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GENERAL CHARACTERISTICS
Like other CT, it consists of cells, fibers and ground substance or matrix. Cavities called lacunae containing the bone cells or osteocytes are uniformly spaced in the matrix. Radiating canaliculi found in the bony lacunae lodge in the cytoplasmic processes of the cells. Bone is vascular tissue; nutrition of bone is carried through the continuous communication of bone canaliculi with the blood vessels located in and outside the bone. The dense bone matrix is devoid of blood vessels.
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 Bones are invested on the external surface by the periosteum, a layer of specialized CT endowed with osteogenic or bone-forming potentialities. The internal layer termed the endosteum, which have both osteogenic and hemopoietic (blood-forming) abilities, is a layer of flattened osteoprogenitor cells and small amount of CT. All the cavities of bone including the Haversian canals and the marrow spaces within spongy bone are lined by endosteum. There are certain areas where a periosteal covering is lacking such as the ends of long bones that are covered by articular cartilage, where tendons and ligaments insert into the bone or on the surface of the patella, and in sesamoid bones formed within tendons.
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FUNCTIONS:
Its extracellular components are calcified making it rigid, hard and brittle, thus making it suited for its mechanical function of skeletal support. Provides for the internal support of the body and for attachment of muscles and tendons essential for locomotion. Protects vital organs of the cranial and thoracic cavities. Encloses the bloodforming elements of the bone marrow. Serves as site for the storage of calcium, phosphates and ions.
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GROUND SUBSTANCE
The bony matrix is composed of calcified substance made up of organic and inorganic elements deposited in lamellae. The ORGANIC elements are composed of proteoglycan aggregates and structural glycoproteins responsible for promoting calcification of bone matrix. There are proteins found only in bone, the synthesis of which are stimulated by Vit. D: osteocalcin, osteopontin, and bone sialoprotein. Type I collagen fibers with lower sulfate content than that of the hyaline matrix render the bone matrix acidophilic in staining.
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 The INORGANIC matter of bone consists of calcium, magnesium and sodium phosphates, very similar but not identical to the mineral hydroxyapatite Ca10(PO4)6(OH)2. The hydration shell of hydroxyapatite facilitates ion exchange between the crystal and body fluids.
Demineralized bone retains its gross form but loses its hardness, yet remaining tough and flexible. Removal of the matrix organic content leaves bone with its original shape, but extremely brittle when handled.
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1.Osteoprogenitor cells- are relatively undifferentiated cells having the capacity for further structural and functional specialization. They have pale-staining oval or elongated nuclei and an inconspicuous acidophilic or faintly basophilic cytoplasm. They are found on the free bony surfaces, endosteum, periosteum, lining the Haversian canals and at the epiphyseal plate of growing bones. They are active during the normal growth of bones. In adult life, they maybe activated during healing of fractures or repair of other forms of injury. They undergo division, transforming into osteoblasts (bone-forming cells), or unite giving rise to osteoclasts (bone-destroying cells).
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2.Osteoblasts- responsible for formation of bone matrix and found on the surfaces of developing bones. They are arranged during the active stage in an epithelioid layer of cuboidal or low columnar cells connected to each other by slender, short processes. The nucleus is often located at the end of the cell farthest from the bony surface. The elongated mitochondria are fairly numerous; the Golgi apparatus is well developed. Owing to its large content of RNPs, the cytoplasm appears intensely basophilic. The cells give a strong histochemical reaction for alkaline phosphatase. The PAS reaction shows small pink-staining granules in the cytoplasm believed to be precursors of the bone matrix.
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 EM reveal structures of cell actively engaged in protein synthesis like an extensive ER, ribosomes, well developed Golgi complex, small lipid droplets, and membrane-limited dense bodies. During matrix synthesis, a layer of new uncalcified matrix (osteoid) forms between the osteoblast layer and old matrix, which later becomes calcified. The rate at which this process (bone apposition) takes place can be measured by the fluorescent antibiotic tetracycline and is of diagnostic importance in diseases such as osteomalacia (impaired mineralization) and osteitis fibrosa cystica (increased osteoclast acitvity). Osteoblasts also participate in bone resorption by secreting enzymes that remove the superficial layer of unmineralized matrix (osteoid), exposing the mineralized matrix to activated osteoclasts. 9

3.Osteocytes- the principal cells of fully formed bone residing in lacunae within the calcified interstitial substance. An osteocyte is essentially an osteoblast that has become surrounded by bony matrix in its development. Under the LM, the nuclear and cytoplasmic characteristics resemble those of the osteoblasts except that the cytoplasm has less affinity to basic dyes and the Golgi region is less conspicuous. The cell body is flattened according to the lacuna that it occupies Slender, numerous processes extend into canaliculi in the surrounding matrix which form gap junctions or nexuses at their sites of contact.
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 Some molecular exchange that takes place between osteocytes and blood vessels can nourish a chain of about 15 cells. Osteocytes play an active role in the release of calcium from bone to blood. Thus they participate in the homeostatic regulation of its concentration in the fluids of the body. Osteoblasts and osteocytes can revert to osteoprogenitor cells. This ability confers them not only a high plasticity to adapt quickly to changing conditions, but also a tendency to escape normal controls of proliferation, causing in adolescents or young adults benign (osteoblastoma, osteoclastoma), or malignant (osteosarcoma) tumors. Bone metastases also occur from breast, lung, kidney and thyroid tumors. 11

4.Osteoclasts- giant, polar, multinucleated cells found in Howships lacunae, associated with areas of bone resorption. They are derived from the granulocyte-macrophage progenitor cells (GM-CSF) of the bone marrow. Osteoclasts are regarded as secretory cells releasing acid hydrolases and pumping H+ ions into the subosteoclastic compartment to digest bone matrix. The nuclei of osteoclasts resemble those of the osteoblasts; the cytoplasm is slightly basophilic when stained with basic dyes. In routine histological sections, it is usually eosinophilic and vacuolated Osteoclast activity is controlled by cytokines (osteoclast stimulating factor) secreted by osteoblast upon stimulation of PTH & calcitonin.
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 In active osteoclasts, the surface facing the bone matrix is folded into irregular, subdivided projections termed ruffled border. Surrounding this is an actinrich clear zone, which is the microenvironment in which bone resorption occurs. In the genetic disease osteopetrosis (dense, heavy marble bones), osteoclasts lack ruffled borders, hence bone resorption is defective.
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2 forms of bones distinguishable with the naked eye: 1.Compact (substantia compacta)- ground and/or decalsified bone Appears as a solid continuous mass containing spaces seen only with the aid of a microscope. Found only in the outer surface of all bones. The shaft (diaphysis) consists of a thick-walled hollow cylinder of compact bone with a central medullary cavity containing bone marrow. The ends of long bones consist mainly of spongy bone covered by a thin, peripheral cortex of compact bone.
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 The epiphysis end of long bones are separated from the diaphysis by a cartilaginous epiphyseal plate. Columns of spongy bone unite this plate to the diaphysis in a region called the metaphysis. Immediately beneath the periosteum at the external surface of bone are several lamellae that extend uninterruptedly around the circumference of the shaft. These constitute the periosteal or external circumferential lamellae. Immediately surrounding the central medullary cavity are several lamellae constituting the endosteal or inner circumferential lamellae.
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 Between these external and internal circumferential lamellae are located the Haversian systems or osteons, composed of concentric lamellae where lacunae are located, and a central Haversian canal. Between the Haversian systems are the interstitial systems, which are angular fragments of lamellar bone of varying size and irregular shape.
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Two categories of vascular channels are distinguished in compact bone on the basis of their orientation and relation to the lamellar structure of the surrounding bone: 1.Haversian canals: longitudinal channels in the centers of the Haversian systems containing 1 or 2 blood vessels. They are connected with one another and communicate with the free surface and marrow cavity through transverse and oblique channels called Volkmanns canals 2.Volkmanns canals: distinguished from the Haversian canals since they are not surrounded by concentrically arranged lamellae but traverse the bone in a direction perpendicular or oblique to the lamellae. They are often larger than Haversian canals. Blood vessels from the periosteum traverse the Volkmanns canals and finally the medullary cavities.
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 The Haversian canals communicate with one another thru the canaliculi. They also communicate with the blood vessels of the periosteum and bone marrow thru the Volkmanns canals. Haversian lamellae contain alternating bright and dark concentric layers that result in the differing orientation of collagen fibers in the successive lamellae. In the flat bones of the skull, the substantia compacta forms on both surfaces thick layers referred to as the inner and outer tables. The layer of spongy bone of varying thickness between these tables constitute the diploe. The periosteum on the outer surface is known as the pericranium, while that on the inner surface is the dura mater.
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In cross-section, compact bone appears as a mosaic of round and angular pieces cemented together.

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2.Cancellous/ spongy bone (substantia spongiosa)cartilage bone developing Found in the diploe of flat bones of the skull and face, in the middle or inner portion of all other bones. It is found only as a thin portion inside the diaphysis of long bones but constitutes a greater part of the epiphysis. Consists of a 3-D lattice of branching bony spicules or trabeculae enclosing a system of intercommunicating spaces that are occupied by bone marrow. These bony spicules are made up of layers of homogenous bone matrix consisting of organic and inorganic elements.
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 The trabeculae are relatively thin, and are not usually permeated by blood vessels. The numerous small, communicating marrow cavities or intrabecular spaces vary greatly in size and shape.

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 They are lined by a squamous cellular membrane called endosteum. Hemopoietic, myeloid tissue and some fat cells occupy these cavities. There are usually no Haversian systems. The osteocytes are embedded in the trabeculae The cells are ovoid in shape with fine cytoplasmic projections.

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 Each osteocyte is lodged in a lacuna which possess minute radiating canaliculi. The cytoplasmic projections of the osteocytes extend into these small canaliculi. The bone cells are nourished by diffusion from the surface thru the minute bony canaliculi that interconnect lacunae and extend to the surface. In prenatal spongy bone, the lamellae are indistinct since the osteo-collagenous fibers form an irregular network. This is characteristic of rapid bone development and is referred to as woven bone. Isolated patches of such bone occur in the adult and during repair of fractures.
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 Bone can be formed by: a.Intramembranous ossification- direct mineralization of matrix secreted by osteoblasts b.Endochondral ossification- deposition of bone matrix on a pre-existing cartilage. In both processes, the bone tissue that first appears is primary or woven bone, soon replaced by definitive lamellar or secondary bone. Bone synthesis and remodeling occurs throughout life, although rates of change maybe considerably slower in adults.

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1.Intramembranous Ossification- membrane bone Bone formation takes place directly in primitive CT mesenchyme. This occurs in the development of the flat bones of the skull and face, referred to as membrane or dermal bones. In the mesenchymal condensation layer, a primary ossification center appears. Groups of cells differentiate into osteoblasts which secrete fine collagenous fibers (osteogenic fibers of Sharpey) that form bundles which anastomose into a network and make up primary or woven bone. 26

 The trabeculae continue to thicken at the expense of the intervening CT until the spaces around the blood vessels are obliterated in areas of the primary spongiosa destined to become compact bone. The collagen fibers & the osteocytes in lacunae become arranged in regularly concentric cylinders resembling lamellar bone. The osssification centers grow radially and fuse together, replacing the original CT.
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 Osteoblasts form a soft bone precursor material termed osteoid bone (red-stained) grown by apposition. The cells inside the osteoid lamellae have entrapped themselves during osteogenesis and have differentiated into osteocytes. The bone matrix becomes a site of deposition of calcium phosphate, trapping the osteocytes in lacunae which are uniformly distributed but oriented at random. The canaliculi are formed by deposition of matrix around the cell processes of the osteoblasts. 28

 The richly vascularized mesenchymal tissue between the primary bony lamellae (spongy bone trabeculae) is called primary bone marrow. Later, secondary bone marrow, which forms the blood cells, will replace it. Multinucleated giant cells or osteoclasts degrade bone tissue (bone resorption) and phagocytose the products of this degradation. When they degrade osteoid material, they create small resorption bays, termed Howship lacunae
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 In areas where spongy bone will persist, the intervening vascular CT is gradually transformed into hemopoietic tissue. The remaining unossified portion gives rise to the periosteum and endosteum. The fontanelles of newborn infants also correspond to CT that are not yet ossified. Bone destruction by osteoclasts is slower at the periphery of the bone, and results in the formation of a plate of compact bone enclosing the spongy bone (diploe). Being thin, Haversian systems are rare.
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2.Endochondral or Intracartilaginous Ossificationcartilage bone developing Replacement of a cartilage model by bone. These cartilage, substitute, or replacement bones constitute the bones of the entire skeletal system except the flat bones of the skull and face that develop through intramembranous ossification.

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a.Fetal hyaline cartilage formation- arising from a mesenchymal blastema, it assumes the shape of the future bone. b.Appearance of the ossification centers- in long bones, 3 centers appear: in the middle of the shaft (diaphyseal or 10 center of ossification); and at the center of each epiphyses or 2 ends of the long bone (20 centers of ossification).
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c.Formation of the periosteal band or bone collarosteogenic properties of the cells in the perichondrium are activated. A thin layer of bone is deposited around the middle of the shaft d.Absorption of cartilage matrix- occurs faster among cells of the same row or column, and accomplished through the action of alkaline phosphatase secreted by enlarging cartilage cells. Lacunae enlarge, while the intervening cartilage matrix is gradually reduced to thin septa and irregularly shaped spicules. Due to absorption of cartilage matrix, parallel cavities (primitive or 10 marrow cavities) form.

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1) Zone of proliferation- frequent mitotic division provides for continuous elongation of the columns. 2) Zone of maturation- cells are no longer dividing but gradually enlarge. 3) Zone of hypertrophy- appears bluish gray; cells appear big and vacuolated; because this is also a zone of calcification of remaining strips of cartilage matrix, the area is called the zone of provisional calcification. A lag in calcification occurs under conditions of local failure in calcium or phosphate supply. 4. Zone of cell degeneration- cells are degenerating and their enlarged lacunae are invaded by capillary loops and primitive osteogenic cells.
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e.Appearance of periosteal buds- from the inner layer of fetal periosteum, buds of fibrous tissue containing blood vessels, osteogenic and hemopoietic elements appear towards the 10 marrow cavities, and invade the cartilage matrix. Some osteogenic cells form the endosteum, while the rest of the tissue constitute the fetal bone marrow or myeloid tissue.

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f. Primary bone depositionbony or osseous tissue is deposited upon the strips of remaining cartilage matrix by the osteoblasts brought in by the buds, resulting in 10 bone that appear in the form of irregular trabeculae between 10 marrow cavities. Some cells become osteocytes, while the rest continue laying down bony tissue resulting in the formation of wider strips of 10 bone.
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f. Primary bone destructionosteoclasts derived from buds start to function through their proteolytic enyme activity. Since bone destruction is faster than bone formation in shafts of long bones, one large medullary cavity (20 marrow cavity) lined by the endosteum is formed. It contains the primitive bone marrow. g.Periosteal Ossification- deposits of parallel & concentric bony tissue by the osteoblasts appear beneath the periosteum, resulting in widening of the original periosteal band. Within the compact bone matrix, there are formed longitudinal cavities that are parallel to the central medullary cavity.

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 These cavities become Haversian canals lined by endosteum; the concentric bony lamellae complete the Haversian systems. They are not permanent since they are continually destroyed by osteoclasts. The areas of bony matrix between these Haversian systems constitute the interstitial or ground lamellae. The bigger canals having a horizontal direction are formed by the passage inwards of the periosteal buds. They are not surrounded by concentric lamellae and become Volkmanns canals. These cavities are invaded and lined by osteoblasts which start laying down concentric bony lamellae until the lumen is reduced. Through this process, bones acquire growth in width. 41

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h.Epiphyseal Ossification- at about the time of birth, 20 centers of ossification appear in both ends of most long bones. The sequence of changes in the cartilage of these centers is identical with that observed in the diaphysis. The epihyseal plate or disc is cartilage that remains over the fine end as articular cartilage and as a plate between the diaphysis and epiphyses. Bone destruction is not marked, hence no single marrow cavity forms but rather thin and anastomosing bone trabeculae with small marrow cavities of the spongy or cancellous bone. Around this spongy bone, a thin plate of compact bone is laid down by the endosteum. Growth in length of the femur takes place mainly at the distal epiphyses; growth of the tibia takes place mainly at the proximal end (important in orthopedic surgery and radiology). 43

i. Periosteal and endosteal lamellae formation- the inner layer of the periosteum begin to deposit successive uninterrupted bony lamellae constituting the periosteal or external circumferential lamellae. The endosteum also lay down upon the wall of the central medullary cavity the endosteal or internal circumferential lamellae. Since the osteoclasts are no longer active, the lamellae appear continuous. Through the activity of the periosteum, increase in width of the bone is achieved. Through the enlargement of the chondrocytes towards the epiphyses, the epiphyseal line is pushed farther resulting in the lengthening of the bone. 44

 Since length of bone is limited to its epiphyseal plates, injury to this region may result in serious growth impairment. In cases of the retarded growth of one leg due to neurovascular disturbances as in poliomyelitis, the orthopedic surgeon can select the appropriate time and site for a surgical obliteration of the epiphyses in the normal leg. There are existing knowledge of the normal rates of growth at the epiphyseal plates and the times of their normal closure. By such procedure, growth of the normal leg maybe retarded just enough for the slow growing leg to catch up, thus achieving an equalization of leg length. The shape of bone is maintained by a continual remodeling of its surface, involving periosteal deposition in some areas and bone absorption in others. 45

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sites where 2 or more components of the skeleton meet. Types of joints: 1. Synostosis- bones are united by bone tissue and no movement takes place (skull bones) 2. Synchondrosis- bones are joined by hyaline cartilage (epiphyseal plates, 1st rib to sternum) 3. Syndesmosis- bones joined by interosseous ligament of dense CT (pubic symphysis) These CT structures permit varying degrees of movement between adjoining bones. Joint capsules consist of 2 distinct layers: a. Inner synovial layer- cellular and secretes a colorless, viscid fluid of the joint cavity. It is thrown into marked folds, containing blood vessels and which project into the joint cavity. With age, these folds increase in size and number. b. An external layer of dense CT consisting of collagenous fibers and fibroblasts, macrophages, blood vessels, and many lymphatics. c. Diarthrosis- ligaments and a capsule of CT unite long bones at their points of contact.
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Histologic structure of the synovial membrane, with its lining cells in epithelioid arrangement. There is no basal lamina between the lining cells and the underlying connective tissue. This tissue is rich in blood capillaries and contains a variable number of adipose cells (AD).

1.Primary Osteoporosis- critical loss of bone mass associated with growth hormone deficiency (senile) or estrogen (postmenopausal). Decreased estrogen levels result in increased secretion of IL-1 (a potent stimulator of osteoclasts) from monocytes. 2.Pagets disease- characterized by uncontrolled osteoclast activity, causing widespread bone resorption followed by intense osteoblast activity, producing woven bone that fills in the erosion. The net effect is an increase in bone mass that is unsound because the woven bone persists.

3.Osteomalacia and rickets are a result of Vit. D deficiency, affecting the phosphate concentration in the blood plasma, and results to deficient calcification or failure of osteoid tissue to calcify.  Sources include dietary intake and production by skin keratinocytes stimulated by UV light. The vitamin is hydroxylated in the liver and kidney, required for the absorption of calcium and phosphates during osteoid mineralization.  Physical signs of osteomalacia in adults include bowed legs, increased tendency to fracture, and scoliotic deformity of the vertebral column.  Physical signs of rickets include craniotabes (elastic recoil of the skull upon compression), rachitic rosary (excess osteoid at the costochondral junction), and pigeon-breast deformity ( anterior protrusion of sternum)
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4.Scurvy- results from Vit C deficiency, necessary for the hydroxylation of proline and lysine amino acids during collagen synthesis, causing inability of tissues that differentiate from mesenchyme to produce fibers and ground substance. This causes a destruction of osteocollagenous fibers and a diminished production of organic matrix in bones.  Physical signs include: poor bone growth and poor fracture repair due to lack of collagen within osteoid; hemorrhages in the skin (purpura), gingival mucosa, and joints due to weakened tunica media of blood vessels. 5.Vitamin A- an excess causes premature closure of the epiphyseal plate, resulting in a person of small stature. In Vit. A deficiency, there is a diminution in the rate of skeletal growth and interference with the process of remodeling together with the balance between bone deposition and erosion.
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 The growth and maintenance of bones is greatly influenced by hormones: the growth hormone of the anterior pituitary is essential for normal bone growth: its deficiency leads to dwarfism; excessive production leads to gigantism. In adults, increase in growth hormone causes long bones to become very thick, resulting to acromegaly. Parathyroid hormone regulates bone resorption controlling the release of calcium in the blood; thyrocalcitonin inhibits resorption The appearance and closure of epiphyseal ossification centers are related to the production of sex hormones by the gonads. Sex hormone differences cause taller stature in males due to longer existence of functional epiphyseal cartilages.

 Bone formation involves the interaction of multiple regulatory substances: a.Bone morphogenetic proteins (BMP) that induce endochondral bone formation, the glycoprotein b.Osteogenein that initiates ectopic bone formation c.Transforming growth factor-F (TGF-F), the peptide autocrine factor that stimulate synthesis of matrix components by chondrocytes and osteoblasts. Despite its hardness, the internal structure of bone can be remodeled according to the various stresses to which it is subjected, e.g., orthodontic appliances produce lateral pressures that can modify teeth positions in the jawbone.
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 After a fracture, there is a resulting hemorrhage from torn vessels and clotting. Proliferating fibroblasts and capillaries invade the clot, forming granulation tissue or the procallus. This becomes dense fibrous tissue, transforming later into cartilage. This temporary callus unites the fractured bones. Osteoblasts develop from the periosteum and endosteum, replacing progressively the cartilage of the temporary callus similar to endochondral ossification. The initially spongy bony callus undergoes reorganization into compact bone & excess bone is resorbed.
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