CONNECTIVE TISSUE

LECTURE
CONNECTIVE TISSUE

• Continuum with others tissues

• Originating from mesoderm (mesenchyme)
• Composition:
• cells (fibroblasts and others),
• extracellular matrix (fibers and ground substance)
FUNCTION

• Architectural framework (structure, capsules, stroma, isolation)

• Transport medium (oxygen, nutrients, waste)
• Defense (phagocytosis, inflammatory response, antibodies)
• Storage (fat)
EXTRACELLULAR MATRIX

• Fibers – collagen & elastic

• “Ground substance”
• Resisting compressive and stretching forces
GROUND SUBSTANCE

• Ground substance of connective tissue is a homogeneous semifluid

material that surrounds the cells and fibers
• Is composed of glycosaminoglycans, proteins (proteoglycans and
adhesive glycoproteins), water, and minerals
• Glycosaminoglycans: the commonest are hyaluronate, chondroitin-4-
sulfate, chondroitin-6-sulfate
GROUND SUBSTANCE

• Preferential adhesive glycoproteins:

• Laminin – basal lamina
• Chondronectin – cartilage
• Osteonectin – bone
• Dispersed:
• Fibronectin
• Hyaluronic acid is very important because the viscosity of ground substance
depends on the content of it (gel state). Proteoglycans are covalently linked to
hyaluronic acid forming huge macromolecules called aggrecan aggregates
FIBERS - COLLAGEN

• Most abundant protein in human body (up to 30% dry weight)

• Multiple types: fibril-associated (in skin, tendon, cartilage, bone,
dentin, blood vessels); cross-linked networks (in all basement
membranes)
• Consists of tropocollagen subunits, which is composed of three alpha-
chains wrapped around one another in helical configuration
FIBERS - COLLAGEN

• Most common amino acids: glycine (every third), proline,

hydroxyproline, hydroxylysine
• So far around 20 types of collagen have been categorized
• H+E staining – pink
• Electronogram – gap regions
FIBERS - COLLAGEN
FIBERS - COLLAGEN

H&E Trichrome
FIBERS - COLLAGEN
FIBERS – COLLAGEN
FIBERS – COLLAGEN ASSEMBLY
FIBERS - COLLAGEN
EHLERS-DANLOS SYNDROME

• A series of genetic diseases with faulty

assembly of collagens (lysyl hydroxylase
deficiency).

• Hyperextensible skin and hypermobile joints

• In some forms (e.g., type IV), weakness in blood
vessels or intestines are life threatening.
OSTEOGENESIS IMPERFECTA

• Brittle bone disease

• Genetic disorder, at least 9 types (type 1 most
common)
• Weakness of the bones, blue tinge to the whites of
the eye, short height, loose joints, hearing loss,
breathing problems and problems with the teeth,
poor muscle tone
• Underlying cause: mainly lack of collagen I
SCURVY

• Result of lack of vitamin C

• Ascorbic acid is required as a cofactor for prolyl
hydroxylase and lysyl hydroxylase
• Resulting in decreased red blood cells, gum
disease, weak hair, bleeding from the skin, poor
wound healing
• Nowadays rare, usually ocurring in people with
unusual dietary habits, mental disorders,
alcoholism
FIBERS - ELASTIC

• Thin fibers or fenestrated sheets composed of various glycoproteins, including the

protein elastin
• Providing elastic properties to tissues that experience repeated deformation (in skin,
blood vessels, lung, bladder)
• May be stretched up to 150% of their resting length
• Visualized by selectively staining with Weigert’s, resorcin-fuchsin, or aldehyde-
fuchsin stainings
• Electronogram: consist of amorphous core of elastin surrounded by microfibrillar
glycoprotein, fibrillin (8-10nm)
ELASTIN

• Rich in glycine and proline, but contains little or no hydroxyproline and

hydroxylysine
• Uniquely contains desmosine and isodesmosine, which are thought to
cross-link the molecules into a network of randomly coiled chains
• This cross-linking is responsible for its rubber-like properties.
NETWORK OF ELASTIN MOLECULES
MARFAN SYNDROME

• Defect in elastic fibers synthesis (mutation of

the gene coding fibrilin-1)
• Reduced elasticity in skin and lungs
• Skeletal defects (bones are longer and thinner
than usual)
• Cardiovascular complications (aneurism, valve
prolapse)
ELASTIN STAINING

• Elastin staining (“Weigert’s”, “aldehyde

fuchsin”, “Verhoeff”): elastic fibers appear
purple/black
• Collagen fibers stain orange/pink or
blue/green depending on other stains used
(von Gieson’s or trichrome, respectively)
RETICULAR FIBERS

• Reticular fibers provide a supporting framework for the cellular

constituents of various tissues and organs
• Reticular fibers are named for their arrangement in a mesh-like
pattern or network
• Made of type III collagen fibrils (sometimes in association with type IV
collagen)
RETICULAR FIBERS

• Very thin branching fibers (rete—net)

• Diameter = 0.5 to 2 micrometers
• Coating of proteoglycans & glycoproteins– black colour with silver
staining, red with PAS
• Abundant in connective tissue around muscle fibers, nerve fibers,
acini of glands, lymphoid organs & bone marrow
• In most locations, reticular fibres are produced by fibroblasts.
• However, the reticular fibres that support the stroma of hemopoietic
and lymphatic tissue are made by special cells called reticular cells.
RETICULAR FIBERS
CELLS OF CONNECTIVE TISSUE
1. Fibroblasts
Fixed 2. Adipose (fat) cells
(permanent
residents)
3. Tissue Macrophages**
4. Mast cells**

Free
(transient 5. Lymphocytes & Plasma Cells
(differentiated B-cells) **
residents)
6. “Leukocytes”**
(specifically, neutrophils, eosinophils, & basophils)

** derived from hematopoietic stem cells and

involved in immune function and inflammation
CELL LINEAGES
FIBROBLASTS

• Synthesize and secrete components: fibers and ground substance.

• Active and quiescent stages (when quiescent sometimes called fibrocytes or
mature fibroblasts).
• Synthesize growth factors.
• Rarely undergo cell division unless tissue is injured, which activates the
quiescent cells.
• Play a major role in the process of wound healing and respond to an injury
by proliferating and enhanced fiber formation.
FIBROBLASTS – ACTIVE AND MATURE
(FIBROCYTES)
MYOFIBROBLASTS

• Similar to fibroblasts
• Electron microscopy: actin and myosin filaraments, dense bodies
similar to smooth muscle tissue
• External lamina is absent in comparasion to smooth muscle
• Function: wound healing (contraction) and tooth eruption
MAST CELLS

• Principal function is storage in secretory granules and REGULATED

release (degranulation) of histamine and other vasoactive mediators
of inflammation.
• Responsible for the immediate hypersensitivity response
characteristic of allergies, asthma and anaphylactic shock.
MAST CELLS

• Connective tissue mast cells are found in skin (dermis) and peritoneal
cavity; mucosal mast cells are in the mucosa of the digestive and
respiratory tracts.
• Cytoplasm filled with basophilic secretory granules. Similar to
circulating basophils, but recent evidence they actually arise from a
unique progenitor.
MAST CELLS

• Granules contain primary mediators:

• Heparin
• Histamine (or chondroitin sulfates)
• Neutral proteases
• Aryl sulfatase
• Eosinophil and neutrophil chemotactic factor
MAST CELLS

• Mast cells synthetize secondary mediators:

• Leukotrienes
• Thromboxanes
• Prostaglandins
• Platelet-activating factor
• Bradykinins
• Interleukins
• Tumor necrosis factor
MAST CELLS
UNILOCULAR ADIPOCYTES

• Derived from mesenchymal stem cells

• 0.1 mm in diameter (can be twice the size)
• Nucleus is compressed to one side by enormous lipid droplet
• The lipid is extracted by histological processing, and appears as an
empty space
• Energy stored in the form of triglycerides and cholesteryl ester
UNILOCULAR ADIPOCYTES

• Triglyceride storage and glucose metabolism (insulin and glucagon receptors)

• Secretion of many bioactive molecules:
• leptin (regulates satiety)
• adiponectin (regulatingmetabolism of saccharides and lipids, increasing the sensitivity of
tissues to insulin)
• resistin (glucose tolerance and insulin sensitivity)
• angiotensinogen (blood pressure)
• steroids (glucocorticoids & sex hormones)
• growth factors (e.g. insulin-like growth factor, tumor necrosis factor)
• cytokines (e.g. interleukin-6)
UNILOCULAR ADIPOCYTES
UNILOCULAR ADIPOCYTES
BROWN (MULTILOCULAR) ADIPOCYTES

• Many small lipid droplets inside

cytoplasm instead of a single one
• More polygonal in shape
• Nucleus in the center, round, more
mitochondria (cytochrome gives
them the red color)
• Mitochondria of brown fat cells
express uncoupling protein which
“short circuits” the electron transport
chain producing HEAT rather than
ATP.
BROWN ADIPOCYTES
PERICYTES

• Derived from undifferentiated mesenchymal cells

• Surround endothelial cells of capillaries and small venules
• Located outside connective tissue compartment due to their own basal
lamina
• Characteristics of endothelial cells and smooth muscle cells
• May function in contraction
• May differentiate into other cells
MACROPHAGES

• Monocytes escape from blood vessels into connective tissue where they
differentiate into macrophages
• Large phagocytic cells (10 to 30 micrometers) with oval or kidney shaped
nuclei
• Abundant
• Inactive and active (resident and elicited)
• Scavenger cells
• Foreign body giant cells
• Antigen presenting cells
MACROPHAGES

• Different names:
• Kupffer cells – liver
• Dust cells – lung
• Langerhans cells – skin
• Monocytes – blood
• Macrophages - connective tissue
MACROPHAGES
PLASMA CELLS

• Mature B lymphocytes that constitutively secrete antibodies

(humorally mediated immunity)
• Present in chronic inflammation
• Large, ovoid cells, 20 micrometers in diameter, eccentrially placed
nucleus with heterochromatin radiating („clock face”)
• Life span 2-3 weeks
PLASMA CELLS
LEUKOCYTES

• Neutrophils:
• Phagocytose of the bacteria in the inflammatory region
• In comparasion to macrophages: short lived, not presenting antigens
• Formation of pus
• Eosynophils:
• Attracted to inflammatory and allergic reaction
• Combat parasites by releasing cytokins
• Phagocytose antibody-antigen complex
LEUKOCYTES

• Basophils:
• Similar to mast cells
• Release of agents that initiate, maintain and control inflammatory proces
• Lymphocytes:
• Present very rarely
• Common in chronic inflammation
NEUTROPHILES
TYPES OF EMBRYOTIC CT

• Mesenchymal connective tissue:

• Present in embro
• Mesenchymal cells in amorphous ground substance containing scattered
reticular fibers
• Frequent mitotic figures – gives rise to mose of loose conecting tissue
• In adult – pulp of teeth, pericytes – can differeciate into other cells
TYPES OF EMBRYOTIC CT

• Mucous tissue:
• Amorphic connective tissue
• Jelly-like matrix composed of hyaluronic acid and t. I and III collagen fibers and
fibroblasts
• Wharton’s jelly
• Umbilical cord, subdermal tissue of embryo
EMBRYOTIC CT
TYPES OF CT PROPER

• Loose (areolar) connective tissue:

• delicate, vascularized, cellular
• supports the epithelia of the major organs and glands
• fills the space between muscle tissue
• Loose CT of mucous membranes – lamina propria
• facilitates transport of cells and materials (secretion, absorption, immunity)
• not very resistant to stress
LOOSE (AREOLAR) CONNECTIVE TISSUE
TYPES OF CT PROPER

• Dense CT:
• Many more fibers than cells
• Dense irregular:
• Meshwork of coarse fibers
• Fibroblasts located interstices between collagen bundles
• Dermis of skin, sheths of nerves, capsules of the spleen, testes, ovary, kidney,
lymph nodes
• Resists multi-directional forces
DENSE IRREGULAR CT
Collagen

Fibroblast nucleus
TYPES OF CT PROPER

• Dense regular:
• collagenous: fibers aligned in defined pattern;
• tendons, ligaments, etc.
• resists linear mechanical stresses
• Dense elastic:
• Branching elastin and microfibrils (fibrillin) with few collagen fibers forming fenesterated
membranes (paralel to one another)
• Elasticity
• In large blood vessels, ligamenta flava of the vertebral column, suspensory ligament of the
penis
DENSE REGULAR CT
DENSE ELASTIC CT
WHITE ADIPOSE TISSUE

• Distributed throughout the body, especially in hypodermis of skin,

underlying abdomen, buttocks, axilla, thigh, breast.
• Age and sex determine density and distribution of subcutaneous layer
of adipose tissue.
• Internally, preferentially located in mesentary and retroperitoneal
space, around kidneys and in bone marrow.
• Highly vascularised
WHITE ADIPOSE TISSUE

• Functions:
• Energy storage
• Insulation from heat loss
• Protection of internal organs from physical trauma
• Producing hormones
BROWN (MULTILOCULAR) ADIPOSE TISSUE

• Present mostly in newborns (and hibernating

mammals) and involved in thermoregulation
• Present also in adults, amount decreases with
time (present mostly in the neck)
• More reddish due to cytochrome and a lot of
blood vessels
• „Classical”:between the shoulder blades,
armpits, surrounding the kidneys, the neck, and
supraclavicular area, and along the spinal cord
BROWN (MULTILOCULAR) ADIPOSE TISSUE

• „Dispersed”: distributed throughout white

adipose tissue
• Cold exposure promoting development and
activation
CARTILAGE

• Strong and some pliable tissue

• Avascular,
• No lymphatic vessels
• Not innervated
• Chondrocytes - they are large cells ofrounded shape with a central
spherical nucleus, the cytoplasm is basophilic and rich in rough
endoplasmic reticulum, glycogen, lipid droplets
CARTILAGE

• Cells are housed in small spaces within the matrix known as lacunae
• Fibers can be solely collagenous or a combination of collagenous and
elastic depending upon the cartilage type
• The surface of the hyaline and elastic cartilage is enveloped by a
connective tissue membrane - the perichondrium, from which the
growth and nutrition of cartilage is provided
• The fibrocartilage has no perichondrium
CLASSIFICATION OF THE CARTILAGE

• hyaline cartilage
• elastic cartilage
• fibrocartilage
HYALINE CARTILAGE

• Firm and bluish - gray

• In the microscope, its matrix appears clear (glasslike) because fibers and
ground substance have the same staining capacity and refractive index
• In the adult, chondrocytes are arranged in groups, composed of 2, 4, or 8
cells that are of the same origin (cells of each group have been derived
from the same parent cell) such cell groups are termed isogenous groups or
cell nests
• Contains type II collagen, perichondrium present (mostly)
• Is located in: larynx, nose, ventral ends of ribs (sternum), tracheal rings,
bronchi, articulating surfaces of the joints of the body, also – epiphyseal
plates of growing bones
HYALINE CARTILAGE

• Fibrous component is represented by very fine collagenous fibers that form

a feltwork
• May be visualized by tryptic digestion or polarized light microscopy
• An amorphous ground substance appears homogeneous and is composed
of chondromucoid
• The ground substance immediately surrounding each group of isogenous
cells usually stains more deeply than elsewhere:
• several isogeneous groups may unite in complexes termed as territoria
• between territoria the matrix stains slightly and is called as interterritorial matrix
HYALINE CARTILAGE
ELASTIC CARTILAGE

• Usually yellowish in colour and opaque and more flexible than hyaline
• Chondrocytes are numerous and scattered singly or in small isogenous groups of
two or three cells that are surrounded by capsules of the intercellular substance
• Fibers: elastic and collagenous, the matrix (intercellular substance) shows
characteristic fibrillar appearance in H.E. preparations
• This is caused by presence of elastic fibers that are never totally masked by
amorphous ground substance
• Elastic fibers may be visualized by special dyes as orcein or resorcin
ELASTIC CARTILAGE

• Located in the pinna of the ear, external and internal auditory tubes,
epiglottis, larynx
• Contains type II collagen
• Perichondrium is present
ELASTIC CARTILLAGE ORCEIN
FIBROCARTILAGE

• Transitional tissue between hyaline cartilage and dense fibrous

connective tissue of tendons and ligaments
• The intercellular substance is composed of thick bundles of
collagenous fibers running parallel with one another
• Chondrocytes are not numerous, rounded and are arranged singly or
in pairs in rows between bundles of collagenous fibers
FIBROCARTILAGE

• Amorphous ground substance is inconspicuous and found only in the

immediate vicinity of the chondrocytes
• Fibrocartilage has no true perichondrium
• Present in intervertebral disks, pubic symphysis and attachment to the
bone
• Contains type I colagen
FIBROCARTILAGE