RESUMO
Transplanted mesenchymal stromal cells (MSCs) exhibit a robust anti-inflammatory and homing capacity in response to high inflammatory signals, as observed in studies focused on rheumatic diseases that target articular cartilage (AC) health. However, AC degradation in osteoarthritis (OA) does not necessarily coincide with a highly inflammatory joint profile. Often, by the time patients seek medical attention, they already have damaged AC. In this study, we examined the therapeutic potential of a single bone marrow MSC transplant (2 × 106 cells/kgbw) through two different routes: intra-articular (MSCs-IAt) and intravenous (MSCs-IVt) in a preclinical model of low-grade inflammatory OA with an established AC degeneration. OA was induced through the destabilization of the medial meniscus (DMM) in female Wistar Kyoto rats. The animals received MSCs 9 weeks after surgery and were euthanized 4 and 12 weeks post-transplant. In vivo and ex vivo tracking of MSCs were analyzed via bioluminescence and imaging flow cytometry, respectively. Cytokine/chemokine modulation in serum and synovial fluid was measured using a multiplex panel. AC degeneration was quantified through histology, and hindlimb muscle balance was assessed with precision weighing. To our knowledge, we are the first group to show the in vivo (8 h) and ex vivo (12 h) homing of cells to the DMM-OA joint following MSCs-IVt. In the case of MSCs-IAt, the detection of cellular bioluminescence at the knee joint persisted for up to 1 week. Intriguingly, intra-articular saline injection (placebo-IAt) resulted in a worse prognosis of OA when compared to a non-invasive control (placebo-IVt) without joint injection. The systemic cytokines/chemokines profile exhibited a time-dependent variation between transplant routes, displaying a transient anti-inflammatory systemic response for both MSCs-IVt and MSCs-IAt. A single injection of MSCs, whether administered via the intra-articular or intravenous route, performed 9 weeks after DMM surgery, did not effectively inhibit AC degeneration when compared to a non-invasive control.
Assuntos
Cartilagem Articular , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Osteoartrite , Humanos , Ratos , Feminino , Animais , Meniscos Tibiais/metabolismo , Osteoartrite/metabolismo , Cartilagem Articular/metabolismo , Anti-Inflamatórios/farmacologia , Injeções Intra-Articulares , Células-Tronco Mesenquimais/metabolismo , Transplante de Células-Tronco Mesenquimais/métodosRESUMO
Osteoarthritis is the most common form of arthritis among elderly adults. Herein, we performed protein-protein interaction (PPI) and miRNA network analysis to evaluate the global correlation between miRNA regulation and the PPI network in human osteoarthritis. Our results showed that desmoplakin (DSP), cystatin A (CSTA), calmodulin 1, tyrosine kinase endothelial, insulin-like growth factor 1 (IGF-1), IGF-binding protein 7 (IGFBP7), syndecan 1 (SDC1), ephrin type-A receptor 4, and PDZ and LIM domain protein 1 were associated with osteoarthritis. Among these proteins, DSP and CSTA interaction and IGF-1, IGFBP7 and SDC1 interaction were observed in our PPI network. Furthermore, these potential target proteins were also linked with individual miRNA in the network. Our findings shed light on the PPIs and mechanisms by which miRNA may regulate the protein interaction network in osteoarthritis, which might provide theoretical support for further studies aimed at discovering new therapeutic strategies.
Assuntos
MicroRNAs/genética , Osteoartrite/genética , Osteoartrite/metabolismo , Mapas de Interação de Proteínas , Cistatina A/metabolismo , Desmoplaquinas/metabolismo , Redes Reguladoras de Genes , Humanos , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Meniscos Tibiais/metabolismo , Meniscos Tibiais/patologia , Modelos Biológicos , Modelos Genéticos , Ligação Proteica , Mapeamento de Interação de Proteínas , Transdução de Sinais/genética , Sindecana-1/metabolismoRESUMO
We described the behaviour of 120 days rabbit knee-meniscus cells in monolayer culture. The cells were grown forming cellular aggregates resembling true cellular nodules. Three stages of development of these nodules could be observed: formation of the cellular nodules between days 1 and 3; nodular growth, with their maximal at day 5; and nodular regression beginning at day 8. Ultrastructural analysis of the extracellular matrix of these cellular nodules was assessed on days 3, 5 and 8. At the formation stage, we could observe striated collagen fibrils and small bundles of tubular microfibrils either interspersed with very low quantities of amorphous elastin, being morphologically identical to elaunin fibers, or without only trace of elastin, being morphologically identical to oxytalan fibers. By day 5, fibrillar elements with 100 nm periodic ladder-like collagen VI fibrillar aggregates could also be detected. At day 8, the striated collagen fibrils and oxytalan fibers could not be observed. During this same period, there was an increase of a dense matrix comprised of collagen VI and mature elastic fibers. Chondroitin/dermatan sulfate proteoglycans were synthesized and became essential for the arrangement of collagen type VI, since chondroitinase ABC treatment of the culture disrupted collagen VI assembly, associated with the large spaces near the cell surface. In addition, the cells lost their fusiform morphology and changed into rounded cells. The results show that primary cultures of rabbit meniscus fibrochondrocytes maintain their capacity to form chondro-like structures in vitro. The organization process was rapid and uniform throughout the entire culture presuming that the normal signal transduction pathways are maintained intact and that essential factors in some phases of tissue organization are present.