19 - Analysis of various parameters of electric current & its effect on expression of bone markers and Osteosarcoma bone cells

Abstract

Background

The exogenous application of direct current has been widely explored in tissue engineering and has promising applications in cardiac tissues, nerve tissues, and bone regeneration due to its piezoelectric properties. Research done as early as the 1980s has demonstrated the potential of direct electric current for stimulating bone cells, however, the influence of various parameters of electric current being applied remained poorly understood.

Objective

To study the qualitative and quantitative effect of exogenous electric current (square waveform with varying pulsed frequency and amount of current) on Saos2 cells, and its impact on osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-light -chain-enhancer of activated beta cells (RANK-L).

Material and Methods

The experimental setup was divided into groups, A - 1-second pulsed frequency (SPF) and B - 10-SPF. Each experimental group was subdivided into 5 subgroups (A1-A5, B1-B5) based on the current levels (10 µA -30 µA). The reverse piezoelectricity was applied 24 hours a day for 3 days. Two µlt of media were collected at 1, 3, 5, 7, 12, 24, 48, and 72hrs. which, was subjected to an ELISA. The cells were subjected to histological examination after 72 hours. The experiments were run in triplicate. The data obtained were subjected to statistical analysis.

Results

OPG and RANK-L expression was maximum at 30 µA at 24 hours in group B and peak level of OPG was delayed in group A to 48 hours. Peak RANK-L expression for group A was seen after 24 hours at 25 µA. There is an increasing trend of OPG expression at 1, 5, 12, and 24 hrs. respectively. Qualitative histological analysis showed an increase in the number and size of Saos2 cells at 30 µA current level and 10-second pulsed frequency.

Conclusion

Bioelectric stimulation can modulate both OPG and RANK-L levels. Unique frequencies can be applied to bone cells to regulate, alter, and optimize the expression of OPG and RANK-L.