Course 1- recap

Properties Primary cells

What is histopathology? The microscopic study of diseased tissue

The most used apparatus in the histology labs to cut Microtome/Cryotome
tissue sections
What Haematoxylin-eosin staining is showing?
Hematoxylin shows the ribosomes, chromatin (genetic
material) within the nucleus a deep purple-bluish color.
Eosin gives cytoplasms a pink color

Haematoxylin-eosin staining of a glomerulus of a

kidney.
 Course 1- recap
Properties
Staining is the process for coloring tissues by using dyes.
What contains a dye molecule?
Special techniques used in Histopathology
Immunostains are composed of specific antibodies that
should only bind to the corresponding antibody in the
tissues.
There are two main methods of immunodetection:
The commonest used chromogen which results in
characteristic ‘brown’ staining.
Primary cells
A dye molecule has two domains: the chromogen provides
the color and the auxochrome makes possible the binding to
the tissue. Chromophore is the molecular structure of the
chromogen that absorbs a particular light wavelength;
- the direct method
- the indirect method involves the primary antibody (P) binding to a
labelled secondary antibody (SA); The secondary antibody is
labelled with an enzyme (E) (chromogen), which converts a
substrate to a coloured compound (Co) which allows visualisation
of the reaction in the tissues.
is horseradish peroxidase,
Ki-67 staining
 Course 2- recap
Properties Primary cells

What is Edema?
Disorder that perturb cardiovascular, renal, or hepatic
function are often marked by the accumulation of fluid in
tissues (edema) or body cavities (effusions).

Hemostasis is the process by which blood clots form at sites of

What is Hemostasis? What are the stages of hemostasis?
vascular injury.
1. Arteriolar vasoconstriction 2. Primary hemostasis. 3. Secondary
hemostasis 4. Clot stabilization and resorption.

Hemorrhagic disorders associated with abnormal bleeding myocardial infarction complicated by rupture of
inevitably stem from primary or secondary defects in vessel the aorta or the heart.
walls, platelets, or coagulation factors, all of which must
function properly to ensure hemostasis.
Hemorrhagic disorders examples?

What are the primary abnormalities that lead to

thrombosis?
(1) endothelial injury;
(2) stasis or turbulent blood flow;
(3) hypercoagulability of the blood (the so-called Virchow
triad).
 Course 2- recap
Properties Primary cells

Name a cause of inflammation

Infections
Tissue necrosis
Foreign bodies
Immune reactions

Acute
What are the major types of inflammation?
Chronic
What are the major components of Acute inflammation ?
I. Dilation of small vessels leading to an increase in blood flow;

II. Increased permeability of the microvasculature enabling

plasma proteins and leukocytes to leave the circulation;

III. Emigration of leukocytes from the microcirculation, their

accumulation in the focus of injury and their activation to
eliminate the offending agent.
What is chronic inflammation?
Chronic inflammation is a response of prolonged duration
(weeks or months) in which inflammation, tissue injury and
attempts at repair coexist in varying combinations.
 Adaptations of cellular
growth and differentiation:
hypertophy, hyperplasia,
atrophy, metaplasia and

dysplasia
 Adaptations are reversible changes in the size, number, phenotype, metabolic
activity, or functions of cells in response to changes in their environment (Oakes,
2020).
• Physiologic adaptations

• Pathologic adaptations

Adaptations GROWTH:
- hypertrophy;
- hyperplasia;
- atrophy.

DIFFERENTIATION:
- metaplasia
- dysplasia
Hypertrophy
Hypertrophy is an increase in the size of cells that results in an increase in the size of the affected organ.

• Pathologic hypertrophy:
- enlargement of the heart in response to pressure overload
- increased workload of the skeletal muscle

muscle cells respond by

synthesizing more protein and
increasing the number of
myofilaments per cell
• Physiologic hypertrophy:
- hormone-induced enlargement of an organ that results
mainly from hypertrophy of smooth muscle fibers
(e.g. estrogen through estrogen receptors)
Hypertrophy – physiologic adaptations
Mechanisms of hypertrophy
- Mechanical sensors appear to be
the major triggers for physiologic
hypertrophy, and agonists and
growth factors may be more
important in pathologic states.
- ANF (Atrial natriuretic factor),
GATA4 (transcription factor that
binds to DNA sequence GATA),
IGF1 (insulin-like growth factor)
NFAT (nuclear factor–activated T
Cells), MEF2 (myocardial
enhancing factor 2).

Biochemical mechanisms of myocardial hypertrophy.

Hypertrophy – pathologic adaptations
Normal heart vs. cardiac hypertrophy. a Cross section of a
normal heart in a perinatal death: the right ventricular free wall
thickness is 3.5 mm, the left ventricular is 5 mm, and the
septum 5.5 mm. b Cross section of a normal adult heart: the
right ventricular free wall thickness is 2 mm, the left ventricular
is 12 mm, and the septum 13 mm. c Cross section of
hypertrophic heart in an adult: the right ventricular free wall
thickness is 7 mm, the left ventricular is 21 mm, and the septum
22 mm. d Histology of a showing hypercellularity which is
normal for a perinatal myocardium (high number of cardiac
myocyte/myocardial area) (bar = 100 micron). e Histology
of b with diameter of cardiac myocytes within normal values
(mean diameter 12 micron) (bar = 100 micron). f Histology
of c with cardiac myocyte hypertrophy (mean diameter 20
micron) (bar = 100 micron)

Basso, C., Michaud, K., d’Amati, G. et al. Cardiac hypertrophy

at autopsy. Virchows Arch 479, 79–94 (2021).
https://doi.org/10.1007/s00428-021-03038-0
Hyperplasia
Hyperplasia is an increase in the number of cells in an organ or tissue in response to a stimulus.
• Pathologic hyperplasia:
- excessive/inappropriate action of hormones
or growth factors acting on target cells
(e.g disturbed balance between estrogen and progesterone
leads to abnormal uterine bleeding)
- benign prostatic hyperplasia
- in viral infections (papillomaviruses): mucosal lesions
(masses of hypeplastic epithelium)
• Physiologic hyperplasia:
- action of hormones or growth factors
(e.g proliferation of glandular epithelium of female breasts
at puberty and pregnancy)
- compensatory: liver regeneration
- bone marrow in response to deficiency of blood cells
Hyperplasia – physiologic

Hormonal hyperplasia is well illustrated by the proliferation of the glandular epithelium of the female breast at
puberty and during pregnancy, usually accompanied by enlargement (hypertrophy) of the glandular epithelial cells.
Mechanisms of hyperplasia

- The liver progenitor cells (LPCs) and ductular reaction

(DR):
- (A) Localization of LPCs within the liver;

- (B) dynamics of the differentiation process of DR

cells: During injury, DR starts to emerge; DR cells will
differentiate into hepatocytes through intermediate
steps.
Hyperplasia – pathologic

- the balance between estrogen and progesterone is disturbed,

resulting in absolute or relative increases in the amount of
estrogen, with consequent hyperplasia of the endometrial
glands.
Hyperplasia – pathologic

Benign prostatic hyperplasia is another common example of pathologic hyperplasia, in this case as a response to hormonal
stimulation by androgens. Although these forms of pathologic hyperplasias are abnormal, the process remains controlled
and the hyperplasia can either regress or stabilize if the hormonal stimulation is eliminated.
Atrophy
Atrophy is a reduction in the size of an organ or tissue due to a decrease in cell size and number.

• Pathologic atrophy: • Physiologic atrophy:

- decreased workload (disuse atrophy);
- loss of innervation (denervation atrophy);
- diminished blood supply;
- inadequate nutrition;
- loss of endocrine stimulation;
- pressure.
- during normal development
(e.g notochord, tyroglosal duct undergo atrophy
during fetal development);
- the decrease in the size of uterus after
parturition.
Mechanisms of atrophy
Atrophy - pathologic

H&E stain
Grouped atrophy
  Small muscle fibers: Often rounded
  Pyknotic nuclear clumps: None
Large muscle fibers: Hypertrophy
Metaplasia
Metaplasia is a reversible change in which one differentiated cell type (epithelial or mesenchymal) is
replaced by another cell type.

- the most common epithelial metaplasia is columnar to

squamous
(e.g in the respiratory tract in response to chronic irritation;
Vitamin A deficiency can also induce squamous metaplasia in the
respiratory epithelium and in the cornea)

The influences that predispose to

metaplasia, if persistent, can initiate
malignant transformation in metaplastic
epithelium.

Metaplasia of columnar to squamous epithelium (A)

Schematic diagram; (B) Metaplasia of columnar epithelium
(left) to squamous epithelium (right) in a bronchus.
Metaplasia
- metaplasia from squamous to columnar type:
(e.g Barrett esophagus, in which the esophageal squamous epithelium is replaced by intestinal-like columnar cells under
the influence of refluxed gastric acid)

- connective tissue metaplasia = the formation of cartilage, bone, or adipose cells (mesenchymal tissues) in tissues that
normally do not contain these elements
(e.g myositis ossificans, occasionally occurs after intramuscular hemorrhage)
Mechanisms of metaplasia
Metaplasia does not result from a change in the phenotype of an already differentiated cell type; rather, it results from either
reprogramming of local tissue stem cells or, alternatively, colonization by differentiated cell populations from adjacent sites.

Squamous metaplasia. In tissue columnar cells (for example, in the lung or cervix), external stimuli (for
example, low vaginal pH in the cervix and cigarette smoke in the lung) promote the conversion to metaplastic
squamous cells, which stratify. In the lung, columnar cells are identified by the expression of homeobox
protein Nkx2.1 (NKX2-1), and squamous cells are enriched for p63 and SRY-box 2 (SOX2), similar to
oesophageal basal cells and their marked expression of p63 and SOX2.
Mechanisms of metaplasia

Intestinal metaplasia in the oesophagus. The normal oesophageal squamous epithelial proliferative basal cells (p63+ and
SRY-box 2 (SOX2)+) undergo early and terminal differentiation as they migrate towards the luminal surface. In concert
with acid or bile reflux and pro-inflammatory stimuli (for example, interleukin 6 (IL ‑6)–signal transducer and activator of
transcription 3 (STAT3)), incomplete intestinal metaplasia (presence of columnar cells and goblet cells and absence of
Paneth cells and enteroendocrine cells) appears. 

Giroux and Rustgy, 2017 – Nature reviews

Dysplasia
Dysplasia is a term that literally means “disordered growth.” It is encountered principally in epithelial cells and is
recognized on the basis of several morphologic changes. Dysplastic cells may exhibit considerable pleomorphism and
often contain large hyperchromatic nuclei with a high nuclear-to-cytoplasmic ratio. Dysplastic epithelial surfaces also
typically show architectural disarray and a loss of orderly differentiation.
Mechanisms of dysplasia

Nevertheless, although dysplasia may be a

precursor to malignant transformation, it
does not always progress to cancer. With
removal of inciting causes, even
moderately severe dysplasias may be
completely reversible.

Spasmolytic polypeptide-expressing metaplasia and

gastric intestinal metaplasia. The gastric epithelium
harbours chief cells at the base, underneath acid-producing
parietal cells, progenitor cells (or stem cells) and surface
cells. In the face of Helicobacter pylori infection, there is
parietal cell loss and chronic inflammation. One pathway
results in foveolar hyperplasia and spasmolytic
polypeptide-expressing metaplasia (SPEM), which might
be a precursor to intestinal metaplasia (IM, indicated by the
dashed arrow). Another pathway leads directly to IM. Both
SPEM and IM are precursors to dysplasia and later
adenocarcinoma. Please refer to the main text for a
discussion of cell of origin

Giroux and Rustgy, 2017 – Nature reviews

 Find the right label for the following images:

…
…

Normal cells

…
…