Cellular Response to Mechanical Force · 2018. 4. 2. · Transduction Transcription & Translation Mechanoreceptors we know Joint Receptors and Muscle Spindle Mechanoreceptors we know

CPTA Annual Conference September 20, 2013

Sharon L. Dunn, PT, PhD, OCS 1

Cellular Response to Mechanical Force

Sharon L. Dunn, PT, PhD, OCS

Associate Professor, LSUHSC

A Clinician’s Approach to Basic Science

• Application of practice to science

• Quest for relevance

• So much to do…….

– so little time

Genome Mapping and PT

• Genetic Profiling – Susceptibility to disease

– Specificity of management

• Genetic Engineering – Gene therapy

– Regenerative medicine

• Influence of our interventions on genetic regulation – “Mechanotransduction”

www.cottonexpressions.com

Mechanotransduction: “Our” Basic Science?

Transduction Transcription & Translation

Mechanoreceptors we know Joint Receptors and Muscle Spindle

Mechanoreceptors we know Hair Cells – Inner Ear

Oghalai lab - Stanford

Gellespie & Muller Cell, 2009

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=-pAnrHE3HZ6wlM&tbnid=JoIvIH0gWpUvFM:&ved=0CAUQjRw&url=http://www.ilo.org/safework_bookshelf/english?content&nd=857170117&ei=ipcgUqOKH4SS9gSUu4CYAQ&bvm=bv.51495398,d.eWU&psig=AFQjCNHNHOUW7cZqVsgSydtGKfuS8u3DEw&ust=1377954034474560

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=-pAnrHE3HZ6wlM&tbnid=JoIvIH0gWpUvFM:&ved=0CAUQjRw&url=http://www.ilo.org/safework_bookshelf/english?content&nd=857170117&ei=ipcgUqOKH4SS9gSUu4CYAQ&bvm=bv.51495398,d.eWU&psig=AFQjCNHNHOUW7cZqVsgSydtGKfuS8u3DEw&ust=1377954034474560



Cellular Receptors for Mechanical Loads

Zollner, et al, J Mechanical Behavior of Biomedical Materials, 2013

Cellular Receptors for Mechanical Loads:

Focal Adhesion Complex

http://www.answers.com/topic/mechanotransduction

Dr. Hislop’s Dream Wolff’s Law

German anatomist and surgeon Julius Wolff (1836–1902)

“Every change in the function of a bone is followed by certain definite changes in its internal architecture and its external conformation.”

Physical Stress Theory Tissue Adaptation to Physical Stress Cellular Mechanisms

Basic Review of Cell Biology

Cell Structure

• Membrane

• Receptors

• Cytoskeleton

• Organelles

Cell Function

• Signaling

• Transduction

• Transcription

• Translation

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=0xNrb3vW1jcUnM&tbnid=ACCQywIx-ENrmM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S1751616113001069&ei=kJ8gUoKsM4je9ASokoCwDA&psig=AFQjCNGjuFLBwtIe1tdXD8oHlkTlZmQMvw&ust=1377955875777893

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=ioRLwT9w0cgu_M&tbnid=tMk5M8eLHglVeM:&ved=0CAUQjRw&url=http://pt.usc.edu/About_The_Division/Division_History/&ei=o5EgUuGONtjH4APFvoCwAw&bvm=bv.51495398,d.cWc&psig=AFQjCNFFsXMj4lMI1nHVY02iUKG8GO2bxg&ust=1377952390211540

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=Tv81DfPp9NV9zM&tbnid=8goeKUM0U8H91M:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S0889857X05701198&ei=1bMgUpm6CdfG4AOF-YHgBg&bvm=bv.51495398,d.cWc&psig=AFQjCNFPQvxM7apNTekJAEHF3M4VpD-HCg&ust=1377961265160550

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=kuWp0K2BLky5-M&tbnid=1FyMmdo9jGUPnM:&ved=0CAUQjRw&url=http://sbrsport.wordpress.com/2013/08/06/wolffs-law-and-coming-out-winter-hybernation/&ei=V7QgUoW8IIj84APvwYCoCg&bvm=bv.51495398,d.cWc&psig=AFQjCNFPQvxM7apNTekJAEHF3M4VpD-HCg&ust=1377961265160550



Cell Structure: Membrane

• Lipid bilayer

• Integral proteins

• Lipid rafts

Cell Structure: Receptors


Cell Structure: Cytoskeleton Cell structure and motility

http://thebuildingblockofbiology.wordpress.com/

Cell Structure: Cytoskeleton

Extracellular Matrix More than just “Ground Substance”

Brizzi, et al, 2012

Tensegrity

Dalby, Medical Engineering and Physics, 2005

Ingber, MD, PhD Harvard

http://en.wikibooks.org/wiki/File:Lipid_Raft*.png

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=0xNrb3vW1jcUnM&tbnid=ACCQywIx-ENrmM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S1751616113001069&ei=kJ8gUoKsM4je9ASokoCwDA&psig=AFQjCNGjuFLBwtIe1tdXD8oHlkTlZmQMvw&ust=1377955875777893

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=KIs2aXovkJj1VM&tbnid=CtWeIFCsvfN4qM:&ved=0CAUQjRw&url=http://thebuildingblockofbiology.wordpress.com/category/friendship-of-animal-and-plant-cell-what-we-have-in-common/cytoskeleton/&ei=nNMmUrWfHNPd4AOi9YGwDg&psig=AFQjCNH8D6qT1bSNIcOEpe5VqyIuye9-bQ&ust=1378362388938475

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=azi6ehtKEgPZCM&tbnid=ktYen3Uzav2ShM:&ved=0CAUQjRw&url=http://en.wikipedia.org/wiki/Cytoskeleton&ei=i9cmUu3_IJHM9gS9_IDwDw&bvm=bv.51495398,d.cWc&psig=AFQjCNEDV7PXcsBMmD1SwxLvIpM9p0TItA&ust=1378363534664152

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=WNc7-DkSjpXVqM&tbnid=ub8VzmwHRT2ESM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S1350453305000822&ei=6uAmUr051MbgA8_cgMgI&psig=AFQjCNHrEG3gtoR71GnGamz0rSN634NDZg&ust=1378366014385419

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=fAVOTVmI1Nu-GM&tbnid=FdYXeVDV2KmL0M:&ved=0CAUQjRw&url=http://wyss.harvard.edu/viewmedia/36/sheared-tensegrity-cell-model-with-nucleus&ei=feImUq-0KtKu4APtqoCoCA&psig=AFQjCNG5kffbAZmx_cAGPujbR1b7QbH2Qw&ust=1378366263046597



Cell Structure: Organelles

http://people.eku.edu/

Cell Function: Signaling

Nature Rev: Mol Cell Biol

Molecular Biology 101

• Transduction: A signal is received and converted into an intracellular message.

• Transcription: The message is

transported to the nucleus for DNA (genetic) response - coding mRNA.

• Translation: mRNA transported

to cytosol for ribosomal decoding - 3 nucleotides (codon) of mRNA encode for an amino acid, which links up with others to form a protein.

http://sph.bu.edu/

Cell Function: Transduction

Wederell et al, Sem Cell Dev Biol, 2006 Secko, http://www.scq.ubc.ca/

DNA – Packaged in Chromosomes Unwound for Transcription

http://sph.bu.edu/

Cell Function: Transcription

http://hyperphysics.phy-astr.gsu.edu/ http://www.brooklyn.cuny.edu/

http://www.google.com/url?sa=i&source=images&cd=&cad=rja&docid=L7qBdoES1mj4tM&tbnid=zpX3TYyUp7sT9M:&ved=0CAgQjRwwAA&url=http://people.eku.edu/ritchisong/301notes1.htm&ei=E9omUt_7F-K8sQTFmIHADw&psig=AFQjCNH_G2Hgeqjxk4dVJ8T5hFj6VbmPQA&ust=1378364307439633

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=oxk-AhI83oRQoM&tbnid=M66Myz7eho0GNM:&ved=0CAUQjRw&url=http://www.nature.com/nrm/posters/signalling/green.html&ei=kb0mUv2fFc_k4AO__4DgCA&psig=AFQjCNE08YKKNyQDaUroGKgPxubv6lITeQ&ust=1378356830031875

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=BZJ3qpdvzFkoDM&tbnid=eqEFZ-P9TyrSEM:&ved=0CAUQjRw&url=http://www.scq.ubc.ca/conversing-at-the-cellular-level-an-introduction-to-signal-transduction/&ei=oLgmUsvZN9Xh4APA_oHoCA&psig=AFQjCNESQktojXkcXv69VGf4MVJJVsd6yA&ust=1378355184683246

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=UE3ir8z4_dCCyM&tbnid=8sU8Haxe3Ru-nM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S1084952106001042&ei=LcEmUq7nGZSz4AOz0IGQCw&psig=AFQjCNHCzzcas-eJwLZwQlG3D9b36J8mRQ&ust=1378357273790350

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&docid=-Esh46h8rcHdFM&tbnid=dHAe_GqRFRYUCM:&ved=0CAUQjRw&url=http://honorsmicrobiology.blogspot.com/2004_10_01_honorsmicrobiology_archive.html&ei=x7cmUt6QJ9Pe4AOi3oC4Cw&bvm=bv.51495398,d.cWc&psig=AFQjCNHjd2EOWHwFKDILeHnhmCvi413c4g&ust=1378355272048430



Cell Function: Translation

http://www.funnelbrain.com/

Cell Function: Protein Folding and Transport

Kaufman, R.J. et al. Nature Rev. Mol. Cell Biol.2002

Cellular Responses to Injury: Phases of Healing

• Inflammation

• Repair/proliferation

• Remodeling/maturation

Trauma

Rolling Leukocyte

Alon & Feigelson, Cur Opin Cell Biol 2012

Inflammatory Cascade

Nature, 420, Dec. 2002



Mast Cells

• Resident in tissue, first responders

• Release – Histamine “rubor”

– Cytokines

– Chemokines

– Proteases

– Tryptases – vasodilation – Nathan C, Nature, 420, Dec.

2002.

– Diegelmann R et al, Front in Biosc, 9, Jan. 2004.

www.pathology.duke.edu

Neutrophils

• Activated by TNF from Mast cells (or bacteria)

• Release – More cytokines

– Elastase (frees own integrins to bind ECM)

– Proteases (MMPs)

– Oxidants (activate MMPs)

• Phagocytose bacteria “puss” – Nathan C, Nature, 420, Dec. 2002.


Monocytes/Macrophages

• Activated by cytokines

• Release – TGF-ß

– PDGF

– ROI

• Phagocytose debris, damaged ECM, bacteria-filled neutrophils, etc

• Marker of phase transition from inflammatory to proliferative


www.zoology.ubc.ca

Lymphocytes

• Recruited by sustained activation of mast cells and neutrophils

• Require antigen-receptor engagement for activation

• B-lymphcytes release antibodies

• T-lymphocytes target and kill bacterial or viral cells

www.uni-mainz.de

ww

w.b

ioen

g.au

ckla

nd

.ac.

nz

Repair: Fibroblasts

• Maintain and restore structural integrity of connective tissue

• Some acquire smooth muscle features – myofibroblasts, for wound contraction

• Important for matrix production during repair

www.molbiolcell.org

Remodeling

Moore et al, Dev Cell Rev, 2010



Remodeling


Daley W P et al. J Cell Sci 2008;121:255-264

©2008 by The Company of Biologists Ltd

ECM-mediated Effects on Matrix Remodeling

Remodeling


Tissue Specific Cells: Mechanotransduction and

Genetic Response to Loading

• Bone

• Ligament

• Tendon

• Cartilage

Bone Osteocyte Transduction

Santos et al, Osteoporos Int (2009) 20:1027–1031

Burger, E. H., Klein-Nulend, J, FASEB J. 13 (Suppl.), S101–S112 (1999)

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=l6CPWUfKQskc-M&tbnid=XLO33GzhBOmCZM:&ved=0CAUQjRw&url=http://bioserv.fiu.edu/~walterm/gen_bio_II/sum10_reviewmini_skeletal_muscle_organization.htm&ei=_sAgUuLlD-3j4APGloCAAg&bvm=bv.51495398,d.cWc&psig=AFQjCNEkjwu2WvzcvUkgTvviKg6LMnqZ4A&ust=1377964660125205

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=mR9dSsymWhVT5M&tbnid=hpyHKO--mBt3EM:&ved=0CAUQjRw&url=http://www.intechopen.com/books/implant-dentistry-a-rapidly-evolving-practice/bone-biology-for-implant-dentistry-in-atrophic-alveolar-ridge-theory-and-practice&ei=EMIgUrK0Cs-u4AOckICwBA&psig=AFQjCNGU5V1Tai2sp7P7WjPsmi7-vTzVIA&ust=1377964916435301



Ligament Ligament Response to Load

Woo et al J Orthop Res, 1987

Tendon Tendon Response to Loading Design

Exercise Group Non-exercise Group

4 weeks

biologic testing

6 weeks mechanical testing

Right hind limb - experimental Left hind limb - negative control

Novel Method of Immobilization Exercise Protocol

• Rats placed under anesthesia

• Splint removed for exercise

• Passive ROM of hip, knee and ankle joints initiated 3 days p.o.

• 12 cycles per minute, 5 minutes, 3 times a week

• Splint replaced

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=zvwTFcBCP-QjTM&tbnid=v-kWcgcGcaveNM:&ved=0CAUQjRw&url=http://classes.midlandstech.edu/carterp/courses/histology/ct/sld021.htm&ei=m8IgUpugLPK54APC5YG4Bg&psig=AFQjCNHpJ_3sDQllVWM0kWWB9zme0c-VVQ&ust=1377965035220710



Tendon Response to Load

Uninvolved Limb

Exercised Limb

Splinted, Unexercised Limb

Fibroblast Transduction


Fibroblast Transduction

A mouse embryonic fibroblast plated on collagen, stained for actin (green), paxillin (magenta) and p130CAS (cyan) and captured on a spinning disk microscope. This image, highlighting stress fibers and cell-matrix adhesions, was captured by Alvin Guo of the Sawada lab

A mouse embryonic fibroblast stained for the actin cytoskeleton (red) and nucleus (blue) and captured on a spinning disk microscope. This image of stress fibers was captured by Alvin Guo of the Sawada lab

Fibroblast Transduction/Transcription

Chiquet, Matrix Biol, 2003

Fibroblast Response to Strain Articular Cartilage

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=0xNrb3vW1jcUnM&tbnid=ACCQywIx-ENrmM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S1751616113001069&ei=T8YgUozRL5K24APuh4AQ&psig=AFQjCNF-Ul96EZehZWHYqigLV2Zd2SJSQw&ust=1377965496982739

http://mbi.nus.edu.sg/wp-content/uploads/2011/11/devandcancer_sawada.jpg

http://mbi.nus.edu.sg/wp-content/uploads/2011/11/devandcancer_sawada2.jpg

http://www.google.com/url?sa=i&rct=j&q=histology articular cartilage layers&source=images&cd=&cad=rja&docid=BFwpZynBqqavXM&tbnid=el3mcADAkmQPiM:&ved=0CAUQjRw&url=http://www.lab.anhb.uwa.edu.au/mb140/CorePages/Cartilage/Cartil.htm&ei=KuodUrf8EarXsASdwoD4CQ&psig=AFQjCNFTKln29rg0EEolAqKXP6ZeicF9yA&ust=1377778580227201



Cartilage Matrix Cartilage Response to Loading

• Unloaded left hind limb

• Proteoglycan stained with Alcian blue

Behrens et al, J Orthop Res, 1989

Chondrocyte Transduction

Wong & Carter, Bone, 2003

Chondrocyte Transduction Primary Cilia

Seeger-Nukpezah & Golemis, 2012

Primary Cilia: The Mechano-sensor?

Seeger-Nukpezah & Golemis, 2012

Call to Action: Khan’s call out

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=q8kkS-bG1kO-BM&tbnid=jfgEdFRBsfxtMM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S8756328203000838&ei=DMwgUrXgN7DB4APO6YCgBA&psig=AFQjCNG9d1b_Iwx9LXv_8gExrf8kHTJocA&ust=1377967482848305

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=yWnHNhwW_PPvuM&tbnid=wFEkc8M5L8cJ3M:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S0955067412000920&ei=7s4gUvKjNtSy4AOG4IHwCw&psig=AFQjCNE-_OEgNTFt9yRTWDWP28Go99faYw&ust=1377968236583328

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=yWnHNhwW_PPvuM&tbnid=qf2kzJdq1dROjM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S0955067412000920&ei=5M0gUqDTKcT-4AOMsICYCg&psig=AFQjCNGfU4wiYktMBhS1ZUZF3dThyA3N7g&ust=1377967851938564



Call to Action: PASS Call to Action: Research Collaboration

Ambrosio & Russell, JRRD, 2010

Gene Chip Technology Call to Action: Think Biologically!

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=T5mlFxnUBMv2kM&tbnid=1yeWLw4IhpbIlM:&ved=0CAUQjRw&url=http://www.doctordisruption.com/language/think-big-write-small/&ei=7vQlUsDyIJHj4AOT4oGICw&bvm=bv.51495398,d.cWc&psig=AFQjCNGnFUihSCWmfQlrJunadg4DF63PBg&ust=1378305498749777

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Cellular Response to Mechanical Force · 2018. 4. 2. · Transduction Transcription & Translation Mechanoreceptors we know Joint Receptors and Muscle Spindle Mechanoreceptors we know