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Connective Tissue Part II

Developing Bone

1. Examine the slide, Developing Intramembranous Bone (page 81). There are essentially 3 divisions in this slide that resemble strips of pink ribbon. The dark peripheral ribbon at the superior surface is periosteum, an irregular connetive tissue. In this tissue osteoprogenitor cells may arise from deep fibroblasts. The fibroblasts are distinguished from the collagen matrix by their purple nuclei. You might see one or two strips of fibroblasts. As you travel to the edge of the bright pink layer that resembles Swiss cheese, you will see numerous purple cells at the periphery. These cells are osteoblasts, the progenitors of osteocytes (true bone cells). The pink matrix itself is osteoid, a pre-bone matrix rich in collagen and in the process of obtaining calcium phosphate crystals (hydroxyapatite) that will ultimately give rigidity to bone. Note that the bone is forming columns called trabeculae that will make the framework for spongy bone. Within the trabeculae are osteocytes contained within their lacunae. For all purposes, they will not become mature osteocytes until the bone calcifies. The inferior pink strip that resembles loose connective tissue is embryonic connective tissue rich in blood vessels. Again, fibroblasts may be seen here, along with red blood cells. Draw what you see in intramembranous bone formation.

2. Examine the slide Bone, Developing Cartilage Type. This is a small developing bone. The tip of the bone is where the joint will form. Turn the slide so that the print on its identification is upside down and hold the slide up to the light before placing it on your microscope. Note the faint pink arch near the superior region of the tissue. We will examine this arch in more detail. Now turn the slide right side up, so that the print may be read correctly from the microscope stage and place your slide on the microscope. Examine the slide under low power and try to find the arch. Note that the tissue above the arch is cartilage; below the arch the tissue is osteoid. Lets look at the cartilage tissue above the arch first. You may recognize this tissue as hyaline cartilage. Within the hyaline cartilage are blood vessels that mark the beginnings of the secondary ossification enter. The primary ossification enter was in the midway down the bone shaft. This will be discussed later. The lacunae of the cartilage cells (chondrocytes) here are spaced far apart; the chondrocytes are not dividing. This zone is called the resting zone, which is appropriate enough. The cartilage will remain at the surface of the bone here to become the articulating cartilage of the joint. Now examine the zone inferior to the resting zone. You should just be entering your arch. Note that the chondrocytes are stacked together in tight columns. This is the zone of proliferation. The chondrocytes are dividing rapidly, making the bone grow longer. The zone just below the zone of proliferation shows fat chondrocytes, rich in glycogen. This is the zone of hypertrophication. The next zone, is the zone of calcification, right at the edge of the developing bone. Now the chondrocytes are dying, leaving behind a strips of calcified matrix known as spicules. The spicules will be invaded by osteoblasts and true bone will be made using the calcium as a framework. Below this zone is the zone of ossification, the region of new bone formation. Now examine these zones at 40x. You should be able to see chondrocytes clearly in the first three zones. You should be able to see osteoblasts clearly in the zone of calcification. Osteoblasts are also abundant in the trabeculae. The free cells not associated with the trabeculae are developing blood cells in red bone marrow. Try to find a large multinucleated cell. This cell is a megakaryocyte, which forms blood platelets. There is also a chance of finding an osteoclast. This cell will be more or less football shaped with three or more nuclei (see p. 81, figure 4). Osteoclasts break down bone to provide a space for bone reformation. Our skeletons are constantly being broken down and reformed in a process called remodeling.

Draw what you can see in endochondral bone formation