BONE CELLS

Dr. Vaskar Humagain

Resident
Orthopaedics and Trauma Surgery
NAMS
Date: 2076/01/20 (5/3/2019)
Introduction to Bone

• is a type of hard endoskeletal and

mineralized connective tissue
• They function to protect various
organs of the body , produce red and
white blood cells, store minerals,
provide structure and support for the
body, and enable mobility.
• Bone tissue is the structural
component of the bone. It is the
specialized form of connective tissue
characterized by mineralized
extracellular matrix.
Components of osseous tissue

• Bone cells are the functional entity

that form, resorb or provide other
various types of bone cells..

• Bone matrix is the ground substance

that consists of organic and
inorganic substances. It is the
supporting medium for the bone
cells.
Cytal architechture
1. Osteoblasts
2. Osteocytes
3. Osteoclasts
4. Bone lining cells
5. Osteoprogenitor
cells
 Osteoblasts
• Mononuclear cuboidal cells that are
located along the bone surface
comprising 4–6% of the total resident
bone cells.
• Derived from undifferentiated
mesenchymal stem cells
• Two major functions
• Formation of bone
• Regulation of osteoclastic activity
Differentiation of osteoblasts
• Bone morphogenetic protein (BMP)
• members of the Wingless (Wnt) pathways 
• Runt-related transcription factors 2, Distal-
less homeobox 5 (Dlx5), and osterix (Osx) for
osteoblast differentiation
• β-Catenin (intramembranous bone formation)
• PDGF and IGF
• FGF-2 and 18, microRNAs, and connexin 43
RUNx2 and osteoblasts

• Alsoknown as core-binding
factor subunit alpha-1 is a master
gene of osteoblast differentiation.
• It
upregulates osteoblast-related
genes such as
ColIA1, ALP, BSP, BGLAP, OCN
Progression of osteoblasts

• Once a pool of osteoblast progenitors

expressing Runx2 and ColIA1 has been
established during osteoblast differentiation,
there is a proliferation phase
• ALP activity
• Expression
of Osx (osterix), secretion of
OCN, BSP, collagen type I
• Become large and cuboidal
Progression towards matured
osteoblasts

Proliferation
Differentiation (preosteoblasts)

Maturation and
morphological
changes
Synthesis of bone matrix by
osteoblasts

• Two steps
• Deposition of organic matrix
• Mineralization
Deposition of bone matrix
• Osteoblasts secrets type I collagen, non
collagen proteins and proteoglycans
including decorin and biglycan which
form the organic matrix

• Mineralization then takes place in two

steps :
I. Vesicular phase

II. Fibrillar phase

Vesicular phase
Matrix vesicles are released from the apical membrane domain of
osteoblasts into newly formed bone matrix

Bind to proteoglycans and other organic components. Calcium

immobilized but later get free and get into the bone matrix

Phosphate containing compounds are degraded by ALP

and calcium and phosphate nucleate

Formation of hydroxyapatite crystals Ca10(PO4)6(OH)2 

Fibrillar Phase

• Fibrillar phase occurs when the super

saturation of calcium and phosphate
ions inside the matrix vesicles leads
to the rupture of these structures and
the hydroxyapatite crystals spread to
the surrounding matrix.
Histology of osteoblasts

• appear as a single layer of cuboidal

cells containing abundant rough
endoplasmic reticulum,
mitochondria and large Golgi
complex
• round/ovoid structures containing
dense bodies and TUNEL-positive
structures have been observed inside
osteoblast vacuoles
Histological Features continued

• Metabolically active cells are cuboidal and

less active cells are flattened.
• Osteoblastsquantity is inversely
proportional to marrow adipocytes, that
form the marrow adipose tissue.
Osteoblasts produce
• Alkaline phosphatase
• Osteocalcin (stimulated by 1,25 dihydroxyvitamin D)
• Type I collagen
• Bone sialoprotein
• Receptor activator of nuclear factor (NF)-κB ligand
(RANKL)
• Osteoprotegrin—binds RANKL to limit its activity
Receptor effector interaction in osteoblasts
Types of osteoblasts
1. Active osteoblasts: during active bone
formation, the surface layers of
osteoblasts consists of cuboidal cells,
called as active osteoblasts.

2. Inactive osteoblasts: when the bone

forming unit is not actively synthesizing
bone, surface osteoblasts are flattened,
called as inactive osteoblasts. Maintain
ionic milieu of bone.
Fate of osteoblasts
At the end of the bone‐forming phase,
osteoblasts can have one of four different fates:
1. become embedded in the bone as osteocytes
2. transform into inactive osteoblasts and
become bone‐lining cells
3. undergo apoptosis
4. transdifferentiate into cells that deposit
chondroid or chondroid bone 
Regulators of osteoblasts

activity inhibited Wnts promote

by tumor necrosis osteoblast survival
factor (TNF)-α and proliferation.

Hydrogen peroxide
Povidone-iodine (Betadine)
Bacitracin
Osteocytes

• comprise 90–95% of the total bone

cells with a lifespan of up to 25
years.
• derived
from osteoblasts
(Gegenbauer, 1864)
• locatedwithin lacunae surrounded
by mineralized bone matrix, wherein
they show a dendritic morphology
Histological features of
osteocytes
• osteocytesfrom trabecular bone are more
rounded while those from cortical bone are
elongated.
• High nucleus/cytoplasm ratio
• Longinterconnecting cytoplasmic
processes projecting through the canaliculi
From osteoblasts to osteocytes
• four
recognizable stages have been
proposed: osteoid-osteocyte, preosteocyte,
young osteocyte, and mature osteocyte.
• Process is accompanied by conspicuous
morphological and ultrastructural changes
like reduction in size, decreased RER and
Golgi Apparatus and increased N:C ratio.
Osteocyte Markers
• dentine matrix protein 1 (DMP1)
• Sclerostin
• Besides osteocytes also produce several
secondary messengers, for example, ATP,
nitric oxide (NO), Ca2+, and prostaglandins
(PGE2 and PGI2,) 
Sclerostin
 are produced by osteocytes
 help negative feedback on osteoblast
bone deposition
 is differentially down regulated based on
mechanical loading with reduced
sclerostin in areas of concentrated strain.
 down regulation of sclerostin is
associated with increased bone formation.
Function of osteocytes
strain and stress sensors, signals that are
very important for maintaining bone
structure (by virtue of lacunocanalicular
system) --> orchestrators of bone
remodeling.
Osteoclasts

• terminally differentiated multinucleated

cells, which originate from mononuclear
cells of the hematopoietic stem cell
lineage
• Factors
that influence osteoclast
formation:
• M-CSF (binds to cFMS receptor )
• RANK Ligand (binds to RANK receptor )
• Osteoprotegerin (OPG) -- > binds to RANKL
More about the RANK/ RANKL system
• RANKL/RANK interaction also promotes
the expression of other osteoclastogenic
factors such as NFATc1 and DC-STAMP
• NFATc1 interacts with transcription factors
and regulates osteoclast specific genes like
TRAP and cathepsin K
• DC-STAMP is crucial for osteoclast fusion
• Osteoclastogenesis has been demonstrated
to be dynamic
Process of bone resorption

• Osteoclasts come in contact with the extracellular

(non-collagenous)mineralized matrix(mediated by
 α v β3-integrin, CD44 )
• They then polarize and form four types of domains
i. Sealing (clear) zone [formed by actin, talin, vinculin,
paxillin, tensin and other actin associated proteins]
ii. Ruffled border
iii. Basolateral
iv. Functional
Bone resorption ..
• Ruffled border vacuolar-type H+-ATPase
(V-ATPase), which helps to acidify the
resorption (Howship) lacuna and hence to
enable dissolution of hydroxyapatite
crystals 
• After dissolution of mineral phase,
osteoclast (Oc) releases cathepsin (Cp),
matrix metalloproteinase-9 (MMP-9), and
tartrate-resistant acid phosphatase (TRAP)
that degrade the organic matrix. 
Relation between osteoblasts
and osteoclasts
• EphrinB2 (Eph2) present in osteoclast
membrane binds to ephrinB4 (Eph4) in
osteoblast (Ob) membrane, promoting its
differentiation
•  reversesignaling (ephrinB4/ephrinB2)
inhibits osteoclastogenesis
• Sema4D produced by osteoclasts inhibits
osteoblasts, while Sema3A secreted by
osteoblasts inhibits osteoclasts
Abnormal increase in
osteoclastic activity results in
• Osteoporosis
• Periarticular erosions in bony metastases
• Painful osteolytic lesion in inflammatory
arthritis
Decrease in osteoclastic activity
result in
• Osteopetrosisresulting in a
disproportionate accumulation of bone
mass
Recent advances
• It
has been shown that osteoclasts produce
factors called clastokines that control
osteoblast during the bone remodeling
cycle
• Osteoclasts
may also directly regulate the
hematopoietic stem cell niche
Signaling molecules for
osteoclasts
• IL-1 : potent stimulator of osteoclastic bone
resorption.

• IL- 10: suppresses osteoclasts

• Calcitonin: inhibits osteoclastic bone resorption

• Bisphosphonates: inhibit osteoblastic resorption

Bone Lining Cells
• quiescentflat-shaped osteoblasts that
cover the bone surfaces, where neither
bone resorption nor bone formation occurs
• exhibita thin and flat nuclear profile;
display few cytoplasmic organelle
• secretory
activity of bone lining cells
depends on the bone physiological status
• Function are not completely understood
• it
has been shown that these cells prevent
the direct interaction between osteoclasts
and bone matrix, when bone resorption
should not occur, and also participate in
osteoclast differentiation, producing
osteoprotegerin (OPG) and RANKL.
• importantcomponent of the BMU, an
anatomical structure that is present during
the bone remodeling cycle 
OSTEOPROGENITOR CELLS
• Are the undifferentiated cells or the stem cells.
• Are derived from the mesenchymal stem cells.
• They form the osteoblasts, osteocytes and the
chondrocytes.
• Become osteoblasts under low strain and
increased oxygen tension, cartilage under
intermediate strain and low oxygen tension and
fibrous tissue under high strain
• Line Haversian canals, endosteum and periosteum
• Awaiting the stimulus to differentiate
 Bone Matrix
• Is
laid down by osteoblasts which secrete both collagen and ground
substance
• Has organic and inorganic components
• Organic is 40% of dry weight of bone
• Inorganic is 60% of dry weight of bone.

Organic component:
• Major organic component is collagen (90%). Collagen gives tensile
strength to the bone and is primarily type I collagen.
Two types of bone can be identified microscopically according to the
arrangement of collagen: woven and lamellar.

i. Woven bone (also known as fibrous bone)

• characterized by a haphazard organization of collagen fibers, thus c/a woven
and is mechanically weak
• Forms quickly
• Found in all fetal bones initially, Paget's disease and in fracture healing.

ii. Lamellar bone

• has a regular parallel alignment of collagen into sheets ("lamellae") and is
mechanically strong
• Alternating layers of collagen run in opposite direction; assists bone’s ability to
resist torsion forces
remainder of the matrix is homogenous liquid
called ground substance consisting
of proteoglycans such as hyaluronic
acid and chondroitin sulfate, as well as non-
collagenous proteins such
as osteocalcin, osteopontin or bone sialoprotein
 Disorders of bone cells
• Osteoporosis • Osteopoikilosis (Spotted Bone
disease)
• Rickets & Osteomalacia
• Osteitis Deformans
• Osteogenesis imperfecta
• Osteochondrosis
• Osteopetrosis
• Osteonecrosis
• Lead poisoning
• Scurvy
• Secondary hyperparathyroidism
• Malignancy – myeloma,
leukemia
Osteoporosis
• A condition characterized by age related
decrease in the bone mass . It is a
quantitative defect.
References
1. Florencio-Silva R, Sasso GR, Sasso-Cerri E,
Simões MJ, Cerri PS. Biology of Bone
Tissue: Structure, Function, and Factors That
Influence Bone Cells. Biomed Res Int.
2015;2015:421746.
doi:10.1155/2015/421746
2. Miller’s Review of Orthopaedics , Seventh
Edition, Elsevier, 2016
3. Wikipedia
4. Google Images
THAN
K YOU