Physical labeling of papillomavirus-infected, immortal, and cancerous cervical epithelial cells reveal surface changes at immortal stage.

A significant change of surface features of malignant cervical epithelial cells compared to normal cells has been previously reported. Here, we are studying the question at which progressive stage leading to cervical cancer the surface alteration happens. A non-traditional method to identify malignant cervical epithelial cells in vitro, which is based on physical (in contrast to specific biochemical) labelling of cells with fluorescent silica micron-size beads, is used here to examine cells at progressive stages leading to cervical cancer which include normal epithelial cells, cells infected with human papillomavirus type-16 (HPV-16), cells immortalized by HPV-16, and carcinoma cells. The study shows a statistically significant (at p < 0.01) difference between both immortal and cancer cells and a group consisting of normal and infected. There is no significant difference between normal and infected cells. Immortal cells demonstrate the signal which is closer to cancer cells than to either normal or infected cells. This implies that the cell surface, surface cellular brush changes substantially when cells become immortal. Physical labeling of the cell surface represents a substantial departure from the traditional biochemical labeling methods. The results presented show the potential significance of physical properties of the cell surface for development of clinical methods for early detection of cervical cancer, even at the stage of immortalized, premalignant cells.

Cell surface as a fractal: normal and cancerous cervical cells demonstrate different fractal behavior of surface adhesion maps at the nanoscale.

Here we show that the surface of human cervical epithelial cells demonstrates substantially different fractal behavior when the cell becomes cancerous. Analyzing the adhesion maps of individual cervical cells, which were obtained using the atomic force microscopy operating in the HarmoniX mode, we found that cancerous cells demonstrate simple fractal behavior, whereas normal cells can only be approximated at best as multifractal. Tested on ~300 cells collected from 12 humans, the fractal dimensionality of cancerous cells is found to be unambiguously higher than that for normal cells.

Atomic force microscopy characterization of corneocytes: effect of moisturizer on their topology, rigidity, and friction.

BACKGROUND/PURPOSE: Atomic force microscopy (AFM) is a novel technique for skin characterization. OBJECTIVES: To develop AFM tests for characterization of the outermost epidermis layer, corneocytes. As an example, the effect of moisturizer on the corneocyte properties is studied. METHODS AND MATERIALS: Topology, rigidity, and friction (between individual corneocytes and AFM probe) of the top layer of corneocytes were measured by means of Veeco DM3100 AFM. Quench moisturizing cream was applied daily on the forearm of five volunteers for a period of 9 days. The skin flakes were collected before and after the treatment using Cuderm tape strips. No additional treatment of flakes was performed before the measurements. RESULTS: A protocol for the AFM study of corneocytes is developed. After the treatment, we observed overall smoothening of the corneocyte surface, an increase of friction, and a decrease of rigidity (the Young modulus). CONCLUSION: AFM can be used as a very sensitive tool for early detection of changes in corneocytes.

A novel in vitro stripping method to study geometry of corneocytes with fluorescent microscopy: example of aging skin.

BACKGROUND/PURPOSE: To develop modification of stripping method allowing high-resolution fluorescent visualization of corneocytes of human skin in vitro. To validate the method, the measured corneocyte areas on skin flakes are collected from individuals of different ages. MATERIALS AND METHODS: Two complimentary fluorescent dyes were used sequentially. First the adhesive layer of the stripping tape was stained with a cationic dye (rhodamine 640). This tape was used to collect skin flakes. Then both the tape and collected flakes were stained with anionic dye (fluorescein). The fluorescence of the adhesive tape exposed to the second staining is substantially decreased due to the mutual quenching of the dyes. Thirteen healthy, 6-86-year-old males participated to validate the method. The measurements were done on backhand and forearm. RESULTS: The method allows high-resolution imaging of corneocytes by means of fluorescent microscopy. Both absolute areas and the dependence of corneocyte areas on the individual age are in good agreement with the data reported previously. CONCLUSION: The developed method is fast and easy. It requires minimum interaction with the individual and allows using a broad variety of fluorescent dyes that may be potentially unsafe but beneficial for imaging. It can be used on any part of human or animal body.

Atomic force microscopy detects differences in the surface brush of normal and cancerous cells.

The atomic force microscope is broadly used to study the morphology of cells, but it can also probe the mechanics of cells. It is now known that cancerous cells may have different mechanical properties to those of normal cells, but the reasons for these differences are poorly understood. Here, we report quantitatively the differences between normal and cancerous human cervical epithelial cells by considering the brush layer on the cell surface. These brush layers, which consist mainly of microvilli, microridges and cilia, are important for interactions with the environment. Deformation force curves obtained from cells in vitro were processed according to the 'brush on soft cell model'. We found that normal cells have brushes of one length, whereas cancerous cells have mostly two brush lengths of significantly different densities. The observed differences suggest that brush layers should be taken into account when characterizing the cell surface by mechanical means.

Silica nanoparticles to polish tooth surfaces for caries prevention.

Although silica particles have been used for tooth polishing, polishing with nanosized particles has not been reported. Here we hypothesize that such polishing may protect tooth surfaces against the damage caused by cariogenic bacteria, because the bacteria can be easily removed from such polished surfaces. This was tested on human teeth ex vivo. The roughness of the polished surfaces was measured with atomic force microscopy (AFM). A considerably lower nanometer-scale roughness was obtained when silica nanoparticles were used to polish the tooth surfaces, as compared with conventional polishing pastes. Bacterial attachment to the dental surfaces was studied for Streptococcus mutans, the most abundant cariogenic bacteria. We demonstrated that it is easier to remove bacteria from areas polished with silica nanoparticles. The results demonstrate the advantage of using silica nanoparticles as abrasives for tooth polishing.