IL-6 Reduces Spheroid Sizes of Osteophytic Cells Derived from Osteoarthritis Knee Joint via Induction of Apoptosis.

Osteophytes in osteoarthritis (OA) joints contribute to restriction of joint movement, joint pain, and OA progression, but little is known about osteophyte regulators. Examination of gene expression related to cartilage extracellular matrix, endochondral ossification, and growth factor signaling in articular cartilage and osteophytes obtained from OA knee joints showed that several genes such as COL1A1, VCAN, BGLAP, BMP8B, RUNX2, and SOST were overexpressed in osteophytes compared with articular cartilage. Ratios of mesenchymal stem/progenitor cells, which were characterized by co-expression of CD105 and CD166, were significantly higher in osteophytic cells than articular cells. A three-dimensional culture method for cartilage and osteophyte cells was developed by modification of cultures of self-assembled spheroid cell organoids (spheroids). These spheroids cultured in the media for mesenchymal stem cells containing transforming growth factor-ß3 showed characteristic morphologies and gene expression profiles of articular cartilage and osteophytes, respectively. The effects of IL-1ß, tumor necrosis factor-α, and IL-6 on the spheroids of articular and osteophytic cells were studied. To the best of our knowledge, they provide the first evidence that IL-6 suppresses the spheroid size of osteophytic cells by inducing apoptosis and reducing extracellular matrix molecules. These data show that IL-6 is the suppressor of osteophyte growth and suggest that IL-6 expression and/or activity are implicated in the regulation of osteophyte formation in pathologic joints.

Asporin and transforming growth factor-beta gene expression in osteoblasts from subchondral bone and osteophytes in osteoarthritis.

BACKGROUND: To clarify the significance of subchondral bone and osteophytes in the pathology of osteoarthritis (OA), we investigated the expression of asporin (ASPN), transforming growth factor-beta1 (TGF-beta1), TGF-beta2, TGF-beta3, and runt-related transcription factor-2 (Runx2) genes involved in bone metabolism. METHODS: Osteoblasts were isolated from 19 patients diagnosed with knee OA and from 4 patients diagnosed with femoral neck fracture. Osteoblast expression of mRNA encoding ASPN, TGF-beta1, TGF-beta2, TGF-beta3, and Runx2 was analyzed using real-time RT-PCR. RESULTS: Expression of ASPN, TGF-beta1, and TGF-beta3 mRNA in the subchondral bone and osteophytes of OA patients increased compared with that of non-OA patients. The ratio of ASPN to TGF-beta1 mRNA in patients with severe cartilage damage was higher than that in patients with mild cartilage damage. CONCLUSIONS: The increased ratio of ASPN mRNA to TGF-beta1 mRNA in patients with severe relative to mild cartilage damage indicates that increased ASPN mRNA expression was significantly associated with the severity of cartilage degeneration. This finding suggests that ASPN may regulate TGF-beta1-mediated factors in the development of OA, which may provide clues as to the underlying pathology of OA.

Puppy line, metaphyseal sclerosis, and caudolateral curvilinear and circumferential femoral head osteophytes in early detection of canine hip dysplasia.

Ventrodorsal extended hip radiographs were analyzed from Foxhounds, Irish setters, Greyhounds, and Labrador retrievers radiographed four to seven times between 8 and 110 weeks of age. Occurrence in these 91 dogs of a puppy line, an ill-defined zone of proximal femoral metaphyseal sclerosis, a femoral neck linear sclerosis, or circumferential linear femoral head osteophytosis at 15-17 weeks of age were compared with hip joint laxity, as measured by distraction index, and to later findings of caudal curvilinear femoral neck osteophytes, circumferential femoral head osteophytes, hip incongruity consistent with hip dysplasia and degenerative joint disease by 52 weeks of age. A puppy line and/or femoral metaphyseal sclerosis was common at 15-17 weeks of age for dogs at mimimal risk (Greyhounds) and high risk (Foxhounds) of developing early degenerative joint disease associated with canine hip dysplasia. Though 44% of Greyhound hips had puppy lines and 28% had femoral metaphyseal sclerosis at 15-17 weeks of age, no Greyhound had a caudolateral curvilinear osteophyte or circumferential femoral head osteophyte at 24-27 or 52 weeks of age. No significant relationship was found between occurrence of a puppy line, a circumferential femoral head osteophyte or femoral metaphyseal sclerosis at 15-17 weeks and canine hip dysplasia or degenerative joint disease incidence at 42-52 weeks. Presence of a caudolateral curvilinear osteophyte in at least one hip at 24-27 weeks was significantly related to the diagnosis of canine hip dysplasia by 42-52 weeks. When both a caudolateral curvilinear osteophyte and a circumferential femoral head osteophyte were present in a hip at 24-27 weeks, degenerative joint disease was evident in all such hips by 42-52 weeks of age.

Characterization of a mesenchymal-like stem cell population from osteophyte tissue.

Osteophytes are a distinct feature of osteoarthritis (OA). Their formation may be related to pluripotential cells in the periosteum responding to stimulus during OA. This study aimed to isolate stem cells from osteophyte tissues and to characterize their phenotype, proliferation, and differentiation potential, as well as their immunomodulatory properties. Osteophyte-derived cells were isolated from osteophyte tissue samples collected during knee replacement surgery. These cells were characterized by the expression of cell-surface antigens, differentiation potential into mesenchymal lineages, growth kinetics, and modulation of alloimmune responses. Multipotential stem cells were identified from all osteophyte samples, namely osteophyte-derived mesenchymal stem cells (oMSCs). The surface antigen expression of oMSCs was consistent with that of MSCs; they lacked the hematopoietic and common leukocyte markers (CD34, CD45) while expressing those related to adhesion (CD29, CD166, CD44) and stem cells (CD90, CD105, CD73). The proliferation capacity of oMSCs in culture was superior to that of bone marrow-derived MSCs (bMSCs), and these cells readily differentiated into tissues of the mesenchymal lineages. oMSCs also demonstrated the ability to suppress allogeneic T cell proliferation, which was associated with the expression of the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO). Our results showed that osteophyte-derived cells had similar properties to MSCs in the expression of antigen phenotype, differential potential, and suppression of alloimmune response. Furthermore, when compared to bMSCs, oMSCs maintained a higher proliferative capacity, which may offer new insights of the tissue formation and potentially an alternative source for therapeutic stem cell-based tissue regeneration.