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Aim: The aim of this study is to prepare and characterize simvastatin-loaded nanoemulsions (SIM-LN) as well as evaluate their physicochemical properties and toxicity. Methodology & results: The SIM-LN were prepared, their characteristics evaluated for 30 days, and after that, the SIM-LN toxicity was evaluated using Vero cell culture and the in vivo model of Caenorhabditis elegans. The prepared SIM-LN had an average droplet size of 139 ± 22 nm, with high encapsulation rate (>98.4%). The storage at room temperature proved to be the most optimal condition. Toxicity assays demonstrated no toxicity. Conclusion: It was demonstrated that the surfactants used as emulsifiers optimized the properties without side effects, because no toxicity was measured in preliminary tests.
RESUMO
Chondrocyte health is altered when exposed to local anesthetics, raising concerns as to the long-term effects of local anesthetics intra-articularly for diagnosis and analgesia. To investigate the drug with the lowest toxic potential, the effect of ropivacaine and mepivacaine on chondrocytes was evaluated. Articular cartilage from normal metacarpophalangeal joints of five equine cadaver specimens was used to establish chondrocyte cultures. Following seven days, chondrocytes were exposed to standard culture medium (DMEM), ropivacaine 7.5 mg/ml (ROP7.5), ropivacaine 10 mg/ml (ROP10), mepivacaine 20 mg/ml (MEP20), mepivacaine 30 mg/ml (MEP 30), and 0.9% saline solution (SAL). Chondrocyte viability was evaluated by trypan blue exclusion, MTT, and flow cytometry via cellular staining with propidium iodide. No differences were observed between treatments following trypan blue exclusion assay. A difference was observed between DMEM and all other treatment groups (P < .0001) with a significant viability drop using the MTT assay. Mepivacaine 20 mg/ml and MEP30 exposure between showed greatest decrease in cellular viability compared to SAL, ROP7.5, and ROP10 (P < .0001). Cellular viability decreased as measured by flow cytometry in all groups compared to DMEM and ROP7.5 (P < .02). Interestingly, the trypan blue, MTT, and flow cytometry assays yielded different results. Although there was no difference using trypan blue, MTT demonstrated that ropivacaine-treated cells had lower viability than DMEM, and cytometry found that ROP7.5 did not differ from DMEM. Results in vitro suggest that short-term exposure to ropivacaine may result in less chondrotoxicity than mepivacaine. In vivo studies are warranted investigating long-term effects of local anesthetics on equine articular cartilage.
Assuntos
Mepivacaína , Ropivacaina , Animais , Bupivacaína , Células Cultivadas , Condrócitos , CavalosRESUMO
ABSTRACT: In healthy cartilage, chondrocytes maintain an expression of collagens and proteoglycans and are sensitive to growth factors and cytokines that either enhance or reduce type II collagen synthesis. In osteoarthritis, pro-inflammatory cytokines, such as IL-6, induce overexpression of metalloproteinases (MMP) and decreasing synthesis of aggrecan. Use of chondroprotectors agents, such as Platelet-Rich Plasma (PRP) and triamcinolone (TA) are alternatives to reduce the progression of joint damage. In this study, we used chondrocytes extracted from metacarpophalangeal joints of healthy horses as the experimental model. Cells were treated in vitro with PRP or TA. No differences were observed between these treatments in comparison to the control group when the expressions of MMP9, MMP13, IL-6 and ACAN genes were evaluated (P<0.05). With these results, we can suggest that the treatments were not deleterious to equine cultured chondrocyte, once they did not stimulate MMPs and IL-6 synthesis or caused changes in ACAN.
RESUMO: Na cartilagem saudável, os condrócitos mantêm a expressão de colágenos e proteoglicanos, sendo sensíveis a fatores de crescimento e citocinas que aumentam ou reduzem a síntese de colágeno tipo II. Na osteoartrite, citocinas pró-inflamatórias, como a IL-6, estimulam a expressão de metaloproteinases (MMP) e reduzem a síntese de agrecano. O uso de condroprotetores, como o Plasma Rico em Plaquetas (PRP) e triancinolona (TA) é uma alternativa para se reduzir a progressão do dano articular. Neste estudo foram usados condrócitos extraídos das articulações metacarpofalangeanas de equinos saudáveis. As células foram tratadas in vitro com TA ou PRP. Não foram observadas diferenças entre os tratamentos comparando-se com o grupo controle quanto à expressão genética de MMP-9, MMP-13, IL-6 e ACAN (p<0,05). Assim, pode-se sugerir que os tratamentos não foram deletérios ao cultivo de condrócitos, uma vez que não estimularam a síntese de MMP e IL-6 e nem causaram alterações no ACAN.
RESUMO
ABSTRACT: Progressive deterioration and loss of articular cartilage are the final degenerative events common to osteoarthritis (OA). Reactive oxygen species (ROS) play an important role in this chondrocyte catabolic activity, leading to cell death and matrix components breakdown. Intra-articular corticosteroid injections such as triamcinolone acetonide have been used to control pain and inflammation associated with OA. New treatments for OA, platelet-rich plasma and pentosan polysulphate sodium have also been used and further investigations are necessary to determine their safety in joint cells. In this in vitro study, the use of these three substances (triamcinolone acetonide, platelet-rich plasma, and pentosan polysulphate sodium) in healthy chondrocytes did not alter the antioxidant status when compared to control groups, indicating that they could be considered safe in healthy conditions.
RESUMO: A deterioração progressiva e perda da cartilagem articular são os eventos finais da osteoartrite (OA). Espécies reativas de oxigênio (ROS) têm papel importante na atividade catabólica de condrócitos, levando a morte celular e quebra dos componentes da matriz. Injeções intra-articulares de corticosteroides, como com o acetonido de triancinolona, são usadas para controle da dor e inflamação associadas à OA. Novos tratamentos para a OA, como o plasma rico em plaquetas e o pentosano polissulfato sódico, também tem sido utilizados e necessitam de maiores investigações para determinar sua segurança para as células articulares de equinos. Neste estudo in vitro, o uso destas três substâncias (acetonido de triancinolona, pentosan polissulfato de sódio de plasma rico em plaquetas) em condrócitos saudáveis de equinos não alterou o status antioxidante quando comparado aos grupos controle, indicando que puderam ser considerados seguros em condições saudáveis.
