Insulin-Transferrin-Selenium Prevent Human Chondrocyte Dedifferentiation And Promote The Formation Of High Quality Tissue Engineered Human Hyaline Cartilage

This study was to investigate the effects of Insulin-Transferrin-Selenium (ITS) on the proliferation and quantitative gene expression of adult human nasal septum chondrocytes in monolayer culture expansion and the formation of tissue engineered hyaline cartilage. Effects of ITS on human nasal septum chondrocytes monolayer culture expansion and gene expression were evaluated in various culture media either added with 2% fetal bovine serum (FBS) or 1ng/mL basic fibroblast growth factor plus 1ng/mL transforming growth factor or both serum and growth factors supplementation in comparison with medium added with 10%FBS. Chondrocytes cultured in medium added with 2% fetal bovine serum and growth factors either supplemented with or without ITS were then mixed with pluronic F-127 hydrogel for in vivo tissue engineered cartilage formation in nude mice model. Engineered tissues were removed after 8 weeks of implantation and evaluated with histological staining, immunohistochemistry, transmission electron microscopy and quantitative gene expression analysis. ITS promoted human chondrocytes proliferation and reduced chondrocytes dedifferentiation in media supplemented with serum and growth factors. ITS with 2% FBS and growth factors provided 15-fold increased in chondrocytes number by the end of the culture period compared to the standard culture medium used in chondrocytes culture (medium added with 10% FBS). Engineered tissue resulted from ITS supplementation demonstrated higher quality of cartilage formation. In conclusion, our study has demonstrated the benefits of ITS supplementation in human chondrocytes monolayer culture and tissue engineering cartilage formation

Intraarticular injection of autologous chondrogenic induced bone marrow stem cells promotes regenesis of traumatic osteoarthritis and promotes repair of meniscus injury in a sheep model

Objective. The aim of this study was to determine if autologous bone marrow mesenchymal stem cells (AMSCs) cultured in chondrogenic medium containing growth factors could repair meniscus tissue and retard surgically induced osteoarthritis in the knee joint. Methods. Autologous Sheep bone marrow stem cells (AMSCs) were aspirated from sheep iliac crest and cultured in medium containing 5ng/ml TGFβ3 + 50ng/ml IGF-1 for 3 weeks. Six weeks after surgically induced osteoarthritis; a single dose of chondrogenic inducted AMSCs suspended in 5 ml Dulbecco’s Modified Eagle Medium (DMEM) at density of 10 million cells was injected intra-articularly into the osteoarthritic knee joint. After addition of another 6 weeks, the sheep were scarified and the femoral condyle and the proximal tibia were removed, fixed and photographed. The gross of the removed knee joint were graded by a blinded assessor based on ICRS grading system. Part of the tissue was imbedded in paraffin and sectioned into 5µm and stained with H&E and Safranin O. Result. The results indicated that surgically induced osteoarthritic knee joint treated with chondrogenic inducted AMSCs showed clear evidence of minimum articular cartilage destruction with meniscus repaired when compared with other control knee joints. Conclusion Our findings demonstrated that intra-articular injection of chondrogenic inducted AMSCs could stimulate regeneration of meniscal tissue and retards the progressive of osteoarthritis in sheep model

Anterior cruciate ligament resection and medial meniscectomy result in multifocal cartilage degenerations

Abstract: Damage to the meniscus and the anterior cruciate ligament alters the pattern of loading in the knee joint and frequently leads to cartilage degeneration and osteoarthritis. This study aimed to evaluate the area of greatest impact after traumatic osteoarthritic induction at the knee joint in sheep model. Osteoarthritis was induced unilaterally at the right hind knee of six sheep by surgical resection of the anterior cruciate ligament and medial meniscus. After 3 weeks recovery period, sheep were exercised once daily by running a 100 meters distance on a hard surface for 3 weeks. Gross and histological assessments were done. Six regions examined were; the patella, patella femoral groove, medial femoral condyle, lateral femoral condyle, medial tibia plateau and lateral tibia plateau. Their mean International Cartilage Repair Society grading was: 2.5±0.42, 3.3±0.17, 2.25±0.38, 1.33±0.25, 2±0.37 and 1.5±0.22 respectively. The control from all samples scored zero at the above regions. The lesions at various regions were significantly higher compared to the control (p < 0.05). Patella femoral groove was significantly higher compare to all other regions except for Patella. Lateral femoral condyle was the least (p < 0.05). Histological staining revealed defined osteoarthritic changes. Total excision of medial meniscus and anterior cruciate ligament followed by exercise regime induced osteoarthritis in the affected knee joint. The changes in pattern of degeneration, affected patella femoral groove the most; followed by the patella and medial femoral condyle, then medial tibia plateau, lateral tibia plateau and lastly the lateral femoral condyle