CH8 Bone

 Provides solid support for the body

 Protects vital organs such as organs in cranial and thoracic cavities
 Encloses medullary cavities containing bone marrow where blood cells
are formed.
 Bone (or osseous) tissue serves as a reservoir for calcium, phosphate and
other ions that can be released or stored in a controlled manner to
maintain constant concentrations in body fluids.
 Form a system of levers that multiply forces generated during skeletal
muscle contraction and transform them into bodily movements.
 Mineralised tissue confers mechanical and metabolic functions to
skeleton
 Specialised connective tissue composed of calcified extracellular
material, bone matrix and three main cell types: Osteocytes, osteoblasts,
and osteoclasts
 Osteocytes: found in cavities (lacunae) between bone matrix layers
(lamellae), with cytoplasmic processes in small canaliculi that extend
into the matrix.
 Osteoblasts: growing cells which synthesise and secrete organic
components of the matrix
 Osteoclasts: giant, multinucleated cells involved in removing calcified
bone matrix and remodelling bone tissue.
Exchanges between osteocytes and blood capillaries depend on
communication through very thin cylindrical spaces of canaliculi – due to
metabolites being unable to diffuse through calcified bone matrix.
All bones are lined on their internal and external surfaces by layers of
connective tissue containing osteogenic cells – endosteum on internal surface
surrounding marrow cavity and periosteum on external surface.
As it is hard, bone cannot be sectioned routinely.
Bone matrix is softened by immersion in a decalcifying solution before paraffin
embedding or embedded in plastic after fixation and sectioned with a
specialised microtome.
Bone Cells
Osteoblasts
 Originate from mesenchymal stem cells
 Produce the organic components of bone matrix including type I
collagen fibres, proteoglycans and matricellular glycoproteins such as
osteonectin.
 Deposition of inorganic components of bone depends of its activity.
 Active osteoblasts are located exclusively at surfaces of bone matrix and
are bound by integrins – forms single layer of cuboidal cells joined by
adherent and gap junctions.
 When synthetic activity is completed, some differentiate as osteocytes
entrapped in matrix-bound lacunae – some flatten and cover matrix
surface as bone lining cells and majority undergo apoptosis.
 During processes of matrix synthesis and calcification, osteoblasts are
polarised cells with ultrastructural features denoting active protein
synthesis and secretion.
 Matrix components are secreted at cell surface n contact with existing
bone matrix, producing layer of unique collagen-rich material called
osteoid between osteoblast layer and pre-existing bone surface.
 Process of bone appositional growth is completed by subsequent
deposition of calcium salts into newly formed matrix.
 Prominent among the non-collagen proteins secreted by osteoblasts is
vitamin K-dependent polypeptide osteocalcin which together with
various glycoproteins binds Ca2+ ions and concentrates this mineral
locally.
 Release membrane-enclosed matrix vesicles rich in alkaline phosphatase
and other enzymes whose activity raises local concentrations of PO43-
ions.
 In micro-environment with high concentrations of both ions, matrix
vesicles serve as foci for formation of hydroxyapatite crystals.
Osteocytes
 During transition from osteoblasts to osteocytes, cells extend many long
dendritic processes, which also become surrounded by calcifying matrix.
 Diffusion of metabolites between osteocytes and blood vessels occur
through small amount of interstitial fluid in canaliculi between bone
matrix and osteocytes and their processes.
 Communicate with each other and nearby osteoblasts and bone
ligaments cells via gap junctions at end of their processes.
 Almond-shaped osteocytes (most abundant cells in bone) exhibit
significantly less RER, smaller Golgi complexes, and condensed nuclear
chromatin than osteoblasts
 Maintain calcified matrix, and their death is followed by rapid matrix
resorption.
 Express many different proteins, including factors with paracrine and
endocrine effects that help regulate bone remodelling.
 Extensive lacunar-canalicular network of these cells and their
communication with all other bone cells allow osteocytes to serve as
sensitive detectors of stress-or-fatigue-induced microdamage in bone
and to trigger remedial activity in osteoblasts and osteoclasts.

Osteoclasts
 Very large, motile cells with multiple nuclei which are essential for
matrix resorption during bone growth and remodelling
 Large size and multinucleated condition are due to origin from fusion
of bone marrow-derived monocytes.
 Development requires two polypeptides produced by osteoblasts:
macrophage-colony-stimulating factor (M-CSF) and the receptor
activator of nuclear factor-kB ligand (RANKL).
 In areas undergoing resorption, osteoclasts on bone surface lie within
resorption lacunae (cavities in the matrix)
 In active osteoblast membrane domain that contacts the bone forms
a circular sealing zone which binds tightly to bone matrix and
surrounds ruffled border (area with many surface projections).
  Circumferential sealing zone allows formations of specialised
microenvironment between osteoclast and the matrix in which bone
resorption occurs.
 Osteoclast pumps protons into subcellular pocket to acidify and
promote dissolution of adjacent hydroxyapatite, and releases matrix
metalloproteinases and other hydrolytic enzymes from lysosome-
related secretory vesicles for localised digestion of matrix proteins.
 Activity is controlled by local signalling factors from other bone cells
 Osteoblasts activated by parathyroid hormone produce M-CSF,
RANKL, and other factors that regulate the formation and activity of
osteoclasts.

Bone Matrix
 50% of dry weight of bone matrix is inorganic materials – calcium
hydroxyapatite is most abundant. Bicarbonate, citrate, magnesium,
potassium, and sodium ions are found. Non-crystalline calcium
phosphate is also found.
 Surface of hydroxyapatite crystals are hydrated, facilitating exchange of
ions between mineral and body fluids
 Organic matter embedded in matrix I 90% type I collagen. Also includes
small proteoglycans and multi-adhesive glycoproteins such as
osteonectin
 Calcium binding proteins and phosphatases released from cells in matrix
vesicles promote calcification of matrix.
 Association of minerals with collagen fibres during calcification provides
the hardness and resistance required for bone function.
 Decalcified bone matrix is acidophilic (due to high collagen content)

Periosteum
 Periosteum covers external surface of bone.
 Organised similar to perichondrium of cartilage, with outer fibrous layer
of dense connective tissue, containing bundled type I collagen,
fibroblasts and blood vessels.
  Perforating (Sharpey) Fibres (bundles of periosteal collagen), bind
periosteum to the bone
 Periosteal blood vessels branch and penetrate bone carrying metabolites
to and from bone cells
 Inner layer is more cellular and includes osteoblasts, bone lining cells,
and mesenchymal stem cells called osteoprogenitor cells. Have potential
to proliferate extensively and produce new osteoblasts.

Endosteum
 Covers internal surfaces of bones
 Covers small trabeculae of bone matrix that project into marrow
cavities.
 Contains osteoprogenitor cells, osteoblasts, and bone lining cells within
sparse, delicate matrix of collagen fibres.
Types of Bone
Type of Bone Histological Major Locations Synonyms
Features
Woven Bone, Irregular and Developing and Immature bone;
newly calcified random growing bones; primary bone;
arrangement of hard callus of bone bundle bone
cells and collagen; fractures
lightly calcified
Lamellar Bone, Parallel bundles of All normal regions Mature bone;
remodelled from collagen in thin of adult bone secondary bone
woven bone layers (lamellae),
with regularly
spaced cells
between; heavily
calcified
Compact bone, Parallel lamellae or Thick outer region Cortical Bone
80% of all lamellar densely packed (beneath
bone mass osteon, with periosteum) of
interstitial lamellae bones
Cancellous bone, Interconnected Inner regions of Spongy bone;
20% of all lamellar thin spicules or bones, adjacent to trabecula bone;
bone mass trabeculae covered marrow cavities medullary bone
by endosteum

 Epiphyses (bulbous ends of long bones) are composed of spongy bone

covered by thin layer of compact bone.
 Diaphysis (cylindrical part of long bone) almost totally compact bone
with thin region of cancellous bone on inner surface around central
marrow cavity.
 Flat bones that form calvaria (skullcap) have two layers of compact bone
called plates, separated by thicker layer of cancellous bone called the
diploë.
 Compact and cancellous bone show two types of organisation: mature
lamellar bone with matrix existing as discrete sheets, and woven bone,
newly formed with randomly arranged components.
Lamellar Bone
 Characterised by multiple layers of lamellae of calcified matrix
 Lamellae are organised as parallel sheets or concentrically around
central canal. Type I collagen fibres are aligned in each lamella, with
pitch of fibres orientation shifted orthogonally in successive lamellae
 High ordered organisation of collagen causes birefringence with
polarising light microscopy; alternating bright and dark layers are due to
changing orientation of collagen fibres in lamellae – adds greatly to
strength of lamellar bone
 Osteon (or Haversian system) refers to complex of concentric lamellae
(100-250μm in diameter) surrounding a central canal that contains blood
vessels, nerves, and endosteum.
 Lacunae containing osteocytes are found between successive lamellae,
are all connected by canaliculi containing the cells’ dendritic processes.
 All cells of osteon receive nutrients and oxygen from vessels in central
canal through canaliculi
 Outer boundary of each osteon is a layer called the cement line which
includes more non-collagen proteins in addition to mineral and collagen.
 Osteon is a long cylinder parallel to diaphysis. Each has 5-20 concentric
lamellae around central canal which communicates with marrow cavity
and periosteum.
 Canals communicate with each other through transverse perforating
canals (or Volkmann canals) with few concentric lamellae.
 All centric osteonic canals and perforating canals form when matrix is
laid down around areas with pre-existing blood vessels.
 Irregularly shaped Interstitial lamellae are found scattered among intact
osteons. These are lamellae remaining from osteons partially destroyed
by osteoclasts during growth and remodelling of bone.
 Compact bone also includes parallel lamellae organised as multiple
external circumferential lamellae immediately beneath periosteum and
fewer inner circumferential lamellae around marrow cavity
 Lamellae of outer and innermost areas enclose and strengthen the
middle region containing vascularised osteons.
 Bone remodelling occurs continuously throughout life.
 Remodelling resorbs parts of old osteons and produces new ones in
compact bone.
  Osteoclasts remove and form small, tunnel cavities – these are quickly
invaded by osteoprogenitor cells from endosteum or periosteum and
sprouting loops of capillaries
 Osteoblasts develop, line wall of tunnels and begin to secret osteoid in a
cyclic manner, forming a new osteon with concentric lamellae of bone
trapped osteocytes.

Woven Bone
 Non-lamellar and characterised by random disposition of type I collagen
fibres and is the first bone tissue to appear in embryonic development
and in fracture repair.
 Usually temporary and is replaced in adults by lamellar bone, except in
few places in the body e.g. sutures of calvaria and in insertions of some
tendons.
 Has a lower mineral content and a higher proportion of osteocytes than
mature lamellar bone.
 These features reflect that immature woven bone forms more quickly
but has less strength than lamellar bone

Osteogenesis
Occurs by one of two processes:
Intramembranous ossification
 Osteoblasts differentiate directly from mesenchyme and begin secreting
osteoid.
 Bone formation within condensed mesenchyme bone formation begins
in ossification centres, areas in which osteoprogenitor cells arise,
proliferate, and form incomplete layers of osteoblasts around a network
of developing capillaries.
 Osteoid secreted by osteoblasts calcifies forming small irregular areas of
woven bone with osteocytes in lacunae and canaliculi.
 Continued matrix secretion and calcification enlarges these areas and
leads to fusion of neighbouring ossification centres
 Anatomical bone forma gradually as woven bone matrix is replaced by
compact bone that encloses a region of cancellous bone with marrow
and larger blood vessels.
  Mesenchymal regions that do not undergo ossification give rise to
endosteum and periosteum of new bone

Endochondral Ossification
 Pre-existing matrix of hyaline cartilage is eroded and invaded by
osteoblasts, which then begin osteoid production
 Forms most bones of the body and is especially well studied in
developing long bones where it consists of sequence of events.
 Perichondrium becomes vascularised
 Blood vessels supply new nutrients to mesenchymal cells which
differentiate and form osteoblasts.
 Osteoblasts gather at diaphysis wall and form bone collar.
 Chondrocytes in central cavity enlarge causing the matrix to calcify.
Matrix is now impermeable which causes cell death
 Central clearing forms where cells have died and is supported by bone
collar
 Healthy chondrocytes elsewhere cause elongation
 Periosteal bud (contains artery, vein, lymphatics, and nerves) invades
cavity causing formation of spongy and delivers osteoblast and
osteoclasts
 Bone continues to elongate
 Primary ossification centre continues to enlarge
 Osteoclasts break down newly formed spongy bone
 Medullary cavity begins to form
 Cartilage is then only deposited in epiphyses
 Bony epiphyseal surface begins to form
 Secondary ossification centre appears after birth at the epiphyses
 Usually 2 in larger bone
 Articular cartilage on surface protect and lubricates joints.
 epiphyseal cartilage which connects each epiphysis to the diaphysis and
allows longitudinal bone growth