Phenotype changes of subchondral plate osteoblasts based on a rat model of ovariectomy-induced osteoarthritis

Osteoarthritis (OA) is prevalent in postmenopausal women. Subchondral bone in ovariectomized (OVX) rats might play a more important role in cartilage degeneration compared with other types of OA. How subchondral osteoblast changes in OVX rats is still unclear. Understanding of osteoblast changes obtained from OVX subchondral bone might be helpful to clarify pathogenesis of OVX-OA.

Female Sprague-Dawley rats were randomly divided into two groups: Sham (n=20) and OVX (n=20). Serum levels of Alkaline phosphatase (ALP) and C-telopeptide of type I collagen (CTX-I) were measured every one or two weeks. All rats were executed at week 9 post surgery. The weight of rats and the wet weight of uterus were assessed. Micro-computed Tomography (micro-CT) was used to analyze the knee microstructure, and toluidine blue staining was employed to evaluate cartilage erosion. Subchondral osteoblast proliferation ability by cell counting kit-8 assay, osteogenic genes expressions by reverse transcription polymerase chain reaction (RT-PCR), differentiation and mineralization ability by ALP staining and alizarin red staining were evaluated and compared between Sham and OVX.

Ovariectomy induced significant increases of serum ALP and CTX-I as early as at week 2. At week 9 after surgery, the body weight of OVX rats was significantly increased, and uterus weight of OVX rats was remarkably decreased. OVX rats demonstrated significant subchondral bone change and cartilage erosion compared with Sham rats. mRNA levels of early markers of osteogenic differentiation (ALP, type I collagen, Runx2) were enhanced in OVX rats, but the late marker (osteocalcin) was not significantly different. ALP activity of osteoblasts increased, but the mineralization capacity decreased in OVX rats.

Subchondral osteoblasts in OVX rats exhibited different proliferation, differentiation and mineralization abilities from normal counterparts.