In vitro bone-like nodules generated from patient-derived iPSCs recapitulate pathological bone phenotypes

Kawai, Shunsuke; Yokota, Hiroyuki; Sunaga, Junko; Alev, Cantas; Nagata, Shinji; Nishio, Makoto; Hada, M.; Koyama, Yuko; Uemura, Maya; Sekiguchi, Kenichi; Maekawa, Hirotsugu; Ikeya, Motoji; Tamaki, Sakura; Jin, Young-Woo; Harada, Yuka; Fukiage, Kenichi; Adachi, Taiji; Matsuda, Shinya; Toguchida, Junya

doi:10.1038/s41551-019-0410-7

Cited by 61 publications

(53 citation statements)

References 60 publications

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“…When the EBs were coated, confluence was expected at 70-80% ( Figure 1a-d); in CTR8#P12, since it was the most copious clone, retinoic acid was added as described previously ( Figure 2) [16]. From the phase microscope images, we observed that, in the first days of differentiation, the speed of expansion and differentiation was faster (Figure 3), which then appeared to slow (Figure 4).…”

Section: Resultssupporting

confidence: 52%

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New Application of Osteogenic Differentiation from HiPS Stem Cells for Evaluating the Osteogenic Potential of Nanomaterials in Dentistry

Tetè

Capparè

Gherlone

2020

IJERPH

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Objective: HiPS stem cells are commonly used for the study of medical disorders. The laboratory in which this study was conducted uses these cells for examining the treatment and cure of neurodegenerative diseases. Bone regeneration poses the greatest challenge for an oral surgeon both in terms of increased implant osseointegration and reducing bone healing times. The aim of this study was to validate the protocol in the literature to produce and then test in vitro osteoblasts with different nanomaterials to simulate bone regeneration. Method: hiPS clones (#2, #4, and #8) were differentiated into an osteoblast cell culture tested for alizarin red staining and for alkaline phosphatase testing at 14, 21 and 28 days, after the cells were plated. Results: The cells showed diffuse positivity under alizarin red staining and the alkaline phosphatase (ALP)-test, showing small formations of calcium clusters. Conclusion: Despite the limitations of our study, it is a starting point for further protocols, laying a solid foundation for research in the field of bone regeneration through the use of stem cells.

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Section: Resultssupporting

confidence: 52%

“…Another variance in our protocol is the use of retinoic acid in control 8 (the clone with the largest cell representation [16]). In our study, clone 8 + RA initially showed a more accelerated trend than clone 8 − RA, but a less orderly organization and a slower trend.…”

Section: Discussionmentioning

confidence: 99%

New Application of Osteogenic Differentiation from HiPS Stem Cells for Evaluating the Osteogenic Potential of Nanomaterials in Dentistry

Tetè

Capparè

Gherlone

2020

IJERPH

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“…Bone and periodontal regeneration is a complicated process that requires the cooperation of myriad cells [25]. Mesenchymal stem cells (MSCs) or osteoblastic precursors recruit and differentiate into osteoblasts when periapical tissues of tooth surrounded by alveolar bone and periodontal ligament regenerate after pulpal infection and inflammation at periapical lesions due to tooth decay [1,2]. Still, the biomineralization process of MSCs is largely governed by local environment [3][4][5][6][7][8].…”

Section: Discussionmentioning

confidence: 99%

“…When periapical tissues, which are alveolar bone and periodontal ligament that surrounds the root of the tooth, regenerate after pulp (soft tissue inside of tooth) infection and periapical lesion due to tooth decay, mesenchymal stem cells (MSCs) or osteoblastic precursors recruit and differentiate into osteoblasts during healing process [1,2]. e biomineralization process of MSCs is largely governed by local environment such as stiffness, stress relaxation, nanotopology, and chemicals [3][4][5][6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Initial Cytotoxicity of Mineral Trioxide Aggregate (MTA) during Setting on Human Mesenchymal Stem Cells

Sooyoun

Lee

2019

Advances in Materials Science and Engineering

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Bone-marrow-derived human mesenchymal stem cells (hMSCs) which are important cell source for hard tissue regeneration stay near periapical lesions of tooth, playing an essential role in periodontal regeneration. Since the biomineralization process of MSCs is largely governed by the initial local environment, it is crucial to know the biological effects of dental bioceramic (mineral trioxide aggregate (MTA)) right after implantation. The purpose of this study was to evaluate the initial cytotoxicity of 4 different commercially available MTA materials (Endocem MTA, Ortho MTA, ProRoot MTA, and MTA Angelus) against hMSCs during or after setting using extracts of materials. The materials were mixed separately and placed into disk-shaped Teflon split molds (10 mm diameter and 2 mm thickness), and the sample discs were separated and eluted in the culture medium for 24 h. The extracts were exposed to hMSCs, and cytotoxicity was evaluated by the WST assay. In the present study, all 4 MTA products tested showed severe cytotoxicity at 100% and 50% extract, while 25% and 12.5% revealed 30∼100% depending on the MTA products. Endocem MTA showed severe cytotoxicity at 12.5% extract, while others showed relatively higher cell viability compared to Endocem MTA. Images of live and dead cells represented less live cells at 25% and 12.5%, confirming cell viability assay. Therefore, careful consideration of the concentration of MTA extracts is necessary, especially when applying MTA to the elderly patients to maintain the viability of hMSCs.

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“…Mesenchymal tissues, including bone and cartilage, are generated by mesoderm (PM and LM) and neuroectoderm-derived neural crest [20] ( Figure S1). Although it is unclear how RA precisely regulates the lineage commitment of iPSC, RA was previously applied to osteogenic induction protocols for mouse [15,16,21] and human iPSCs [22]. In addition, RA-treated EBs formed from mouse embryonic stem cells (ESCs) produce pre-somatic mesoderm and neural crest cells, both of which provide immature mesenchymal cells that can differentiate into osteoblasts and chondrocytes [23].…”

Section: Discussionmentioning

confidence: 99%

In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells

Limraksasin

Kondo

Zhang

et al. 2020

IJMS

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Cell condensation and mechanical stimuli play roles in osteogenesis and chondrogenesis; thus, they are promising for facilitating self-organizing bone/cartilage tissue formation in vitro from induced pluripotent stem cells (iPSCs). Here, single mouse iPSCs were first seeded in micro-space culture plates to form 3-dimensional spheres. At day 12, iPSC spheres were subjected to shaking culture and maintained in osteogenic induction medium for 31 days (Os induction). In another condition, the osteogenic induction medium was replaced by chondrogenic induction medium at day 22 and maintained for a further 21 days (Os-Chon induction). Os induction produced robust mineralization and some cartilage-like tissue, which promoted expression of osteogenic and chondrogenic marker genes. In contrast, Os-Chon induction resulted in partial mineralization and a large area of cartilage tissue, with greatly increased expression of chondrogenic marker genes along with osterix and collagen 1a1. Os-Chon induction enhanced mesodermal lineage commitment with brachyury expression followed by high expression of lateral plate and paraxial mesoderm marker genes. These results suggest that combined use of micro-space culture and mechanical stimuli facilitates hybrid bone/cartilage tissue formation from iPSCs, and that the bone/cartilage tissue ratio in iPSC constructs could be manipulated through the induction protocol.

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In vitro bone-like nodules generated from patient-derived iPSCs recapitulate pathological bone phenotypes

Cited by 61 publications

References 60 publications

New Application of Osteogenic Differentiation from HiPS Stem Cells for Evaluating the Osteogenic Potential of Nanomaterials in Dentistry

New Application of Osteogenic Differentiation from HiPS Stem Cells for Evaluating the Osteogenic Potential of Nanomaterials in Dentistry

Initial Cytotoxicity of Mineral Trioxide Aggregate (MTA) during Setting on Human Mesenchymal Stem Cells

In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells

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