Cellular pathology

Cell Injury ,Cellular Adaptations, Cell Death and

intracellular accumulations

Zemen A.,MD,Pathologist

DTU college of medicine and health sciences

March 2021
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BASIC CONCEPTS IN CYTOLOGY

Introduction

Components of the cell

Cell membrane

Cytoplasm

Organelles

Cell division
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 Introduction
Multicellular organisms are made up of
cells
intercellular substances
body fluids
Cells are the fundamental units of structure and
function of an organism
100 trillion cells in a human
200 kinds of cells
all have certain structural features in common
Cells perform all activities necessary for survival, growth
and reproduction of an organisms
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Components of cells
Cells are anatomically partitioned into 2 major
compartments
Nucleus - composed of nucleoplasm (karyoplasm)
Cytoplasm - comprises

Cytosol (cytoplasmic matrix)

Cytoskeleton - fibrillar components

Organelles

Inclusions
Cells are enclosed in plasma (cell) membrane

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 Cell membrane

• Flexible outer layer; about 7.5 nm thick

• Composed of a lipid bilayer and associated proteins
• Functions
• semi permeable membrane
• a gatekeeper that regulate passage of
substances into and out of the cell
• sensory devise that permits the cell to recognize & be
recognized by other cells & macromolecules
• cell-to-cell attachment & communication

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Fluid mosaic model of membrane structure

Describes the molecular arrangement of the plasma

membrane

According to this model, the membrane is a mosaic

of proteins floating like icebergs in a sea of lipids

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 Membrane lipids
Responsible for permeability properties of membrane

Composed of

phospholipids
glycolipids
cholesterol

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Membrane lipids: Phospholipids (75%)
• Line up in two parallel rows forming phospholipid bilayer
• This arrangement occur because
– The phospholipids are amphipathic i.e. they posses polar &
non-polar regions
• Polar
– phosphate containing ‘head’
– hydrophilic (mixes with water)
– faces the membrane surface
• Non-polar
– two fatty acid ‘tails’
– hydrophobic (do not mix with water)
– projects into the interior of the membrane

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Plasma membrane organization and asymmetry of membrane
lipids
The phospholipid distribution within the membrane is asymmetric
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 Membrane lipids: Glycolipids (5%)
• Lipids with one or more sugar groups attached
• Amphipathic: polar carbohydrate residue extend to ECM
(glycocalyx)
• Present in the outer leaflet only
• Functions
• adhesion among cells & tissues
• cell to cell recognition & communication
• regulation of cellular growth & development

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Membrane lipids: Cholesterol (20%)

• Located in both leaflets of the plasma membrane

• The stiff steroid rings of cholesterol strengthen the
membrane but decrease its flexibility
• assist in maintaining the structural integrity of
plasma membrane

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 Fluidity of the lipid bilayer

• The phospholipid bilayer is dynamic

• the lipids can move side ways and exchange places in their
own layer
• Flip-flop of molecules between layers rarely occur
• It is also self-healing
• Important for cellular activities; exocytosis, endocytosis,
biogenesis & trafficking
• Increase by rise in temperature and greater unsaturation
of fatty acid tails
• Decrease by increase in cholesterol content

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Membrane proteins

2 types

Integral proteins
Peripheral proteins

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 Membrane proteins: Integral proteins

• Extend across the phospholipid bilayer among the fatty

acid tails
• Amphipathic
• Most are glycoproteins
• Function
Channels (pores) through which certain substances flow into
or out of the cell
Transporters (carriers) to move substance from one side of
membrane to other
Recognition sites (receptors)

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 Membrane proteins: Peripheral proteins
• Do not extend across the phospholipid bilayer
• Located on cytoplasmic aspect of inner leaflet (commonly)
• Functions
Enzymes
Cytoskeleton anchors
Intracellular messenger system
• Some membrane proteins can diffuse laterally in the lipid
bilayer, whereas others are immobile
• Ratio of lipid to protein is 1:1 (by weight) in most cells and
4:1 in myelin

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Membrane transport

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Movement of small molecules and larger structures across membranes. The lipid bilayer
is relatively impermeable to all but the smallest and/or most hydrophobic molecules.
Thus the import or export of charged species requires specific transmembrane transporter
proteins, vesicular traffic, or membrane deformations
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Cytoplasm
Contain 4 main structural components

organelles
inclusions
cytosol (fluid component)
cytoskeleton

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 Organelles
• Specialized structures that have characteristic shapes and
specific functions
• Metabolically active units of living matter
• 2 types
Membranous organelles
• cell membrane, mitochondria, RER, Golgi apparatus, secretory
vesicles, lysosomes, peroxisomes, SER
Non - membranous organelles
• ribosomes, centrioles, cilia & flagella

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Basic subcellular constituents of cells
The figure shows geographic relationships but is not intended to be
accurate
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The table presents the number of the various organelles within a
typical hepatocyte, as well as their volume within the cell.
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 Ribosomes
• Contain ribosomal RNA (rRNA) and several ribosomal proteins
• Function
– protein synthesis
• Found in 2 forms
– Free ribosomes
• float in the cytosol; have no attachments to other
organelles
• synthesize proteins for use inside the cell
– Attached ribosomes
• to endoplasmic reticulum or outer nuclear membrane
• involved in the synthesis of proteins destined for
insertion in the plasma membrane or for export from
the cell
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 Endoplasmic Reticulum
• A system of sacs or tubules (channels) bounded by
membranes of varying shape called cisterns
• Continuous with outer nuclear membrane
• Based on its association with ribosomes, the ER is divided
into 2 types
• Rough (granular) endoplasmic reticulum (RER)
• studded with ribosomes
• the site where non-cytosolic proteins are synthesized
• Smooth (agranular) endoplasmic reticulum (SER)
• with no ribosomes on its surface
• appear as branching anastomosing tubule or vesicles

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 Endoplasmic Reticulum
• Functions
• General
• provides surface area for chemical reaction
• synthesizes glycoprotiens and phospholipids
• transportation of cellular products from one
portion of cell to another
• SER
• steroid hormone synthesis - Leydig cells of testis
• drug detoxification - hepatocytes
• muscle contraction & relaxation (sarcoplasmic
reticulum)

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 Golgi apparatus
• Consists of several disk-shaped cisternae (saccules)
arranged in a stack
• Associated with cisterns are small Golgi vesicles
• Includes the following regions
– Cis (entry) face - outer convex cisternae, face RER
– Trans (exit) face - inner concave cisternae
– medial compartment - few cisterns between the two
• Functions
– processing of non cytosolic proteins synthesized in the RER
– Forms secretory vesicles
– membrane retrieval, recycling & redistribution

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 Lysosomes
• Dense, membrane - bounded vesicles
• Formed when sequestered materials fuses with a late
endosome
• Contains more than 40 powerful enzymes (hydrolases)
which are synthesized in RER
• these enzymes breakdown
- damaged cellular structures
- food particles ingested by the cell
- unwanted things like bacteria

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Intracellular catabolism. (A) Lysosomal degradation. In heterophagy (right side of panel A), lysosomes fuse with
endosomes or phagosomes to facilitate the degradation of their internalized contents . The end products may be
released into the cytosol for nutrition or discharged into the extracellular space (exocytosis). In autophagy (left
side of panel A), senescent organelles or denatured proteins are targeted for lysosome-driven degradation as they
are encircled within a double membrane vacuole derived from the endoplasmic reticulum and marked by LC3
protein (microtubule associated protein 1A/1B-light chain 3). Cell stress, such as nutrient depletion or some
intracellular
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(B) Proteosomal degradation. Cytosolic proteins destined for turnover (e.g., transcription factors or regulatory
proteins), senescent proteins, or proteins that have become denatured due to extrinsic mechanical or chemical
stresses can betagged by multiple ubiquitin molecules (through the activity of E1, E2,and E3 ubiquitin ligases). This
marks the proteins for degradation by proteasomes, cytosolic multi-subunit complexes that degrade proteins to small
peptide fragments. High levels of misfolded proteins within the endoplasmic reticulum (ER) trigger a protective
unfolded protein response—engendering a broad reduction in protein synthesis, but specific increases in chaperone
proteins that can facilitate protein refolding. If this is inadequate to cope with the levels of misfolded proteins56it can
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lead to apoptosis
 Mitochondria

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Mitochondria
Morphology
• Rod shaped
• Consists of 2 membranes
–Outer membrane - bounds the organelle,
smooth
–Inner membrane - invaginate (folds) to form
cristae
• 2 compartments
–Intermembrane compartment (between the 2
membranes)
–Inner matrix compartment (enclosed by inner
membrane and cristae)
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Function
• Generate ATP (power house of a cell)
–Site of aerobic cellular respiration
• Heat production in brown fat cell
Unique properties
possess their own genetic apparatus
proliferate by division (fission) of pre existing mitochondria
Origin
• Might have originated as symbionts (intracellular parasites)
–anaerobic eukaryotic cells endocytosed aerobic
microorganisms that evolved in to mitochondria

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 Inclusions
• ‘lifeless’ accumulations of materials that are not
metabolically active
• usually present in cytosol only temporarily
• not bounded by membrane (usually)
• Examples
• Glycogen - stored energy source
• Lipid droplets - storage form of triglycerides
and cholesterol
• Lipofusin - granular material + lipid droplets

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 Cytoskeleton
• structural frame work with in the cytosol
• network of filaments
• functions
• maintain cell shape
• stabilizing cell attachments
• promote cell motility

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 Cytoskeleton
• 3 protein filaments constitute cytoskeleton
microtubules
• composed of α and ß tubulin proteins
• function - maintain cell shape, transport, promote movement of
cilia, flagella and chromosomes
microfilaments
• composed of actin and myosin
• function - locomotion, endocytosis and exocytosis
intermediate filaments
• include keratin, desmin, vimentin, laminin
• function - provide mechanical strength

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Cytoskeletal elements and cell-cell interactions. Interepithelial adhesion involves several different surface protein
interactions at tight junctions, adherens junctions, and desmosomes; adhesion to the extracellular matrix involves
cellular integrins (and associated proteins) within hemidesmosomes. The various adhesion proteins within the plasma
membrane associate with actin microfilaments and intermediate filaments to provide a mechanical matrix for cell
structure and signaling. Gap junctions do not impart structural integrity but allow cell-cell communication by the
movement of small molecular weight and/or charged species.
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Nucleus
spherical or oval in shape (varies in shape)
varies in number
largest structure
absent in mature RBCs and platelets
double membrane (nuclear envelope) separate
it from the cytoplasm
within it there are nucleolus, nucleoplasm and
chromatin
Function
contains genetic apparatus which control cellular
structure and direct cellular activities
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Nuclear envelope
• surrounds nuclear material
• consists of 2 parallel membrane separated from each
other by a narrow perinuclear cisterna
• perforated at intervals by openings called nuclear pores
– allows most ions and water soluble molecules
to shuttle between nucleus and cytoplasm
Nucleolus
• well-defined, not membrane bounded spherical bodies
• one or more
• contain protein, DNA & RNA
• detectable in interphase
• involved in synthesis of rRNA

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Nucleoplasm
• the portion of protoplasm surrounded by nuclear envelope
• consists of matrix and various particles
Chromatin
• double - stranded DNA complexed with histones and acidic
proteins
• responsible for RNA synthesis
• 2 forms
• hetrochromatin - dense appearance;
trascriptionally inactive
• euchromatin – light; transcriptionally active

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 DNA

Chromosomes can only be visualized by light microscopy during

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The organization of nuclear DNA. At the light microscopic level, the nuclear genetic material is
organized into dispersed, transcriptionally active euchromatin or densely packed,
transcriptionally inactive heterochromatin; chromatin can also be mechanically connected
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nuclear membrane, and nuclear membrane perturbation can thus influence 116
transcription
Noncoding DNA
The human genome contains roughly 3.2 billion
DNA base pairs.
Within the genome there are about 20,000 protein-
encoding genes, comprising only about 1.5% of the
genome.
These proteins variously function as enzymes,
structural components, and signaling molecules and
are used to assemble and maintain all of the cells in
the body

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98.5% of the human genome that does not encode
proteins.
 It has been known for some time that protein-
coding genes in higher organisms are separated by
long stretches of DNA whose function has been
obscure for many years—sometime denoted as
“dark matter” of the genome

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NONCODING REGIONS OF DNA
Promoters are noncoding regions of DNA that initiate gene
transcription
Enhancers are regulatory elements that can modulate gene
expression over distances of 100 kB or more by looping back
onto promoters and recruiting additional factors that are needed
to drive the expression of pre-mRNA species.
The intronic sequences are subsequently spliced out of the pre-
mRNA to produce the definitive message that is translated into
protein—without the 3′- and 5′-untranslated regions (UTR).
In addition to the enhancer, promoter, and UTR sequences,
noncoding elements are found throughout the genome; these
include short repeats, regulatory factor binding regions,
noncoding regulatory RNAs, and transposons

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The major classes of functional non–protein-coding
sequences found in the human genome are the
following
1. Promoter and enhancer regions that provide binding
sites for transcription factors
2. Binding sites for factors that organize and maintain
higher order chromatin structures
3. Noncoding regulatory RNAs. More than 60% of the
genome is transcribed into RNAs that are never translated
into protein, but which nevertheless can regulate gene
expression through a variety of mechanisms. The two best-
studied varieties—micro-RNAs and long noncoding RNAs

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Mobile genetic elements (e.g., transposons).
Remarkably, more than one third of the human
genome is composed of these elements, popularly
denoted as “jumping genes.” These segments can
move around the genome, exhibiting wide variation
in number and positioning even amongst closely
related species (i.e., humans and other primates).
They are implicated in gene regulation and
chromatin organization, but their function is still not
well established.
Special structural regions of DNA, in particular
telomeres (chromosome ends) and centromeres
(chromosome “tethers”)

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The telomeres are repetitive nucleotide sequences
that cap the termini of chromatids and permit
repeated chromosomal replication without loss of
DNA at the chromosome ends

The centromeres act as the locus for the formation

of a kinetochore protein complex that regulates
chromosome segregation at metaphase

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Noncoding DNA in health and diseases
Many, and perhaps most, of the genetic variations
(polymorphisms) associated with diseases are located in
non–protein-coding regions of the genome.

Thus, variation in gene regulation may prove to be more

important in disease causation than structural changes in
specific proteins.

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Histone organization. A, Nucleosomes are comprised of octamers of histone proteins (two
each of histone subunits H2A, H2B, H3, and H4) encircled by 1.8 loops of 147 base pairs
of DNA; histone H1 sits on the 20-80 nucleotide linker DNA between nucleosomes and
helps stabilizes the overall chromatin architecture. The histone subunits are positively
charged, thus allowing the compaction of the negatively charged DNA
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B, The relative state of DNA unwinding (and thus access for transcription factors) is
regulated by histone modification, for example, by acetylation, methylation, and/or
phosphorylation (so-called “marks”); marks are dynamically written and erased. Certain
marks such as histone acetylation “open up” the chromatin structure, whereas others, such
as methylation of particular histone residues, tends to condense the DNA and leads to gene
silencing. DNA itself can also be also be methylated, a modification
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Chromatin patterns

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Genome-Wide Identification of Chromatin Trans

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 General Concepts of Genetics
• Genetic information for protein synthesis is stored in
DNA molecules contained in the nucleus
• All cells have the same genetic information but each
uses only a portion of this information depending on its
structure and function

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 DNA
• DNA – made up of
nucleotides which
consists of Phosphoric
acid, five carbon sugar
(deoxyribose) and
one of the four
nitrogenous bases
(A,G,T,C)

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Protein Synthesis

RNA is involved in protein synthesis

Messenger RNA-transcribes instructions from the
DNA to Cytoplasm
Ribosomal RNA- machinery needed for protein
synthesis (translation)
Transfer RNA- delivery of appropriate Amino acid to
ribosome
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Gene silencing

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Noncoding RNAs
As the name implies, these are encoded by genes
that are transcribed but not translated.

Although many distinct families of noncoding

RNAs exist, we will only discuss two examples
here: small RNA molecules called microRNAs, and
long noncoding RNAs >200 nucleotides in length.

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MicroRNA and Long Noncoding RNA

• miRNA are short RNAs (21 to 30 nucleotides); they do not

encode proteins but rather are involved in posttranscriptional
silencing of gene expression.
• miRNA transcription produces a primary miRNA, which is
progressively trimmed by the enzyme DICER, eventually
becoming associated with a multiprotein aggregate called
RNA-induced silencing complex (RISC).
• Subsequent base pairing between the miRNA strand and its
target mRNA directs the RISC to either induce mRNA
cleavage or repress its translation

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• Synthetic small interfering RNAs (siRNAs) are short RNA
sequences that can be introduced into cells, acting in a
manner analogous to endogenous miRNAs.

• These form the basis for knockdown experiments to study

gene function and are also being developed as possible
therapeutic agents to silence pathogenic genes

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Long noncoding RNA (lncRNA)
• Long noncoding RNA (lncRNA) exceed coding mRNAs by
tenfold to twentyfold and are involved in modulating gene
expression:
• • lncRNA can restrict RNA polymerase access to specific
coding genes.
The best example involves XIST, which is transcribed from the X
chromosome and plays an essential role in physiologic X
chromosome inactivation.
• lncRNA can facilitate transcription factor binding, thus
promoting gene activation.
• lncRNA can facilitate chromatin modification or provide the
scaffolding to stabilize chromatin structure

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Protein Synthesis

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• Guanine, Adenine, Cytosine and thyamine are “the
alphabets of the genetic code”
• Triplets of these is called a codon
• Example – GUC codes for Amino acid Alanine
• There are three stop codones

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• A gene – Several hundred to almost a million base
pairs can represent a gene

• Humans have around 30,000 genes

• DNA molecule is combined with several types of

proteins and small amount of RNA and is visible
under the microscope as chromatin.
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QUESTION
• Humans have 99.5% DNA Sequence resemblance
• What brought so huge phenotypic difference among humans
• Is it the 0.5% difference in DNA sequence or something
else?
• Genetic-0.5% accounts about 15 million base pairs
 DNA Variations in this segments of DNA are important.
 DNA variation-SNP AND CNVs
• Epigenetics
• Histone modifications
 Chromatin writers and erasers
 Histone methylation, acetylation and phosphorylation
 DNA methylation

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Cellular Adaptations
Cell Injury
Cell Death

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OVERVIEW OF CELLULAR RESPONSES TO
STRESS AND NOXIOUS STIMULI
Homeostasis
a state of equilibrium within a cell, tissue, organ, organ
system or organism.
The normal cell is confined to a fairly narrow range
of function and structure
-By its genetic programs of metabolism, differentiation,
and specialization;
– By constraints of neighboring cells; and
– By the availability of metabolic substrates
Normal cells handle normal physiologic demands
and maintain a steady state
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 If the limits of adaptive
responses are exceeded

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Adaptation
Occurs when cells encounter more severe
physiologic stresses and some pathologic stimuli
 These stresses bring about a number of
physiologic and morphologic cellular adaptations
Adapted cells achieve a new but altered steady
state
– preserving the viability of the cell
– modulating its function as it responds to such
stimuli

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Cell Injury

Definition- Molecular or structural alteration in a cell

that may or may not lead to cell death

• Occur after a sequence of events by the time

adaptive responses are limited or exceeded or
when adaptation is not possible.

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Cell Death
A final turning point of cell injury

One of the most crucial events in the evolution of

disease of any tissue or organ

Affects every cell type

There are 2 patterns of cell death

–Apoptosis
–Necrosis
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 CELLULAR ADAPTATIONS
 Alterations in cell size

 Alterations in cell number

 Alterations in cell differentiation

 Abnormal intracellular accumulations

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 Cellular adaptation
• Cellular adaptations
• Brought about by more severe physiologic stresses and some
pathologic stimuli
• The adaptive response may consist of
• Hyperplasia - an increase in the number of cells
• Hypertrophy -an increase in the sizes of individual cells
• Atrophy - a decrease in the size and function of cells
• Metaplasia - a change of cells from one type to another
• Cell injury is reversible up to a certain point
• If the stimulus persists or is severe enough from the beginning, the
cell reaches a “point of no return” and suffers irreversible cell
injury and ultimately cell death.

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 HYPERPLASIA;

• Hyperplasia is an increase in the number of cells in an organ or tissue,

usually resulting in increased volume of the organ or tissue.
• Hyperplasia can be physiologic or pathologic.

04/06/2021 262
• Hyperplasia is generally caused by increased local production of
growth factors, increased levels of growth factor receptors on the
responding cells, or activation of particular intracellular signaling
pathways.
• Pathologic Hyperplasia
• Most forms of pathologic hyperplasia are caused by excessive
hormonal stimulation or growth factors acting on target cells.
• Endometrial hyperplasia and benign prostatic hyperplasia are an
example of abnormal hormone-induced hyperplasia
• Pathologic hyperplasia constitutes a fertile soil in which cancerous
proliferation may eventually arise.

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 C

• Uterus in normal A. Gravid uterus gross,; B,Histology in normal condition and C, Histology in
pregnancy

04/06/2021 266
 Mechanisms of Hyperplasia

Stress or insult
Local production of growth factors.

Increased level of growth

factor receptors in cells.

Activation of intracellular signaling

04/06/2021
pathways. 267
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• Mechanism of hypertrophy involve many signal
transduction pathways, leading to the induction of a
number of genes, which in turn stimulate synthesis of
numerous cellular proteins.
• -Physiologic
• E.g –Uterus,skeletal muscle
• Pathologic -heart muscle
•

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 Left Ventricle

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 ATROPHY

Physiologic atrophy is common during early

development.

Pathologic atrophy depends on the underlying

cause and can be local or generalized

04/06/2021 273
 Causes of atrophy are;
- Decreased workload (atrophy of disuse).
- Loss of innervation (denervation atrophy).
-Diminished blood supply
-Inadequate nutrition
-Loss of endocrine stimulation
-Aging (senile atrophy).
-Pressure.

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Muscle ischemic atrophy:

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 METAPLASIA

• It may represent an adaptive substitution of cells that are

sensitive to stress by cell types better able to withstand the
adverse environment.

• The most common epithelial metaplasia is columnar to

squamous, as occurs in the respiratory tract in response to
chronic irritation

04/06/2021 282
• the influences that predispose to metaplasia, if persistent,
may induce malignant transformation in metaplastic
epithelium
• Metaplasia from squamous to columnar type may also
occur, as in Barrett esophagus following reflux esophagitis,
may result in adenocarcinoma
• Metaplasia occurs by the differentiation of stem cells to a
particular lineage signals generated by cytokines,growth
factors, and extracellular matrix components in the cell’s
environment

04/06/2021 283
Metaplasia of normal columnar (left) to squamous epithelium (right) in a bronchus,
shown (A) schematically and (B) histologically
04/06/2021 284
Disadvantage of metaplasia
In all these instances, the more rugged stratified squamous
epithelium is able to survive under circumstances in which
the more fragile specialized columnar epithelium most
likely would have succumbed.
Although the metaplastic squamous cells in the respiratory
tract, for example, are capable of surviving, an important
protective mechanism—mucus secretion—is lost.
Moreover, the influences that predispose to metaplasia, if
persistent, may induce malignant transformation in
metaplastic epithelium.
Thus, the common form of cancer in the respiratory tract is
composed of squamous cells, which arise in areas of
metaplasia of the normal columnar epithelium into
squamous epithelium
04/06/2021 285
Cont’d…
• Metaplasia from squamous to columnar type may also
occur (glandular metaplasia), eg. Barrett esophagus
• In which the esophageal squamous epithelium is replaced by
intestinal-like columnar cells under the influence of refluxed
gastric acid .
• Cancers may arise in these areas, and these are typically
glandular (adeno) carcinomas

• Connective tissue metaplasia: eg. the formation of

cartilage, bone, or adipose tissue (mesenchymal tissues)
in tissues that normally do not contain these elements.
• Myositis ossificans: bone formation in skeletal muscle
• Osteoma cutis: bone formation in subcutis

04/06/2021 286
• Metaplasia. B, Metaplastic transformation of esophageal
stratified squamous epithelium (left) to mature columnar
epithelium (so-called Barrett metaplasia).
04/06/2021 287
Mechanisms of Metaplasia
• Metaplasia does not result from a change in the
phenotype of the differentiated cell types
• Instead it is the result of a reprogramming of stem
cells that are known to exist in normal tissues, or of
undifferentiated mesenchymal cells present in
connective tissue.
• The differentiation of stem cells to a particular lineage is
brought about by signals generated by cytokines, growth
factors, and extra cellular matrix components in the
cell's environment.

04/06/2021 288
Dysplasia
• Deranged cell growth of specific tissue that result in
to cells that vary in size ,shape ,appreance.
• Even though dysplasia is abnormal ,but potentially
reversible if the cause removed.
• Dyspastic tissue change may progress to neoplastic
disease.

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 CELL INJURY

04/06/2021 295
CELL INJURY AND CELL DEATH
cell injury results when cells are stressed so severely that they are no
longer able to adapt or when cells are exposed to inherently
damaging agents.

Reversible cell injury- is manifested as functional and morphologic

changes that are reversible if the damaging stimulus is removed.

The hallmarks of reversible injury are; reduced oxidative

phosphorylation, adenosine triphosphate (ATP) depletion, and
cellular swelling caused by changes in ion concentrations and water
influx.

04/06/2021 296
Reversible injury
• Two patterns of reversible cell injury
cellular swelling
fatty change
• Cellular swelling
–failure of ionic and fluid homeostasis due to loss of
function of energy dependent ion pumps
• Fatty change
-in hypoxic and toxic or metabolic injury
-manifest by the appearance of small or large lipid
vacuoles in the cytoplasm
-principally occur in cells involved in and dependent on
fat metabolism such as hepatocytes, myocardial cells
04/06/2021 297
Other reversible changes of cell
injury
• Stresses of different types may induce changes in cells and
tissues other than adaptations and death.
• Cells that are exposed to sublethal or chronic stimuli
may not be damaged but may show a variety of
subcellular alterations.

• Metabolic derangements in cells may be associated with

intracellular accumulations of a number of substances:
pigments, proteins, lipids, and carbohydrates.

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• Cellular swelling may be difficult to appreciate light
microscopically
• When it affects many cells in an organ it causes some
pallor, increased turgor and weight
• Light microscope – small clear vacuoles may be seen
Ultrastructural changes
1. plasma membrane alterations, such as
blebbing,blunting, and distortion of microvilli;

04/06/2021 300
2. mitochondrial changes, including swelling,
the appearance of small densities
3. dilation of the endoplasmic reticulum, with
detachment and disaggregation of polysomes
4. nuclear alterations, with disaggregation of
granular and fibrillar elements

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• Irreversible injury and cell death - occurs with continuing
damage to the extent that the cell cannot recover at all
• Irreversibly injured cells invariably undergo morphologic
changes that are recognized as cell death.
• There are two types of cell death, necrosis and apoptosis
• Necrosis is characterized by severe damage to membrane,
entry of lysosomal enzymes in to the cytoplasm and
digesting the cell and leakage of cellular content in to the
extracellular space while,
• Apoptosis is characterized by nuclear dissolution without
complete loss of membrane integrity.
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04/06/2021 303
1.Oxygen Deprivation/ Hypoxia /
• Hypoxia is the most common cause of injury; it
occurs when lack of oxygen prevents the cell from
synthesizing sufficient ATP by aerobic oxidation.
• Major mechanisms leading to hypoxia are ischemia,
cardiopulmonary failure, and decreased oxygen-
carrying capacity of the blood (e.g., anemia).
• Ischemia, due to a loss of blood supply, is the most
common cause of hypoxia, and is typically related to
decreased arterial flow or decreased venous outflow
(e.g., atherosclerosis, thrombus, thromboembolus)

04/06/2021 304
- 2.Physical Agents- include mechanical trauma,
extremes of temperature(burns and deep cold),
sudden changes in atmospheric pressure,
radiation, and electric shock.
3.Chemical Agents and Drugs-glucose or salt in
hypertonic concentration, poisons, such as arsenic
Cyanide, environmental and air pollutants, insecticides,
and herbicides; industrial and occupational hazards,
such as carbon monoxide and asbestos; social stimuli,
such as alcohol and narcotic drugs; and the ever-
increasing variety of therapeutic drugs.
04/06/2021 305
 4. Infectious Agents- range from the submicroscopic viruses
to the large tapeworms,
5. Immunologic Reactions- Eg. anaphylactic reaction to a
foreign protein or a drug and autoimmune diseases
6.Genetic Derangements – like inborn errors of metabolism
arising from enzymatic abnormalities, usually an enzyme
lack,
7.Nutritional Imbalances- both deficiency and excess, Eg.
PEM and obesity

04/06/2021 306
Examples of causes of cellular injury and their
mode of action

04/06/2021 307
 HYPOXIC INJURY

Cerebral infarction Myocardial infarction

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Renal atrophy 308
INFECTIOUS DISEASE

Tuberculosis Actinomycosis

Candidiasis Primary Herpes

04/06/2021 309
 PHYSICAL INJURY

Thermal Burn Traumatic ulcer

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 CHEMICAL/DRUG INJURY

Gingival Asprin Burn

Hyperplasia
04/06/2021 311
GENETIC DERANGEMENTS

Cancer

Down's Syndrome

04/06/2021 312
Mechanism of cell injury

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04/06/2021 314
Mechanisms of cellular injury. Different agents can injure the various structural and
functional components of the cell. Some cells with specific function are selectively prone
to certain types of injury.
04/06/2021 315
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 Mechanisms of Cell Injury
• Complex process
• Some relevant principles exist;
-The cellular response to injurious stimuli
depends on the type of injury, its duration, and
its severity
-The consequences of cell injury depend on the
type, state, and adaptability of the injured cell.
-The cell’s nutritional and hormonal status and
its metabolic needs are important in its
response to injury.
04/06/2021 317
MECHANISMS OF CELL INJURY
The most important targets of injurious stimuli are
(1) mitochondria, the sites of ATP generation
(2) cell membranes, on which the ionic and osmotic
homeostasis of the cell and its organelles depends
(3) protein synthesis
(4) the cytoskeleton and
(5) the genetic apparatus of the cell.

04/06/2021 318
04/06/2021 319
Depletion of ATP

-frequently associated with both hypoxic and chemical

(toxic) injury
-functions like membrane transport, protein synthesis,
lipogenesis, and the deacylation–reacylation need ATP
-ATP can be produced by oxidative phosphorylation or
glycolytic pathway
-ATP depletion to 5-10% of normal levels has effect on
many critical cellular systems

04/06/2021 320
• Effects of ATP depletion
-plasma membrane energy-dependent sodium pump is
reduced
-Cellular energy metabolism is altered
-Failure of the Ca2+ pump leads to influx of Ca2+,
-reduction in protein synthesis
-Protein misfolding

04/06/2021 321
 HYPOXIA - ISCHEMIA MODEL
Blood Clot
Impaired function of the
plasma membrane
ATP-dependent
 O2
Na+ pump

 Oxidative ATP  Glycolysis

Phosphorylation
Detachment of
ribosomes

04/06/2021 322
 HYPOXIA - ISCHEMIA MODEL
Impaired function of the
plasma membrane Na+ influx
ATP-dependent Ca++ influx  H2O influx
Na+ pump K+ efflux

Cellular swelling
Membrane blebs
and loss of villi
ER swelling

04/06/2021 323
04/06/2021 324
HYPOXIA - ISCHEMIA MODEL
 Glycolysis pH Chromatin
Clumping
Detachment  Glycogen
of ribosomes Stores

 Protein
Synthesis

Lipid
Deposition
04/06/2021 325
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 Mitochondrial damage
• Mitochondria can be damaged by increases of cytosolic Ca2+, by
oxidative stress, by breakdown of phospholipids ,lipid breakdown
products
• Damage results in formation of high conductance channels
which if persistent can affect proton motive force or potential
• Leakage of Cytochrome C which is an integral component of the
electron transport chain to cytosol
• Cytochrome C can trigger apoptotic pathways

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 Influx of intracellular calcium and loss of
calcium homeostasis
• Low levels of intracellular calcium is mainatained by
membrane-associated, energy-dependent Ca2+ Mg2+-
ATPases
• Ischemia and certain toxins calcium from
mitochondria and ER
• Increased calcium activates enzymes including ATPase
,protease ,endonucleases

04/06/2021 329
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Ca++ INDUCED CELL INJURY
Ca++
Ca++ Ca++
Cytoplasmic ionic Ca++

ATPase Phospholipase Protease Endonuclease

ATP Phospholipids Protein DNA

Disruption Damage

04/06/2021 331
 Accumulation of oxygen-derived free
radicals (oxidative stress)
• Small amounts of partially reduced reactive oxygen forms are produced as
an unavoidable byproduct of mitochondrial respiration during energy
generation
• Some of these free radicals can damage lipids, proteins and nucleic acids
• They are referred to as reactive oxygen species
• Cells have mechanism to prevent injury caused by ROS
• An imbalance between ROS generating and defence mechanism result in
oxidative stress
• Free radicals are chemical species that have a single unpaired electron in an
outer orbit

04/06/2021 332
• Free radicals may be initiated within cells in several ways
-Absorption of radiant energy (e.g., ultraviolet light, x-rays).
-Enzymatic metabolism of exogenous chemicals or drugs (e.g., carbon
tetrachloride [CCl4] can generate CCl3)
-The reduction-oxidation reactions that occur during normal metabolic
processes e.g. superoxide anion radical (O2-), hydrogen peroxide(H 2O2),
and hydroxyl ions (OH)
-Transition metals such as iron and copper donate or accept free electrons
during intracellular reactions
-Nitric oxide (NO), an important chemical mediator generated by endothelial
cells, macrophages. . . It can also be converted into peroxynitrite anion
(ONOO-) as well as NO2 and NO3-

04/06/2021 333
Three most important effects
• Lipid peroxidation of membranes
-in the presence of oxygen can cause
peroxidation of lipids within plasma and
organellar membranes
-lipid–free radical interactions yield peroxides,
which are themselves unstable and reactive,
and an autocatalytic chain reaction ensues
(called propagation)

04/06/2021 334
• Oxidative modification of proteins
-promote oxidation of amino acid residue side
chains, formation of protein-protein cross-
linkages and oxidation of the protein backbone,
resulting in protein fragmentation
• Lesions in DNA
-Reactions with thymine in nuclear and
mitochondrial DNA produce single-stranded breaks
in DNA

04/06/2021 335
Free Radicals
Damaging effects
Membrane
Peroxidation of lipids Damage

protein damage Ion Pump

Damage

Impaired
DNA damage Protein Synthesis

Ca Influx
04/06/2021
Mitochondrial damage 336
Into cell
Defence against free radicals

04/06/2021 337
 Defence against free radicals
-free radicals are inherently unstable and generally decay spontaneously
-there are enzymatic and non-enzymatic systems of inactivation
• Antioxidants either block the initiation of free radical formation or
inactivate (e.g., scavenge) free radicals and terminate radical damage
e.g. Vitamins E ,A ,ascorbic acid, glutathione
• Binding of the ions like iron and copper (which generate ROS) to storage
and transport proteins (e.g., transferrin, ferritin, lactoferrin, and
ceruloplasmin), thereby minimizing OH formation

04/06/2021 338
• Enzymes
-Catalase, present in peroxisomes,
which decomposes
H2O2 (2 H2O2 O2 + 2 H2O)
-Superoxide dismutases are found in many
cell types and convert superoxide to H2O2
(2 O2- + 2 H H2O2 + O2)
-Glutathione peroxidase also protects against injury by
catalyzing free radical breakdown

04/06/2021 339
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 Defects in membrane permeability

-may affect mitochondria ,plasma membrane and other

cellular membranes
-results from ATP depletion and calcium modulated
action of phospholipases
-plasma membrane may also be directly injured by
bacterial toxins, viral proteins, lytic complement
components, and physical and chemical agents

04/06/2021 341
Biochemical mechanisms of membrane damage
• Mitochondrial dysfunction
• Loss of membrane phospholipids
• Cytoskeletal abnormalities
• Reactive oxygen species
• Lipid breakdown products

04/06/2021 342
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04/06/2021 344
Cytoskeletal abnormalities
• . Cytoskeletal filaments serve as anchors connecting the
plasma membrane to the cell interior.
• Activation of proteases by increased cytosolic calcium
may cause damage to elements of the cytoskeleton

04/06/2021 345
Cont’d…
• CYTOSKELETAL ABNORMALITIES
• Cytoskeletal abnormalities may be reflected by
• Defect in cell locomotion and intracellular organelle movement
• Intracellular accumulations of fibrillar material
• Cytoskeleton consists of
• Thin filaments- actin, myosin
• Microtubules
• Intermediate filaments
• Keratin – epithelial cells
• Neurofilaments- neurons
• Desmin – muscle cells
• Vimentin- connective tissue cells
• Glial filaments - astrocytes

04/06/2021 346
Protein synthesis

04/06/2021 347
DNA damage or loss

04/06/2021 348
The types of DNA damage include:

• strand breaks

• base alterations

• cross-linking

04/06/2021 349
Morphology of cell injury
• Cells undergo sequential biochemical and morphologic
changes
• Initially molecular and biochemical changes then
morphologic changes
• Histochemical and ultrastructural changes may be seen in
minutes or hours
• Light microscopic and gross changes takes considerably
longer time
• E.g. light morphologic changes of cell death in the
myocardium occur after 4-12hrs of total ischemia but
process starts after 20-60 min

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04/06/2021 351
Reversible vs. irreversible cell injury

• The stage of irreversible cell injury /the “point of no

return,” is still largely undetermined .
• Two phenomena consistently characterize irreversibility.
-1. the inability to reverse mitochondrial
dysfunction (lack of oxidative phosphorylation
and ATP generation) even after resolution of the
original injury.
-2. the development of profound disturbances in
membrane function.

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Morphology of Cell Injury and Necrosis

All stresses and noxious influences exert their effects first at

the molecular or biochemical level

There is a time lag between the stress and the morphologic

changes of cell injury or death; the duration of this delay
may vary with the sensitivity of the methods used to detect
these changes.

04/06/2021 353
• With histochemical or ultrastructural techniques, changes
may be seen in minutes to hours after ischemic injury;

• however, it may take considerably longer (hours to days)

before changes can be seen by light microscopy or on gross

examination.

04/06/2021 354
Morphology of reversible injury
• Cellular swelling is the first manifestation of almost all
forms of injury to cells.
• It is a difficult morphologic change to appreciate with the
light microscope;
• When it affects many cells in an organ, it causes some
pallor, increased turgor, and increase in weight of the organ.
• On microscopic examination, small clear vacuoles may be
seen within the cytoplasm; these represent distended and
pinched-off segments of the endoplasmic reticulum.

04/06/2021 355
•This pattern of nonlethal injury is sometimes called
hydropic change or vacuolar degeneration.
• Swelling of cells is reversible.
•The ultrastructural changes of reversible cell injury include:
1. plasma membrane alterations, such as blebbing, blunting,
and distortion of microvilli; and loosening of intercellular
attachments
2. mitochondrial changes, including swelling, and the
appearance of small phospholipid-rich amorphous densities
3. dilation of the endoplasmic reticulum, with detachment and
disaggregation of polysomes
4. nuclear alterations, with disaggregation of granular and
fibrillar elements.
Chronic non-lethal cell injury manifest morphologically as fatty
changes.
04/06/2021
Eg. Fatty liver in alcoholics 356
Cellular swelling

04/06/2021 357
Fatty Change
“Fatty Liver”

04/06/2021 358
Morphology of Irreversible
injury/necrosis
• Necrosis refers to a spectrum of morphologic changes that
follow cell death in living tissue, largely resulting from the
progressive degradative action of enzymes on the lethally
injured Cell
• The morphologic appearance of necrosis is the result of
denaturation of intracellular proteins and enzymatic
digestion of the cell.
• The enzymes are derived either from the lysosomes of the
dead cells themselves, in which case the enzymatic digestion
is referred to as autolysis, or from the lysosomes of immigrant
leukocytes, during inflammatory reactions/heterolysis/.

04/06/2021 359
• Nuclear changes appear in the form of one of three
patterns, all due to nonspecific break down of DNA.
• pyknosis- characterized by nuclear shrinkage and increased
basophilia. Here the DNA apparently condenses into a solid,
shrunken basophilic mass.
• Karyorrhexis-the pyknotic or partially pyknotic nucleus
undergoes fragmentation. With the passage of time (a day
or two), the nucleus in the necrotic cell totally disappears.
• Karyolysis-The basophilia of the chromatin fades due to
dissolution of the nucleus and fragments.This change
presumably reflects DNase activity.
04/06/2021 360
 Necrosis

Pyknosis Karyolysis

Normal Karyorrhexis

04/06/2021 361
• Morphology -- Necrotic cells show increased
eosinophilia attributable in part to loss of the normal
basophilia imparted by the RNA in the cytoplasm and
in part to the increased binding of eosin to denatured
intracytoplasmic proteins .
• The necrotic cell may have a more glassy homogeneous appearance
than that of normal cells, mainly as a result of the loss of glycogen
particles.

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04/06/2021 365
04/06/2021 366
04/06/2021 367
Types of necrosis

• Coaggulative necrosis –when denaturation is

the predominant feature.
-Basic outline of the coagulated cells is preserved
- intracellular acidosis denatures both structural
proteins and enzymes and so blocks the proteolysis
of the cell,
• Is characterstic of hypoxic cell death of all tissues
except the brain
- myocardial infarct is an excellent example

04/06/2021 368
Renal Infarction - Coagulative

04/06/2021 369
Coagulative necrosis
Ischemic injury to the
glomerulus kidney

Outline of the
glomerulus and tubules
are still visible, despite
tubules the cell being dead

tubules

04/06/2021 370
 B

• A, Normal myocardium. B, Myocardium with coagulation necrosis having

inflammatory rsponse

04/06/2021 371
Liquefactive necrosis – in case of dominant enzyme
digestion,
- is characteristic of focal bacterial or, occasionally, fungal
infections followed by accumulation of inflammatory cells,
-If the process was initiated by acute inflammation,
accumulation of dead inflammatory cells form creamy
yellow material/pus
-The most common types of damage leading to the
liquefactive pattern are necrosis in the brain owing to arterial
occlusion (cerebral infarction and necrosis caused by bacterial
infections).

04/06/2021 372
Stroke- Liquefactive necrosis

04/06/2021 373
Liver abscess: Liquefactive necrosis

04/06/2021 374
• Coagulative and liquefactive necrosis. A, Kidney infarct;
• B, A focus of liquefactive necrosis in the kidney caused by fungal infection..

04/06/2021 375
Liquefactive necrosis
Liquefaction necrosis
of a cerebral infarct

Semi-fluid mass of
protein and no
cellular structure

04/06/2021 376
• Caseous necrosis- a distinctive form of coagulative
necrosis encountered most often in foci of tuberculous
infection
-amorphous granular debris enclosed within a distinctive
inflammatory border known as a granulomatous reaction

04/06/2021 377
 Extensive
Caseous necrosis
Tuberculosis

04/06/2021 378
FIGURE 1–20 A tuberculous lung with a large area of caseous
necrosis. The caseous debris is yellow-white and cheesy.
04/06/2021 379
Caseous necrosis

04/06/2021 380
Caseous Necrosis
Caseous necrosis of a
kidney infected with
Mycobacterium
tubelculosis

Necrotic area is
homogenously
pink
The necrotic area is
homogenously pink
and no renal
architecture
04/06/2021 381
• Gangrenous necrosis- is not a distinctive
pattern of cell death,
-coagulative necrosis due to ischemia is modified by
the liquefactive action of the bacteria and the
attracted leukocytes (so-called wet gangrene).

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Gangrene – With esp. infection

• -It is usually applied to a limb, generally the lower leg, that

has lost its blood supply and has undergone coagulative
necrosis involving multiple tissue layers.
-When bacterial infection is superimposed, coagulative
necrosis is modified by the liquefactive action of the
bacteria and the attracted leukocytes (so-called wet
gangrene).

04/06/2021 383
 Gangrene

• Gangrene is applied when considerable amount of tissue

under goes necrosis.
• There are three types of gangrene :
• 1.dry gangrene usually due to interference of
arterial blood supply but patent venues system.
• 2.moist gangrene or wet gangrene due to
interference of venous return and also usually
associated with supper imposed infection.
• 3.gas gangrene is a special type of gangrene which is due to
infection of devitalized tissue by clostradial spp. Produce gas bubbles. And
it is fatal.
04/06/2021 384
Gangrene - Amputated Diabetic foot

04/06/2021 385
Gangrenous
necrosis

04/06/2021 386
 Hemmorrhagic necrosis

This pattern is seen particularly when cell

death is due to blockage of the venous
drainage of a tissue, leading to massive
congestion by blood and to subsequent
arterial failure of perfusion

04/06/2021 387
• Fat necrosis – focal areas of fat destruction occurring as
a result of release of activated pancreatic lipases into the
substance of the pancreas and the peritoneal cavity →Acute
pancreatitis which is an abdominal emergency
• released fatty acids combine with calcium to produce grossly
visible chalky white areas (fat saponification)

04/06/2021 388
Fat necrosis Fat necrosis & saponification

04/06/2021 389
FIGURE 1–21 Foci of fat necrosis with saponification in the
mesentery. The areas of white chalky deposits represent calcium
soap formation at sites of lipid breakdown.
04/06/2021 390
Cont’d…
• Ultimately most necrotic cells disappear by
• Enzymatic digestion and fragmentation
• Phagocytosis of the particulate debris by leukocytes.
• If necrotic cells and cellular debris are not promptly
destroyed and reabsorbed, they tend to attract
calcium salts. This phenomenon is called dystrophic
calcification.

04/06/2021 391
 Apoptosis
• It is a death of single cell in cluster of other cells.
• is a pathway of cell death that is induced by a tightly regulated suicide
program in which cells destined to die activate enzymes capable of
degrading the cells' own nuclear DNA and nuclear and cytoplasmic
proteins.
• The plasma membrane of the apoptotic cell remains intact, but the
membrane is altered in such a way that the cell and its fragments
become avid targets for phagocytes. The dead cell is rapidly cleared
before its contents have leaked out, and therefore cell death by this
pathway does not elicit an inflammatory reaction in the host.

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• apoptosis differs from necrosis, which is characterized by
loss of membrane integrity, enzymatic digestion of cells,
leakage of cellular contents, and frequently a host reaction.
• Usually physiologic,which occurs as programmed cell death.
• It does not follow inflammation.
• apoptosis and necrosis sometimes coexist,
• Examples
• During embryogenesis
• During menstruation

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 Apoptosis in Physiologic Situations

• Death by apoptosis is a normal phenomenon that

serves to eliminate cells that are no longer needed,
as, for example, during development, and to maintain
a steady number of various cell populations in
tissues.  

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 Apoptosis in Physiologic Situations…

• Hormone-dependent involution in the adult:

• such as endometrial cell breakdown during the menstrual cycle,
ovarian follicular atresia in the menopause, the regression of the
lactating breast after weaning, and prostatic atrophy after
castration.
•Death of host cells that have served their useful purpose,
such as neutrophils in an acute inflammatory response, and
lymphocytes at the end of an immune response
•Elimination of potentially harmful self-reactive lymphocytes,
either before or after they have completed their maturation.
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Apoptosis in Pathologic Conditions
• DNA damage.
Radiation,cytotoxic anticancer drugs, extremes of temperature,
and even hypoxia can damage DNA, either directly or via
production of free radicals.
If repair mechanisms can’t cope with the injury, the cell triggers
intrinsic mechanisms that induce apoptosis.
• Accumulation of misfolded proteins.
Improperly folded proteins may arise because of mutations in the
genes encoding these proteins or because of extrinsic factors, such
as damage caused by free radicals. Excessive accumulation of
these proteins in the ER leads to a condition called ER stress, which
culminates in apoptotic death of cells.

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• The fundamental event in apoptosis is the activation of
enzymes called caspases (so named because they are
cysteine proteases that cleave proteins after aspartic
residues). Activated caspases cleave numerous targets,
culminating in activation of nucleases that degrade
DNA and other enzymes that presumably destroy
nucleoproteins and cytokeletal proteins. The activation
of caspases depends on a finely tuned balance
between pro- and anti-apoptotic molecular pathways.
• Two distinct pathways converge on caspase activation,
called the mitochondrial pathway and the death
receptor pathway.

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 The Mitochondrial (Intrinsic) Pathway of
Apoptosis
• is responsible for most situations of apoptosis.
• Mitochondria contain several proteins that are capable of
inducing apoptosis; these proteins include cytochrome c and
antagonists of endogenous cytosolic inhibitors of apoptosis.
• When cells are deprived of growth factors and trophic
hormones, or are exposed to agents that damage DNA, or
accumulate unacceptable amounts of misfolded proteins, a
group of sensors is activated.

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• Some of these sensors, which are members of the Bcl-2
family, in turn activate two pro-apoptotic members of the
family called Bax and Bak, which dimerize, insert into the
mitochondrial membrane, and form channels through
which cytochrome c and other mitochondrial proteins
escape into the cytosol.
• Cytochrome c, together with some cofactors, activates
caspase-9, while other proteins block the activities of
caspase antagonists that function as physiologic inhibitors
of apoptosis. The net result is the activation of the caspase
cascade, ultimately leading to nuclear fragmentation. If
cells are exposed to growth factors and other survival.

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• signals, they synthesize antiapoptotic members of
the Bcl-2 family, the two main ones of which are
Bcl-2 itself and Bcl-xL. These proteins antagonize
Bax and Bak, and thus limit the escape of
mitochondrial pro-apoptotic proteins

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 The Death Receptor (Extrinsic) Pathway of Apoptosis
• Many cells express surface molecules, called death receptors, that
trigger apoptosis. Most of these are members of the tumor necrosis
factor (TNF) receptor family that contain in their cytoplasmic regions a
conserved "death domain," so named because it mediates interaction
with other proteins. The prototypic death receptors are the type I TNF
receptor and Fas (CD95).
• Fas-ligand (FasL) is a membrane protein expressed mainly on activated
T lymphocytes. When these T cells recognize Fas-expressing targets,
Fas molecules are cross-linked by the FasL and they bind adapter
proteins, which in turn bind caspase-8. Clustering of many caspase
molecules leads to their activation, thus initiating the caspase cascade.

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MORPHOLOGY OF APOPTOSIS

Chromatin condensation
Progressive cell shrinkage
Plasma membrane blebbing
Apoptotic bodies
Phagocytosis
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 Features of Necrosis and Apoptosis
Features Necrosis Apoptosis
Cell size Reduced(shrinkage)
Enlarged(swelling)

Nucleus Pyknosis-karyorrhexis- Fragmentationin to

karyolysys nucleosome size fragments

Plasma membrane Disrupted Intact; altered structure,

especially orientation of
lipids

Cell contents Enzymatic digestion, may Intact; may be released in

leak out of cell apoptotic bodies

Adjacent inflammation Frequent No

Physiologic/pathologic role Invariably Often physiologic, means
pathologic(culminatiion of of eliminating unwanted
irreversible cell injury) cells; may be pathologic
after some form of cell
injury,esp. DNA damage
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Subcellular Responses to Injury
LYSOSOMAL CATABOLISM
• Primary lysosomes are membrane-bound intracellular
organelles that contain a variety of hydrolytic enzymes,
• These enzymes are synthesized in the rough endoplasmic
reticulum and then packaged into vesicles in the Golgi
apparatus.
• Primary lysosomes fuse with membrane-bound vacuoles
that contain material to be digested,forming secondary
lysosomes or phagolysosomes.
• Lysosomes are involved in the breakdown of phagocytosed
material
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431 .
• Heterophagy- is the process of lysosomal digestion of
materials ingested from the extracellular environment.
• Extracellular materials are taken up by cells through the
general process of endocytosis.
• Uptake of particulate matter is known as phagocytosis;
• uptake of soluble smaller macromolecules is called
pinocytosis.
• Endosomes or phagosomes), fuse with lysosomes to form
phagolysosomes.
• Heterophagy is most common in the “professional”
phagocytes, such as neutrophils and macrophages
• heterophagocytosis include the uptake and digestion of
bacteria by neutrophils and the removal of apoptotic cells by
macrophages.
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• Autophagy- refers to lysosomal digestion of the cell’s own
components.
• intracellular organelles and portions of cytosol are first
sequestered from the cytoplasm in an autophagic vacuole
formed from ribosome-free regions of the rough endoplasmic
reticulum.
• The vacuole fuses with lysosomes or Golgi elements to form
an autophagolysosome.
• Autophagy is a common phenomenon involved in the removal
of damaged organelles during cell injury and the cellular
remodeling of differentiation, and it is particularly pronounced
in cells undergoing atrophy induced by nutrient deprivation or
hormonal involution.

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Autophagy

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• The enzymes in lysosomes are capable of degrading most
proteins and carbohydrates, but some lipids remain
undigested.
• Lipofuscin pigment granules represent undigested material
derived from intracellular lipid peroxidation.
• Hereditary lysosomal storage disorders, caused by
deficiencies of enzymes that degrade various
macromolecules, result in the accumulation of abnormal
amounts of these compounds
in the lysosomes of cells all over the body,

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INDUCTION (HYPERTROPHY) OF SMOOTH
ENDOPLASMIC RETICULUM
• The smooth ER is involved in the metabolism of various
chemicals.
• cells exposed to these chemicals show hypertrophy of the
ER as an adaptive response that may have important
functional consequences.
• Protracted use of barbiturates leads to a state of tolerance,
with a decrease in the effects of the drug due to increased
volume (hypertrophy) of the smooth ER of hepatocytes,
which metabolizes the drug

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 SER Induction

Hypertro
phy of Normal ER ER hyperplasia
Smooth
Endoplas
mic

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MITOCHONDRIAL ALTERATIONS
• mitochondrial dysfunction plays an important role in cell injury
and apoptosis.
• various alterations in the number, size, and shape of
mitochondria occur in some pathologic conditions.
• Mitochondria may assume extremely large and abnormal shapes
(megamitochondria), like in alcoholic liver disease and in certain
nutritional deficiencies.
• In inherited metabolic diseases of skeletal muscle, the
mitochondrial myopathies, defects in mitochondrial metabolism
are associated with increased numbers of mitochondria that are
often unusually large, have abnormal cristae, and contain
crystalloids

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Mitochondrial Hyperplasia

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 Megamitochondria

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INTRACELLULAR ACCUMULATIONS

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INTRACELLULAR
ACCUMULATIONS
• Normal constituents - H20, lipids, proteins,
carbohydrate
• Abnormal substances- endogenous
or exogenous
• Pigments
• Calcium

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INTRACELLULAR ACCUMULATIONS

Steatosis

Pigment storage

Cholestrol storage

Protein storage

Glycogen storage
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•most accumulations are attributable to three types of
abnormalities.
•1. A normal endogenous substance is produced at a normal or
increased rate, but the rate of metabolism is inadequate to
remove it. Eg.fatty change in the liver
•2. A normal or abnormal endogenous substance accumulates
because of genetic or acquired defects in the metabolism,
packaging, transport, or secretion of these substances. Eg.
Storage diseases
•3.An abnormal exogenous substance is deposited and
accumulates because the cell has neither the enzymatic
machinery to degrade the substance nor the ability to
transport it to other sites. Eg.Accumulations of carbon
particles.
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MECHANISMS OF INTRACELULAR
ACCUMULATIONS

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• LIPIDS
-All major classes of lipids can accumulate in cells:
triglycerides,cholesterol/cholesterol esters, and
phospholipids.
-In industrialized nations, by far the most common cause
of significant fatty change in the liver (fatty liver) is
alcohol abuse.
Fatty liver

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 Steatosis (Fatty Change)
The terms steatosis and fatty change describe abnormal
accumulations of triglycerides within parenchymal cells.
 Fatty change is often seen in the liver because it is the
major organ involved in fat metabolism, but it also occurs
in heart muscle, and kidney.
 The cause of steatosis include toxins, protein malnutrition,
diabetes mellitus, obesity, and anoxia.
In industrialized nations, by far the most common cause of
significant fatty change in the liver (fatty liver) is alcohol
abuse & DM asstd with obesity.

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 Steatosis (Fatty Change)…
• Different mechanisms account for triglyceride accumulation
in the liver.
• Free fatty acids from adipose tissue or ingested food are
normally transported into hepatocytes.
• In the liver, they are esterified to triglycerides, converted
into cholesterol or phospholipids, or oxidized to ketone
bodies.
• Egress of the triglycerides from the hepatocytes requires the
formation of complexes with apoproteins to form
lipoproteins, which are able to enter the circulation. Defects
at any step from fatty acid entry to lipoprotein exit result in
excess accumulation TG in the liver causing fatty liver.
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• Hepatotoxins (e.g., alcohol) alter mitochondrial and SER
function and thus inhibit fatty acid oxidation.
• CCl4 and protein malnutrition decrease the synthesis of
apoproteins.
• Anoxia inhibits fatty acid oxidation and starvation
increases fatty acid mobilization from peripheral stores.

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Steatosis (Fatty Change)…

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Other Lipid accumulation
• Cholesterol and Cholesterol Esters
• 1. Atherosclerosis.
• A vascular intimal accumulation of cholesterol esters
• 2. Extracellular cholesterol esters (Cholesterol clefts)
may crystallize in the shape of long needles (clefts) in
tissue sections.
• 3. Xanthomas.
• Clusters of foamy cells are found in the subepithelial
connective tissue of the skin and in tendons.

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Cont’d…
4. Inflammation and necrosis.
 Foamy macrophages are found at sites of inflammation, owing to
phagocytosis of cholesterol from the membranes of injured cells.
5. Cholesterolosis.
 Focal accumulations of cholesterol-laden macrophages in the
lamina propria of the gallbladder
6. Niemann-Pick disease, type C.
 An enzyme involved in cholesterol trafficking is mutated, and
hence cholesterol accumulates in multiple organs
7. Guacher’s disease: Sphingolipidoses in MNP cells

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CHOLESTEROL IN VESSELS

Atherosclerosis Cholesterol thrombus

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PROTEINS
• Intracellular accumulations of proteins usually appear as
rounded, eosinophilic droplets, vacuoles, or aggregates in
the cytoplasm.
• Aggregation of abnormal proteins may deposit in tissues as
intracellular, extracellular or both and interfere with normal
functions.Eg .Amyloidosis

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GLYCOGEN
• Excessive intracellular deposits of glycogen are seen in
patients with an abnormality in either glucose or glycogen
metabolism.
• Glycogen also accumulates within the cells in a group of
closely related disorders referred to as the glycogen storage
diseases, or glycogenoses

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Endogenous Pigments
-Lipofuscin is an insoluble pigment, also known as
lipochrome and wear-and-tear or aging pigment.
- Lipofuscin is composed of polymers of lipids and
phospholipids complexed with protein, suggesting that it
is derived through lipid peroxidation of polyunsaturated
lipids of subcellular membranes.
- It is seen in cells undergoing slow, regressive changes
and is particularly prominent in the liver and heart of aging
patients or patients with severe malnutrition and cancer
cachexia.

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 Pigment cont…
Melanin, derived from the Greek (melas = black), is an
endogenous, non-hemoglobin-derived, brown-black
pigment formed when the enzyme tyrosinase catalyzes the
oxidation of tyrosine to dihydroxyphenyalanine in
melanocytes. It is synthesized exclusively by melanocytes
located in the epidermis and acts as a screen against
harmful ultraviolet radiation.
Hemosiderin is a hemoglobin-derived, golden yellow-to-
brown, granular or crystalline pigment in which form iron is
stored in cells. Iron is normally stored within cells in
association with the protein apoferritin, forming ferritin
micelles.
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 Bilirubin

• Bilirubin is the normal major pigment found in bile.

• It is derived from hemoglobin but contains no iron. Its
normal formation and excretion are vital to health, and
jaundice is a common clinical disorder caused by
excesses of this pigment within cells and tissues .
• May be caused by:
• Hemolytic anemia
• Biliary obstraction
• Hepatocellular disease

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Calcifications

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 ! ! !
C H
M U
S O
K U
A N
TH

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