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Bone
Blood & Hemopoiesis
Jun Zhou (周俊)
School of Medicine, Zhejiang University
20140311
1
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
2
BONE
Osseous tissue: bone matrix +cells
Bone matrix:
Calcium phosphate forms:
Hydroxyapatite crystals(羟磷灰石), Ca10(PO4)6(OH)2
Type I and some type V collagen make
up 90% of matrixGround substance-glycosaminoglycans(糖胺多糖)
-glycoproteins
-sialoproteins
Cells : osteocytes3
Bone matrix (骨质)
---organic matter:
bone collagen fiber -collagenous fiber (type I
collagen)
 ground substance:
glycosaminoglycan,osteocalcin(骨钙蛋白)

4
---inorganic matter: bone salts
Hydroxyapatite crystal:




Ca10(PO4)6(OH)2
pin-shaped
10-20 nm
longitudinal arranged
*bone lamella (骨板):
parallel collagen + bone salts
5
骨
组
织
的
各
种
细
胞
和
骨
板
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
7
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
8
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
9
EM:


fine processes
rich in RER, Golgi complex
Function:
Synthesize
bone collagen fiber and ground substanceosteoid (类骨质)
Regulate mineralization, ALPase (Alkaline phosphatase)
Become osteocytes
Calcitonin (降钙素)
10
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 )
phosphonium(磷)
and
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D. Osteoclast
(破骨细胞)
---Structure:
LM

multinuclear large cell, 30-100um

2-100 nuclei

acidophilic cytoplasm

located at peripheral part of bone
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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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15
TRAP染色,有酒红色染色为+
Architecture of long bone



Compact bone
Spongy bone
Periosteum and endosteum
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Compact Bone(骨密质)
a. circumferential lamella: outer/inner
b. Haversian system (osteon 骨单位)
c. interstitial lamella (间骨板)
19
Haversian system (osteon):

central canal: N, BV, CT

Haversian lamella: 4-20 layers
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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
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Spongy bone(骨松质)
Trabeculae(骨小梁)
 parallelly-arranged lamella
 spongy-liked network
Bone marrow:
 hemopoietic tissue
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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
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Osteogenesis
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Formation:
osteoprogenitor cell→ osteoblast → osteoid
↓
↓calcification
osteocyte + bone matrix
bone tissue
Absorption:
osteoclast →dissolve bone tissue→reconstruction
25
A. Intramembranous ossification(膜内成骨)
---CT membrane →osteoprogenitor cell →
osteoblast→ossification center→bone trabeculae
→thicker and longer
---flattened bone、irregular bone
26
B. Endochondral ossification(软骨内成骨)
e.g. long bone
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Blood & Hemopoiesis
28
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

29
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
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Blood smear: Wright or Giemsa stain methods
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1. Cells
32
Erythrocyte, RBC(红细胞)
Normal number
male: (4.0-5.0)×1012/L
female:(3.5-4.5)×1012/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 O2 and CO2
 Life span: 120 days




33
RBC cytoskeleton
 maintain shape
 spectrin and actin
血影蛋白
SPECTRIN
肌动蛋白
ACTIN
34
Blood type antigen



ABO blood type
Surface antigen of A B
membrane protein
35
 Hemolysis (溶血)
erythrocyte ghost
 Anemia (贫血)
RBC <3.0×1012 /L
or
Hb <100g/L
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37
* 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)
38
Leukocyte (白细胞)
large cells with nucleus
 involve in defense and immune reaction
 normal number: (4.0-10.0) ×109/L
 classification:
 granulocytes: /neutrophil 50-70%
/eosinophil 0.5-3%
/basophil
0-1%
 agranulocytes:
/lympocyte 20-30%
/monocyte 3-8%

39
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
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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.
41
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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Eosinophil (嗜酸粒细胞)
Proportion: 0.5-3%
LM:
 round, Ф 10-15 um
 2 lobes nucleus
 Uniform acidophilic granules
44
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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Basophil (嗜碱粒细胞):
Proportion: 0-1%
LM:
round, Ф 10-12 um
 Large irregular basophilic granules obscure
the nucleus
 less,

Metachromatic (异染性),Heparin, Histamin
Function: involve in allergic reaction
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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.
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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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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
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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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Blood platelet (血小板):



cytoplasmic fragment of megakaryocyte (巨核细胞)
normal number: 100-300×109/L
structure:
LM:


disc-liked, 2-4um → irregular in shape
Stimulation
in groups
56
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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Hemopoiesis
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Places for hemopoiesis




Yolk sac (3W)(卵黄囊)
Liver (6W)
Spleen (3M)
Bone marrow (4-5M)
61
Structure of bone marrow: red bone marrow
hemopoietic tissue:
 reticular CT
 Hemopoietic cell,macrophage,fibroblast, mesenchymal cell
 sinusoid
62
Hemopoietic inductive microenvironment
( HIM):
 BV, fiber, extracellular GS
 hemopoietic stromal cells





Reticular cell
Fibroblast
Macrophage
Endothelial cell
Lipid-laden cell
63
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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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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THE
END!
68
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