doi: 10.1152/japplphysiol.01133.

2006
103:629-636, 2007. First published 3 May 2007; J Appl Physiol
Cassandra E. Morris and Thomas C. Skalak
enlargement resulting from surgical intervention
Chronic static magnetic field exposure alters microvessel
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Chronic static magnetic eld exposure alters microvessel enlargement
resulting from surgical intervention
Cassandra E. Morris and Thomas C. Skalak
Department of Biomedical Engineering, University of Virginia Health Sciences Center, Charlottesville, Virginia
Submitted 6 October 2006; accepted in nal form 28 April 2007
Morris CE, Skalak TC. Chronic static magnetic eld exposure
alters microvessel enlargement resulting from surgical intervention.
J Appl Physiol 103: 629636, 2007. First published May 3, 2007;
doi:10.1152/japplphysiol.01133.2006.Magnetic eld therapy has
recently become a widely used complementary/alternative medicine
for the treatment of vascular, as well as other musculoskeletal pathol-
ogies, including soft tissue injuries. Recent studies in our laboratory
and others have suggested that acute static magnetic eld (SMF)
exposure can have a modulatory inuence on the microvasculature,
acting to normalize vascular function; however, the effect of chronic
SMF exposure has not been investigated. This study aimed to mea-
sure, for the rst time, the adaptive microvascular response to a
chronic 7-day continuous magnetic eld exposure. Murine dorsal
skinfold chambers were applied on day 0, and neodymium static
magnets (or size and weight-matched shams) were afxed to the
chambers at day 0, where they remained until day 7. Separate analysis
of arteriolar and venular diameters revealed that chronic SMF appli-
cation signicantly abrogated the luminal diameter expansion ob-
served in sham-treated networks. Magnet-treated venular diameters
were signicantly reduced at day 4 and day 7 (34.3 and 54.4%,
respectively) compared with sham-treated vessels. Arteriolar diame-
ters were also signicantly reduced by magnet treatment at day 7
(50%), but not signicantly at day 4 (31.6%), although the same trend
was evident. Venular functional length density was also signicantly
reduced (60%) by chronic eld application. These results suggest that
chronic SMF exposure can alter the adaptive microvascular remodel-
ing response to mechanical injury, thus supporting the further study of
chronic application of SMFs for the treatment of vascular pathologies
involving the dysregulation of microvascular structure.
microvascular remodeling
MAGNETIC FIELD THERAPY HAS recently become a widely used
complementary or alternative medicine for the treatment of
vascular, as well as other musculoskeletal pathologies, includ-
ing soft tissue injuries. Ongoing research to investigate the
specic effects of magnetic elds in the areas of wound
healing, inammation, and the microcirculation increasingly
support the therapeutic efcacy of pulsed electromagnetic
elds (PEMF) (2, 3, 10, 14, 27, 30, 31, 40), but evidence
concerning the efcacy of static magnetic elds (SMF) is less
established. This study aimed to investigate, for the rst time,
the adaptive microvascular response to chronic magnetic eld
exposure.
Recent studies in our laboratory and others have suggested
that acute SMF exposure can have a modulatory inuence on
the microvasculature, acting to normalize vascular function.
Specically, acute SMF application can modulate microvascu-
lar tone (2123), blood pressure (25), and blood ow (47).
These results may wholly, or in part, account for the observed
benecial effects of SMF application on wound healing, in-
ammation (15, 44), collateral vessel development (6), and
decreased tumor angiogenesis (8, 42), but a direct relationship
has not been established. Variations in magnetic eld intensity,
site of application, and duration of exposure make these studies
difcult to compare, although the reported positive therapeutic
results warrant careful investigation of the vascular effects.
Studies addressing the potential therapeutic effects of
chronic exposure to SMFs are limited, as most research has
been aimed at studying the adverse effects of chronic exposure
to extremely low-frequency and radio-frequency electromag-
netic elds, similar to those emitted by power lines and cellular
telephones (9). Only a few detailed in vivo studies utilizing
sustained chronic SMF exposure have been completed to date,
but have shown measurable effects. A direct vascular effect
was observed by Xu et al. (46) as a long-lasting vasodilation of
a single arteriole after 13 wk of an 180-mT SMF exposure.
Additionally, Okano et al. reported a suppressed and delayed
onset of hypertension in spontaneously hypertensive rats ex-
posed to a 5-mT gradient eld for 212 wk (24, 26) that is
possibly related to the measured decrease in angiotensin II
(ANG II) and aldosterone levels. While these studies suggest
chronic SMF exposure may directly or indirectly inuence the
microvasculature, no assessment of the effects of a localized
magnetic eld on microvascular structural remodeling has been
completed to date.
Chronic alterations in microvascular tone resulting from
hemodynamic and/or inammatory stimuli have been shown to
induce microvascular remodeling (36, 38). Based on the exist-
ing evidence that SMF application may inuence microvascu-
lar tone (21), these experiments were designed to investigate
the effect of a locally applied chronic SMF on microvascular
structural remodeling induced by surgical intervention. The
objective of this study was to characterize the effects of a
chronic, localized SMF on microvascular network adaptations.
These results will be important for elucidation of the vascular
response to chronic, localized SMF exposure and the develop-
ment of efcacious therapeutic modalities.
METHODS
Experimental protocol. All experiments were conducted in accor-
dance with an animal protocol approved by the University of Virginia
Animal Care and Use Committee. Male c57Bl/6 mice (Harlan, Indi-
anapolis, IN) weighing 2430 g were anesthetized with an intraperi-
toneal injection of ketamine (0.00156 ml/g), xylazine (0.00052 ml/g),
and sterile saline (0.00312 ml/g), and dorsal skinfold chambers were
Address for reprint requests and other correspondence: T. C. Skalak, Dept.
of Biomedical Engineering, Univ. of Virginia Health Sciences Center, P.O.
Box 800759, Health System, Charlottesville, VA 22908 (e-mail: tskalak
@virginia.edu).
The costs of publication of this article were defrayed in part by the payment
of page charges. The article must therefore be hereby marked advertisement
in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
J Appl Physiol 103: 629636, 2007.
First published May 3, 2007; doi:10.1152/japplphysiol.01133.2006.
8750-7587/07 $8.00 Copyright 2007 the American Physiological Society http://www. jap.org 629

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applied according to the procedure outlined previously (13). Briey,
the dual layer of dorsal skin was raised, and a 7-mm 7-mm square
area of one layer of the epidermis was removed from one side of the
raised skinfold, leaving the underlying dermis (containing the micro-
vasculature), supercial fascia, and panniculus carnosus (striated skin
muscle) intact. The full thickness of the skin on the contralateral side
of the chamber was not altered. Sterile saline (0.9%) was applied
during the excision of the skin to ensure that the tissue remained
hydrated. Following excision, the skinfold was then sandwiched
perpendicular to the spine between two aluminum frames, and the
remaining tissue was covered by a sterile coverslip. A neodymium
magnet (1.39 0.01 g) or a size- and weight-matched copper sham
(1.44 0.01 g) was applied directly to the chamber after chamber
application and imaging on day 0 and removed only for imaging
purposes throughout the duration of the experiment.
Data acquisition and analysis. Intravital observation was facilitated
with a Zeiss microscope (Axioskop, Carl Zeiss, Thornwood, NY)
utilizing 2.5 (0.075 numerical aperture, Zeiss Plan Neofuar) and
10 (0.25 numerical aperture, Zeiss Plan Neofuar) objectives under
transillumination. Images of the microvasculature were taken on day
0 following surgery and subsequently on days 4 and 7 (under isou-
rane anesthesia, 2.5% isourane, 97.5% oxygen at 0.4 l/min) with an
Olympus Microre digital camera and Picture Frame Software
(Optronics, Muskogee, OK) or a charge-coupled device camera
(CCD-72, DAGE MTI, Michigan City, IN) and video recorder
(S-VHS recorder; Panasonic, Osaka, Japan). Digital images from the
charge-coupled device camera were acquired via frame grabbing
sequential images from the experimental video corresponding to the
points along the vessel to be measured. Quantication of vessel
diameters and densities was completed with the aid of Image J (NIH
Image) and Scion Image version 4.0.2 (Scion, Frederick, MD) soft-
ware packages. Three diameter measurements were taken along the
length of each vessel at each time point and were averaged to
represent the vessel diameter of that particular vessel at that given
time point. All diameters were measured from the inner lumen. Vessel
density measurements were acquired by tracing all visible vessels in
the entire window at a magnication of 100 (10 objective) and
calculating the length of the combined tracings to yield a measure-
ment of total length per tissue area. To establish maximal microvas-
cular diameter, the dorsal skinfold tissue was continually suffused for
30 min before placement of the coverslip at day 0 and after removal
of the coverslip at day 7 with adenosine (10
4
M) (4) dissolved in a
bicarbonate-buffered saline drip (137.9 mM NaCl, 4.7 mM KCl, 1.2
MM MgSO
4
, 1.9 mM CaCl
2
, 23 mM NaHCO
3
), maintained at 37C
and bubbled with 5% CO
2
in N
2
to maintain a physiological pH of 7.4.
All measurements were made in a blinded fashion, with the treatment
groups revealed after all measurements had been made.
Tissue xation, sectioning, and immunohistochemistry. Immedi-
ately following dilation of the vasculature at day 7, the dilated
morphology of the tissue was preserved by perfusion xation of the
animal followed by excision of the chamber tissue. Animals were
euthanized with pentobarbital sodium (60 mg/kg ip, Abbot Laborato-
ries, North Chicago, IL), and the entire vasculature was thoroughly
perfused with heparinized saline followed by 4% paraformeldehyde
(Sigma) in PBS solution. The paraformeldehyde solution was also
dripped on the tissue to preserve the in vivo orientation of the tissue.
Following 40 min of xation, the chamber tissue was carefully
excised and placed in a sucrose wash solution composed of 20%
sucrose and 0.02% azide in PBS at room temperature for 1 h. The
wash solution was then replaced with a gelatin gel solution composed
of 20% sucrose, 7.5% gelatin, and 0.02% azide, and placed in a 37C
water bath overnight. The tissues were then aligned vertically in
sectioning dishes along the axis of the main feeder vessels and snap
frozen in liquid nitrogen. Ten-micrometer-thick cross sections were
taken at 100-m intervals throughout the tissue and placed on gelatin-
coated slides. These sections were washed with PBS and then incu-
bated in 1:100 Alexa Fluor 488-conjugated BSI-Lectin (Molecular
Probes) and 1:200 CY3-conjugated mouse monoclonal anti-smooth
muscle -actin (SMA) (Sigma) in a staining solution (PBS, 0.1%
saponin, and 2% bovine serum albumin) at 4C overnight. Sections
were then washed in PBS and 0.1% saponin solution every 10 min for
30 min and nally mounted in a 50% PBS and glycerol solution. The
sections were analyzed with a scanning confocal microscope (Nikon
D-Eclipse-C1) and EZ.C1.22 software. Eight to twelve SMA-positive
vessels were randomly selected from each tissue, and the inner and
outer perimeter of the actin-positive area of each vessel was traced.
The area encompassed by each perimeter was calculated, and the
difference between the two was calculated to represent the medial
wall area. All measurements were made in a blinded fashion, with the
treatment groups revealed after all measurements had been made.
Magnetic eld distribution. The strength and uniformity of the
magnets used in these experiments was established by utilizing a
magnetic calibration system that generates a three-dimensional map of
the magnetic ux density (21). In summary, the system collects
three-dimensional ux density measurements by moving a gaussmeter
probe (model STD61-0202-15, Ultra Thin Transverse Probe; F. W.
Bell, Orlando, FL, linear accuracy of 0.5%) through a specied
three-dimensional volume via a computer, controller (VP9000 Series,
Velmex, Bloomeld, NY), and three coupled motorized stages (BiS-
lide Assemblies, Velmex), one corresponding to each axis. The user
species the resolution of measurement, and the computer system
(Labview version 6.0, National Instruments, Austin, TX) records the
acquired ux density measurements, as measured by the gaussmeter
(model 6010 Hall-effect gaussmeter, F. W. Bell, accuracy of 0.25%).
Spatial representation of the magnetic ux density is displayed graph-
ically, utilizing Matlab software (The MathWorks, Natick, MA),
allowing for interpretation of the ux penetration and strength at the
target tissue site. The magnet utilized in these experiments (www.
painreliver.com) was scanned with a 2-mm resolution over a 2-cm
2
area, 3 mm from the surface of the magnet. This separation distance
corresponds to the distance between the surface of the magnet and the
underlying microvasculature. Graphical representation of the three-
dimensional surface (B) and two-dimensional projection (A) of the
ux distribution at the tissue level is given in Fig. 1. The area
corresponding to the tissue within the window chamber itself is
demarcated by the inscribed square. It is noted that the eld the tissue
is exposed to is a gradient, with a maximum of 2060 mT/0.7 cm
or 2585 mT/cm. The origin of the gradient, however, is inherent to
the magnet itself; the tissue thickness in this case (400 m) does not
contribute signicantly to the gradient effect. Sham exposure in these
experiments was achieved by application of a size- and weight-
matched copper sham to the external surface of the chamber in lieu of
the magnet.
Statistical analysis. All diameter changes were compared with a
two-way ANOVA and post hoc comparisons with Tukeys test,
assuming a P value of 0.05. Vessel densities, medial wall area, and
comparisons of average maximally dilated day 0 and day 7 diameters
were completed with Students t-test, assuming a P value of 0.05
(Sigma-Stat Software, SPSS, Chicago, IL).
RESULTS
Seven-day SMF exposure abrogates vessel enlargement.
Qualitative analysis revealed a visually identiable difference
in the network morphology between the sham- and magnet-
treated tissues (Fig. 2). Untreated tissues were visually indis-
tinguishable from sham tissues (data not shown). Quantica-
tion of vessel diameters and comparison back to day 0 values
revealed marked venular enlargement in the sham-treated net-
works (n 12), 59.2 5.6% on day 4 and 91.2 8.3% on day
7, which was signicantly abrogated by magnet treatment (n
11), 38.9 4.0% on day 4 and 41.6 4.9% on day 7 (Fig. 3A).
Stratication of the data into diameter bins demonstrated that
630 MAGNETIC FIELD APPLICATION REDUCES VASCULAR REMODELING
J Appl Physiol VOL 103 AUGUST 2007 www.jap.org

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this signicant abrogation was evident in all but the largest
(75 m) initial diameter vessels (Fig. 3B). The number of
vessels measured at each time point and each initial vessel
diameter bin are given in Fig. 3, C and D, respectively.
Arteriolar enlargement was also observed in sham-treated
networks (n 10), 16.6 4.1% on day 4 and 40 4.2% on
day 7. Chronic magnet application (n 10) signicantly
reduced arteriolar enlargement at day 7 (20.5 5.1%), but not
at day 4 (11.35 3.6%), although the same trend was evident
(Fig. 3E). Separation into initial vessel diameter bins showed
that the most signicant reduction in arteriolar enlargement
was manifested in the smallest vessels, with initial diameters
Fig. 1. Graphical representation of the mag-
netic eld utilized for chronic application.
A: two-dimensional projection of the mag-
netic ux distribution applied to the tissue.
B: three-dimensional surface plot. Square in
A represents the window chamber area and,
therefore, the portion of the distribution to
which the tissue in the window is exposed.
Units are in Gauss, 10 G 1 mT.
Fig. 2. Photomicrographic images of the mi-
crovasculature treated with a chronic magnet
or sham application at day 0. Low magni-
cation of entire window (A and C) and high
magnication of a selected portion of the
window (B and D) of chronic sham- (A and B)
and magnet-treated (C and D) microvascular
networks is shown. Bar 200 m.
631 MAGNETIC FIELD APPLICATION REDUCES VASCULAR REMODELING
J Appl Physiol VOL 103 AUGUST 2007 www.jap.org

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1024 m (Fig. 3F). Again, the number of vessels measured
are given in Fig. 3, G and H.
Seven-day SMF exposure reduces venular functional length
density. Understanding that quantication of changes in micro-
vascular diameter limits the analysis to vessels that are visible
under the set magnication conditions at day 0 and also
provides no information about the pattern of the network,
quantication of total functional length density of sham- (n
8) and magnet-treated (n 9) window chambers was com-
pleted. Functional length density in this context was dened as
all perfused vessels (including visible capillaries) in the net-
work under anesthetic conditions at day 0 and day 7. At these
time points, the length of all vessels in the entire visible
network was traced and quantied (Fig. 4).
Chronic application of a SMF signicantly reduced venular
functional length density compared with sham-treated net-
works (33.3 5.7 vs. 83.4 20.5%, P 0.025). Arteriolar
functional length density was also reduced by chronic magnet
application, but not to a signicant degree (Fig. 4), in part due
to the large variability of the sham-treated arteriolar densities.
It is also interesting to note from this data that there was a
twofold greater increase in sham-treated venular length density
(83.4 20.5%) over arteriolar length density (36.1 30.8%),
suggesting that the increase in density is preferentially
weighted to the venular side of the vascular network. Specic
conclusions concerning the mechanistic origin of the observed
increase in density (formation of new vessels or enlargement of
existing vessels that were not originally visible at day 0) cannot
be directly ascertained from this data, but it is still noteworthy
that application of the static eld can have a signicant effect
on the resting, functional density of this network.
In an attempt to eliminate the possibility that the increase in
sham venular density was not primarily due to dilation of the
vasculature alone, dilated day 0 and day 7 (n 5) venular
density was quantied (data not shown). The percent change in
dilated, sham-treated, day 7 vs. day 0 length density (90.7
24.1%) was not signicantly different from undilated, sham-
treated density (83.4 20.5%). This lends support to the
assertion that the measurements made for Fig. 4, although not
dilated, represent all venules in the network, and we can
assume that no venular lumens are completely abolished under
normal anesthetic conditions. Therefore, our assessment of
nondilated diameters can be interpreted as not only a reduction
in enlarged venules, but also a reduction in new venule forma-
tion upon chronic magnetic eld application. Consequently,
magnet application cannot modulate only the caliber but also
the density of microvascular networks.
Fig. 3. Percent change in venular (A and B)
and arteriolar (E and F) diameters compared
with day 0 for all vessels (A and E) or
separated into bins by initial vessel diameter
(B and F). C, D, G, and H: number of vessels
measured corresponding to each bar in A, B,
E, and F, respectively. Values are means
SE. *P 0.05.
632 MAGNETIC FIELD APPLICATION REDUCES VASCULAR REMODELING
J Appl Physiol VOL 103 AUGUST 2007 www.jap.org

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Decreased structural wall remodeling is not via reduction of
medial wall area. To further assess the nature of the observed
increase in vessel diameter in sham-treated networks (an in-
crease in diameter via dilation vs. structural wall remodeling),
sham-treated networks were dilated and imaged at days 0 and
7 (n 9 and 8, respectively). Maximal dilation of the vascu-
lature and comparison at these time points allowed for a more
direct assessment of structural remodeling by forcing the net-
work to have uniform tone, and therefore any signicant
increase in maximal diameter may be attributed to a structural
remodeling of the vascular wall.
Comparison of the average day 0 maximal vessel diameters
to their average day 7 maximal diameters (Fig. 5) revealed a
signicant increase in both venular (49.7 3.2 vs. 80.2 5.0
m) and arteriolar (46.7 2.8 vs. 58.2 3.7 m) diameter by
day 7. These results suggest that the sham-treated networks had
undergone some structural remodeling in response to applica-
tion of the window chamber, and it was this remodeling that
was prevented by application of the magnetic eld.
Quantication of medial wall cross-sectional area is a com-
monly used metric to assess vascular wall structural remodel-
ing (38). Therefore, to investigate the nature of the observed
structural wall remodeling in sham vs. magnet networks, 10-
m-thick cross sections of dilated day 7 tissues were taken and
stained with an antibody to SMA to demarcate the medial
layer. Calculation of the maximally dilated medial cross-
sectional area from magnet- (n 6) and sham-treated (n 5)
tissues revealed no signicant difference for venules (633.7
80.1 vs. 777.6 126.2 m
2
, not signicant) or arterioles
(411.6 52.3 vs. 394.6 92.6 m
2
), respectively (Fig. 6).
This suggests that the remodeling that occurs in sham-treated
networks is not due to medial tissue hypertrophy, leaving the
hypothesis that the structural remodeling could be attributed to
the rearrangement of existing wall constituents, resulting in
larger, remodeled vessels with a decreased wall thickness.
DISCUSSION
The ability of the microvasculature to remodel both in
network topology and in single-vessel wall morphology in
response to chronic external stimuli, such as injury and inam-
mation, is critical for maintenance of a healthy vasculature.
These experiments aimed to investigate the hypothesis that
chronic SMF application could modulate vascular dilation and
thus structural microvascular adaptations in a murine dorsal
skinfold chamber.
The magnets used in these studies were carefully calibrated
utilizing a calibration system designed specically for this
purpose, allowing for precise determination of the local eld
strength acting at the target tissue location. The range of eld
strengths utilized was 2060 mT (Fig. 1). These values fall
into the range generally advertised for magnetic eld therapy
products (7) and are on the same order of magnitude as those
reported in other investigations into the efcacy of SMF
application. The dependency of dose, duration, and time of
application on signicant outcomes has not been thoroughly
Fig. 4. Representative photomicrographic montages of sham- (A and B) and magnet-treated (C and D) tissues at day 0 (A and C) and day 7 (B and D). E: percent
change in functional length density, day 7 vs. day 0, for 7-day sham- and magnet-treated skinfold chambers. Values are means SE. a, Arterioles; v, venules.
*P 0.025.
Fig. 5. A: average, sham-treated, maximal venular, and arteriolar vessel
diameters at day 0 and day 7. Representative photomicrographic images are
shown of maximally dilated day 0 (B) and day 7 (C) vessels in the murine
skinfold chamber. Values are means SE. **P 0.002, *P 0.015.
633 MAGNETIC FIELD APPLICATION REDUCES VASCULAR REMODELING
J Appl Physiol VOL 103 AUGUST 2007 www.jap.org

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investigated to date. Observations in our laboratory suggest,
however, that the application of the eld at the time of injury
(day 0 in this case) is important for affecting a signicant
physiological change (unpublished observations). Addition-
ally, it had been suggested in acute experiments that a tradi-
tional dose-response relationship does not apply to magnetic
eld exposure [Ref. 47, unpublished observations], rather that
there may exist an optimal therapeutic window of magnetic
eld strength. However, no such data have been collected with
respect to chronic experiments in general and microvessel
remodeling in particular.
Analysis of sham-treated tissues revealed a quantiable
increase in arteriolar and venular diameters 4 and 7 days after
window chamber placement. While the exact mechanism(s)
underlying this response is unknown, we postulate that the
surgical manipulation of the tissue required for chamber ap-
plication produced a local inammatory response, as mechan-
ical injury is a known stimulus for acute inammation (33),
resulting in chronically dilated vessels. As chronic vasodilation
and increased hemodynamic stimuli, such as wall shear stress
and circumferential stress, are known to induce microvascular
remodeling (36, 38, 43), the relative contribution of dilation vs.
structural remodeling to the observed increase in diameter was
established by comparison of maximally dilated diameters at
day 0 and day 7. As we found that the day 7 diameters were
signicantly larger than those of day 0, we conclude that a
portion of the measured increase in arteriolar and venular
diameters in sham-treated networks may be a result of struc-
tural wall remodeling. Application of the SMF immediately
after placement of the window chamber was found to signi-
cantly alter the formation of enlarged venules at day 4 and day
7, as well as enlarged arterioles at day 7. Based on the
conclusion that the sham networks undergo some degree of
structural remodeling, these results demonstrate for the rst
time that magnet application can reduce the enlargement of
microvessels in the network. We suggest that this is due to
modulation of vessel diameter, consistent with our previous
results (21), resulting in modulated blood ow and, therefore,
a decrease in the hemodynamic stimuli that induce microvessel
remodeling. While other researchers have found that acute
SMF exposure can affect blood ow (12, 18, 47), additional
experiments are needed to quantify the ow dynamics in
response to chronic SMF exposure.
Further characterization of the response revealed that SMF
application also signicantly reduced venular functional length
density, suggesting that magnet application cannot only mod-
ulate the caliber, but can also modify the density of microvas-
cular networks. Whether the modulation of diameter leads to
modication of the density remains to be determined. As
previous research has shown that elevated hemodynamic stim-
uli resulting from chronic dilation increases arterialization of
capillaries (1, 5, 11, 32), the increase in functional arteriolar
length density is expected, but a twofold increase in venular
density over the arteriolar density in sham-treated networks has
not been previously observed. This result is consistent, how-
ever, with previous observations that new vessels preferentially
originate on the venular side of the network (20). Whether the
magnet affects endothelial cell proliferation directly, resulting
in decreased vessel density, or if the decrease in density is a
result of a chronic alteration of vascular tone, is yet to be
determined.
Interestingly, this result is consistent with the nding that
application of a PEMF to murine 16/C mammary adenocarci-
noma tumors (45), 10 min a day for 12 days, resulted in a
signicant decrease in tumor volume, as well in angiogenesis,
measured as CD31-positive tumor area (4070%). Similar
results were observed in a nude mouse model infected with a
WiDr colon adenocarcinoma. Signicant tumor growth inhibi-
tion (up to 50%), signicant reduction of p53 expression
(58%), signicant increase (98%) in apoptotic index, and an
overall 40% reduction in tumor growth in treated animals
resulted from a 70-min daily whole body exposure for 4 wk to
a modulated (static with superimposed 50-Hz extremely low
frequency) magnetic eld with an average intensity of 3.59 mT
(41, 42). Furthermore, in a series of experiments using a
murine model with transplanted mammary adenocarcinomas,
Fig. 6. 20 confocal images of BSI-Lectin
(green) and smooth muscle -actin (SMA;
red; A) and 60 confocal images of SMA
alone (B) labeled cross sections of sham-
(top) and magnet-treated (bottom) day 7 tis-
sues. Bar 200 m. C: quantication of
medial wall area for sham- and magnet-
treated arterioles and venules. Values are
means SE.
634 MAGNETIC FIELD APPLICATION REDUCES VASCULAR REMODELING
J Appl Physiol VOL 103 AUGUST 2007 www.jap.org

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application of static electric elds and SMFs applied 812
h/day for 13 days was found to enhance the efcacy of the
chemotherapy agent adriamycin in inducing tumor regression
(8). In this study, vascular density was not assessed, but, taken
together with the PEMF results as well as our ndings, we can
postulate that SMF exposure may be effective in reducing new
vessel growth in conditions involving pathological vessel
growth.
To additionally evaluate the modulation of structural remod-
eling by application of the SMF, medial wall area was assessed
and compared between sham- and magnet-treated networks at
day 7. As no signicant difference in wall area for arterioles or
venules was measured, we can argue that, based on the signif-
icant decrease in arteriolar and venular luminal diameter mea-
sured after magnet application, although medial wall cross-
sectional area remains constant, medial wall thickness may
differ between sham- and magnet-treated vessels. This would
suggest that the remodeling of the vascular wall in the sham
networks could be the result of eutrophic remodeling, the
rearrangement of existing wall material into a larger lumen
with a decreased wall thickness maintaining constant medial
wall cross-sectional area. This suggests that the application of
the magnetic eld may have altered the sham-induced remod-
eling response.
Reconciliation of these ndings with existing data is dif-
cult, as there are only a few experiments that have investigated
the role of sustained chronic SMF application. Contrary to our
ndings that chronic SMF application results in a reduction in
vessel diameter, others observed a long-lasting dilation of a
single arteriole after 13 wk of SMF exposure, as measured by
microphotoplethysmography (MPPG) (46) in a rabbit ear
chamber. This observed vasodilation could be attributed to the
initial tone of the treated vessel. Our previous studies have
shown that initially vasoconstricted vessels dilate in response
to SMF exposure (21), and, therefore, we could conclude that
the vessels were initially vasoconstricted, resulting in a vaso-
dilatory response to the SMF exposure. The prolonged vaso-
dilation and the resultant change in hemodynamics in this study
may have resulted in a remodeling response; however, this is
impossible to discern utilizing MPPG techniques, as MPPG
measures changes in volume, not blood ow or velocity.
Additionally, application of a whole body 5-mT gradient
SMF to young, spontaneously hypertensive rats was shown to
suppress and delay the onset of hypertension, as measured by
mean arterial pressure (24, 26) and characterized by decreased
ANG II and aldosterone levels 6 wk after the onset of magnetic
eld treatment. As elevated ANG II levels have been found to
enhance inward microvascular remodeling due to chronic con-
striction (37, 39), we would expect that decreased levels would
delay the remodeling prevalent in hypertension. These results
suggest that whole body SMF exposure may have a benecial
role in the prevention of microvascular remodeling via an
indirect inuence on circulating factors that are involved in
tone regulation. The causal dependence with regard to the SMF
application, however, is difcult to assess due to the applica-
tion of the eld globally to the whole body. This result is
consistent with ours, if we deduce that the reduction in ANG II
levels corresponds to a delay in the inward remodeling.
The physiological inuence of therapeutic magnetic eld
application has been the focus of a range of in vivo, in vitro,
and clinical studies. These studies imply several possible
applications for magnetic eld therapy, including pain
management, edema reduction, modulation of tissue perfusion,
and apoptotic or anti-angiogenic treatment of tumors. The
mechanisms for these effects have not been proven unequivo-
cally, but are generally hypothesized to be related to the
activation of enzymatic reactions and/or alterations in ion
transport, specically Ca
2
, based on several in vitro ndings.
Application of a 120-mT eld to cultured GH3 cells resulted in
a reduced calcium current and activation rate of calcium
channels (19, 23, 25), as well as calcium-dependent changes in
acetylcholine release at the neuromuscular junction (34). These
results were proposed to be due to lipid reorganization in the
membrane following magnetic eld-induced increases in mem-
brane uidity. More recent investigations have observed de-
creased proliferation, inositol phosphate, and diacylglycerol
signaling in human skin broblasts (28) and FNC-B4 neuronal
cells (29) exposed for 1 h and 15 min, respectively, to a
200-mT SMF. These results further suggest that SMF may
result in decreased intracellular Ca
2
levels. This same group
also found that a 200-mT SMF applied for 3 h signicantly
inhibited PGE
1
and fetal calf serum-induced angiogenesis in
the chick embryo chorioallantoic membrane (35), further sug-
gesting a potential anti-angiogenic role of SMF exposure.
These ndings support the hypothesis that SMF exposure alters
Ca
2
dynamics, and this may relate to the change in vascular
tone. While these results are intriguing, the explanation as to
precisely how the magnetic eld is affecting these changes is
elusive. Several studies with proteins in solution have indicated
changes in chemical kinetics, such as phosphorylation events
(16, 17), which may be due to magnetically induced changes in
protein (e.g., ion channel) conguration; however, further re-
search is needed to elucidate the specic mechanism of action
and, therefore, design the most efcacious therapy.
ACKNOWLEDGMENTS
We thank Dr. Shigeru Ichioka for donating the backpacks utilized in these
experiments.
GRANTS
This work was supported by National Institutes of Health (National Center
for Complementary and Alternative Medicine) Grant AT-00582.
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