Enhanced chondrogenesis and Wnt signaling in PTH-treated fractures.
Studies have shown that systemic PTH treatment enhanced the rate of bone repair in rodent models. However, the mechanisms through which PTH affects bone repair have not been elucidated. In these studies we show that PTH primarily enhanced the earliest stages of endochondral bone repair by increasing chondrocyte recruitment and rate of differentiation. In coordination with these cellular events, we observed an increased level of canonical Wnt-signaling in PTH-treated bones at multiple time-points across the time-course of fracture repair, supporting the conclusion that PTH responses are at least in part mediated through Wnt signaling.


BACKGROUND
Since FDA approval of PTH [PTH(1-34); Forteo] as a treatment for osteoporosis, there has been interest in its use in other musculoskeletal conditions. Fracture repair is one area in which PTH may have a significant clinical impact. Multiple animal studies have shown that systemic PTH treatment of healing fractures increased both callus volume and return of mechanical competence in models of fracture healing. Whereas the potential for PTH has been established, the mechanism(s) by which PTH produces these effects remain elusive.


METHODS
Closed femoral fractures were generated in 8-wk-old male C57Bl/6 mice followed by daily systemic injections of either saline (control) or 30 microg/kg PTH(1-34) for 14 days after fracture. Bones were harvested at days 2, 3, 5, 7, 10, 14, 21, and 28 after fracture and analyzed at the tissue level by radiography and histomorphometry and at the molecular and biochemical levels level by RNase protection assay (RPA), real-time PCR, and Western blot analysis.


RESULTS
Quantitative muCT analysis showed that PTH treatment induced a larger callus cross-sectional area, length, and total volume compared with controls. Molecular analysis of the expression of extracellular matrix genes associated with chondrogenesis and osteogenesis showed that PTH treated fractures displayed a 3-fold greater increase in chondrogenesis relative to osteogenesis over the course of the repair process. In addition, chondrocyte hypertrophy occurred earlier in the PTH-treated callus tissues. Analysis of the expression of potential mediators of PTH actions showed that PTH treatment significantly induced the expression of Wnts 4, 5a, 5b, and 10b and increased levels of unphosphorylated, nuclear localized beta-catenin protein, a central feature of canonical Wnt signaling.


CONCLUSIONS
These results showed that the PTH-mediated enhancement of fracture repair is primarily associated with an amplification of chondrocyte recruitment and maturation in the early fracture callus. Associated with these cellular effects, we observed an increase in canonical Wnt signaling supporting the conclusion that PTH effects on bone repair are mediated at least in part through the activation of Wnt-signaling pathways.
Kakar
Sanjeev
S
Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Boston University Medical Center, Boston, Massachusetts, USA.
Einhorn
Thomas A
TA
Vora
Siddharth
S
Miara
Lincoln J
LJ
Hon
Gregory
G
Wigner
Nathan A
NA
Toben
Daniel
D
Jacobsen
Kimberly A
KA
Al-Sebaei
Maisa O
MO
Song
Michael
M
Trackman
Philip C
PC
Morgan
Elise F
EF
Gerstenfeld
Louis C
LC
Barnes
George L
GL
D000818:Animals; D050071:Bone Density Conservation Agents / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D019902:Chondrocytes / Q000378:metabolism / Q000473:pathology; D020219:Chondrogenesis / Q000187:drug effects*; D004195:Disease Models, Animal; D005264:Femoral Fractures / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D017102:Fracture Healing / Q000187:drug effects*; D008297:Male; D051379:Mice; D015398:Signal Transduction / Q000187:drug effects*; D019379:Teriparatide / Q000494:pharmacology*; D051153:Wnt Proteins / Q000378:metabolism*
D050071:Bone Density Conservation Agents; D051153:Wnt Proteins; D019379:Teriparatide