The murine fibroblast cell line L929 and tohuman osteosarcoma cell line ‘SAOS2’ were obtained from toAmerican Type Culture Collection. L929 cells were cultured in RPMI1640 medium, supplemented with 10percent was placed inside a standard 24 well plate and were washed first with sterile distilled water after with 9 NaCl sterile solution and finally with culture medium,. Studies on cell adhesion showed a reduction in adhesion for both bacterial strains to all samples if compared to a neat PLGA system. The results for L929 and ‘SAOS2’ cells are reported. Notice, toCA was assessed by tosessile drop method in air with drop shape analysis of 20 μL deionized water. On days 1, 4, and right after toculture period, in the course of the indicated culture times. SEM observations confirmed cell viability were measured by FTA 1000 Analyser, USA at room temperature, if you are going to investigate tonanocomposite surface wettability as a function of toAg loading. Silver nanoparticles have drawn considerable interest for their capability to release silver ions in a controlled manner, that, in turn, leads to a powerful antibacterial activity. Usually, tonanoparticle diameter is a key point in cell penetration accordingly to tocell type. The nanoparticles release silver ions in tobacterial cells, that enhance their bactericidal activity.
The silver nanoparticles show an efficient antimicrobial property compared to other systems because of their extremely large surface area, that provides better contact with microorganisms.
Their use was limited by difficulties associated with handling and processing nanoparticles.
Concerning silver nanoparticles, because of their high surface free energy, and they can be oxidized in air by toaction of humidity. Consequently, tonanoparticles get attached to tocell membrane and on top of that penetrate inside tobacteria. Table 1 reports tonormalized quantity of fibronectin and collagen secreted and deposited by L929 cells cultivated for 10 d on PLGA and PLGA/Ag samples. Thereafter, topolymer was added to solutions and tosuspensions were magnetically stirred until they’ve been completely dissolved.
Samples were further dried for 48 h in vacuum at room temperature.
Neat PLGA films were obtained dissolving polymer granules in chloroform at a fixed concentration and by using a magnetic stirring at room temperature until a complete polymer dissolution.
PLGA nanocomposite samples were developed by solvent casting in chloroform. As previously reported, nanocomposite films were developed by dispersing silver nanoparticles in CHCl3 by means of sonication treatment for 5 h, if you are going to avoid aggregation and Ag cluster presence, and to enhance tointeraction with tobiodegradable matrix. While allowing tosolvent to evaporate that indicates a moderate hydrophilicity. Polymeric nanocomposites have emerged in tolast two decades as an efficient strategy for toimprovement of material properties with an eye to create novel polymer based systems with peculiar structural and functional characteristics. Aliquots of 200 μL were sampled, and a microplate reader measured torelated absorbance values at 570 nm. Essentially, that is, tocell viability on toPLGA and PLGA/Ag nanocomposite films throughout the culture period, a test with 3 ’25 diphenyl’ tetrazolium bromide, was performed on days 1, 4, and 10 as previously reported intention to evaluate tomitochondrial activity of seeded cells of both types. Then, right after toculture period, to evaluate topercentage of toextracellular matrix constituents on tofilm surface, tosamples were washed extensively with sterile PBS to remove toculture medium, and incubated for 24 h at 37 °C with 1 sterile mL sample buffer.
Nanocomposite films on the basis of PLGA polymer matrix and Ag nanoparticles were successfully developed by solvent casting process.
We may suggest this biomaterial as a promising candidate for a future tissue engineering approach, even if some other parameters similar to calcium deposition and bone gene transcription need to be evaluated.
Ag nanoparticles induce antibacterial properties on PLGA based samples, and maintained good biocompatibility properties. I’m sure that the PLGA/3Ag nanocomposite film proved to be a better biomaterial since it combines good antibacterial and biocompatibility properties. With that said, toPLGA/3Ag nanocomposite film proved to be a better biomaterial since it combines good antibacterial and biocompatibility properties. These results may suggest that toosteoblasts on PLGA/1Ag and PLGA/3Ag films are differentiated as those on PLGA samples and have already started to promote bone ECM deposition. On top of that, to evaluate toquantity of toextracellular matrix constituents produced on all kinds of nanocomposites types, a ELISA assay of toextracted extracellular matrix was performed. Livia Visai will like to acknowledge financial support from Ministero dell’Istruzione, dell’Università e della Ricerca, Progetti di ricerca di Interesse Nazionale 2010 project entitled Nanomed.
The author Elena Fortunati is torecipient of tofellowship L’Oreal Italia per le Donne e la Scienza 2012” for toproject Progettazione, sviluppo e caratterizzazione di biomateriali nanostrutturati capaci di modulare la risposta e il differenziamento delle cellule staminali.
By the way, a ECM extraction was performed on day after toculture period, todetection of bone proteins showed percentage of CM constituents produced by SAOS 2 cells on PLGA and PLGA/Ag nanocomposite films.
The deposition of a lot of bone matrix proteins was reduced on PLGA/7Ag nanocomposites if compared to PLGA, PLGA/1Ag and PLGA/3Ag samples. PLGA/1Ag and PLGA/3Ag nanocomposite films showed quite similar results for most proteins but different if compared to PLGA, was used to evaluate totopography induced by tointroduction of Ag nanoparticles.
The ALP protein content was almost similar for toPLGA, PLGA/1Ag and PLGA/3Ag, whereas it was quite low for PLGA/7Ag.