title: Immobilization of naringin onto chitosan substrates by using ozone activation.
abstract: Ozone oxidation can easily produce peroxides containing active free radicals that can be used for the surface modification of biomaterials. This process is highly efficient and nontoxic. In this research, naringin, an HMG-CoA reductase inhibitor that can promote bone formation, was immobilized onto a chitosan film using ozone activation. First, a chitosan film was treated by ozone to produce peroxides; these peroxides were then quantified and their amount was optimized by an iodide assay. For the in vitro delivery of naringin, a chitosan-naringin substrate was immersed in phosphate-buffered saline to quantify the released amount of naringin. It was found that the immobilized naringin was slowly released over the course of two weeks, where its concentration in the medium was controlled by this delivery process. The results of cell culture showed that cell viability and early osteogenic differentiation, as measured by alkaline phosphatase expression, were promoted with the immobilized naringin on chitosan substrates. The expression of osteogenic proteins, including type-I collagen, bone siloprotein, and osteocalcin, were also enhanced. According to the results of Smad1 and Smad6 phosphorylation, immobilized naringin on ozonated chitosan substrates would be able to initiate bone morphogenetic protein-Smad signaling by activating receptor Smad and by suppressing inhibitory Smad. The results in this research demonstrated that the naringin-chitosan substrate produced by biocompatible ozone activation was highly osteoconductive without cytotoxicity.
authors: Division of Orthodontics & Pediatric Dentistry, Dental Department, Tri-Service General Hospital, Taipei, Taiwan.
authors: Chung Hsing
authors: CH
authors: Li
authors: Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10617, Taiwan.
authors: Jing Wei
authors: JW
authors: Wang
authors: Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10617, Taiwan. Electronic address: mhho@mail.ntust.edu.tw.
authors: Ming Hua
authors: MH
authors: Ho
authors: Division of Oral and Maxillofacial Surgeons, Dental Department, Tri-Service General Hospital, Taipei, Taiwan.
authors: Jia Lin
authors: JL
authors: Shih
authors: R&D Center for Membrane Technology, Chung Yuan University, Chungli, Taiwan.
authors: Sheng Wen
authors: SW
authors: Hsiao
authors: Department of Chemical Engineering, Can Tho University, Can Tho City, Viet Nam.
authors: Doan Van Hong
authors: DV
authors: Thien
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D024042:Collagen Type I / Q000378:metabolism; D044950:Flavanones / Q000494:pharmacology*; D058574:Integrin-Binding Sialoprotein / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D010126:Ozone / Q000494:pharmacology*; D010545:Peroxides / Q000032:analysis; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D051785:Smad Proteins / Q000378:metabolism; D013055:Spectrophotometry, Infrared; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D024042:Collagen Type I; D044950:Flavanones; D058574:Integrin-Binding Sialoprotein; D010545:Peroxides; D051785:Smad Proteins; D015675:Osteocalcin; D010126:Ozone; D048271:Chitosan; D000469:Alkaline Phosphatase; C005274:naringin
keywords: Chitosan; Naringin; Osteoconduction; Ozone; Surface modification