photosensitization, wavelength,
energy, power, mode delivery of light. Thus, for the best result, the specific protocol in each application should be used
Low-intensity laser therapy is an easily executed technique with effective results. The use of PDT associated with photo-
biomodulation therapy enabled rapid healing of the cutaneous wound, in addition to an improvement in clinical signs and
pain caused by the lesion. The technique proved to be an efficient alternative in the treatment of wounds, whether used in
isolation or associated with conventional therapy.
Keywords: methylene blue, healing, wound, photobiomodulation, low-level laser.

DOI: 10.22456/1679-9216.116169
Received: 7 July 2021 Accepted: 24 September 2021 Published: 20 November 2021
1
Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz (UESC), Ilhéus, BA, Brazil. Centro de Cirurgia Experimental,
2

Departamento de Cirurgia, Faculdade de Medicina, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, Brazil. 3Secretaria Estadual de
Saúde do Espirito Santo (SESA-ES) Vitória¸ ES, Brazil. 4Universidade do Grande Rio (Unigranrio), Duque de Caxias, Rio de Janeiro, RJ. 5Faculdade
de Medicina, Centro Universitário Serra dos Órgãos (Unifeso), Teresópolis, RJ. CORRESPONDENCE: F. Alzamora Filho [[email protected]].
Universidade Estadual de Santa Cruz (UESC). Rodovia Jorge Amado, Km 16. CEP 45662-900 Ilhéus, BA, Brazil.

1
F. Alzamora Filho, M.L. Ferreira, M.V. A. Silva, et al. 2021. Antimicrobial Photodinamic Therapy Combined with Laser Photobiomodu-
lation in the Treatment of Skin Wounds in a Dog (Canis lupus familiaris). Acta Scientiae Veterinariae. 49(Suppl 1): 707.

INTRODUCTION and 8.8 cm2 of wounded area (Figure 1A). Given the
Rupture of the anatomical and physiological in- macroscopic characteristics of the lesion, phototherapy
tegrity of integumentary tissue can be caused by several was associated with conventional therapy until complete
factors [5]. Laser photobiomodulation (LPB) has been healing of the wound, with three weekly applications
used in the treatment of various injuries and diseases at 48 h intervals. Initially, the wound was cleaned with
[6,12]. For photophysical-chemical effects to occur, saline solution at 0.9% and a single treatment with aPDT
the endogenous chromophore photoacceptors need to was scheduled due to the high degree of contamination.
absorb the photons of irradiated light. This promotes The dosimetry parameters of irradiation were calculated
modulation of the inflammatory process, edema reduc- according to the wounded area with a diode laser of 0.1
tion and devitalized tissue regeneration [3,10]. Laser W of power, continuous emission, spot area of 0.028
therapy exhibit early epithelialization due to increased cm2, and energy of 9 J per application point. A gauze
neovascularization and fibroblast proliferation [11]. imbibed with 1 mL of methylene blue aqueous solution
Antimicrobial photodynamic therapy (aPDT) (300 μM), which was the photosensitizer (Figure 1B)
uses a non-toxic photosensitizer, specific light wave- was applied to the lesion, with a pre-irradiation time of 5
length, and molecular oxygen. Interaction of the triad min, after which it was irradiated with red laser (RL) [λ =
triggers a chemical reaction for the formation of oxygen 660 nm] for 90 s per point, using the sweeping technique
reactive species and singlet oxygen production, enabling (Figure 1C). The edge of the lesion was irradiated with
the destruction of microorganisms [4]. The advantages infrared laser (IRL) [λ = 808 nm], total energy of 5 J,
of aPDT are its easy application and the absence of side using the technique of specific points and 1 J of energy/
effects [2]. The short contact time of the photosensitizer point. After aPDT, low-intensity laser therapy (LILT)
with the bacteria and the simultaneous action on pro- sessions were set up with RL and IRL, with energies of
teins, lipids, and nucleic acids inhibit the expression of 0.5 J/point and 1 J/point, respectively. The wound was
protective factors such as stress proteins, which minimi- cleaned daily, protected with a bandage, and clinically
zes the possible appearance of acquired resistance [7]. evaluated until complete regression.
Other advantages are the cost of the therapy, minimal DISCUSSION
damage to animal tissue, a broad spectrum of action, and
efficiency against strains resistant to antimicrobials [12]. In this report, the use of aPDT was proposed as
The aim of this study was to report the clinical initial treatment as it causes the selective death of vari-
alterations and their resolution in a female dog with ous microorganisms [8]. Considering the macroscopic
a traumatic, contaminated wound treated with laser characteristics of the wound, with loss of tissue, swol-
phototherapy as an alternative therapy method. len edges, devitalization, and the presence of secretion,
in addition to failure of healing by first intention,
CASE
low-intensity laser therapy was also carried out. This
A 3-year-old bitch Border Collie, weighing 18 was due to its biomodulation effects, which promote
kg, from the municipality of Ilhéus, Bahia, Brazil, was cellular homeostasis and tissue regeneration [3].
presented for examination with a history of traumatic Forty-eight h after the first session, the lesioned
laceration of the left thoracic member. On the anamnesis, area showed reduced edema and erythema, granulation
it was reported that the patient presented with laceration tissue formation in the bed of the lesion, absence of
of the left thoracic member. The wound was cleaned exudate, and a reduction of 3.8 cm2. This improvement
and an antibiotic1 [Cefadroxila® - 30 mg/kg of 12/12 h] in the wound after one laser therapy session corroborated
and anti-inflammatory2 [Meloxicam® - 0.1 mg/kg every the literature, which described the production of oxygen
24 h] were prescribed, both for 5 days. Twenty-four h reactive species and singlet oxygen by aPDT that pro-
after the surgical procedure, there was dehiscence of moted the death of bacterial microorganisms. With the
the sutures, with daily topical therapy based on zinc addition of the effects of laser photobiomodulation, this
oxide3 (Alantol®) for secondary intention healing. favored the modulation of inflammation, and reduction
Upon physical examination, the wound was found to be of pain and edema [1]. They also promote a significant
contaminated with swollen and erythematous edges, an decrease in the lesioned area and the presence of gra-
ulcerated area with devitalized tissue, serous exudate, nulation tissue (Figure 2A).

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F. Alzamora Filho, M.L. Ferreira, M.V. A. Silva, et al. 2021. Antimicrobial Photodinamic Therapy Combined with Laser Photobiomodu-
lation in the Treatment of Skin Wounds in a Dog (Canis lupus familiaris). Acta Scientiae Veterinariae. 49(Suppl 1): 707.

The second session was with RL [λ = 660 nm] towards the center, covered by epithelial tissue, and hair
and IRL [λ = 808 nm], using energies of 0.5 J/point and growth, showing advanced morphofunctional restoration
1 J/point, respectively. In the third session, aPDT was of the involved tissues (Figure 2C).
carried out once again, due to the patient having conta- These results are caused by the photochemical
minated and injured the granulation tissue in the wound effect of laser therapy, which restores cellular homeo-
(Figure 2B). Two points were irradiated with 9 J of ener- stasis. Its effects can be observed in different stages
gy, RL on the ulcerated area and 5 stitches, with 1 J of of healing, with modulation of inflammation, neoan-
energy, and IRL on the edge of the lesion. The 4 following giogenesis, analgesia, accelerated collagen deposition,
sessions were with RL on the bloody area, at an energy of and endothelial cell proliferation [3,12].
0.5 J/point, and IRL on the edge, at an energy of 1 J/point, On the 14th day, the healing process was ad-
with a 48 h interval between sessions. A gradual increase vanced, with a 0.4 cm2 area of crust and the exuberant
in healing was observed, with the edge being displaced presence of hair in the shaved area (Figure 2D). The fi-

Figure 1. Contaminated wound with suture dehiscence. A- Devitalized wound, with swollen edges and serous exudate. B- Application of 0.01% methy-
lene blue (300 μM) on the contaminated wound. C- Irradiation with red laser (λ = 660 nm) for 90 s per point, on the lesioned region.

Figure 2. Healing evolution. A- Presence of granulation tissue and reduction of the wounded area 48 h after the phototherapy session. B- Day 5, photo-
dynamic therapy with 0.01% methylene blue photosensitizer. C- Day 10, wound with reduction of the lesioned area and cellular migration from the edge
of the wound, in the fifth session of phototherapy. D- Day 14, complete tissue repair, showing a scar with clear edges and advanced morphofunctional
reestablishment of the involved tissue, albeit with a slightly uneven central portion, in the seventh phototherapy session.

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F. Alzamora Filho, M.L. Ferreira, M.V. A. Silva, et al. 2021. Antimicrobial Photodinamic Therapy Combined with Laser Photobiomodu-
lation in the Treatment of Skin Wounds in a Dog (Canis lupus familiaris). Acta Scientiae Veterinariae. 49(Suppl 1): 707.

nal session of photobiomodulation was then conducted Low-intensity laser therapy is an easily execu-
[9] reported that laser therapy promotes the healing of ted technique with effective results. The use of PDT
chronic wounds in cases of reduced microcirculation. associated with photobiomodulation therapy enabled
In the present case, methylene blue was used as rapid healing of the cutaneous wound, in addition to
an improvement in clinical signs and pain caused by
a photosensitizer, but it is noteworthy that, apparently,
the lesion. The technique proved to be an efficient
each microorganism responds differently to photosen-
alternative in the treatment of wounds, whether used
sitizers. Thus, the therapy becomes specific for each in isolation or associated with conventional therapy.
application, for example: the type of photosensitizer, its
MANUFACTURERS
concentration, pre-irradiation time, type of light used in
photosensitization, wavelength, energy, power, mode 1
Vansil Saúde Animal. Descalvado, SP, Brazil.
2
Ourofino Saúde Animal. Cravinhos, SP, Brazil.
delivery of light. Thus, for the best result, the specific 3
Vetnil Indústria e Comércio de Produtos Veterinários Ltda.
protocol in each application should be used [1,2]. Louveira, SP, Brazil.

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