Lab 7 – Bone Formation and Joints

A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)

Bone Formation and Joints

1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction Bone Formation and Joints

II. Learning Objectives
III. Slides and Micrographs 1. Bone is a specialized type of connective tissue with
A. Bone (cont.) a calcified (mineralized) extracellular matrix (ECM);
1. General structure it serves to support the body, protect internal
2. Cells organs, and acts as the body’s calcium reservoir.
a. Osteoblasts
2. Major cells of bone include: osteoblasts (form
b. Osteoclasts
osteoid which allows matrix mineralization to
B. Bone Formation occur), osteocytes (from osteoblasts; enclosed in
1. Intramembranous ossification lacunae and maintain the matrix), and osteoclasts
2. Endochondral ossification (locally erode bone matrix during bone formation
C. Joints and remodeling).
1. Synovial
3. Bone growth occurs via two basic mechanisms:
2. Intervertebral
intramembranous ossification (bone forms within
IV. Summary
mesenchymal membrane) and endochondral
ossification (bone replaces hyaline cartilage)

4. Joints are places where bones meet (articulate),

allowing at least the potential of bending or
movement; examples include, synovial joints
(diarthrosis) and intervertebral joints
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction Learning Objectives

II. Learning Objectives
III. Slides and Micrographs
1. Understand the differences and similarities between intramembranous and
A. Bone (cont.)
1. General structure
endochondral bone formation and the key function of the periosteum in
2. Cells bone growth.
a. Osteoblasts
b. Osteoclasts 2. Understand the organization of the epiphyseal growth plate and its role in
B. Bone Formation
endochondral bone formation and growth of long bones.
1. Intramembranous ossification
2. Endochondral ossification
C. Joints 3. Understand the structure of a typical synovial joint, including the nature
1. Synovial and functions of the synovium.
2. Intervertebral
IV. Summary
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I.
II.
Introduction
Learning Objectives
Slide 129: Tooth, H&E
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells 104: Bone, H&E
a. Osteoblasts
Bone
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction Slide 129: Tooth, H&E

II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
perforating
1. Intramembranous ossification
(Volkmann’s)
2. Endochondral ossification
canals
osteon
C. Joints
1. Synovial
2. Intervertebral
IV. Summary
central
canal
osteocyte
in lacuna
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015 Slide 104: Bone, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation resorption
1. Intramembranous ossification canals
2. Endochondral ossification
C. Joints
osteon
1. Synovial
2. Intervertebral
IV. Summary
central
canal

Osteon formation: resorption canals (with dimension of new osteon) are carved out of bone by
osteoclasts; blood vessels and connective tissue invade and occupy the tunnel; osteoblasts begin
to deposit new bone along the walls, forming lamellae; synthesis continues from periphery to
center until only central canal with neurovascular bundle remains in center
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015
Slide 34: Healing Bone Fracture, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
Callus
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation 1 Cortical
1. Intramembranous ossification
2. Endochondral ossification
bone
C. Joints
1. Synovial Fracture
2. Intervertebral
IV. Summary
EC
2 Hyaline
Oss
Cartilage

Sections (1) and (2) are two rib segments from a fetal/newborn rabbit; (1) gives an example of a bone
fracture and repair processes (the callus is a temporary formation of highly proliferative fibroblasts and
chondroblasts extending from the periosteum down into the fracture to form new bone); (2) gives an
example of endochondral ossification (EC Oss) bone growth
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015 Slide 34: Healing Bone Fracture, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells CT
a. Osteoblasts
b. Osteoclasts
P
B. Bone Formation Act P
1. Intramembranous ossification
2. Endochondral ossification
C. Joints M HC
1. Synovial
2. Intervertebral
Wb
IV. Summary

Tb

From the outside: (CT) is connective tissue proper surrounding the bone, note the abundant vasculature
present; (P) is periosteum with clearly visible collagen bundles and (Act P) is an area of more active
periosteum with lots of cellular differentiation occurring; (M) is marrow with abundant blood cells; (HC)
are areas of hyaline cartilage; (Wb) is woven or primary bone being deposited upon hyaline cartilage
(basophilic); there is an irregular arrangement of collagen fibers (acidophilic), large cell number, and
reduced mineral content; (Tb) is mature (mineralized), trabecular bone; close examination shows
endosteum on the edge of the bone bordering the marrow
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015
Slide 34: Healing Bone Fracture, H&E
I. Introduction CT P
II. Learning Objectives Cb
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
M
a. Osteoblasts
b. Osteoclasts
Tb
B. Bone Formation
1. Intramembranous ossification Wb
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
P
IV. Summary CT

From the outside (top): (CT) is connective tissue proper surrounding the bone; (P) is periosteum with
clearly visible collagen bundles; (Cb) is cortical bone with lamellar arrangement, note the adjacent central
canal with surrounding rings of cartilage; (M) is marrow with abundant blood cells; (Tb) is trabecular
bone, distinguished from cortical bone by the lack of osteons; close examination shows endosteum on the
edge of the bone bordering the marrow; (Wb) is woven or primary bone with an irregular arrangement of
collagen fibers, large cell number, and reduced mineral content; osteoid (unmineralized bone) is the pale,
acellular layer just below osteoblasts which line the edges of the nearby marrow cavities
 Slide 34: Healing Bone Fracture, H&E
Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts marrow
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral endosteum
IV. Summary lining
trabecula

Endosteum lines all internal surfaces of bone (both cortical and trabecular); it
is generally only a single cell‐layer thick, and consists of inactive and active
osteoblasts
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction Slide 34: Healing Bone Fracture, H&E

II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification endosteum
C. Joints
1. Synovial
2. Intervertebral
osteoid
IV. Summary lighter stained
area between
endosteum and
mature bone

mature bone
Osteoid is collagen‐rich, non‐mineralized precursor to true bone matrix; it
is secreted by osteoblasts during bone growth, repair, and remodeling;
osteoblast subsequently calcify the osteoid into bony hard bony matrix; in
the process, they become trapped in the matrix (in lacunae) and become
osteocytes
 Lab 7 – Bone Formation and Joints

Slide 34: Healing Bone Fracture, H&E

A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
inactive
1. Synovial
periosteum:
2. Intervertebral
IV. Summary
w/ flattened
osteoprogenitor
cells

maturing
bone with
osteocytes
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015
Slide 34: Healing Bone Fracture, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts active
B. Bone Formation periosteum:
1. Intramembranous ossification w/ rounded
2. Endochondral ossification or cuboidal
C. Joints osteoblasts
1. Synovial
2. Intervertebral
IV. Summary

osteocyte

Osteoblasts develop from osteoprogenitor cells (from mesenchymal cells) and are found lining the
external and internal bone surfaces; they make osteoid (collagen‐rich matrix) which they then
calcify into true hard, bony matrix; they become trapped in the matrix and become osteocytes
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I.
II.
Introduction
Learning Objectives
Slide 130: Membranous Bone, Fetal Skull
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
osteoblasts
IV. Summary

periosteum
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction
Slide 34: Healing Bone Fracture, H&E
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.) How many
1. General structure nuclei does this
2. Cells osteoclast have?
a. Osteoblasts
up to 200 is
b. Osteoclasts
possible, 5‐20 is
B. Bone Formation
usual
1. Intramembranous ossification
2. Endochondral ossification Osteoclast
C. Joints
1. Synovial Howship’s lacuna
2. Intervertebral (space)
IV. Summary

Osteoclasts are large, multinucleated cells; they resorb bone by secreting organic acids, which
dissolve hydroxyapatite, and lysosomal enzymes, which break down the osteoid matrix; at the bone
surface, osteoclasts lie in Howship's lacunae, surface depressions caused by the resorption of bone
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction
Slide 34: Healing Bone Fracture, H&E
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification Osteoclast with
C. Joints
ruffled border
1. Synovial
(visible in EM) in
2. Intervertebral
IV. Summary
Howship’s lacuna
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015
Slide 130: Membranous Bone, Fetal Skull
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral osteoclasts
IV. Summary
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction
II.
III.
Learning Objectives
Slides and Micrographs Intramembranous Ossification
A. Bone (cont.)
1. General structure
2. Cells 1. Primary center of ossification: mesenchyme cells  osteoblasts
a. Osteoblasts
b. Osteoclasts 2. Osteoblast begin depositing bony matrix to form trabeculae,
B. Bone Formation extending radially from the ossification center
1. Intramembranous ossification
2. Endochondral ossification 3. Marrow develops in spaces between trabeculae
C. Joints
1. Synovial 4. Periosteum and endosteum develop from mesenchyme
2. Intervertebral membrane
IV. Summary
5. Surfaces are remodeled to form compact bone
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015 Slide 130: Membranous Bone, Fetal Skull
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
nasal cavity with
2. Cells cartilaginous nasal
a. Osteoblasts septum in the
b. Osteoclasts middle; surrounded
by intramembranous
B. Bone Formation bone development of
1. Intramembranous ossification the skull
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral tongue in the
oral cavity
IV. Summary

developing teeth
Side View Frontal View with surrounding
intramembranous
bone development
of the jaw

Slide 130 – Fetal Skull

 Lab 7 – Bone Formation and Joints

Slide 130: Membranous Bone, Fetal Skull

A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification woven bone
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary
 Lab 7 – Bone Formation and Joints

Slide 130: Membranous Bone, Fetal Skull

A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
woven bone
1. Intramembranous ossification
2. Endochondral ossification
C. Joints hyaline
1. Synovial cartilage
2. Intervertebral
IV. Summary

periosteum
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction
Endochondral Ossification
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.) 1. Zone of Reserve Cartilage: hyaline cartilage acts as source of
1. General structure cartilage to undergo ossification
2. Cells
a. Osteoblasts 2. Zone of Proliferation: normal chondrocytes multiply
b. Osteoclasts
B. Bone Formation 3. Zone of Hypertrophy: chondrocytes enlarge and align
1. Intramembranous ossification
2. Endochondral ossification
4. Zone of Calcification: cartilage matrix calcifies; scaffold for new
C. Joints bone
1. Synovial
2. Intervertebral
5. Zone of Ossification and Resorption: chondrocytes deteriorate
IV. Summary and die; osteoblast invade and bone is deposited on the calcified
matrix; osteoclasts begin remodeling process

Because the rates of proliferation and destruction are approximately

equal, the epiphyseal plate does not change thickness; instead, it is
displaced away from the middle of the diaphysis, resulting in
growth in length of the bone
 Lab 7 – Bone Formation and Joints

I. Introduction
A560 – Fall 2015
Slide 34: Healing Bone Fracture, H&E
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification endochondral
C. Joints ossification
1. Synovial
2. Intervertebral
IV. Summary

Epiphysis Diaphysis
(End) (Middle)
Growth plate
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015 Slide 34: Healing Bone Fracture, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary

zone of zone of zone of zone of

reserve chondroblast chondrocyte calcification,
cartilage proliferation hypertrophy ossification, and
resorption
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015
Slide 34: Healing Bone Fracture, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary

zone of reserve cartilage

typical hyaline cartilage with chondrocytes arranged in small
clusters, usually only one cell per lacuna (not grouped)
 Lab 7 – Bone Formation and Joints

Slide 34: Healing Bone Fracture, H&E

A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary

zone of chondroblast proliferation

chondrocytes are dividing and increasing in number; chondrocytes are slightly larger
in size, are closer to neighboring cells, and are beginning to form rows or stacks
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015 Slide 34: Healing Bone Fracture, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary

zone of chondrocyte hypertrophy

chondrocytes are aligned in rows (along axis of growth) and are significantly
larger than in preceding zones
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015
Slide 34: Healing Bone Fracture, H&E
I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary

zone of calcification, ossification, and resorption

chondrocytes undergo apoptosis (so nuclei are more condensed); small blood
vessel and osteoprogenitor cells begin to invade, giving rise to osteoblasts
which begin to lay down bone and become osteocytes
 Fig 8‐19 Synovial Joint
Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.)
1. General structure
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
1. Intramembranous ossification
2. Endochondral ossification
C. Joints
1. Synovial
2. Intervertebral
IV. Summary

See: Fig 8‐19, 8‐20, 8‐21

 Fig 8‐22 Intervertebral Disc
Lab 7 – Bone Formation and Joints
A560 – Fall 2015

I. Introduction
II. Learning Objectives
III. Slides and Micrographs
A. Bone (cont.) Vertebra with bone
1. General structure marrow cavity (BM)
2. Cells
a. Osteoblasts
b. Osteoclasts
B. Bone Formation
Concentric layers of fibrocartilage form the
1. Intramembranous ossification
2. Endochondral ossification
annulus fibrosus (AF) (Lt. “fibrous ring”)
C. Joints
1. Synovial Nucleus pulposus (NP) is the inner core of
2. Intervertebral the vertebral disc; it is composed of a gel‐like
IV. Summary matrix consisting of water and a loose
network of collagen fibers

Vertebra with bone

marrow cavity
 Lab 7 – Bone Formation and Joints Common Confusion:
A560 – Fall 2015 Cartilage vs. Bone
I. Introduction
II. Learning Objectives
Cartilage: a specialized type of solid connective tissue
III. Slides and Micrographs which, along with bone, is distinguished by its relative
A. Bone (cont.) rigidity of the extracellular matrix (ECM); it is
1. General structure distinguishable from bone by its avascularity in addition to
its lack of lymphatics and nerves; hyaline, the most
2. Cells
common type, is pictured here
a. Osteoblasts
b. Osteoclasts Look for: (1) lack of vasculature, lymphatics, and nerves;
B. Bone Formation (2) large proportion of cells to ECM; (3) chondrocytes may
occur as pairs within a lacuna and tend to be clustered
1. Intramembranous ossification together (isogenous groups); (4) chondrocytes are usually
2. Endochondral ossification larger and rounder than osteocytes; (5) matrix is often not
C. Joints Hyaline cartilage uniform and tends to heterogeneously stain
1. Synovial
2. Intervertebral Bone: a specialized type of solid connective tissue
IV. Summary characterized by a mineralized ECM that stores calcium
and phosphate

Look for: (1) vasculature; note the central canal (osteon)

containing visible erythrocytes; (2) more ECM and fewer
cells; (3) osteocytes are singular and spaced apart; (4)
osteocytes are smaller and more condensed than
chondrocytes; (5) canals and lamellar organization may be
visible; (6) canaliculi may be visible linking osteocytes

Decalcified bone
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015 Summary
I. Introduction 1. During bone formation, woven bone (haphazard arrangement of collagen
II. Learning Objectives
fibers) gets remodeled into lamellar bone (parallel bundles of collagen in
III. Slides and Micrographs
layers known as lamellae).
A. Bone (cont.)
1. General structure
2. Periosteum is a layer of dense connective tissue on the outer surface of
2. Cells
bone; endosteum is a thin layer (generally only one cell layer) which lines
a. Osteoblasts
b. Osteoclasts
all the internal surfaces of bone.
B. Bone Formation
1. Intramembranous ossification
3. Major cells of bone include: osteoblasts (from osteoprogenitor cells; form
2. Endochondral ossification osteoid which allows matrix mineralization to occur), osteocytes (from
C. Joints osteoblasts; enclosed in lacunae and maintain the matrix), and osteoclasts
1. Synovial (from hematopoietic lineage; locally erode bone matrix during bone
2. Intervertebral formation and remodeling).
IV. Summary
4. Bone growth occurs via two basic mechanisms:
• intramembranous ossification occurs when bone forms within
mesenchymal membrane; forms bones of skull and jaw; primarily
occurs only during development or fracture repair
• endochondral ossification occurs when bone replaces hyaline
cartilage; forms and grows all other bones except as noted for IM;
occurs during development and throughout life
 Lab 7 – Bone Formation and Joints
A560 – Fall 2015 Summary (cont.)
I. Introduction 5. During epiphyseal growth (elongation of bone), the growth plate, with its
II. Learning Objectives
zonal organization of endochondral ossification, allows bone to lengthen
III. Slides and Micrographs
without the epiphyseal growth plate enlarging; zones include:
A. Bone (cont.)
• Zone of reserve cartilage
1. General structure
• Zone of proliferation
2. Cells
• Zone of hypertrophy
a. Osteoblasts
b. Osteoclasts
• Zone of calcification
B. Bone Formation
• Zone of ossification and resorption
1. Intramembranous ossification
2. Endochondral ossification 6. Joints are places where bones meet (articulate), allowing at least the
C. Joints potential of bending or movement; examples include, synovial joints
1. Synovial (diarthrosis) and intervertebral joints (with tough outer layer of
2. Intervertebral fibrocartilage known as annulus fibrosus, and gel‐like core known as
IV. Summary nucleus pulposus).
 Lab 7: Summary Features of Major Cells of Bone Tissue

Osteoblasts Osteocytes Osteoclasts

Precursor cell

Location

Percentage of all
cells in bone

Function

Appearance

Sketch
 Lab 7: Comparison of Mechanisms of Bone Formation

Intramembranous Ossification Endochondral Ossification

Bones produced

Cartilage present

Type of bone
produced

When occurs

Steps Involved