Connective tissue,
Bone and Cartilage
CONNECTIVE TISSUE
• Connective tissue consists of cells separated by varying
  amounts of extracellular substance.

• In connective tissues cells typically account for only a
  small fraction of the tissue volume.

• The extracellular substance consists of fibres which are
  embedded in ground substance containing tissue fluid.

• Fibres in connective tissue can be divided into three
  types: collagen fibres, reticular fibres and elastic fibres
Collagen fibres
•   Collagen fibres are the dominant fibre type in most connective tissues. The
    primary function of collagen fibres is to add strength to the connective tissue.

•   The thickness of the fibres varies from ~ 1 to 10 µm. Each of these fibrils is
    composed of microfibrils, which are only visible using electron microscopy.

•   Microfibrils are assemblies of tropocollagen, which, in turn, is an spiral-like
    assembly of three collagen molecules (triple helix).

•   There are many different tropocollagen types around (currently named type I to
    XXI). These types differ in their content of the amino acids hydroxyproline and
    hydroxylysine. They also differ in the amount of carbohydrates attached to the
    collagen molecules.

•   The different types of tropocollagen give the fibres the structural and functional
    features which are appropriate for the organ in which the fibres are found.

•   Types I, II and III are the major fibre-forming tropocollagens. Tropocollagen
    type IV is an important structural component of the basal lamina.
Ct, bone, cartilage
Reticular fibres
• Reticular fibres are very delicate and form fine networks instead
  of thick bundles.
• They are usually not visible in histological sections but can be
  demonstrated by using special stains. For example, in silver
  stained sections reticular fibres look like fine, black threads -
  coarse collagen fibres appear reddish brown in the same type of
  preparation.
• Because of their different staining characteristics, reticular fibres
  were initially thought to be completely different from collagen
  fibres.
• We now know that reticular fibres consist of collagen - although
  the main type of tropocollagen found in reticular fibres, type III,
  is different from that of the coarse collagen fibres.

• Reticular fibres give support to individual cells, for example, in
  Liver, spleen and lymph node.
Ct, bone, cartilage
Elastic fibres
•   Special stains are necessary to show elastic fibres in tissue sections. Resorcin
    fuchsin is one of these stains, which gives the elastic fibres a dark violet
    colour.

•   Light microscopy does not reveal any substructure in the elastic fibres.
    Electron microscopy shows that elastic fibres consist of individual
    microfibrils, which are embedded in an amorphous matrix. The matrix
    accounts for about 90% of the fibre and is composed of the protein elastin.


•   Typically elastic fibres will appear as fine, dark violet and gently undulating
    fibres in the tissue. Elastic fibres can form membranes - not unlike the
    collagen membrane in the basal lamina of epithelia.

•   Elastic fibres can be stretched to about 150% of their original length. They
    resume their original length if the tensile forces applied to the elastic fibres
    are relaxed.
Ct, bone, cartilage
Ct, bone, cartilage
Ground substance
•   Ground substance is found in all cavities and clefts between the fibres and cells
    of connective tissues.

•   Ground substance is soluble in most of the solvents used to prepare histological
    sections and therefore not visible in ordinary sections

•   Water, salts and other low molecular substances are contained within the ground
    substance, but its main structural constituent are proteoglycans.

•   Proteoglycans are responsible for the highly viscous character of the ground
    substance.

•   Proteoglycans consist of proteins (~5%) and polysaccharide chains (~95%),
    which are covalently linked to each other. The polysaccharide chains belong to
    one of the five types of glycosaminoglycans, which form the bulk of the
    polysaccharides in the ground substance.
• Hyaluronan (or hyaluronic acid) is the dominant
  glycosaminoglycan in connective tissues:
   – Hyaluronan serves as a "backbone" for the assembly of other
     glycosaminoglycans in connective and skeletal tissue
   – major component of the synovial fluid, which fills joint cavities,
     and the vitreous body of the eye.


• Other four major glycosaminoglycans are chondroitin
  sulfate, dermatan sulfate, keratan sulfate and heparan
  sulfate

• The components of the ground substance, collagen, elastic
  and reticular fibres are synthesised by cells of the
  connective tissues, the fibrocytes
Connective Tissue Cells
• Connective tissue cells are usually divided
  into two groups based on their ability to
  move within the connective tissue.
• Fibrocytes (or fibroblasts) and fat cells are
  fixed cells.
• Macrophages, monocytes, lymphocytes,
  plasma cells, eosinophils and mast cells are
  wandering cells.
Fibrocytes
•   Fibrocytes are the most common cell type in connective tissues. They are the
    "true" connective tissue cells.

•   Usually only their oval, sometimes flattened nuclei are visible in LM
    sections.

•   The cytoplasm of a resting (i.e. inactive) fibrocyte does not contain many
    organelles.

•   This situation changes if the fibrocytes are stimulated, for example, by
    damage to the surrounding tissue. In this case the fibrocyte is transformed
    into a fibroblast, which contains large amounts of the organelles which are
    necessary for the synthesis and excretion of proteins needed to repair the
    tissue damage

•   Fibrocytes do not usually leave the connective tissue. They are, however,
    able to perform amoeboid movement.
Ct, bone, cartilage
Reticular cells: Reticular cells are usually larger than an average fibrocyte. They
are the "fibrocytes" of reticular connective tissue and form a network of reticular
fibres

Adipocytes: Fat cells or adipocytes are fixed cells in loose connective tissue. Their
main function is the storage of lipids

Macrophages: Macrophages arise from precursor cells called monocytes.

    •Monocytes originate in the bone marrow from where they are released into the
    blood stream.
    •They are actively mobile and leave the blood stream to enter connective
    tissues, where they differentiate into macrophages.
    •Macrophages change their appearance depending on the demand for
    phagocytotic activity

Mast cells: like macrophages, lymphocytes and eosinophils - in demand when
something goes wrong in the connective tissue
Ct, bone, cartilage
Connective Tissue Types
• Loose connective tissue and dense connective tissues

• These two tissues are distinguished according to the relative
  amounts of fibres they contain.

• Dense connective tissues are completely dominated by fibres.
  They are subdivided according to the spatial arrangement of the
  fibres in the tissue.
   – Dense irregular connective tissue the fibres do not show a clear orientation
     within the tissue but instead form a densely woven three-dimensional
     network, example is the dermis of the skin.
   – Regular dense connective tissue if the fibres run parallel to each other,
     examples are tendons, ligaments and the fasciae and aponeuroses of
     muscles

• Loose connective tissue : It is best understood as a kind of
  generalised connective tissue in which all connective tissue cell
  types may occur
Ct, bone, cartilage
Bone
Gross
• 'Compact or cortical bone’ is usually thick dense
  bone that forms the outer shell covering almost all
  bones in the body.
• ‘Cancellous bone’- the inner aspect consists of
  bone that is not as dense as cortical bone and has a
  honeycomb appearance
BONE
Ct, bone, cartilage
OSSEOUS TISSUE
•   CELLS:
    – osteoblasts - take origin from poorly differentiated mesenchymal cells; reside in the internal
      layer of periosteum, during bone development osteoblasts are on the periosteal surface and
      around interosseous blood vessels; these cells are cuboidal, columnar and polygonal in shape,
      have a well-developed rough endoplasmic reticulum
        function - production of bone intercellular matter

    – osteocytes - a mature form of osteoblasts, they lie in lacunae within a bone and extend
      protoplasmic processes into small canaliculi in the intercellular matrix
      function - weak secretion of bone intercellular matter components

    – osteoclasts - macrophages of bone tissue, blood monocytes being their precursors; large
      multinucleated cells; a zone of cytoplasm adjacent to osseous surface is referred to as ruffled
      border, multiple cytoplasmic processes and lysosomes are found here
      function - destruction and resorption of bone fibers and ground substance

•   INTERCELLULAR MATTER
    – FIBERS: collagen fibers
    – GROUND (AMORPHOUS) SUBSTANCE: the predominant component is calcium
      phosphate mostly in the form of hydroxyapatite crystals and some in amorphous state; small
      amounts of magnesium phosphate and very scarce glycosaminoglycans and proteoglycans are
      also found
Ct, bone, cartilage
Types of the bone
• Fibrous (immature) bone
   – ossein fiber orientation is random
   – cells are embedded into the intercellular matrix, reside on bone surfaces and
     around vessels penetrating the bone

• Lamellar (mature) bone
   – ossein fibers are regularly arranged, forming bone lamellae
   – fibers in a lamella have identical orientation
   – cells reside between lamellae in specialized lacunae as well as around
     penetrating vessels
   – cells have processes, which they can connect to each other by
   – apart from osseous lamellae in lamellar bone there are special structures -
     osteons
   – osteon forms around a vessel, therefore there is always a vessel in the
     center of osteon, and circular lamellae with some cells scattered in-between
     are present around a vessel
   – an osseous channel through which a blood vessel traverses is referred to as
     Haversian canal
Ct, bone, cartilage
• PERIOSTEUM: has 2 layers:
     external - fibrous; made of dense irregular connective tissue
     internal - cellular (osteogenic); contains many osteoblasts
                and blood vessels, some osteocytes as well
   functions: bone nutrition, longitudinal and transverse growth of bone, its
             regeneration

• ENDOSTEUM
  a lining covering a bone from the marrow side, made of loose
  irregular connective tissue with osteoblasts and osteoclasts in
  addition to more common cell types of this tissue
Ct, bone, cartilage
CARTILAGE
CARTILAGINOUS TISSUE
•   CELLS
    – Chondroblasts - less differentiated cartilage cells, originate from non-differentiated
      mesenchyme; have a flattened shape; a basophilic cytoplasm;

    – function - elaboration of cartilage intercellular matter; under certain circumstances
      chondroblasts are capable of producing matrix-degrading enzymes - collagenase,
      elastase, hyaluronidase

    – reside in the internal layer of periosteum and in the depth of matrix - within lacunes
      chondroblasts mature into chondrocytes


    – Chondrocytes - differentiated cartilage cells; of round or angular shapes, with
      advancing cellular age chondrocytes progressively lose their rough endoplasmic
      reticulum;
      function - elaboration of cartilage intercellular matter; under certain circumstances
      chondroblasts are capable of producing matrix-degrading enzymes - collagenase,
      elastase, hyaluronidase

    – reside in the depth of matrix - within minute special cavities lacunes sometimes the
      number of cartilage cells in one lacune is more than one, it is the consequence of cell
      division;
Hyaline cartilage

• Localization: trachea
  and bronchi, articular
  surfaces, larynx,
  costosternal junctions

• Intercellular matter
   – Fibers: collagen fibers
   – Ground substance:
     glycosaminoglycans
     and proteoglycans
Ct, bone, cartilage
Elastic cartilage
•Localization: pinna of the ear, corniculate and cuneiform
cartilages of the larynx, nasal cartilages
•Intercellular matter
    •Fibers: elastic and collagen fibers
    •Ground substance: glycosaminoglycans and proteoglycans

Fibrous cartilage

•Localization: attachments of tendons and ligaments to hyaline
cartilage, intervertebral disks, semimovable joints, and symphysis
•Intercellular matter
    •Fibers: parallel fascicles of collagen fibers; more fibers than in
    other types of cartilage
    •Ground (amorphous) substance: glycosaminoglycans and
    proteoglycans
Ct, bone, cartilage
Ct, bone, cartilage

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Ct, bone, cartilage

  • 2. CONNECTIVE TISSUE • Connective tissue consists of cells separated by varying amounts of extracellular substance. • In connective tissues cells typically account for only a small fraction of the tissue volume. • The extracellular substance consists of fibres which are embedded in ground substance containing tissue fluid. • Fibres in connective tissue can be divided into three types: collagen fibres, reticular fibres and elastic fibres
  • 3. Collagen fibres • Collagen fibres are the dominant fibre type in most connective tissues. The primary function of collagen fibres is to add strength to the connective tissue. • The thickness of the fibres varies from ~ 1 to 10 µm. Each of these fibrils is composed of microfibrils, which are only visible using electron microscopy. • Microfibrils are assemblies of tropocollagen, which, in turn, is an spiral-like assembly of three collagen molecules (triple helix). • There are many different tropocollagen types around (currently named type I to XXI). These types differ in their content of the amino acids hydroxyproline and hydroxylysine. They also differ in the amount of carbohydrates attached to the collagen molecules. • The different types of tropocollagen give the fibres the structural and functional features which are appropriate for the organ in which the fibres are found. • Types I, II and III are the major fibre-forming tropocollagens. Tropocollagen type IV is an important structural component of the basal lamina.
  • 5. Reticular fibres • Reticular fibres are very delicate and form fine networks instead of thick bundles. • They are usually not visible in histological sections but can be demonstrated by using special stains. For example, in silver stained sections reticular fibres look like fine, black threads - coarse collagen fibres appear reddish brown in the same type of preparation. • Because of their different staining characteristics, reticular fibres were initially thought to be completely different from collagen fibres. • We now know that reticular fibres consist of collagen - although the main type of tropocollagen found in reticular fibres, type III, is different from that of the coarse collagen fibres. • Reticular fibres give support to individual cells, for example, in Liver, spleen and lymph node.
  • 7. Elastic fibres • Special stains are necessary to show elastic fibres in tissue sections. Resorcin fuchsin is one of these stains, which gives the elastic fibres a dark violet colour. • Light microscopy does not reveal any substructure in the elastic fibres. Electron microscopy shows that elastic fibres consist of individual microfibrils, which are embedded in an amorphous matrix. The matrix accounts for about 90% of the fibre and is composed of the protein elastin. • Typically elastic fibres will appear as fine, dark violet and gently undulating fibres in the tissue. Elastic fibres can form membranes - not unlike the collagen membrane in the basal lamina of epithelia. • Elastic fibres can be stretched to about 150% of their original length. They resume their original length if the tensile forces applied to the elastic fibres are relaxed.
  • 10. Ground substance • Ground substance is found in all cavities and clefts between the fibres and cells of connective tissues. • Ground substance is soluble in most of the solvents used to prepare histological sections and therefore not visible in ordinary sections • Water, salts and other low molecular substances are contained within the ground substance, but its main structural constituent are proteoglycans. • Proteoglycans are responsible for the highly viscous character of the ground substance. • Proteoglycans consist of proteins (~5%) and polysaccharide chains (~95%), which are covalently linked to each other. The polysaccharide chains belong to one of the five types of glycosaminoglycans, which form the bulk of the polysaccharides in the ground substance.
  • 11. • Hyaluronan (or hyaluronic acid) is the dominant glycosaminoglycan in connective tissues: – Hyaluronan serves as a "backbone" for the assembly of other glycosaminoglycans in connective and skeletal tissue – major component of the synovial fluid, which fills joint cavities, and the vitreous body of the eye. • Other four major glycosaminoglycans are chondroitin sulfate, dermatan sulfate, keratan sulfate and heparan sulfate • The components of the ground substance, collagen, elastic and reticular fibres are synthesised by cells of the connective tissues, the fibrocytes
  • 12. Connective Tissue Cells • Connective tissue cells are usually divided into two groups based on their ability to move within the connective tissue. • Fibrocytes (or fibroblasts) and fat cells are fixed cells. • Macrophages, monocytes, lymphocytes, plasma cells, eosinophils and mast cells are wandering cells.
  • 13. Fibrocytes • Fibrocytes are the most common cell type in connective tissues. They are the "true" connective tissue cells. • Usually only their oval, sometimes flattened nuclei are visible in LM sections. • The cytoplasm of a resting (i.e. inactive) fibrocyte does not contain many organelles. • This situation changes if the fibrocytes are stimulated, for example, by damage to the surrounding tissue. In this case the fibrocyte is transformed into a fibroblast, which contains large amounts of the organelles which are necessary for the synthesis and excretion of proteins needed to repair the tissue damage • Fibrocytes do not usually leave the connective tissue. They are, however, able to perform amoeboid movement.
  • 15. Reticular cells: Reticular cells are usually larger than an average fibrocyte. They are the "fibrocytes" of reticular connective tissue and form a network of reticular fibres Adipocytes: Fat cells or adipocytes are fixed cells in loose connective tissue. Their main function is the storage of lipids Macrophages: Macrophages arise from precursor cells called monocytes. •Monocytes originate in the bone marrow from where they are released into the blood stream. •They are actively mobile and leave the blood stream to enter connective tissues, where they differentiate into macrophages. •Macrophages change their appearance depending on the demand for phagocytotic activity Mast cells: like macrophages, lymphocytes and eosinophils - in demand when something goes wrong in the connective tissue
  • 17. Connective Tissue Types • Loose connective tissue and dense connective tissues • These two tissues are distinguished according to the relative amounts of fibres they contain. • Dense connective tissues are completely dominated by fibres. They are subdivided according to the spatial arrangement of the fibres in the tissue. – Dense irregular connective tissue the fibres do not show a clear orientation within the tissue but instead form a densely woven three-dimensional network, example is the dermis of the skin. – Regular dense connective tissue if the fibres run parallel to each other, examples are tendons, ligaments and the fasciae and aponeuroses of muscles • Loose connective tissue : It is best understood as a kind of generalised connective tissue in which all connective tissue cell types may occur
  • 19. Bone
  • 20. Gross • 'Compact or cortical bone’ is usually thick dense bone that forms the outer shell covering almost all bones in the body. • ‘Cancellous bone’- the inner aspect consists of bone that is not as dense as cortical bone and has a honeycomb appearance
  • 21. BONE
  • 23. OSSEOUS TISSUE • CELLS: – osteoblasts - take origin from poorly differentiated mesenchymal cells; reside in the internal layer of periosteum, during bone development osteoblasts are on the periosteal surface and around interosseous blood vessels; these cells are cuboidal, columnar and polygonal in shape, have a well-developed rough endoplasmic reticulum function - production of bone intercellular matter – osteocytes - a mature form of osteoblasts, they lie in lacunae within a bone and extend protoplasmic processes into small canaliculi in the intercellular matrix function - weak secretion of bone intercellular matter components – osteoclasts - macrophages of bone tissue, blood monocytes being their precursors; large multinucleated cells; a zone of cytoplasm adjacent to osseous surface is referred to as ruffled border, multiple cytoplasmic processes and lysosomes are found here function - destruction and resorption of bone fibers and ground substance • INTERCELLULAR MATTER – FIBERS: collagen fibers – GROUND (AMORPHOUS) SUBSTANCE: the predominant component is calcium phosphate mostly in the form of hydroxyapatite crystals and some in amorphous state; small amounts of magnesium phosphate and very scarce glycosaminoglycans and proteoglycans are also found
  • 25. Types of the bone • Fibrous (immature) bone – ossein fiber orientation is random – cells are embedded into the intercellular matrix, reside on bone surfaces and around vessels penetrating the bone • Lamellar (mature) bone – ossein fibers are regularly arranged, forming bone lamellae – fibers in a lamella have identical orientation – cells reside between lamellae in specialized lacunae as well as around penetrating vessels – cells have processes, which they can connect to each other by – apart from osseous lamellae in lamellar bone there are special structures - osteons – osteon forms around a vessel, therefore there is always a vessel in the center of osteon, and circular lamellae with some cells scattered in-between are present around a vessel – an osseous channel through which a blood vessel traverses is referred to as Haversian canal
  • 27. • PERIOSTEUM: has 2 layers: external - fibrous; made of dense irregular connective tissue internal - cellular (osteogenic); contains many osteoblasts and blood vessels, some osteocytes as well functions: bone nutrition, longitudinal and transverse growth of bone, its regeneration • ENDOSTEUM a lining covering a bone from the marrow side, made of loose irregular connective tissue with osteoblasts and osteoclasts in addition to more common cell types of this tissue
  • 30. CARTILAGINOUS TISSUE • CELLS – Chondroblasts - less differentiated cartilage cells, originate from non-differentiated mesenchyme; have a flattened shape; a basophilic cytoplasm; – function - elaboration of cartilage intercellular matter; under certain circumstances chondroblasts are capable of producing matrix-degrading enzymes - collagenase, elastase, hyaluronidase – reside in the internal layer of periosteum and in the depth of matrix - within lacunes chondroblasts mature into chondrocytes – Chondrocytes - differentiated cartilage cells; of round or angular shapes, with advancing cellular age chondrocytes progressively lose their rough endoplasmic reticulum; function - elaboration of cartilage intercellular matter; under certain circumstances chondroblasts are capable of producing matrix-degrading enzymes - collagenase, elastase, hyaluronidase – reside in the depth of matrix - within minute special cavities lacunes sometimes the number of cartilage cells in one lacune is more than one, it is the consequence of cell division;
  • 31. Hyaline cartilage • Localization: trachea and bronchi, articular surfaces, larynx, costosternal junctions • Intercellular matter – Fibers: collagen fibers – Ground substance: glycosaminoglycans and proteoglycans
  • 33. Elastic cartilage •Localization: pinna of the ear, corniculate and cuneiform cartilages of the larynx, nasal cartilages •Intercellular matter •Fibers: elastic and collagen fibers •Ground substance: glycosaminoglycans and proteoglycans Fibrous cartilage •Localization: attachments of tendons and ligaments to hyaline cartilage, intervertebral disks, semimovable joints, and symphysis •Intercellular matter •Fibers: parallel fascicles of collagen fibers; more fibers than in other types of cartilage •Ground (amorphous) substance: glycosaminoglycans and proteoglycans