Osteoarthritis induction leads to early and temporal subchondral plate porosity in the tibial plateau of mice: An in vivo micro CT study

Authors

Sander M. Botter, Gerjo J.V.M van Osch, Stefan Clockaerts, Jan H. Waarsing, Harrie Weinans, Johannes P.T.M. van Leeuwen

Abstract

In osteoarthritis (OA) changes occur both in cartilage and subchondral bone. The subchondral bone plate facilitates normal crosstalk between articular cartilage and trabecular subchondral bone, and adaptive changes in the plate due to OA may therefore disturb crosstalk homeostasis. To investigate these changes over time we examined the cartilage-subchondral bone interface using a combined approach of histology and in vivo micro CT. Male C57Bl/6 mice (n=8), aged 16 weeks, received intra-articular injections with collagenase in one joint to induce instability-related OA and saline into the contralateral knee joint (controls). At 2, 4, 6, 10 and 14 weeks post-injection, changes in the tibial subchondral bone plate and subchondral trabeculae were analyzed. At two weeks post-injection, collagenase injected joints had significantly more cartilage damage and osteophytosis than control joints. Osteoclast activity directly underneath the subchondral bone plate was significantly elevated (Oc.S./BS controls: 7.60± 0.81%; OA: 11.07± 0.79%), causing the plate to become thinner and creating a large increase in subchondral bone plate porosity (cumulative porosity volume controls: 0.05e-3 ± 0.04e-3 mm3; OA: 2.52e-3 ± 0.69e-3 mm3). At four weeks post-injection, the previously formed perforations disappeared, coinciding with a significant rise in osteoblast activity in the subchondral trabecular bone (bone formation rate controls: 0.30± 0.03, OA: 0.62± 0.13 μm2/μm3*day). The current study provides for the first time quantitative longitudinal data on the dynamic changes in the subchondral bone plate after OA induction. The development of plate perforations may enhance mutual interaction between subchondral trabeculae, bone marrow cells and the articular cartilage in OA.

Link to Article

http://dx.doi.org/10.1002/art.30307

Depth of subchondral perforation influences the outcome of bone marrow stimulation cartilage repair

Authors

Hongmei Chen, Caroline D. Hoemann, Jun Sun, Anik Chevrier, Marc D. McKee, Matthew S. Shive, Mark Hurtig, Michael D. Buschmann

Abstract

Subchondral drilling and microfracture are bone marrow stimulation techniques commonly used for the treatment of cartilage defects. Few studies to date have examined the technical variants which may influence the success of the cartilage repair procedures. This study compared the effect of hole depth (6 mm vs. 2 mm) and hole type (drill vs. microfracture) on chondral defect repair using a mature rabbit model. Results from quantitative histomorphometry and histological scoring showed that deeper versus shallower drilling elicited a greater fill of the cartilage defect with a more hyaline character in the repair matrix indicated by significant improvement (p = 0.021) in the aggregate measure of increased cartilage defect fill, increased glycosaminoglycan and type II collagen content and reduced type I collagen content of total soft repair tissue. Compared to microfracture at the same 2 mm depth, drilling to 2 mm produced a similar quantity and quality of cartilage repair (p = 0.120) according to the aggregate indicator described above. We conclude that the depth of bone marrow stimulation can exert important influences on cartilage repair outcomes

Link to Article

http://dx.doi.org/10.1002/jor.21386

Bone biopsy findings and correlation with clinical, radiological and biochemical parameters in children with fractures

Authors

Mervi K Mäyränpää, Inari S Tamminen, Heikki Kröger, Outi Mäkitie

Abstract

In children the diagnosis of osteoporosis is based on fracture history and DXA-derived BMD. Bone biopsy is an invasive but accurate method to study bone characteristics. In this study we evaluated bone biopsy findings and their correlation with non-invasive measures of bone health. Transiliac bone biopsy was performed on 24 consecutive children (17 boys, median age 12 years, range 6-16 years) evaluated for suspected primary osteoporosis. Biopsy findings were compared with normative data and correlated with clinical, radiological, biochemical, and densitometric findings. The patients had sustained altogether 64 non-vertebral fractures (median 2.5) from low- or moderate-energy traumas, and 14 patients (58%) had vertebral fractures. The median lumbar spine BMD Z-score was –1.2 (range -3.1 to +1.0). Hypovitaminosis D was present in 58%. Histomorphometry showed low bone volume in 7 patients and normal bone volume in 17. Bone turnover was high in 7, low in 7, and normal in 10 patients. Histomorphometric findings correlated poorly with fracture history, serum bone turnover markers and DXA findings. Vitamin D deficiency and low lumbar BMD were associated with high bone turnover in the biopsy. These findings underscore the difficulties in diagnosing pediatric osteoporosis. Bone histomorphometry gives additional information and may be useful when considering bisphosphonate treatment in children with suspected primary osteoporosis.

Link to Article

http://dx.doi.org/10.1002/jbmr.373

Bone marrow stromal cells with a combined expression of BMP-2 and VEGF-165 enhanced bone regeneration

Authors

Caiwen Xiao1,5, Huifang Zhou1,5, Guangpeng Liu2, Peng Zhang3, Yao Fu1, Ping Gu1, Hongliang Hou4, Tingting Tang4 and Xianqun Fan

Abstract

Bone graft substitutes with osteogenic factors alone often exhibit poor bone regeneration due to inadequate vascularization. Combined delivery of osteogenic and angiogenic factors from biodegradable scaffolds may enhance bone regeneration. We evaluated the effects of bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor (VEGF), combined with natural coral scaffolds, on the repair of critical-sized bone defects in rabbit orbits. In vitro expanded rabbit bone marrow stromal cells (BMSCs) were transfected with human BMP2 and VEGF165 genes. Target protein expression and osteogenic differentiation were confirmed after gene transduction. Rabbit orbital defects were treated with a coral scaffold loaded with BMP2-transduced and VEGF-transduced BMSCs, BMP2-expressing BMSCs, VEGF-expressing BMSCs, or BMSCs without gene transduction. Volume and density of regenerated bone were determined by micro-computed tomography at 4, 8, and 16 weeks after implantation. Neovascularity, new bone deposition rate, and new bone formation were measured by immunostaining, tetracycline and calcein labelling, and histomorphometric analysis at different time points. The results showed that VEGF increased blood vessel formation relative to groups without VEGF. Combined delivery of BMP2 and VEGF increased new bone deposition and formation, compared with any single factor. These findings indicate that mimicking the natural bone development process by combined BMP2 and VEGF delivery improves healing of critical-sized orbital defects in rabbits.

Link to Article

http://dx.doi.org/10.1088/1748-6041/6/1/015013

β2-Adrenergic Receptor Signaling in Osteoblasts Contributes to the Catabolic Effect of Glucocorticoids on Bone

Authors

Yun Ma, Jeffry S. Nyman, Huan Tao, Heather H. Moss, Xiangli Yang and Florent Elefteriou

Abstract

The sympathetic nervous system is a physiological regulator of bone homeostasis. Autonomic nerves are indeed present in bone, bone cells express the β2-adrenergic receptors (β2AR), and pharmacological or genetic disruption of sympathetic outflow to bone induces bone gain in rodents. These recent findings implied that conditions that affect β2AR signaling in osteoblasts and/or sympathetic drive to bone may contribute to bone diseases. In this study, we show that dexamethasone stimulates the expression of the β2AR in differentiated primary calvarial osteoblasts, as measured by an increase in Adrβ2 mRNA and β2AR protein level after short-term dexamethasone treatment. Isoproterenol-induced cAMP accumulation and the expression of the β2AR target gene Rankl were also significantly increased after dexamethasone pretreatment, indicating that dexamethasone promotes the responsiveness of differentiated osteoblasts to adrenergic stimulation. These in vitro results led to the hypothesis that glucocorticoid-induced bone loss, provoked by increased endogenous or high-dose exogenous glucocorticoids given for the treatment of inflammatory diseases, might, at least in part, be mediated by increased sensitivity of bone-forming cells to the tonic inhibitory effect of sympathetic nerves on bone formation or their stimulatory effect on bone resorption. Supporting this hypothesis, both pharmacological and genetic β2AR blockade in mice significantly reduced the bone catabolic effect of high-dose prednisolone in vivo. This study emphasizes the importance of sympathetic nerves in the regulation of bone homeostasis and indicates that this neuroskeletal signaling axis can be modulated by hormones or drugs and contribute to enhance pathological bone loss.

Link to Article

http://dx.doi.org/10.1210/en.2010-0881

Proteinase-activated Receptor-2 Gene Disruption Limits the Effect of Osteoarthritis on Cartilage in Mice: A Novel Target in Joint Degradation

Authors

Nathalie Amiable, Johanne Martel-Pelletier, Bertrand Lussier, Steeve Kwan Tat, Jean-Pierre Pelletier and Christelle Boileau

Abstract

Evidence indicates that proteinase-activated receptor (PAR)-2 participates in the degradative processes of human osteoarthritis (OA). We evaluated the in vivo effect of PAR-2 on articular lesions in a PAR-2-knockout (KO) mouse model of OA. OA was surgically induced by destabilization of the medial meniscus of the right knee in C57Bl/6 wild-type (WT) and PAR-2 KO mice. Knee swelling was measured throughout the duration of the study (8 weeks postsurgery) and histologic evaluation of cartilage was done to assess structure, cellularity, matrix staining, and remodeling in the deep zone. Morphometric analysis of subchondral bone was also performed. Data showed significant knee swelling in the operated WT mice immediately following surgery, which increased with time (8 weeks post-surgery). Knee swelling was significantly lower (p ≤ 0.0001) in PAR-2 KO mice than in WT mice at both 4 and 8 weeks postsurgery. Cartilage damage was found in both operated WT and PAR-2 KO mice; however, lesions were significantly less severe (global score; p ≤ 0.05) in the PAR-2 KO mice at 4 weeks postsurgery. Operated WT mice showed reduced subchondral bone surface and trabecular thickness with significance reached at 4 weeks (p ≤ 0.03 and p ≤ 0.05, respectively), while PAR-2 KO mice demonstrated a gradual increase in subchondral bone surface with significance reached at 8 weeks (p ≤ 0.007). We demonstrated the in vivo implication of PAR-2 in the development of experimental OA, thus confirming its involvement in OA joint structural changes and reinforcing the therapeutic potential of a PAR-2 antagonist for treatment of OA.

Link to Article

http://dx.doi.org/10.3899/jrheum.100710