Background: Sound-based interventions are attracting growing scientific interest as potential complementary approaches to promoting health and well-being. Information Medicine provides harmonic sound frequencies that can be delivered through its streaming platform. In the present study, the sound files were played through a conventional external speaker, and only the effects of sound exposure were investigated. The aim of this study was to use animal-free cell biological test methods to investigate the effects of appropriate harmonic sound frequencies on the regeneration of cultured connective tissue fibroblasts.

Experimental: The Information Medicine streaming platform with an individual premium account (www.informationmedicine.org) was used in a preliminary experiment to apply the sound files to the fibroblasts at the beginning of the regeneration process for 1 to 3 × 30 min. Two sound files were used simultaneously: (1) “Connective Tissue – Fascia” and (2) “Regeneration.” The two sound files were applied to the fibroblasts using a 2-way speaker system with 28 watts RMS power and separate controls for treble and bass levels (GigaWorks T20 Series II; Creative), which was connected to a personal computer and the premium information software. Since the application of 3 × 30 min had the strongest effect, this exposure duration was used for the five subsequent experiments. Statistical analysis was performed using the two-tailed Wilcoxon-Mann-Whitney rank-sum test.

Results: The preliminary experiment examining the time-dependent response of connective tissue fibroblasts to the harmonic sound files demonstrated a statistically significant improvement in regeneration at 2 × 30 min and 3 × 30 min (p ≤ 0.01). Based on the preliminary experiment, the 3 × 30 min harmonic sound file treatment was used in the subsequent five independent experiments, which confirmed improved regeneration of connective tissue fibroblasts. Taken together, the overall improvement in connective tissue fibroblast regeneration across all five independent experiments was 26.8 ± 7.4% (mean value ± standard deviation, n = 5) compared with untreated controls. This demonstrates a statistically significant improvement (p ≤ 0.01) associated with the harmonic frequencies of the sound files used here at the cellular level.

Conclusions: The present findings demonstrate that exposure to the selected harmonic sound files significantly enhanced fibroblast gap closure under the experimental conditions used in this study. As fibroblast migration and proliferation are fundamental processes in connective tissue repair, these findings point to a biologically relevant interaction between harmonic sound exposure and cellular regenerative activity. These findings support the potential biological relevance of harmonic sound frequencies as used within the Information Medicine approach.