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. 2010 Apr;14(2):162-71.
doi: 10.1016/j.jbmt.2010.01.002. Epub 2010 Jan 29.

In Vitro Modeling of Repetitive Motion Injury and Myofascial Release

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Free PMC article

In Vitro Modeling of Repetitive Motion Injury and Myofascial Release

Kate R Meltzer et al. J Bodyw Mov Ther. .
Free PMC article

Abstract

Objective: In this study we modeled repetitive motion strain (RMS) and myofascial release (MFR) in vitro to investigate possible cellular and molecular mechanisms to potentially explain the immediate clinical outcomes associated with RMS and MFR.

Method: Cultured human fibroblasts were strained with 8h RMS, 60s MFR and combined treatment; RMS+MFR. Fibroblasts were immediately sampled upon cessation of strain and evaluated for cell morphology, cytokine secretions, proliferation, apoptosis, and potential changes to intracellular signaling molecules.

Results: RMS-induced fibroblast elongation of lameopodia, cellular decentralization, reduction of cell to cell contact and significant decreases in cell area to perimeter ratios compared to all other experimental groups (p<0.0001). Cellular proliferation indicated no change among any treatment group; however RMS resulted in a significant increase in apoptosis rate (p<0.05) along with increases in death-associated protein kinase (DAPK) and focal adhesion kinase (FAK) phosphorylation by 74% and 58% respectively, when compared to control. These responses were not observed in the MFR and RMS+MFR group. Of the 20 cytokines measured there was a significant increase in GRO secretion in the RMS+MFR group when compared to control and MFR alone.

Conclusion: Our modeled injury (RMS) appropriately displayed enhanced apoptosis activity and loss of intercellular integrity that is consistent with pro-apoptotic dapk-2 and FAK signaling. Treatment with MFR following RMS resulted in normalization in apoptotic rate and cell morphology both consistent with changes observed in dapk-2. These in vitro studies build upon the cellular evidence base needed to fully explain clinical efficacy of manual manipulative therapies.

Figures

Figures 1
Figures 1
AB. Still images captured from video clips of a clinical MFR treatment. (A) Clinician’s hands and patient’s back before treatment, and (B) the same placements during a 90 second MFR. Note the simultaneous superior, lateral and clockwise strain directions.
Figure 2
Figure 2
Strain paradigm specifics for repetitive motion strain (RMS; A), and a complete 60 second cycle of modeled myofascial release (MFR; B).
Figure 3
Figure 3
Representative photomicrographs of human fibroblast construct morphology, growth patterns and actin architecture of the four treatment groups: Control, repetitive motion strain (RMS), myofascial release (MFR), and RMS+MFR.
Figure 4
Figure 4
Cell area (top), perimeter (middle) and area:perimeter (bottom) assessed from photomicrographs via digital image capturing. Different letters denote significant relationships among groups (one-way ANOVA with post hoc Tukey Multiple Comparisons Test; p<0.05; N=42 to 48 cells analyzed per treatment group.
Figure 5
Figure 5
Proliferation indicies, as measured calorimetrically with the CellTiter 96® Aqueous One Solution cell proliferation assay, of the treatment groups as a percent of non-strain control; N=3-4 (p>0.05).
Figure 6
Figure 6
Cell counts per high powered field (HPF; N=3 to 4 experiments), DNA, protein, and protein:DNA concentrations of treatment groups as a percent of control; N=2.
Figure 7
Figure 7
Apoptosis indices of treatment groups as a percent of positive control. Significance determined via one-way ANOVA with post hoc Tukey Multiple Comparisons Test; p<0.05; N=2; *= significantly different from all other groups.

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