Hyaluronic acid in Pluronic F-127/F-108 hydrogels for postoperative pain in arthroplasties: Influence on physico-chemical properties and structural requirements for sustained drug-release.

In this study, we reported the hyaluronic acid (HA) on supramolecular structure of Pluronic F-127 (PLF-127) and/or Pluronic F-108 (PLF-127) hydrogels, as well as their effects on release mechanisms, looking forward their application as lidocaine (LDC) drug-delivery systems in arthroplastic surgeries. We have studied the HA-micelle interaction using Dynamic Light Scattering (DLS), the micellization and sol-gel transition processes by Differential Scanning Calorimetry (DSC) and Rheology., of PL-based hydrogels and. The presence of HA provided the formation of larger micellar dimensions from ~26.0 to 42.4nm. The incorporation of HA did not change the micellization temperatures and stabilized hydrogels rheological properties (G'>G″), showing no interference on PL-thermoreversible properties. Small-Angle-X-ray Scattering (SAXS) patterns revealed that HA incorporation effects were pronounced for PLF-127 and PLF-108 systems, showing transitions from lamellar to hexagonal phase organization (HA-PLF-127) and structural changes from cubic to gyroid and/or cubic to lamellar. The HA insertion effects were also observed on drug release profiles, since lower LDC release constants (Krel=0.24-0.41mM·h-1) were observed for HA-PLF-127, that presented a hexagonal phase organization. Furthermore, the HA-PL systems presented reduced in vitro cytotoxic effects, pointed out their tendency to self-assembly and possible application as drug delivery systems.

Adhesion and morphology of fibroblastic cells cultured on different polymeric biomaterials.

Cell adhesion is influenced by the physical and chemical characteristics of the materials used as substrate for cell culturing. In this work, we evaluated the influence of the morphological and chemical characteristics of different polymeric substrates on the adhesion and morphology of fibroblastic cells. Cell growth on poly (L-lactic acid) [PLLA] membranes and poly(2-hydroxy ethyl methacrylate) [polyHEMA], poly(2-hydroxy ethyl methacrylate)-cellulose acetate [polyHEMA-CA] and poly(2-hydroxy ethyl methacrylate)-poly(methyl methacrylate-co-acrylic acid) [polyHEMA-poly(MMA-co-AA)] hydrogels of different densities and pore diameters was examined. Cells adhered preferentially to more negatively charged substrates, with polyHEMA hydrogels being more adhesive than the other substractes. The pores present in PLLA membranes did not interfere with adhesion, but the cells showed a distinctive morphology on each membrane.

Morphological aspects of cultured rat seminiferous tubules in the presence of fetal calf serum and follicle-stimulating hormone.

Culturing seminiferous tubules allows the analysis of spermatogenesis under controlled conditions. Reproducing the specific microenvironment for germ cells is a challenge taken up by various studies. The difficulty in supplementing all nutrients and the physical disturbances during isolation procedures cause degeneration of many cells in the seminiferous epithelium. We tested some culture conditions in order to preserve an acceptable morphology of cells inside the tubules. Seminiferous tubules were cultured during fifteen days in HAM F10 medium supplemented with fetal calf serum (FCS) and/or follicle-stimulating hormone (FSH). The cellular morphology was analyzed using scanning and transmission electron microscopy. During the culture period cellular degeneration occurred progressively. Morphological modifications of the Sertoli cells and germ cells such as accumulation of lipid droplets, nuclear and cytoplasmatic vacuolization and the presence of cell debris were observed. The addition of FCS activated the myoid cells causing nuclear rounding and thickening of the tubular wall. The best results were obtained with a serum-free culture medium supplemented with FSH.

Morphology of fibroblastic cells cultured on poly(HEMA-co-AA) substrates.

Fibroblastic cells in culture are characteristically elongated and grow in monolayers. This growth pattern can be modified by different factors, such as substrate interaction. It is characteristic of hydrogels made of poly(2-hydroxyethylmethacrylate) (polyHEMA) that they inhibit cellular attachment and spreading. Vero cells were cultured on porous samples of polyHEMA and the copolymer poly(HEMA-co-AA) with 7.5% (w/w) and 15% (w/w) acrylic acid. Cultures were maintained for 2 and 10 days in HAM F10 medium with 10% foetal calf serum. Hydrogel samples were processed for light microscopy and scanning electron microscopy. The round Vero cells proliferated on the hydrogels and were principally located inside the pores. Some cells were aggregated, but no extracellular matrix was found. The copolymer with 15% (w/w) acrylic acid was the most suitable substrate and should be used in future tests of morphological differentiation and induction of cellular function.

PolyHEMA and polyHEMA-poly(MMA-co-AA) as substrates for culturing Vero cells.

Poly (2-hydroxyethyl methacrylate), polyHEMA, is known to prevent cellular attachment and spreading. This hydrogel is used to culture cells not dependent on anchorage. Blending polyHEMA with a copolymer of methyl methacrylate and acrylic acid introduces negative charges to the hydrogel and improves its mechanical characteristics. PolyHEMA and the blend were tested for attachment and proliferation of Vero cells. Dense and porous samples of the hydrogels were used. Attachment assays included cellular quantification with MTT photometry and cellular morphology with the scanning electron microscopy after 2 h culture. Proliferation assays were carried out with 5 and 10 days culture. Cellular morphology included cytochemistry of resin sections and scanning electron microscope observations. Hydrogels allowed a few cells to attach and proliferate. The cells growing on the surface of hydrogels were organized in various layers and showed a differential morphology. Cells located inside the pores remained rounded. The hydrogels showed the possibility of inducing differentiated phenotypic expression.