Pflügers Archiv - European Journal of Physiology (2021) 473:785–792

https://doi.org/10.1007/s00424-020-02428-8

ORIGINAL ARTICLE

tDCS effects on brain network properties during physiological aging

Fabrizio Vecchio 1 & Francesca Miraglia 1 & Claudia Rodella 2 & Francesca Alù 1 & Carlo Miniussi 2,3 & Paolo Maria Rossini 1 &
Maria Concetta Pellicciari 2

Received: 11 March 2020 / Revised: 5 June 2020 / Accepted: 26 June 2020 / Published online: 4 July 2020
# Springer-Verlag GmbH Germany, part of Springer Nature 2020

Abstract
Brain neural networks undergo relevant changes during physiological aging, which affect cognitive and behavioral functions.
Currently, non-invasive brain stimulation techniques, such as transcranial direct current stimulation (tDCS), are proposed as tools
able to modulate cognitive functions in brain aging, acting on networks properties and connectivity. Segregation and integration
measures are used and evaluated by means of local clustering (segregation) and path length (integration). Moreover, to assess the
balancing between them, the Small World (SW) parameter is employed, evaluating functional coupling in normal brain aging and
in pathological conditions including neurodegeneration. The aim of this study was to systematically investigate the tDCS-
induced effects on brain network proprieties in physiological aging. In order to reach this aim, cortical activity was acquired
from healthy young and elderly subjects by means of EEG recorded before, during, and after anodal, cathodal, and sham tDCS
sessions. Specifically, the aim to exploring tDCS polarity-dependent changes in the age-dependent network dynamics was based
on a network graph theory application on two groups divided in young and elderly subjects. Eighteen healthy young (9 females;
mean age = 24.7, SD = 3.2) and fifteen elderly subjects (9 females; mean = 70.1, SD = 5.1) were enrolled. Each participant
received anodal, cathodal, or sham tDCS over the left prefrontal cortex (PFC) in three separate experimental sessions performed
1 week apart. SW was computed to evaluate brain network organization. The present study demonstrates that tDCS delivered in
PFC can change brain network dynamics, and tDCS-EEG coregistration data can be analyzed using graph theory to understand
the induced effects of different tDCS polarities in physiological and pathological brain aging.

Keywords Graph theory-EEG . Aging . Functional connectivity . Rehabilitation . tDCS

Introduction non-invasive brain stimulation techniques have attracted wide

In the last years, several efforts have been performed to amelio-
rate cognitive functions and behavioral outcomes in physiologi-
cal aging with “neuroenhancement” approaches [6]. Recently,
This article is part of the special issue on Aging Brain in Pflügers
Archiv—European Journal of Physiology
* Fabrizio Vecchio
fabrizio.vecchio@uniroma1.it; fabrizio.vecchio@sanraffaele.it
1 ing spontaneous neuronal firing rate and the cortical excitabil-
Brain Connectivity Laboratory, Department of Neuroscience &
Neurorehabilitation, IRCCS San Raffaele Pisana, Via Val Cannuta,
247, 00166 Rome, Italy
2
Cognitive Neuroscience Section, IRCCS Istituto Centro San
Giovanni di Dio, Fatebenefratelli, Brescia, Italy
3
Center for Mind/Brain Sciences – CIMeC, University of Trento,
Rovereto, TN, Italy
interest for their ability to modify functional neural activation
patterns and the related cognitive functions in the aging brain
[26]. Among the neuromodulation approaches, transcranial direct
current stimulation (tDCS) represents an additional candidate
tool for maintaining and strengthening cognitive functioning of
the older adults, by promoting neuronal changes in brain activity
and connectivity [6].
Regardless of the polarity, tDCS affects neuronal excitabil-
ity by delivering weak currents to cerebral tissues by means
of, at least, two electrodes. In particular, anodal tDCS depo-
larizes the resting membrane potential of the neuron enhanc-
ity increase, in the stimulated area, while cathodal tDCS
causes a shift of the resting membrane potential towards hy-
perpolarization, thus decreasing the excitability of the cortex
and reducing the neuronal firing [8].
Albeit recent studies have adopted this technique to mod-
ulate neuronal activity underlying cognitive functions both in

Content courtesy of Springer Nature, terms of use apply. Rights reserved.

786
young and older adults [5, 25], the tDCS-induced effects at
network level in physiological aging brain remains to be de-
fined. A better understanding of the neural changes induced
by tDCS at cortical network level should precede the assess-
ment of its effects at behavioral and cognitive levels [2].
Starting from the well-established role of the prefrontal cortex
(PFC) in executive functions, working memory abilities, and
speed of information processing, PFC could be considered an
ideal cortical target for evaluating the capability of tDCS to induce
functional connectivity changes in brain aging.
Considering that oscillatory activities in neural systems
play a key role in orchestrating brain functions and that
tDCS may be used to interact with brain oscillations and af-
fects in polarity-dependent manner specific cortical connec-
tivity patterns [14, 29], we used a network approach to eval-
uate age-dependent brain connectivity changes induced by
different tDCS polarities.
At general level, recent methods based on network science
were developed to evaluate brain connectivity changes. The
brain tends to be modeled as a complex combination of net-
works by the network science; in mathematical words, a net-
work is defined by a set of nodes and links between pairs of
nodes. Physiologically, the brain regions could be represented
by the nodes, while the connections by the links [9] and the
connections that can be established between neuronal assem-
blies are the result of segregation and integration processes, as
mathematically revealed by local clustering (segregation) and
path length (integration). Usually, a high degree of local clus-
tering (segregation) and long-distance connections
(integration) characterizes the network topology of the cere-
bral connections. A model of network organization, the
“Small World” (SW) concept, was introduced allowing for
an optimal balance between global integration and local spe-
cialization [32]. The architecture at the base of the brain func-
tional connectivity could be modeled by means of this ap-
proach [4], in order to correlate it with behavior (i.e., task-
performance). This helps to evaluate if patterns of functional
connectivity between brain regions reflect the organization of
more-or-less strongly connected networks based on the phase
coherence of oscillatory firing synchronizations between
adjacent/remote neuronal assemblies, as shown by their EEG
activity in a milliseconds time frame [7, 28].
Within this theoretical framework, the present study aims
to systematically investigate the tDCS-induced effects on
brain network properties during physiological aging. At this
aim, EEG activity was acquired from healthy young and older
adult participants before, during, and after anodal, cathodal,
and sham tDCS applied over PFC. In order to explore online
polarity-dependent changes in functional connectivity, a net-
work approach was used to assess how the tDCS affects the
brain network, by the evaluation of the SW parameter.
Specifically, our aim to explore tDCS polarity-dependent
changes in the network dynamics during aging was based on
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Pflugers Arch - Eur J Physiol (2021) 473:785–792
a network graph theory application on two groups of young
and elderly participants.
Material and methods
Participants
Eighteen healthy young (9 females; mean age = 24.7 years,
SD = 3.2) and fifteen elderly adult participants (9 females;
mean age = 70.1 years, SD = 5.1) participated in the present
study. All recruited subjects were right-handed, as evaluated
using the Edinburgh handedness inventory test (Oldfield,
1971). Moreover, all participants reported no previous history
of neurological or psychiatric disorders and had no metal im-
plants. Individual written informed consent was obtained, and
the study was approved by a local ethical committee.
Experimental procedures conformed to the Declaration of
Helsinki and received prior approval by the Ethical
Committee of IRCCS Istituto Centro San Giovanni di Dio,
Fatebenefratelli. The tDCS protocols were performed in ac-
cordance with safety guidelines procedures [1].
Data and code availability statement
The data adopted by the authors comply with the requirements
of the institute and comply with institutional ethics approval.
The data that support the findings of this study are available
from the corresponding author on request.
Experimental design
Each participant received anodal, cathodal, or sham tDCS
over the left PFC in three separate experimental sessions per-
formed 1 week apart. The order of tDCS sessions was ran-
domized and counterbalanced among participants. During
each session, the EEG signals were recorded before (5 min)
and after (5 min) tDCS, whereas an additional EEG recording
was performed for the entire tDCS session (13 min). During
the experiment, the participants were seated on a dedicated,
comfortable armchair in a Faraday-cage, sound-proofed room.
Moreover, they were instructed to keep their eyes open, avoid
blinking, and to look at a stationary fixed point in the center of
a computer screen. The participants were blind to the tDCS
conditions. Figure 1 shows the experimental protocol.
EEG recordings and preprocessing
Electroencephalographic recordings were performed with a
standard montage (Easycap, GmbH, Brain Products) from
31 electrodes positioned in accordance with the 10–20
International System (Fp1, Fp2, AF7, AF8, F7, F3, Fz, F4,
F8, FC5, FC1, FC2, FC6, T7, C3, Cz, C4, T8, CP5, CP1, CP2,
Pflugers Arch - Eur J Physiol (2021) 473:785–792
Fig. 1 Experimental protocol
CP6, P7, P3, Pz, P4, P8, PO7, PO8, O1, O2), acquired with a
0.1–1000-Hz band-pass filter and digitized at a 5000-Hz sam-
pling rate. The midfrontal (Fpz) electrode was used as the
ground one. The online reference for all electrodes was the
right mastoid electrode (M2), while the left mastoid electrode
(M1) was used only offline in order to re-reference the record-
ings to the average of the two mastoids. Two different chan-
nels with vertical and horizontal montages monitored the eye
movement. The impedances between skin and electrode were
lowered below 5 KΩ.
Data were analyzed by means of scripts from EEGLAB
(Swartz Center for Computational Neurosciences, La Jolla,
CA), a toolbox of Matlab (Math Works, Natick, MA) soft-
ware. The EEG signals were band-pass filtered from 0.2 to
47 Hz through a finite impulse response (FIR) filter and down-
sampled to 512 Hz sampling rate. The imported data were
fragmented in 2-s duration epochs, and muscular, cardiac,
ocular, and other types of artifacts were inspected. The proce-
dure was the following: (1) an EEG expert reviewed the data,
discarding manually the epochs with evident artifactual activ-
ity or with aberrant waveforms, and (2) an independent com-
ponent analysis (ICA) was lastly performed on the 2-s epochs
to complete the detection and rejection of artifacts using the
Infomax ICA algorithm, which is implemented in the
EEGLAB. ICA is a blind source decomposition algorithm
which enables to separate statistically independent sources
from data acquired from multiple channels. This method is
considered as particularly efficient in separating blink and
ocular movement artifacts from EEG data. The EEG expert
visually inspected the components and, if artifact contamina-
tion was found, they were manually rejected.
tDCS
The direct current stimulation was delivered by a battery-
driven electrical stimulator (Brain Stim, EMS, Bologna,
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
787
Italy) through two conductive-rubber electrodes. The active
(anodal or cathodal) electrode (area:16 cm2) was placed over
the left PFC (in correspondence with F3 position on the EEG
cap), whereas the other electrode (50 cm2) was fixed extra-
cephalically on the right arm. For the active condition, the
current was delivered with an intensity of 1.5 mA for
13 min, with a ramping period of 8 s both at the beginning
and at the end of the stimulation. For sham tDCS, the current
was delivered only for 10 s at the beginning and at the end of
the stimulation time. The terms “anodal” and “cathodal” refer
to the polarity of the electrode placed over the left PFC.
Functional connectivity analysis
The exact low-resolution electromagnetic tomography
(eLORETA) software was used to carry out EEG functional
connectivity analysis [15, 16, 27, 28]. The eLORETA algo-
rithm is a linear inverse solution for EEG data without local-
ization error which is able to provide sources under ideal
(noise-free) conditions [18]. eLORETA software, in accor-
dance with the EEG potential distribution of the scalp, was
used to compute a discrete, three-dimensionally (3D) distrib-
uted linear, minimum-norm, weighted, inverse solution. The
weights used in eLORETA enable to localize exactly the
sources, a property that is necessary to test point sources,
yielding images of the current density with exact localization,
even if with a low spatial resolution (i.e., there is a high cor-
relation between neighboring neuronal sources).
Brain connectivity was evaluated by means of eLORETA
software in 84 regions to obtain a topographic view of the
entire brain, placing the center in the available 42 Brodmann
areas (BAs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 17, 18, 19, 20,
21, 22, 23, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,
39, 40, 41, 42, 43, 44, 45, 46, 47) in the left and right cerebral
hemispheres.
788
To estimate the electrical brain activity that is used to in-
vestigate cerebral-functional connectivity, the regions of inter-
est (ROIs) are needed. The electric neuronal activities of all
voxels belonging to each cortical ROI are averaged in order to
get a single signal for each of them, as computed with
eLORETA. For each brain hemisphere, the intracortical
lagged linear coherence among the eLORETA current density
time series of the 84 ROIs, extracted via the method of “all
nearest voxels” [19], was evaluated between all possible pairs
of the 84 ROIs for all the seven independent EEG frequency
bands of delta, theta, alpha 1, alpha 2, beta 1, beta 2, and
gamma (2–4 Hz; 4–8 Hz; 8–10.5 Hz; 10.5–13 Hz; 13–
20 Hz; 20–30 Hz; 30–45 Hz respectively) for each condition
and participant.
Considering the definition of the complex valued coher-
ence between the two time series x and y in the frequency
band ω—that is based on the cross-spectrum given by the
variance and the covariance of the signals—the lagged linear
coherence computed in the frequency band ω is computed
according to the equation that follows [19]:
½ImCovðx; yÞ2
LagR2xyw ¼
VarðxÞ*VarðyÞ−½ReCovðx; yÞ2
where CoV and Var are the covariance and variance respec-
tively of the time series.
This parameter provides a measure of true physiological
cerebral connectivity not influenced by low spatial resolution
and volume conduction [19]. The lagged linear connectivity
values computed for each frequency band between all pairs of
ROIs constitute the weights of the networks built in the graph
analysis.
Graph analysis
As already defined, a real-world complex system is character-
ized by a series of nodes and links between pairs of nodes
(vertices and edges). Cerebral regions usually are represented
by nodes, while functional, anatomical, or effective connec-
tions [9], depending on the dataset, are links. Normally, ana-
tomical connections correspond to white matter fiber tracts
between couples of gray matter brain areas (cortical regions
or subcortical relays). Functional connections reflect the mag-
nitudes of correlations in activity which occur temporarily and
which may occur even between pairs of regions that are not
structurally connected directly.
A group of elements (vertices) that may be linked by means
of connections of variable weights (edges) constitutes, from a
mathematical point of view, a weighted graph.
In this study, the undirected and weighted networks were
built (the estimated cortical sources in the BAs constitute the
network vertices) and the edges were weighted by the lagged
linear value within each couple of vertices. The graph analysis
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Pflugers Arch - Eur J Physiol (2021) 473:785–792
was carried out using the Brain Connectivity Toolbox (BCT,
brain-connectivity-toolbox.net), a software instrument that
was adapted with the Matlab scripts developed in our
laboratory.
On the brain networks, the SW parameter was evaluated,
since it measures the balance between global integration and
local connectedness of a network, providing a picture of brain
network organization. The ratio of the normalized clustering
coefficient and the normalized path length constitutes the
small-worldness measure [22]. Before performing the small-
world measurements, a data normalization (i.e., relativization)
was applied. The clustering coefficient and the characteristic
path length values were divided by the mean ones obtained by
the average measure of each parameter in the EEG frequency
bands of each participant, in order to obtain individual mea-
sures. As we computed from weighted networks, it was com-
plicated to evaluate disgraphs with the same number of con-
nections (all connections were available) and nodes; thus, we
decided to employ relative values within each frequency band
[3, 21, 30].
Statistical evaluation
Once extracted a graph pattern with eLORETA from the brain
network, the eLORETA statistical evaluation was performed.
The Kolmogorov-Smirnov test was used to verify that data
normality, and the Gaussianity hypothesis could not be
rejected. A statistical ANOVA design for each group of sub-
jects (Young and Older adults) was performed for the Small
World between the three factors: stimulation (anodal, cathod-
al, sham), time (pre, during, post), and frequency band (delta,
theta, alpha 1, alpha 2, beta 1, beta 2, and gamma) in order to
validate the working hypothesis.
Finally, in order to address a direct match between the two
group of participants, a statistical analysis was evaluated in-
cluding group of participants (Young and Older adults) as
between factor and stimulation (anodal, cathodal) in the band
resulted statistically different in the previous analysis.
Furthermore, correlation analyses were carried out consider-
ing all participants as a whole group separately for the fre-
quency bands presenting statistical differences between the
two groups of participants in each kind of stimulation.
Theory
Brain neural networks undergo relevant changes during phys-
iological aging, which affect cognitive and behavioral func-
tions. Currently, non-invasive brain stimulation techniques,
such as transcranial direct current stimulation (tDCS), are
adopted to modulate cognitive functions in brain aging, acting
on networks properties and connectivity. This could be used
to address rehabilitation treatment not only in physiological
aging but also in pathological brain aging.
Pflugers Arch - Eur J Physiol (2021) 473:785–792
Results
Graph theory parameter analyses
Both the ANOVAs (Fig. 2) for the evaluation of the SW in
Young and Older adults showed statistically significant inter-
actions (young: F(24, 408) = 2.2962, p = 0.00057; older
adults: F(24,360) = 2.8435; p < 0.00002) between stimulation
(anodal, cathodal, sham), time (pre, during, post), and fre-
quency band (delta, theta, alpha 1, alpha 2, beta 1, beta 2,
and gamma) factors.
The post hoc tests showed no significant differences in pre
and post condition, probably also due to the long time distance
passed after stimulation [31], while several differences were
found during the stimulation, as reported in the following
paragraphs.
Fig. 2 ANOVAs for the
evaluation of the SW in young
and older adults
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
789
Sham stimulation vs real stimulations
In young, Duncan-planned post hoc testing showed lower
values in theta (p < 0.000022) band, and higher values in al-
pha 2 band (p < 0.008954) during anodal tDCS with respect to
sham stimulation. Duncan-planned post hoc testing showed
also lower values in delta (p < 0.000713) and theta
(p < 0.000039) bands and higher values in alpha 1
(p < 0.005393) and alpha 2 bands (p < 0.001567) during cath-
odal tDCS with respect to sham stimulation.
While considering older adults, Duncan-planned post hoc
testing showed lower values in delta (p < 0.001617) and theta
(p < 0.028757) bands and higher values in alpha 2 band
(p < 0.003434) during anodal tDCS with respect to sham stim-
ulation. Duncan-planned post hoc testing showed also lower
values in delta (p < 0.000308) band and higher values in alpha
790
1 (p < 0.016822) and alpha 2 bands (p < 0.023037) during
cathodal tDCS with respect to sham stimulation.
Anodal stimulation vs cathodal stimulation
In young, Duncan-planned post hoc testing showed higher
values in alpha 1 band (p < 0.011404) during cathodal with
respect to anodal tDCS. While considering older adults,
Duncan-planned post hoc testing showed no differences be-
tween anodal and cathodal stimulation.
In order to address a direct match among age, a statistical
analysis was evaluated including group of participants (young
and older adults, as between factor) and stimulation (anodal,
cathodal) in the alpha 1 band. The result showed a statistically
significant interactions (F(1, 32) = 4.0169, p = 0.049) and
Duncan-planned post hoc testing showing higher values in
cathodal stimulation for young with respect to older subjects
(p < 0.002797).
Finally, considering all participants as a whole group,
correlation analysis on alpha 1 band showed a negative
correlation with age (p = 0.0368, r = − 0.3706), namely,
higher the age lower the alpha 1 values during cathodal
stimulation.
Summarizing, elderly participants present different effects
regarding real tDCS with respect to sham stimulation but no
specific changes induced by different tDCS polarities.
Otherwise, young participants present different effects with
respect to tDCS polarity.
Functional coupling evaluation
Figure 3 reports the functional coupling distribution, as
revealed by the lagged linear coherence, in the above
significant EEG frequency bands (delta, theta, alpha 1,
alpha 2) in the two groups of participants during tDCS
period in each stimulation.
Fig. 3 Functional coupling. An
arbitrary threshold was used to
illustrate these patterns
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Pflugers Arch - Eur J Physiol (2021) 473:785–792
Discussion
The typical organization of a healthy brain is characterized by
an optimal balance of functional segregation and integration;
SW is the term used to define this kind of scenario. The intri-
cate inhibitory and excitatory cerebral networks consist of
areas which are functionally specialized, and that reciprocally
and continuously cooperate to acquire, share, and integrate
any kind of information; all of this reflects the SW features
and happens in a constant state of dynamic fluctuations. Our
study demonstrates that tDCS delivered over PFC can change
ongoing network dynamics, in terms of SW properties.
Additionally, our findings support the idea that the graph the-
ory can be properly used for analyzing tDCS-EEG
coregistration data with the purpose to understand the effects
induced by different tDCS polarities including in exploring
age-related brain changes.
Specifically, our results highlight a statistically significant dif-
ference in the SW organization during real tDCS respect to sham
stimulation both in young and older adult participants while spe-
cific frequency differences during cathodal with respect to anodal
stimulation were found only in young participants.
Particularly, a SW increase in alpha and a reduction in low
(delta and theta) frequencies for both groups of subjects were
found for real tDCS respect to sham stimulation. This effect
should be explained keeping in mind the physiological role
that the alpha rhythm plays. The waking rest EEG activity is
characterized by alpha frequencies, usually defined as the
“idling rhythms” of the adult brain [17]. Alpha has been
proved by several studies to be a brain rhythm involved in
several cerebral functions, ranging from sensory-motor pro-
cessing to memory formation [23]. Alpha rhythm constitutes
the base of information transmission in healthy subject brains,
because it works as an oscillatory component of cerebral ac-
tivity [11]. Furthermore, studies dealing with event related
EEG have found that alpha rhythm is positively correlated
with the information processing speed and with a good cog-
nitive performance [11].
Pflugers Arch - Eur J Physiol (2021) 473:785–792
Regarding the low-frequency bands, it is affirmed that,
in a waking state, these EEG rhythms are poorly repre-
sented, as if alpha and delta rhythms are in a state of
“reciprocal inhibition” [21]. Furthermore, it is well
known that spontaneous oscillations in the delta frequen-
cies in almost all recorded neurons are generated in case
of functional or anatomical disconnection of damaged
cortical regions [12]. The found SW decrease in the
low-frequency bands could reflect a sort of structured
behavior consisted of a functional inhibition and an in-
crease in activity. The opposite is true for the alpha band.
Finally, older adults present different effects with respect to
sham tDCS but no specific effects for the polarity of stimula-
tion. Otherwise, young participants present different effects
with respect to the tDCS polarities: cathodal stimulation pro-
vides higher effect in alpha 1 than anodal one, namely, higher
SW values in cathodal are observed. This finding is strongly
in line with the non-linear effects of cathodal tDCS recently
reported [24]. The high level of alpha 1 SW was also found
negatively correlated with the age of participants, namely, the
higher the age, the lower the SW values in alpha 1 during
cathodal stimulation. The specificity of alpha effect in young
participants during cathodal stimulation could be seen as an
increase in activity and a functional excitation no more present
in aging brain; a mechanism of brain reactivity is still present
in young participants.
Albeit recent studies failed to show tDCS polarity-
dependent changes in resting-state EEG highlighting the lack
of specific after effects on cortical activity as evaluated by
spectral power analysis [10], our findings about the tDCS-
induced effects at network level are in line with previous re-
sults on inter-hemispheric changes [20] and resting-state brain
connectivity modulations after tDCS [13], highlighting also
that the polarity-specific effects of tDCS on cortical connec-
tivity at rest could be partly age-dependent.
As general remarks, further studies could be carried out by
non-linear techniques and also to look at other graph parame-
ters such as centrality, modularity, hub, sub graphs, and so on
to see for example if there are any isolated clusters to be more
modulated by the stimulations.
In conclusion, our results underline that tDCS affects the
brain organization of specific functional networks at specific
frequency bands in a different manner related to the subject
age. This might be used to address rehabilitation treatment not
only in physiological aging but also in pathological brain ag-
ing in a way that needs further studies. Furthermore, since
resting-state patterns of brain connectivity are the result of
robust and specific intrinsic neural activity, our results high-
light that tDCS is able to affect the ongoing spontaneous neu-
ronal fluctuations; this seems particularly interesting in view
of age-related changes and the modulation of the dynamics of
the brain at rest to study specific effects in motor or mental
activities.
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
791
Funding information This work was partially supported by the Italian
Ministry of Health for Institutional Research (Ricerca corrente) and for
the project GR-2011-02349998.
Data availability The data will be available upon request.
Compliance with ethical standards The data adopted by the
authors comply with the requirements of the institute and comply with
institutional ethics approval.
Conflict of interest The authors declare that they have no conflict of
interest.
References
1. Antal A, Alekseichuk I, Bikson M, Brockmöller J, Brunoni AR,
Chen R, Cohen LG, Dowthwaite G, Ellrich J, Flöel A, Fregni F,
George MS, Hamilton R, Haueisen J, Herrmann CS, Hummel FC,
Lefaucheur JP, Liebetanz D, Loo CK, McCaig CD, Miniussi C,
Miranda PC, Moliadze V, Nitsche MA, Nowak R, Padberg F,
Pascual-Leone A, Poppendieck W, Priori A, Rossi S, Rossini PM,
Rothwell J, Rueger MA, Ruffini G, Schellhorn K, Siebner HR,
Ugawa Y, Wexler A, Ziemann U, Hallett M, Paulus W (2017)
Low intensity transcranial electric stimulation: safety, ethical, legal
regulatory and application guidelines. Clin Neurophysiol 128:
1774–1809. https://doi.org/10.1016/j.clinph.2017.06.001
2. Antonenko D, Schubert F, Bohm F, Ittermann B, Aydin S, Hayek
D, Grittner U, Flöel A (2017) tDCS-induced modulation of GABA
levels and resting-state functional connectivity in older adults. J
Neurosci 37:4065–4073. https://doi.org/10.1523/JNEUROSCI.
0079-17.2017
3. Babiloni C, Vecchio F, Lizio R, Ferri R, Rodriguez G, Marzano N,
Frisoni GB, Rossini PM (2011) Resting state cortical rhythms in
mild cognitive impairment and Alzheimer’s disease: electroenceph-
alographic evidence. J Alzheimers Dis 26(Suppl 3):201–214.
https://doi.org/10.3233/jad-2011-0051
4. Bassett DS, Bullmore E (2006) Small-world brain networks.
Neu roscie ntis t 1 2:5 12 –52 3. ht tps: //doi.org/10.1177/
1073858406293182
5. Cespón J, Rodella C, Rossini PM, Miniussi C, Pellicciari MC
(2017) Anodal transcranial direct current stimulation promotes
frontal compensatory mechanisms in healthy elderly subjects.
Front Aging Neurosci 9:420. https://doi.org/10.3389/fnagi.2017.
00420
6. Cespón J, Miniussi C, Pellicciari MC (2018) Interventional
programmes to improve cognition during healthy and pathological
ageing: cortical modulations and evidence for brain plasticity.
Ageing Res Rev 43:81–98. https://doi.org/10.1016/j.arr.2018.03.
001
7. de Haan W, Pijnenburg YA, Strijers RL, van der Made Y, van der
Flier WM, Scheltens P, Stam CJ (2009) Functional neural network
analysis in frontotemporal dementia and Alzheimer’s disease using
EEG and graph theory. BMC Neurosci 10:101. https://doi.org/10.
1186/1471-2202-10-101
8. Fertonani A, Miniussi C (2017) Transcranial electrical stimulation:
what we know and do not know about mechanisms. Neuroscientist
23:109–123. https://doi.org/10.1177/1073858416631966
9. Friston KJ (2004) Functional and effective connectivity in neuro-
imaging: a synthesis. Hum Brain Mapp 2:56–78. https://doi.org/10.
1002/hbm.460020107
10. Gordon PC, Zrenner C, Desideri D, Belardinelli P, Zrenner B,
Brunoni AR, Ziemann U (2018) Modulation of cortical responses
by transcranial direct current stimulation of dorsolateral prefrontal
792
cortex: a resting-state EEG and TMS-EEG study. Brain Stimul 11:
1024–1032. https://doi.org/10.1016/j.brs.2018.06.004
11. Klimesch W (1999) EEG alpha and theta oscillations reflect cogni-
tive and memory performance: a review and analysis. Brain Res
Brain Res Rev 29:169–195
12. Kooi KA (1922) Fundamentals of electroencephalography.
Medical Dept., Harper & Row, New York
13. Luft CD, Pereda E, Banissy MJ, Bhattacharya J (2014) Best of both
worlds: promise of combining brain stimulation and brain
connectome. Front Syst Neurosci 8:132. https://doi.org/10.3389/
fnsys.2014.00132
14. Mancini M, Brignani D, Conforto S, Mauri P, Miniussi C,
Pellicciari MC (2016) Assessing cortical synchronization during
transcranial direct current stimulation: a graph-theoretical analysis.
Neuroimage 140:57–65. https://doi.org/10.1016/j.neuroimage.
2016.06.003
15. Miraglia F, Vecchio F, Bramanti P, Rossini PM (2016) EEG char-
acteristics in “eyes-open” versus “eyes-closed” conditions: small-
world network architecture in healthy aging and age-related brain
degeneration. Clin Neurophysiol 127:1261–1268. https://doi.org/
10.1016/j.clinph.2015.07.040
16. Miraglia F, Vecchio F, Rossini PM (2017) Searching for signs of
aging and dementia in EEG through network analysis. Behav Brain
Res 317:292–300. https://doi.org/10.1016/j.bbr.2016.09.057
17. Niedermeyer E, Lopes da Silva F (1993) Electroencephalography,
basic principles, clinical applications and related fields. Oxford
University Press, Oxford
18. Pascual-Marqui RD (2002) Standardized low-resolution brain elec-
tromagnetic tomography (sLORETA): technical details. Methods
Find Exp Clin Pharmacol 24(Suppl D):5–12
19. Pascual-Marqui RD (2007) Instantaneous and lagged measure-
ments of linear and nonlinear dependence between groups of mul-
tivariate time series: frequency decomposition. eprint arXiv:
0711.1455
20. Pellicciari MC, Brignani D, Miniussi C (2013) Excitability modu-
lation of the motor system induced by transcranial direct current
stimulation: a multimodal approach. Neuroimage 83:569–580.
https://doi.org/10.1016/j.neuroimage.2013.06.076
21. Rossini PM, Del Percio C, Pasqualetti P, Cassetta E, Binetti G, Dal
Forno G, Ferreri F, Frisoni G, Chiovenda P, Miniussi C, Parisi L,
Tombini M, Vecchio F, Babiloni C (2006) Conversion from mild
cognitive impairment to Alzheimer’s disease is predicted by
sources and coherence of brain electroencephalography rhythms.
Neuroscience 143:793–803. https://doi.org/10.1016/j.
neuroscience.2006.08.049
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Pflugers Arch - Eur J Physiol (2021) 473:785–792
22. Rubinov M, Sporns O (2010) Complex network measures of brain
connectivity: uses and interpretations. Neuroimage 52:1059–1069.
https://doi.org/10.1016/j.neuroimage.2009.10.003
23. Schürmann M, Başar E (2001) Functional aspects of alpha oscilla-
tions in the EEG. Int J Psychophysiol 39:151–158
24. Shilo G, Lavidor M (2019) Non-linear effects of cathodal transcra-
nial direct current stimulation (tDCS) of the primary motor cortex
on implicit motor learning. Exp Brain Res 237:919–925. https://doi.
org/10.1007/s00221-019-05477-3
25. Summers JJ, Kang N, Cauraugh JH (2016) Does transcranial direct
current stimulation enhance cognitive and motor functions in the
ageing brain? A systematic review and meta- analysis. Ageing Res
Rev 25:42–54. https://doi.org/10.1016/j.arr.2015.11.004
26. Tatti E, Rossi S, Innocenti I, Rossi A, Santarnecchi E (2016) Non-
invasive brain stimulation of the aging brain: state of the art and
future perspectives. Ageing Res Rev 29:66–89. https://doi.org/10.
1016/j.arr.2016.05.006
27. Vecchio F, Miraglia F, Bramanti P, Rossini PM (2014) Human
brain networks in physiological aging: a graph theoretical analysis
of cortical connectivity from EEG data. J Alzheimers Dis 41:1239–
1249. https://doi.org/10.3233/JAD-140090
28. Vecchio F, Miraglia F, Marra C, Quaranta D, Vita MG, Bramanti P,
Rossini PM (2014) Human brain networks in cognitive decline: a
graph theoretical analysis of cortical connectivity from EEG data. J
Alzheimers Dis 41:113–127. https://doi.org/10.3233/jad-132087
29. Vecchio F, Pellicciari MC, Miraglia F, Brignani D, Miniussi C,
Rossini PM (2016) Effects of transcranial direct current stimulation
on the functional coupling of the sensorimotor cortical network.
Neuroimage 140:50–56. https://doi.org/10.1016/j.neuroimage.
2016.01.051
30. Vecchio F, Miraglia F, Piludu F, Granata G, Romanello R, Caulo
M, Onofrj V, Bramanti P, Colosimo C, Rossini PM (2017) “Small
world” architecture in brain connectivity and hippocampal volume
in Alzheimer’s disease: a study via graph theory from EEG data.
Brain Imaging Behav 11:473–485. https://doi.org/10.1007/s11682-
016-9528-3
31. Vecchio F, Di Iorio R, Miraglia F, Granata G, Romanello R,
Bramanti P, Rossini PM (2018) Transcranial direct current stimu-
lation generates a transient increase of small-world in brain connec-
tivity: an EEG graph theoretical analysis. Exp Brain Res 236:1117–
1127. https://doi.org/10.1007/s00221-018-5200-z
32. Watts DJ, Strogatz SH (1998) Collective dynamics of ‘small-world’
networks. Nature 393:440–442. https://doi.org/10.1038/30918
Publisher’s note Springer Nature remains neutral with regard to jurisdic-
tional claims in published maps and institutional affiliations.
Terms and Conditions
Springer Nature journal content, brought to you courtesy of Springer Nature Customer Service Center GmbH (“Springer Nature”).
Springer Nature supports a reasonable amount of sharing of research papers by authors, subscribers and authorised users (“Users”), for small-
scale personal, non-commercial use provided that all copyright, trade and service marks and other proprietary notices are maintained. By
accessing, sharing, receiving or otherwise using the Springer Nature journal content you agree to these terms of use (“Terms”). For these
purposes, Springer Nature considers academic use (by researchers and students) to be non-commercial.
These Terms are supplementary and will apply in addition to any applicable website terms and conditions, a relevant site licence or a personal
subscription. These Terms will prevail over any conflict or ambiguity with regards to the relevant terms, a site licence or a personal subscription
(to the extent of the conflict or ambiguity only). For Creative Commons-licensed articles, the terms of the Creative Commons license used will
apply.
We collect and use personal data to provide access to the Springer Nature journal content. We may also use these personal data internally within
ResearchGate and Springer Nature and as agreed share it, in an anonymised way, for purposes of tracking, analysis and reporting. We will not
otherwise disclose your personal data outside the ResearchGate or the Springer Nature group of companies unless we have your permission as
detailed in the Privacy Policy.
While Users may use the Springer Nature journal content for small scale, personal non-commercial use, it is important to note that Users may
not:

1. use such content for the purpose of providing other users with access on a regular or large scale basis or as a means to circumvent access
control;
2. use such content where to do so would be considered a criminal or statutory offence in any jurisdiction, or gives rise to civil liability, or is
otherwise unlawful;
3. falsely or misleadingly imply or suggest endorsement, approval , sponsorship, or association unless explicitly agreed to by Springer Nature in
writing;
4. use bots or other automated methods to access the content or redirect messages
5. override any security feature or exclusionary protocol; or
6. share the content in order to create substitute for Springer Nature products or services or a systematic database of Springer Nature journal
content.
In line with the restriction against commercial use, Springer Nature does not permit the creation of a product or service that creates revenue,
royalties, rent or income from our content or its inclusion as part of a paid for service or for other commercial gain. Springer Nature journal
content cannot be used for inter-library loans and librarians may not upload Springer Nature journal content on a large scale into their, or any
other, institutional repository.
These terms of use are reviewed regularly and may be amended at any time. Springer Nature is not obligated to publish any information or
content on this website and may remove it or features or functionality at our sole discretion, at any time with or without notice. Springer Nature
may revoke this licence to you at any time and remove access to any copies of the Springer Nature journal content which have been saved.
To the fullest extent permitted by law, Springer Nature makes no warranties, representations or guarantees to Users, either express or implied
with respect to the Springer nature journal content and all parties disclaim and waive any implied warranties or warranties imposed by law,
including merchantability or fitness for any particular purpose.
Please note that these rights do not automatically extend to content, data or other material published by Springer Nature that may be licensed
from third parties.
If you would like to use or distribute our Springer Nature journal content to a wider audience or on a regular basis or in any other manner not
expressly permitted by these Terms, please contact Springer Nature at

onlineservice@springernature.com