4530b: Spine Biology

Dr. Cheryle A Sguin

Assistant Professor,
Department of Physiology and Pharmacology
Associate Scientist, Lawson Health Research Institute
and Childrens Health Research Institute
cheryle.seguin@schulich.uwo.ca

The problem of back pain

Low back pain is the most
frequently reported
musculoskeletal problem in
adults
Canadians have a reported
lifetime prevalence of 84%
There are symptomatic
treatments, but no diseasemodifying therapeutics for
back pain related to disc
degeneration

Learning Objectives
1. To describe the structure and function the tissues that
compose the spine
2. To understand the biology of the intervertebral disc
What are the 3 tissue types?
What are these tissues composed of?
How does each contribute to the function of the IVD?
3. Be able to discuss the concept of tissue homeostasis.

movie shown in class:

spine has 2 roles:

1. keeps us up right
2. Protects the spine

Spine consists of stacked vertebral bones.

Need to keep them stacked, with mutlple joints and connective tissue to keep them up right.
1. ???? Other joint type (synovial)
2. Intervebretal joints: cartilaginal joints (no liquid)

http://www.spineuniverse.com/anatomy/spinal-anatomy-animation

Anatomy of the Human Spine

The human spine consists of 33 vertebrae:
7 Cervical vertebra (C1-C7)
7 Cervical vertibra

12 Thoracic vertebra (T1-T12)

12 Thoracic vertebra

5 Lumbar vertebra (L1-L5)

has a tail
has a sacral region which does fuse as well
The curvature of the spine is called: kyfosa????

5 Lumbar verebra

The shape of the spine is dependent on the biomechanical loads. specifically it is by the vertebra structure and angle.

The sacrum and coccyx are made up of 9

fused vertebrae
BONUS MARK ON EXAM: Look up # of vertebra of giraffe, human and mouse.
7 cervical vertibra for a giraffe

spinemd.com

Anatomy of the Human Spine

Each vertebra is composed of
changes in this can lead to scoliosis

Vertebral body: weightsupporting cylindrical bone

(cancellous bone). The vertebral
bodies stack up on one another to
create the vertebral column.
Spinous process: bony projection
that extends dorsal and caudal
from the junction of the laminae;
attachment point for muscles and
ligaments
Lamina: forms the walls of the
spinal canal. Has four projections
called articular processes.
holds the stack together

spinemd.com

the boney projections that come out to protect the nerves

Anatomy of the Human Spine

Each vertebra is attached to
the one above and below it
by ligaments and muscles and
is separated from the
vertebra above and below it
by an intervertebral disc.
the structure separating two vertebra

Two vertebra and the disc

between them make up a
motion segment.
spineuniverse.com

Anatomy of the Human Spine

spineuniverse.com

Facet joints: Each vertebra has two sets of facet joints. Synovial joints
the top of the bone has cartilage (similar to a knee joint structure). Any disease that can occur articular joints (e.g. knee or hip) can affect the facet joint.
has a synovium and cartilage.

One pair faces upward (superior articular facet) and one downward (inferior
articular facet).
Formed by the overlapping of the upper projection of one lamina (superior
articular process) and the lower projection (inferior articular process) of the
adjacent vertebra.
a little bit of the movement happens through the facet joint, but most of the movement occurs through the intervertebra disc.

Anatomy of the Spine

Foramen
Opening between the pedicles of the vertebrae
through which nerve roots exit off the spinal
cord.
Spinal canal
Contains the spinal cord from the brain stem to
the upper lumbar spine.
There is no spinal cord in the lower lumbar
spine. At the L1 level, the spinal cord terminates
as the conus medullaris.
A bundle of nerves called the cauda equina
(meaning horse's tail) branch off the conus
medullaris. These nerve roots are suspended in
the fluid-filled dural sac.
spine ends at the lumbar, after that is is just nerves???
Pinching of the lower nerves causes __________?

spinemd.com

The Intervertebral Disc

video

3 tissue types are important

The Intervertebral Disc

Very hydrated
in the middle of IV disc
lots of water

1) Nucleus Pulposus
- Gelatinous mass composed of
small and aggregating
protoglycans, type II collagen
high water content is regulated by proteoglycan. Has a really high negative charge, so it attracts water.
Type II collagen is loose and mesh like (very little arrangement)

2) Annulus Fibrosus
- Fibrocartilagenous structure
composed of concentric
lamellae of type I collagen

Hybrid tissue (very fibrous tissue with a bit of cartilage)

Lower water content, but much higher collagen content (meant to form a rigid structure to keep
the nucleus pulpous in place

3) Cartilage Endplate
- Thin layer of hyaline cartilage
that anchors disc to adjacent
vertebral bone
Provides nutrition? for the inververtebral disc???

Sally Roberts, Jill P.G. Urban

Mammalian Spine Development

mouse embryo

The mouse node is a

teardrop-shaped pit at
the distal tip of the early
embryo formed by cells
derived from the
primitive streak
the node is a regulator

entirely symmetrical
at this point

just a pit at the base

of embryo

Node-derived signals (Nodal) establish left-right asymmetry

Nature 397, 295-298

all the cells in the node make a single cilia.

they beat in unison sends all the nodal signal to the left side.

Cells in the left LPM that receive Nodal signaling contribute to various visceral organs, such
as the lung and heart, that develop left side-specific morphologies

Mammalian Spine Development

once the cells in the node have formed those viceral organs, they migrate
up the back towards the head and form a small tube and rod like
structure (the primitive spine).
Not all the cells for the spine come from the node, but if the node doesnt
form this, then the rest of the spine wont form.

Mammalian Spine Development

notochord is the primitive spine

condensed mesochymes forms

the annulus fibrosis

cells from the mesochime come and surround where the bone develops, the

notochord and forms both condensed and notochord gets pinched off
From: the
non-condensed mesochymes
Dis Model Mech. 2011 January; 4(1): 3141.

most of the spine is mesochyme derived, but the

______ structure comes from the notochord

The Intervertebral Disc

Annulus Fibrosus
Outer
Inner

(rings)

% wet weight

Nucleus Pulposus

H2O
Coll
collagen

PG

proteoglycans

AF

NP

AF

(middle)

In adulthood the cartilage endplate

and the disc itself normally have no
blood vessels of their own but rely on
the blood supply of adjacent tissues,
such as ligaments and vertebral body,
to transport nutrients and remove
waste products. Only the outer
portion of the disc is innervated.
NP is the largest avascularized and aneural tissue in the body.
Nutrients and waste removal dependent on diffusion to surrounding tissues.
Big issue if diffusion is cut off. leads to tissue degenerations.

The Nucleus Pulposus

A proteoglycan and water gel held
together loosely by an irregular network
of fine collagen type II and elastin fibers.
The major proteoglycan of the disc is
aggrecan, which, because of its high
anionic glycosaminoglycan content (i.e.,
chondroitin sulfate and keratan sulfate),
provides the osmotic properties needed
to resist compression.
E.g. attracts water via negative charge

Tissue homeostasis is maintained by

balanced expression of anabolic
factors (i.e. matrix proteins) and
catabolic factors (i.e. enzymes that
break down the matrix)

ADAMTS
(aggrecanase)
cleave PGs

MMPs cleave
collagens, PGs

proteoglycans are the sideways lines

if you loose these, you loose the gel like properties.

The Nucleus Pulposus

This is where disc degeneration starts. First tissue to go.

Matrix is synthesized and

maintained by specialized cell types:
(lost in the first decade of life)
Notochordal
Cells
The white cells are large vacuole cells

NP (chondrocyte-like cells)
nucleus pulposus eventually gets populated with these cells

Large notochordal cells are thought to create a different matrix than NP.
Thought that notochordal loss might be also a loss of the stem cells

Degenerative Changes

Developmental origins of NP?

Few years ago, no one knew where NPs came from.
Are the the final differentiation of notochordal cells or do the migrate in from mesochymes.

# Notochord Cells

Cell type-specific markers?

We dont know what these cells are.

Age

e.g. notochordal cells decrease and NP go up.

Nucleus Pulposus: Gene Targeting

Need for a notochord / nucleus pulposus specific marker to

make tissue-specific CRE mouse
Noto is expressed in the node
and posterior notochord from
embryonic day 7.5 to 12
Yamanaka Dev 2007

The Annulus Fibrosus

Fibrocartilagenous structure
fibroblast cells embedded
between lamellae
between the bands of blue, there are streaks of red. This red is proteoglycan.
Blue/black streaks are the collagen

Collagen fibers are arranged in concentric

layers with consecutive rings running in
alternating directions, but always with an
orientation of 65 degrees.
needs to have rigidity which is why there is this fantastic collagen structure. but needs to move as well
which is why there are also proteoglycans to attact water.

The Cartilage Endplate

The vertebral endplate consists
3 cells thick.
of hyaline cartilage weakly like
very thin.
bonded to the cortical bone of
the vertebral body, and collagen
fibers of the annulus fibrosus
and nucleus pulposus.

If there is no uncalcified caritlage matrix there (e.g. a healthy cartilage matrix) there is movement
of nutrients into IV disc and no waste removal.

annulus fibrosis also anchor through the caritlage.

Intervertebral Disc Function

The muscles and ligaments holding your spine together,
also apply a constant load to your spine.

This is all from the lumbar

The main function of the disc is

mechanical.
The disc transmits load along the spinal
column and also allows the spine to
bend and twist.
The loads on the disc arise from body
weight and muscular activity, and
change with posture.
During daily activities the disc is
subject to complex loads.
Extending or flexing the spine produces
mainly tensile and compressive stresses
on the disc, which increase in magnitude
going down the spine, due to differences
in body weight and geometry.
(to absorb load)

Intervertebral Disc Function

Compression force
from body weight and
muscle contraction
(straight arrows) raises
the pressure in the
nucleus.
Most of the load is dissapated by the NP.
This load gets spread to the annulus fibrosis which is able
to resist the load.

This, in turn, increases

the tension in the
annulus (curved
arrows).
Neumann, 2010

Intervertebral Disc Function

Water is the
main
constituent in
the disc (65 to
90% of tissue
volume)

Compressive loading forces some water from the disc effectively

increasing the aggrecan concentration and its swelling potential, and
resisting further compression.
On removal of the compressive load, disc height is restored as water is
drawn back into the tissue to restore the original equilibrium conditions.
Any parameter that decreases proteoglycan concentration or weakens
the collagen network will be detrimental to disc function.
As the water moves in and out, it is bringing in nutrients and taking away waste.

Intervertebral Disc Function

Correlation between the quantity of proteoglycan and the water content of
the matrix.
The amount of water also varies depending on the load applied to the disc,
hence differs night and day when load varies.

Water is important both to the mechanical functioning and enabling

transport of dissolved substances within the matrix.

The Paradigm of Mechanical Loading