Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid Culture

Octacalcium phosphate (OCP) has been shown to exhibit an osteogenic property and, therefore, has been utilized recently as a bone substitute, clinically. However, the stimulatory capacity for induced pluripotent stem (iPS) cells is not known. This study investigated whether OCP enhances osteoblastic...

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Main Authors: Yuki Sugai, Ryo Hamai, Yukari Shiwaku, Takahisa Anada, Kaori Tsuchiya, Tai Kimura, Manami Tadano, Kensuke Yamauchi, Tetsu Takahashi, Hiroshi Egusa, Osamu Suzuki
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Biomimetics
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Online Access:https://www.mdpi.com/2313-7673/10/4/205
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author Yuki Sugai
Ryo Hamai
Yukari Shiwaku
Takahisa Anada
Kaori Tsuchiya
Tai Kimura
Manami Tadano
Kensuke Yamauchi
Tetsu Takahashi
Hiroshi Egusa
Osamu Suzuki
author_facet Yuki Sugai
Ryo Hamai
Yukari Shiwaku
Takahisa Anada
Kaori Tsuchiya
Tai Kimura
Manami Tadano
Kensuke Yamauchi
Tetsu Takahashi
Hiroshi Egusa
Osamu Suzuki
author_sort Yuki Sugai
collection DOAJ
description Octacalcium phosphate (OCP) has been shown to exhibit an osteogenic property and, therefore, has been utilized recently as a bone substitute, clinically. However, the stimulatory capacity for induced pluripotent stem (iPS) cells is not known. This study investigated whether OCP enhances osteoblastic differentiation of three-dimensionally cultured spheroids of iPS cells compared to hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP). Mouse iPS cells were mixed with smaller (less than 53 μm) or larger (300–500 μm) sizes of calcium phosphate (CaP) granules and cultured in a laboratory-developed oxygen-permeable culture chip under minimizing hypoxia for up to 21 days. Osteoblastic differentiation was estimated by the cellular alkaline phosphatase (ALP) activities. The degree of supersaturation (DS) with respect to CaP phases was determined from the media chemical compositions. Incubated CaP materials were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The culture promoted well the formation of hybrid spheroids of CaP materials and iPS cells regardless of the type of materials and their granule sizes. The ALP activity of OCP was about 1.5 times higher than that of β-TCP and HA in smaller granule sizes. FTIR, XRD, and DS analyses showed that larger OCP granules tended to hydrolyze to HA slightly faster than smaller granules with time while HA and β-TCP materials tended to remain unchanged. In conclusion, the results suggest that OCP enhances the osteogenic differentiation of iPS cells more than HA and β-TCP through a mechanism of hydrolyzing to HA. This inherent material property of OCP is essential for enhancing the osteoblastic differentiation of iPS cells.
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spelling doaj-art-ae358f42c36e45cbbb9704ec01f55b332025-08-20T02:24:43ZengMDPI AGBiomimetics2313-76732025-03-0110420510.3390/biomimetics10040205Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid CultureYuki Sugai0Ryo Hamai1Yukari Shiwaku2Takahisa Anada3Kaori Tsuchiya4Tai Kimura5Manami Tadano6Kensuke Yamauchi7Tetsu Takahashi8Hiroshi Egusa9Osamu Suzuki10Division of Craniofacial Function Engineering (Division of Biomaterials Science and Engineering), Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Craniofacial Function Engineering (Division of Biomaterials Science and Engineering), Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Craniofacial Function Engineering (Division of Biomaterials Science and Engineering), Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Craniofacial Function Engineering (Division of Biomaterials Science and Engineering), Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Craniofacial Function Engineering (Division of Biomaterials Science and Engineering), Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Craniofacial Function Engineering (Division of Biomaterials Science and Engineering), Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Pediatric Dentistry, Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Oral and Maxillofacial Reconstructive Surgery, Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Oral and Maxillofacial Reconstructive Surgery, Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Molecular and Regenerative Prosthodontics, Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanDivision of Craniofacial Function Engineering (Division of Biomaterials Science and Engineering), Tohoku University Graduate School of Dentistry, Sendai 980-8575, JapanOctacalcium phosphate (OCP) has been shown to exhibit an osteogenic property and, therefore, has been utilized recently as a bone substitute, clinically. However, the stimulatory capacity for induced pluripotent stem (iPS) cells is not known. This study investigated whether OCP enhances osteoblastic differentiation of three-dimensionally cultured spheroids of iPS cells compared to hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP). Mouse iPS cells were mixed with smaller (less than 53 μm) or larger (300–500 μm) sizes of calcium phosphate (CaP) granules and cultured in a laboratory-developed oxygen-permeable culture chip under minimizing hypoxia for up to 21 days. Osteoblastic differentiation was estimated by the cellular alkaline phosphatase (ALP) activities. The degree of supersaturation (DS) with respect to CaP phases was determined from the media chemical compositions. Incubated CaP materials were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The culture promoted well the formation of hybrid spheroids of CaP materials and iPS cells regardless of the type of materials and their granule sizes. The ALP activity of OCP was about 1.5 times higher than that of β-TCP and HA in smaller granule sizes. FTIR, XRD, and DS analyses showed that larger OCP granules tended to hydrolyze to HA slightly faster than smaller granules with time while HA and β-TCP materials tended to remain unchanged. In conclusion, the results suggest that OCP enhances the osteogenic differentiation of iPS cells more than HA and β-TCP through a mechanism of hydrolyzing to HA. This inherent material property of OCP is essential for enhancing the osteoblastic differentiation of iPS cells.https://www.mdpi.com/2313-7673/10/4/205hybrid spheroidsiPS celloctacalcium phosphateinorganic ionsosteogenic differentiation
spellingShingle Yuki Sugai
Ryo Hamai
Yukari Shiwaku
Takahisa Anada
Kaori Tsuchiya
Tai Kimura
Manami Tadano
Kensuke Yamauchi
Tetsu Takahashi
Hiroshi Egusa
Osamu Suzuki
Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid Culture
Biomimetics
hybrid spheroids
iPS cell
octacalcium phosphate
inorganic ions
osteogenic differentiation
title Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid Culture
title_full Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid Culture
title_fullStr Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid Culture
title_full_unstemmed Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid Culture
title_short Effect of Octacalcium Phosphate on Osteogenic Differentiation of Induced Pluripotent Stem Cells in a 3D Hybrid Spheroid Culture
title_sort effect of octacalcium phosphate on osteogenic differentiation of induced pluripotent stem cells in a 3d hybrid spheroid culture
topic hybrid spheroids
iPS cell
octacalcium phosphate
inorganic ions
osteogenic differentiation
url https://www.mdpi.com/2313-7673/10/4/205
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