Bone

Bone is a rigid and vascular supporting tissue. Like other supporting tissue it is also made
up of intercellular matrix and cells. The outer surface of the bone is covered by a dense
connective tissue layer called the periosteum, while the inner surface enclosing the
marrow cavity is lined by a thin connective tissue layer; the endosteum. Bone provide
support to fleshy structures, protects vital organs (brain, lungs and heart) and contains
bone marrow (myeloid tissue), where blood cells are formed. Bones provides movement
through the insertion of muscles and also serves as a reservoir of calcium, phosphate, and
other ions.

Surface Coverings

The periosteum It envelops outer surface of all bones except on articular surfaces and
sesamoid bones. It consists of an outer layer of dense connective tissue, which mainly
contains collagen fibers and fibroblasts. Bundles of periosteal collagen fibers, called
Sharpey’s fibers, runs at right angle to bone surface, penetrate the bone matrix and bind
the periosteum to bone. The inner, more cellular layer of the periosteum is composed of
fibroblast like cells called osteoprogenitor (osteogenic) cells, which divide by mitosis and
differentiate into osteoblasts. The periosteum is well vascularized and richly innervated.

The endosteum It lines all internal cavities within the bone and is composed of a single
layer of flattened osteoprogenitor cells and a very small amount of connective tissue.
Therefore, it is considerably thinner than the periosteum.
Mainly periosteum and endosteum provide nutrition to osseous tissue and
also provide new osteoblasts for repair or growth of bone. If osteogenic cells of
periosteum or endosteum are stimulated to proliferate, they differentiate into osteoblasts
at sites that are well vascularized and into chondroblasts at sites that are avascular.

Bone matrix Bone matrix contains both organic and inorganic components. If the
organic component is removed from the bone, the remaining calcified bone is extremely
brittle. If the mineral component is removed by prolonged exposure to acid and chelating
agents, bone becomes rubbery. Before calcification the matrix is called osteoid.

The inorganic (calcified) component It represents about 65% of the dry weight of bone
matrix. It mainly consists of calcium and phosphorus in the form of hydroxyapatite
crystals [Ca10(PO4)6(OH)2], but bicarbonate, citrate, magnesium, potassium, and sodium
are also present. This calcified matrix makes bone impermeable to diffusion of nutrients,
so the bone must be well vascularized. Inorganic components enable bone to resist
compression.

The organic component It is secreted by osteoblasts and represents about 35% of the dry
weight of bone matrix. It primarily consists of type I collagen (95%) fibers. Ground
substance is minimal and is composed of glycosaminoglycans such as chondroitin sulfate,
keratan sulfate, and some glycoproteins. Collagen fibers are arranged in the lamellae,
which are soon mineralized by crystals. Organic component enables bone to resist
tension.
Bone Cells

Osteoprogenitor cells These spindle-shaped cells are derived from embryonic

mesenchyme and are present in the periosteum, endosteum and the lining of Haversian
canal. These cells are capable of differentiating into osteoblasts. However, at low oxygen
tensions, these cells may change into chondrogenic cells.

Osteoblasts These cells are derived from osteoprogenitor cells and do not undergo
mitosis. These cells are present on innermost portion of the periosteum and in the lining
of endosteum. These cells are connected by numerous gap junctions, which facilitate
electrical or chemical communication between the cells. Each cell has a single, large and
euchromatic nucleus.

Osteoblasts are responsible for the synthesis of the organic

components of bone matrix. These cells also synthesize high levels of alkaline
phosphatase. Deposition of the inorganic components of bone also depends on the
presence of viable osteoblasts. When osteoblasts are actively involved in the synthesis of
matrix, they have a cuboidal to columnar shape and intensely basophilic cytoplasm (due
to large amounts of rough endoplasmic reticulum). This newly formed matrix surrounds
the osteoblast and now they are known as osteocyte. When they are inactive or quiescent
they become flattened and even their cytoplasm is less basophilic. During this process a
space is formed, which is occupied by osteocytes and their extensions and is known as
lacuna.

Osteoblasts secrete components of matrix at the cell surface,

producing a layer of non-calcified matrix, which lies in contact with older bone matrix,
called osteoid. This process of bone apposition is completed by subsequent deposition of
calcium salts. In the presence of alkaline phosphatase, osteoblasts facilitate the deposition
of calcium phosphate, thus mineralizing the osteoid.

Osteocytes These cells are derived from osteoblasts, and do not divide or secrete matrix.
These cells comprise 90% of all cells in the mature skeleton. Osteocytes are flat, almond-
shaped and contain less rough endoplasmic reticulum in comparison to osteoblasts. Each
cell has a single, small and heterochromatic nucleus. Cells lie in lacuna and each lacuna
contains only one osteocyte. The cytoplasmic processes of these cells lie in thin,
cylindrical canaliculi. The lacunae and canaliculi contain extracellular fluid derived from
blood vessels present in the Haversian canal. Processes of adjacent cells make contact
through gap junctions, and molecules are passed by these processes from cell to cell.
These cells transport materials between blood and bone, and are actively involved in the
maintenance of the surrounding matrix. Their death is followed by resorption of this
matrix. Osteocytes also play a role in controlling the extracellular concentration of
calcium and phosphate, because they are stimulated by calcitonin and inhibited by PTH.

Osteoclasts These cells are derived from cells in the bone marrow that are also
precursors of monocytes. These cells do not undergo mitosis. These cells are
multinucleated (up to 50 nuclei), irregularly-shaped giant cells and their cytoplasm is
usually acidophilic. Osteoclasts are present on internal surfaces as part of the endosteum
and on external surfaces as part of the osteogenic layer of the periosteum. In areas of
bone undergoing resorption, osteoclasts lie in depressions in the matrix, known as
Howship's lacunae. In active osteoclasts, the surface-facing bone matrix is highly folded,
forming a ruffled border. Surrounding the ruffled border is a clear zone that is devoid of
organelles, and contains micro- filaments, which help osteoclasts to maintain contact with
the bony surface, and serves to isolate the region of osteolytic activity. Osteoclasts cells
have receptors for calcitonin hormone.

Osteoclasts secrete acid, which decalcifies the surface layer of

bone. Acid hydrolases, collagenases, and other proteolytic enzymes secreted by
osteoclasts then degrade the organic component of the bone. Then residues of organic and
inorganic components are released into connective tissue capillaries.

Types of bone Bone exists in two main forms

1. Woven bone (immature or primary bone tissue) It is the first bone tissue to appear in
embryonic development, in fracture repair and other repair processes. It is usually
temporary and, is replaced in adults by lamellar bone except near the sutures of the flat
bones of the skull, in tooth sockets, and at the insertions of some tendons. Woven bones
have randomly organized bundles of collagen fibers along with a low mineral content and
a higher proportion of osteocytes than in lamellar bone.
2. Lamellar bone (secondary bone tissue) It replaces most woven bone. Bone is not a
static structure and is constantly being resorbed and reconstructed; therefore, lamellar
bone is also resorbed and reconstructed throughout life. In lamellar bone bundles of
collagen fibers are arranged in successive layers or lamellae. The fibers of one lamella
course at right angle to those of adjoining lamellae on either side of it. This arrangement
gives maximum rigidity and strength. It is better mineralized than woven bone, but
osteocytes are few. Lamellar bone is further of two types:
Compact bone/Dense bone/Cortical bone Structural unit of compact bone is osteon, or
Haversian system. In the osteon, lamellae are arranged as concentric cylinders
surrounding a central Haversian canal, which contains nerves, connective tissue and
blood vessels. Most recently formed lamella lies closest to this canal. Diameter of this
canal is highly variable, i.e. in younger osteon canal is large, while in older osteon canal
is small. In between the lamellae are lacunae, which contain osteocytes. Canaliculi
containing osteocyte cell processes interconnect osteocyte lacunae and also connect these
lacunae directly or indirectly to a surface bathed by tissue fluid. Canaliculi bring tissue
fluid, together with nutrients and oxygen, to all its osteocytes, enabling them to stay alive
in their heavily calcified stone-like environment. Haversian canals are connected with
each other by transverse or oblique Volkmann canals, which communicate with the
marrow cavity and the periosteum. Volkmann canals are not surrounded by concentric
lamellae. Haversian canal supplies the cells of the osteon, while Volkmann canals
provide communication between osteons. The long axis of an osteon is approximately
parallel to the major axis of stress. In between the osteons, interstitial lamellae are
present, which are incomplete or fragmented osteons and left from partial resorption of
old osteons during bone remodeling. The osteons and interstitial lamellae are sharply
outlined by a refractile line called the cement line, which consists of modified matrix.
Cement lines are not traversed by canaliculi. Lamellae present just beneath the
periosteum are known as outer circumferential lamellae, while lamellae present around
the marrow cavity are known as inner circumferential lamellae. Compact bone forms
diaphysis of long bones. It also form a thin layer on the external surface of short, flat and
irregular bones in which the core is made up of spongy bone.
Ground sections of dry compact bone present a picture of
lifeless matrix. Bone cells have disappeared and their lacunae contain air. By direct light,
lacunae are bright; by transmitted light dark. This is because of low refractive index of
the air. Canaliculi, like lacunae are dark by transmitted light. They appear either as hair-
lines or as dots, depending on the plane of section.

B. Cancellous bone/Spongy bone/Trabecular bone/ Medullary bone It is composed of

bony spicules, also called trabeculae, of varying shapes and sizes that are oriented along
the lines of stress. In the trabeculae lamellae run parallel and osteocyte lies in lacunae in
between the lamellae. Trabeculae branch and anastomose with one another. Osteocytes
receive nutrients through diffusion by cell processes that extend to the trabeculae surface.
In between the trabeculae there are many interconnecting spaces that are filled with bone
marrow. In the children bone marrow is red in which blood cells develop, and contain
40% water, 40% fat and 20% protein. In later stages of growth, and in the adult, when the
rate of blood cell formation decreased, red marrow slowly changes to yellow marrow.
Yellow marrow is made up mostly of fat cells (80% fat, 15% water, 5% protein). Under
the appropriate stimulus, yellow marrow can revert to red marrow. No true osteon
systems are present due to the thinness of the trabeculae. Although spaces present in the
spongy bones reduce the weight of the bone, yet provide tremendous strength. Spongy
bone mainly forms epiphysis of long bones. All short, flat and irregular bones consist of a
core of spongy bone. A thin rim around marrow cavity of the diaphysis of long bone
consists of spongy bone.

Ossification All bones are of mesodermal origin; mineralization of bone is known as

ossification. Bone develops in the embryo by two distinct processes: endochondral
ossification and intramembranous ossification. Although endochondral and
intramembranous ossification take place in different local environments, yet both
processes produce bone that appears histologically identical. Bone formation is
accompanied by bone resorption. The combination of bone formation and resorption is
known as remodeling of bone, occurs throughout life. Remodeling of bone is slower in
lamellar bone than in woven bone.

Endochondral Ossification In this process, bones develop from a cartilaginous model. It

begins in the second month of development. All long and short bones are formed by
endochondral ossification. In the early embryonic life, a cartilage model of the future
bone is laid down. This model is covered by a membrane called the perichondrium.
Midway along the shaft of this model a blood vessel penetrates the perichondrium,
causing the transformation of chondrogenic cells to osteoprogenitor cells.
Osteoprogenitor cells then differentiate into osteoblasts, which elaborate matrix deep to
the periosteum. Then osteoblasts form a subperiosteal bone collar via the
intramembranous bone formation. Once the perichondrium starts to form bone, it is
known as periosteum.
At the same time with the appearance of the bone collar the changes occur
in the cartilage in the center of the diaphysis (future shaft). In this center a primary
ossification center develops. At the center chondrocytes of the cartilage undergo
hypertrophy and their cytoplasm becomes vacuolated. Their lacunae enlarge and matrix
surrounding the chondrocytes becomes calcified by the deposition of calcium phosphate.
Because of calcification of matrix nutrients cannot reach the chondrocytes and they
undergo degenerative changes leading to their death. Then the intercellular substance
begins to degenerate leaving large cavities in the cartilage model. The blood vessels grow
along the spaces where cartilage cells were previously located and enlarge the cavities
further. Gradually, these spaces in the middle of the shaft join with each other, and the
marrow cavity is formed. As these developmental changes are occurring, the osteoblasts
of the periosteum deposit successive layers of bone on the outer surface so that the collar
thickens, becoming thickest in the diaphysis. The cartilage model continues to grow at its
ends, steadily increasing its length.
Secondary centers of ossification develop at the epiphysis in a sequence
of similar events that form the primary center, except a bone collar is not formed. After
secondary ossification centers have formed, bone tissue completely replaced cartilage.
Cartilage remains only in two regions: the articular cartilage, which persists throughout
adult life and does not contribute to bone growth in length, and the epiphyseal plate,
which connects the epiphysis to the diaphysis.
The epiphyseal plate disappears in adults, so the growth stops in
adulthood. Once the epiphysis have closed, growth in length of bones becomes
impossible; although widening may still occur. The epiphyseal plate allows the diaphysis
of the bone to increase in length until early adulthood. Growth in diameter occurs along
with growth in length. In this process, the bone lining of the marrow cavity is destroyed
so that the cavity increases in diameter. At the same time, osteoblasts from the
periosteum add new osseous tissue around the outer surface of the bone. Initially,
ossification of diaphysis and epiphysis only produces spongy bone, but later on by
reconstruction, the outer region of spongy bone is reorganized into compact bone.

Bone growth in length Epiphyseal plate of cartilage is responsible for longitudinal

growth of bone. It can be divided into 5 zones, starting from the epiphyseal side of
cartilage, as follows:
1. Resting zone – This zone consists of small chondrocytes.

2. Proliferative zone – This zone consists of rapidly dividing chondrocytes in columns

that are parallel to the long axis of the bone.

3. Hypertrophic zone – In this zone chondrocytes become larger.

4. Calcification zone- In this zone matrix calcifies giving a granular appearance, and the
cells degenerate.

5. Ossification zone- In this zone calcified cartilage is invaded by vascular, osteogenic

tissue from the diaphysis. Osteoblasts cover the remnants of calcified cartilage, and form
bone on it.

Intramembranous Ossification In this process, bones develop from condensation of

connective tissue mesenchyme. Most of the flat bones are formed by intramembranous
ossification. The frontal and parietal bones, parts of the maxilla, mandible, occipital, and
temporal bones are formed by this process. This process also contributes to the growth of
short bones and the thickening of long bones. At the sites where bone is to form, the
mesenchyme becomes richly vascularized and show active proliferation to form
osteoprogenitor cells. These osteoprogenitor cells enlarge rapidly and differentiate into
osteoblasts, which produce bone matrix. As calcification occurs, osteoblasts are
surrounded by their own matrix and known as osteocytes. Calcification of matrix does
not kill the osteocyte as they get nutrients and oxygen via canaliculi. Bone development
occurs at several sites and form irregular trabeculae (fused spicules). Fusion of the bony
trabeculae produces spongy bone. As blood vessels invade the area, other
undifferentiated mesenchymal cells give rise to the bone marrow. At the sites, where
compact bone is to be form; like outer and inner tables of skull, trabeculae get thickened
and the spaces between them are gradually obliterated. The periosteum and endosteum
develop from portions of the mesenchymal layer that do not undergo ossification. Mitotic
activity of these mesenchymal cells gives rise to osteoprogenitor cells, which undergo
cell division and form more osteoprogenitor cells or differentiate into osteoblasts within
the inner layer of the developing periosteum.