Andrei P. Sommer

Biostimulatory Windows in Low-Intensity Laser Activation: Lasers, Scanners, and NASA's Light-Emitting Diode Array System

OBJECTIVE The purpose of this study was to assess and to formulate physically an irreducible set of irradiation parameters that could be relevant in the achieving reproducible light-induced effects in biological systems, both in vitro and in vivo. BACKGROUND DATA Light-tissue interaction studies focusing on the evaluation of irradiation thresholds are basic for the extensively growing applications for medical lasers and related light-emitting systems. These thresholds are of central interest in the rejuvenation of collagens, photorefractive keratectomy, and wound healing. METHODS There is ample evidence that the action of light in biological systems depends at least on two threshold parameters: the energy density and the intensity. Depending on the particular light delivery system coupled to an irradiation source, the mean energy density and the local intensity have to be determined separately using adequate experimental methods. RESULTS From the observations of different research groups and our own observations, we conclude that the threshold parameters energy density and intensity are biologically independent from each other. CONCLUSIONS This independence is of practical importance, at least for the medical application of photobiological effects achieved at low-energy density levels, accounting for the success and the failure in most of the cold laser uses since Mesters pioneering work.

670 nm Laser Light and EGCG Complementarily Reduce Amyloid-β Aggregates in Human Neuroblastoma Cells: Basis for Treatment of Alzheimer's Disease?

OBJECTIVE The aim of the present study is to present the results of in vitro experiments with possible relevance in the treatment of Alzheimers disease (AD). BACKGROUND DATA Despite intensive research efforts, there is no treatment for AD. One root cause of AD is the extra- and intracellular deposition of amyloid-beta (Aβ) fibrils in the brain. Recently, it was shown that extracellular Aβ can enter brain cells, resulting in neurotoxicity. METHODS After internalization of Aβ(42) into human neuroblastoma (SH-EP) cells, they were irradiated with moderately intense 670-nm laser light (1000 Wm(-2)) and/or treated with epigallocatechin gallate (EGCG). RESULTS In irradiated cells, Aβ(42) aggregate amounts were significantly lower than in nonirradiated cells. Likewise, in EGCG-treated cells, Aβ(42) aggregate amounts were significantly lower than in non-EGCG-treated cells. Except for the cells simultaneously laden with Aβ(42) and EGCG, there was a significant increase in cell numbers in response to laser irradiation. EGCG alone had no effect on cell proliferation. Laser irradiation significantly increased ATP levels in Aβ(42)-free cells, when compared to nonirradiated cells. Laser-induced clearance of Aβ(42) aggregates occurred at the expense of cellular ATP. CONCLUSIONS Irradiation with moderate levels of 670-nm light and EGCG supplementation complementarily reduces Aβ aggregates in SH-EP cells. Transcranial penetration of moderate levels of red to near-infrared (NIR) light has already been amply exploited in the treatment of patients with acute stroke; the blood-brain barrier (BBB) penetration of EGCG has been demonstrated in animals. We hope that our approach will inspire a practical therapy for AD.

Laser modulated transmembrane convection: Implementation in cancer chemotherapy

Transmembrane diffusion imposes fundamental limits to the uptake of cytostatic drugs executing their function intracellularly. Here, we report that transmembrane convection-a mechanism exploiting the effect of moderately intense 670nm laser light on the density and viscosity of nanoscopic interfacial water layers (IWL) in the cell-forces cancer cells to uptake high doses of cytostatic drugs in a short time. Transmembrane convection is a viable alternative to established uptake forms (i.e., it works complementary to diffusive processes) and breaks the limits imposed by diffusion. We demonstrate the potency of the method in human cervical cancer cells, HeLa, using the anticancer compounds doxorubicin (DOX), methotrexate (MTX) and epigallocatechin gallate (EGCG). The method is applicable to virtually the entire chemotherapeutic arsenal and is expected to help overcome multidrug resistance in cancer cells.

A Preliminary Investigation into Light-Modulated Replication of Nanobacteria and Heart Disease

OBJECTIVE The purpose of this preliminary study is to evaluate the effect of various wavelengths of light on nanobacteria (NB). BACKGROUND DATA NB and mitochondria use light for biological processes. NB have been described as multifunctional primordial nanovesicles with the potential to utilize solar energy for replication. NB produce slime, a process common to living bacteria. Slime release is an evolutionary important stress-dependent phenomenon increasing the survival chance of individual bacteria in a colony. In the cardiovascular system, stress-induced bacterial colony formation may lead to a deposition of plaque. METHODS Cultured NB were irradiated with NASA-LEDs at different wavelengths of light: 670, 728 and 880 nm. Light intensities were about 500k Wm(-2), and energy density was 1 x 10(4) J m(-2). RESULTS Monochromatic light clearly affected replication of NB. Maximum replication was achieved at 670 nm. CONCLUSIONS The results indicate that suitable wavelengths of light could be instrumental in elevating the vitality level of NB, preventing the production of NB-mediated slime, and simultaneously increasing the vitality level of mitochondria. The finding could stimulate the design of cooperative therapy concepts that could reduce death caused by myocardial infarcts.

Light Effect on Water Viscosity: Implication for ATP Biosynthesis.

Previous work assumed that ATP synthase, the smallest known rotary motor in nature, operates at 100% efficiency. Calculations which arrive to this result assume that the water viscosity inside mitochondria is constant and corresponds to that of bulk water. In our opinion this assumption is not satisfactory for two reasons: (1) There is evidence that the water in mitochondria prevails to 100% as interfacial water. (2) Laboratory experiments which explore the properties of interfacial water suggest viscosities which exceed those of bulk water, specifically at hydrophilic interfaces. Here, we wish to suggest a physicochemical mechanism which assumes intramitochondrial water viscosity gradients and consistently explains two cellular responses: The decrease and increase in ATP synthesis in response to reactive oxygen species and non-destructive levels of near-infrared (NIR) laser light, respectively. The mechanism is derived from the results of a new experimental method, which combines the technique of nanoindentation with the modulation of interfacial water layers by laser irradiation. Results, including the elucidation of the principle of light-induced ATP production, are expected to have broad implications in all fields of medicine.

The Top of the Biomimetic Triangle

There is increasing observational evidence indicating that crystalline interfacial water layers play a central role in evolution and biology. For instance in cellular recognition processes, in particular during first contact events, where cells decide upon survival or entering apoptosis. Understanding water layers is thus crucial in biomedical engineering, specifically in the design of biomaterials inspired by biomimetic principles. Whereas there is ample experimental evidence for crystalline interfacial water layers on surfaces in air, their subaquatic presence could not be verified directly, so far. Analysing a polarity dependent asymmetry in the surface conductivity on hydrogenated nanocrystalline diamond, we show that crystalline interfacial water layers persist subaquatically. Nanoscopic interfacial water layers with an order different from that of bulk water have been identified at room temperature on both hydrophilic and hydrophobic model surfaces — in air and subaquatically. Their generalization and systematic inclusion into the catalogue of physical and chemical determinants of biocompatibility complete the biomimetic triangle.

Extraordinary Anticancer Effect of Green Tea and Red Light

OBJECTIVE Increasing observational evidence suggests that epigallocatechin gallate--the major polyphenolic component of green tea--is instrumental in suppressing the growth of cancer cells. Therefore, methods that promise to enhance the suppressive potential of green tea have the highest clinical relevance. BACKGROUND DATA Human cervical cancer cells, HeLa, the first continuous cancer cell line, represent a mainstay model in cancer research. Green tea inhibited their growth, whereas their exposure to moderate levels of laser light resulted in an opposite effect. Both effects are individually documented in the literature. METHODS HeLa cells were supplemented with green tea, irradiated with moderately intense laser light (670 nm) for 1 min, and incubated for 52 h. RESULTS We found an extraordinary inhibition of HeLa cells by a combination of green tea and red light. We achieved an inhibition of 1,460%, compared with non-irradiated samples. CONCLUSION Our result receives clinical relevance from a recent study in which epigallocatechin gallate suppressed the growth of melanoma in vivo.

On the Social Behaviour of Cells

Polystyrene Petri dishes are in use in hundreds of thousands of laboratories world wide. Cell culture experiments performed in them provide fundamental information in a wide range of applications, including but not limited to testing novel biomaterials and pharmaceuticals, and stem cell research. These experiments cost billions of dollars per year. In this study we report on a potential deficiency of polystyrene Petri dishes, possibly caused by an increase in interfacial pH under relevant culture conditions and affecting cell performance. We conclude that cell performance on Petri dishes could be improved by improving the Petri dishes. As a spin-off of our study we postulate the concept that cancer cells and stem cells are social. It is impossible to validate this concept on the basis of the model established in this paper. However, the coherence of our insights may encourage further study and lead to the development of a qualitative improvement of cell culture devices, including Petri dishes and culture flasks, to the identification of potential strategies for chemotherapy and chemoprevention that could suppress progression of metastasis, and to the establishment of improved settings for tissue engineering and stem cell research. An immediate recommendation of our study is to use chemically and biologically inert substrates for important cell culture experiments, for example, nanocrystalline diamond.

It is Time for a Change: Petri Dishes Weaken Cells

We wish to draw the attention to a potential deficiency in the biocompatibility of polystyrene cell culture dishes which is caused by a softening of the material under relevant culture conditions. The finding confirms the central hypothesis of our previous model study. In it we assumed a local increase in pH at the interface between the hydrophilic polymer and liquid. The finding is of considerable biological interest. Polystyrene tissue culture dishes are now in use for 50 years. To the best of our knowledge their biocompatibility has never been challenged. Here we report the first experimental proof that exposure to water softens the surface of polystyrene Petri dishes. We expect that our results will stimulate the development of a new generation of cell culture devices, including Petri dishes and culture flasks, and the establishment of improved biomimetic settings for tissue engineering and stem cell research. New non-swelling biomaterials or nanocoatings designed to reduce the swelling of polymer culture dishes could improve cell performance. The need for further study is clear.

Pulsed Laser Light Forces Cancer Cells to Absorb Anticancer Drugs – The Role of Water in Nanomedicine

Abstract: Anticancer drugs executing their function intracellularly enter cancer cells via diffusive processes. Complementary to these slow processes, cells can be forced to incorporate drugs by convection – a more efficient transport process. Transmembrane convection is induced by moderately intense pulsed laser light (or light emitting diodes) changing the structure of nanoscopic water layers in cells. This is a fundamental difference with the method of photodynamic therapy. In a model system we demonstrate that a total irradiation time of one minute is sufficient to completely inhibit proliferation of cancer cells. Transmembrane convection protects healthy cells from extended chemotherapy exposure, could be exploited to overcome multidrug resistance, and is a promising new tool in a variety of therapies as well as in skin rejuvenation.