CARTILAGE AND BONE Department of Histology and Embryology, Faculty of Medicine MU pvanhara@med.muni.cz Petr Vaňhara, PhD §CARTILAGE General features: §specialized connective tissue with continuous ECM §flexible, mechanically resistant §avascular, non-innervated §support of soft tissues - trachea, larynx §skeletal support - costal cartilages §diarthrosis - joints §bone growth http://cdn4.arthrex.com/taxon-images/foot_and_ankle_cartilage_allograft/foot_and_ankle_cartilage_al lograft_0-large.png 1.cells 2. 2.fibrils 3. 3.amorphous ground substance CARTILAGE §Perichondrium – connective tissue around cartilage (except joints) §Extracellular matrix – water, proteoglycans and collagen fibrils § § § § § § §Cells of cartilage - chondroblasts, chondrocytes Solid consistence Pressure elasticity Growth ECM production Nutrition Growth CARTILAGE – COMPOSITION AND STRUCTURE http://image.slidesharecdn.com/cartilagelecture-150319145858-conversion-gate01/95/cartilage-lecture -6-638.jpg?cb=1426777533 Hyaline Elastic Fibrous DISTRIBUTION 40-day Old Cat Fetus http://chickscope.beckman.uiuc.edu/explore/embryology/day14/dev2.html Hyaline Alcian Blue&Alizarin Red - most abundant - temporary embryonal/fetal skeleton - epiphyseal growth plate - articulation (joints) respiratory passages DISTRIBUTION •mesenchymal origin •typical ultrastructure of proteosynthetically active cells •production of extracellular matrix •interstitial proliferation •isogenetic groups, lacunae §Chondroblasts and chondrocytes CELLS OF CARTILAGE Runx2 Osx Sox9 Runx2 C/EBP PPARg Collagen 2A aggrecan DIFFERENTIATION OF CHONDROBLASTS Chondrocytes are mesenchymal cells, like fibroblasts, osteoblasts and adipocytes •Chondroblasts proliferate, acquire capability to produce huge amounts of ECM and differentiate to chondrocytes. •Chondrocytes are postmitotic, fully differentiated cells http://histonano.com/books/Junqueira's%20Basic%20Histology%20PDF%20WHOLE%20BOOK/New%20folder%207/lo adBinary.gif DIFFERENTIATION OF CHONDROBLASTS •oval ® round cells •rich in organelles, especially rER and GA •glycogen granules (anaerobic metabolism) •occasionally lipid droplets http://intranet.tdmu.edu.ua/data/kafedra/internal/histolog/classes_stud/en/med/lik/ptn/1/08%20Chond roid%20tissue.%20Bony%20tissue.%20Muscular%20tissues..files/image002.jpg ULTRASTRUCTURE OF CHONDROBLASTS ULTRASTRUCTURE OF CHONDROBLASTS ULTRASTRUCTURE OF CHONDROBLASTS §Extracelullar matrix biomechanical properties HOW IT WORKS? glycosaminoglycans proteoglycans fibers water linear unbranched polysaccharides containing a repeating disaccharide unit: 1. N-acetylgalactosamine (GalNAc) or N-acetylglucosamine (GlcNAc) 2. uronic acid (glucuronate (GlcA)) or iduronate. hyaluronic acid http://www.chemie-im-alltag.de/english/articles/0128/Hyaluronsaeure.png GLYCOSAMINOGLYCANS IN CARTILAGE Glycosaminoglycan Localization Hyaluronic acid Umbilical cord, synovial fluid, fluid of corpus vitreum, cartilage Chondroitinsulphate Cartilage, bone, cornea, skin, notochord, aorta Dermatansulphate Skin, ligaments, adventitia of aorta Heparansulphate Aorta, lungs, liver, basal membranes Keratansulphate Iris, cartilage, nucleus pulposus, anulus fibrosus GLYCOSAMINOGLYCANS IN CARTILAGE Chondroitinsulphate Keratansulphate •proteoglycans -protein + dominant linear saccharide component -proteoglycan aggregates -water-binding – 80%, volume dependent of hydratation -aggrecan (cartilage) -syndekan -fibroglykan PROTEOGLYCANS AND FIBERS Výsledek obrázku pro aggrecan •collagen fibrils -col II + col IX/XI -thin fibrils (15-20 nm ® no striation) that do not form fibers like col I -interconnected with perichondrium -elastic fibers •pericellular •territorial •interterritorial transduction of biochemical and biomechanical signals TISSUE ARCHITECTURE OF CARTILAGE ECM 22 •pressure elasticity -proteoglycans – polyanionic (COO-, SO4II-) -expansion prevented by collagen fibrils -repulsion forces •biphasic model of cartilage conditioned by ECM composition -proteoglycans, collagen, cells, and lipids constitute the solid phase of the mixture -interstitial fluid that is free to move through the matrix fluid phase) -under impact loads, fluid flows through the framework, until the cartilage start to behave as a single-phase, incompressible, elastic solid - the fluid does not flow -after load release, fluid returns -nutritive aspect TISSUE ARCHITECTURE OF CARTILAGE ECM I.tangential (superficial) zone II. II.transitional zone III. III. III. III.radial (deep) zone IV. IV. tide mark I.mineralized cartilage zone II. subchondral bone • •synovial cartilage TISSUE ARCHITECTURE OF CARTILAGE ECM 1-Hyaline%20Cartilage%20-%20Trachea Exchange of metabolites Apositional growth Interstitial proliferation NUTRITION AND GROWTH §stratum fibrosum -biomechanics §stratum chondrogenicum -growth PERICHONDRIUM §membrana fibrosa -dense collagen c.t. §membrana synovialis §intima, subintima -folds extending to the joint cavity -numerous blood and lymphatic vessels, nerves -discontinuous cell layers (synovialocytes) -basal membrane and intercellular junctions absent - not an epithelium: mesenchymal (c.t.) origin -synovial fluid rich in hyaluronans -bursae synoviales, vaginae tendineum http://www2.indstate.edu/thcme/mmmoga/histology/synov1.jpg http://www2.indstate.edu/thcme/mmmoga/histology/synov2.jpg http://www2.indstate.edu/thcme/mmmoga/histology/slide35.html http://www.mdpi.com/lubricants/lubricants-01-00102/article_deploy/html/images/lubricants-01-00102-g 003-1024.png SYNOVIUM https://www.dartmouth.edu/~anatomy/Histo/lab_2/bone/DMS090/53.gif https://www.dartmouth.edu/~anatomy/Histo/lab_2/bone/DMS090/53.gif -acidophilic elastic fibers dispersed in matrix -no isogenetic groups -auricula, meatus, larynx, epiglottis elastická chrupavka ELASTIC CARTILAGE •less abundant than collagen •polymer – tropoelastin •minimal tensile resistance, loss of elasticity if overstretched •reduction of hysteresis = allow return back to original state after mechanic change http://legacy.owensboro.kctcs.edu/gcaplan/anat/Histology/elastic.gif http://www.udel.edu/Biology/Wags/histopage/empage/ect/ect3.gif http://www.nature.com/srep/2013/130620/srep02043/images/srep02043-f1.jpg ELASTIC FIBERS ELASTIC CARTILAGE •fibrous compound dominant – collagen I and II – mechanical durability •minimum of amorphous matrix-fibers visible •intervertebral discs, symphysis pubis, articular discs, meniscus • http://www2.indstate.edu/thcme/mmmoga/histology/invert.jpg FIBROCARTILAGE •Cartilage – no innervation, no vascularization – no spontaneous regeneration •No migration of chondrocytes to site of damage •Initiation of other degenerative events leading to cartilage erosion (arthritis) • Therapy: •joint mobility •restoration of biochemical and biophysical parameters of cartilage •prevention of further damage •removal of damaged tissue, autologous transplantation, MSCs on biocompatible scaffolds http://images.iop.org/objects/med/news/10/4/48/pic1.jpg CLINICAL CORRELATION §BONE •Primary (woven, fibrous) -Temporary, growth and regeneration of bones, collagen fibrils woven -Replaced by secondary bone -Remains only in some parts of body - sutures of skull, tuberositas ossium, tooth cement •Secondary (lamellar) -Lamellae – collagen fibers in concentric layers (3-7mm) around a canal with capillaries = Haversian system (osteon) File:Transverse Section Of Bone.png HISTOLOGICAL CLASSIFICATION OF BONE TISSUE http://upload.wikimedia.org/wikipedia/commons/e/ec/Woven_bone_matrix.jpg PRIMARY (WOVEN) BONE -Temporary, growth and regeneration of bones, collagen fibrils woven -Replaced by secondary bone -Remains only in some parts of body - sutures of skull, tuberositas ossium, tooth cement http://upload.wikimedia.org/wikipedia/commons/e/ec/Woven_bone_matrix.jpg Výsledek obrázku pro primary bone SECONDARY (LAMELLAR) BONE Výsledek obrázku pro trabecular compact bone Bone-eng •Lamellae – collagen fibers in concentric layers (3-7mm) around a canal with capillaries = Haversian system (osteon) •Spongy (trabecular) -Trabeculae, similar to compact -Epiphyses of long bones, short bones, middle layer of flat bones of the skull (diploe) •Compact - Outer and inner coat lamellae typical Haversian systems - Volkmann’s canals - Interstitial canals https://www.dartmouth.edu/~anatomy/Histo/lab_2/bone/DMS090/54.gif •Outer surface -Synovial joint – hyaline cartilage -Periosteum (periost) – membrane – dense CT, inner layer (osteoblasts) and outer layer (fibrous CT) -Inactive bone - fibrous CT in periost dominant -Collagen fibers – parallel to the bone surface -Sharpey’s fibers fix periost to the bone BONE SURFACES http://farm7.static.flickr.com/6140/6003152325_a91c2a820e_b.jpg National Museum of Natural History NY, USA BONE SURFACES http://www2.indstate.edu/thcme/mmmoga/histology/musinbo.jpg BONE SURFACES Inactive Active https://upload.wikimedia.org/wikipedia/commons/5/5a/619_Red_and_Yellow_Bone_Marrow.jpg •Inner surface – lining of cavities -medullar cavity -endosteum (endost) – single cell lining – bone remodeling -red bone marrow – hematopoiesis -yellow and gray bone marrow – adipocytes or CT -rich vascularization -hematopoietic niche File:Gray72-en.svg http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab8/IMAGES/BONE%20BLOOD%20SUPPLY.jpg BONE SURFACES bone_marrow_40X ENDOSTEAL SURFACE OF COMPACT BONE -60% mineral compound, 24% organic compound 12% H20, 4% fat -crystals – calcium phosphate, hydroxyapatite http://bio2tech.com/resources/generic+sem+bone.jpg BONE MATRIX http://www.nature.com/nature/journal/v412/n6846/images/412491aa.2.jpg BONE MATRIX -lining bone surface -produce ECM – collagen (I) and noncollagenous proteoglycans, glycoproteins -basophilic cytoplasm, rER, well developer Golgi Apparatus -euchromatin nucleus - -osteocytes embedded in matrix -canalliculi ossium Soubor:Active osteoblasts.jpg CELLS OF BONE – OSTEOBLASTS http://www.mhhe.com/biosci/ap/histology_mh/osteocyt.jpg http://www.gla.ac.uk/ibls/US/fab/images/generic/boblast.gif SEM of osteocytes Soubor:Osteocyte 2.jpg CELLS OF BONE – OSTEOCYTES -multinuclear, formed by fusion of mononuclear macrophages -bone matrix resorption http://www.technion.ac.il/~mdcourse/274203/slides/Skeletal%20Tissues/6-Osteoclasts%20-%20Intramembr anous%20Ossification.jpg http://bp0.blogger.com/_AOuMhsgQteY/RlyXaYXhvLI/AAAAAAAAADo/_xdySV3WRMs/s320/boyde_bfast.jpg CELLS OF BONE – OSTEOCLASTS http://www.intechopen.com/source/html/18309/media/image4.jpg CELLS OF BONE – OSTEOCLASTS File:Osteoclast1.jpg -complex architecture -enzymes degrading organic matrix -HCl TEM of Osteoclast CELLS OF BONE – OSTEOCLASTS •Intramembraneous •Endochondral http://www.nature.com/labinvest/journal/v96/n2/images/labinvest2015142f1.jpg http://img.medscape.com/pi/features/slideshow-slide/osteoporosis/fig5.jpg BONE OSSIFICATION INTRAMEMBRANEOUS OSSIFICATION 2 1 3 4 5 ENDOCHONDRAL OSSIFICATION http://www.nature.com/nrendo/journal/v11/n12/images/nrendo.2015.165-f2.jpg ENDOCHONDRAL OSSIFICATION http://d2qiws50qrj9uc.cloudfront.net/content/develop/142/5/817/F2.large.jpg ENDOCHONDRAL OSSIFICATION https://upload.wikimedia.org/wikipedia/commons/1/10/Proximal_tibia_Masson_Goldner_Trikrom_rabbit_60 0x_growth_zone.jpg GROWTH PLATE GROWTH GROWTH PLATE GROWTH http://ns.umich.edu/Releases/2005/Feb05/img/bone.jpg http://ns.umich.edu/Releases/2005/Feb05/img/bone.jpg BONE REMODELLING http://www.nature.com/nrrheum/journal/v11/n1/images/nrrheum.2014.164-f2.jpg http://library.med.utah.edu/WebPath/jpeg3/BONE015.jpg 1. Reactive phase - fracture and inflammatory phase - granulation tissue formation Reparative phase - cartilage callus formation - lamellar bone deposition Remodeling phase - remodeling to original bone shape 1. CLINICAL CORRELATIONS – FRACTURE HEALING •OSTEOPOROSIS http://1.bp.blogspot.com/-RlmeWVPsF9w/TangAgX0qFI/AAAAAAAAAHg/MiiJLkYvMZ8/s1600/osteoporotic+bone.j pg • OSTEOPETROSIS http://images.radiopaedia.org/images/130505/24e8e4473dafe5ee65962c39037049_big_gallery.jpg • REVMATOID ARTHRITIS http://www.rheumatoidarthritistreatment101.com/wp-content/uploads/2011/04/1MAY.jpg CLINICAL CORRELATIONS – DISBALANCE OF BONE HOMEOSTASIS •PAGET DISEASE Výsledek obrázku pro PAGET disease Bone Loss in Space: “Bag of Bones” DISBALANCE OF BONE HOMEOSTASIS Meet the real ironman of spaceflight: Valery Polyakov | Ars Technica Valerij Poljakov 438 days in sapce MIR ? Synarthrosis -joint by intercalated tissue (catilage, bone or c.t.) -Synostoses – joint by bone tissue – os coxae, os sacrum -Synchondrosis – joint by hyaline cartialge – development of synostosis -Symphysis – joint by fibrocartilage– os pubis, intervertebral discs -Syndesmosis – dense collage regular c.t. – sutures of skull, gomphosis - Diarthrosis -synovial joint -hyaline cartilage without perichondrium -cartialge calcification in site of attachment to the bone -joint capsule -Stratum fibrosum -Stratum synoviale - meniscus – fibrocartialge, avascular, without inervation - tendons – dense collagen regular c.t., elastic fibers - bursae – like joint capsule mc_ch08_fig01 Image581 JOINTS http://www.med.muni.cz/histology http://ecx.images-amazon.com/images/I/41RNW9VE8OL._SX359_BO1,204,203,200_.jpg Guide to General Histology and Microscopy Anatomy FURTHER STUDY https://s-media-cache-ak0.pinimg.com/736x/80/3c/6a/803c6afaa0f98409c9117ff64d69de69.jpg Thank you for attention