Cartilage injuries often represent irreversible tissue damage because cartilage has only a low ability to regenerate. Thus, cartilage loss results in permanent damage, which can become the starting point for osteoarthritis. In the past, bioactive glass scaffolds have been developed for bone replacement and some of these variants have also been colonized with chondrocytes. However, the hydroxylapaptite phase that is usually formed in bioglass scaffolds is not very suitable for cartilage formation (chondrogenesis). This interdisciplinary project was undertaken to develop a novel slowly degrading bioactive glass scaffold tailored for cartilage repair by resembling the native extracellular cartilage matrix (ECM) in structure and surface properties. When colonized with articular chondrocytes, the composition and topology of the scaffolds should support cell adherence, proliferation and ECM synthesis as a prerequisite for chondrogenesis in the scaffold. To study cell growth in the scaffold, the scaffolds were colonized with human mesenchymal stromal cells (hMSCs) and primary porcine articular chondrocytes (pACs) (27,777.8 cells per mm3) for 7 – 35 d in a rotatory device. Cell survival in the scaffold was determined by vitality assay. Scanning electron microscopy (SEM) visualized cell ultramorphology and direct interaction of hMSCs and pACs with the bioglass surface. Cell proliferation was detected by CyQuant assay. Subsequently, the production of sulphated glycosaminoglycans (sGAGs) typical for chondrogenic differentiation was depicted by Alcian blue staining and quantified by dimethylmethylene blue assay assay. Quantitative real-time polymerase chain reaction (QPCR) revealed gene expression of cartilage-specific aggrecan, Sox9, collagen type II and dedifferentiation-associated collagen type I. To demonstrate the ECM-protein synthesis of the cells, the production of collagen type II and type I was determined by immunolabelling. The bioactive glass scaffold remained stable over the whole observation time and allowed the survival of hMSCs and pACs for 35 days in culture. The SEM analyses revealed an intimate cell-biomaterial interaction for both cell types showing cell spreading, formation of numerous filopodia and ECM deposition. Both cell types revealed initial proliferation, decreasing after 14 days and becoming elevated again after 21 days. hMSCs formed cell clusters, whereas pACs showed an even distribution. Both cell types filled more and more the pores of the scaffold. The relative gene expression of cartilage-specific markers could be proven for hMSCs and pACs. Cell associated sGAGs deposition could be demonstrated by Alcian blue staining and sGAGs were elevated in the beginning and end of the culturing period. While the production of collagen type II could be observed with both cell types, the synthesis of aggrecan could not be detected in scaffolds seeded with hMSCs. hMSCs and pACs adhered, spread and survived on the novel bioactive glass scaffolds and exhibited a chondrocytic phenotype.
The superficial zone (SFZ) of articular cartilage has unique structural and biomechanical features, and is important for joint long-term function. Previous studies have shown that TGF-β/Alk5 signaling upregulating PRG4 expression maintains articular cartilage homeostasis. However, the exact role and molecular mechanism of TGF-β signaling in SFZ of articular cartilage homeostasis are still lacking. In this study, a combination of in vitro and in vivo approaches were used to elucidate the role of Alk5 signaling in maintaining the SFZ of articular cartilage and preventing osteoarthritis initiation. Mice with inducible cartilage SFZ-specific deletion of Alk5 were generated to assess the role of Alk5 in OA development. Alterations in cartilage structure were evaluated histologically. The chondrocyte apoptosis and cell cycle were detected by TUNEL and Edu staining, respectively. Isolation, culture and treatment of SFZ cells, the expressions of genes associated with articular cartilage homeostasis and TGF-β signaling were analyzed by qRT-PCR. The effects of TGF-β/Alk5 signaling on proliferation and differentiation of SFZ cells were explored by cells count and alcian blue staining. In addition, SFZ cells isolated from C57 mice were cultured in presence of TGF-β1 or SB505124 for 7 days and transplanted subcutaneously in athymic mice. Postnatal cartilage SFZ-specific deletion of Alk5 induced an OA-like phenotype with degradation of articular cartilage, synovial hyperplasia as well as enhanced chondrocyte apoptosis, overproduction of catabolic factors, and decreased expressions of anabolic factors in chondrocytes. qRT-PCR and IHC results confirmed that Alk5 gene was effectively deleted in
Introduction: Osteoarthritis is the most common joint disease in the world. Biochemical and genetic factors as well as mechanical stress contribute to lesions in the cartilage. The present study analyses the effect of b-Endorphin on the
Purpose: Intraarticular use of anaesthetic agents is common for postoperative pain relief after arthroscopic knee surgery. In this study, we have evaluated and compared the effects of Bupivacaine, Levobupivacaine and Tramadol both invivo and invitro experimental rat models on articular cartilage and chondrocytes. Materials and Methods: Invivo Experiment: 1. Injections: Thirty mature Sprague-Dawley rats weighing 230 – 300 g were randomized into 3 groups. Bupivacaine (Group 1), Levobupivacaine (Group 2) and Tramadol (Group 3) were injected into the right knee joints and physiological 0.9% saline into the left. 2. Histopathologic Analysis: The specimens were fixed, decalcified and stained with Hematoxylen and Eosin (H&
E) and Toluidin Blue. All slides were examined by the same pathologist, who was blinded to the injectate used in each joint. All samples were evaluated histopathologically according to the recommendation of International Cartilage Repair Society’s osteoarthritis and cartilage histopathology grading and staging system. Invitro Experiment:
Background: To date, conventional freezing and cryopreservation of articular cartilage has had limited success due to the mechanical injury of cells resulting from uncontrolled ice crystal propagation. Frozen then thawed grafts show a total lack of viable
This study investigated confocal laser scanning microscopy (CLSM) as a novel method of imaging of chondrocytes on a collagen membrane used for
Unique progenitor cells have been identified recently and successfully cultured in vitro from human
In 1823 J. White excised the head. In 1887 a German surgeon replaced the head with ivory. Interposition arthroplasties were common after WW1. Short-stemmed head replacing prosthesis were developed after WW2. Moores and Thompson designed a more stable intramedullary stem. Acetabular erosion was troublesome—and so replacing both surfaces started in the late 1950s using Teflon cup and metal femur. Unfortunately, these quickly became loose due to wear or sepsis. In 1960, Charnley used a polyethylene cup and stainless-steel femur and fixed both with dental cement. This ‘low friction arthroplast’ became a routine procedure after 1961. In the 1970s there were many ‘Charnley look-alike’ prosthesis with similar problems of poly-wear, granulomas and cysts causing bone loss, loosening, breakages and infection. Resurfacing with two thin shells was developed to reduce the foreign material, the bone resection and the cement used. Unfortunately, neck fractures, avascular necrosis and excessive wear of the poly shell were common. Despite operating theatres with laminar flow of sterile air, space suits and improved cementing techniques, the same problems occurred. To avoid poly and cement, Mittelmayer developed a ceramic screw cup, which did not require cement. Although some screws migrated, they did not wear. Because the un-cemented metal stem remained fixed solid to the femur, un-cemented metal cups and stems were developed. To avoid the poly-wear, ceramic liners became popular. To provide the active patients with a stable joint that requires no restriction in physical activity, a large head in a large cup is desirable. Unfortunately, the large metal-on-metal resurfacing prosthesis produce metal wear ions and nanoparticles which can form hypersensitivities, cysts and pseudotumours. Computer assisted navigation to ensure correct positioning of the prosthetic components is obviously useful for surgeons that use incisions too small to see enough to be certain of the cups position. Presently, articular cartilage research is progressing rapidly and by 2020 most arthritic hip joints will be arthroscopically debrided and resurfaced by an injection of genetically engineered