LECTURER: Dennis Ivan U. Bravo, MD DATE: 15 July 2014 FEU-NRMF Institute of Medicine Batch 2018 Outline: Review of bone and BM histology Erythropoiesis Granulopoeisis Platelet formation Clinical Correlation
REVIEW OF BONE
FUNCTION OF BONE Support for soft tissues and the attachment site for tendons. Because the bone is very hard, It protects the soft internal organs like your brain. It is protected from the external environment by the very hard skull. In combination with the muscle, it is responsible for locomotion. The bone also has a biochemical function. It stores mineral (e.g. calcium and phosphates) and is released whenever theres a need for them. Blood cell production. It also has another biological function, which is to produce the cells that are found in the blood. Triglyceride storage. It stores triglyceride in the form of yellow marrow.
Explanation: This is a cross-section of a bone showing you two types. The one in the surface is the CORTICAL (Compact) BONE. So if its enlarged, you can see that its made up of units known as OSTEON and in the center, you will find the HAVERSIAN CANAL and the ones concentrically arranged around it are the OSTECYTES. Pointed above is the TRABECULAR (Spongy) BONE. The trabecular bone itself is not in the picture but its actually the marrow which fills the spaces within the trabecular bone.
HISTOLOGY OF BONE MARROW
TYPES OF BONE MARROW (OVERVIEW) Red Marrow Actively involved in hematopoeisis. o Contents: Stem Cells, Progenitor Cells, Developing Erythroblasts, Myeloblasts, and Megakaryoblasts. o Highly cellular. The spaces are filled with actively dividing hematopoietic cells. Yellow Marrow storage site for adipocytes. o Contents: Adipose cells (varies with age and activity) Note: Both marrows contain a lot of dilated capillaries. Theres a wide network of capillaries present in the marrow.
YELLOW MARROW Located on long bones of adults. (e.g. femur) Highly infiltrated with fat. Not hematopoietic under normal conditions. But it has a potential to become when theres a need for extra hematopoietic cell. 1 | P a g e M I C R O H S B A 2.1 MYELOID TISSUE HISTOLOYGY LECTURER: Dennis Ivan U. Bravo, MD DATE: 15 July 2014 FEU-NRMF Institute of Medicine Batch 2018
Explanation: This is how it looks like under a microscope. The yellow marrow is composed of spaces between trabecular bones and is filled with several adipose cells. The adipose cells are filled with triglycerides.
RED MARROW Located in the epiphysis of long bones, flat, irregular, and short bones. (e.g. Sternum) Highly vascular with large dilated venous sinuses and surrounding them are the active hematopoietic cells.
Explanation: The red marrow is filled with several venous sinuses, arising from an artery. The green-colored objects are the RETICULAR FIBERS, which provides support to the hematopoietic cells.
Explanation: The clusters of solid blue cells are the ERYTHROPOIETIC CELLS. The clusters of granulated blue cells are the GRANULOPOIETIC CELLS. The white space filled with RBCs is the VENOUS SINUS. It is normal to find adipose cells. The distribution for adults is 50% hematopoietic cells and the remaining 50% are adipose cells.
Explanation: There is a prominent VENOUS SINUS filled with RBCs. Surrounding them will be the HEMATOPOIETIC CELLS, specifically ERYTHROPOIETIC CELLS. A large cell is also present here known as the MEGAKARYOCYTE.
Amount of red marrow varies according to the patients age. o Childhood: Virtually present in all bones of the body. It is composed of hematopoietic cells. Virtually no adipose cells are found. o Adults: Reduced to 50% Present in the sternum, ribs, pelvis and skull. o 70 years: Reduced to 30%
2 | P a g e M I C R O H S B A 2.1 MYELOID TISSUE HISTOLOYGY LECTURER: Dennis Ivan U. Bravo, MD DATE: 15 July 2014 FEU-NRMF Institute of Medicine Batch 2018
HOW DO WE DISTINGUISH ERYTHROPOIETIC CELLS?
FROM OTHER HEMATOPOIETIC CELLS o As a group, they do not have any prominent granules in contrast to the developing myelocytes. o The nucleus, whether small or large, is consistently round when its still present. In contrast to the developing WBCs where the shape of the nucleus varies according to stage of its development.
FROM EACH STAGE o Cell size o Subtle differences in the nucleus o Color of the cytoplasm
TREND IN THE DEVELOPMENT OF RBCs
Explanation: If the cell develops from young to mature, youll notice a change in the nucleus. Initially, it is big it becomes smaller until eventually disappears. There is a PROGRESSIVE CONDENSATION of the nucleus. Another trend is that there is a change from Basophilic to Eosinophilic. Initially, there is no haemoglobin. But eventually, an accumulation of haemoglobin will be present as the cell progress thereby giving it its eosinophilic quality. Since there is absence of haemoglobin during the early stage of erythropoiesis, there is a production of RNA present. These RNA will contribute to the production of RIBOSOMES. This gives the cell its basophilic characteristics. The RNA and ribosome are the ones involved in protein synthesis, namely haemoglobin. There is also a progressive reduction in the size of the cell. It initially starts with a large cell and as it matures, the cell is now smaller.
STAGES OF ERYTHROPOIESIS
BLAST CELL o The origin of erythropoietic cells. o Cannot be distinguished from the blast cells of myelocytes series. o Mitotically active (It divides). o No granules. PROERYTHROBLAST o Cell is large. o Mitotic. o Positive nucleoli (w/c means that its actively involved in synthesis of RNA). o Basophilic cytoplasm. BASOPHILIC ERYTHROBLAST o The nucleolus has disappeared (w/c means the synthesis of RNA has stopped). o Intensely basophilic because of ribosomes. POLYCHROMATIC ERYTHROBLAST o Smaller than its precursor. o The nucleus is starting to condense thats why it has a checkerboard appearance (Writes stain). o Gray-green cytoplasm because of haemoglobin accumulation. ORTHOCHROMATIC ERYTHROBLAST o Aka Normoblast o Smaller pyknotic nucleus (about to be extruded) o Pink cytoplasm due to haemoglobin accumulation. RETICULOCYTE o No nucleus (already extruded) o Pink/orange color (because of the haemoglobin) with bluish hue (due to remaining ribosomes). o Named such because of cresyl blue. They seem to have a network-like appearance.
3 | P a g e M I C R O H S B A 2.1 MYELOID TISSUE HISTOLOYGY LECTURER: Dennis Ivan U. Bravo, MD DATE: 15 July 2014 FEU-NRMF Institute of Medicine Batch 2018
IN SUMMARY: Decrease in basophilia Increase in haemoglobin concentration Decrease in cell volume Increase in chromatin condensation, followed by extrusion of a pyknotic nucleus
GRANULOPOIESIS
All granulocytes arise from a single MYELOBLAST which came from a UNIPOTENT STEM CELL. The development of all types of myelocytes follows the same stages.
HOW DO WE DISTINGUISH GRANULOPOIETIC CELLS?
FROM OTHER HEMATOPOIETIC CELLS o Consistently contain granules o When clustered as a group, they appear DIRTY together.
FROM EACH STAGES o Type of granule present o Appearance of the nucleus
STAGES OF GRANULOPOIESIS
PROMYELOCYTE o First identifiable WBC precursor. o Contains purple-staining azurophilic nonspecific granules. o Cell is large. o Nucleus is large. o Nucleolus is prominent.
MYELOCYTE o Distinguished once the lilac-staining specific granules start appearing over the purple-staining azurophilic nonspecific granules. o Wide range in cell size. o Frequently indented round nucleus.
METAMYELOCYTE o The primary granules will start to disappear and the secondary granules will predominate. o Nucleus becomes flattened.
BAND o The nucleus will become more condensed. Eventually, it will become U-shaped or horseshoe-shaped.
GRANULOCYTE o The nucleus becomes segmented into lobes.
IN SUMMARY:
4 | P a g e M I C R O H S B A 2.1 MYELOID TISSUE HISTOLOYGY LECTURER: Dennis Ivan U. Bravo, MD DATE: 15 July 2014 FEU-NRMF Institute of Medicine Batch 2018
Myeloblast = No cytoplasmic granules Promyelocyte = appearance of Primary Granules Myelocyte = appearance of Secondary granules Metamyelocyte = Abundance of Secondary granules and dispersed Primary Granules Band = Nucleus will become horseshoe-shaped Granulocyte = Nucleus is segmented
PLATELET FORMATION
OVERVIEW: Consist of 3 stages and it arose from the Unipotent Stem Cell. It will divide to form the MEGAKARYOBLAST. The megakaryoblast will undergo ENDOMITOSIS to form MEGAKARYOCYTE. And the megakaryocyte will fragment to form the platelet.
STAGES OF PLATELET FORMATION
MEGAKARYOBLAST o Large cell with single large-lobed nucleus. o Basophilic non-granular cytoplasm. o It will undergo ENDOMITOSIS (nucleus is doubled up but doesnt undergo division). o RARE
MEGAKARYOCYTE o Extremely large cell with single, large POLYPLOID nucleus. o Lies just outside the sinusoids. o Fragments to platelets. o Sheds platelets. o They are found near the sinusoids because they form PSEUDOPODS that fragments to form platelets goes directly to the sinusoids to join the bloodstream or circulation right away.
CLINICAL CORRELATION
ACUTE MYELOGENOUS LEUKEMIA
LEUKEMIA the uncontrolled proliferation of the myelocyte. ACUTE MYELOGENOUS LEUKEMIA the uncontrolled proliferation of UNDIFFERENTIATED MYELOBLAST.
COMPARISON: Bone Marrow w/ AML o Highly cellular o Everythings obliterated o Absence of erythropoietic cells o Abundance of myeloblastic cells o Absence of fats o Absence of platelet forming cells o Obliteration of marrow space with tumor cells SYMPTOMS (Purely based on the slides): Increased WBC o Caused by the abundance in myeloblast but are immature and non-functional. Infection o WBCs are non-functional Anemia o Obliteration of erythropoietic cells Bleeding o Obliteration of platelet forming cells Pain o Since it is growing in the confined space of the bone
5 | P a g e M I C R O H S B A 2.1 MYELOID TISSUE HISTOLOYGY LECTURER: Dennis Ivan U. Bravo, MD DATE: 15 July 2014 FEU-NRMF Institute of Medicine Batch 2018