CELL INJURY AND DEATH

By:
Taleya HidayatUllah
 Cell injury
• Results when cells are stressed so severely
that they are no longer able to adapt or when
cells are exposed to damaging agents.
• Cell injury can be reversible or irreversible.
 Reversible cell injury
• Functional and morphologic changes are
reversible if the damaging stimulus is
removed.
• The features are: decreased oxidative
phosphorylation, ATP depletion and cellular
swelling.
 Irreversible injury and cell death.
• With continuing damage,injury becomes
irreversible.
• Cells undergo morphologic changes
recognisable as cell death.
• Cell death is of 2 types-necrosis and apoptosis.
Reversible and irreversible injury
 Causes of cell injury
• Oxygen deprivation(hypoxia)
a. Ischemia
b. anemia
• Physical agents eg:mechanical trauma,burns,deep cold,electric
shock.
• Chemical agents & drugs eg:poisons,environmental
pollutants,CO,asbestos,alcohol,narcotic drugs etc.
• Infectious agents-viruses,rickettsiae,bacteria,fungi,protozoa
and helminths.
• Immunologic reactions-anaphylaxis,autoimmune disorders.
• Genetic derangements.
• Nutritional imbalances: obesity,specific vitamin deficiencies
etc.
 Mechanisms of cell injury
Cell systems are particularly vulnerable to injury
• Depletion of ATP-affects activity of Na,K-ATPase
pump.
• Mitochondrial damage-leakage of cytochrome-C into
cytosol,resulting in apoptosis.
• Influx of Ca & loss of Ca homeostasis,leading to
activation of ATPases,phospholipases,proteases &
endonucleases.
• Depletion of ATP-affects activity of Na,K-ATPase pump.This
results in anaerobic glycolysis.
• Mitochondrial damage
• loss of Ca homeostasis
 Cellular enzymes involved in cell injury
ATPASES
• Hasten ATP depletion.
Phospholipases
• cause membrane damage.
Proteases
• breakdown membrane & cytoskeletal
proteins.
Endonucleases
• cause DNA & protein fragmentation.
INJURY DUE TO FREE RADICALS
 Free radical injury
• Free radical – atoms or groups of atoms with
an odd (unpaired) number of electrons
• Energy created by this unstable configuration
is released through reactions with adjacent
molecules.
• They initiate autocatalytic reactions.
 Free radicals are initiated or
generated by:
• Absorption of radiant energy(u-v or ionising):Water
is hydrolysed to(OH)&(H) free radicals.
• Enzymatic metabolism of exogenous chemicals or
drugs ,eg:CCl4 converted to CCl3.
• Redox reactions in the cell: (O2),(H2O2) &(OH).
• Transition metals like iron and copper.
• Nitrous oxide.
 Effects of free radicals:
• Lipid peroxidation of membranes:
Polyunsaturated fatty acid of membrane is attacked
repeatedly by free radicals to form highly destructive
polyunsaturated fatty acid (PUFA) radicals.

This is termed as lipid peroxidation. These lipids are widely

spreaded to other part of membrane that is lipid peroxidation
takes place at adjoining part of membrane causing damage to
entire cell membrane.
• Oxidation of protein: Free radical causes CELL
INJURY by oxidation of protein
macromolecules of cell causing cross linkage
in the amino acid sequences of protein and
fragmentation of polypeptides
• Effect on DNA damage: Free radical breaks
DNA fragments to single strand, so there will
be formation of DNA which is defective.
Replication of this DNA is not possible and
there by cell death may occur.
• Cytoskeleton Damage: Free radicals interfere
with mitochondrial aerobic phosphorylation
and decreases synthesis of ATP leading to
cytoskeleton damage.
 Inactivation of free radical
reactions.
• Can be enzymatic or non-enzymatic.
• Enzymatic: catalase,superoxide
dismutases,glutathione peroxide.
• Non-enzymatic:-Antioxidants eg: Vitamins A,E,C
& glutathione .
• Iron,copper can catalyze the formation of reactive
oxygen species. The levels of these reactive forms
are minimized by binding of the ions to storage
and transport proteins (Transferrin, Ferritin,
Lactoferrin and ceruloplasmin) thereby
minimizing OH formation.. .
 Earliest changes in reversible cell
injury are:
• Decreased generation of ATP.
• Loss of cell membrane integrity.
• Defects of protein synthesis.
• Cytoskeletal damage.
• DNA damage.

• Within limits,the cell can compensate for these

derangements.Persistent or excessive injury leads to
irreversible injury.
 Ischemia

Cell oxygen tension Membrane injury

Loss of phospholipids,
Increase of free radicals
Reduced ATP Lipid breakdown

Incr. Glycolysis
Decr. Protein systhesis Leakage of cell enzymes
Decr glycogen
Lipid deposition Calcium influx

Intracellular lysosomal
enzyme release
Reversible injury:
cell swelling
microvlli loss Irreversible injury
blebs Reduced basophilia
ER swelling
Nuclear changes
myelin figures
chromatin clumping Protein digestion
 Characteristic phenomena of
irreversibility:
• Inability to reverse mitochondrial dysfunction.
• Development of profound disturbances in membrane
function.
• Therefore,in cardiac muscle death there is leakage of
CKMB & troponin.
• In injury to bile duct epithelium & liver,serum
alkaline phosphatase is raised.
• In hepatocyte injury,transaminases are raised.
 Necrosis
• Necrosis can be defined as cell death caused
by loss of membrane integrity, intracellular
organelle swelling and adenosine triphosphate
(ATP) depletion leading to an influx of calcium.
 Necrosis
• A spectrum of morphologic changes that follow
cell death in living tissue,due to progressive
degradative action of enzymes on the lethally
injured cells.
• Leaked out contents of necrotic cells may elicit
inflammation in the surrounding tissue.
• The morphologic appearance is due to
denaturation of proteins and enzymatic digestion.
• The enzymes are derived from lysosomes of the
dead cells themselves-AUTOLYSIS.
 Pathology of necrotic cells
• Increased eosinophilia with a glassy homogeneous
appearance as a result of glycogen particles.
• Moth-eaten appearance in cells with vacuolated
cytoplasm due to enzymatic digestion of cytoplasmic
organelles.
• Calcification of dead cells.
• Replacement of dead cells by whorled phospholipid
masses called myelin figures(ultrastructurally).
Necrosis
Trauma (toxic chemicals, mechanical injury, heat, hypoxia)
Loss of ability to regulate internal environment
Ca2+ influx accompanied by swelling
Alteration of protein activity
calpain
cathepsin
caspase
Production of toxic compounds
(activation of cyclooxygenases)
arachadonic acid
prostaglandins
eicosanoids

Inflammation
 Nuclear changes in necrotic cells:
• Karyolysis-basophilia of chromatin may fade.
• Pyknosis-nuclear shrinkage and increased
basophilia.
• Karyorrhexis-nuclear fragmentation.
• Disappearance of nucleus.
Patterns
– Coagulative
– Liquefactive
– Caseous
– Fat necrosis
– (gangrene)
– (Infarct)
• Red/haemorrhagic
• White
 Coagulative necrosis
• Denaturation of protein is the primary
pattern.
• Basic cell outline is preserved for some days
eg:acute MI.

Normal Necrosis
 Liquefactive necrosis
• Complete dissolution of necrotic tissue.
• Most commonly due to massive infiltration by
neutrophils (abscess formation).
– Release of reactive oxygen species and proteases
• Liquefaction is also characteristic of ischaemic
necrosis in the brain.
 Caseous necrosis
• Accumulation of amorphous (no structure)
debris within an area of necrosis.
• Tissue architecture is abolished and viable
cells are no longer recognizable.
• Characteristically associated with the
granulomatous inflammation of tuberculosis.
Also seen in some fungal infections.
 Fat necrosis
• Results from the action of lipases released into
adipose tissue.
– pancreatitis, trauma.
• Free fatty acids accumulate and precipitate as
calcium soaps (saponification).
– These precipitates are grossly visible as pale yellow/white
nodules
• Microscopically, the digested fat loses its cellular
outlines. There is often local inflammation
 Gangrene ("gangrenous necrosis")
• Not a separate kind of necrosis at all, but a term for
necrosis that is advanced and visible grossly.
– If there's mostly coagulation necrosis, (i.e., the typical
blackening, desiccating foot which dried up before the
bacteria could overgrow), we call it dry gangrene.
– If there's mostly liquefactive necrosis (i.e., the typical foul-
smelling, oozing foot infected with several different kinds
of bacteria), or if it's in a wet body cavity, we call it wet
gangrene.
 Infarction
• An area of ischaemic necrosis in a tissue or
organ
– White
• Arterial occlusion in most solid tissues
– Red/haemorrhagic
• Venous occlusion
• Loose tissues
• Dual blood supply
• Previously congested
 Apoptosis
• Programmed cell death.
• Noxious stimuli that damage DNA result in
nuclear dissolution without complete loss of
cell membrane integrity.
• Can be physiologic or pathologic.
 Apoptosis - triggers
• Withdrawal of growth stimuli
– E.g. growth factors
• Death signals
– E.g. TNF and Fas
• DNA damage
– p53 plays an important role
 Apoptosis - mechanisms
• Extrinsic factors
– E.g. by members of the TNF family
• Intrinsic mechanisms
– E.g. hormone withdrawal
What makes a cell commit suicide?
• withdrawal of positive signals (growth factors, Il-2)
• receipt of negative signals (increased levels of oxidants, DNA damage via X-ray or UV light,
chemotherapeutic drugs, accumulation of improperly folded proteins, death activators such
as: TNF-a, TNF-b, Fas/FasL)

Steps in apoptosis:
the decision to activate the pathway;
the actual "suicide" of the cell;
engulfment of the cell remains by specialized immune cells called phagocytes;
degradation of engulfed cell.
The actual steps in cell death require:
condensing of the cell nucleus and breaking it into pieces
condensing and fragmenting of cytoplasm into membrane bound apoptotic bodies;
breaking chromosomes into fragments containing multiple number of nucleosomes (a
nucleosome ladder)
Apoptosis Triggered via Two Pathways
Intrinsic or mitochondrial pathway
Extrinsic or death receptor pathway