RESUMEN
AIMS: There are many obstacles to overcome in the development of new drugs for metabolic diseases, including efficacy and toxicity problems in later stages of drug development. To overcome these problems and predict efficacy and toxicity in early stages, we constructed a new model of insulin resistance in terms of communication between 3T3-L1 adipocytes and RAW264.7 macrophages by three-dimensional (3D) culture. RESULTS: In this study, results focused on the functional resemblance between 3D co-culture of adipocytes and macrophages and adipose tissue in diabetic mice. The 3D mono-culture preadipocytes showed good cell viability and induced cell differentiation to adipocytes, without cell confluence or cell-cell contact and interaction. The 3D co-cultured preadipocytes with RAW264.7 macrophages induced greater insulin resistance than two-dimensional and 3D mono-cultured adipocytes. Additionally, we demonstrated that 3D co-culture model had functional metabolic similarity to adipose tissue in diabetic mice. We utilized this 3D co-culture system to screen PPARγ antagonists that might have potential as therapeutic agents for diabetes as demonstrated by an in vivo assay. CONCLUSION: This in vitro 3D co-culture system could serve as a next-generation platform to accelerate the development of therapeutics for metabolic diseases.
Asunto(s)
Evaluación Preclínica de Medicamentos/métodos , Síndrome Metabólico/tratamiento farmacológico , Síndrome Metabólico/patología , PPAR gamma/antagonistas & inhibidores , Técnicas de Cultivo de Tejidos/métodos , Células 3T3-L1 , Adipocitos/metabolismo , Adipocitos/patología , Animales , Técnicas de Cocultivo/métodos , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/patología , Hipoglucemiantes/aislamiento & purificación , Hipoglucemiantes/uso terapéutico , Macrófagos/metabolismo , Macrófagos/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Modelos Biológicos , Células RAW 264.7 , Andamios del TejidoRESUMEN
The aim of this on-site experiment is to evaluate and compare efficiencies of currently utilized biological additives to reduce emissions of dust and bioaerosol in a confinement swine house. The mean reduction rate of total dust only after spray ranged was approximately 30% for all the treatments, compared to initial level before spraying additives which was found to reduce the initial level of total dust significantly (p < 0.05). The mean reduction rate of all the treatments at 1 hr after spraying was about 24% which was 6% lower than only after spray. Since 3 hr after spraying, however, total dust level fluctuated inconstantly for all the treatments, besides application of soybean oil. The mean reduction rates of all the treatments only after spraying as compared to initial level before spraying were about 53% for total airborne bacteria (p < 0.01) and 51% for total airborne fungi (p < 0.01), respectively. At 1 hr after spraying, the reduction rate of total airborne fungi averaged to about 35% for all the treatments (p < 0.05), while insignificant reductions of total airborne bacteria were found only in the treatments with salt water, soybean oil, artificial spice, and essential oil (p > 0.05). The fluctuations of total airborne bacteria and fungi, which were similar to total dust, were observed for all the treatments 3 hr after spray.