Key Parameters in Phototherapy With Gold Nanorods Using Continuous Near Infrared

Research Article

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Key Parameters in Phototherapy with Gold Nanorods Using

Continuous Near Infrared Radiation
Sara C. Freitas, João H. Belo, Andreia Granja, Miguel Canhota, Ana S. Silva,
Salette Reis, Helder Crespo, João P. Araújo, and Célia T. Sousa*
1. Introduction
As nanoparticle formulations move toward human clinical trials in photo-
thermal cancer therapy (PTT), the influence of individual key parameters In recent years, hyperthermia emerged
on the heating efficacy must be thoroughly assessed. This work reports a as a promising treatment approach in
oncology, consisting in raising the tem-
systematic study on the heating performance of gold nanorods during
perature of cancer cells to 40–45 °C to
exposure to near-infrared radiation, evaluating the influence of nanorods reach apoptosis, i.e., programmed cell
concentration, total volume, laser output power, and spot area. Interestingly, death.[1,2] One way to reach local and
the lowest concentration tested (24 µg mL−1) shows the most promising results controlled hyperthermia is via function-
with a SAR (Specific Absorption Rate) value of 24.6 kW gAu−1 for the highest alizable nanostructures that are activated
laser power (0.8 W), spot area (0.4 cm2), volume (1 mL). The laser output by external stimuli, such as electro-
magnetic radiation or magnetic fields.
power and concentration proved to be the key parameters in global heating of
In this context, photothermal therapy
the sample. The cuvette’s optical path length also proves to be an important (PTT) is a minimally invasive local treat-
parameter given that there is a threshold concentration value beyond which ment modality whose goal is to convert
no significant improvement will be observed, and the higher gold mass will electromagnetic radiation into heat by
play a detrimental role suppressing SAR values. It is experimentally demon- stimulation of photoabsorbing agents
that are administrated to the body intra-
strated that the multi-parameter exploration can lead to a finer control of the
venously or intratumorally.[1,2] Laser
performance in PTT, opening a pathway for efficient heating of low nanopar- light in near-infrared (NIR) region is the
ticle concentrations. energy source typically used in PTT due
to its enhanced tissue penetration capa-
bility with lower absorption in biological
tissues, that limits the heating of healthy surrounding tis-
S. C. Freitas, J. H. Belo, M. Canhota, A. S. Silva, H. Crespo, sues.[2] Among several nanomaterials, gold nanorods (AuNRs)
J. P. Araújo, C. T. Sousa have been extensively explored as photothermal agents due to
IFIMUP their biocompatibility and ability to generate heat due to the
Departamento de Física e Astronomia da Faculdade
de Ciências da Universidade do Porto absorption of electromagnetic radiation. The AuNRs heat gen-
Rua do Campo Alegre, 687, 4169-007 Porto, Portugal eration phenomenon, induced by the laser irradiation, can be
E-mail: celiasousa@fc.up.pt explained by a particular feature of metallic nanoparticles—
A. Granja, S. Reis surface plasmon resonance (SPR).[1–3] SPR is caused by the
LAQV presence of conduction electrons that oscillate on the metallic
REQUIMTE
nanoparticle surface. If the incident light wavelength is in
Faculdade de Farmácia da Universidade do Porto
R. Jorge de Viterbo Ferreira 228, 4050-313 Porto, Portugal resonance with the oscillating electrons, a high-energy state
C. T. Sousa is reached, leading to a sequential relaxation in the form of
Departamento de Física Aplicada heat. In this case, the light absorption results in optimal heat
Universidad Autónoma de Madrid, Avda. Francisco Tomás y Valiente 7 generation that ultimately dissipates from the particle to the
Campus de Cantoblanco, 28049 Madrid, Spain surrounding media.[1,2]
E-mail: celia.tsousa@uam.es
Phototherapy mediated by gold nanoparticles has already
shown promising results in vitro and in vivo (with animal
The ORCID identification number(s) for the author(s) of this article
can be found under https://doi.org/10.1002/admi.202202214. models) for different types of cancer. Manivasagan et al.
demonstrated that gold nanorods ([AuNR] = 25 µg mL−1),
© 2023 The Authors. Advanced Materials Interfaces published by duly modified for biocompatibility purposes and combined
Wiley-VCH GmbH. This is an open access article under the terms of the with laser irradiation (2 W cm−2, 5 min) induced significant
Creative Commons Attribution License, which permits use, distribution apoptosis (63.3%) when compared with three control groups
and reproduction in any medium, provided the original work is properly
cited. (0.38%, 1.74%, and 12.01%) on breast cancer cells (MDA-MB-
231).[4] The same study also showed that 808 nm NIR laser
DOI: 10.1002/admi.202202214 irradiation on tumor-bearing mice filled with these AuNRs
Adv. Mater. Interfaces 2023, 10, 2202214 2202214 (1 of 8) © 2023 The Authors. Advanced Materials Interfaces published by Wiley-VCH GmbH
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lead to a complete tumor removal with no regrowth observed 2. Results and Discussion
over the therapeutic period of 20 days.[4] In parallel, Sangnier
et al. studied the photothermal effect of four gold nanoparti- 2.1. Gold Nanorod Characterization
cles morphologies and the resulting induced cancer cell death
within extra or intracellular localizations (internalized before Figure 1a,b presents, respectively, a scanning electron micros-
PTT) on human prostate cancer (PC3) cells. For cancer cells copy (SEM, FEI Quanta 400FEG high resolution (HR)) image
exposed to 808 nm NIR irradiation (0.3 W cm−2, 10 min) of the gold nanorods before and after laser irradiation. These
with gold nanorods ([AuNR] = 98.5 µg mL−1) the number of data demonstrates that laser irradiation does not affect the mor-
cancerous viable cells decreases to 38% in the extracellular phology of the AuNRs, and that their shape is maintained intact
situation and to 11% in the intracellular situation, this way after successive irradiations. Before irradiation the AuNRs
highlighting the effectiveness of PTT treatment.[5] Taking present an average length and diameter of 51.7 ± 9.3 nm and
advantage of the surface modification capability of gold nanorods 12.2 ± 3.4 nm, respectively. After irradiation the dimensions are
with specific antibodies that promote cancer-cell targeting, similar, 55.4 ± 10.5 nm and 13.7 ± 3.2 nm (Figure 1c,d). These
Zhang et al. reported the autophagic effect (alternative cell nanostructures exhibit plasmon resonance peaks, as observed
death mechanism to apoptosis) of gold nanorods functional- on the absorbance spectra of Figure 1e, with maximum around
ized with anti-epidermal growth factor receptor (EGFR) com- 497 and 801 nm. These peaks are related, respectively, to the
bined with NIR-PTT (100 nm, 1.5 W cm−2, 3 min) in a mouse transversal and longitudinally localized surface plasmon reso-
xenograft model (MDA-MB-231 cell lines). The results showed nances (LSPR) of AuNRs with aspect ratio close to 4, this way
that the combined treatment of NIR-PTT with anti-EGFR- covering the NIR window.[2,3,10]
AuNRs caused a significant reduction in the tumor volume
(down to 22.67 ± 8.60% mm3) when compared to the control
group (average tumor size of 150–200 mm3).[6] Maltzahn et al. 2.2. Role of Laser Output Power and the Effect of Concentration
showed that polyethylene glycol (PEG) coated AuNRs intrave-
nous injected into tumor-bearing mice (20 mg Au kg−1) could The results presented in this sub-section are related to the
accumulate in tumor (≈7% ID g−1 at 72 h post injection) and be influence of the laser output power and the gold nanorods con-
rapidly heated to over 70 °C by NIR laser irradiation (2 W cm−2, centration. Here, the heating curves were obtained in a fixed
5 min). In their work, within 10 days all the irradiated PEG- solution volume (1 mL) and laser spot area (0.4 cm2) for the con-
AuNR-targeted tumors were completely eradicated with no evi- centrations 24, 75, and 200 µg mL−1 in a 1 cm optical path (OP)
dence of tumor regrowth.[3,7] All these mentioned reports eval- cuvette. The laser output power was varied in the 0.2–0.8 W (or
uated the effectiveness of PTT through cell viability studies, power density 0.5–2 W cm−2) range for each concentration. For
but the figure of merit SAR (Specific Absorption Rate) is also all cases, the heating curves were also obtained in an only-water
often employed to compare the nanoparticle heating efficiency solution of the same volume to infer about the effective contri-
in a given environment.[1,5,8,9] In this context, Espinosa et al. bution of the gold nanorods in the sample heating. This contri-
reported an intensive study that evaluated the influence of dif- bution of the temperature variation on water-only solution was
ferent shaped gold nanostructures with regard to peak absorp- not subtracted on the subsequent heating curves obtained with
tion position and heat generation efficiency.[1] Among all the the AuNRs diluted, but its results are plotted for comparison.
studied nanostructures AuNRs showed the larger temperature All the values related to maximum temperature variation (fitted
raise (ΔT ≈ 20 °C). For a [AuNR] = 150 µg mL−1 in an aqueous parameters) and SAR (W gAu−1) calculated from the data pre-
dispersion, SAR values reached 3 and 10 kW gAu−1, at laser sented in Figure 2 are summarized in Table S1 (Supporting
power densities of 0.3 or 1 W cm−2, respectively.[1] In 2020, Information). The photo-induced heating curves are presented
Sangnier et al. reported a similar study and in apparently in Figure 2a where it is noticeable that after a few minutes
similar conditions (aqueous solution, 808 nm NIR radiation, under radiation exposure a plateau temperature is reached.
[AuNRs] = 150 µgAu mL−1 and 0.3 W cm−2) the estimated SAR At this stage the heating and the losses to surroundings are
values were ≈6 kW gAu−1, which is twice the value previously already counterbalanced, keeping the temperature of the solu-
mentioned.[5] Given the discrepancies, what are the parame- tion stable. Independently of the nanorods concentration and
ters that truly influence the SAR? How each individual experi- the laser power, the temperature plateau is reached few min-
mental parameter affects the global heating of a sample? This utes (around 400 s) after laser irradiation starts.
work aims to fill some of these knowledge gaps, reporting a Observing Figure 2a, one can see that the laser output power
systematic study on the heating performance of such nano- parameter has a clear influence on the temperature variation of
structures during exposure to NIR radiation. Throughout the the solution. A maximum value of ∆T = 25.9 °C was achieved for
experiments several parameters were considered, including 200 µg mL−1 (excluding the initial 5 s[11,12]) and decreasing the con-
the laser output power (W) or power density (W cm−2), spot centration to 24 µg mL−1, a ∆T = 20.4 °C is reached. It is seen that
area, AuNRs concentration and the volume of the aqueous the greatest temperature variation is associated with the largest
solution in which the latter were suspended. Knowing how laser output power employed. This can be explained based on the
each one of these parameters influences the global heating of heat contribution of a large number of individual gold nanorods,
a sample, one can optimize the AuNRs heating performance where each generates a certain amount of heat (Q, W) under laser
which will be translated into more efficient photothermal irradiation. The Q value can be obtained as the product of absorp-
therapy protocols. tion cross-section area (Cabs, m2) and laser fluence (I, W m−2):[8]
Figure 1. Scanning electron microscopy (SEM) images of the gold nanorods a) before laser irradiation. b) after laser irradiation. c) Gold Nanorod
length distribution before (51.7 ± 9.3 nm) and after (55.4 ± 10.5 nm) irradiation. d) Gold Nanorod diameter distribution before (12.2 ± 3.4 nm) and
after (13.7 ± 3.2 nm) irradiation. e) Absorbance spectra of AuNRs not irradiated (red) and irradiated (black curve) with the highest laser power (0.8 W),
obtained at concentration [Au] = 75 µg mL−1.
Q = Cabs·I (1) increase with the laser power, but the solution of 24 µg mL−1
clearly stands out reaching 2.45 × 105 W gAu−1 (Figure 2f). This
Therefore, for each concentration with a given cross sec- means that larger concentrations do not necessarily reflect
tion area, the amount of heat generated is directly proportional greater heating efficiency. In fact, the results suggest precisely
to the laser power. This is clearly reflected in the obtained the opposite, revealing that the solution of lower concentration
results, where for each concentration a linear increase of max- presents the best heating performance.
imum temperature variation with the laser power can be seen
(Figure 2d). However, the heating rate, dT/dt, decreases when
increasing the concentration (Figure 2e), which is associated to 2.2.1. Role of the Cuvette’s Optical Path
a decrease in the Cabs, that corresponds to the effective area that
absorbs the light energy impinging upon the nanorods and is An explanation for the outcome results presented in the pre-
strongly dependent on the concentration under resonant con- vious section could be related to the cuvette´s optical path
ditions. It is also possible to verify that, regardless of the used and laser penetration depth. Just to recall, the results obtained
concentration, the maximum temperature variation does not in a 1 cm OP cuvette showed a significant change in ΔT (°C)
undergo major changes (Figure 2e). By increasing the concen- between the first two concentrations (24 and 75 µg mL−1) that
tration from 24 to 200 µg mL−1 only a small increase in the ∆T of no longer holds for the third (200 µg mL−1). This result suggests
1.4 and 5.5 °C was achieved, respectively, for the lowest (0.2 W) that for the highest concentration, the laser beam energy is not
and highest (0.8 W) laser powers. The same cannot be said crossing through the sample and consequently the AuNRs pre-
regarding its heating efficiency (detailed later in Section 4.1), sent in the solution are not being evenly illuminated. In such
evaluated as dissipated power per unit of mass of mate- model, there is a threshold in AuNRs concentration value,
rial (W gAu−1). In this regard, one can see that all SAR values above which the solution becomes completely opaque to NIR
Figure 2. Role of AuNR concentration and laser output power. a) Typical photo-induced heating curves of AuNRs upon continuous NIR radiation
(0.2–0.8 W range) for the concentrations 24 µg mL−1. b) Same as previous but for [AuNRs] = 75 µg mL−1. c) Same as previous but for [AuNRs] = 200 µg mL−1.
d) Maximum temperature variation as a function of laser output power for each concentration (24, 75, and 200 µg mL−1) and water-only solution.
e) Maximum temperature variation as a function of AuNR concentration upon continuous NIR radiation (0.2–0.8 W range). f) SAR (W gAu−1) as a
function of AuNR concentration for each laser output power (0.2–0.8 W range).
beam, AuNRs become unevenly illuminated and the maximum Contrarily to the previously obtained results in the 1 cm OP
ΔT (°C) values observed saturate. This translates into a decrease cuvette (where the maximum temperature variation, ΔT, did
in the Cabs and subsequently, into a lower amount of heat gen- not undergo major changes regardless of the used concentra-
eration. To verify if effectively higher concentrations (such as tion), here ΔT is clearly seen to increase with AuNRs concen-
200 µg mL−1) may be sufficient to make the solution “opaque” tration (Figure 3a). For the highest laser power, a ΔT of 18.2
to NIR radiation we repeated the experiment with a narrower and 28.6 °C was reached for the concentrations of 75 and
cuvette with 1 mm OP and 0.5 mL volume. Results are pre- 200 µg mL−1, respectively (in the 1 mm OP cuvette), in con-
sented in Figure 3. trast with a much more subtle ΔT difference between 24.6 and
Figure 3. a) Maximum temperature variation as a function of laser output power for two AuNR concentrations (75 and 200 µg mL−1) and water-only
solution b) SAR (W gAu−1) as a function of laser output power (0.2–0.8 W range) for two AuNR concentrations (75 and 200 µg mL−1). Measurements
were acquired on 1 mm OP cuvette with 0.5 mL solution volume.
25.9 °C that was reached for the concentrations of 75 and observed in ΔT values and simultaneously the higher Au mass
200 µg mL−1, respectively (in the 1 cm OP cuvette). This new will play a detrimental role on suppressing SAR values, as can
observed trend supports the hypothesis that the laser unevenly be seen in Figure 3b.
illuminates all AuNRs in the solution as it must be above the
threshold concentration value for a 1 cm OP cuvette.
In addition, we also performed laser power measurements 2.3. Role of Laser Spot Area
in the positions immediately before and after the cuvette’s to
find out how much power was lost in the path of laser direc- The following results were obtained with a focusing lens
tion for every irradiation experiment. The results are organized included in the set-up to shape the incident beam spot size
in Tables 6 and 7 (Supporting Information) and presented in area. Placing this lens at different distances along the beam
Figure S2 of (Supporting Information). Remarkably, with the path, allowed to study the effect of the laser spot area, while
smallest concentration (24 µg mL−1) there is already 61% of the laser power density was kept fixed. Here, the results
power loss across the 1 cm cuvette. Here, even the 75 µg mL−1 were acquired for the highest concentration (200 µg mL−1) in
is already approaching to the saturation threshold, allowing three laser spot areas: i) 0.4, 0.091, and 0.054 cm2, in a 0.5 mL
only 2.23% of incident power to reach the back of the cuvette. solution and with a 1 mm OP cuvette. The laser power was
With the highest concentration nearly all the laser power is adjusted in each case so that the laser power density remains
“retained” inside the cuvette and less than 1% reaches its back, equal in all experiments, in the range of 0.5-2 W cm−2.
confirming once again that the solution is sufficiently concen- Observing Figure 4 one can see that ΔT, and consequently
trated to become opaque to NIR radiation. The same kind of SAR, increase with the size of the laser spot area, reaching a
behavior in observed for the 1 mm OP cuvette, however, in this maximum 7.09, 9.24, and 28.6 °C for the highest power density
case the solution is not yet saturated and there is still some (2 W cm−2) in the increasing areas of 0.054, 0.091, and 0.4 cm2,
power that completely crosses the entire cuvette (about 12.1% respectively. Here, since we kept the power density fixed, that
for the 200 ug mL−1 concentration). These results demonstrate, corresponds to the number of photons per unit area, what is
that for every optical path length there is a threshold concen- changing is the number of nanorods that are “active” under the
tration value beyond which no significant improvement will be laser irradiation and contributing to the heating. As the amount
Figure 4. Role of the laser spot area a) Photo-induced heating curves of AuNRs upon continuous NIR radiation (0.5–0.2 W cm−2 range) for the concen-
tration 200 µg mL−1 in a laser spot area of 0.4 cm2. b) Same as previous but for spot area 0.091 cm2. c) Same as previous but for spot area 0.054 cm2,
d) SAR (W gAu−1) as a function of laser power density for each laser spot area (0.4, 0.091, and 0.054 cm2). e) Maximum temperature variation as a
function of laser power density for each laser spot area (0.4, 0.091, and 0.054 cm2).
of generated heat becomes strongly increased in complexes variations were achieved for larger volumes, reaching the final
composed of several Au NPs, if a larger number of nanorods temperatures of 30.3 and 33.4 °C for the increasing volumes of
are involved, due to a larger spot area, more heat is being 0.3 and 0.5 mL, respectively. On one hand, this was an expected
generated.[13,14] result since larger volumes contain a higher amount of gold
nanorods (for a constant concentration) to absorb the NIR radi-
ation and release it in the form of heat. On the other hand, it
2.4. The Effect of Sample Volume is also true that the greater the volume, the greater the amount
of heat that must be supplied to the solution to increase its
The last parameter under evaluation was the volume of the temperature until stabilization. This can be observed on
aqueous solution in which the AuNRs were suspended. Here, the saturation temperature plateau, that is reached sooner
a given area of the laser spot was fixed (0.03 cm2) and the for smaller volumes (Figure 5b). In other words, the temperature
volume of the solution was varied within. The acquired data, stabilization, arising from the heat exchanges with the environ-
once more, included the heating curves for the three concentra- ment (liquid solution–air interface), happens faster when the
tions (24, 75, and 200 µg mL−1) on the 0.2–0.8 W power range, sample has less material. In fact, by directly comparing the τ
for three different solution volumes (0.3, 0.5, and 1 mL). It is fitted values one can see that for all concentrations, the latter
worth to mention that different cuvettes were used throughout is inferior for smaller volumes (0.3 mL). Here, τ is a character-
the experiments for set-up convenience. For the measure- istic heating time that depends on sample properties (described
ments performed on 1 mL solutions, a regular quartz cuvette later in Experimental Section). In Figure 5d, the τ values for
(V = 3500 µL) was used but a smaller one (V = 1400 µL) was the 0.5 and 1 mL volumes present very similar results, regard-
required for the remaining evaluated volumes (Figure 5a). less of the concentration. At this point, it is worth to recall
Also, it was verified that, in both cuvettes, the entire area of the that the 1 mL volume measurements were obtained in a reg-
laser spot was always within the cuvette´s surface area to avoid ular cuvette, whereas the remaining volumes were obtained
losses of intensity in its walls. with a small volume one, which has a narrower liquid sup-
Figure 5b demonstrates the heating curves acquired for the port. Considering the heat exchanges with the surroundings
lowest concentration (24 µg mL−1), power (0.2 W), and spot arising from losses by conduction through the cuvette walls,
area (0.03 cm2). Here it is noticeable that highest temperature the heat transfer (Qcond) of the sample is proportional to the
Figure 5. Role of sample volume. a) Schematic representation of the used cuvettes; b) Typical photo-induced heating curves of AuNRs upon con-
tinuous NIR radiation (0.2 W) for the sample volumes 0.3 and 0.5 mL in the reduced spot area (0.03 cm2); c) SAR (W gAu−1) as a function of laser
output power for each concentration (24, 75, and 200 µg mL−1); d) τ fitted values as a function of laser output power (0.2–0.8 W range) for the smallest
concentration (24 µg mL−1) in each solution volume (0.3, 0.5, and 1 mL); e) Same as previous but for [AuNRs] = 75 µg mL−1; f) Same as previous but
for [AuNRs] = 200 µg mL−1.
temperature difference across the layer and the heat transfer parameters, such as the AuNR concentration, laser power, laser
area, and inversely proportional to the thickness of the wall. spot area, and solution volume, on the gold nanorods heating
This is described by Fourier’s law of heat conduction, given by: efficiency. Throughout the experiments, three different AuNR
concentrations (24, 75, and 200 µg mL−1) were irradiated with a
∆T laser power density in the 0.5–2 W cm−2 range, on three laser
Q cond = k·A · (2)
∆x spot areas (0.4, 0.091, and 0.054 cm2). Afterward, the influence
where k is the thermal conductivity of the material (quartz, of the volume of solution (0.3, 0.5, and 1 mL) was also studied.
3 W m−1 K -1), A is the surface area, ΔT is the temperature dif- Among the evaluated conditions, the concentration and laser
ference across the wall and Δx is the thickness of the layer.[15] output power proved to be key parameters in phototherapy with
This implies that the heating of a sample does not only depend gold nanorods using continuous radiation. In all cases, the SAR
on the amount of material (represented here in volume of solu- values increase with laser output power reaching a maximum
tion under irradiation) but also on the surface area of the cuvette value of 2.46 × 104 W gAu−1 for the highest power (0.8 W) and
through which the heat exchanges take place. For comparison smallest concentration (1 mL, 0.4 cm2 spot). A major consistent
purposes and considering the four side walls of the cuvettes result verified under all evaluated conditions, was the fact that
in contact with the solution, the estimated area available for the lowest concentrations (24 µg mL−1) present higher SAR
heat exchanges is, respectively, 4, 3.64 and 2.52 cm2 for the 1, values, thus being more efficient for this application. In addi-
0.5, and 0.3 mL volumes. Here, the decrease in surface area, tion to this, it was also verified that for every cuvette’s optical
and in solution volume, is accompanied by a decrease in the path length there is a threshold concentration value beyond
τ values. The first two volumes present average τ values of 167.1 s which no significant improvement will be observed in ΔT
and 127.4 s for the volume of 1 mL and 0.5 mL, respectively, on values and simultaneously the higher Au mass will play a detri-
the 24 µg mL−1 concentration. The smaller volume (0.3 mL), mental role on suppressing SAR values. Above this concentra-
with a lower sectional area, presented an average τ = 111.3 s tion the solution becomes opaque to NIR radiation.
(Figure 5d). The reproducibility of the measurements was veri- The solution volume, on the other hand, highlight the
fied with several experiment repetitions, which resulted in an dependence of the acquired results on set-up variations or data
invariability of the τ. This means that all experiments were car- acquisition process. This means that the nanoparticle heating
ried out in the same thermal bath composed by the cuvette and efficiency is highly sensitive not only to the experimental condi-
the room where the measurements were carried out. Despite tions but also on the systematic errors introduced during the
the changes on temperature variation and heating rate of the measurements. A proper calibration of the thermocouple and
sample, the SAR values do not undergo so evident variations its fixed position during irradiation are crucial factors for accu-
(Figure 5c). As mentioned, the results suggest that the superior rate data gathering. Even the SAR calculation method itself can
heating rate is obtained for smaller volumes (B fitted parameter, lead to large variations in its value, depending on the initial
B = 1/τ), but the latter counterbalances with the superior tem- assumptions considered by researchers. The results obtained
perature variations (A fitted parameter) of larger volumes so in this work can be used as a starting point for future tests in
that, in the final calculation of the SAR (proportional to A* B) more complex models, as in in vivo tests, as our multi-param-
the values remain close to each other. For the lowest concentra- eter exploration can lead to a finer control of the performance
tion (24 µg mL−1) and power (0.2 W), the calculated SAR values in PTT, opening a pathway for efficient heating of low nanopar-
were 4.20 × 103 W, 5.87 × 103 W, and 4.84 × 103 W gAu−1 for the ticle concentrations.
increasing volumes of 0.3, 0.5, and 1 mL. Based on the results,
the volume of the solution did not turn out to be a key para-
meter for the nanoparticle heating efficiency but underlined very 4. Experimental Section
clearly its dependence and susceptibility on the set-up and irradi-
ation conditions. The temperature variation with the laser power Materials and Methods: The measurements here reported were
in water solution was also measured for the two studied volumes performed using an infrared continuous laser (MDL-III-808/1-2500 mW)
operating at 808 nm. The laser was aligned in a way that its beam
(0.5 and 1 mL) in the same 1 cm OP cuvette (Figure 3 of the laterally crosses the walls of a glass cuvette filled with the AuNRs
Supporting Information). It is observed that, for both volumes, solution. The heating of the sample was measured with a type-K
ΔT (°C) linearly increases with power and that, for the same thermocouple and continuously confirmed with a FLIR i7 Infrared
power, the larger volume presents larger ΔT (°C). For the highest Thermal camera. Temperature measurements made by the thermal
power (0.8 W), the maximum registered ΔT was 8.62 and 17.4 °C camera showed the same values with the solution irradiated with
for 0.5 and 1 mL, respectively. Therefore, contrary to the con- or without the thermocouple, confirming that this as no influence
on the measurements. The laser power measurements were
centration effect, which is a parameter highly dependent on the performed with a Thorlabs Power Meter Console (PM100A). The gold
plasmonic effect of the material, the effect of volume is purely a nanorods (10 × 41 nm, SPR = 808 nm) with reference 035E12-10-808-
thermodynamic problem that depends on the environment sur- NPO-CHL-200-0.25 were purchased from Nanopartz (Nanopartz,
rounding the sample under heating and irradiation conditions. Loveland, CO, USA). The AuNRs aspect ratio (AR), given by the
proportion between their dimensions (length/width), is ≈4 having
therefore the ideal shape to be excited by the 808 nm laser.[3,10,16]
Throughout the experiments three different AuNR concentrations
3. Conclusions (24, 75, and 200 µg mL−1) were irradiated with a laser output power in
the 0.2–0.8 W range. Beyond concentration and laser output power,
The photo-induced heating tests performed in this system- the influence of the sample volume (0.3, 0.5, and 1 mL) and laser spot
atic work demonstrated the influence of several experimental area (0.40, 0.091, and 0.054 cm2) were also evaluated. Considering the
original and largest laser spot area (0.40 cm2), the applied fluences SFRH-BPD-87430/2012. AG acknowledges funding from the HEALTH-
lies under the generally laser powers densities employed on preclinical UNORTE I&D&I project (NORTE-01-0145-FEDER-000039) co-financed
PTT studies.[1,3,4,7] In this case, the used power densities were on the by European Regional Development Fund (ERDF), through the NORTE
0.5–2 W cm−2 range. 2020 (Programa Operacional Regional do Norte 2014/2020).
The acquired data included temperature stabilization (200 s), the
heating of the sample upon irradiation until temperature saturation and
the respective cooling once the laser was turned off. The samples were Conflict of Interest
under laser irradiation for ≈16.7 min (≈1000 s). The obtained temperature
The authors declare no conflict of interest.
versus time curves were used to calculate the SAR (Specific Absorption
Rate) values by one of the most popular methods found in the literature:
the Box–Lucas method (BLM).[11,12,17,18] This method, employed as a
figure-of-merit to quantify and compare NP heating efficiency, relies on
Data Availability Statement
data fitting and analysis and supposes a non-adiabatic set-up where The data that support the findings of this study are available in the
convective heat losses were taken under consideration.[12] Therefore, Supporting Information of this article.
the BLM is an appropriate model for the sample temperature-time
dependence that includes heat exchanges between the sample and
the surroundings.[18] The Box–Lucas equation for the non-adiabatic Keywords
temperature increase can be expressed as:
continuous laser, gold nanorods, oncology, phototherapy
T ( t ) = A (1− e −Bt ) + C (3)
Received: October 11, 2022
where the parameters A and B are, respectively, related with the final Revised: February 17, 2023
temperature (i.e., A = T(t = ∞)) and the heating rate of a given sample Published online: March 25, 2023
(B = 1⁄τ). Here, τ is a characteristic heating time that depends on sample
properties. The product of these parameters A*B represents the heat
rate and can be used to calculate SAR by the following formulae:
[1] A. Espinosa, J. Kolosnjaj-Tabi, A. Abou-Hassan, A. Plan Sangnier,

·c · 
mw dT
SARBLM = (4)
mAu w  dt  max A. Curcio, A. K. A. Silva, R. Di Corato, S. Neveu, T. Pellegrino,
L. M. Liz-Marzán, C. Wilhelm, Adv. Funct. Mater. 2018, 28, 1803660.
 dT  = A·B [2] A. Granja, M. Pinheiro, C. T. Sousa, S. Reis, Biochem. Pharmacol.
 dt  max (5)
2021, 190, 114639.
[3] L. Cheng, C. Wang, L. Feng, K. Yang, Z. Liu, Chem. Rev. 2014, 114,
mw (6) 10869.
SARBLM = ·c · A·B
mAu w [4] P. Manivasagan, S. Bharathiraja, S. Moorthy, S. Mondal,
T. P. Nguyen, H. Kim, T. T. V. Phan, K. Lee, J. Oh, Polymers 2018, 10,
where cw is the specific heat capacity of water (4.186 J g−1 °C−1) and
232.
mAu is the total mass of gold in the sample. The heating rate of the
[5] A. Plan Sangnier, A. Van De Walle, R. Aufaure, M. Fradet, L. Motte,
sample was obtained by fitting the curve during sample cool down
E. Guénin, Y. Lalatonne, C. Wilhelm, Adv. Biosyst. 2020, 4, 1900284.
and the final temperature was obtained fitting the heating curve during
laser irradiation.[11,12,16,18] For accurate SAR calculations, the timeframe [6] M. Zhang, J. Photochem. Photobiol., B 170, 58.
selected for the fit should exclude the initial 5 s. In Box–Lucas’ fitting, [7] G. von Maltzahn, J.-H. Park, A. Agrawal, N. K. Bandaru, S. K. Das,
including this initial non-linear heating response results in a decrease in M. J. Sailor, S. N. Bhatia, Cancer Res. 2009, 69, 3892.
the exponential factor that fits the curvature of the temperature change [8] Z. Qin, J. C. Bischof, Chem. Soc. Rev. 2012, 41, 1191.
as a function of time and could lead to inaccurate SAR values.[11,12] [9] C. Yao, L. Zhang, J. Wang, Y. He, J. Xin, S. Wang, H. Xu, Z. Zhang,
J. Nanomater. 2016, 2016, 5497136.
[10] D. Jaque, L. Martínez Maestro, B. del Rosal, P. Haro-Gonzalez,
A. Benayas, J. L. Plaza, E. Martín Rodríguez, J. García Solé,
Supporting Information Nanoscale 2014, 6, 9494.
[11] R. Wildeboer, P. Southern, Q. Pankhurst, J. Phys. D: Appl. Phys. 2014,
Supporting Information is available from the Wiley Online Library or
from the author. 47, 495003.
[12] H. L. Ring, A. Sharma, R. Ivkov, J. C. Bischof, Int. J. Hyperthermia
2020, 37, 100.
[13] H. H. Richardson, M. T. Carlson, P. J. Tandler, P. Hernandez,
Acknowledgements A. O. Govorov, Nano Lett. 2009, 9, 1139.
[14] A. O. Govorov, W. Zhang, T. Skeini, H. Richardson, J. Lee,
This work was financially supported by Funded by project N. A. Kotov, Nanoscale Res. Lett. 2006, 1, 84.
UIDB/04968/2020 from FCT. The authors also acknowledge to the
[15] S. Kakaç, Y. Yener, C. P. Naveira-Cotta Heat Conduction, 5th ed., CRC
European Union’s Horizon 2020 research and innovation program
Press, an imprint of Taylor and Francis, Boca Raton 2018.
under the Marie Sklodowska-Curie Grant Agreement No. 734801, CERN/
[16] S. K. Sharma, N. Shrivastava, F. Rossi, L. D. Tung, N. T. K. Thanh,
FISTEC/0003/2019, PTDC/EME-TED/3099/2020, 203269-ExtreMed-
NORTE-01-0247-FEDER-045932, UIDP/04968/2020 and to the Nano Today 2019, 29, 100795.
Network of Extreme Conditions Laboratories (NECL) under the project [17] O. L. Lanier, O. I. Korotych, A. G. Monsalve, D. Wable, S. Savliwala,
NORTE-01- 0145-FEDER-022096. CTS acknowledges to the program N. W. F. Grooms, C. Nacea, O. R. Tuitt, J. Dobson, Int. J. Hyper-
Atraccion de Talento (CAM), ref. 2020-T1/IND-19889. J. H. Belo thermia 2019, 36, 686.
acknowledges EEA grants for the project FBR_OC1_85 and PARSUK-FCT [18] B. Sanz, M. P. Calatayud, N. Cassinelli, M. R. Ibarra, G. F. Goya, Eur.
for the project SMARTX and for his contract DL57/2016 reference J. Inorg. Chem. 2015, 2015, 4524.

Key Parameters in Phototherapy With Gold Nanorods Using Continuous Near Infrared

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Key Parameters in Phototherapy With Gold Nanorods Using Continuous Near Infrared

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Key Parameters in Phototherapy with Gold Nanorods Using

[1] A. Espinosa, J. Kolosnjaj-Tabi, A. Abou-Hassan, A. Plan Sangnier,

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