Connective Tissues

Dr Nyango P. B
Dept of Human Anatomy
Faculty Of Medical Sciences
University of Jos.
Learning Objectives
1. Know and Understand Connective Tissues.
2. Classify Connective Tissues.
3. Know the Functions and Properties of
Connective tissues.
4. Types of Connective Tissue Cells
5. Ground Substance Composition
6. Connective Tissue Staining
7. Clinical Significance of Studying Connective
Tissues.
Lecture Outline
• Introduction
• Notable Scientists
• Functions of Connective Tissue
• Composition of Connective Tissue
• Classification of Connective Tissue
• Clinical Relevance
INTRODUCTION
• Connective tissue fills the spaces between organs and tissues, and
provides structural and metabolic support for other tissues and organs.
• Connective tissue is made up of cells and extracellular matrix. The
extracellular matrix is made up of fibres in a protein and
polysaccharide matrix, secreted and organised by cells in the
extracellular matrix. Variations in the composition of the extracellular
matrix, determines the properties of the connective tissue. For
example, if the matrix is calcified, it can form bone or teeth.
Specialised forms of extracellular matrix also makes up tendons,
cartilage, and the cornea of the eye. General connective tissue is
either loose, or dense, depending on the arrangment of the fibres. The
cells sit in a matrix made up of glycoprotein's, fibrous proteins and
glycosoaminoglycans, which have been secreted by the fibroblasts, and
the major component of the matrix, is in fact, water.
NOTABLE SCIENTISTS
Ira Thompson Van Gieson (1866
Richard von Volkmann (1830–1889) – 1913 )
• Richard von Volkmann (1830– • was an American
1889), one of the most neurologist, psychiatrist,
important surgeons of the 19th bacteriologist and
century, is regarded as one of
neuropathologist.
the fathers of orthopaedic
surgery. • The red staining method of
• Volkmann’s syndrome I (or collagen
Volkmann’s disease)Congenital • Van Geison Method
talus luxation. Also referred to
as tibiotarsal dislocation
causing a congenital deformity
of the foot.
FUNCTIONS OF CONNECTIVE TISSUE
• Holds together structures like skin, muscles,
blood vessels e.t.c
• Facilitates movement between structures
• Provides support to the brain and spinal cord
• A store of nutrition
• Immune system service
• Inflammation and Wound repair service
CONNECTIVE TISSUE COMPOSITION
• Cells • Cells
• Matrix – Intrinsic Component
– Fibers Cells
• Fibroblasts
– Intercellular Ground
• Undifferentiated
Substance Mesenchymal Cells
• Fibers – Migrant Related
– Collagen Fibers • Macrophage Cells
– Reticular Fibers • Mast Cells
– Elastic Fibers • Granulocytes
• Agranuloctes
 Fibers in Connective Tissue
• Collagen
– most abundant protein in human body (up to 30% dry
weight)
– multiple types: fibril-forming or fibril-associated (in skin,
tendon, cartilage, bone, dentin, blood vessels); cross-
linked networks (in all basement membranes)
• Reticular Fibers – specialized type of collagen (Type III;
reticulin) associated with smooth muscle in organs subjected
to changes in volume, forms the stroma in lymphatic and
hematopoietic organs
• Elastic Fibers –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)
 Major Collagen Fiber Types (out of at least 20)

Collagen Type Tissues Function

Fibril-forming collagens (these are visible)
I
Skin, tendon, bone, dentin Resistance to tension
(most abundant)

II Cartilage, vitreous of eye Resistance to pressure

III Skin, muscle, blood vessels, Structural framework and

(reticulin) liver, etc. stability

Network-forming collagens

IV All basement membranes Support and filtration

Fibril-associated collagens with interrupted triple helices (FACIT)

VI, IX Assoc. w/ type I and II fibrils Fibril-fibril / fibril-ECM binding

Anchoring filament collagens

VII Epithelia Epidermis to basal lamina

Source Undetermined
 Collagen fibers viewed by light microscopy
H&E Trichrome

University of Michigan Histology Collection University of Michigan Histology Collection

 Assembly of collagen fiber bundles

Junquiera and Carneiro. Basic Histology. Tenth Edition. 2003. Figure 5-19.
 Basement Membrane –
Collagen Types IV, VII, and III
• Basement membranes are sheets of extracellular
matrix proteins located at the interface of
parenchyma (epithelia, endothelia, muscle, nerves,
adipocytes) and connective tissue / ECM.
• Main constituents are glycosaminoglycans (heparan
sulfate), fibrous proteins (collagen types IV, VII, III),
structural glycoproteins fibronectin, laminin and
entactin.
• This is NOT a plasma membrane.
 Basement membranes vary in thickness
Thick Thin -- requires special stain to visualize

BM

BM
BM
University of Michigan Ross,M, Pawlina, W. Wheater’s Functional Histology:
Histology Collection A Text and Atlas. Fifth Edition. 2006. Figure 4.4.

Intestinal glands, PAS

trachea, H&E PAS reacts with carbohydrate-rich molecules such as perlecan, laminin
and type III collagen associated with the basement membrane.
 Basement Membrane(LM): Three layers in the EM

hemidesmosomes

basal lamina
1. lamina lucida
(LL) or rara 10-50
nm
2. lamina densa
(LD) 20-300 nm
(type IV collagen)

fibroreticular lamina
3. Fibroreticular
FL LL LD lamina (FL)
merges with
underlying CT
(type III* and type VII
collagen fibrils)
Connective tissue
*basement membranes can also
Source Undetermined be visualized with silver stain

So, the “basement membrane” is the basal lamina + the fibroreticular lamina
 Noncollagen Components of the
Extracellular Matrix

• Elastin
• “Ground substance”
– Glycosaminoglycans (GAG’s)
– Proteoglycans
– Multiadhesive matrix proteins
• laminin
• fibronectin
Elastic Fibers
LM: Visualized by selectively staining with Weigert’s, resorcin-fuchsin, or
aldehyde-fuchsin

EM: Consist of amorphous core of elastin surrounded by microfibrillar

glycoprotein, fibrillin (8-10nm).

Elastin: is rich in glycine and proline, but it 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.

Confers elasticity: present in large amounts in ligaments, lung, skin,

bladder, and walls of blood vessels.
Network of elastin molecules can stretch
and recoil like a rubber band

Alberts et. al. Molecular Biology of the Cell.

 Elastin appears amorphous (not fibrillar) in
the electron microscope

Source Undetermined

Ross,M, Pawlina, W. Wheater’s Functional Histology:

Image of elastin
A Text and Atlas. Fifth Edition. 2006. Figure 4.4. removed
E=elastin C, collagen fibrils
M/L=microfibrils of fibrillin, a scaffolding
glycoprotein involved elastin deposition
Marfan Syndrome: defect in fibrillin gene, results in
weakened elastic fibers
 Elastic and Collagen Fibers

University of Michigan Histology Collection University of Michigan Histology Collection

elastin stain (“Weigert’s”, “aldehyde fuchsin”, “Verhoeff”):

H&E stain: collagen stains orange/pink; elastic fibers stain elastic fibers are purple/black
glassy red (generally only visible if in HIGH abundance) collagen fibers stain orange/pink or blue/green depending on
other stains used (von Gieson’s or trichrome, respectively)
Reticular Fibers
• A variety of collagen fibers type III
• Much finer with uneven thickness
• Stained by silver impregnation black.Collagen I
stains brown.
• Provides support in lymphoid organs
• An Essential part of all basement membranes
• Related to smooth muscles and nerve fibres
• Synthesized by Fibroblasts and Reticular cells
 Cells in Connective Tissue

1. Fibroblasts
Intrinsic 2. Undifferentiated Mesenchymal
Component 3. Adipose (fat) cells

Migrant 4. Lymphocytes & Plasma Cells

(differentiated B-cells) **
Related 5. “Leukocytes”**
6. Tissue Macrophages
7. Mast Cells
(specifically, neutrophils, eosinophils, & basophils)

** derived from hematopoietic stem cells and

involved in immune function and inflammation
 Connective Cell Lineages

Junqueira and Carneiro. Basic Histology. Tenth Edition. 2003.

 Fibroblasts are the most common cells in connective
tissue
• Synthesize and secrete components of the ECM:
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.
Image of active and
inactive fibroblasts
Active and inactive
removed
fibroblasts

(Both images) Ross, M. Pawlina, W. Wheater’s Functional Histology: A Text and Atlas. Fifth Edition. 2006.
 Adipocytes predominate in adipose tissue
Very active cells with many functions:
• Triglyceride storage and glucose metabolism (insulin and glucagon receptors)
• Secretion of many bioactive molecules:
leptin (regulates satiety) angiotensinogen (blood pressure) steroids (glucocorticoids & sex hormones)
growth factors (e.g. insulin-like growth factor, tumor necrosis factor ) cytokines (e.g. interleukin-6)

Single, large
lipid droplet

Ross, M. Pawlina, W. Wheater’s Functional Histology: A Text and Atlas. Fifth Edition. 2006.

White (common, yellow, unilocular) adipose tissue stained with Masson’s trichrome
 Adipocytes

Lipid (fat)
droplet

Nucleus

Capillaries
Source Undetermined
 Monocytes escape from blood vessels into connective
tissue where they differentiate into Macrophages

University of Michigan Histology Collection

Primary function: phagocytosis and antigen presentation

 Plasma Cells are mature B lymphocytes
that constitutively secrete antibodies

University of Michigan Histology Collection Junqueira and Carneiro. Basic Histology.

Tenth Edition. 2003. Figure 6.5.

Black arrows indicate several plasma cells White arrows = Golgi regions
 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.
• Connective tissue mast cells
are found in skin (dermis) and
peritoneal cavity; mucosal
Junqueira and Carneiro. Basic Histology. Tenth Edition. 2003.
Figure 5.10.
mast cells are in the mucosa of
the digestive and respiratory
Metachromasia – when stained with
tracts.
toluidine blue, the granules bind the
dye and change its color to red.
 EM of a Mast Cell

Junqueira and Carneiro. Basic Histology. Tenth Edition. 2003. Figure 5.11.
 Mast Cell Secretion

Junqueira and Carneiro. Basic Histology. Tenth Edition. 2003. Figure 5.12.
•
Neutrophils
Enter connective tissue from blood vessels as the “first wave” in acute inflammatory responses
• Small cells with multi-lobed, heterochromatic nuclei (aka “polymorphonuclear neutrophils”, “PMNs”, “polys”)
• Primary function: anti-bacterial (are phagocytic like mphages, but SHORT-lived and NOT antigen presenting)

University of Michigan Histology Collection

 Ground Substance of the
Extracellular Matrix (ECM)
1. Glycosaminoglycans (GAG)
• linear (unbranched) polysaccharides, e.g. heparan sulfate, condroitin sulfate,
keratan sulfate, hyaluronic acid
• very hydrophilic due to abundant negative charges (e.g. SO4- groups).
• except for hyaluronic acid, are usually bound covalently to protein core as part
of a proteoglycan

2. Proteoglycans
• core protein + GAG side chains (like a bottle brush)
• bind cells, other proteins, and/or ECM components

3. Multiadhesive glycoproteins
• small glycosylated proteins containing NUMEROUS binding sites to cells,
signaling molecules, and other ECM components
• e.g. fibronectin and laminin: important for adhesion of epithelial cells to the
basal lamina via transmembrane integrin receptors.
CLASSIFICATION
• EMBRYONIC CONNECTIVE TISSUES
• Mesenchyme
• CONNECTIVE TISSUE PROPER
• Loose irregular connective tissue
• Dense irregular connective tissue
• SPECIALISED CONNECTIVE TISSUES
• Dense regular connective tissue which is found in
tendons and ligaments.
• Cartilage
• Adipose Tissue
• Haemopoietic tissue (bone marrow, lymphoid tissue)
• Blood
• Bone
Loose Connective Tissue
Includes;
1. Areolar tissue,
2. Reticular Tissue and
3. Adipose Tissue

• Loose connective tissue is named based on the weave and type of its
constituent fibers. There are three main types of connective tissue fiber:

– Collagenous : Collagenous fibers are made of collagen and consist of bundles of fibrils
that are coils of collagen molecules.

– Elastic : Elastic fibers are made of elastin and are "stretchable."

– Reticular : Reticular fibers consist of one or more types of very thin collagen fibers.
They join connective tissues to other tissues.
 Loose connective tissue:
delicate, vascularized, flexible; facilitates transport of cells and materials
(secretion, absorption, immunity)

p r ia
a pro
i n
lam

University of Michigan Histology Collection University of Michigan Histology Collection

small intestine lamina propria mammary gland intralobular connective tissue

Loose Connective Tissue
• Adipose Tissue-Aggregation of fat cells,also
called adipocytes
• Distribution
– Panniculus adiposus(subcute fat layer)
– Hollow spaces in the body(orbit,marrow)
– Around abdominal Viscera
Dense Connective Tissue
Includes;
1.Tendons and
2.Ligaments.
• Also called dense fibrous tissue, a type of with
fibers as its main matrix element. The fibers are
mainly composed of type I collagen. Crowded
between the collagen fibers are rows of
fibroblasts, fiber-forming cells, that generate
the fibers.
CLINICAL RELEVANCE
• Marfan Syndrome: defect in elastic fiber
synthesis; reduced elasticity in skin and lungs,
skeletal defects (bones are longer and thinner
than usual), cardiovascular complications
(aneurism, valve prolapse)
 Ehlers-Danlos Syndromes

• 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.

Source Undetermined
ASSIGNMENT/STUDY TASK
• What are the Stains Used in Dense and Loose
Connenctive Tissue Study
• What are the types of Glycosaminoglycans and
their spread in tissues
• What are clinical scenarios associated with
connective tissue disorders.
THANK YOU