3225

Influence of Localized Vibration Loading on Bone Targeting of

Bisphosphonate.
Rao V. L. Papineni*, Jingyi Pan, and Benjamin Geldhof
Carestream Molecular Imaging, 4 Research Dr., Woodbridge, CT, 06525, USA.
* rao.papineni1@carestreamhealth.com

Abstract
Targeting of bisphosphanate analog to specific skeletal sites is investigated by using localized
vibration loading. Breast, lung, and prostate cancer cells metastasize frequently at bone resulting
in debilitating pain, which is one of the major complications in majority of patients. The palliative
bone pain management includes the bisphosphanate therapy. Bisphosphonates apart from being a
potent inhibitor of bone resorption, also provides pain relief and improves bone health. However
these medications result in complications such as bisphosphonate-related osteonecrosis of the
jaw (BRONJ). Our continued efforts in investigating site-selective skeletal targeting of
bisphosphonates are primarily to establish modes to overcome such drug-related side-effects.
Non-invasive near-infrared (NIR) fluorescence imaging was used to monitor binding of NIR dye
conjugated bisphosphonate (osteosense) to specific skeletal sites in response to localized
vibration loading. Vibration loading, was performed on multiple sets of mice- subjecting them to
vibration frequency of 70 Hz for five min (5 times/week) duration at the right thigh region.
Osteosense and was injected intravenously after 21-day exercise regime. Both noninvasive and ex
vivo imaging show significantly increased NIR signals at the right surface of the lumbar vertebrae.
The unequivocal outcome in both the cohort studies, following the localized vibration loading
confirms the role of direct high-frequency vibration stimulation in bisphosphonate targeting. Multiple Single Control
Vibration Protocol: Multiple Single Control
Histochemical analysis of the bone and the muscle describe the mechanistic role in the targeted
binding of the bisphosphonate. Left Flank Region Right Flank - Skinned
Vibration Protocol: Multiple Single none (Control)
Fluorescence image Overlay of Fluorescence and X-ray images

Right Flank Region

Asymmetrical Vibration Loading Illumination Source:
Exposure Type:
Exposure Time:
Multi-wavelength
Standard Exposure
60.00 sec.
Overlay of Fluorescence and X-ray images

X-binning: No
Ex-Vivo: Fluorescence images of two different
Y-binning: No sets of skeletal structures– Prone position.
Can localized vibration loading enhance bisphosphonate binding? f-Stop: 2.80
FOV: 140 mm
Focal Plane: 11.0 mm
Vertical Resolution: 369 ppi
Horizontal Resolution: 369 ppi Localized Drug Targeting at Lumbar
Vibration loading, was performed on Magnification Stage: No
multiple sets of mice subjecting them to Excitation Filter 730 Vertebrae by Vibration Loading
vibration frequency of 70 Hz for five min Emission Filter 790 Near-IR Fluorescence (NIRF) and X-ray Imaging.
Illumination Correction Done: Yes
(5 times/week) duration at the right The animals were imaged with a multimodal (X-ray and
Illumination Source: X-Ray
thigh region. After the 21-day exercise Exposure Type: Standard Exposure optical) Carestream in-Vivo Multispectral FX Pro. The
regime, Osteosense was injected Exposure Time: 30.000 sec. representative imaging parameters are provided (left).
X-binning: No
intravenously prior to the day of near-IR The positioning status of the animals during imaging
Y-binning: No
fluorescence imaging. NMRI-Foxn1nu f-Stop: 2.80 are described in Blue boxes.
adult mice was used for the studies. FOV: 140 mm
Focal Plane: 9.9 mm
Near-IR Fluorescence (NIRF) and X-ray : points to the vibration response regions.
Vertical Resolution: 369 ppi
Imaging. Horizontal Resolution: 369 ppi : points to the regions of regular
The animals were imaged with a Magnification Stage: No physiological bone metabolic changes.
Illumination Correction Done: Yes
commercially available imaging system
- bench-top multimodal (X-ray and
optical) imaging apparatus- Carestream
Set I: Multiple Single Control Set II: Multiple Single Control
in-Vivo Multispectral FX Pro.
Immunohistochemistry of muscle sections - No molecular changes observed.
Gastrocnemius Left Right Left Right Left Right

Spine: Histology staining: Immunohistochemistry of the Lumbar Vertebrae (section #2 in the left
Multiple Multiple Multiple

Trap stain (tartrate-resistant cartoon) of mouse subjected to multiple exposures of vibration. No

acid phosphatase) for staining asymmetrical morphological and cellular changes observed. Single Single Single
of osteoclasts,
Immuohistochemistry:
Osteocalcin (BGLAP)- bone None None None
gamma-carboxyglutamic acid-
containing protein (BGLAP) for
ACTN 3 TNF IFN
osteoblasts, F4/80 for F4/80
antigen, a 160kD glycoprotein
expressed by murine
Conclusions Dr .Rao Papineni

macrophages and Vascular  Targeting of NIR bone binding drug to the ventral surface of the lumbar vertebrae is achieved through
endothelial growth factor
localized vibration loading.
(VEGF) for angiogenesis, were
performed on sections of whole  Histopathology studies rule out the possibility of any muscular or bone morphological changes in the
decalcified spine. localized drug targeting.
Gastrocnemius: Toulidine blue stain of mouse spine section. Segments 1, 2,
Immunohistochemistry: DAB 3, indicate the regions subjected to immunohistochemistry.  The induction of localized bisphosphonate targeting confirms the use of such methods as
+ chromogen was used with bisphoshonate therapeutic aid and overcome side-effects resulting from prolonged bisphosphonate use.
HRP with hematoxylin. BGLAP TRAP F4/80
Carestream is a trademark of Carestream Health, Inc. Carestream Molecular Imaging is a division of Carestream Health, Inc. The Osteosense 750 is a trademark of PerkinElmer.
Although the Carestream In-Vivo Multispectral Imaging System FX can be used for in vivo and in vitro molecular imaging of materials, researchers should be aware that the methods
of preparing and viewing the materials for molecular imaging may be subject to various patent rights.
"Molecular Imaging - Wisdom To See For Maladies To Flee"
Dr. Rao V. L. Papineni

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