Thurs.

, 10/18/18 Basic Science: Making Bone Heal, PAPER #22

Noninvasive Localized Cold Therapy Enhances Angiogenesis in Bone Defect Model

Marianne Comeau-Gauthier, MD; Daniel Castano, MD; Geraldine Merle, PhD;
Justin Drager, MD, MsC; Edward Harvey, MD, MsC

Purpose: An OTA-funded pilot study investigated local cold therapy on defect repair in a
mouse model. Lowering the temperature at the defect site during the healing course resulted
in a significant increase in bone volume, despite a decrease of the osteoblast activity, VEGF
(vascular endothelial growth factor) expression, and CD34-stained cells. These results did
not identify the mechanism for bone growth. We hypothesized that cold therapy stimulates
neovascularization by reducing oxygen level at the injured bone, thereby upregulating
hypoxia and angiogenesis pathways. We aimed to identify the effect of cold therapy on
neovascularization during the early inflammation phase of bone repair.

Methods: Bilateral femoral window defects were drilled in adult wild-type male mice.
Starting postoperative day 1, 1 lower extremity was immerged daily in an ice bath for 15
PAPER ABSTRACTS

minutes (temperature at bone level = 19°C), whereas the other was used as control. Mice
were euthanized at day 7. Bone formation was assessed using micro CT. Histological
analysis was performed and stained for alkaline phosphatase (ALP), tartrate resistant acid
phosphatase (TRAP), CD34, and VEGF to identify osteogenic cells, osteoclasts, endothelial
vascular cells, and angiogenesis, respectively.

Results: As expected no gross new bone appeared in control or experimental groups. At

high magnification, ALP-stained sections revealed low bone forming activity for both cold
therapy and non-cold therapy groups. Femoral defects that received daily cold treatment
were associated with an excessive expression of TRAP (P = 0.03) positive cells whereas
very little osteoclast activity was detected in control group. CD34 activity was noticeably
reduced (P = 0.03) in the control group compared with the experimental. It was accompanied
by increased VEGF (P = 0.03) from 9.33 ± 6.45 for the control limbs to 15.62 ± 3.51 for the
experimental limbs.

Conclusion: The highest degree of neovascularization was observed in the defect/medulla

of cold-treated bone defects and was accompanied by a higher VEGF expression, indicating
that localized cold might support early vascularization through the VEGF angiogenesis
pathway. The overexpression of VEGF matched with the increase of osteoclast recruitment,
suggesting increased bone turnover. Bearing in mind that maintenance of bone oxygen
pressure is closely related to the blood flow, cold applied to the bone defect area may cause
hypoxia through vasoconstriction along with the usual disruption of the local blood flow
following injury, which in turn has been shown to promote osteogenesis, via its upregulation
of the angiogenesis pathway.

See the meeting app for complete listing of authors’ disclosure information.

82