Sex-specific compromised bone healing in female rats might be associated with a decrease in mesenchymal stem cell quantity.
BACKGROUND
The clinically known importance of patient sex as a major risk factor for compromised bone healing is poorly reflected in animal models. Consequently, the underlying cellular mechanisms remain elusive. Because mesenchymal stem cells (MSCs) are postulated to regulate tissue regeneration and give rise to essential differentiated cell types, they may contribute to sex-specific differences in bone healing outcomes.


METHODS
We investigated sex-specific variations in bone healing and associated differences in MSC populations. A 1.5 mm osteotomy gap in the femora of 8 male and 8 female 12-month-old Sprague-Dawley rats was stabilized by an external fixator. Healing was analyzed in terms of biomechanical testing, bridging and callus size over time (radiography at 2, 4, and 6 weeks after surgery), and callus volume and geometry by microCT at final follow-up. MSCs were obtained from bone marrow samples of an age-matched group of 12 animals (6 per gender) and analyzed for numbers of colony-forming units (CFUs) and their capacity to differentiate and proliferate. The proportion of senescent cells was determined by beta-galactosidase staining.


RESULTS
Sex-specific differences were indicated by a compromised mechanical competence of the callus in females compared with males (maximum torque at failure, p=0.028). Throughout the follow-up, the cross-sectional area of callus relative to bone was reduced in females (p< or =0.01), and the bridging of callus was delayed (p(2weeks)=0.041). microCT revealed a reduced callus size (p=0.003), mineralization (p=0.003) and polar moment of inertia (p=0.003) in female animals. The female bone marrow contained significantly fewer MSCs, represented by low CFU numbers in both femora and tibiae (p(femur)=0.017, p(tibia)=0.010). Functional characteristics of male and female MSCs were similar.


CONCLUSIONS
Biomechanically compromised and radiographically delayed bone formation were distinctive in female rats. These differences were concomitant with a reduced number of MSCs, which may be causative for the suboptimal bone healing.
Strube
Patrick
P
Center for Musculoskeletal Surgery and Julius Wolff Institute Berlin, Charité University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Mehta
Manav
M
Baerenwaldt
Anne
A
Trippens
Jessica
J
Wilson
Cameron J
CJ
Ode
Andrea
A
Perka
Carsten
C
Duda
Georg N
GN
Kasper
Grit
G
D000818:Animals; D001696:Biomechanical Phenomena; D001842:Bone and Bones / Q000000981:diagnostic imaging / Q000473:pathology*; D002146:Bony Callus / Q000000981:diagnostic imaging / Q000473:pathology; D002452:Cell Count; D003114:Colony-Forming Units Assay; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012727:Sex Characteristics; D014945:Wound Healing; D055114:X-Ray Microtomography