Influences of age and mechanical stability on volume, microstructure, and mineralization of the fracture callus during bone healing: is osteoclast activity the key to age-related impaired healing?
Earlier studies have shown that the influence of fixation stability on bone healing diminishes with advanced age. The goal of this study was to unravel the relationship between mechanical stimulus and age on callus competence at a tissue level. Using 3D in vitro micro-computed tomography derived metrics, 2D in vivo radiography, and histology, we investigated the influences of age and varying fixation stability on callus size, geometry, microstructure, composition, remodeling, and vascularity. Compared were four groups with a 1.5-mm osteotomy gap in the femora of Sprague-Dawley rats: Young rigid (YR), Young semirigid (YSR), Old rigid (OR), Old semirigid (OSR). Hypothesis was that calcified callus microstructure and composition is impaired due to the influence of advanced age, and these individuals would show a reduced response to fixation stabilities. Semirigid fixations resulted in a larger DeltaCSA (Callus cross-sectional area) compared to rigid groups. In vitro microCT analysis at 6 weeks postmortem showed callus bridging scores in younger animals to be superior than their older counterparts (p<0.01). Younger animals showed (i) larger callus strut thickness (p<0.001), (ii) lower perforation in struts (p<0.01), and (iii) higher mineralization of callus struts (p<0.001). Callus mineralization was reduced in young animals with semirigid fracture fixation but remained unaffected in the aged group. While stability had an influence, age showed none on callus size and geometry of callus. With no differences observed in relative osteoid areas in the callus ROI, old as well as semirigid fixated animals showed a higher osteoclast count (p<0.05). Blood vessel density was reduced in animals with semirigid fixation (p<0.05). In conclusion, in vivo monitoring indicated delayed callus maturation in aged individuals. Callus bridging and callus competence (microstructure and mineralization) were impaired in individuals with an advanced age. This matched with increased bone resorption due to higher osteoclast numbers. Varying fixator configurations in older individuals did not alter the dominant effect of advanced age on callus tissue mineralization, unlike in their younger counterparts. Age-associated influences appeared independent from stability. This study illustrates the dominating role of osteoclastic activity in age-related impaired healing, while demonstrating the optimization of fixation parameters such as stiffness appeared to be less effective in influencing healing in aged individuals.
Julius Wolff Institute and Center for Musculoskeletal Surgery, Berlin-Brandenburg Center for Regenerative Therapies, Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, D-13353 Berlin, Germany.
M
M
Mehta
P
P
Strube
A
A
Peters
C
C
Perka
D
D
Hutmacher
P
P
Fratzl
G N
GN
Duda
D000375:Aging / Q000473:pathology*; D000818:Animals; D001696:Biomechanical Phenomena / Q000502:physiology; D015519:Bone Density / Q000502:physiology; D002146:Bony Callus / Q000000981:diagnostic imaging / Q000473:pathology*; D002113:Calcification, Physiologic / Q000502:physiology*; D005260:Female; D005592:Fracture Fixation; D017102:Fracture Healing / Q000502:physiology*; D050723:Fractures, Bone / Q000000981:diagnostic imaging / Q000503:physiopathology*; D009929:Organ Size; D010010:Osteoclasts / Q000473:pathology*; D010641:Phenotype; D011182:Postoperative Care; D051381:Rats; D017207:Rats, Sprague-Dawley; D055114:X-Ray Microtomography