1. 1 Bone Blood & Hemopoiesis Jun Zhou ( 周俊 ) School of Medicine, Zhejiang University 20150324

2. 2  Bones are Organs of the Skeletal System Consist of: -bone tissue -other C.T. hemopoietic tissue fat tissue hyaline cartilage -blood vessels -nerves  Classified as: - compact (dense) bone ( 骨密质 ) -spongy (cancellous) bone ( 骨松质 )  Classified by shape: - Long, Short, Flat, and Irregular Bones

3. 3 BONE Osseous tissue: bone matrix +cells Bone matrix: Calcium phosphate forms: Hydroxyapatite crystals( 羟磷灰石 ), Ca 10 (PO 4 ) 6 (OH) 2 Type I and some type V collagen make up 90% of matrix- Ground substance- -glycosaminoglycans( 糖胺多糖 ) -glycoproteins -sialoproteins Cells : osteocytes-

4. 4 Bone matrix ( 骨质 ) --- organic matter:  bone collagen fiber -collagenous fiber (type I collagen)  ground substance: glycosaminoglycan,osteocalcin （骨钙蛋白）

5. 5 ---inorganic matter: bone salts Hydroxyapatite crystal:  Ca 10 (PO 4 ) 6 (OH) 2  pin-shaped  10-20 nm  longitudinal arranged *bone lamella ( 骨板 ): parallel collagen + bone salts

7. 7 Bone: -is a specialized connective tissue with a mineralized matrix (hydroxyapatite crystals,Calcium Phosphate). - cells of bone are: osteocytes: （骨细胞） bone cells osteoprogenitor cells: （骨原细胞） give rise to osteoblasts osteoblasts: （成骨细胞） developing osteocytes osteoclasts: （破骨细胞） bone resorbing cells

8. 8 A. Osteoprogenitor cell （骨原细胞） Structure:  in periosteum and endosteum  fusiform, small  ovoid nucleus  slight basophilic cytoplasm Function:  differentiated into osteoblast and chondrocyte

9. 9 B. Osteoblast （成骨细胞） --- structure: LM:  single layer of cuboidal or low columnar cell  round nucleus  basophilic cytoplasm  located on the surface of bone tissue

10. 10 EM:  fine processes  rich in RER, Golgi complex Function: Synthesize bone collagen fiber and ground substance- osteoid ( 类骨质 ) Regulate mineralization, ALPase (Alkaline phosphatase) Become osteocytes Calcitonin ( 降钙素 )

11. 11 C. Osteocyte （骨细胞） --- structure:  flattened cell with multiple long thin processes  located in bone lacuna and bone canaliculus  basophilic cytoplasm  adjacent cells connect in bone canaliculus by gap junctions （缝隙连接） ---function:  maintain bone matrix  regulate the balance of calcium （钙） and phosphonium （磷）

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13. 13 D. Osteoclast （破骨细胞） ---Structure: LM  multinuclear large cell, 30-100um  2-100 nuclei  acidophilic cytoplasm  located at peripheral part of bone

14. 14 EM:  ruffled border （皱褶缘） -processes  light zone: --under the ruffled border --microfilament （微丝）  lysosome, RER, etal. TRAP （抗酒石酸磷酸酶） ---Function: dissolve and absorb bone matrix

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16. TRAP 染色，有酒红色染色为 +

17. 17 Architecture of long bone Compact bone Spongy bone Periosteum and endosteum

19. 19 Compact Bone （骨密质） a. circumferential lamella: outer/inner b. Haversian system (osteon 骨单位 ) c. interstitial lamella （间骨板）

20. 20 Haversian system (osteon):  central canal: N, BV, CT  Haversian lamella: 4-20 layers

21. 21 Interstitial lamella （间骨板）  irregular lamella  remnant of Haversian or circumferential lamella *Perforating canal （穿通管） (Volkmann canal)  transverse canal  Connect with Haversian canal Cement line （黏合线）  more bone salts  less collagen

22. 22 Spongy bone （骨松质） Trabeculae （骨小梁）  parallelly-arranged lamella  spongy-liked network Bone marrow:  hemopoietic tissue

23. 23 Periosteum and endosteum Periosteum （骨外膜） : DCT  outer layer more fiber bundles form perforating fiber  inner layer BV, N and osteoprogenitor cells Endosteum （骨内膜） : thin, osteoprogenitor cell and CT Function  provide nutrition  osteoblast for bone growth and repairing

24. 24 Osteogenesis

25. 25 Formation : osteoprogenitor cell→ osteoblast → osteoid ↓ ↓calcification osteocyte + bone matrix bone tissue Absorption: osteoclast →dissolve bone tissue→reconstruction

26. 26 A. Intramembranous ossification （膜内成骨） --- CT membrane →osteoprogenitor cell → osteoblast→ossification center→bone trabeculae →thicker and longer ---flattened bone 、 irregular bone

27. 27 B. Endochondral ossification （软骨内成骨） e.g. long bone

28. 28 Blood & Hemopoiesis

29. 29 Overview of Blood A fluid CT, circulates through the cardiovascular system Consists of cells and extracellular component Total volume: 5-6L Amounts to 7-8% of total body weight Function: 1) Transport nutrients and oxygen to cells 2) Transport wastes and carbon dioxide away from cells 3) Delivery hormones and regulatory substances 4) Maintenance of homeostasis 5) Transport of humoral agents and cells of the immune system

30. 30 Blood Composition ---formed elements: 45%  Erythrocyte （红细胞）  Leukocyte （白细胞）  Platelets （血小板） ---plasma( 血浆 ): 55%, PH 7.2-7.4, 90% water  plasma protein (albumin, globulin, fibrinogen), lipidprotein, enzymes, hormone, vitamin, inorganic salt and products of metabolism *serum( 血清 ): is plasma in which the fibrinogen has been removed by clotting

31. 31 Blood smear: Wright or Giemsa stain methods

32. 32 1. Cells

33. 33 Erythrocyte, RBC( 红细胞 ) Normal number male: (4.0-5.0)×10 12 /L female:(3.5-4.5)×10 12 /L Structure biconcave discs 6.5-8.0 μm, 1-2μm Anucleate, no organelles hemoglobin, Hb( 血红蛋白 ) 120-150g/L in male 105-135g/L in female Function: combine and transfer O 2 and CO 2 Life span: 120 days

34. 34 RBC cytoskeleton maintain shape spectrin and actin 血影蛋白 SPECTRIN ACTIN 肌动蛋白

35. 35 Blood type antigen ABO blood type Surface antigen of A B membrane protein

36. 36 Hemolysis ( 溶血 ) erythrocyte ghost Anemia ( 贫血 ) RBC <3.0 × 10 12 /L or Hb <100g/L

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38. 38 * Reticulocyte ( 网织红细胞 ) 0.5-1% of total erythrocyte 3-6% in newborn baby immature cell remained ribosome-delicate network structure when stained with brilliant cresyl blue ( 亮甲酚蓝, 煌焦油蓝 )

39. 39 Leukocyte ( 白细胞 ) large cells with nucleus involve in defense and immune reaction normal number: (4.0-10.0) × 10 9 /L classification:  granulocytes: /neutrophil 50-70% /eosinophil 0.5-3% /basophil 0-1%  agranulocytes: /lympocyte 20-30% /monocyte 3-8%

40. 40 Neutrophil ( 中性粒细胞 ): Proportion: 50-70% Structure: LM:  round, Ф10-12um  rob-liked or poly morphous nucleus, 2-5 lobes  fine neutrophilic granules -pink cytoplasm

41. 41 EM: specific granule:  80%  small, 0.3-0.4 um, ovoid or irregular in shape  lysozyme( 溶菌酶 ), phagocytin( 吞噬素 )(defensin) azurophilic granule: ( 嗜天青颗粒 )  20%  large, round or ovoid, 0.6-0.7 um  acid phosphatase (ACP), peroxidase, etc.

42. 42 Function: phagocytose bacteria  specific g.-kill B  azurophilic g.- digest B Life span: 6-8 hours in blood, 2-3 days in CT

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44. 44 Eosinophil ( 嗜酸粒细胞 ) Proportion: 0.5-3% LM:  round, Ф 10-15 um  2 lobes nucleus  Uniform acidophilic granules

45. 45 EM: granules:  round or ovoid  with cube-liked electron dense crystal  contain: -ACPase -histaminase -peroxidase Function:  counteract the infection of parasite （寄生虫感染 ）  reduce allergic reaction （过敏反应） Life span: 6-8 hour in blood, 8-12 days in CT

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47. 47 Basophil ( 嗜碱粒细胞 ): Proportion: 0-1% LM:  less, round, Ф 10-12 um  Large irregular basophilic granules obscure the nucleus  Metachromatic ( 异染性 ),Heparin, Histamin Function: involve in allergic reaction

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49. 49 Monocyte ( 单核细胞 ) Proportion: 3-8% LM:  large round, Ф 12-18 um  nucleus appear as kidney, horse-shoe or ovoid in shape  cytoplasm: gray-blue in color, contain azurophilic g.

50. 50 EM: azurophilic granule Function: Actively mobile and chemotaxis Life span: 1-5 days in blood macrophage-mononuclear phagocytic system in CT

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52. 52 Lymphocyte ( 淋巴细胞 ) 20-30% LM: small 6-8 um medium-sized 9-12 um large 13-20 um round nucleus with indentation, chromatin dense cytoplasm: /basophilic:bright blue in color /less azurophilic granule

53. 53 Classification: T 75% cellular immune reaction, regulate immune response B 10-15% plasma cell, humoral immune response large granular cell: NK( nature killer cell), 10% Function: involve in immune response

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56. 56 Blood platelet ( 血小板 ): cytoplasmic fragment of megakaryocyte （巨核细胞） normal number: 100-300×10 9 /L structure: LM:  disc-liked, 2-4um → irregular in shape Stimulation  in groups

57. 57 EM: tubular systems: opening tubule system dense tubule system granules: specific granule dense granule Function: clotting and stopping the bleed protecting endothelium

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60. 60 Hemopoiesis

61. 61 Places for hemopoiesis Yolk sac (3W) （卵黄囊） Liver (6W) Spleen (3M) Bone marrow (4-5M)

62. 62 Structure of bone marrow: red bone marrow hemopoietic tissue:  reticular CT  Hemopoietic cell,macrophage,fibroblast, mesenchymal cell  sinusoid

63. 63 Hemopoietic inductive microenvironment ( HIM): BV, fiber, extracellular GS hemopoietic stromal cells Reticular cell Fibroblast Macrophage Endothelial cell Lipid-laden cell

64. 64  All blood cells come from a single stem cell, the pluripotential stem cell. It forms two kinds of daughter cells: A. The lymphoid multipotential cell migrates to lymphoid organs. B. The myeloid multipotential cell remains in the marrow and from it come various colony forming cells(erythrocyte, megakaryocyte, monocyte, and granulocyte)

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66. 66 Regulations:  size :smaller and smaller  Nucleus :smaller and smaller or disappears chromatin: from fine and loose to rough and dense  cytoplasm: -more and more -basophilic: weaker and weaker -specific granule: more and more  ability to division: from have to no, but keep the potentials

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68. 68  THE END!