Cindy C Shu1, James Melrose2,3,4. 1. Raymond Purves Bone and Joint Research Laboratories, Level 10 Kolling Institute of Medical Research (B6), North Sydney Area Health Authority, University of Sydney at the Royal North Shore Hospital, St. Leonards, NSW, 2065, Australia. 2. Raymond Purves Bone and Joint Research Laboratories, Level 10 Kolling Institute of Medical Research (B6), North Sydney Area Health Authority, University of Sydney at the Royal North Shore Hospital, St. Leonards, NSW, 2065, Australia. james.melrose@sydney.edu.au. 3. Department of Surgery, Northern Clinical School, University of Sydney, Sydney, Australia. james.melrose@sydney.edu.au. 4. Graduate School of Biomedical Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia. james.melrose@sydney.edu.au.
Abstract
PURPOSE: The present study was designed to ascertain how altered biomechanics in adolescent idiopathic scoliotic (AIS) intervertebral discs (IVDs) affected tissue compositions and aggrecan processing compared to age matched and aged human IVDs. Newborn, 2- and 10-year-old ovine IVDs were also examined. METHODS: Aggrecan populations were separated by Sepharose CL2B chromatography, composite agarose polyacrylamide gel electrophoresis (CAPAGE) and identified by immunoblotting. The KS and CS content of IVD tissue extracts from AIS IVDs were compared with age-matched normal adolescent IVDs and with old human IVDs. Extracts from newborn, 2- and 10-year-old ovine IVDs were also examined in a similar manner. RESULTS: Adolescent idiopathic scoliotic IVD Aggrecan populations shared similar levels of polydispersity and aggregatability with hyaluronan as old IVD proteoglycans. CAPAGE demonstrated three aggrecan populations in AIS, aged human and ovine IVDs increased polydispersity and mobility in CAPAGE. AIS IVDs had GAG compositions similar to aged human and ovine IVDs. Sulphated KS (5-D-4) and chondroitin-6-sulphate, 3-B-3(+) were markers of tissue maturation, and chondroitin-4-sulphate, 2-B-6(+) was prominent in immature IVDs but its levels were lower in mature IVDs. DISCUSSION: Sulphated KS and 3-B-3(+) CS were prominently associated with IVD maturation and AIS IVDs, while the 2-B-6(+) CS isomer was associated with immature IVD tissues. The polydispersity of aggrecan in AIS IVDs, which was similar to in old human and ovine IVDs, reflected altered processing in the AIS IVDs in response to the biomechanical microenvironments the disc cells were exposed to in AIS IVDs. These slides can be retrieved under Electronic Supplementary Material.
PURPOSE: The present study was designed to ascertain how altered biomechanics in adolescent idiopathic scoliotic (AIS) intervertebral discs (IVDs) affected tissue compositions and aggrecan processing compared to age matched and aged human IVDs. Newborn, 2- and 10-year-old ovine IVDs were also examined. METHODS: Aggrecan populations were separated by Sepharose CL2B chromatography, composite agarosepolyacrylamide gel electrophoresis (CAPAGE) and identified by immunoblotting. The KS and CS content of IVD tissue extracts from AIS IVDs were compared with age-matched normal adolescent IVDs and with old human IVDs. Extracts from newborn, 2- and 10-year-old ovine IVDs were also examined in a similar manner. RESULTS: Adolescent idiopathic scoliotic IVD Aggrecan populations shared similar levels of polydispersity and aggregatability with hyaluronan as old IVD proteoglycans. CAPAGE demonstrated three aggrecan populations in AIS, aged human and ovine IVDs increased polydispersity and mobility in CAPAGE. AIS IVDs had GAG compositions similar to aged human and ovine IVDs. SulphatedKS (5-D-4) and chondroitin-6-sulphate, 3-B-3(+) were markers of tissue maturation, and chondroitin-4-sulphate, 2-B-6(+) was prominent in immature IVDs but its levels were lower in mature IVDs. DISCUSSION: SulphatedKS and 3-B-3(+) CS were prominently associated with IVD maturation and AIS IVDs, while the 2-B-6(+) CS isomer was associated with immature IVD tissues. The polydispersity of aggrecan in AIS IVDs, which was similar to in old human and ovine IVDs, reflected altered processing in the AIS IVDs in response to the biomechanical microenvironments the disc cells were exposed to in AIS IVDs. These slides can be retrieved under Electronic Supplementary Material.
Authors: Peter J Roughley; Lee I Melching; Terrence F Heathfield; Richard H Pearce; John S Mort Journal: Eur Spine J Date: 2006-05-31 Impact factor: 3.134
Authors: I A Nieduszynski; T N Huckerby; J M Dickenson; G M Brown; G H Tai; H G Morris; S Eady Journal: Biochem J Date: 1990-10-01 Impact factor: 3.857
Authors: Sharon Brown; James Melrose; Bruce Caterson; Peter Roughley; Stephen M Eisenstein; Sally Roberts Journal: Eur Spine J Date: 2012-02-23 Impact factor: 3.134
Authors: Mauro Alini; Stephen M Eisenstein; Keita Ito; Christopher Little; A Annette Kettler; Koichi Masuda; James Melrose; Jim Ralphs; Ian Stokes; Hans Joachim Wilke Journal: Eur Spine J Date: 2007-07-14 Impact factor: 3.134
Authors: Zhaoyang Liu; Amro A Hussien; Yunjia Wang; Terry Heckmann; Roberto Gonzalez; Courtney M Karner; Jess G Snedeker; Ryan S Gray Journal: Elife Date: 2021-07-28 Impact factor: 8.140
Authors: Naomi N Lee; Elias Salzer; Frances C Bach; Andres F Bonilla; James L Cook; Zulma Gazit; Sibylle Grad; Keita Ito; Lachlan J Smith; Andrea Vernengo; Hans-Joachim Wilke; Julie B Engiles; Marianna A Tryfonidou Journal: JOR Spine Date: 2021-06-14