The effect of calcium ion concentration on osteoblast viability, proliferation and differentiation in monolayer and 3D culture.
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.
Maeno
Shinichi
S
Department of Orthopaedic Surgery, Keio University Hospital, 35 Shinanomachi, Shinjukuku, Tokyo 160-8582, Japan.
Niki
Yasuo
Y
Matsumoto
Hideo
H
Morioka
Hideo
H
Yatabe
Taku
T
Funayama
Atsushi
A
Toyama
Yoshiaki
Y
Taguchi
Tetsushi
T
Tanaka
Junzo
J
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*
D018786:Bone Substitutes; D020099:Coated Materials, Biocompatible; D024043:Collagen Type II; D007477:Ions; D017886:Durapatite; D002118:Calcium