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Importance of Strain Direction in Regulating Human Fibroblast Proliferation and Cytokine Secretion: A Useful in Vitro Model for Soft Tissue Injury and Manual Medicine Treatments

      Abstract

      Objective

      Manual medicine treatments (MMTs) rely on biophysical techniques that use manually guided forces in numerous strain directions to treat injuries and somatic dysfunctions. Although clinical outcomes post-MMT are positive, the underlying cellular mechanisms responsible remain elusive. We previously described an in vitro model of strain-induced tissue injury and MMTs. Using this model, the current study sought to determine if strain direction (equibiaxial [EQUI] vs heterobiaxial [HETERO]) differentially regulates human fibroblast function.

      Methods

      Fibroblasts were strained EQUI at 10% beyond their resting length for 48 hours followed by assessment of cell morphology, proliferation, and cytokine secretion via protein cytokine array and enzyme-linked immunosorbent assay (ELISA). These observations were then compared with those obtained previously for HETERO fibroblasts.

      Results

      No alterations in cell morphology were seen in EQUI fibroblasts despite our report of such changes in HETERO cells. Fibroblasts secretion profiles for 60 cytokines (via cytokine protein array) showed that in EQUI strained cells, fractalkine significantly increased (121%), whereas macrophage-derived chemoattractant/chemokine and pulmonary and activation-regulated chemokine significantly decreased (32% and 10%, respectively) compared with nonstrained cells (P < .05). The EQUI fibroblasts when compared with HETERO fibroblasts exhibited a significant decrease in proliferation (22%), inflammatory interleukin 6 secretion (75%, measured by ELISA), and macrophage-derived chemoattractant/chemokine secretion (177%, measured by ELISA, P < .05).

      Conclusions

      These divergent observations in HETERO vs EQUI strained fibroblasts may underlie the relative efficacies of MMTs carried out in different tissue strain directions. We are currently modeling MMTs such as myofascial release to further investigate this.

      Key Indexing Terms

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      References

        • Dodd JG
        • Good MM
        • Nguyen TL
        • Grigg AI
        • Batia LM
        • Standley PR
        In vitro biophysical strain model for understanding mechanisms of osteopathic manipulative treatment.
        J Am Osteopath Assoc. 2006; 106: 157-166
        • Neidlinger-Wilke C
        • Grood ES
        • Wang JHC
        • Brand RA
        • Claes L
        Cell alignment is induced by cyclic changes in cell length: studies of cells grown in cyclically stretched substrates.
        J Orthop Res. 2001; 19: 286-293
        • Berry CC
        • Cacou C
        • Lee DA
        • Bader DL
        • Shelton JC
        Dermal fibroblasts respond to mechanical conditioning in a strain profile dependent manner.
        Biorheology. 2003; 40: 337-345
        • Maheim C
        The myofascial release manual.
        3rd ed. Slack Incorporated, New Jersey2001
      1. Ward RC Foundations for osteopathic medicine. 2nd ed. Lippincott, Williams, & Wilkins, Philadelphia (PA)2003: 819-1078
        • McXIntyre C
        Therapeutic massage: an amazing modality.
        Home Health Care Manag Pract. 2004; 16: 516-520
        • Gilbert JA
        • Weinhold PS
        • Banes AJ
        • Link GW
        • Jones GL
        Strain profiles for circular cell culture plates containing flexible surfaces employed to mechanically deform cells in vitro.
        J Biomech. 1994; 27: 1169-1177
        • Van de Geest JP
        • Di Martino ES
        • Vorp DA
        An analysis of the complete strain field within Flexercell membranes.
        J Biomech. 2004; 37: 1923-1928
        • Brown TD
        Techniques for mechanical stimulation of cells in vitro: a review.
        J Biomech. 2000; 33: 3-14
        • Dixon WJ
        Processing data for outliers.
        Biometrics. 1953; 9: 74-89
        • Walters CM
        • Glucksberg MR
        • Lautenschlager EP
        • et al.
        A system to impose prescribed homogenous strains on cultured cells.
        J Appl Physiol. 2001; 91: 1600-1610
        • Lim IJ
        • Phan TT
        • Bay BH
        • et al.
        Fibroblasts cocultured with keloid keratinocytes: normal fibroblasts secrete collagen in keloid manner.
        Am J Physiol. 2002; 283: C212-C222
        • Schleip R
        • Klingler W
        • Lehmann-Horn F
        Active fascial contractility: fascia may be able to contract in a smooth muscle-like manner and thereby influence musculoskeletal dynamics.
        Med Hypotheses. 2005; 65: 273-277
        • Arora PD
        • Bibby KJ
        • McCulloch CA
        Slow oscillation of free intracellular calcium ion concentration in human fibroblasts responding to mechanical stretch.
        J Cell Physiol. 1994; 161: 187-200
        • Ko KS
        • Arora PD
        • Bhide V
        • Chen A
        • McCulloch CA
        Cell-cell adhesion in human fibroblasts requires calcium signaling.
        J Cell Sci. 2001; 114: 1155-1167
        • Mohanty MJ
        • Li X
        Stretch-induced Ca2+ release via an IP3-insensitive Ca2+ channel.
        Am J Physiol Cell Physiol. 2002; 283: C456-C462
        • Wang Y
        • Botvinick EL
        • Zhao Y
        • et al.
        Visualizing the mechanical activation of Src.
        Nature. 2005; 434: 1040-1045
        • Standley PR
        • Cammarata A
        • Nolan BP
        • Purgason CT
        • Stanley M
        Cyclic stretch induces vascular smooth muscle cell alignment via nitric oxide signaling.
        Am J Physiol. 2002; 283: H1907-H1914
        • Wang JH
        Substrate deformation determines actin cytoskeleton reorganization: a mathematical modeling and experimental study.
        J Theor Biol. 2000; 202: 33-41
        • Neidlinger-Wilke C
        • Grood ES
        • Claes L
        • Brand RA
        Fibroblast orientation to stretch begins within three hours.
        J Orthop Res. 2002; 20: 953-956
        • Lazopoulos KA
        • Staminovic D
        A mathematical model of cell reorientation in response to substrate stretching.
        Mol Cell Biochem. 2006; 3: 43-48
        • Yang G
        • Crawford RC
        • Wang JH
        Proliferation and collagen production of human patellar tendon fibroblasts in response to cyclic uniaxial stretching in serum-free conditions.
        J Biomech. 2004; 37: 1543-1550
        • Zeichen J
        • van Griensven M
        • Bosch U
        The proliferative response of isolated human tendon fibroblasts to cyclic biaxial mechanical strain.
        Am J Sports Med. 2000; 28: 888-892
        • Barkhausen T
        • van Griensven M
        • Zeichen J
        • Bosch U
        Modulation of cell functions of human tendon fibroblasts by different repetitive cyclic mechanical stress patterns.
        Exp Toxicol Pathol. 2003; 55: 153-158
        • Wang JH
        • Thampatty BP
        An introductory review of cell mechanobiology.
        Biomech Model Mechanobiol. 2006; 5: 1-16
        • Frank S
        • Kampfer H
        • Wetzler C
        • Pfeilschifter J
        Nitric oxide drives skin repair: novel functions of an established mediator.
        Kidney Int. 2002; 61: 882-888
        • Tian B
        • Liu J
        • Bitterman PB
        • Bache RJ
        Mechanisms of cytokine induced NO-mediated cardiac fibroblast apoptosis.
        Am J Physiol Heart Circ Physiol. 2002; 283: H1958-H1967
        • Meltzer KR
        • Standley PR
        Modeling repetitive motion strain and indirect osteopathic manipulative treatment in human fibroblasts: regulation of interleukin secretion.
        J Am Osteopath Assoc. 2007; ([in press])
        • Heinrich P
        • Castell J
        • Andus T
        Interleukin-6 and the acute phase response.
        Biochem J. 1990; 265: 621-636
        • Standiford T
        Anti-inflammatory cytokines and cytokine antagonists.
        Curr Pharm Des. 2000; 6: 633-649
      2. Hildebrand F, Pape HC, Krettek. The importance of cytokines in the posttraumatic inflammatory reaction. Unfallchirurg 2005;108:793-4, 796-803.

        • Gygi S
        • Rochon Y
        • Franza B
        • Aebersold R
        Correlation between protein and mRNA abundance in yeast.
        Mol Cell Biol. 1999; 19: 1720-1730
        • Huang RP
        Detection of multiple proteins in an antibody-based protein microarray system.
        J Immunol Methods. 2001; 255: 1-13
        • Wang C
        • Huang RP
        • Sommer M
        • et al.
        Array-based multiplexed screening and quantitation of human cytokines and chemokines.
        J Proteome Res. 2002; 1: 337-343
        • Lin Y
        • Huang R
        • Chen LP
        • et al.
        Profiling of cytokine expression by biotin-labeled–based protein arrays.
        Proteomics. 2003; 3: 1750-1757
        • Imai T
        • Chantry D
        • Raport CJ
        • et al.
        Macrophage-derived chemokine is a functional ligand for the CC chemokine receptor 4.
        J Biol Chem. 1998; 273: 1764-1768
        • Karlheinz F
        • Kammer W
        • Erhardt I
        • Brändlein S
        • Arnold S
        • Sebald W
        The two subunits of the interleukin-4 receptor mediate independent and distinct patterns of ligand endocytosis.
        Eur J Biochem. 1999; 265: 457-465
        • Kamkin A
        • Kiseleva I
        • Isenberg G
        Activation and inactivation of a non-selective cation conductance by local mechanical deformation of acutely isolated cardiac fibroblasts.
        Cardiovasc Res. 2003; 57: 793-803
        • Hornberger TA
        • Armstrong DD
        • Koh TJ
        • Burkholder TJ
        • Esser KA
        Intracellular signaling specificity in response to uniaxial vs. multiaxial stretch: implications for mechanotransduction.
        Am J Physiol Cell Physiol. 2005; 288: C185-C194
        • Lee CH
        • Shin HJ
        • Cho IH
        • et al.
        Nanofiber alignment and direction of mechanical strain affect the ECM production of human ACL fibroblast.
        Biomaterials. 2005; 26: 1261-1270