Early Child Development and Care

Vol. 176, Nos 3&4, May 2006, pp. 285–298

Piracetam: its possible mode of action

in children with learning disabilities and
its effect on in vitro cell growth
R. Britz, M. J. Bester, A. da Silva, N. A. Motsoane, J. Marx,
H. Naudé and E. Pretorius*
University of Pretoria, South Africa
Taylor and Francis Ltd
GECD041078.sgm

(Received 18 June 2004)

Early
10.1080/0300443042000302672
0300-4430
Original
Taylor
02005
Faculty
E.Pretorius
000002005
Child
&ofArticle
Francis
Health
(print)/1476-8275
Development
Group
SciencesUniversity
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and(online)
Care of PretoriaBMW BuildingPO Box 2034Pretoria 0001South Africaresia.pretorius@up.ac.za

The use of pharmaceutical products such as Piracetam (Nootropil®) for the treatment of learning
disabilities is becoming increasingly prevalent, and some studies have shown successful treatment
of learning disabilities in children. This research article will discuss traditional uses of Piracetam, as
well as uses in learning disabilities, with particular focus on case studies where it has been given to
children with learning disabilities. Furthermore, we investigate how Piracetam could potentially
enhance learning, with specific focus on brain areas. However, even though Piracetam may enhance
learning, the question that arises is how safe it is at a cellular level. Therefore the effect of Piracetam
at a cellular level was determined. To achieve this purpose the effect of a 24-hour exposure to 0–70
mM Piracetam on the growth of the L929 cell line and chick embryonic neurons in vitro was inves-
tigated. No statistically significant changes in cell viability or number was observed at 0–7 mM
Piracetam in both cell types, although at 70 mM cell viability in the L929 cell line was significantly
reduced when compared with the control. Initial studies indicate that Piacetam is only toxic at
concentrations greater than 70 mM; however, the effects of repeated dosages over long periods still
need to be investigated.

Keywords: Cells in culture; Learning disabilities; Nootropil®; Pharmaceutical products

Introduction
Worldwide learning disabilities, severe or mild, affect many children as well as adults.
How to positively enhance learning experiences of learners suffering from these prob-
lems has been the target of investigations by numerous researchers. Several approaches

*Corresponding author. BMW Building, PO Box 2034, Faculty of Health Sciences, University of
Pretoria, Pretoria 0001, South Africa. Email: resia.pretorius@up.ac.za

ISSN 0300-4430 (print)/ISSN 1476-8275 (online)/06/030285–14

© 2006 Taylor & Francis
DOI: 10.1080/03004430500258461
286 R. Britz et al.

for the management of learning disabilities are used. One approach is where problem
areas are identified and then the specific problem is addressed by determining weak-
nesses and strengths. Furthermore, metacognitive skills may be used to address the
problem. Alternatively, by establishing which brain areas are involved in the disability,
and then focusing on areas not affected to enhance the learning capabilities, can also
be utilized. A question that often arises is how safe it is to use pharmaceutical products
in addressing learning disabilities. One such product is Nootropil®, otherwise known
as Piracetam, that was introduced to the scientific community in the late 1960s. Over
the years it has been used to treat various cerebral disorders and a few researchers have
used it with great success to treat learning disabilities in children (these results were
based on case studies).
This research article will discuss traditional uses of Piracetam, as well as uses in
learning disabilities, with particular focus on case studies where it has been given to
children with learning disabilities. Furthermore, we investigate how Piracetam could
potentially enhance learning, with specific focus on brain areas. However, even
though Piracetam may enhance learning, the question that arises is how safe is it at a
cellular level. Therefore the effect of Piracetam at a cellular level will be determined,
by investigating its effect on fibroblast and neuron number and viability in vitro.

Literature review
Piracetam: modes of action and its traditional uses
Modes of action. Piracetam (2-oxo-1-pyrrolidine acetamide) is derived from
(gamma)-aminobutyric acid (Vastag et al., 1999; Kessler et al., 2000). It has a low
molecular weight, is highly soluble in water and only partially soluble in organic
media. It is not metabolized in the body, nor is it ionized or bound to proteins
(Paulesu et al., 2001). Furthermore, it appears to be neither stimulating nor sedating,
but acts as a general cortical activator that induces alertness (Wilsher et al., 1985).
Traditionally Piracetam has been used as a neuro-protective and rehabilitating drug,
and it is believed to have a neurometabolic effect (Sidorov et al., 2000), as well as a
cyto-protective effect on brain tissue, and it improves cerebral blood flow (Gociski
et al., 1999).
Moriau et al. (1993) mentioned that there are four sites of action of Piracetam,
namely the vessel wall, platelets, plasma and cell membranes; this provides the basis
for the potentially important antithrombotic activity of the product. Kawiak et al.
(1995) also mentioned that Piracetam exerts a positive impact on both intracortical
and cortico-subcortical conduction and increases ATP content in the nerve cell,
which leads to the increase of ribonucleic acid synthesis.
Piracetam also promotes erythrocyte deformability via the phospholipid
membranes (Peuvot et al., 1995). Piracetam also has an effect particularly on the
phospholipids bilayers of the membranes, and it surrounds the polar heads of the
phospholipids just below the plane of interface (Peuvot et al., 1995; Vastag et al., 1999;
Fedi et al., 2001). It specifically modifies liposomes that contain phosphatidylcholine
 Piracetam and children with learning disabilities 287

and phosphatidylethanolamine (Peuvot et al., 1995); this forms a mobile complex

between the polar heads and Piracetam. This complex then takes on a conical shape,
as numerous polar heads are solvated together and form micelles or small vesicles
(Peuvot et al., 1995). This interaction alters the physical properties of the cell
membranes and increases the fluidity of the membrane (Peuvot et al., 1995; Vastag
et al., 1999), with subsequent influence on energy metabolism. It furthermore seems
as if the drug has an affinity towards the left hemisphere, thus enabling it to specifically
have an effect on the disabilities caused by injury or malformation of this region
(Wilsher et al., 1985).
Piracetam also down-regulates adhesion molecules and this enhances cerebral
blood flow specifically in the left hemispheres of the brain. This increased blood flow
is prevalent particularly in the left transverse temporal gyrus, the left posteriod supe-
rior temporal gyrus and the triangular part of the left inferior frontal temporal gyrus.
These are language-associated areas, and the taking of Piracetam improves writing
skills, comprehension, spontaneous speech as well as semantic and syntactic structure
of speech (Kessler et al., 2000).

Uses for Piracetam. Piracetam has been used to treat ischemic stroke, schizophrenia
and progressive myoclonus epilepsy; and if administrated shortly after a stroke, it was
found that it enhances the recovery of the patient’s verbal communicating abilities as
well as comprehensive and writing skills and aids in the recovery of various language
functions (Burd et al., 1997; Kessler et al., 2000). It is specifically given to aphasic
stroke patients to aid in the recovery of communication skills (Vastag et al., 1999;
Kessler et al., 2000). Shortly after a stroke, the amount of selectin-type and immuno-
globulin-type adhesion molecules in the serum increases (Vastag et al., 1999).
Burd et al. (1997) also mentioned an increase of spontaneous activity, expressive
and impulsive speech, audio-memory and speaking memory (especially delayed
memory), tactile, acoustic and visual gnosis, and space praxis in 47 acute ischemic
stroke patients treated with Piracetam from the first day of the disease on the back-
ground of basic therapy. The authors also observed more pronounced positive
dynamics of functions of the damaged hemisphere in patients with localization of
ischemic focus in the left hemisphere. Meanwhile restoration was slower when the
ischemic focus was localized in the right hemisphere. Furthermore, it was found that
restoration of high mental functions occurred faster during Piracetam treatment as
compared with basic therapy only.
Kondakor et al. (1999) also mentioned that an increased cooperativity of brain
functional processes was noted, after studying resting electroencephalogram results
from 12 healthy young volunteers after intake of a single dose of Piracetam.

Piracetam and its uses in learning disabilities

Due to the positive effect of Piracetam on language-associated areas where pathology
was involved, the question arose whether the same drug could be given to children
288 R. Britz et al.

with language-associated disabilities. Cerebral blood flow studies have indicated that
a left temporal component is used in both phonological and orthographic skills. Word
meaning is associated with the inferior left pariental region, while focal dysplasias are
found in the regions surrounding the sylvian fissure (the language region) in reading
disabled individuals (Beitchman & Young, 1997). Wilsher et al. (1985) used Pirace-
tam in conjunction with tutoring. They found an improved verbal learning, reading,
naming and tempo of reading in dyslexic children. Interestingly, the left temporal
lobe is involved in reading as suggested by Paulesu et al. (2001), and with brain imag-
ing studies (positron emission tomography) provided evidence indicating reduced left
temporal lobe activity among subjects experiencing reading difficulties.
Tallal et al. (1986) mentioned that Piracetam enhances specific cognitive skills.
The authors investigated the effect of a daily dose of 3300 mg in dyslexic boys aged
8–13 years, in a 12-week, double-blind, placebo-controlled study. Reading speed and
numbers of words written in a timed period were significantly enhanced in subjects
treated with Piracetam as compared with placebo. Effective reading and writing abil-
ity, taking both rate and accuracy into consideration, were also significantly improved
in the Piracetam as compared with the placebo treatment group. The medication was
well tolerated and medical examinations showed no significant adverse reactions.
Ackerman et al. (1991) also used Piracetam in two subgroups of students with
dyslexia and found a positive effect on verbal memory and the encoding and retrieval
of memory. Memory may be positively influenced, as mentioned previously, Piracetam
exerts a positive impact on both intracortical and cortico-subcortical conduction and
increases ATP content in the nerve cell, which leads to the increase of ribonucleic acid
synthesis necessary for long-duration memory and proteins essential for the production
of enzymes (Kawiak et al., 1995).

Applications to learning disabilities with specific reference to brain areas

It is evident that Piracetam increases blood flow in the left transverse temporal gyrus,
the left posterior superior temporal gyrus and the triangular part of the left inferior
frontal temporal gyrus, presumably without negative effect at cellular level. Further-
more, it appears that it generally enhances functioning of the temporal lobe complex.
Therefore the question rises of what is the functional involvement of the temporal
lobe during learning?

The temporal lobe: its areas and functional involvement. The temporal lobe comprises
all the tissue that lies below the Sylvian sulcus and anterior to the occipital cortex.
Subcortical temporal-lobe structures include the limbic cortex, the amygdala and the
hippocampal formation; therefore the temporal lobe does not have a unitary function,
and is particularly involved in all major areas of academic learning.
The temporal regions can be divided on the lateral surface into those that are audi-
tory (Brodmann’s areas 41, 42, and 22), and those that form the ventral visual stream
on the lateral temporal lobe (Brodmann’s areas 20, 21, 37, and 38), often referred to
 Piracetam and children with learning disabilities 289

as the inferotemporal cortex (or TE) (Kolb & Whishaw, 2003). The Sylvian fissure
contains tissue forming the insula, which includes the gustatory cortex as well as the
auditory association cortex. The superior temporal sulcus, which separates the supe-
rior and middle temporal gyri, is multimodal, receiving input from auditory, visual
and somatic regions, as well as from the other two-polymodal regions (frontal and
parietal) and the paralimbic cortex. The medial temporal region (limbic cortex)
includes the amygdala and adjacent cortex (uncus), the hippocampus and surround-
ing cortex (subiculum, entorhinal cortex, perithinal cortex), and the fusiform gyrus
(Kolb & Whishaw, 2003). The temporal lobes are internally connected to various
regions with afferent projections from the sensory systems, and efferent projections to
the parietal and frontal association regions, limbic system and basal ganglia. The
medial temporal cortex and amygdala are connected by the anterior commissure.
Furthermore, the temporal cortex reveals the following connections to other
regions (Kolb & Whishaw, 2003) that might, at a functional level, be directly involved
in learning disabilities:

● These include a hierarchical sensory pathway (subserving stimulus recognition)

that emanates from the primary and secondary auditory and visual areas, and ends
in the temporal pole.
● Visual projections that constitute the ventral stream of visual processing while the
auditory projections constitute a parallel ventral stream of auditory processing occur.
● Furthermore, the dorsal auditory pathway extends from the auditory areas to the
posterior parietal cortex, and it plays some role in detecting the spatial location of
auditory inputs.
● In addition, the polymodal pathway underlies stimulus categorization and consti-
tutes a series of parallel projections from the visual and auditory association areas
into the polymodal regions of the superior temporal sulcus.
● The medial temporal projection extends from the auditory and visual association
areas into the medial temporal region (limbic cortex), first to the perirhinal cortex,
then to the entorhinal cortex, and finally into the hippocampal formation or the
amygdala or both.
● The hippocampal projection is a major system, forming the perforant pathway, and
disturbances of this projection results in major hippocampal activity dysfunction
(Kolb & Whishaw, 2003).
● The medial temporal projection is crucial to long-term memory involved in learning
disabilities.
● Finally the frontal-lobe projection constitutes a series of parallel projections from
the association areas to the frontal lobe. The frontal-lobe projection is necessary
for various aspects of movement control, vision, short-term memory, and affect.

Sensory processes where the temporal lobe are involved include:

● Object recognition (a function of the ventral visual pathway in the temporal lobe).
● Categorization that is crucial to both perception and memory, and that requires
directed attention (a function of the superior temporal sulcus).
290 R. Britz et al.

● Matching visual and auditory information (i.e. cross-modal matching) (a function

of the superior temporal sulcus).
● The formation of memory and access to stored memories (i.e. long-term
memory)—a function of the entire ventral visual stream as well as the paralimbic
cortex of the medial temporal region (Kolb & Whishaw, 2003).
All of these preceding projections and sensory processes are functionally closely
involved in academic learning. Thus, on the basis of cortical anatomy, there is a close
association between the temporal cortex dysfunction and some types of learning
disabilities, which may particularly involve processing of auditory input, visual object
recognition, long-term storage of sensory input (i.e., memory), organization of
memory, and emotional involvement in sensory input and memory. These functions
are to a greater or lesser extent closely associated with reading, writing and arithmetic.

The temporal lobe, particularly the amygdala and hippocampus and the association with
stress-induced learning disabilities. Affective responses associated with the amygdala
may also play a major role in learning disabilities, particularly in case of stress-
induced learning disabilities such as impaired memory and stress-related deficits of
attention. Kolb and Whishaw (2003) believe that the association of sensory input and
emotion is crucial for learning because stimuli become associated with their positive,
negative or neutral consequences, and behaviour is modified accordingly. As the
amygdala becomes hypersensitive with prolonged stress, it follows that learning
behaviour is modified accordingly with associated impairment of sensory processing.
Based on the functional organization of the temporal lobe, it is clear that stress has a
particular negative impact on audio-sensory and visuo-sensory processing, as well as
on memory processing (Newport & Nemeroff, 2000; Bremner, 2001). Various
authors identified significant memory impairment in patients exposed to prolonged
stress (Sutker et al., 1991, 1995; Bremner et al., 1993, 1995; Uddo et al., 1993;
Yehuda et al., 1995; Gurvits et al., 1996). Pretorius and Naudé (2002) reported that
children presenting with chronic stress show lowered right hippocampal activation,
resulting in impaired memory. The hippocampus and medial prefrontal cortex play
important roles in memory and emotional regulation, and dysfunction in these areas
underlies memory deficits (Bremner, 1999). Furthermore, pre-clinical research
conducted over the past decade has shown that experimental stressors such as social
stress can result in functional and morphological changes within the hippocampus
(Squire & Zola-Morgan, 1991; Sapolsky, 1992, 1994; Miller et al., 1993; Zola-
Morgan et al., 1994; Alvarez et al., 1995; Richard et al., 1998). In addition, McEwen
(1997) noted that hippocampal shrinkage is usually accompanied by deficits in
declarative, episodic, spatial and contextual memory performance and the hippocam-
pal changes provide a neural substrate for changes in cognitive function that have
been recognized to accompany conditions of chronic stress.
Tanaka (1993) proved that the temporal lobe’s role in visual processing is not deter-
mined genetically, but is subject to experience (even in the adult), implicating that
processing can be either enhanced by means of therapy or impaired by environmental
 Piracetam and children with learning disabilities 291

experiences such as chronic stress. In addition, Tanaka (1993) reported that the infe-
rior temporal neurons not only process visual input, but also provide a mechanism for
the internal representation of the images of objects. Furthermore, the discharges of
these neurons provide the basis for imagery. It is thus expected that impaired inferior
temporal activity due to chronic stress would produce poor imagery associated with
learning disabilities.
As mentioned previously, temporal involvement is directly associated with auditory
processing, spatial location of auditory inputs, visual and auditory association, and
long-term memory, and by means of hierarchical pathways also indirectly involved in
short-term memory, visual object recognition, movement control, vision and affective
responses to stimulus inputs. Functional impairment associated with these are often
erroneously diagnosed as deficits of attention/hyperactivity and treated with certain
stimulant medications such as methylphenidate (Ritilin®). However, it is evident that
children with learning disabilities experience processing difficulties, particularly
difficulties processing language. Since Piracetam particularly increases blood flow in
the left transverse temporal gyrus, the left posterior superior temporal gyrus and the
triangular part of the left inferior frontal temporal gyrus, seemingly without negative
effect, it is suggested that language processing might be enhanced by administration
of Piracetam, avoiding the acknowledged negative side-effects of methylphenidate.
From the aforementioned literature, it can be concluded that Piracetam enhances
temporal lobe functions where pathology is involved and specifically in regions where
language and reading function occurs. Furthermore, temporal lobe dysfunction is
also involved in many different learning-associated disabilities. The latter may be
successfully managed by pharmaceutical intervention with Piracetam. However the
question arises regarding the safety of Piracetam, especially when used in young
children. Therefore the first level of testing would the effect of Piracetam on neuron
growth in cell culture. This will be achieved by evaluating the effect of increasing
concentrations of Piracetam on the L-929 permanent cell line (FDA standard cell line
used in toxicity studies) and chick embryo neural cells in primary culture.

Cell culture studies

The FDA approved the L-929 permanent cell line for testing of cytotoxicity, and
many researchers have since this cell line to evaluate many products (Pretorius &
Bester, 2000; Motsoane et al., 2001, 2003; Pretorius et al., 2001; Bester et al., 2003).
Primary neuron cultures have also been a popular research tool for decades (Potter &
DeMarse, 2001), as it allows easy access to individual neurons, thus importantly
increasing the knowledge of the nervous system biology as well as the neurotoxic
effects of different compounds (Richard et al., 1998). Embryonic developing systems
are highly sensitive to the effects of toxic agents and cell culture systems derived from
this tissue are equally sensitive to any toxic effects of a compound, and therefore in
this study primary cultures derived from eight-day-old chick embryos, namely chick
embryo neuron (CEN) cultures, are used. These two test systems were used and their
results compared. The reason why two different cell types were used is because,
292 R. Britz et al.

although the FDA method is an approved cytotoxicity method, neural cells exposed
to Piracetam may react differently to the L-929 cells.
The effects on cell viability and cell number were determined using 3-(4,5-
dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Sgouras & Duncan,
1990; Munetaka et al., 1996; Pariete et al., 2002 and the Crystal Violet assay (Parish
& Müllbatcher, 1983); Kueng et al., 1989; Munetaka et al., 1996).

Materials and methods

Cell cultures
The permanent cell line, namely mouse fibroblast (ATTC, CCL1 NCTC clone 929
strain designated L929), was obtained from Highveld Biological Co. (Johannesburg,
South Africa). Neural cells were cultured as a primary culture from eight-day-old
chick embryos. To achieve this, brain tissue is cut into small fragments, washed and
single-cell suspensions are obtained following enzymatic digestion with trypsin.
Fibroblasts are removed by selective attachment to the plastic surface of a cell culture
flask and the CEN are finally plated in polylysine-coated cell culture plates. The
primary cultures are maintained at 37°C and 5% CO2 content for 48 hours to allow
dendrite and axon development.
Neural cells and L929 cells were grown in Eagle’s Minimum Essential Medium
supplemented with 5% fetal calf serum, and 1% antibiotic solution (10,000 µ/ml peni-
cillin, 10,000 µg/ml streptomycin and 25 µg/ml amphotericin in 0.85% saline).
Cultures were maintained at 37°C and 5% CO2 and passaged using trypsin/ethytene-
diamine tetraacetic acid solution (0.5 g/L Trypsin, 0.2 g/L. ethytenediamine tetraacetic
acid and 8.8 g/L NaCI in Hanks Buffer). Cells were plated at a concentration of 3 ×
104 cells ml in 2 cm2 wells and were kept for 24 hours at 37°C and 5% CO2 before
conducting each experiment.

Medium containing Piracetam

A stock solution of 700 mM (100 mg/ml) Piracetam was prepared. Each cell culture,
either L929 or CEN, was exposed to serial dilutions of Piracetam from 0 to 70 mM
for 24 hours.

MTT assay
Fifty microliters of 0.1 mg/ml MTT (Sigma Chemical, Johannesburg, South Africa)
stock solution in Dulbecco’s phosphate-buffered saline was added to the medium in
each well. After incubation for a further 90 minutes at 37°C and 5% CO2, the medium
was removed and 0.2 ml isopropanol: 1 M HCI solution (24:1) was added to each
well to extract the formazan product. The plates were shaken for 10 min and absor-
bancy was measured at 545 nm using a spectrophotometer (EL900) plate reader. Cell
viability is expressed as the percentage of the control that is not exposed to the agent
 Piracetam and children with learning disabilities 293

tested. Cell viability is expressed as the mean (± standard deviation) of two and three
experiments for the L929 and CEN cells, respectively, with each experimental point
being the mean of four assays.

Crystal violet assay

Directly following 24 hours exposure to Piracetam the cells were fixed for 20 minutes
by adding 10 µl of 11% gluteraldehyde solution to the medium. The plates were then
washed well, dried and stained with 100 µl of 0.1% Crystal Violet solution for 30
minutes. Again the plates were washed and dried well before the dye was extracted
with 100 µl of a 10% acetic acid solution. The absorbancy of the extracted dye was
measured at 590 nm using a spectrophotometer (EL900) plate reader The cell
number is expressed as the percentage of the control that is not exposed to the agent
tested. The cell number is expressed as the mean (± standard deviation) of two and
two experiments for the L929 and CEN cells, respectively, with each experimental
point being the mean of four assays.

Statistical analysis
Data were statistically analyzed by student’s t-test. Within-group differences were
determined for the cell number and viability. p < 0.05 was considered statistically
significant.

Results and discussion

Cell culture studies
The L929 and CEN were exposed to serial dilutions of 0–70 mM Piracetam for 24
hours, after which the cell number and cell viability were determined using the MTT
and crystal Violet assays, respectively. The MTT assay is a bioassay that measures
the ability of a cell to convert MTT through the catalytical activity of mitochondrial
succinate dehydrogenase into insoluble purple formazan crystals. The latter is solu-
blized and the optical density is measured. Any toxic insult will result in cell damage
or death associated with a decreased ability to metabolize MTT. The L929 cell line
exposed to Piracetam showed a decrease in cell viability at 0.007 and 0.07 mM to
88% and 89% (within group, increase not significant with p = 0.33 for 0.007 mM
and p = 0.192 for 0.07 mM), respectively. In contrast, for the CEN an increase in
cell viability was observed at 0.007 mM and 0.07 mM of 109% and 118% (within
group, increase not significant with p = 0.654 for 0.007 mM and p = 0.18 for 0.07
mM) (Figure 1). At a high Pirametan concentration of 70 mM mg/ml, a decrease in
cell viability to 82% and 95% for the L929 (p = 0.046) and NEC (p = 0.376), respec-
tively, was observed. The effect of 70 mM Piracetam on L929 viability is significantly
different from the control indicating that at very high concentrations toxic effects are
observed.
294 R. Britz et al.

160

Neurons
140
L929
120
% Control (Cell (Viability)

100

80

60

40

20

0
0 0.007 0.07 0.7 7 70
Concentration (mM)
Figure 1. Effect of 24-hour exposure to 0–70 mM Piracetam on L929 and CEN cell viability mea-
sured using the MTT assay

The cell number was determined by staining the attached cells with Crystal Violet
Figure 1. Effect of 24-hour exposure to 0–70 mM Piracetam on L929 and CEN cell viability measured using the MTT assay.

that is specific for protein. A toxic insult will cause the detachment of cells from the
tissue culture surface with a resulting decrease in optical density following extraction
of the dye. For the L929 cells and CEN no changes in cell number was observed at
all concentrations studied (Figure 2) and for both cell types differences were not
significant.
Piracetam at all concentrations studied showed no significant effect on cell viability
Figure 2. Effect of 24-hour exposure to 0–70 mM Piracetam on L929 and CEN cell number measured using the Crystal Violet assay.

and number in the L929 cell line. In the primary cultures of neurons an increase in
cell viability was observed at concentrations of 0.007 mM and 0.07 mM. Gabryel
et al. (2002) have also reported an increase cell viability using the MTT assay follow-
ing exposure of rat astrocytes to 0.01 mM and 0.1 mM Piracetam for 24 hours.
Bielenberg et al. (1986) studied the effects of 1 mM Piracetam on selected key
enzymes of the energy metabolism of rat glial cells. After three weeks no effect was
observed on the activities of these enzymes that included several mitochondria-
associated enzymes. In both studies the authors used selected subpopulations of cells
such as astrocytes and glial cell cultures established from one-day-old rats, whereas
in this study primary cultures consisting of all cell types found in the brain including
astrocytes, glial cells and neurons was used. It is well documented that the developing
embryonic brain is highly sensitive to the toxic effects of drugs and for this reason
primary cultures were established from embryonic tissue rather than from adult
 Piracetam and children with learning disabilities 295

Neurons
120 L929

100
% Control (Cell (Viability)

80

60

40

20

0
0 0.007 0.07 0.7 7 70
Concentration (mM)
Figure 2. Effect of 24-hour exposure to 0–70 mM Piracetam on L929 and CEN cell number mea-
sured using the Crystal Violet assay

tissue. No studies could be found regarding the effect of high levels, greater than 1
mM Piracetam, on cell growth in vitro. In this investigation it was found that high
Piracetam levels of 7 mM and 70 mM had no effect on cell number or viability, indi-
cating that Piracetam is not cytotoxic to cells in vitro even at levels above that
normally administered. Piracetam is drug that is used frequently and over extended
periods, and therefore further studies should include the effect of repeated exposure
over a longer time periods on cell growth in vitro.

Conclusion
Learning disabilities constitute an escalating problem worldwide and treatment
involves efforts to remediate the underlying basic processing deficits, as well as to
improve associated cognitive skills (i.e. listening, comprehension, and memory).
Biological treatment that might functionally enhance processing might potentially
benefit thousands of children. However, parents are usually reluctant to subscribe to
biological treatment regimes, due to worries about possible side-effects and long-
term effects especially on the developing brain. Even though Piracetam has been
administered to children to improve reading ability and so on, this is not a routine
practice. This is first study first study to specifically study the possible adverse effects
of Piracetam on neuron growth and function. Findings are that even at high dosages
in vitro no effect is observed. This study does not address the effect of long-term and
repeated exposure; however, the results from the current research provide evidence
that Piracetam is safe to use at physiological concentrations. Although Piracetam
296 R. Britz et al.

should not be used in all cases of educational problems without regard of the
etiology, we believe that results from the current research provides evidence that
Piracetam is safe to use at physiological concentrations, and there might be some
cases where the use of Piracetam is warranted. However, before use further cell
culture and animal studies need to be embarked on to further investigate the effect of
this product on neural cells.

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