Alternating electrical fields to stimulate osteogenic cells and biomimetic calcium phosphate-coated titanium
T Kreller1, A R Boccaccini1, A Jonitz-Heincke2
1Institute of Biomaterials, Department of Materials Science and Engineering, Friedrich Alexander-University Erlangen-Nuremberg, 91058 Erlangen, Germany.
Abstract:
Biophysical stimuli such as alternating electrical fields can mimic endogenous electrical potentials and currents in natural bone. This can help to improve the healing and reconstruction of bone tissue. However, little is known about the combined influence of biomaterials and alternating electric fields on bone cells. Therefore, this study aimed to investigate the impact of both, biomaterials and alternating electric fields, on osteoblast as well as osteoclast differentiation. Initially, either RAW 264.7 or MC3T3-E1 cells were seeded on Ti6Al4V substrates as a load-bearing implant material, modified with biomimetic calcium phosphate (BCP), or uncoated as a reference. The cells were stimulated towards osteoclastic and osteoblastic differentiation via respective growth factors. The effects of BCP substrate modification on cell differentiation were examined after 7 days for RAW 264.7 and after 14 days for MC3T3-E1 cells. In a further series of tests, either RAW 264.7 or MC3T3-E1 cells were seeded on BCP-modified Ti6Al4V substrates, stimulated towards differentiation using growth factors, and further electrically stimulated via alternating electric fields of different voltages and frequencies. In parallel to the first test series RAW 264.7 and MC3T3-E1 cells were stimulated for 7 and 14 days, respectively. Cell morphology was examined via scanning electron microscopy. Cell viabilities were assessed via WST-8 assay. Electrically stimulated MC3T3-E1 cell orientation was evaluated based on fluorescence microscopy images. Marker genes were examined via qPCR. While BCP increased osteoclast-specific gene expression, it had the opposite effect on osteoblast-related genes compared to respective cells seeded on uncoated Ti6Al4V substrates. ES with different parameters showed a broad cellular response due to electrocoupling. While cell viability assessments and gene expression analyses showed clear differences between ES samples and unstimulated controls, only minor cell morphology and orientation differences were observed. Furthermore, there was no clear trend towards a dominant influence of either voltage or frequency as control parameters. Further studies were initiated to investigate the underlying intracellular mechanisms targeted by ES. This work provides an introduction to the targeted control of cellular processes using defined electric fields. The optimization of voltage and frequency could provide therapeutic windows to control specific cellular functions and potentially improve bone regeneration and remodeling processes.
