HISTOLOGY // CONNECTIVE TISSUE PROPER

HISTOLOGY Trans No.: 1.3

CONNECTIVE TISSUE PROPER
Lecturer: DR. TOLENTINO Date: June 28, 2013
Transcribed by: Gonzaga, Paruñgao & Solis
Fibroblasts / Fibrocytes
CONNECTIVE TISSUE (CT)
 Abundance of extracellular matrix (ECM) and a paucity of  Most common cell in the CT proper
cells  Functions:
 Most are derived from the mesenchyme of the mesoderm 1. synthesis of proteins in the ground substance
 2 major groups: 2. target of growth and differentiation factors
1. Connective tissue proper 3. wound healing (mitosis is rare in adults, but can
2. Special types of connective tissue resume for this purpose)

 Functions: Table 1. Histological differences between active fibroblasts and

1. Envelops muscles inactive fibrocytes
2. Binds body parts together while allowing for some FIBROBLAST FIBROCYTE
degree of movement of these parts in relation to (active) (quiescent/idle)
their immediate anatomical neighbor Size Large Small
3. Forms the stroma/ supporting framework of various Shape More elongated Spindle-shaped
organs Cytoplasmic Many Few
4. Serves as a venue for the exchange of nutrients processes
and waste products between the cells and the
Nucleus Large, ovoid, Small, elongated,
blood supply (via diffusion)
pale-staining dark
5. Serves as a reservoir for cell growth and
Mitochondria Rich Poor
differentiation factors
, rER & Golgi
 Components (Outline):
I. Cells
A. Resident/ Fixed (FARMM) Adipose Cells / Adipocytes*
1. Fibroblast & Fibrocyte
2. Adipose cells  Start as lipoblasts derived from mesenchyme
3. Reticular Cells  Spherical/ polyhedral with signet ring appearance
4. Mast cells  Mature adipocytes do not divide; only the volume of fat
5. Macrophages droplets increase
B. Visiting/ Wandering (PLM)  Wide space in light microscopy (LM): represents lipid
1. Plasma Cells droplet lost in staining/ slide preparation
2. Leukocytes  Functions:
3. Macrophages 1. fat storage
II. ECM 2. production of heat
A. Fibers * discussed more in AdiposeTissue section
1. Collagen Fibers
2. Elastic Fibers Reticular Cells
3. Reticular Fibers
B. Ground Substance
 Stellate, with long cytoplasmic processes
 Large, central, lightly staining nucleus with prominent
CELLS OF CT
nucleolus
 Functions:
Mesenchymal Cells 1. Production of reticular fiber network in
hematopoietic tissues and parenchyma of certain
 Highly undifferentiated (pluripotent), migratory embryonic organs (e.g. lymphoid tissue & liver)
cells 2. phagocytosis of antigens and cellular debris
 Precursor of most CT cells
 Elongated, stellate cells with many thin cytoplasmic Mast Cells
processes & an oval euchromatic nucleus
 Found in the lamina propria of the gastro-intestinal tract,
respiratory tract, skin, and small blood vessels

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HISTOLOGY // CONNECTIVE TISSUE PROPER

 LM: large, oval or spherical cells with fine, regular, Table 2. Macrophage-like members of the mononuclear
basophilic granules; distinguished by special stain phagocyte system found in various organs
(toluidine blue) NAME OF CELLS LOCATION
o Metachromasia: ability of granules (acidic) to Monocytes Blood
change the color of basic dyes from blue  Histiocytes Connective tissue
purple/red
Fixed Lymphoid organs, lungs,
 Electron microscopy (EM): granules are electron-dense
macrophages serous cavities
and obscure the nucleus
Multinuclear giant Connective tissue (fusion of
 Function: triggers the allergic response (immediate
cells several macrophages)
hypersensitivity reactions) by degranulation and release
of bioactive substances: Dust cells/ Alveolar Alveoli of lungs
o Heparin: locally acts as a weak coagulant macrophages
o Histamine: potent inflammatory mediator; dilates Kupffer cells Liver
blood vessels and increases their permeabiliy  Microglia Neural tissue (CNS)
local edema & migration of WBCs Epitheloid cells Granulomas
o Serine proteases*: activate several inflammatory Osteoclasts Bone
mediators Sinusoidal lining Spleen
o Chemotactic factors*: eosinophil & neutrophil cells
attractants Langerhans cells Epidermis
o Leukotrienes C4, D4 & E4* (or the slow-reacting
Dendritic cells Lymph nodes
substance of anaphylaxis, SRS-A): triggers smooth
muscle contraction
* - not part of Dr. Tolentino’s lecture; from Mescher
(2010) Plasma Cells
 MAST CELLS vs. BASOPHILS: differ in progenitor cells;
basophils are smaller and not capable of division; mast  Derived from antigen-stimulated B lymphocytes
cells are bigger and mitotic  Function: principal producers of circulating antibodies
(immunoglobulins)
Macrophages  LM: large, ovoid cells with basophilic cytoplasm and
eccentric nucleus with characteristic “clock-face”  due
 Large, stellate cells with an eccentric (peripherally located) to heterochromatin clumps alternating with lighter
oval or kidney-shaped nucleus; may resemble fibroblasts euchromatic ones apposing the nuclear envelope
when inactive  EM: clear juxtanuclear area (“cytocenter”) with well-
 Derived from monocytes; from the blood, these cells developed Golgi complex, rER,and centrioles
migrate into connective tissues to differentiate into
macrophages Leukocytes (White Blood Cells)
 Functions:
1. phagocytose cell debris, abnormal ECM elements,  Migrate to connective tissue from bone marrow via blood
and foreign organisms vessels; they pass in between the endothelial cells that line
2. serve as antigen-presenting cells capillaries and venules in a process called diapedesis 
3. participate in cell-mediated resistance to infections esp. during inflammation
(bacterial, viral, fungal, protozoan, metazoan {e.g.  Includes lymphocytes, monocytes, neutrophils, eosinophils,
parasitic worms}) and tumors and basophils
4. also participate in extrahepatic bile production, iron  Do not return to blood after connective tissue migration
and fat metabolism, and destruction of old EXCEPT for lymphocytes that circulate the blood, lymph,
erythrocytes lymphatic organs, and the interstitial fluid of connective
 LM: may be distinguished by materials they phagocytosed tissue
(e.g. dye)  Function: defense against bacterial invasion or foreign
 EM: irregular surface characterized by pleats, protrusions, matter
and indentations (due to their pinocytotic/ phagocytic
activities); with well developed Golgi apparatus & rER and ECM: FIBERS OF CT
many lysosomes
 Have inactive and active forms
Collagen Fibers
 May be fixed (resident) & immunologically inactive OR free
(wandering, visiting) & inflammatory
 Mononuclear phagocyte system: family of monocyte-  Collagen: most abundant protein in the human body,
derived cells, including the macrophages of the connective accounts to 30% of its dry weight
tissue (a.k.a histiocytes)  Collagen fibers: most abundant fibers found in almost all
connective tissues of all organs
 Tough, thick fibrous proteins that are non-branching and
often collect to form fiber bundles (0.5-15 µm diameter)

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HISTOLOGY // CONNECTIVE TISSUE PROPER

 With tensile strength comparable to steel; flexible but  Have lesser tensile strength than collagen fibers; very
relatively inelastic supple
 Colorless in freshly prepared tissue  Unstained in fresh tissue preparations, but become
 LM: acidophilic/ pinkish (H&E technique/ hematoxylin & yellowish if abundant
eosin dye technique); green (Masson’s trichome technique);  Stain blue to black in Orcein staining procedure
type III collagen fibers may also be viewed using silver  Form sparse networks interspersed with collagen bundles
staining in organs subject to bending or stretching (e.g. wall of large
 EM: fibers and fibrils have stripes at 64 nm-intervals along arteries)
their lengths that reflect the staggering of tropocollagen  Found in the ligamenta flava, extracellular spaces of the
molecules that make them up elastic cartilage, interalveolar septa, bronchi and
 4* most frequently recognized fibers in histologic slides bronchioles, vocal ligaments, and the dermis
(from Eroschenko, 2013):
o Type I collagen fibers Table 3. Summary of differences between collagen fibers,
 Most common; very strong and offer great reticular fibers, and elastic fibers
resistance to tensile stresses FEATURE COLLAGEN RETICULA ELASTIC
 Found in the dermis, in tendons, ligaments, FIBER R FIBER FIBER
fascia, capsules of organs, fibrocartilage, and
Abundanc +++ + ++
bones
e
o Type II collagen fibers
 Provide resistance to pressure Thickness 0.5-15 µm 0.5-2 µm 0.1-10 µm
 Found in hyaline cartilage, elastic cartilage, LM ~straight, delicate branched,
and in the vitreous of the eye Appearanc arranged in network; netlike; blue
o Type III collagen fibers e bundle of black in to black in
 Thin, branching reticular fibers that form a fibrils, silver orcein
delicate supporting meshwork in some organs eosinophilic; impregnatio
 Found in lymph nodes, spleen, and bone green in n
marrow Masson’s (argyrophilic
o Type IV collagen fibers trichome )
 Occur as meshwork in the basal lamina, to Mechanica high tensile pliable pliable &
which basal portions of cells attach via l Property strength; elastic
hemidesmosomes tough
* - there are more than 4 types Molecular Tropocollag collagen microfibrils
Structure en type III; & elastin;
Reticular Fibers molecules heavily isodesmosi
 collagen glycosylated ne and
 Made of collagen type III fibrils  desomosine
 Very thin (0.5-2 µm diameter) branching and anastomosing collagen crosslink
fibers forming extensive networks around parenchymal fibers elastin
cells of various organs (e.g. liver & endocrine glands) chains
 Particularly abundant in hematopoietic organs (e.g. spleen, Location different liver, dermis,
lymph nodes & red bone marrow stroma); produced by types, hematopoiet ligamenta
fibroblast-like cells called reticular cells different ic organs, flava, etc.
 Loose disposition creates a flexible network in organs locations sinusoids,
subject to changes in form or volume (e.g. arteries, uterus liver
& intestinal muscle layers)
 Usually not visible in H&E preparations
 Stained black by impregnation with silver salts, making ECM: GROUND SUBSTANCE
them argyrophilic  Highly hydrated, transparent, complex mixture of
 Heavily glycosylated, making them periodic acid-Schiff macromolecules
(PAS)-positive  Principally in 3 classes: glycosaminoglycans,
proteoglycans, and multiadhesive glycoproteins
Elastic Fibers  Fills in spaces between cells and fibers of connective tissue
 Elastin + numerous microfibrils branching and  Function: acts as both lubricant and a barrier to the
anastomosing network of elastic fibers in random coil penetration of invaders
conformation
 Fibroblasts usually synthesize elastic fibers, but in walls of Glycosaminoglycans (GAGs)
large vessels, they are synthesized by the smooth muscle  Formerly called mucopolysaccharides
cells  Linear polysaccharides formed by repeating disaccharide
 Adapt a linear conformation when stretched, but recoil units usually composed of a uronic acid and a hexosamine
when released  Hexosamines can either be: glucosamine or
galactosamine
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HISTOLOGY // CONNECTIVE TISSUE PROPER

 Uronic acid can be: glucoronic or iduronic acid CLASSIFICATION OF THE CONNECTIVE TISSUE PROPER
 Hyaluronic acid (hyaluronan): largest, most, unique, most
ubiquitous GAG  Composed of the cells, fibers, and ground substance
o Long polymer of the disaccharide glucosamine- components
glucuronate
 Four main GAGs found in proteoglycans:
1. Dermatan sulfate
2. Chondroitin sulfate
3. Keratan sulfate
4. Heparan sulfate

Proteoglycans

 Composed of a core protein to which are covalently

attached various numbers and combinations of the sulfated
GAGs
 Synthesized in the rER, mature in the Golgi, and secreted
from cells by exocytosis
 In cartilage, the core proteins of secreted proteoglycans are
bound via small link proteins to a hyaluronic acid chain,
forming much larger structures – proteoglycan
aggregates
 Aggrecan: one of the most important proteoglycans found
in the ECM and the most dominant proteoglycan in
cartilage.
o Core protein bears several chondroitin sulfate and
keratan sulfate chains and is in turn bound via a link
protein to hyaluronic acid
 Syndecan: cell surface proteoglycans present on many
types of cells, particularly epithelial cells
Multiadhesive glycoproteins
 Have carbohydrates attached but in contrast to
proteoglycans the protein usually predominates
 Fibronectin: synthesized by fibroblasts and some epithelial
cells
 Have binding sites for collagens, certain GAGs, and
integrins of cell membranes
 Interactions at these sites help to mediate normal cell
adhesion and migration and cause fibronectin to be
distributed as a network in the intercellular spaces of many
tissues
 Laminin: larger, trimeric, cross-shaped glycoprotein that
participates in the adhesion of epithelial cells to the basal
lamina
 Cells interact with ECM components by using cell-surface
molecules called matrix receptors that bind to collagen,
fibronectin and laminin
 Integrins: family of transmembrane linker proteins
o Bind their ligands in the ECM with relatively low
affinity, allowing cells to explore their environment
without losing attachment to it or becoming glued to
it
o Interact with actin microfilaments (mediated by talin
and vinculin)
Loose (Areolar) Connective Tissue
 Supports structures which are normally under some
pressure and low friction
 Usually supports epithelial tissue
 Forms a layer around small blood and lymphatic vessels
 Fills spaces between muscle and nerve fibers
 Has all the main component s of connective tissues in
roughly equal parts
 Most numerous cells are fibroblasts and macrophages
 Collagen, elastic, and reticular fibers are present
 Has delicate consistency, flexible, well vascularized, and
not very resistant to sress
 Found in the papillary layer of the dermis, in the
hypodermis, in the linings of the peritoneal and pleural
cavities, in glands, and in the mucous membranes
supporting epithelial cells
Dense Connective Tissue
 Adapted to offer resistance and protection
 Fewer cells and a clear prominence of collagen fibers over
ground substance
 Less flexible and far more resistant to stress
 Dense irregular connective tissue: When the collagen
fibers are arranged in bundles without a definite
orientation
o Collagen fibers form a 3D network of dense
irregular tissue, providing resistance to stress in
all directions
o Found closely associated with loose connective
tissues
 Dense regular connective tissue: arranged according to
a definite pattern.

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HISTOLOGY // CONNECTIVE TISSUE PROPER
o This arrangement offers great resistance to
traction forces
o Found in tendons and ligaments
o Fibrocytes contain elongated nuclei parallel to the
fibers
o Poorly vascularized (repair of tendons is very
slow)
o Collagen bundles of tendons vary in size and are
enveloped by small amounts of loose connective
tissue containing small blood vessels and nerves
o The tendon is surrounded by a sheath of dense
irregular connective tissue (sheath made up of
two layers, boith lined by synovial cells)
Modified Loose Connective Tissue
Reticular Tissue
 Consists of reticular fibers of Type III collagen that are
produced by reticular cells
 The heavily glycosylated reticular fibers provide the
architectural framework that creates special
microenvironments for hematpoietic organs and lymphoid
organs (bone marrow, lymph nodes, and spleen)
 Reticular cells are dispersed along this framework and
partially cover the reticular fibers and ground substance
with cytoplasmic processes
 Cells of the mononuclear phagocyte system are
strategically dispersed along the trabeculae
 Monitor the slow flow of materials through the sinus-like
spaces and remove invaders by phagocytosis
Mucous Tissue
 Found mainly in the umbilical cord and fetal tissues
 Has the abundance of ground substance composed
mainly of hyaluronic acid (making it jellylike with very few
collagen fibers and scattered fibroblasts)
 Wharton’s Jelly: mucous tissue found in the umbilical
cord
 Also found in the pulp cavity of young teeth
Adipose Tissue
 Specialized type of connective tissue in which
adipocytes or fat cells predominate
 Can be found in isolated or in groups within loose or
irregular connective tissues
 Represents 15-20% of the body weight in men of normal
weight; in women of normal weight, 20-25% of body
weight
 The most widely studied cell of connective tissue
(because of the growing worldwide epidemic of obesity
and its associated problems, including diabetes and heart
disease)
 Triglycerides: the neutral fats
 It is in a state of continuous turnover and is sensitive to
both nervous and hormonal stimuli
 Functions:
1. Key regulators of the body’s energy metabolism;
largest repository of energy
2. Release hormones and a number of important
factors
3. Recognized as a major endocrine and signaling
organ
4. Poor heat conductor so it contributes to the
thermal insulation of the body
5. Helps keep organs in place
6. Helps shape the surface of the body
7. Act as shock absorbers in the palms and soles of
the feet
 2 types: white adipose tissue and brown adipose
tissue
I. White Adipose Tissue
 Specialized for long-term energy storage
 Spherical (when isolated) or polyhedral (when closely
packed in adipose tissue) cells with peripheral nuclei
 Classified as unilocular because triglycerides are stored
in a single locus; large, lipid droplet makes up 85% of the
cell’s weight
 Signet ring: how a unilocular adipose cells appears in
standard microscope preparations
 The thickest portion of the cytoplasm surrounds the
nucleus of these cells and contains a Golgi apparatus,
mitochondria, poorly developed cisternae of the rER and
free polyribosomes
 Each adipose cell is surrounded by a thin external basal
lamina
 Color of freshly dissected white adipose tissue depends
on the diet and varies from white to bright yellow, due to
the presence of carotenoids dissolved in the fat droplets
 Age and gender determine the distribution and density of
adipose deposits
 In the newborn, white adipose tissue has a more uniform
thickness throughout the body
 Distribution is regulated by sex hormones, which control
adipose deposition in the breasts and thighs
II. Brown Adipose tissue
 Color is due to the numerous mitochondria scattered
through the adipocytes and the large number of blood
capillaries in this tissue
 Multilocular, because they contain many small lipid
inclusions
 The many small lipid droplets, abundant mitochondria,
and rich vasculature all help mediate this tissue’s
principal function of heat production
 Has much more limited distribution compared to white
adipose tissue
 Cells of brown adipose tissue are polygonal and
generally smaller than white adipose cells but their
cytoplasm contains a great number of lipid droplets of
various sizes
 Nuclei are spherical and are centrally located
 Numerous mitochondria have abundant long cisternae
 Functions:
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1. Produce heat by nonshivering thermogenesis

2. Found in hibernating mammals (once called
the hibernating gland)
3. Also found in newborn mammals
 Thermogenesis in brown adipose:
1. Nerve impulses liberate norepinephrineinto brown
adipose tissue
2. Activation of hormone-sensitive lipase present in
adipose cells, promoting hydrolysis of triglycerides
to fatty acids and glycerol
3. Liberated fatty acids are quickly metabolized, with a
consequent increase in oxygen consumption and
heat production
4. Elevated temperature of the tissue and warming the
blood passing through it
o Heat production is increased in these cells
because the mitochondria have in their inner
membrane a transmembrane protein calle
thermogenin or uncoupling protein (UCP-1),
a marker unique to brown fat
o Warmed blood circulates throughout the body,
distributing the heat and carrying fatty acids not
metabolized in the adipose tissue for use
elsewhere

Elastic Tissue
 Arranged in a parallel manner
 Laid down by fibroblasts
 Found in the ligament flava and suspensory ligament of
the penis

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