3D-cultured small size adipose-derived stem cell spheroids promote bone regeneration in the critical-sized bone defect rat model

Yamada, Yutaro; Okano, Tadashi; Orita, Kumi; Makino, Tomomi; Shima, Fumiaki; Nakamura, Hiroaki

doi:10.1016/j.bbrc.2022.03.027

Cited by 15 publications

(13 citation statements)

References 31 publications

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“…The mineralization increase in 3D models after exposure to NPs may be related to the greater cell surface capable of contacting NPs when compared to the monolayer (2D), enhancing the stimulatory effects of TiO 2 NPs [ 39 ]. This is consistent with previous studies that reveal that 3D osteoblasts models when exposed to TiO 2 NPs, compared to monolayer cells, induce the secretion of vascular endothelial growth factor (VEGF), activating a cascade of events resulting in higher type I collagen production [ 39 , 40 , 41 , 42 , 43 ]. Bone mineralization is the first step for implant osseointegration and begins when collagen I acts as a three-dimensional scaffold for hydroxyapatite deposition [ 44 ].…”

Section: Discussionsupporting

confidence: 92%

The Effects of Titanium Dioxide Nanoparticles on Osteoblasts Mineralization: A Comparison between 2D and 3D Cell Culture Models

et al. 2023

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Although several studies assess the biological effects of micro and titanium dioxide nanoparticles (TiO2 NPs), the literature shows controversial results regarding their effect on bone cell behavior. Studies on the effects of nanoparticles on mammalian cells on two-dimensional (2D) cell cultures display several disadvantages, such as changes in cell morphology, function, and metabolism and fewer cell–cell contacts. This highlights the need to explore the effects of TiO2 NPs in more complex 3D environments, to better mimic the bone microenvironment. This study aims to compare the differentiation and mineralized matrix production of human osteoblasts SAOS-2 in a monolayer or 3D models after exposure to different concentrations of TiO2 NPs. Nanoparticles were characterized, and their internalization and effects on the SAOS-2 monolayer and 3D spheroid cells were evaluated with morphological analysis. The mineralization of human osteoblasts upon exposure to TiO2 NPs was evaluated by alizarin red staining, demonstrating a dose-dependent increase in mineralized matrix in human primary osteoblasts and SAOS-2 both in the monolayer and 3D models. Furthermore, our results reveal that, after high exposure to TiO2 NPs, the dose-dependent increase in the bone mineralized matrix in the 3D cells model is higher than in the 2D culture, showing a promising model to test the effect on bone osteointegration.

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Section: Discussionsupporting

confidence: 92%

The Effects of Titanium Dioxide Nanoparticles on Osteoblasts Mineralization: A Comparison between 2D and 3D Cell Culture Models

et al. 2023

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“…In a recent study, it was shown that the right kind of stem cells is essential for neuromuscular regeneration and that neural crest stem cells have a specific advantage and an ability to treat peripheral nerve injuries by promoting re-innervation [ 10 ]. Due to their capacity for regeneration, adipose-derived stem cells are helpful in treating bone and cartilage abnormalities [ 12 ]. The fundamental mechanism was examined previously.…”

Section: Discussionmentioning

confidence: 99%

“…Despite mesenchymal stem cells’ ability to function well, the therapeutic potential of these cells may be greatly hindered by the gradual loss of stem cell properties that happens with repeated passages [ 11 ]. Small adipose-derived stem cell spheroids that were cultivated in three dimensions without a scaffold persisted in in vitro and in vivo settings and encouraged bone healing [ 12 ]. As a stem cell–based biomaterial therapy, neurosphere-medium spheroids should offer excellent clinical potential for bone and tissue regenerative therapies [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

Effects of Enamel Matrix Derivative on Cell Spheroids Made of Stem Cells Obtained from the Gingiva on Osteogenic Differentiation

Hwa

Lee

et al. 2023

Medicina

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Background and Objectives: A derivative of the enamel matrix was used to speed up periodontal regeneration, including the formation of new cementum, alveolar bone, and periodontal ligament. In this study, human gingiva-derived stem cell–derived cell spheroids were used to assess the effects of an enamel matrix derivative on cell viability, osteogenic differentiation, and mineralization. Materials and Methods: Human gingiva-derived stem cells were used to create spheroids, which were then coupled with unloaded control groups and an enamel matrix derivative at a final concentration of 2.7, 27, 270, and 2700 μg/mL. The morphological examination of the created stem cell spheroids took place on days 1, 3, 5, and 7. The Live/Dead Kit assay was used to determine the qualitative viability of cells on days 3 and 7. Using the Cell Counting Kit-8, the quantitative vitality of the cell spheroids was assessed on days 1, 3, and 5. On days 7 and 14, alkaline phosphatase activity assays and Alizarin Red S staining were carried out to examine the osteogenic differentiation of the cell spheroids. RUNX2 and COL1A1 expression levels on days 7 and 14 were determined using real-time polymerase chain reaction. Results: The added enamel matrix derivative at the tested concentrations did not significantly alter the morphology of the applied stem cells’ well-formed spheroids on day 1. On days 3 and 7, the majority of the spheroids’ cells fluoresced green while they were being cultivated. Alkaline phosphatase activity data revealed a substantial rise in the 2700 μg/mL group on day 7 when compared to the unloaded control (p < 0.05). On days 7 and 14, calcium deposits were distinctly seen in each group. In the 27 and 2700 μg/mL groups, the treatment with the enamel matrix derivative resulted in noticeably higher values for the Alizarin Red S staining (p < 0.05). qPCR results showed that adding an enamel matrix derivative to the culture of the 27 μg/mL group raised the level of RUNX2 mRNA expression. Conclusions: These results lead us to the conclusion that a derivative of the enamel matrix may be used to promote osteogenic differentiation in stem cell spheroids.

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“…Three-dimensional (3D) cell aggregates can imitate the natural microenvironment [ 8 ]. Small adipose-derived stem cell spheroids cultured in scaffold-free three-dimensional condition survived and promoted bone regeneration under in vitro and in vivo conditions [ 9 ]. Small injectable adipose stem cell spheroids are suggested to be a less invasive treatment alternative for the treatment of bony defects [ 9 ].…”

Section: Introductionmentioning

confidence: 99%

“…Small adipose-derived stem cell spheroids cultured in scaffold-free three-dimensional condition survived and promoted bone regeneration under in vitro and in vivo conditions [ 9 ]. Small injectable adipose stem cell spheroids are suggested to be a less invasive treatment alternative for the treatment of bony defects [ 9 ]. The three-dimensional spheroid culture of stem cells increased the production of key inflammatory regulators [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Assessment of the Impacts of Centipeda minima (L.) on Cell Viability, and Osteogenic Differentiation of Mesenchymal Stem Cell Spheroids

Lee

Uddin³

et al. 2022

Medicina

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Background and Objectives: Centipeda minima (L.) is a well-known and traditional pharmaceutical that has been utilized to treat different conditions controlling rhinitis, soothe pain, and decrease swelling. We assessed the impacts of Centipeda minima (L.) extricates (CMTs) on the osteogenic differentiation of cell spheroids made of human-bone-marrow-derived mesenchymal stem cells. Materials and Methods: Mesenchymal stem cells (MSCs) in spheroid 3D culture were generated and propagated in the presence of CMTs ranging from 0 to 1 μg/mL. Cell morphology was measured on Days 1, 3, 5, and 7. The quantitative cellular viability was evaluated on Days 1, 3, 5, and 7. Alkaline phosphatase activity assays were designed to measure the osteogenic differentiation of mesenchymal stem cell spheroids on Day 7. Alizarin Red S staining was performed to investigate the mineralization of cell spheroids on Days 7 and 14. Real-time polymerase chain reactions were used to measure the expression levels of RUNX2 and COL1A1 on Day 14. Western blot techniques were performed to identify the protein expression of Runt-related transcription factor 2 and type I collagen. Results: The control group’s mesenchymal stem cells displayed a spheroid shape. There was no noticeable change in morphology with the addition of CMTs at final concentrations of 0.001, 0.01, 0.1, and 1 μg/mL compared with the untreated (control) group. The application of CMTs did not induce a significant change in cell viability. The relative alkaline phosphatase activity values in the 0.001, 0.01, 0.1, and 1 μg/mL CMT groups were 114.4% ± 8.2%, 130.6% ± 25.3%, 87.8% ± 3.4%, and 92.1% ± 6.8%, respectively, considering a control of 100% (100.0% ± 17.9%). On Day 14, calcium deposits were clearly observed in each group. The relative values of Alizarin Red S staining in the 0.001, 0.01, 0.1, and 1 μg/mL CMT groups were 100.1% ± 8.9%, 105.9% ± 0.0%, 109.7% ± 19.1%, and 87.0% ± 40.9%, respectively, considering a control of 100% (100.0% ± 28.7%). The addition of CMT significantly increased RUNX2 expression in the 0.01 μg/mL group and COL1A1 in the 0.001 and 0.01 μg/mL groups. Normalization of protein expression showed that the addition of CMTs significantly increased type I collagen expression in the 0.001, 0.01, and 1 μg/mL groups. Conclusions: In conclusion, CMTs influence the osteogenic differentiation of bone-marrow-derived mesenchymal stem cells and the use of CMTs may positively influence the osteogenic differentiation of cell spheroids.

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3D-cultured small size adipose-derived stem cell spheroids promote bone regeneration in the critical-sized bone defect rat model

Cited by 15 publications

References 31 publications

The Effects of Titanium Dioxide Nanoparticles on Osteoblasts Mineralization: A Comparison between 2D and 3D Cell Culture Models

The Effects of Titanium Dioxide Nanoparticles on Osteoblasts Mineralization: A Comparison between 2D and 3D Cell Culture Models

Effects of Enamel Matrix Derivative on Cell Spheroids Made of Stem Cells Obtained from the Gingiva on Osteogenic Differentiation

Assessment of the Impacts of Centipeda minima (L.) on Cell Viability, and Osteogenic Differentiation of Mesenchymal Stem Cell Spheroids

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