Cathepsin K predicts femoral neck bone mineral density change in nonosteoporotic peri- and early postmenopausal women.

OBJECTIVE: Cathepsin K is a cysteine protease that plays an essential role in organic bone matrix degradation. The aim of our study was to seek correlation of serum cathepsin K levels and a change in bone mineral density (BMD) over a 3-year period in a population of healthy nonosteoporotic women. The secondary end points were the correlations of serum cathepsin K with cross-sectional BMD and with other serum bone turnover markers and age. DESIGN: In 43 healthy women aged 42 to 57 years, blood samples for determination of cathepsin K, osteocalcin, bone alkaline phosphatase, C-terminal cross-linking telopeptide of type I collagen, osteoprotegerin, and nuclear factor kappaB ligand were collected at the time of the first BMD measurement. BMD measurements were repeated after 3 years. RESULTS: We found a moderate negative correlation of serum cathepsin K levels with change in femoral neck BMD, but none with change in spinal BMD. There were no significant correlations between cross-sectional BMD of the spine or femoral neck and serum levels of cathepsin K. Serum levels of cathepsin K were not significantly correlated with any bone turnover markers studied or with age. CONCLUSIONS: Serum cathepsin K does not seem to represent a surrogate for bone turnover markers used at present, but it might be useful as a predictor of cortical bone loss.

Osteoblastogenesis and adipogenesis are higher in osteoarthritic than in osteoporotic bone tissue.

BACKGROUND AND AIMS: New data show that increased adipogenesis in bone marrow may decrease osteoblastogenesis, resulting in osteoporosis (OP). Runt-related transcription factor 2 (RUNX2) and peroxisome proliferator-activated receptor γ (PPARγ) are two main transcriptional regulators controlling osteoblastogenesis and adipogenesis from the same precursor cell in bone-the mesenchymal stem cell. Because osteoarthritis (OA) and OP present the opposing bone phenotype, our aim was to determine whether the expression of selected adipogenic genes is lower in OA compared to OP bone tissue. METHODS: Bone samples were obtained from gender-matched OP (n = 54) and OA (n = 49) patients undergoing hip arthroplasty. Osteoblastogenesis and adipogenesis were estimated by gene expression analysis of RUNX2, PPARγ2 and their downstream genes. RESULTS: In OA bone, significantly higher expression of PPARγ2 and adiponectin as well as RUNX2, osterix and osteocalcin were obtained, suggesting higher adipogenesis and osteoblastogenesis in OA than in OP. There were no differences in RUNX2/PPARγ2 and osteocalcin/adiponectin ratios between groups, suggesting similar balance of both processes. Higher perilipin 2, angiopoietin-like 4 and fatty-acid binding protein 4 mRNA levels in OP suggest activation of other transcription factors or hypoxic conditions in OP bone. CONCLUSIONS: Regulation of bone formation by RUNX2 and PPARγ2 is modified in OA compared to OP, resulting in higher osteoblastogenesis and adipogenesis in OA. Both processes are similarly balanced in OP and OA but less active in OP.

A microarray based identification of osteoporosis-related genes in primary culture of human osteoblasts.

Genetic factors influencing the pathogenesis of osteoporosis are still largely unknown. We employed genome-wide gene expression approach in order to discover novel genes involved in the pathogenesis of osteoporosis. To this end, primary cultures of osteoblasts isolated from osteoporotic and non-osteoporotic human bone tissue samples were prepared. One thousand six hundred six genes were found to be differentially expressed, indicating increased demand for protein synthesis and decreased cell proliferation rate in osteoblasts from osteoporotic tissue as compared to osteoblasts from non-osteoporotic tissue. At first, top four genes, based on the microarray data and potential role in bone metabolism, were further studied in bone tissue samples of 55 patients. PTN and COL15A1 were both downregulated in osteoporotic bone tissue (6.2- and 3.4-fold, respectively, both p<0.05), while IBSP and CXCL2 were both upregulated (5.7-fold, p<0.05, and 2.1-fold, p>0.05). Further biostatistical analysis of the microarray data by gene set enrichment analysis suggested oxidative stress may have an important part in the pathogenesis of osteoporosis. Thus, secondly, we tested it by an in vitro assay on human osteosarcoma cell line cells treated with hydrogen peroxide. After 72 h of treatment with 500 microM hydrogen peroxide, the upregulation of the same genes involved in the response to oxidative stress as on the microarrays was observed: MT1G (metallothionein 1G, 22.1-fold, p<0.05), TXNRD1 (thioredoxin reductase 1, 3.7-fold, p<0.05), AOX1 (aldehyde oxidase 1, 24.5-fold, p<0.05) and GSR (glutathione reductase, 4.7-fold, p<0.05). Our results present a novel list of genes and metabolic pathways that may be associated with the pathogenesis of osteoporosis. PTN, CXCL2, COL15A1, IBSP, AOX1, MT1G, GSR and TXNRD1 are candidate genes for further studies in the assessment of the genetic susceptibility to osteoporosis. In addition, differences in protein synthesis, cell proliferation rate and response to oxidative stress may also be involved in the pathogenesis of osteoporosis.

Expression of bone resorption genes in osteoarthritis and in osteoporosis.

Cathepsin K and MMP-9 are considered to be the most abundant proteases in osteoclasts. TRAP is a marker for osteoclasts, and there is increasing evidence of its proteolytic role in bone resorption. RANKL is a recently discovered regulator of osteoclast maturation and activity and induces expression of many genes. This study compared cathepsin K, MMP-9, TRAP, RANKL, OPG, and osteocalcin gene expression in the proximal femur of patients with osteoarthritis with that of patients with femoral neck fracture. Fifty-six patients undergoing arthroplasty because of osteoarthritis or femoral neck fracture were included in the study. Total mRNA was extracted from the bone samples obtained from the intertrochanteric region of the proximal femur. Real-time RT-PCR was used to quantify CTSK (cathepsin K), MMP-9 (matrix metalloproteinase 9), ACP5 (TRAP), TNFSF11 (RANKL), TNFRSF11B (OPG), and BGLAP (osteocalcin) mRNAs. The levels of mRNAs coding for MMP-9 and osteocalcin indicated higher expression in the osteoarthritic group (P = 0.011, P = 0.001, respectively), whereas RANKL expression and the ratio RANKL/OPG were both significantly lower in the osteoarthritic group than in the fracture group. Expression of cathepsin K, MMP-9, and TRAP relative to RANKL was significantly higher in the osteoarthritic group. Ratios of all three proteolytic enzymes relative to formation marker osteocalcin were higher in the fracture group. Gene expression of cathepsin K, MMP-9, TRAP, RANKL, OPG, and osteocalcin and the association between their mRNA levels pointed to higher bone resorption and bone formation in osteoarthritis, differences in balance between them, and differences in regulation of bone resorption in osteoarthritic and osteoporotic bone.