title: Development of kartogenin-conjugated chitosan-hyaluronic acid hydrogel for nucleus pulposus regeneration.
abstract: Injectable constructs for in vivo gelation have many advantages in the regeneration of degenerated nucleus pulposus. In this study, an injectable hydrogel consisting of chitosan (CS) and hyaluronic acid (HA) crosslinked with glycerol phosphate (GP) at different proportions (CS : GP : HA, 6 : 3 : 1, 5 : 3 : 2, 4 : 3 : 3, 3 : 3 : 4, 2 : 3 : 5, 1 : 3 : 6, V : V : V) was developed and employed as a delivery system for kartogenin (KGN), a biocompound that can activate chondrocytes. In vitro gelation time, morphologies, swelling, weight loss, compressive modulus and cumulative release of KGN in hydrogels were studied. For biocompatibility assessments, human adipose-derived stem cells (ADSCs) were encapsulated in these hydrogels. The effects of KGN on stem cell proliferation and differentiation into nucleus pulposus-like cells were examined. The hydrogels with higher concentrations of HA showed a slightly shorter gelation time, higher water uptake, faster weight loss and faster KGN release compared to the hydrogels with lower concentrations of HA. As the KGN-conjugated hydrogel prepared with the proportions 5 : 3 : 2 displayed good mechanical properties, it was chosen as the optimal gel to promote cell proliferation and differentiation. No significant difference was seen in the expression levels of nucleus pulposus markers induced by KGN or TGF-β. Additionally, inclusion of KGN and TGF-β together did not produce a synergistic effect in inducing nucleus pulposus properties. In conclusion, we have developed a KGN-conjugated CS/HA hydrogel (5 : 3 : 2) with sustained release of KGN in hydrogel that can promote ADSC proliferation and nucleus pulposus differentiation. This kind of hydrogel may be a simple and effective candidate for the repair of degenerative NP tissue after minimally invasive surgery.
authors.affiliation: Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, China. yanxiazhu@szu.edu.cn.
authors.forename: Yanxia
authors.identifier: http://orcid.org/0000-0003-3247-3593
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Department of Spinal Surgery, Shenzhen Sixth People's Hospital (Nanshan Hospital), Shenzhen, 518060, China. szyiwh@163.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Tan
authors.affiliation: Department of Spinal Surgery, Xiaogan Maternity&Child Healthcare Hospital, Xiaogan, 432100, China.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, China. yanxiazhu@szu.edu.cn.
authors.forename: Guangyao
authors.initials: G
authors.lastname: Lin
authors.affiliation: Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, China. yanxiazhu@szu.edu.cn.
authors.forename: Fei
authors.initials: F
authors.lastname: Yin
authors.affiliation: Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, China. yanxiazhu@szu.edu.cn.
authors.forename: Liang
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China.
authors.forename: Kedong
authors.initials: K
authors.lastname: Song
authors.affiliation: Burns Research Group, ANZAC Research Institute, University of Sydney, Concord, NSW 2139, Australia.
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, China. yanxiazhu@szu.edu.cn.
authors.forename: Guangqian
authors.initials: G
authors.lastname: Zhou
authors.affiliation: Department of Spinal Surgery, Shenzhen Sixth People's Hospital (Nanshan Hospital), Shenzhen, 518060, China. szyiwh@163.com.
authors.forename: Weihong
authors.initials: W
authors.lastname: Yi
mesh_terms: D000813:Anilides / Q000008:administration & dosage* / Q000494:pharmacology; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D048271:Chitosan / Q000031:analogs & derivatives*; D003692:Delayed-Action Preparations / Q000737:chemistry*; D006801:Humans; D006820:Hyaluronic Acid / Q000031:analogs & derivatives*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D000070614:Nucleus Pulposus / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010795:Phthalic Acids / Q000008:administration & dosage* / Q000494:pharmacology; D017207:Rats, Sprague-Dawley; D012038:Regeneration / Q000187:drug effects*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*
chemical_list: D000813:Anilides; D003692:Delayed-Action Preparations; D010795:Phthalic Acids; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D006820:Hyaluronic Acid; D048271:Chitosan; C572342:kartogenin