Studies have been conducted to highlight the anti-inflammatory and immunosuppressive properties of cannabinoids and also their potentials for cartilage repair and regeneration. Various wound healing techniques can be used to investigate the mechanisms of chondrocyte repair in monolayers or three dimensional tissue constructs. The effect of different concentrations of the synthetic cannabinoids WIN55, 212-2 (WIN-2), URB602 and HU-308 with and without their antagonists on the wound healing of chondrocyte monolayers was investigated using a simple scratch assay model. The three cannabinoids were found to increase wound healing of chondrocyte monolayers, but at different rates. WIN55, 212-2 at a concentration of 1μM had the highest effect of increasing both migration and proliferation of chondrocytes cultured in a chondrogenic media, which increased the rate of wound closure. It was also found that treating the cells with 2μM of any of the cannabinoids lead to a decrease in cell proliferation and the rate of wound closure. These findings were further investigated, by studying the effect of WIN-2 on nitric oxide (NO) and matrix metalloproteinase-2 (MMP-2) expressed by wounded chondrocyte monolayers. Moreover, expression of collagen type-I, collagen type-II, fibronectin and S100 proteins were detected using immunofluorescence and verified quantitatively using ELISA based techniques, following treatment with 1μM and 2μM of WIN-2, for both 2D monolayers and 3D sheets. Treating chondrocytes with 1μM of WIN-2 significantly increased collagen type-II, fibronectin and S100, and significantly reduced collagen type-I compared to control groups in monolayers and chondrocyte cell sheets. On the other hand, both concentrations of WIN-2 significantly reduced the expression of the inflammation markers NO, and MMP-2, in a dose dependent manner. These findings highlight the potential use of the synthetic cannabinoid for improving the rate of wound closure as well as acting as an antiinflammatory agent, which could be used to enhance tissue engineering protocols aimed at cartilage repair.

Studies have been conducted to highlight the anti-inflammatory and immunosuppressive properties of cannabinoids and also their potentials for cartilage repair and regeneration. Various wound healing techniques can be used to investigate the mechanisms of chondrocyte repair in monolayers or three dimensional tissue constructs. The effect of different concentrations of the synthetic cannabinoids WIN55, 212-2 (WIN-2), URB602 and HU-308 with and without their antagonists on the wound healing of chondrocyte monolayers was investigated using a simple scratch assay model. The three cannabinoids were found to increase wound healing of chondrocyte monolayers, but at different rates. WIN55, 212-2 at a concentration of 1μM had the highest effect of increasing both migration and proliferation of chondrocytes cultured in a chondrogenic media, which increased the rate of wound closure. It was also found that treating the cells with 2μM of any of the cannabinoids lead to a decrease in cell proliferation and the rate of wound closure. These findings were further investigated, by studying the effect of WIN-2 on nitric oxide (NO) and matrix metalloproteinase-2 (MMP-2) expressed by wounded chondrocyte monolayers. Moreover, expression of collagen type-I, collagen type-II, fibronectin and S100 proteins were detected using immunofluorescence and verified quantitatively using ELISA based techniques, following treatment with 1μM and 2μM of WIN-2, for both 2D monolayers and 3D sheets. Treating chondrocytes with 1μM of WIN-2 significantly increased collagen type-II, fibronectin and S100, and significantly reduced collagen type-I compared to control groups in monolayers and chondrocyte cell sheets. On the other hand, both concentrations of WIN-2 significantly reduced the expression of the inflammation markers NO, and MMP-2, in a dose dependent manner. These findings highlight the potential use of the synthetic cannabinoid for improving the rate of wound closure as well as acting as an antiinflammatory agent, which could be used to enhance tissue engineering protocols aimed at cartilage repair. / Egyptian Government

Made available in DSpace on 2017-07-24T19:22:04Z (GMT). No. of bitstreams: 1 Leticia Antonelo Campos.pdf: 2246502 bytes, checksum: 0d9288acf39d871d8aedcd4d1fc4e5ca (MD5) Previous issue date: 2014-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The biofilm accumulation in the subgingival region produces an inflammatory process that can lead to alveolar bone resorption and periodontal attached loss through due inflammatory mediators that induce the production and activation of enzymes and periodontal degradation tissues. Phosphatidylserine liposomes can mimic apoptotic cells effects and may modulate the host inflammatory response. This present study evaluated the effect of phosphatidylserine liposomes on the inflammatory response and alveolar bone loss ligatureinduced in rats. We used a phosphatidylserine liposome solution preparation in order to evaluate the cytotoxicity in lineage osteoblastic cells. The antiinflammatory effect was evaluated by mouse ear edema (topical application) and rat paw edema (systemic application) methods. Then, we evaluated the phosphatidylserine liposome action on gingival inflammatory response and alveolar bone loss ligature-induced in rats. The results showed phosphatidylserine liposome 50 and 250 μM concentrations were safer considering cell viability and proliferation. The phosphatidylserine liposome gel (topical application) had no anti-inflammatory action on mouse ear edema model and alveolar bone loss ligature-induced in rats. The phosphatidylserine liposome solution (systemic application) has anti-inflammatory action on rat paw edema. Although, we did not observe an effect on alveolar bone loss ligatureinduced. We conclude phosphatidylserine liposome gel and solution are easy to prepare and they have low cost. Phosphatidylserine liposoma solution (systemic application) have antiinflammatory effect on rat paw edema model. On the other hand, it have no effect on alveolar bone loss ligature-induced in rat. / O acúmulo de biofilme na região subgengival produz um processo inflamatório que pode conduzir à reabsorção óssea e perda de inserção, devido mediadores inflamatórios que induzem a produção e ativação de enzimas e degradação dos tecidos periodontais. Lipossomas de fosfatidilserina mimetizam o efeito de células apoptóticas podendo modular a resposta inflamatória do hospedeiro. Este estudo avaliou o efeito do tratamento com lipossomas de fosfatidilserina sobre a reposta inflamatória e perda óssea alveolar induzida por ligadura em ratos. Utilizamos uma preparação de solução de lipossomas de fosfatidilserina avaliando a citotoxicidade em uma linhagem de células osteoblásticas. Avaliamos a ação anti-inflamatória por meio dos métodos de edema de orelha em camundongos (aplicação tópica) e edema de pata em ratos (aplicação sistêmica). Em seguida, avaliamos a ação de lipossomas de fosfatidilserinana na resposta inflamatória gengival e perda óssea alveolar induzida por ligadura em ratos. Os resultados mostraram que as concentrações de 50 e 250 μM dos lipossomas de fosfatidilserina foram seguras, considerando a viabilidade e proliferação celular. O gel lipossomal de fosfatidilserina (aplicação tópica) não teve ação anti-inflamatória no modelo de edema de orelha em camundongo nem na perda óssea induzida por ligadura em ratos. A solução de lipossomas de fosfatidilserina (aplicação sistêmica) teve ação anti-inflamatória no modelo de edema de pata em ratos. Porém, não observamos efeito na perda óssea induzida por ligadura. Pode-se concluir que o gel e a solução de lipossomas de fosfatidilserina são de fácil preparo e de baixo custo. Lipossomas de fosfatidilserina em solução (aplicação sistêmica) possui efeito anti-inflamatório no modelo edema de pata em ratos. Por outro lado, não interferiu na perda óssea alveolar induzida por ligadura em ratos.