HOXA10 promotes Gdf5 expression in articular chondrocytes.

Growth differentiation factor 5 (GDF5), a BMP family member, is highly expressed in the surface layer of articular cartilage. The GDF5 gene is a key risk locus for osteoarthritis and Gdf5-deficient mice show abnormal joint development, indicating that GDF5 is essential in joint development and homeostasis. In this study, we aimed to identify transcription factors involved in Gdf5 expression by performing two-step screening. We first performed microarray analyses to find transcription factors specifically and highly expressed in the superficial zone (SFZ) cells of articular cartilage, and isolated 11 transcription factors highly expressed in SFZ cells but not in costal chondrocytes. To further proceed with the identification, we generated Gdf5-HiBiT knock-in (Gdf5-HiBiT KI) mice, by which we can easily and reproducibly monitor Gdf5 expression, using CRISPR/Cas9 genome editing. Among the 11 transcription factors, Hoxa10 clearly upregulated HiBiT activity in the SFZ cells isolated from Gdf5-HiBiT KI mice. Hoxa10 overexpression increased Gdf5 expression while Hoxa10 knockdown decreased it in the SFZ cells. Moreover, ChIP and promoter assays proved the direct regulation of Gdf5 expression by HOXA10. Thus, our results indicate the important role played by HOXA10 in Gdf5 regulation and the usefulness of Gdf5-HiBiT KI mice for monitoring Gdf5 expression.

Semaphorin 4D induces articular cartilage destruction and inflammation in joints by transcriptionally reprogramming chondrocytes.

Proinflammatory cytokines play critical roles in the pathogenesis of joint diseases. Using a mass spectrometry-based cloning approach, we identified Semaphorin 4D (Sema4D) as an inflammatory cytokine that directly promoted cartilage destruction. Sema4d-deficient mice showed less cartilage destruction than wild-type mice in a model of rheumatoid arthritis. Sema4D induced a proinflammatory response in mouse articular chondrocytes characterized by the induction of proteolytic enzymes that degrade cartilage, such as matrix metalloproteinases (MMPs) and aggrecanases. The activation of Mmp13 and Mmp3 expression in articular chondrocytes by Sema4D did not depend on RhoA, a GTPase that mediates Sema4D-induced cytoskeletal rearrangements. Instead, it required NF-κB signaling and Ras-MEK-Erk1/2 signaling downstream of the receptors Plexin-B2 and c-Met and depended on the transcription factors IκBζ and C/EBPδ. Genetic and pharmacological blockade of these Sema4D signaling pathways inhibited MMP induction in chondrocytes and cartilage destruction in femoral head organ culture. Our results reveal a mechanism by which Sema4D signaling promotes cartilage destruction.

Transcriptional regulation of FRZB in chondrocytes by Osterix and Msx2.

INTRODUCTION: Osteoarthritis is a common joint disease that causes destruction of articular cartilage and severe inflammation surrounding knee and hip joints. However, to date, effective therapeutic reagents for osteoarthritis have not been developed because the underlying molecular mechanisms are complex. Recent genetic findings suggest that a Wnt antagonist, frizzled-related protein B (FRZB), is a potential therapeutic target for osteoarthritis. Therefore, this study aimed to examine the transcriptional regulation of FRZB in chondrocytes. MATERIALS AND METHODS: Frzb/FRZB expression was assessed by RT-qPCR analyses in murine articular chondrocytes and SW1353 chondrocyte cell line. Overexpression and knockdown experiments were performed using adenovirus and lentivirus, respectively. Luciferase-reporter and chromatin immunoprecipitation assays were performed for determining transcriptional regulation. Protein-protein interaction was determined by co-immunoprecipitation analysis. RESULTS: Frzb was highly expressed in cartilages, especially within articular chondrocytes. Interleukin-1α markedly reduced Frzb expression in articular chondrocytes in association with cartilage destruction and increases in ADAM metallopeptidase with thrombospondin type 1 motif (Adamts) 4 and Adamts5 expression. Bone morphogenetic protein 2 (BMP2) increased FRZB expression in SW1353 cells through Smad signaling. Osterix and msh homeobox 2 (Msx2), both of which function as downstream transcription factors of BMP2, induced FRZB expression and upregulated its promoter activity. Co-immunoprecipitation results showed a physical interaction between Osterix and Msx2. Knockdown of either Osterix or Msx2 inhibited BMP2-dependent FRZB expression. Chromatin immunoprecipitation indicated a direct association of Osterix and Msx2 with the FRZB gene promoter. CONCLUSION: These results suggest that BMP2 regulates FRZB expression through Osterix and Msx2.

Regulatory Mechanisms of Prg4 and Gdf5 Expression in Articular Cartilage and Functions in Osteoarthritis.

Owing to the rapid aging of society, the numbers of patients with joint disease continue to increase. Accordingly, a large number of patients require appropriate treatment for osteoarthritis (OA), the most frequent bone and joint disease. Thought to be caused by the degeneration and destruction of articular cartilage following persistent and excessive mechanical stimulation of the joints, OA can significantly impair patient quality of life with symptoms such as knee pain, lower limb muscle weakness, or difficulty walking. Because articular cartilage has a low self-repair ability and an extremely low proliferative capacity, healing of damaged articular cartilage has not been achieved to date. The current pharmaceutical treatment of OA is limited to the slight alleviation of symptoms (e.g., local injection of hyaluronic acid or non-steroidal anti-inflammatory drugs); hence, the development of effective drugs and regenerative therapies for OA is highly desirable. This review article summarizes findings indicating that proteoglycan 4 (Prg4)/lubricin, which is specifically expressed in the superficial zone of articular cartilage and synovium, functions in a protective manner against OA, and covers the transcriptional regulation of Prg4 in articular chondrocytes. We also focused on growth differentiation factor 5 (Gdf5), which is specifically expressed on the surface layer of articular cartilage, particularly in the developmental stage, describing its regulatory mechanisms and functions in joint formation and OA pathogenesis. Because several genetic studies in humans and mice indicate the involvement of these genes in the maintenance of articular cartilage homeostasis and the presentation of OA, molecular targeting of Prg4 and Gdf5 is expected to provide new insights into the aetiology, pathogenesis, and potential treatment of OA.

Activation and Function of NLRP3 Inflammasome in Bone and Joint-Related Diseases.

Inflammation is a pivotal response to a variety of stimuli, and inflammatory molecules such as cytokines have central roles in the pathogenesis of various diseases, including bone and joint diseases. Proinflammatory cytokines are mainly produced by immune cells and mediate inflammatory and innate immune responses. Additionally, proinflammatory cytokines accelerate bone resorption and cartilage destruction, resulting in the destruction of bone and joint tissues. Thus, proinflammatory cytokines are involved in regulating the pathogenesis of bone and joint diseases. Interleukin (IL)-1 is a representative inflammatory cytokine that strongly promotes bone and cartilage destruction, and elucidating the regulation of IL-1 will advance our understanding of the onset and progression of bone and joint diseases. IL-1 has two isoforms, IL-1α and IL-1ß. Both isoforms signal through the same IL-1 receptor type 1, but the activation mechanisms are completely different. In particular, IL-1ß is tightly regulated by protein complexes termed inflammasomes. Recent research using innovative technologies has led to a series of discoveries about inflammasomes. This review highlights the current understanding of the activation and function of the NLRP3 (NOD-like receptor family, pyrin domain-containing 3) inflammasome in bone and joint diseases.

Zfhx4 regulates endochondral ossification as the transcriptional platform of Osterix in mice.

Endochondral ossification is regulated by transcription factors that include SRY-box transcription factor 9, runt-related protein 2 (Runx2), and Osterix. However, the sequential and harmonious regulation of the multiple steps of endochondral ossification is unclear. This study identified zinc finger homeodomain 4 (Zfhx4) as a crucial transcriptional partner of Osterix. We found that Zfhx4 was highly expressed in cartilage and that Zfhx4 deficient mice had reduced expression of matrix metallopeptidase 13 and inhibited calcification of cartilage matrices. These phenotypes were very similar to impaired chondrogenesis in Osterix deficient mice. Coimmunoprecipitation and immunofluorescence indicated a physical interaction between Zfhx4 and Osterix. Notably, Zfhx4 and Osterix double mutant mice showed more severe phenotype than Zfhx4 deficient mice. Additionally, Zfhx4 interacted with Runx2 that functions upstream of Osterix. Our findings suggest that Zfhx4 coordinates the transcriptional network of Osterix and, consequently, endochondral ossification.

Smoc1 and Smoc2 regulate bone formation as downstream molecules of Runx2.

Runx2 is an essential transcription factor for bone formation. Although osteocalcin, osteopontin, and bone sialoprotein are well-known Runx2-regulated bone-specific genes, the skeletal phenotypes of knockout (KO) mice for these genes are marginal compared with those of Runx2 KO mice. These inconsistencies suggest that unknown Runx2-regulated genes play important roles in bone formation. To address this, we attempted to identify the Runx2 targets by performing RNA-sequencing and found Smoc1 and Smoc2 upregulation by Runx2. Smoc1 or Smoc2 knockdown inhibited osteoblastogenesis. Smoc1 KO mice displayed no fibula formation, while Smoc2 KO mice had mild craniofacial phenotypes. Surprisingly, Smoc1 and Smoc2 double KO (DKO) mice manifested no skull, shortened tibiae, and no fibulae. Endochondral bone formation was also impaired at the late stage in the DKO mice. Collectively, these results suggest that Smoc1 and Smoc2 function as novel targets for Runx2, and play important roles in intramembranous and endochondral bone formation.

Dmrt2 promotes transition of endochondral bone formation by linking Sox9 and Runx2.

Endochondral bone formation is fundamental for skeletal development. During this process, chondrocytes undergo multiple steps of differentiation and coordinated transition from a proliferating to a hypertrophic stage, which is critical to advance skeletal development. Here, we identified the transcription factor Dmrt2 (double-sex and mab-3 related transcription factor 2) as a Sox9-inducible gene that promotes chondrocyte hypertrophy in pre-hypertrophic chondrocytes. Epigenetic analysis further demonstrated that Sox9 regulates Dmrt2 expression through an active enhancer located 18 kb upstream of the Dmrt2 gene and that this enhancer's chromatin status is progressively activated through chondrocyte differentiation. Dmrt2-knockout mice exhibited a dwarf phenotype with delayed initiation of chondrocyte hypertrophy. Dmrt2 augmented hypertrophic chondrocyte gene expression including Ihh through physical and functional interaction with Runx2. Furthermore, Dmrt2 deficiency reduced Runx2-dependent Ihh expression. Our findings suggest that Dmrt2 is critical for sequential chondrocyte differentiation during endochondral bone formation and coordinates the transcriptional network between Sox9 and Runx2.

Molecular Mechanisms Involved in the Progression and Protection of Osteoarthritis.

OBJECTIVE: Osteoarthritis is a common disease of the joint cartilage. Since the molecular pathogenesis of osteoarthritis is not clearly understood, early diagnostic markers and effective therapeutic agents have not been developed. METHODS AND RESULTS: In recent years, there are several studies to elucidate the molecular aspects based on mouse genetics by using a stress-induced mechanical load model. Chondrocyte hypertrophy, which is usually seen in growth plate chondrocyte, is also induced in articular cartilage and involved in the onset of osteoarthritis. Additionally, signal molecules involved in inflammatory cytokine and matrix proteinase are expected to be target molecules for the fundamental treatment of early osteoarthritis. Some additional signal molecules, transcription factors and compounds have been reported to be involved in cartilage homeostasis. CONCLUSION: This review sheds light on the current status of various signal molecules for the management of osteoarthritis.

Feasibility of a self-assembling peptide hydrogel scaffold for meniscal defect: An in vivo study in a rabbit model.

The inner avascular zone of the meniscus has limited healing capacity as the area is poorly vascularized. Although peptide hydrogels have been reported to regenerate bone and cartilage, their effect on meniscus regeneration remains unknown. We tested whether the self-assembling peptide hydrogel scaffold KI24RGDS stays in the meniscal lesion and facilitates meniscal repair and regeneration in an induced rabbit meniscal defect model. Full-thickness (2.0 mm diameter) cylindrical defects were introduced into the inner avascular zones of the anterior portions of the medial menisci of rabbit knees (n = 40). Right knee defects were left empty (control group) while the left knee defects were transplanted with peptide hydrogel (KI24RGDS group). Macroscopic meniscus scores were significantly higher in the KI24RGDS group than in the control group at 2, 4, and 8 weeks after surgery. Histological examinations including quantitative and qualitative scores indicated that compared with the control group, the reparative tissue in the meniscus was significantly enhanced in the KI24RGDS group at 2, 4, 8, and 12 weeks after surgery. Immunohistochemical staining showed that the reparative tissue induced by KI24RGDS at 12 weeks postimplantation was positive for Type I and II collagen. KI24RGDS is highly biocompatible and biodegradable, with strong stiffness, and a three dimensional structure mimicking native extracellular matrix and RGDS sequences that enhance cell adhesion and proliferation. This in vivo study demonstrated that KI24RGDS remained in the meniscal lesion and facilitated the repair and regeneration in a rabbit meniscal defect model.

Histological Analysis of the Wrapping Treatment for Meniscal Horizontal Tears in Rabbits.

OBJECTIVE: To investigate meniscal regeneration and prevent cartilage degeneration using wrapping treatment for meniscal horizontal tears that have been difficult to repair in rabbits. DESIGN: Thirty knees from 15 Japanese white rabbits were divided into the horizontal (horizontal tears) or wrapping (horizontal tears with wrapping treatment) groups. Horizontal tears were created and wrapped with a sheet scaffold containing polyglycolic acid, polylactic acid, and polycaprolactone. The meniscus was stained with Safranin-O/Fast Green and evaluated with modified Pauli scores at 8, 12, and 16 weeks after implantation (n = 5). Cell morphology was determined with hematoxylin and eosin staining. Mature collagen was confirmed with Picrosirius Red staining. Furthermore, immunohistochemical analysis of inducible nitric oxide synthase (iNOS) for inflammation, Ki-67 for proliferation, and type II collagen for regeneration was performed. Medial femoral cartilage was stained with Safranin-O/Fast Green and evaluated with the Osteoarthritis Research Society International score at 8 and 16 weeks. RESULTS: The wrapping group had significantly better regeneration than the horizontal group, especially at 16 weeks (P < 0.05). Wrapping treatment induced fibrochondrocyte-like cells at 16 weeks. After wrapping treatment, iNOS was overexpressed at 8 weeks, Ki-67 at 8 and 12 weeks, and type II collagen at 16 weeks. Cartilage degeneration in the wrapping group did not progress significantly compared with that in the horizontal group at 16 weeks (P < 0.05). CONCLUSIONS: Wrapping treatment for meniscal horizontal tears induced meniscal regeneration as the sheet scaffold might induce intrinsic and extrinsic repair. Regaining the meniscal function by the wrapping treatment prevented cartilage degeneration.

G protein subunit ß1 is an important mediator of the late stage of endochondral ossification.

G protein signaling plays important roles in skeletal development. G protein subunit ß1 (GNB1) is a component of the G protein complex and is associated with G protein signaling. In humans, GNB1 mutations cause global developmental and persistent growth delays and severe neurodevelopmental disability. Similarly, Gnb1-knockout (KO) mice display growth retardation with neural tube defects. These genetic studies raise the possibility that GNB1 regulates skeletal development. This study was designed to investigate the role of GNB1 in skeletal development using Gnb1-KO mice. Gnb1-KO mice showed dwarfism, shortening of limbs, and a decreased ossifying zone of long bones. In situ hybridization and RT-qPCR analyses revealed that Col10a1 and Mmp13 expression was reduced in long bones of Gnb1-KO mice, while Runx2, Osterix, Ihh, and Ppr expression levels were similar to those in wild-type littermates. Gnb1-KO-derived osteoblasts maintained calcification abilities and the expression levels of osteoblast marker genes were unaltered, indicating that osteoblast differentiation and function were not affected in Gnb1-KO mice. Taken together, our results show that GNB1 is required for the late stage of endochondral bone formation by regulating Col10a1 and Mmp13 expression.

Role of interleukin-1 and inflammasomes in oral disease.

BACKGROUND: Inflammation promotes immune cell infiltration into tissues and induces production of pro-inflammatory cytokines that mediate innate immune responses. Acute or temporary inflammation results in the required repair of the inflamed tissues. However, chronic inflammation leads to pathogenesis of inflammatory conditions such as periodontal disease. In periodontal tissues, pro-inflammatory cytokines mediate inflammatory responses and accelerate the bone-resorbing activity of osteoclasts, resulting in destruction of alveolar bone. Levels of interleukin-1 (IL-1), a major pro-inflammatory cytokine that strongly promotes osteoclastic activity, are elevated in oral tissues of patients with periodontitis. Therefore, elucidation of the mechanisms underlying IL-1 production will enhance our understanding of the pathogenesis of periodontal disease. HIGHLIGHT: IL-1 has two isoforms: IL-1α and IL-1ß. Both isoforms bind to the same IL-1 receptor and have identical biological activity. Unlike that of IL-1α, the IL-1ß precursor is not bioactive. To induce its bioactivity, the IL-1ß precursor is cleaved by caspase-1, whose activation is mediated by multiprotein complexes termed inflammasomes. Thus, IL-1ß maturation and activity are strictly regulated by inflammasomes. This review highlights the current understanding of the molecular mechanisms underlying IL-1 production and the related inflammasome activity. CONCLUSION: Inhibition of IL-1 production or the inflammasomes via their regulatory mechanisms may facilitate prevention or treatment of periodontal disease and other inflammatory diseases.

Role of Signal Transduction Pathways and Transcription Factors in Cartilage and Joint Diseases.

: Osteoarthritis and rheumatoid arthritis are common cartilage and joint diseases that globally affect more than 200 million and 20 million people, respectively. Several transcription factors have been implicated in the onset and progression of osteoarthritis, including Runx2, C/EBPß, HIF2α, Sox4, and Sox11. Interleukin-1 ß (IL-1ß) leads to osteoarthritis through NF-ĸB, IκBζ, and the Zn2+-ZIP8-MTF1 axis. IL-1, IL-6, and tumor necrosis factor α (TNFα) play a major pathological role in rheumatoid arthritis through NF-ĸB and JAK/STAT pathways. Indeed, inhibitory reagents for IL-1, IL-6, and TNFα provide clinical benefits for rheumatoid arthritis patients. Several growth factors, such as bone morphogenetic protein (BMP), fibroblast growth factor (FGF), parathyroid hormone-related protein (PTHrP), and Indian hedgehog, play roles in regulating chondrocyte proliferation and differentiation. Disruption and excess of these signaling pathways cause genetic disorders in cartilage and skeletal tissues. Fibrodysplasia ossificans progressive, an autosomal genetic disorder characterized by ectopic ossification, is induced by mutant ACVR1. Mechanistic target of rapamycin kinase (mTOR) inhibitors can prevent ectopic ossification induced by ACVR1 mutations. C-type natriuretic peptide is currently the most promising therapy for achondroplasia and related autosomal genetic diseases that manifest severe dwarfism. In these ways, investigation of cartilage and chondrocyte diseases at molecular and cellular levels has enlightened the development of effective therapies. Thus, identification of signaling pathways and transcription factors implicated in these diseases is important.

Effects of denture adhesives on growth and morphological transformation of Candida albicans.

PURPOSE: We investigated the effects of denture adhesives (cream (Cr), powder (Po), and cushion (Cu)) on growth and adhesive-related morphological transformation of Candida albicans. For this purpose, the numbers of adherent C. albicans, hyphae-specific gene expressions, and the SEM images were examined. METHODS: Acrylic resin blocks were prepared as controls (Co). Cr, Po, and Cu were thinly spread on the surface of the resin block.C. albicans suspension was seeded on the specimens and incubated at 4 °C for 2 h. The numbers of C. albicans adhering to each specimen at each incubation time period (1, 2, 3, 6, 12, and 24 h) were quantified using real-time RT-PCR. The hyphae-specific genes expressions were examined. The surface of each specimen was observed under the SEM to detect the transformation to the hyphal form. RESULTS: The initial adhesion rates in all groups were not statistically significant. The numbers of C. albicans adhering increased with time in all groups, and those adhering to the Cr, Po, and Cu were significantly greater than that adhering to the Co. In the Cr and Po, the hyphal-specific genes expressions were higher after incubation for 6 h. The transformation to the hyphal form was identified in the Cr and Po after incubation for 6 and 12 h. CONCLUSIONS: The denture adhesives used in this study accelerated the growth of C. albicans. Moreover, the early transformation to the hyphal form on the Cr- and Po-type adhesives was observed, suggesting that we should carefully use Cr- and Po-type adhesives.

Evaluation of Meniscal Regeneration in a Mini Pig Model Treated With a Novel Polyglycolic Acid Meniscal Scaffold.

BACKGROUND: Meniscal injury is a severe impediment to movement and results in accelerated deterioration of the knee joint. PURPOSE: To evaluate the effect of a novel meniscal scaffold prepared from polyglycolic acid coated with polylactic acid/caprolactone on the treatment of meniscal injury in a mini pig model. STUDY DESIGN: Controlled laboratory study. METHODS: The model was established with a 10-mm resection at the anterior medial meniscus on both knee joints. A scaffold was implanted in the right knee joint. The meniscal scaffold was inserted and sutured next to the native meniscus. The histological analysis was performed to determine meniscal regeneration with safranin O staining, cell proliferation with PCNA, inflammation with TNF, and collagen structure and production with picrosirius red and immunofluorescence. Cartilage degeneration was evaluated with Safranin O. Meniscal regeneration and joint fluid were evaluated with magnetic resonance imaging. RESULTS: Although compressive stress and elastic modulus were significantly lower in the scaffold than in the native porcine menisci, ultimate tensile stress was similar. Implanted scaffolds were covered with tissue beginning at 4 weeks, with increased migration of proliferating cells to the implant area at 4 and 8 weeks. Scaffolds were absorbed with freshly produced collagen at 24 weeks. Cartilage degeneration was significantly lower in the meniscus-implanted group than in the meniscectomy group. Magnetic resonance imaging results did not show severe accumulation of joint fluids, suggesting negligible inflammation. Density of the implanted menisci was comparable with that of the native menisci. CONCLUSION: Meniscal scaffold prepared from polyglycolic acid has therapeutic potential for meniscal regeneration. CLINICAL RELEVANCE: This meniscal scaffold can improve biological knee reconstruction and prevent the increase of total knee arthroplasty.

Cutting Edge: G Protein Subunit ß 1 Negatively Regulates NLRP3 Inflammasome Activation.

The NLRP3 inflammasome has important roles in the pathogenesis of various inflammatory diseases. However, the regulatory mechanisms of the NLRP3 inflammasome are not fully understood. In this study, we attempted to identify molecules that interact with NLRP3 upon its activation. We identified G protein subunit ß 1 (GNB1), a downstream molecule of G protein-coupled receptors (GPCRs), which regulates the NLRP3 inflammasome activation. GNB1 was physically associated with NLRP3 via the pyrin domain of NLRP3. Activation of the NLRP3 inflammasome was enhanced in GNB1-knockdown or GNB1-deficient murine macrophages, although a lack of GNB1 did not affect activation of the AIM2 inflammasome. ASC oligomerization induced by NLRP3 was enhanced by GNB1 deficiency. Conversely, NLRP3-dependent ASC oligomerization was inhibited by the overexpression of GNB1. This study indicates that GNB1 negatively regulates NLRP3 inflammasome activation by suppressing NLRP3-dependent ASC oligomerization, and it provides a regulatory mechanism of the NLRP3 inflammasome.

Hyaluronic acid promotes proliferation and migration of human meniscus cells via a CD44-dependent mechanism.

PURPOSE: Treatment of meniscal injury is important for osteoarthritis (OA) prevention. Meniscus cells are divided between inner and outer cells, which have different characteristics and vascularity. We evaluated the effects of hyaluronic acid (HA) on the proliferation and migration of human inner and outer meniscus cells, and investigated the underlying healing mechanisms. MATERIALS AND METHODS: Lateral menisci from 18 patients who underwent total knee arthroplasty were used. Meniscus cells were harvested from the outer and inner menisci and evaluated using migration and proliferation assays after treatment with HA or chondroitin sulfate (CS). The effects of HA on prostaglandin E2 (PGE2)-induced apoptosis and gene expression were evaluated. RESULTS: Cell migration and proliferation were increased by HA in a concentration-dependent manner, in both inner and outer meniscus cells. PGE2-induced apoptosis and caspase-3/7 activity were suppressed by HA in both inner and outer meniscus cells, and these effects were blocked by an anti-CD44 antibody. COL2A1 and ACAN mRNA levels were upregulated following HA treatment of inner meniscus cells. MMP13 mRNA was downregulated following CS stimulation of both inner and outer meniscus cells. These results suggest that CS treatment suppresses the inflammatory reaction rather than providing meniscal restoration. The phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways were activated by HA in both types of meniscus cells; these effects were blocked by treatment with an anti-CD44 antibody. CONCLUSIONS: HA promoted human meniscus regeneration by inhibiting apoptosis, promoting cell migration, and accelerating cell proliferation, potentially through the PI3K/MAPK pathway via the CD44 receptor.

Sox4 is involved in osteoarthritic cartilage deterioration through induction of ADAMTS4 and ADAMTS5.

Osteoarthritis is a common disease in joint cartilages. Because the molecular pathogenesis of osteoarthritis remains elusive, early diagnostic markers and effective therapeutic agents have not been developed. To understand the molecular mechanisms, we attempted to identify transcription factors involved in the onset of osteoarthritis. Microarray analysis of mouse articular cartilage cells indicated that retinoic acid, a destructive stimulus in articular cartilage, up-regulated expression of sex-determining region Y-box (Sox)4, a SoxC family transcription factor, together with increases in Adamts4 and Adamts5, both of which are aggrecanases of articular cartilages. Overexpression of Sox4 induced a disintegrin-like and metallopeptidase with thrombospondin type 4 and 5 motif (ADAMTS4 and ADAMTS5, respectively) expression in chondrogenic cell lines C3H10T1/2 and SW1353. In addition, luciferase reporter and chromatin immunoprecipitation assays showed that Sox4 up-regulated ADAMTS4 and Adamts5 gene promoter activities by binding to their gene promoters. Another SoxC family member, Sox11, evoked similar effects. To evaluate the roles of Sox4 and Sox11 in articular cartilage destruction, we performed organ culture experiments using mouse femoral head cartilages. Sox4 and Sox11 adenovirus infections caused destruction of articular cartilage associated with increased Adamts5 expression. Finally, SOX4 and SOX11 mRNA expression was increased in cartilage of patients with osteoarthritis compared with nonosteoarthritic subjects. Thus, Sox4, and presumably Sox11, are involved in osteoarthritis onset by up-regulating ADAMTS4 and ADAMTS5.-Takahata, Y., Nakamura, E., Hata, K., Wakabayashi, M., Murakami, T., Wakamori, K., Yoshikawa, H., Matsuda, A., Fukui, N., Nishimura, R. Sox4 is involved in osteoarthritic cartilage deterioration through induction of ADAMTS4 and ADAMTS5.

Hybrid high tibial osteotomy is superior to medial opening high tibial osteotomy for the treatment of varus knee with patellofemoral osteoarthritis.

PURPOSE: To evaluate patellofemoral congruity after opening wedge high tibial osteotomy (OWHTO) and hybrid HTO. METHODS: Twenty-four knees with hybrid HTO and 24 with OWHTO were evaluated in this study. The Caton-Deschamps and modified Miura-Kawamura indices were used to evaluate pre- and post-operative patellar heights for both types of surgery. Tibial tuberosity-trochlear groove (TT-TG) distance, patellar tilt, and medial and lateral joint space at the patellofemoral joint were compared. Anterior knee pain was assessed using the Kujala anterior knee pain scale. RESULTS: There was no significant difference between the correction angles of the hybrid HTO and OWHTO. Pre- and post-operative values for the Caton-Deschamps and modified Miura-Kawamura indices in patients who underwent hybrid HTO changed from 0.90 to 0.94 and from 0.95 to 1.03, respectively, with no significant differences noted. Following OWHTO, these values decreased significantly from 0.91 to 0.73 and from 1.06 to 0.84, respectively (p < 0.01). The post-operative patellar height after OWHTO was significantly lower than that after hybrid HTO (p < 0.01). After hybrid HTO, the TT-TG distance decreased significantly from 11.4 to 7.4 (p < 0.01), but it did not change significantly after OWHTO. Although pre- and post-operative patellar tilt were not altered significantly in either group, the medial joint space of the patellofemoral joint was significantly increased post-operatively following hybrid HTO (p = 0.035). The pre-operative Kujala scores were significantly lower in the hybrid HTO group, but post-operative scores improved in both groups. CONCLUSIONS: Hybrid HTO provides a better post-operative patellofemoral joint than does OWHTO with regard to patellar position and reduction of the TT-TG distance, as well as improved clinical outcomes. Hybrid HTO, rather than OWHTO, is the preferred technique for the treatment of varus knees combined with patellofemoral osteoarthritis. LEVEL OF EVIDENCE: Retrospective comparative study, Level III.