History of Cell Culture

From the ancient Romans, through the Middle Ages, to the late of the nineteenth century, the Aristotelian doctrine of spontaneous generation was one of the most basic laws. Even the invention of the microscope and investigations of Leeuwenhoek and Hook did not disprove the Aritostelian doctrine. Finally, in the eighteenth century, the spontaneous generation doctrine was laid by Louis Pasteur. Moreover, in the first decade of the eighteenth century, nucleus was observed in plant and animal tissues, and Virchow and other scientists presented the view that cells are formed via scission of preexisting cells. In the first decade of the twentieth century, Ross Harrison developed the first techniques of cell culture in vitro, and Burrows and Carrel improved Harrison\u27s cell cultures. In mid‐twentieth century, the basic principles for plant and animal cell cultures in vitro were developed, and human diploid cell lines were established. On the basis of knowledge about the cell cycle and gene expression regulation, the first therapeutic proteins were produced using mammalian cell cultures. The end of twentieth century and early twenty‐first century brought the progress in 3‐D cell culture technology and created the possibility of the tissue engineering and the regenerative medicine development

General Cytotoxicity and Its Application in Nanomaterial Analysis

The recent increasing interest in the use of different nanoparticles in biological and medical applications encouraged scientists to analyse their potential impact on biological systems. The biocompatibility analyses of novel materials for medical applications are conducted using quantitative and qualitative techniques collected by the International Standards Organization (ISO). The well-known assays, such as tetrazolium-based assays used for mitochondrial function monitoring, LDH for membrane permeability determination and neutral red uptake (NRU) describing lysosome function, need to be optimised due to specific properties of wide range of nanomaterials. Physicochemical properties of nanoparticles (NPs) such as size, composition, concentration, shape and surface (e.g., charge, coating, aspect ratio), as well as the cell type play a crucial role in determining the nanomaterial toxicity (also uptake pathway(s) of NPs). Different nanomaterials exhibit different cytotoxicity from relatively non-toxic hexagonal boron nitride to rutile TiO2 NPs that induce oxidative DNA damage in the absence of UV light. Finally, the results of the nanomedical analysis can be enriched by holographic microscopy that gives valuable information about the doubling time (DT), cell segmentation, track cell movement and changes in cell morphology. The results can be also completed by phenotype microarrays (PMs) and atomic force microscopy (AFM) techniques that fulfil experimental data

Few-Layered Hexagonal Boron Nitride: Functionalization, Nanocomposites, and Physicochemical and Biological Properties

Hexagonal boron nitride (h-BN) is an analogue of graphite called “white graphene.” In the structure of h-BN, B and N atoms substitute C atoms. The boron and nitrogen atoms are linked via strong B-N covalent bonds and form interlocking hexagonal rings. h-BN is used in different areas due to its interesting physical and chemical properties, e.g., in electronics as an insulator and in ceramics, resins, plastics, and paints. Therefore, boron nitride (BN) is also a popular inorganic compound in cosmetic industry (the highest BN concentration up to 25% can be found in eye shadow formulation). It is also widely used in dental cement production (for dental and orthodontic applications). Boron nitride seems to be suitable for biomedical applications; therefore, the cytotoxicity in vitro and in vivo observations of h-BN nanoplates and novel few-layered h-BN-based nanocomposites are still needed. The short-time studies confirm their low cytotoxicity and suggest that BN can be used as a novel drug delivery system; however, medical application needs additional verification in long-term studies

Cytotoxicity and genotoxicity of GO-Fe3O4 hybrid in cultured mammalian cells

The study was aimed at investigating the effect of the Fe3O4  hybrid deposited on graphene oxide (GO- Fe3O4) on the relative viability and DNA integrity. The properties of the GO-Fe3O4  hybrid were analyzed using a transmission electron microscopy (TEM), X-ray diffraction technique (XRD) and thermal gravimetric method (TGA), while the efficiency of graphene oxide covalent functionalization with iron oxide nanospheres was determined by Fourier transform infrared spectroscopy (FT-IR). L929 and MCF-7 cell lines were selected to analyze the biocompatibility of GO-Fe3O4  nanoparticles. The hybrid was tested using WST-1 and LDH leakage assays. DNA integrity was analyzed by agarose gel electrophoresis and micronucleus assay was performed to examine chromosomal damage in the exposed cell lines. The tested GO-Fe3O4  hybrid did not significantly reduce cell metabolism of L929 cells. GO-Fe3O4 hybrid particles only slightly affected the integrity of cell membranes. DNA integrity and micronucleus assays did not indicate genotoxicity of the hybrid

Modulation of Cellular Response to Different Parameters of the Rotating Magnetic Field (RMF)—An In Vitro Wound Healing Study

Since the effect of MFs (magnetic fields) on various biological systems has been studied, different results have been obtained from an insignificant effect of weak MFs on the disruption of the circadian clock system. On the other hand, magnetic fields, electromagnetic fields, or electric fields are used in medicine. The presented study was conducted to determine whether a low-frequency RMF (rotating magnetic field) with different field parameters could evoke the cellular response in vitro and is possible to modulate the cellular response. The cellular metabolic activity, ROS and Ca2+ concentration levels, wound healing assay, and gene expression analyses were conducted to evaluate the effect of RMF. It was shown that different values of magnetic induction (B) and frequency (f) of RMF evoke a different response of cells, e.g., increase in the general metabolic activity may be associated with the increasing of ROS levels. The lower intracellular Ca2+ concentration (for 50 Hz) evoked the inability of cells to wound closure. It can be stated that the subtle balance in the ROS level is crucial in the wound for the effective healing process, and it is possible to modulate the cellular response to the RMF in the context of an in vitro wound healing

Carbon nanotubes with controlled length – preparation, characterization and their cytocompatibility effects

Multiwalled carbon nanotubes (MWCNTs) have attracted huge attention due to their multifunctionality. Their unique properties allows for covalent and noncovalent modifications. The most simple method for functionalization of carbon nanotubes is their decoration with the oxygen containing moieties which can be further simultaneously functionalized for design of new class carriers for targeting and imaging. Here, we present methodology for chopping nanotubes, characterization of MWCNTs, the effect of size on the biocompatibility in culture of L929 mouse fibroblasts using WST-1, LDH and apoptosis assays. The analysis provides the optimal carbon nanotubes length and concentration which can be used for functionalization in order to minimize the effect of the secondary agglomeration when interacting with cells

Hexagonal Boron Nitride Functionalized with Au Nanoparticles—Properties and Potential Biological Applications

Hexagonal boron nitride is often referred to as white graphene. This is a 2D layered material, with a structure similar to graphene. It has gained many applications in cosmetics, dental cements, ceramics etc. Hexagonal boron nitride is also used in medicine, as a drug carrier similar as graphene or graphene oxide. Here we report that this material can be exfoliated in two steps: chemical treatment (via modified Hummers method) followed by the sonication treatment. Afterwards, the surface of the obtained material can be efficiently functionalized with gold nanoparticles. The mitochondrial activity was not affected in L929 and MCF-7 cell line cultures during 24-h incubation, whereas longer incubation (for 48, and 72 h) with this nanocomposite affected the cellular metabolism. Lysosome functionality, analyzed using the NR uptake assay, was also reduced in both cell lines. Interestingly, the rate of MCF-7 cell proliferation was reduced when exposed to h-BN loaded with gold nanoparticles. It is believed that h-BN nanocomposite with gold nanoparticles is an attractive material for cancer drug delivery and photodynamic therapy in cancer killing