Phys4530B**Skeletal*Health*and*

Diseases*
Frank*Beier**DSB0035A,*;eier@uwo.ca*
Cheryle*Sguin**cheryle.seguin@schulich.uwo.ca*
*Stephen*Sims**stephen.sims@schulich.uwo.ca*
*
*

Course'Schedule'
Car.lage'and'Osteoarthri.s'(5'sec.ons;'Beier)'
*1.*IntroducIon*to*the*Skeleton*and*Skeletal*Development***Jan*7*
*2.*CarIlage*Biology*and*Pathology*of*OsteoarthriIs**Jan*14*
*3.*Student*Discussions**Jan*21**
*4.*Diseases*of*Ectopic*OssicaIon,*Metabolic*Roles*of*the*Skeleton**
*
*Jan*28*
*5.*Student*Discussions**Feb*4*
*6.*Midterm*Exam**Feb*11*
*
Spine'and'Skeletal'Regenera.on'(3'sec.ons;'Sguin)'
*7.*Spine*Biology*and*DegeneraIve*Disc*Disease**Feb.*25*
*8.*Skeletal*regeneraIon**March*4*
*9.*Mechanobiology*of*the*skeleton*(Sguin)**March*11*
*
Bone'and'Osteoporosis'(3'sec.ons;'Dixon)'
*10.*Endocrinology*of*the*skeleton**March*18*
*11.*Bone*remodeling**March*25**
*12.*Osteoporosis*and*related*metabolic*bone*diseases**Apr*1*

Marks'and'Exams'
debates

10*%*parIcipaIon,*15*%*paper*summary,*25*%*
midterm*and*50*%*nal*exam*
Exams*will*be*largely*short*and*long*answer*
quesIons*
Emphasis*will*be*on*understanding*concepts*
and*connecIng*content*from*dierent*
lectures*

Musculoskeletal'Diseases'in'Canada'
1/3 canadians

More*than*11*million*Canadians*suer*from*MSK*
diseases*(4*million*arthriIs,*the*majority*osteoarthriIs;*2*
million*osteoporosis)*
In*2000,*MSK*diseases*caused*more*than*22*billion*CAN*$*
in*direct*and*indirect*costs**the*highest*amount*of*all*
disease*groups*
Number*of*aected*individuals*and*associated*costs*are*
expected*to*raise*dramaIcally*over*the*next*decades*
80-90% of that 4 million have this

This rise is due to a sedentary lifestyle and obesity. Also living longer and the aging baby boomer population.

Costs'of'Major'Diseases'in'Canada'
indirect costs = loss of productivity in the work place. Things like that.

Public*Health*Agency*of*Canada,*Economic*Burden*of*Illness*in*Canada*2000.*

Musculoskeletal'Diseases'in'Canada'
Impact*of*MSK*diseases*is*further*elevated*by*
comorbidiIes*and*novel*roles*of*the*skeleton*
comorbidity example: when you have osteoarthritis in hips, knees and elbows, you dont exercise and have higher cardiovascular disease risks.

Comorbidity*of*osteoarthriIs*and*other*diseases*
Bone*controls*stem*cell*niche*in*marrow*(e.g.*hematopoieIc*
cells)*
The*skeleton*acts*as*regulator*of*whole*body*metabolism**
and immune cells.

Beder*ways*to*prevent,*diagnose*and*treat*MSK*diseases*
are*clearly*required*(in*parIcular*for*osteoarthriIs)*
But*funding*for*research*on*MSK*diseases*is*
disproporIonate*

Musculoskeletal'Diseases'WorldFWide'
Lancet*December*2015**Global*Burden*of*Disease*Study*
*examined*global*impact*of*all*major*diseases*
MSK*diseases*aect*>*1.7*billion*people*directly*
4th*greatest*impact*on*death*and*disability*combined*
Increase*in*burden*from*MSK*disease:*45*%*over*the*last*
20*years*
Lower*back*pain*leading*cause*of*disability*worldwide,*
while*osteoarthriIs*shows*the*most*rapid*increase*

The'Skeleton'
mice are the model organism and will be talking a lot about
them.

Alizarin Red stains calcified mineralized tissue.

Alcian Blue stains cartilage and stains proteoglycans?

hdp://www.teachpe.com/images/anatomy/skeleton.jpg*

The'Skeleton'
Human*adult:*206*bones*
Formed*through*two*dierent*processes:*
Endochondral*ossicaIon*
Intramembraneous*ossicaIon*

Has a cartilage intermediate. Responsible for 80-90% of bone formation

No cartilage intermediate. E.g. the skull.

Divided*into**
Craniofacial*skeleton*(skull)*
Axial*skeleton*(ribs,*vertebrae)*
Appendicular*skeleton*(limbs)*
*
*

Neck and above

Body core

mainly talk about long bones.

e.g. femur, humorous, etc.

A'typical'long'bone'
there is cartilage at both ends and
is helpful for movement.

outside of cell
stem cells, immune cells, etc.
are located in this cavity
trabecular bone

cortical bone

metaphysis is the area inbetween

cartilage and bone.

hdp://www.bbc.co.uk/schools/gcsebitesize/pe/appliedanatomy/
2_anatomy_skeleton_rev4.shtml*

Func.on'of'the'skeleton'
LocomoIon*(together*with*muscles,*tendons*
etc)*
Support*of*body*funcIon,*form*(e.g.*lungs)*
ProtecIon*of*internal*organs*(brain,*heart*etc)*
Calcium,*phosphate*homeostasis*
Bone*marrow:*haematopoiesis*
Metabolic*control*
Specialized*funcIons*(e.g.*middle*ear*bones)*

Skeletal'cell'types'
Chondrocytes**carIlage*cells*
Osteoblasts**bonelforming*cells*(can*
dierenIate*further*into*osteocytes)*
Osteoclasts**bonelresorbing*cells*
Important:*communicaIon*between*dierent*
cell*types*is*criIcal*for*skeletal*development,*
growth*and*funcIon*
often disrupted in diseases related to the skeleton.

Chondrocytes'
in-between the cells are ECM

the chondrocytes create, maintain, remodel this matrix

the only cells that do this

hdp://www.scielo.org.ar/scielo.php?pid=S0327l95452005000200002&script=sci_ardext*

Osteoblasts'and'osteoclasts'

these cells fuse into the bigger one

muscles cells
also fuse.

these molecules are important for

communication

Komarova*et*al.,*Bone*2003*

Diseases'of'the'Skeleton'
Developmental*diseases*

rare diseases

GeneIc*diseases*(chondrodysplasias,*osteogenesis*
imperfecta..)*
Environmental*condiIons*(malnutriIon,*
medicaIons,*injuries..)*
cortisol is a medication we will talk about
it is for autoimmune diseases and asthma and can effect growth of bones

Diseases*in*the*adult*

common diseases

OsteoarthriIs*
Osteoporosis*
DegeneraIve*Disc*Disease/Back*Pain*

How'do'we'study'func.on'and'the'
diseases'of'the'skeleton'?'
The*skeleton*is*a*very*complex*organ*with*
mulIple*cell*types*that*interact*in*many*ways*
In*addiIon,*mechanical*factors*play*a*
prominent*role*in*controlling*skeletal*
physiology*
These*factors*cannot*be*recapitulated*
faithfully*in*vitro*
Thus,*animal*models*are*required*
which prevents use from studying just cells, cant model this complex interaction in cell culture.

we cant model normal loading

Hard to study in humans due to ethical concerns of extracting bone while alive and after death is late in disease stage.
Can have more subjects when using animals.
Long human lifespan makes studying it difficult.

The'mouse'as'model'organism'for'studies'
on'skeletal'physiology'and'disease'
Economic*model*because*of*size,*lifespan*
Possibility*for*geneIc*manipulaIon*
Transgenic,*knockout,*knockin,*condiIonal*
knockout*
understand these terms for the papers and exams

knocking out the gene in the tissue of choice instead of the whole body as we do with regular knockout.

GeneIc*modicaIons*can*be*combined*with*
other*manipulaIons*
Surgery,*diet,*drugs,*exercise*

Skeletal'Development'
Con*************

ko*************

Con*******************KO*

what happens when you take away a gene. can cause dwarfism and many
die at birth.
knockout (ko) example

shorter bones, discoloured, misshapen

Two'types'of'bone'forma.on'
Intramembraneous'ossica.on:'directly*
from*precursor*cells;*mostly*bones*of*the*
skull**
Endochondral'ossica.on:'through*a*
carIlage*intermediate;*ribs,*vertebrae,*
appendicular*skeleton*

Intramembraneous'ossica.on'

wont ask us about this process

Bradley**et*al.,*Crit.*Rev.*Eukaryote*Gene*Expr.*2011*

Bones'of'the'skull:'calvariae'

not a functional skull to protect the brain

mutation in connexions (gap junctions) (cell-cell communication)

Flenniken*et*al.,*Development*2005*

Endochondral'ossica.on'
3/4 gestation

2/3 gestation

epiphysis*

these cells are just too far way from blood vessels and undergo apoptosis
and trigger angiogenesis.

Secondary*OssicaIon*
Center*

cells at top start becoming

condrocytes

resIng*

This is when the detail starts getting real

once they become

chondrocytes, they
start producing type 2
collagen and at this
stage it is all cartilage

Growth*Plate*
Once you have secondary ossification centre, the
proliferation and hypertrophic cells are called the growth plate

proliferaIng*
hypertrophic*

Primary*OssicaIon*Center*
At the end of puberty, the chondrocytes undergo hypertrophy
and undergo apoptosis or become osteoblasts. Hence the growth
plate disappears and is replaced by bone.

SOC*

epiphysis*

Solomon*et*al.,*2008*

Chondrocyte'Dieren.a.on'

most specific marker for hypertrophic chondrocyte presence

Only made by
hypertrophic
chondrocytes

Collagen X!
BSP!
MMP13!

induces? mineralization

Cyclin D1!
helps with proliferation

breaks down cartilage matrix.

Sox9!
The major transcription
regulator of cartilage formation
Required for cartilage to form

exit the cell cycle

Mesenchymal!
precursor!
fibroblast-like cells

chondroblast!
early chondrocyte (name interchangeable)

proliferating !
chondrocyte!

hypertrophic !
chondrocyte!

The'growth'plate'

hypertrophic and proliferation

occurs all in the same column

Nilsson,*Hormone*Res.*2005*

The'growth'plate'

van*der*Eerden,*Endocr*Rev*2003*

Regula.on'of'the'growth'plate'
Endocrine*factors*(Growth*hormone,*IGF,*
thyroid*hormone,*glucocorIcoids*etc.)*
Local*growth*factors*(Hedgehog,*Wnt,*FGFs,*
BMPs,*CNP*etc.)*
NutriIon*
Biomechanical*factors*
A lot of the endocrine factors influence the local growth factors

need biomechanical stimulation for normal growth

Regula.on'of'bone'growth'by'CFtype'
natriure.c'pep.de'
example that shows that drugs can
be

malformed because it
grew too fast

C-type natriuretic peptode

Agoston*et*al.,*BMC*Dev*Biol*2007*

Regula.on'of'bone'growth'by'ac.nF
modifying'drugs'
example that shows that drugs can be
used to influence bone growth

control'

A.'Woods'

Cytochalasin'D'

Jasplakinolide'

1'mm'

Growth'disorders'
Longitudinal*growth*and*therefore*nal*height*
is*controlled*by*the*growth*plate*
Each*form*of*growth*retardaIon*therefore*
involves*the*growth*plate,*directly*or*indirectly*
GeneIc*defects*directly*aecIng*the*growth*
plates:*chondrodysplasias*
Examples*of*chondrodysplasias:*mutaIons*in*
FGFR3*(achondroplasia),*Sox9,*collagen*II**

Growth'disorders'II'
Growth*failure*can*also*be*caused*by:**
Endocrine*disorders*
MedicaIon*
Mal/undernutriIon*

Dexamethasone'suppresses'bone'growth'
Control

10-6 M

10-7 M

10-8

Average'change'in'length'
(mm)'

0.75*

0.50*

**

0.25*

0.00*

DMSO*
Control*

10l6*M**
DEX*

**

**

10l7*M**
DEX*

10l8*M**
DEX*

Treatments'
**Signicantly*dierent*with*a*plvalue*less*than*0.05*

1 mm

Baybayan,'
unpublished'

Growth'and'Ossica.on'
Growth*plate*controls*longitudinal*growth*of*
endochondral*bones*
However,*ordered*transiIon*from*hypertrophic*
carIlage*to*bone*is*equally*important*for*bone*
health*
This*involves*interacIons*of*chondrocytes,*
osteoblasts,*osteoclasts*and*endothelial*cells;*e.g.:*
apoptosis*of*chondrocytes,*blood*vessel*invasion,*
degradaIon*of*chondrocyte*matrix,*dierenIaIon*
of*osteoblasts,*modeling*of*bone*

Example:'Car.lage'controls'bone'forma.on'

Car.lage'Development'and'Osteoarthri.s'
Endochondral''
ossica.on'

chondrogenesis'

Ar.cular''
car.lage'

car.lage''
growth'
'
Repair?'
'
permanent''
car.lage'
'

car.lage''
degrada.on'
'
Osteoarthri.s'