BACKGROUND: Exosomes from mesenchymal stem cells (MSCs) show anti-inflammatory effect on osteoarthritis (OA); however, their biological effect and mechanism are not yet clearly understood. This study investigated the anti-inflammatory effect and mechanism of MSC-derived exosomes (MSC-Exo) primed with IL-1b in osteoarthritic SW982 cells. METHODS: SW982 cells were treated with interleukin (IL)-1b and tumor necrosis factor (TNF)-a to induce the OA phenotype. The effect of exosomes without priming (MSC-Exo) or with IL-1b priming (MSC-IL-Exo) was examined on the expression of pro-or anti-inflammatory factors, and the amount of IjBa was examined in SW982 cells. Exosomes were treated with RNase to remove RNA. The role of miR-147b was examined using a mimic and an inhibitor. RESULTS: MSC-IL-Exo showed stronger inhibitory effects on the expression of pro-inflammatory cytokines (IL-1b, IL-6, and monocyte chemoattractant protein-1) than MSC-Exo. The expression of anti-inflammatory factors (SOCS3 and SOCS6) was enhanced by MSCs-IL-Exo. Priming with IL-1b increased RNA content in MSC-IL-Exo, and pretreatment with RNase abolished anti-inflammatory effect in SW982 cells. miR-147b was found in much larger amounts in MSC-IL-Exo than in MSC-Exo. The miR-147b mimic significantly inhibited the expression of inflammatory cytokines, while the miR-147b inhibitor only partially blocked the anti-inflammatory effect of MSC-IL-Exo. MSC-IL-Exo and miR-147b mimic inhibited the reduction of IjBa, an nuclear factor kappa-light-chain-enhancer of activated B cells (NF-jB) inhibitor, by IL-1b and TNF-a. CONCLUSION: This study showed that MSC exosomes with IL-1b priming exhibit significantly enhanced anti-inflammatory activity in osteoarthritic SW982 cells. The effect of IL-1b-primed MSC exosomes is mediated by miRNAs such as miR-147b and involves inhibition of the NF-jB pathway.
Cartilage extracellular matrix contains antiadhesive and antiangiogenic molecules such as chondromodulin‐1, thrombospondin‐1, and endostatin. We have aimed to develop a cross‐linked cartilage acellular matrix (CAM) barrier for peritendinous adhesion prevention. CAM film was fabricated using decellularized porcine cartilage tissue powder and chemical cross‐linking. Biochemical analysis of the film showed retention of collagen and glycosaminoglycans after the fabrication process. Physical characterization of the film showed denser collagen microstructure, increased water contact angle, and higher tensile strength after cross‐linking. The degradation time in vivo was 14 d after cross‐linking. The film extract and film surface showed similar cell proliferation, while inhibiting cell migration and cell adhesion compared to standard media and culture plate, respectively. Application of the film after repair resulted in similar tendon healing and significantly less peritendinous adhesions in a rabbit Achilles tendon injury model compared to repair only group, demonstrated by histology, ultrasonography, and biomechanical testing. In conclusion, the current study developed a CAM film having biological properties of antiadhesion, together with biomechanical properties and degradation profile suitable for prevention of peritendinous adhesions.
Purpose: To compare cell yield and character of synovium-derived mesenchymal stem cell (SDMSC) harvested by 2 different techniques using rongeur and motorized shaver during knee arthroscopy. Methods: This study was performed in 15 patients undergoing partial meniscectomy. Two different techniques were used to harvest SDMSCs in each patient from the synovial membrane at 2 different locations overlying the anterior fat pad, each within 1 minute of harvest time. Cell yield and proliferation rates were evaluated. Cell surface marker analysis was done after passage 2 (P2). Trilineage differentiation potential was evaluated by real-time quantitative polymerase chain reaction and histology. Statistical analysis between the 2 methods was done using the ManneWhitney U test. Results: Wet weight of total harvested tissue was 69.93 (AE 20.02) mg versus 378.91 (AE 168.87) mg for the rongeur and shaver group, respectively (P < .0001). Mononucleated cell yield was 3.32 (AE 0.89) versus 3.18 (AE 0.97) Â 10 3 cells/mg, respectively (P ¼ .67). Fluorescence-activated cell sorting analysis revealed similar SDMSC-related cell surface marker expression levels in both groups, with positive expression for CD44, CD73, CD90, and CD105 and decreased expression for CD34 and CD45. Both groups showed similar trilineage differentiation potential in histology. Chondrogenic (SOX9, ACAN, COL2), adipogenic (LPL, PLIN1, PPAR-g), and osteogenic (OCN, OSX, RUNX2) gene marker expression levels also were similar between both groups. Conclusions: No difference was observed between rongeur biopsy and motorized shaver harvest methods regarding SDMSC yield and cell characteristics. Clinical Relevance: The current study shows that both rongeur and motorized shaver harvest are safe and effective methods for obtaining SDMSCs. Motorized shaver harvest results in higher volume of tissue acquisition per time, thereby leading to higher number of SDMSCs which may be useful during clinical application.
Thermo-responsive sol-gel was prepared from methylcellulose (MC) and xanthan gum (XT) for adhesion barrier. The prepared MC/XT sol-gel showed transition temperature and gelation around body temperature (37 o C) within 1 min. In vivo biocompatibility and biodegradability of the MC/XT sol-gel were validated by implantation of the gel in rats. Also, the MC/XT sol-gel was highly effective for the prevention of tissue and organ adhesion. Therefore, the MC/ XT sol-gel can be a good candidate material as a tissue adhesion barrier.
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