RESUMEN
The main aim of cell instructive materials is to guide in a controlled way cellular behavior by fine-tuning cell-material crosstalk. In the last decades, several efforts have been spent in elucidating the relations between material cues and cellular fate at the nanoscale and in the development of novel strategies for gaining a superior control over cellular function modulation. In this context, a particular attention has been recently paid to the role played by cellular membrane rearrangement in triggering specific molecular pathways linked to the regulation of different cellular functions. Here, we characterize the effect of linear microtopographies upon cellular behavior in three-dimensional (3D) environments, with particular focus on the relations linking cytoskeleton structuration to membrane rearrangement and internalization tuning. The performed analysis shown that, by altering the cellular adhesion processes at the micro- and nanoscale, it is possible to alter the membrane physical state and cellular internalization capability. More specifically, our findings pointed out that an increased cytoskeletal structuration influences the formation of nanoinvagination membrane process at the cell-material interface and the expression of clathrin and caveolin, two of the main proteins involved in the endocytosis regulation. Moreover, we proved that such topographies enhance the engulfment of inert polystyrene nanoparticles attached on 3D patterned surfaces. Our results could give new guidelines for the design of innovative and more efficient 3D cell culture systems usable for diagnostic, therapeutic, and tissue engineering purposes.
Asunto(s)
Materiales Biocompatibles/química , Nanoestructuras/química , Andamios del Tejido/química , Materiales Biocompatibles/metabolismo , Caveolinas/metabolismo , Adhesión Celular , Línea Celular , Membrana Celular/metabolismo , Clatrina/metabolismo , Citoesqueleto/metabolismo , Endocitosis , Humanos , Nanoestructuras/ultraestructura , Propiedades de Superficie , Ingeniería de TejidosRESUMEN
The development of innovative nano-bio-encapsulation systems continues to be an area of intense activity as the demand of improved delivery systems is constantly increasing in several fields including nanomedicine. For this purpose, an important goal is carrying out appropriate engineering of the surface of these nanocarriers to satisfy the organ target features for an effective in situ release and elucidate the mechanism of action which most of the time is neglected. Here, an oil-in-water (O/W) nanoemulsion coated with a polysaccharide layer film - i.e. a glycol chitosan modified with a thiol moiety - was used as nanocarrier to convey a promising poorly water-soluble nature based drug, curcumin. The final nanocarrier was completely bio-compatible and bio-stable. We investigated the enhancement of the effect of curcumin loaded in our system across monolayers of intestinal epithelial cells CaCo-2 in Transwell culture. Such in vitro platform resulted suitable to evaluate the functionality of the proposed nanocarrier and its adhesion towards the mucosal epithelial layer and, as applicative example, to investigate the anti-inflammatory effects exerted by the encapsulation of curcumin.
Asunto(s)
Comunicación Celular , Curcumina/farmacología , Emulsiones/química , Enterocitos/citología , Aceites/química , Agua/química , Células CACO-2 , Supervivencia Celular/efectos de los fármacos , Quitosano/química , Citocinas/metabolismo , Dispersión Dinámica de Luz , Enterocitos/efectos de los fármacos , Enterocitos/metabolismo , Humanos , Nanocápsulas/químicaRESUMEN
AIMS: Rat sarcoma virus (RAS)-induced tumorigenesis has been suggested to follow a three-stage model consisting of an initial RAS activation, senescence induction, and evasion of p53-dependent senescence checkpoints. While reactive oxygen species act as second messengers in RAS-induced senescence, they are also involved in oncogenic transformation by inducing proliferation and promoting mutations. In the current work, we investigated the role of extracellular superoxide dismutase (SOD3) in RAS-induced senescence and immortalization in vitro and in vivo. We used a mouse embryonic fibroblast (MEF) primary cell model along with immortalized and transformed human cell lines derived from papillary and anaplastic thyroid cancer. RESULTS: Based on our data, sod3 RNA interference in H-RasV12-transduced cells markedly inhibited cell growth, while sod3 over-expression in MEFs initially caused a proliferative burst followed by the activation of DNA damage checkpoints, induction of p53-p21 signal transduction, and senescence. Subsequently, sod3-transduced MEF cells developed co-operative p21-p16 down-regulation and acquired transformed cell characteristics such as increased telomerase activity, loss of contact inhibition, growth in low-nutrient conditions, and in vivo tumorigenesis. Interestingly, as previously reported with RAS, we showed a dose-dependent response to SOD3 in vitro and in vivo involving transcriptional and non-transcriptional regulatory mechanisms. INNOVATION: SOD3 may mediate H-RasV12-induced initiation of primary cell immortalization. CONCLUSIONS: Our results indicate that SOD3 influences growth signaling in primary and cancer cells downstream of the ras oncogene and could serve as a therapy target at an early tumorigenesis phase.
Asunto(s)
Transformación Celular Neoplásica , Embrión de Mamíferos/metabolismo , Superóxido Dismutasa/metabolismo , Animales , Embrión de Mamíferos/citología , Fibroblastos/citología , RatonesRESUMEN
Mesenchymal stromal cells (MSCs) are able to influence the growth abilities of transformed cells. Here, we show that papillary thyroid cancer TPC1 and HEK 293T cells interact physically with human primary bone marrow-derived MSCs followed by evanescence of MSC cytoplasm. Interestingly, transformed cells were able to connect only to apoptotic MSCs that had lost their migration ability, whereas naïve MSCs avoided the direct contact. The interaction stimulated the proliferation of the cocultured transformed cells, activated mitogen and stress signaling, and increased resistance to cytotoxins. Consistent with in vitro data, the MSC interaction stimulated transformed cells had enhanced ability to grow and metastasize in vivo. The parental control cells showed mild tumorigenicity as compared to MSC interaction stimulated cells yielding measurable tumors in 31 days and 7 days, respectively. Our coculture model system describes how adjacent transformed cells absorb stromal cells thereby leading to the stroma-driven evolution of moderately carcinogenic cells to highly aggressive metastatic cells.