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Lymphatic pump manipulation mobilizes inflammatory mediators into lymphatic circulation

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Lymph stasis can result in edema and the accumulation of particulate matter, exudates, toxins and bacteria in tissue interstitial fluid, leading to inflammation, impaired immune cell trafficking, tissue hypoxia, tissue fibrosis and a variety of diseases. Previously, we demonstrated that osteopathic lymphatic pump techniques (LPTs) significantly increased thoracic and intestinal duct lymph flow. The purpose of this study was to determine if LPT would mobilize inflammatory mediators into the lymphatic circulation. Under anesthesia, thoracic or intestinal lymph of dogs was collected at resting (pre-LPT), during four minutes of LPT, and for 10 min following LPT (post-LPT), and the lymphatic concentrations of interleukin-2 (IL-2), IL-4, IL-6, IL-10, interferon-γ, tissue necrosis factor α, monocyte chemotactic protein-1 (MCP-1), keratinocyte chemoattractant, superoxide dismutase (SOD) and nitrotyrosine (NT) were measured. LPT significantly increased MCP-1 concentrations in thoracic duct lymph. Further, LPT increased both thoracic and intestinal duct lymph flux of cytokines and chemokines as compared with their respective pre-LPT flux. In addition, LPT increased lymphatic flux of SOD and NT. Ten minutes following cessation of LPT, thoracic and intestinal lymph flux of cytokines, chemokines, NT and SOD were similar to pre-LPT, demonstrating that their flux was transient and a response to LPT. This re-distribution of inflammatory mediators during LPT may provide scientific rationale for the clinical use of LPT to enhance immunity and treat infection.
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Original Research
Lymphatic pump manipulation mobilizes inflammatory mediators
into lymphatic circulation
Artur Schander1, H Fred Downey2,3 and Lisa M Hodge1,3
1
Department of Molecular Biology and Immunology;
2
Department of Integrative Physiology;
3
Osteopathic Research Center,
University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107, USA
Corresponding author: Lisa M Hodge. Email: lisa.hodge@unthsc.edu
Abstract
Lymph stasis can result in edema and the accumulation of particulate matter, exudates, toxins and bacteria in tissue
interstitial fluid, leading to inflammation, impaired immune cell trafficking, tissue hypoxia, tissue fibrosis and a variety of
diseases. Previously, we demonstrated that osteopathic lymphatic pump techniques (LPTs) significantly increased thoracic
and intestinal duct lymph flow. The purpose of this study was to determine if LPT would mobilize inflammatory mediators
into the lymphatic circulation. Under anesthesia, thoracic or intestinal lymph of dogs was collected at resting (pre-LPT),
during four minutes of LPT, and for 10 min following LPT (post-LPT), and the lymphatic concentrations of interleukin-2
(IL-2), IL-4, IL-6, IL-10, interferon-
g
, tissue necrosis factor
a
, monocyte chemotactic protein-1 (MCP-1), keratinocyte
chemoattractant, superoxide dismutase (SOD) and nitrotyrosine (NT) were measured. LPT significantly increased MCP-1
concentrations in thoracic duct lymph. Further, LPT increased both thoracic and intestinal duct lymph flux of cytokines
and chemokines as compared with their respective pre-LPT flux. In addition, LPT increased lymphatic flux of SOD and NT.
Ten minutes following cessation of LPT, thoracic and intestinal lymph flux of cytokines, chemokines, NT and SOD
were similar to pre-LPT, demonstrating that their flux was transient and a response to LPT. This re-distribution of
inflammatory mediators during LPT may provide scientific rationale for the clinical use of LPT to enhance immunity and
treat infection.
Keywords: lymph, lymphatic pump technique, cytokines, chemokines, inflammatory mediators, mesenteric duct lymph,
thoracic duct lymph, infection, edema, immune system, reactive nitrogen species, reactive oxygen species, immunity,
osteopathic manipulative medicine
Experimental Biology and Medicine 2012; 237: 58 63. DOI: 10.1258/ebm.2011.011220
Introduction
Osteopathic physicians have developed osteopathic manip-
ulations collectively known as lymphatic pump techniques
(LPTs), which are designed to enhance lymph flow.
1,2
By
increasing lymph flow, LPT is thought to aid in the
removal of metabolic wastes, toxins, exudates and cellular
debris that accumulate in the tissue interstitial fluid
during infection or edema.
3
Clinically, LPT has been
shown to enhance vaccine specific antibodies,
4,5
and
reduce the length of hospital stay and the duration of anti-
biotic use in elderly patients with pneumonia.
6
During infection and edema, inflammatory cytokines, che-
mokines, reactive oxygen species (ROS), such as superoxide
dismutase (SOD), and reactive nitrogen species (RNS), such
as nitrotyrosine (NT) are generated. The proinflammatory cyto-
kines and chemokines, interleukin-2 (IL-2), IL-4, IL-6, IL-8,
tissue necrosis factor-
a
(TNF-
a
), interferon-
g
(IFN-
g
),
monocyte chemotactic protein-1 (MCP-1) and keratinocyte che-
moattractant (KC) induce leukocyte activation, migration and
cell-mediated immune responses to pathogens,
7,8
whereas anti-
inflammatory cytokines such as IL-10 limit inflammation by
suppressing cell-mediated immune responses.
7,8
Recent use of animal models has provided insight into the
mechanisms by which LPT affects the lymphatic and
immune systems.
2,9 – 12
Previously, we reported that LPT
enhances thoracic duct lymph (TDL) and mesenteric duct
lymph (MDL) flow and leukocyte concentrations in dogs
and rats.
2,10,11,13
The purpose of this study was to determine
if LPT would mobilize inflammatory mediators into the
lymphatic circulation.
7,8
In addition, SOD and NT were
measured. The results of this study provide support for
the clinical application of LPT to enhance function of the
immune system, and may explain, in part, a mechanism
by which LPT protects against infection and edema.
ISSN: 1535-3702
Copyright #2012 by the Society for Experimental Biology and Medicine
Experimental Biology and Medicine 2012; 237:5863
Materials and methods
Animals
This study was approved by the Institutional Animal Care
and Use Committee and was conducted in accordance
with the Guide for the Care and Use of Laboratory Animals
(NIH Publication no. 85-23, revised 1996). Twelve adult
mongrel dogs, free of clinically evident signs of disease,
were used for this study.
Surgical techniques
Dogs were anesthetized with sodium pentobarbital
(30 mg/kg, intravenously). After endotracheal intubation,
the dogs were ventilated with room air supplemented with
oxygen to maintain normal arterial blood gases. In addition,
arterial blood pressure was monitored via a femoral artery
catheter and remained within normal limits throughout the
experiment. In six dogs, the chest was opened by a thoracot-
omy in the left, fourth intercostal space. The thoracic duct
was isolated from connective tissue and ligated. Caudal to
the ligation, a PE 60 catheter (inner diameter 0.76 mm,
outer diameter 1.22 mm) was inserted into the duct and
secured with a ligature. Lymph was drained at atmospheric
pressure through a catheter whose outflow tip was posi-
tioned 8 cm below heart level to compensate for the hydrau-
lic resistance of the catheter. The outflow tip of the catheter
was maintained at this position for all experimental con-
ditions. Approximately 60 min following cannulation of the
thoracic duct, thoracic lymph was collected during 4 min
pre-LPT, during 4 min of LPT and for 10 min following cessa-
tion of LPT ( post-LPT). Lymph flow rate was computed from
the volume of lymph collected during these time intervals.
In separate experiments, mesenteric lymph was collected.
Six additional dogs were surgically prepared for experimen-
tation as described above. However, rather than opening the
chest, a midline abdominal incision was made to expose a
large mesenteric lymph duct. This duct was isolated,
ligated, and a PE 60 catheter was inserted into the duct
and secured with a ligature. The catheter was exteriorized
through the abdominal incision, which was then closed
with 2-0 silk suture. Approximately 60 min after cannula-
tion of the mesenteric lymph duct, mesenteric lymph
samples were collected, and lymph flow was measured as
described above for TDL.
Lymphatic pump technique
The anesthetized dogs were placed in a right lateral recumbent
position. To perform abdominal LPT, the operator contacted
the abdomen of the animal with the hands placed bilaterally
below the costo-diaphragmatic junction. Pressure was exerted
medially and cranially to compress the abdomen until signifi-
cant resistance was encountered, and then the pressure was
released. Abdominal compressions were administered at a
rate of approximately 1/s for a total of 4 min of LPT.
Measurements of TDL and MDL
A commercially available multiplex assay (Millipore,
Billerica, MA, USA) was used to determine the
concentrations of cytokines and chemokines in TDL and
MDL. Specifically, the cytokines IL-2, IL-4, IL-6, IL-10,
IFN-
g
and TNF-
a
, and the chemokines MCP-1 and KC
were measured. A range of standards, provided with the
multiplex assay, was used, and the assay was analyzed
using the Luminexw200 System with the xPONENT
Software Interface (Millipore). The minimum detectable
concentrations for IL-2, IL-4, IL-6, IL-8, IL-10, IFN-
g
,
TNF-
a
,MCP-1 and KC were 6.4, 28.8, 12.1, 20.3, 1.6, 4.4,
0.4, 8.6 and 1.6 pg/mL, respectively. To compute the cyto-
kine/chemokine flux in TDL and MDL, the respective con-
centration was multiplied by lymph flow during each
minute for each condition, and these values were averaged.
Thoracic lymph concentrations of SOD (Cayman
Chemicals, Ann Arbor, MI, USA) and NT (Molecular
Probes, Inc, Eugene, OR, USA) were measured using com-
mercially available kits. The SOD assay measures all three
forms of SOD by utilizing a tetrazolium salt for the detec-
tion of xanthine oxidase and hypoxanthine-derived super-
oxide radicals. One unit of SOD is defined as the amount
of enzyme necessary to cause 50% dismutation of the super-
oxide radical. The SOD minimum detectable concentration
for this assay is 0.025 U/mL. NO reacts with superoxide to
form peroxynitrite.
14
Subsequently, peroxynitrate reacts
with proteins, resulting in measurable NT. The minimum
detectable concentration for NT of this assay is 2 nmol/L
SOD and NT was measured only in TDL, since the
samples of MDL were not sufficient for both these measure-
ments and the Luminex assays. To compute SOD or NT flux
in TDL, the respective concentration was multiplied by
lymph flow during each minute for each condition, and
these values were averaged.
Statistical analysis
Data are presented as arithmetic means+standard error
(SE). Values from multiple animals at respective time
points were averaged and are shown in either tables or
plotted in figures. For statistical evaluation, data were sub-
jected to repeated measures analysis of variance or analysis
of variance followed by a Student NewmanKeuls mul-
tiple comparisons test. Analyses were performed with
GraphPad Prism version 5.0 for Windows (GraphPad
Software, San Diego, CA, USA). Differences among mean
values with at least P0.05 were considered statistically
significant.
Results
LPT increased intestinal and TDL flow
Similar to our previous reports,
10,11
LPT enhanced TDL
and MDL flow. LPT increased TDL flow from 0.90 +
0.19 mL/min during pre-LPT to 5.65+0.93 mL/min (P,
0.001) and the flow subsequently decreased to 2.07 +
0.28 mL/min during post-LPT (P,0.01). LPT also increased
MDL flow from 0.30+0.03 mL/min during pre-LPT to
2.71 +1.01 mL/min (P,0.05) and the flow subsequently
decreased to 0.32 +0.25 mL/min during post-LPT (P,0.05).
.................................. ..... ..... ..... ..... ..... ...... ..... ..... ..... ..... ..... ..... ..... ...... .............................. ...... ..
Schander et al.LPT and lymph cytokines 59
LPT increased the concentrations of MCP-1 in TDL
Concentrations of cytokines and chemokines in TDL and in
MDL are reported in Table 1. While cytokine and chemo-
kine concentrations in both TDL and MDL tended to
increase during LPT compared with pre- and post-LPT,
the only statistically significant increase detected was
MCP-1 in TDL (P,0.05). However, during LPT, differences
were detected between MDL and TDL in the concentrations
of IL-8 and MCP-1. Specifically, the concentration of IL-8
was greater during LPT in MDL (126%; P,0.05) compared
with TDL.
Of interest, the concentration of MCP-1 was greater in
MDL compared with TDL in all samples (Table 1).
Specifically, MCP-1 was greater at pre-LPT (435%; P,
0.01), during LPT (200%; P,0.01) and post-LPT (214%;
P,0.01) when compared with respective TDL MCP-1
concentrations.
LPT increased lymphatic cytokine and chemokine flux
The effect of LPT on flux of cytokines and chemokines in
TDL is shown in Figure 1. LPT significantly increased
TDL flux of IL-6 (615%; P,0.05), IL-8 (944%; P,0.001),
IL-10 (917%; P,0.001), MCP-1 (1505%; P,0.01) and KC
(788%; P,0.001) compared with pre-LPT. Furthermore,
these concentrations decreased post-LPT by 79% in IL-6
(P,0.05), 55% in IL-8 (P,0.01), 53% in IL-10 (P,0.01),
74% in MCP-1 (P,0.05) and 57% in KC (P,0.001).
TheeffectofLPTonfluxofcytokinesandchemokinesin
MDL is shown in Figure 2. LPT significantly increased the
MDL flux of IL-6 (394%; P,0.05), IL-8 (741%; P,0.001),
IL-10 (556%; P,0.05), MCP-1 (651%; P,0.01) and KC
(496%; P,0.001). As seen in TDL, the flux of cytokines
and chemokines in MDL declined after LPT. From LPT to
post-LPT, IL-6 decreased by 67% (P,0.05), IL-8 by 82%
(P,0.001), IL-10 by 86% (P,0.05), MCP-1 by 86% (P,
0.01) and KC by 83% (P,0.001). Cytokines IL-2, IL-4,
IFN-
g
and TNF-
a
were not detectable in TDL or in MDL
at any of the time points.
Figure 2 LPT increased cytokine and chemokine flux in mesenteric
duct lymph (MDL). Data are means +SE (n¼6). Greater than respective
pre-LPT and post-LPT (P,0.05). Greater than respective pre-LPT and
post-LPT values (P,0.01). Greater than respective pre-LPT and post-LPT
values (P,0.001). Repeated measures ANOVA with Student –Newman –Keuls
post-test. LPT, lymphatic pump technique; ANOVA, analysis of variance
Figure 1 LPT increased cytokine and chemokine flux in thoracic duct lymph
(TDL). Data are means +SE (n¼6). Greater than respective pre-LPT and
post-LPT (P,0.05). Greater than respective pre-LPT and post-LPT values
(P,0.01). Greater than respective pre-LPT and post-LPT values (P,
0.001). Repeated measures ANOVA with Student Newman Keuls post-test.
LPT, lymphatic pump technique; ANOVA, analysis of variance
Table 1 LPT significantly altered the concentration of MCP-1, but
did not significantly alter other cytokine, chemokine and reactive
oxygen species concentrations in lymph
Pre-LPT LPT Post-LPT
Thoracic duct lymph (TDL)
IL-6 (pg/mL) 193 +65 217 +74 158 +59
IL-8 (pg/mL) 183 +25 240 +48 217 +42
IL-10 (pg/mL) 24 +931+529+5
MCP-1 (pg/mL) 578 +135 1160 +304958 +266
KC (pg/mL) 1351 +165 1501 +247 1284 +172
SOD (U/mL) 0.135 +0.038 0.176 +0.026 0.173 +0.019
NT (mmol/L/mL) 3.93 +2.54 6.79 +1.93 2.86 +0.76
Mesenteric duct lymph (MDL)
IL-6 (pg/mL) 217 +43 241 +116 333 +115
IL-8 (pg/mL) 396 +85 543 +114
392 +54
IL-10 (pg/mL) 51 +20 72 +24 47 +17
MCP-1 (pg/mL) 3094 +749
3482 +685
3007 +473
KC (pg/mL) 2389 +554 2522 +506 2270 +526
LPT, lymphatic pump technique; MCP-1, monocyte chemotactic protein-1;
IL, interleukin; KC, keratinocyte chemoattractant; SOD, superoxide
dismutase; NT, nitrotyrosine
Data are means+SE (n¼6)
Greater than respective pre-LPT (P,0.05)
Different from respective TDL value (P,0.05)
Different from respective TDL value (P,0.01)
Repeated measures analysis of variance with Student Newman Keuls
post-test
.................................. ..... ..... ..... ..... ..... ...... ..... ..... ..... ..... ..... ..... ..... ...... .............................. ...... ..
60 Experimental Biology and Medicine Volume 237 January 2012
LPT increased the flux of ROS and RNS in TDL
The effect of LPT on the flux of SOD in TDL is shown in
Figure 3 and the corresponding effect on NT is shown in
Figure 4. Although LPT did not significantly increase the
concentrations of SOD and NT in TDL (Table 1), LPT
increased SOD flux 367% in TDL from 0.15 +0.07 U/min
pre-LPT to 0.7 +0.1 U/min during LPT (P,0.01).
Post-LPT, SOD flux decreased 64% to 0.25 +0.08 U/min
(P,0.01; Figure 3). LPT increased NT flux in TDL, 373%
from 5.8 +mmol/L/min pre-LPT to 27.4 +10.9 mmol/L/min
during LPT (P,0.05). Post-LPT, NT flux decreased 84% to
4.4 +1.6 mmol/L/min (P,0.05; Figure 4).
IL-6 flux was greater in TDL than in MDL during LPT
Flux of cytokines and chemokines in TDL and in MDL
during LPT is compared in Figure 5. During LPT,
IL-6 flux in TDL increased 318% more than the flux in
MDL (P,0.01).
Discussion
This study is the first to report the effects of LPT on the con-
centration and flux of inflammatory mediators in the lym-
phatic system. LPT did not significantly increase cytokine,
chemokine, ROS or RNS concentrations in lymph, with
the exception of MCP-1; however, LPT increased lymphatic
flow, which significantly increased the flux of these inflam-
matory mediators from tissue to blood via the lymphatic
system. Specifically, LPT increased the flux of IL-6, IL-8,
IL-10, MCP-1 and KC in thoracic and mesenteric lymph.
While we did not measure ROS or RNS in MDL, LPT signifi-
cantly increased SOD and NT flux in TDL. Collectively,
these results suggest that by increasing lymph flow, LPT
enhances the mobilization of inflammatory mediators into
the lymphatic circulation for transport to the blood
circulation.
Cytokines, chemokines, ROS and RNS are generated
during the innate immune response to pathogens. During
infection, the cytokines IL-6, IL-8, MCP-1 and KC induce
inflammation by recruiting and activating leukocytes,
while IL-10 regulates the inflammatory response.
7,8,15 – 17
During acute inflammation, inflammatory cytokines
Figure 3 LPT increased SOD flux in thoracic duct lymph. Data are means+
SE (n¼6). Greater than respective pre-LPT and post-LPT values (P,0.01).
Repeated measures ANOVA with Student– Newman Keuls post-test. LPT,
lymphatic pump technique; ANOVA, analysis of variance; SOD, superoxide
dismutase
Figure 5 LPT created a difference in measurable IL-6 in TDL versus MDL.
Data are means +SE (n¼6). Greater than respective MDL value (P,
0.01). ANOVA with Student– Newman Keuls post-test. LPT, lymphatic pump
technique; ANOVA, analysis of variance; IL-6, interleukin-6; MDL, mesenteric
duct lymph; TDL, thoracic duct lymph
Figure 4 LPT increased NT flux in thoracic duct lymph. Data are means +SE
(n¼6). Greater than respective pre-LPT and post-LPT values (P,0.05).
Repeated measures ANOVA with Student– Newman Keuls post-test. LPT,
lymphatic pump technique; ANOVA, analysis of variance; NT, nitrotyrosine
.................................. ..... ..... ..... ..... ..... ...... ..... ..... ..... ..... ..... ..... ..... ...... .............................. ...... ..
Schander et al.LPT and lymph cytokines 61
stimulate the formation of edema by accumulating in the
interstitial fluid, which initially lowers the interstitial fluid
pressure, setting the stage for the influx of proteins and
plasma fluid.
18,19
Therefore, LPT may suppress edema by
mobilizing inflammatory mediators out of interstitial fluid
into the lymphatic circulation, as well as directly increasing
lymph flow and removing excessive interstitial fluid.
2,12
LPT is used to treat infection,
4 – 6,20,21
but the mechanisms
by which LPT protects against infectious diseases are
unclear. LPT may enhance protection against infection by
increasing mesenteric-derived inflammatory mediators in
circulation, enabling the re-distribution of these mediators
to other tissues. In support of this notion, lymph has been
shown to re-distribute mesenteric-derived cytokines and
chemokines to distant organs.
22 – 25
Furthermore, it has
been shown in vitro that mesenteric lymph can activate neu-
trophils and increase endothelial cell permeability.
26
It is not
surprising, that LPT would enhance this re-distribution and
potentially enhance immune function.
Previously, we reported that LPT releases leukocytes from
mesenteric lymph nodes into TDL and enhances leukocyte
flux in MDL and TDL.
10
Following exposure to microorgan-
isms, phagocytes, such as macrophages and neutrophils,
release ROS and RNS which are bactericidal.
7
Thus, by
enhancing the lymphatic flux of leukocytes, cytokines, che-
mokines, ROS and RNS, LPT may facilitate cell-mediated
clearance of infection.
It has been hypothesized that following tissue injury,
lymph flow quickly increases and provides the earliest
signal in the lymphatic system to induce the inflammatory
response.
27
It has been documented that lymphedema
impairs immune cell trafficking and increases susceptibility
to infection.
28
Recently, transmural flow across lymphatic
endothelia was shown to regulate cell and fluid transport
functions of lymphatic endothelium.
29
Specifically, trans-
mural flow increased chemokine ligand secretion, influ-
enced dendritic cell migration into lymphatic vessels,
increased vessel permeability and upregulated cell adhesion
molecules on lymphatic vessels.
29
The resulting increase in
shear stress induces endothelial nitric oxide expression in
human lymphatic endothelial cells;
30
so elevated lymph
flow causes release of endogenous nitric oxide from lym-
phatic endothelial cells.
31,32
Therefore, in addition to releas-
ing leukocytes into lymphatic circulation, by enhancing
lymph flow and NT release into lymph, LPT may signal
the lymphatic system to increase immune cell trafficking.
We also compared the lymphatic cytokine and chemokine
composition between thoracic and mesenteric lymph. The
thoracic duct is a large vessel and transports lymph
drained from abdominal visceral organs (mainly the liver
and intestines), skin and skeletal muscle.
7,8,33
We found
that the concentrations of cytokines and chemokines were
higher in MDL (Table 1), which is consistent with the
prior report that most of the lymph and protein in the thor-
acic duct is derived from the mesenteric lymph.
34
This result
suggests that compared with mesenteric lymph, lymph
derived from the liver and other tissues contains low con-
centrations of inflammatory mediators, and thus dilutes
mesenteric-derived cytokines in TDL. It is important to
note that these were healthy animals; therefore, during
infection or inflammation, the concentrations of inflamma-
tory mediators in TDL and MDL may vary.
In conclusion, we have demonstrated that LPT transiently
increased the flux of chemokines, cytokines and reactive
oxygen and nitrogen species in lymph. These findings are
consistent with our previous reports, which demonstrated
that LPT transiently increases thoracic and mesenteric
lymph flow and leukocyte concentrations. This study was
performed in healthy animals, and the effect of LPT on
the lymphatic release of leukocytes and inflammatory
mediators may be intensified or altered during infection.
Our studies support the hypothesis that LPT may enhance
immune response by enhancing the release of leukocytes
and inflammatory mediators into lymphatic circulation.
Author contributions: AS performed the surgery, animal
instrumentation, statistical analysis and data interpretation,
provided the LPT and prepared the manuscript. HFD par-
ticipated in the study design, data interpretation and prep-
aration of the manuscript. LMH designed and provided
the oversight for the study. In addition, she reviewed and
interpreted the data and participated in the preparation of
the manuscript.
ACKNOWLEDGEMENTS
This study was funded by grants from the National
Institutes of Health, grants R01 AT004361 (LMH) and U19
AT002023 (HFD). The authors thank the Osteopathic
Heritage Foundation for their continued support of the
Basic Science Research Chair (LMH). The authors would
also like to thank Arthur Williams Jr and Linda Howard
for assistance in the animal surgery, and Jamie Huff and
Xin Zhang for help in the preparation of enzyme-linked
immunosorbent assays and multiplex assays.
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(Received June 20, 2011, Accepted September 10, 2011)
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Schander et al.LPT and lymph cytokines 63
... Most studies concerning the effects of OMT on inflammatory biomarkers are nonclinical. 14,[68][69][70][71] There are several studies on the effects of OMT on various conditions with inflammation-related pathologies. [72][73][74] Many of the data on OMT-induced changes in disease states lack clear biomarkers and specific endpoints. ...
Article
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Chronic inflammatory diseases (CIDs) are debilitating and potentially lethal illnesses that affect a large proportion of the global population. Osteopathic manipulative treatment (OMT) is a manual therapy technique developed and performed by osteopathic physicians that facilitates the body's innate healing processes. Therefore, OMT may prove a beneficial anti‐inflammatory modality useful in the management and treatment of CIDs. This work aims to objectively evaluate the therapeutic benefits of OMT in patients with various CIDs. In this review, a structured literature search was performed. The included studies involving asthma, chronic obstructive pulmonary disease, irritable bowel syndrome, ankylosing spondylitis, and peripheral arterial disease were selected for this work. Various OMT modalities, including lymphatic, still, counterstain, and muscle energy techniques, were utilized. Control treatments included sham techniques, routine care, or no treatment. OMT utilization led to variable patient outcomes in individuals with pathologies linked to CID.
... Other animal studies implementing LPT show increased levels of inflammatory cytokines and increased leukocyte count of all leukocyte populations, including neutrophils, monocytes, CD4+ T-cells, CD8+ T-cells, IgG, and IgA in the thoracic duct lymph that can be repeated multiple times with similar results [26][27][28]. LPTs have also been shown to recruit leukocytes from the gut-associated lymphoid tissue (GALT), which produces 60 % of immunoglobulins and where 70-80 % of plasma cells in the human body are typically located [29]. LPT has enhanced antibiotic delivery in rats infected with Streptococcus pneumoniae by diminishing bacterial load [30]. ...
Article
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Context Anecdotal evidence suggested that osteopathic manipulative treatment (OMT) may have imparted survivability to patients in osteopathic hospitals during the 1918 influenza pandemic. In addition, previous OMT research publications throughout the past century have shown evidence of increased lymphatic movement, resulting in improved immunologic function qualitatively and quantitatively. Objectives The following is a description of a proposed protocol to evaluate OMT effects on antibody generation in the peripheral circulation in response to a vaccine and its possible use in the augmentation of various vaccines. This protocol will serve as a template for OMT vaccination studies, and by adhering to the gold standard of randomized controlled trials (RCTs), future studies utilizing this outline may contribute to the much-needed advancement of the scientific literature in this field. Methods This manuscript intends to describe a protocol that will demonstrate increased antibody titers to a vaccine through OMT utilized in previous historical studies. Confirmation data will follow this manuscript validating the protocol. Study participants will be divided into groups with and without OMT with lymphatic pumps. Each group will receive the corresponding vaccine and have antibody titers measured against the specific vaccine pathogen drawn at determined intervals. Results These results will be statistically evaluated. Our demonstration of a rational scientific OMT vaccine antibody augmentation will serve as the standard for such investigation that will be reported in the future. These vaccines could include COVID-19 mRNA, influenza, shingles, rabies, and various others. The antibody response to vaccines is the resulting conclusion of its administration. Osteopathic manipulative medicine (OMM) lymphatic pumps have, in the past through anecdotal reports and smaller pilot studies, shown effectiveness on peripheral immune augmentation to vaccines. Conclusions This described protocol will be the template for more extensive scientific studies supporting osteopathic medicine’s benefit on vaccine response. The initial vaccine studies will include the COVID-19 mRNA, influenza, shingles, and rabies vaccines.
... Ранее проведенные исследования продемонстрировали снижение мышечного тонуса и улучшение кровоснабжения мышц под влиянием остеопатической коррекции [26][27][28][29], уменьшение уровня провоспалительных цитокинов и периферической сенситизации [30][31][32]. Клинические исследования продемонстрировали результативность остеопатической коррекции по сравнению с плацебо (имитация лечения) у пациентов с ГБН: у пациентов, получавших остеопатическое лечение, отмечено значительное уменьшение частоты приступов головной боли (p<0,05) и снижение использования лекарств (p<0,05) [18]. ...
Article
Introduction. Tension-type headache (TTH) is known to be the most common type of headache in all age groups. The guidelines of the European Federation of Neurological Societies, the Italian Guidelines for Primary Headaches and the Italian Consensus Conference on Pain in Neurorehabilitation report that non-pharmacological therapies are valid adjunctive treatments for TTH. Previous studies have shown that the use of general osteopathic treatment in patients with TTH is accompanied by a significant decrease in the severity of pain syndrome and asthenic condition. We did not find any scientific publications devoted to the objectification of the results of osteopathic correction in TTH using magnetic resonance imaging (MRI). The aim of the study was to objectify the results of osteopathic correction by assessing changes in the liquor dynamics of the posterior cranial fossa in patients with tension-type headache. Materials and methods . The study was conducted from December 2020 to December 2021 at the clinic of the Center for New Medical Technologies, Novosibirsk. There were under the observation 10 patients with an established diagnosis of TTH aged from 18 to 55 years, 4 men, and 6 women. All patients before the start of treatment and after the course completion were assessed for their osteopathic status and underwent high-field MRI 3T of the brain with the calculation of the posterior cranial fossa restriction index (CFRI). CFRI reflects the state of liquorodynamics at the level of the skull base and shows the level of freedom in the relationship between fluid spaces and brain tissues. Study participants received a course of osteopathic correction, which included 3–4 procedures with an interval of 5–7 days. The observed patients did not receive any other therapy during the study period. Results. The examined patients were most characterized by regional biomechanical disorders (RBD): head (9); neck, structural component (5); thoracic, visceral component (5); dura mater region (9). In terms of severity, mild RBD prevailed (1 point). After treatment, patients have a decrease of the detection frequency of major regional somatic dysfunctions (SD). Statistically significant differences (p<0,05) were obtained in the SD incidence of head region; neck region, structural component; thoracic, visceral component; dura mater region. A statistically significant (p<0,05) increase in the mean CFRI from 30,22±0,63 to 31,78±0,73 % was found after the treatment. Conclusion. The results of the high-field MRI with the study of CFRI allow to quantitatively assess the changes of the cerebrospinal fluid dynamics in patients with tension-type headache, and it can be used as an objective criterion for the osteopathic correction results and the therapy clinical effectiveness. The study should be continued with a more representative sample.
... • нормализация/уменьшение пальпаторно и визуально определяемого мышечного тонуса [16,17]; • уменьшение мышечного тонуса и улучшение кровоснабжения мышц по данным вибрационной вискоэластометрии [18,19]; • увеличение объема движений в суставах конечностей [20,21]; • увеличение объема движений в спине и шее [22,23]; • увеличение ширины открывания рта [24]; • изменение плотности коллагеновых волокон и их ориентации в структуре матрикса, уменьшение количества поперечных сшивок в них, повышение гидратации матрикса [25,26]; • изменение качественного и количественного состава внеклеточного матрикса вследствие механически индуцированных изменений синтетической активности фибробластов [7,27]; • уменьшение уровня провоспалительных цитокинов и уменьшение периферической сенситизации [28][29][30][31]; • снижение активности симпатического отдела и повышение активности парасимпатического отдела вегетативной нервной системы, восстановление вегетативного равновесия по данным анализа вариабельности сердечного ритма [12,[32][33][34]; • повышение уровня окситоцина в плазме [35]; • снижение уровня психологического стресса, уровня кортизола в крови [36]; • повышение уровня β-эндорфинов, серотонина и эндогенных каннабиноидов [37,38]; • улучшение внешнего дыхания, увеличение жизненной ёмкости легких (ЖЁЛ) [39]; • нормализация венозного давления (исходно повышенное снижалось, исходно пониженное повышалось) [40]; • активация венозного возврата к сердцу за счет увеличения подвижности грудной клетки и присасывающего действия диафрагмы [41]; • нормализация венозного оттока от головы [16,42]; • улучшение кровотока по позвоночным артериям, уменьшение асимметрии кровотока по данным УЗДГ [17,[43][44][45]; • улучшение микроциркуляции за счет выхода вазоактивных веществ из клеток соединительной ткани (оксида азота, простагландинов, гистамина и др.) [46]; • улучшение лимфообразования и лимфотока [47,48]; • выход лейкоцитов из депо [49]; • уменьшение количества внеклеточной жидкости по данным биоимпедансометрии [50]; • уменьшение вязкости тканей по данным вибрационной вискоэластометрии [51]. Таким образом, эффекты ОК могут быть по степени выраженности локальными (в виде изменения коллоидного состояния, степени гидратации и структуры ткани, улучшения подвижности сустава, изменения локальной температуры), сегментарными (в виде рефлекторных изменений) или региональными (в виде улучшения кровоснабжения и лимфотока) и глобальными (в виде гормональных эффектов, изменения функционирования ЦНС). ...
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The article describes the main objects of osteopathic influence in the body and the caused effects. The effects of osteopathic correction can occur at different times after the session and can be recorded using various clinical and instrumental methods. This should be taken into account when prescribing a re-examination of the patient to confirm the treatment results. A brief review of randomized controlled trials proving the efficacy of osteopathic correction in various diseases is also presented.
... Because of a potential toxic accumulation in IBS, the author deemed it essential to transport these waste products with lymphatic drainage techniques to improve lymphatic drainage, immunologic parameters and inflammatory markers (Huff et al., 2010;Schander, Fred Downey and Hodge, 2012;DiFrancisco-Donoghue et al., 2022). It could be considered worth trying especially in IBS-D and M situations. ...
Experiment Findings
Irritable bowel syndrome (IBS) is the most prevalent of the functional gastrointestinal disorders (FGID). It is linked with significant morbidity, biomedical and psychological problems. This illness also carries a financial burden with itself, causes impaired work related activities (lost work time, reduced productivity while at work) and a worse Quality Of Life (QOL). Osteopathy is not advised in a primary setting. For this reason a small pilot study is done to look at the possibilities.
Article
The review discusses the pathogenetic mechanisms of primary osteoarthritis (OA) formation. The recommendations of the Association of Rheumatologists of Russia on the main principles of rehabilitation for OA are presented, including drug and non-drug methods of pain syndrome correction, improvement of motor activity and quality of life of patients. Particular attention is paid to the mechanisms of osteopathic correction for OA, which are aimed at restoring the impaired biomechanics of the patient’s body, eliminating persistent muscle hypertonicity, peripheral sensitization, and aimed at regulating antinociceptive mechanisms. An important advantage of manual treatment is the possibility of its use before, during or after other types of treatment and the ability to enhance their therapeutic potential.
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Osteopathy in Russia has now formed as an independent direction of scientific knowledge and has all three characteristic levels – general philosophical, general scientific and specific scientific, as well as its own methodology. The following had been done in osteopathy as in a scientific direction of medicine: 1) its own conceptual apparatus was created; 2) a methodology had been developed that allows the use of evidence-based medicine approaches and mathematical processing of the results obtained; 3) scientific knowledge has a systematic, ordered nature; 4) the object, subject and content of osteopathy as a scientific direction were determined; 5) experimental and clinical evidence of the effectiveness of osteopathic treatment for various diseases and health disorders had been obtained. When conducting clinical studies in osteopathy, a wide range of methods for examining patients is used to obtain reliable information about the condition of organs and tissues, as well as about the body as a whole. Still it is necessary to develop the scientific component of osteopathy more actively, conduct multicenter clinical research to study clinical effectiveness, develop methodological and organizational foundations for providing osteopathic care to various groups of the population with somatic dysfunctions at all stages (prevention, diagnosis, treatment and medical rehabilitation) in order of preservation of human health, prevention of common non-communicable diseases, medical rehabilitation of patients after serious illnesses.
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The aim is to present for osteopaths the modern ideas about somatic dysfunctions as potentially reversible structural and functional disorders in the human body, and about its diagnostics and the correction possibilities. A specifi c subject of osteopathic infl uence is a group of palpable phenomena, which are called somatic dysfunctions. Somatic dysfunctions are included in the International Classifi cation of Diseases, Injuries and Conditions Affecting Health, 10th Revision (ICD-10). Somatic dysfunction (SD) is a potentially reversible structural and functional disorder in tissues and organs, manifested by palpation-determined limitations of various types of movements and mobility. Reversibility is one of the main characteristics of SD, associated with the ability to obtain the effect of changing/eliminating the identifi ed disorders in response to various methods of osteopathic correction. Impaired mobility, that is, SD, can have several components that can be combined with each other and have different degrees of severity — biomechanical, hydrodynamic (rhythmogenic) and neurodynamic. SD can manifest itself at the global, regional and local levels, and can have an acute or chronic character. The leading role in the pathogenesis of SD formation belongs to the connective tissue. Based on anamnestic data, physical examination, as well as using the algorithm of palpation diagnostic techniques, osteopaths determine the relative position of the body structures and their symmetry, as well as the qualitative state of the tissues. In addition to the generally accepted formulation of the diagnosis, an osteopathic conclusion includes the indication of biomechanical, rhythmogenic and neurodynamic disorders at the global, regional and local levels, as well as the dominant SD, the correction of which will be the logical ultimate goal of the osteopathic session. In accordance with the current regulatory framework, the osteopathic physician at the appointment fi lls out the form «Primary examination by an osteopathic physician» or the form «Examination by an osteopathic physician (observation in dynamics)». These medical documents are an insert in the Registration Form № 025/u, approved by the order of the Ministry of Health of Russia dated December 15, 2014 № 834n. The restoration of mobility is the goal of osteopathic treatment techniques applying and leads to the normalization of the functional state of tissues. The practice of osteopathy is to release the elements of the musculoskeletal system, internal organs, to restore the proper functioning of all body systems, including the nervous, circulatory and lymphatic systems. In the absence of contraindications (absolute or relative), the treatment regimen is determined individually in accordance with the issued osteopathic conclusion, including the defi ning of the number, nature (type) of techniques and the sequence of their use in a given session. The effectiveness of osteopathic correction of SD has been proven for various diseases and conditions, a list of which is also presented in the Recommendations. Conclusion . The implementation of the Clinical Recommendations can contribute to the timely diagnosis and improve the quality of medical care for patients with SD.
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Liver regeneration stimulated by a loss of liver mass leads to hepatocyte and nonparenchymal cell proliferation and rapid restoration of liver parenchyma. Mice with targeted disruption of the interleukin-6 (IL-6) gene had impaired liver regeneration characterized by liver necrosis and failure. There was a blunted DNA synthetic response in hepatocytes of these mice but not in nonparenchymal liver cells. Furthermore, there were discrete G1 phase (prereplicative stage in the cell cycle) abnormalities including absence of STAT3 (signal transducer and activator of transcription protein 3) activation and depressed AP-1, Myc, and cyclin D1 expression. Treatment of IL-6-deficient mice with a single preoperative dose of IL-6 returned STAT3 binding, gene expression, and hepatocyte proliferation to near normal and prevented liver damage, establishing that IL-6 is a critical component of the regenerative response.
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In contrast to the established role of blood vessel remodeling in inflammation, the biologic function of the lymphatic vasculature in acute inflammation has remained less explored. We studied 2 established models of acute cutaneous inflammation, namely, oxazolone-induced delayed-type hypersensitivity reactions and ultraviolet B irradiation, in keratin 14-vascular endothelial growth factor (VEGF)-C and keratin 14-VEGF-D transgenic mice. These mice have an expanded network of cutaneous lymphatic vessels. Transgenic delivery of the lymphangiogenic factors VEGF-C and the VEGFR-3 specific ligand mouse VEGF-D significantly limited acute skin inflammation in both experimental models, with a strong reduction of dermal edema. Expression of VEGFR-3 by lymphatic endothelium was strongly down-regulated at the mRNA and protein level in acutely inflamed skin, and no VEGFR-3 expression was detectable on inflamed blood vessels and dermal macrophages. There was no major change of the inflammatory cell infiltrate or the composition of the inflammatory cytokine milieu in the inflamed skin of VEGF-C or VEGF-D transgenic mice. However, the increased network of lymphatic vessels in these mice significantly enhanced lymphatic drainage from the ear skin. These results provide evidence that specific lymphatic vessel activation limits acute skin inflammation via promotion of lymph flow from the skin and reduction of edema formation.
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Lymphatic pump techniques (LPT) are used by osteopathic practitioners for the treatment of edema and infection; however, the mechanisms by which LPT enhances the lymphatic and immune systems are poorly understood. To measure the effect of LPT on the rat, the cisterna chyli (CC) of 10 rats were cannulated and lymph was collected during 4 min of 1) pre-LPT baseline, 2) 4 min LPT, and 3) 10 min post-LPT recovery. LPT increased significantly (p < 0.05) lymph flow from a baseline of 24 ± 5 μl/min to 89 ± 30 μl/min. The baseline CC lymphocyte flux was 0.65 ± 0.21 × 10⁶ lymphocytes/min, and LPT increased CC lymphocyte flux to 6.10 ± 0.99 × 10⁶ lymphocytes/min (p < 0.01). LPT had no preferential effect on any lymphocyte population, since total lymphocytes, CD4+ T cells, CD8+ T cells, and B cell numbers were similarly increased. To determine if LPT mobilized gut-associated lymphocytes into the CC lymph, gut-associated lymphocytes in the CC lymph were identified by staining CC lymphocytes for the gut homing receptor integrin α4β7. LPT significantly increased (p < 0.01) the flux of α4β7 positive CC lymphocytes from a baseline of 0.70 ± 0.03 × 10⁵ lymphocytes/min to 6.50 ± 0.10 × 10⁵ lymphocytes/min during LPT. Finally, lymphocyte flux during recovery was similar to baseline, indicating the effects of LPT are transient. Collectively, these results suggest that LPT may enhance immune surveillance by increasing the numbers of lymphocytes released in to lymphatic circulation, especially from the gut associated lymphoid tissue. The rat provides a useful model to further investigate the effect of LPT on the lymphatic and immune systems.
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Osteopathic lymphatic pump treatments (LPT) are used to treat edema, but their direct effects on lymph flow have not been studied. In the current study, we examined the effects of LPT on lymph flow in the thoracic duct of instrumented conscious dogs in the presence of edema produced by constriction of the inferior vena cava (IVC). Six dogs were surgically instrumented with an ultrasonic flow transducer on the thoracic lymph duct and catheters in the descending thoracic aorta and in IVC. After postoperative recovery, lymph flow and hemodynamic variables were measured 1) pre-LPT, 2) during 4 min LPT, 3) post-LPT, in the absence and presence of edema produced by IVC constriction. This constriction increased abdominal girth from 60 +/-2.6 to 75 +/- 2.9 cm. Before IVC constriction, LPT increased lymph flow (P < 0.05) from 1.9 +/- 0.2 ml/min to a maximum of 4.7 +/-1.2 ml/min, whereas after IVC constriction, LPT increased lymph flow (P < 0.05) from 7.9 +/-2.2 to a maximum of 11.7 +/-2.2 ml/min. The incremental lymph flow mobilized by 4 min of LPT (ie, the flow that exceeded 4 min of baseline flow), was 10.6 ml after IVC constriction. This incremental flow was not significantly greater than that measured before IVC constriction. Edema caused by IVC constriction markedly increased lymph flow in the thoracic duct. LPT increased thoracic duct lymph flow before and after IVC constriction. The lymph flow mobilized by 4 min of LPT in presence of edema was not significantly greater than that mobilized prior to edema.
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Lymphatic pump techniques (LPT) are used clinically by osteopathic practitioners for the treatment of edema and infection; however, the mechanisms by which LPT enhances lymphatic circulation and provides protection during infection are not understood. Rhythmic compressions on the abdomen during LPT compress the abdominal area, including the gut-associated lymphoid tissues (GALT), which may facilitate the release of leukocytes from these tissues into the lymphatic circulation. This study is the first to document LPT-induced mobilization of leukocytes from the GALT into the lymphatic circulation. Catheters were inserted into either the thoracic or mesenteric lymph ducts of dogs. To determine if LPT enhanced the release of leukocytes from the mesenteric lymph nodes (MLN) into lymph, the MLN were fluorescently labeled in situ. Lymph was collected during 4 min pre-LPT, 4 min LPT, and 10 min following cessation of LPT. LPT significantly increased lymph flow and leukocytes in both mesenteric and thoracic duct lymph. LPT had no preferential effect on any specific leukocyte population, since neutrophil, monocyte, CD4+ T cell, CD8+ T cell, IgG+B cell, and IgA+B cell numbers were similarly increased. In addition, LPT significantly increased the mobilization of leukocytes from the MLN into lymph. Lymph flow and leukocyte counts fell following LPT treatment, indicating that the effects of LPT are transient. LPT mobilizes leukocytes from GALT, and these leukocytes are transported by the lymphatic circulation. This enhanced release of leukocytes from GALT may provide scientific rationale for the clinical use of LPT to improve immune function.
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Background Morbidity and even mortality correlate closely with major injury that causes a systemic inflammatory response. Cytokines and bioactive molecules present at the inflammatory site induce this response and regulate neutrophil proinflammatory responses. The CXC chemokines, important for neutrophil recruitment and activation, include interleukin 8 (IL-8), granulocyte chemotactic protein 2 (GCP-2), and epithelial cell-derived neutrophil attractant 78 (ENA-78). They induce neutrophil responses via 2 cell-surface receptors, CXCR-1 and CXCR-2. All 3 chemokines bind CXCR-2 with high affinity. Only IL-8 and GCP-2 bind CXCR-1 with high affinity.Hypothesis The CXC chemokines regulate neutrophil responses differently.Methods Pretreatment of neutrophils from healthy volunteers with IL-8, GCP-2, or ENA-78; measured IL-8–induced migration; and tumor necrosis factor α (TNF-α)–induced peroxide production.Results Flow cytometry and radioligand binding data indicate that IL-8, GCP-2, and ENA-78 equivalently reduced CXCR-1 and CXCR-2 cell surface expression by 34% to 54%. All treatments decreased affinity of both receptors 1.5- to 2-fold. However, only IL-8 pretreatment inhibited chemotaxis to 10-nmol/L IL-8 (mean ± SE inhibition, 62% ± 6%). Although IL-8 and GCP-2, but not ENA-78, suppressed TNF-α–induced oxidant production (mean ± SE inhibition, 42% ± 8% and 40% ± 23%, respectively), only GCP-2 inhibited the oxidative response to complement fragment C5a, and to the bacterial cell wall peptide N-formyl-methionyl-leucyl-phenylalanine.Conclusions The CXC chemokines regulate neutrophil proinflammatory functions differently. A thorough understanding of mechanisms for modulating neutrophil responses in inflammation will aid the development of interventions that reduce morbidity and mortality associated with severe trauma and sepsis.
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The lymphatic vascular system is essential for lipid absorption, fluid homeostasis, and immune surveillance. Until recently, lymphatic vessel dysfunction had been associated with symptomatic pathologic conditions such as lymphedema. Work in the last few years had led to a better understanding of the functional roles of this vascular system in health and disease. Furthermore, recent work has also unraveled additional functional roles of the lymphatic vasculature in fat metabolism, obesity, inflammation, and the regulation of salt storage in hypertension. In this review, we summarize the functional roles of the lymphatic vasculature in health and disease.