title: The effect of calcium ion concentration on osteoblast viability, proliferation and differentiation in monolayer and 3D culture.
abstract: Our research group aims to develop an osteochondral composite using type II collagen gel with hydroxyapatite (HAp) deposited on one side. Soaking gels in Ca2+ and phosphate solution is indispensable to HAp deposition, so relationships between cell behavior and Ca2+ concentration were examined in two- and three-dimensional cultures. The present results indicate that 2-4 mM Ca2+ is suitable for proliferation and survival of osteoblasts, whereas slightly higher concentrations (6-8 mM) favor osteoblast differentiation and matrix mineralization in both 2- and 3-dimensional cultures. Higher concentrations (>10 mM) are cytotoxic. Purely from the perspective of calcium deposition, higher concentrations lead to increased accumulation of Ca2+. Culturing cells in phosphate-containing gel in media with Ca2+ also leads to time-dependent formation of HAp in the gel. Considering the viability of embedded cells, culturing scaffolds in media with Ca2+ concentrations around 5mM is useful for both HAp deposition and osteoblast behavior.
authors.affiliation: Department of Orthopaedic Surgery, Keio University Hospital, 35 Shinanomachi, Shinjukuku, Tokyo 160-8582, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Maeno
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Niki
authors.forename: Hideo
authors.initials: H
authors.lastname: Matsumoto
authors.forename: Hideo
authors.initials: H
authors.lastname: Morioka
authors.forename: Taku
authors.initials: T
authors.lastname: Yatabe
authors.forename: Atsushi
authors.initials: A
authors.lastname: Funayama
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Toyama
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Taguchi
authors.forename: Junzo
authors.initials: J
authors.lastname: Tanaka
mesh_terms: D000818:Animals; D018786:Bone Substitutes; D002118:Calcium / Q000737:chemistry* / Q000494:pharmacology*; D002356:Cartilage / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry* / Q000494:pharmacology*; D024043:Collagen Type II / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D017886:Durapatite / Q000737:chemistry; D007477:Ions; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D018786:Bone Substitutes; D020099:Coated Materials, Biocompatible; D024043:Collagen Type II; D007477:Ions; D017886:Durapatite; D002118:Calcium