Biocompatibility of Carbon Nanoparticles in HeLa Cells is Dictated by Synthesis and Sterilization Procedures
Elisa Panzarini,
Cristian Vergallo,
Stefania Mariano,
Luciana Dini
Issue:
Volume 2, Issue 1, June 2016
Pages:
1-7
Received:
26 August 2015
Accepted:
9 September 2015
Published:
14 October 2015
Abstract: Due to their unique chemical and physical properties, carbon-based NanoMaterials (C-NMs) are largely exploited in biomedicine, i.e., cell and tissue imaging, drug delivery and tissue engineering scaffold, even if reports regarding their toxicity are still conflicting. In fact, biological effects strictly depend on the dynamic physicochemical characteristics of C-NMs, which in turn are strongly influenced by the procedures of their synthesis, and nanometrological techniques, e.g., Electron Microscopy (EM)-based analysis, are becoming the main tool for researchers to characterize nanoproducts. The aim of the present work is the study of the influence of synthesis and sterilization protocols on the size, shape, stability and biocompatibility of carbon NanoParticles (C-NPs). C-NPs were synthesized by using graphite as bulk material through an electrochemical method applying a constant voltage of 30 V and different times of synthesis. The C-NPs solution was sterilized by adopting different sterilization protocols during and/or after the synthesis. Size, shape and stability were studied by TEM and spectroscopy, while biocompatibility was tested in HeLa cells. Synthesis and sterilization procedures did not influence size, shape and stability of C-NPs, but interfered with C-NPs biocompatibility. In fact, irrespective of time of electrolysis process, the NPs show spherical shape with an average diameter of 7 nm. UV-visible spectra show typical peak of carbonaceous materials that falls at 236 nm without aggregation and sedimentation. However, when NPs obtained at 90 min of synthesis were twice autoclaved the peak shifted to 257 nm. HeLa cells were incubated with different C-NPs solutions administered at different concentrations, ranging from 8×105 to 1.6×107 C-NPs/cell, for different times (4, 24 and 48h). Cell viability was C-NPs concentration- and time of culture-dependent; interestingly, also the time of electrolysis process used during particles synthesis and procedures adopted to sterilize C-NPs solutions largely influenced cells response.
Abstract: Due to their unique chemical and physical properties, carbon-based NanoMaterials (C-NMs) are largely exploited in biomedicine, i.e., cell and tissue imaging, drug delivery and tissue engineering scaffold, even if reports regarding their toxicity are still conflicting. In fact, biological effects strictly depend on the dynamic physicochemical charac...
Show More
Quantitative Normal Force Measurements by Means of Atomic Force Microscopy Towards the Accurate and Easy Spring Constant Determination
Issue:
Volume 2, Issue 1, June 2016
Pages:
8-29
Received:
7 September 2015
Accepted:
6 October 2015
Published:
14 October 2015
Abstract: Due to its rapid popularity increase within last three decades, with particular focus on submicrometer quantitative surface’s properties imaging, atomic force microscopy (AFM) is still a subject of development and research in terms of both better understanding and efficient utilization of various measurement techniques. Quantitative and comparable measurements at nanoscale are a significant issue, as both: science and industry desire reliable results, allowing to perform repetitive experiments at any time and location. Therefore a numerous analysis and research projects were carried out to provide metrological approach for those techniques in terms of providing the traceability and the uncertainty estimation. In this paper an overview of various methods and approaches towards quantitative determination of the normal spring constant of the AFM probes is presented.
Abstract: Due to its rapid popularity increase within last three decades, with particular focus on submicrometer quantitative surface’s properties imaging, atomic force microscopy (AFM) is still a subject of development and research in terms of both better understanding and efficient utilization of various measurement techniques. Quantitative and comparable ...
Show More
Microbial Cells Force Spectroscopy by Atomic Force Microscopy: A Review
Livia Angeloni,
Daniele Passeri,
Melania Reggente,
Fabrizio Pantanella,
Diego Mantovani,
Marco Rossi
Issue:
Volume 2, Issue 1, June 2016
Pages:
30-40
Received:
18 September 2015
Accepted:
14 October 2015
Published:
15 October 2015
Abstract: Bacterial adhesion and biofilm formation are important phenomena which can produce both detrimental and beneficial effects in several fields. Research is thus focused on the modulation of the properties of material surfaces in order to design and develop substrates able to control bacterial adhesion process, which is the first trigger event of biofilm formation. Several theoretical predictions and experimental procedures have been developed to investigate the physical, chemical and biological mechanisms regulating the attachment of bacteria to solid substrates. Nevertheless, a comprehensive understanding has not been achieved yet, limiting the capability of individuating effective technological strategies to achieve the desired bacterial adhesion behavior. The development of new experimental procedures able to furnish deeper information about bacterial adhesion mechanism is thus needed. Microbial cell force spectroscopy (MCFS) is an atomic force microscopy (AFM) based technique, consisting in the detection of force-distance curves using particular probes obtained immobilizing bacterial cells at the free end of a flexible microcantilever, which allows the detection of the different kinds of cell-surface interaction forces. In this work, we review the state of the art in the development of MCFS, focusing on its working principle and applications. A brief description of the current models and conventional experimental procedures used to evaluate bacterial adhesion to surfaces is reported. Then, the instrumentation and the working principle, the current procedures used to prepare bacterial cells probes and the main applications of the technique are described with the aim of pointing out the advantages of the technique and the limits which still have to be overcome.
Abstract: Bacterial adhesion and biofilm formation are important phenomena which can produce both detrimental and beneficial effects in several fields. Research is thus focused on the modulation of the properties of material surfaces in order to design and develop substrates able to control bacterial adhesion process, which is the first trigger event of biof...
Show More
