Histological evaluations of apatite-fiber scaffold cultured with mesenchymal stem cells by implantation at rat subcutaneous tissue

Suzuki, Kitaru; Nagata, Kouji; Yokota, Tomohiro; Honda, Michiyo; Aizawa, Mamoru

doi:10.3233/bme-171656

Cited by 4 publications

(4 citation statements)

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“…Regarding the efficacy of h BMMSCs, subcutaneous ectopic bone formation models are commonly used by CBT [40,41]. Previous reports have demonstrated that murine or human bone marrow stromal cells seeded on calcium phosphate (CaP) stimulate the bone formation implanted subcutaneously in immune-compromised mice [42].…”

Section: Discussionmentioning

confidence: 99%

Preclinical Studies of the Biosafety and Efficacy of Human Bone Marrow Mesenchymal Stem Cells Pre-Seeded into β-TCP Scaffolds after Transplantation

et al. 2018

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Background: Cell-Based Therapies (CBT) constitute a valid procedure for increasing the quantity and quality of bone in areas with an inadequate bone volume. However, safety and efficacy should be investigated prior to clinical application. The objective of this study was to evaluate the biodistribution, safety and osteogenic capacity of bone marrow-derived human mesenchymal stem cells (hBMMSCs) pre-seeded into β-tricalcium phosphate (TCP) and implanted into NOD/SCID mice at subcutaneous and intramuscular sites. Methods: hBMMSCs were isolated, characterized and then cultured in vitro on a porous β-TCP scaffold. Cell viability and attachment were analyzed and then hBMMSCs seeded constructs were surgically placed at subcutaneous and intramuscular dorsal sites into NOD/SCID mice. Acute and subchronic toxicity, cell biodistribution and efficacy were investigated. Results: There were no deaths or adverse events in treated mice during the 48-hour observation period, and no toxic response was observed in mice. In the 12-week subchronic toxicity study, no mortalities, abnormal behavioral symptoms or clinical signs were observed in the saline control mice or the hBMMSCs/β-TCP groups. Finally, our results showed the bone-forming capacity of hBMMSCs/β-TCP since immunohistochemical expression of human osteocalcin was detected from week 7. Conclusions: These results show that transplantation of hBMMSCs/β-TCP in NOD/SCID mice are safe and effective, and might be applied to human bone diseases in future clinical trials.

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Section: Discussionmentioning

confidence: 99%

Preclinical Studies of the Biosafety and Efficacy of Human Bone Marrow Mesenchymal Stem Cells Pre-Seeded into β-TCP Scaffolds after Transplantation

et al. 2018

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“…Thus, there is a significant technical need of the development for preparing satisfactory prosthetic bone in an extracorporeal setting using a three-dimensional culture method [ 9 , 16 , 17 ]. In the present study, we attempted to prepare prosthetic bone extracorporeally by culturing mouse MC3T3-E1 cells (as a substitute for osteoblasts) in a radial-flow bioreactor with porous β-TCP or HA beads, which are clinically used for transplantation [ 18 – 20 ].…”

Section: Discussionmentioning

confidence: 99%

Bio-artificial bone formation model with a radial-flow bioreactor for implant therapy—comparison between two cell culture carriers: porous hydroxyapatite and β-tricalcium phosphate beads

et al. 2018

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Bone grafting is necessary before dental implant treatment in patients with jaw bone defects. Currently, autologous bone grafting is a major burden on the patient. However, it is impossible to form a sufficient foundation for the implant with a bone-filling agent alone. It is, therefore, necessary to prepare hybrid artificial bone tissue containing osteoblasts and osteoclasts. In this study, mouse MC3T3-E1 pre-osteoblast cells and human embryonic-derived osteoblastic cell line hFOB1.19 were cultured in radial-flow bioreactors (RFB) to form three-dimensional artificial bone filled with porous beads of β-tricalcium phosphate (β-TCP) or hydroxyapatite (HA)—which are clinically used as bone-filling agents—as cell culture carriers. When circulation culturing was performed in the growth medium for the first 10–12 days, glucose consumption was increased in the cultures with HA beads in comparison to the cultures with β-TCP beads. When cultured in the differentiation culture medium during the second half of the culture period, the glucose consumption decreased in the culture with HA beads. A DNA microarray analysis suggested that osteogenesis progressed fast in three-dimensional culture filled with HA beads and that partly differentiation into osteoblasts was prominent in cultures with β-TCP beads. In the growth process of MC3T3-E1 cells, the vitamin A metabolism was also activated, the synthesis and degradation of retinoic acid was enhanced, and the metabolism of the same process decreased at the end of differentiation in three-dimensional cultures. Three-dimensional circulation culture in RFB is considered to be useful for the formation of hybrid bio-artificial bone tissue.Electronic supplementary materialThe online version of this article (10.1007/s13577-018-0218-x) contains supplementary material, which is available to authorized users.

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“…In our previous study, bone formation was confirmed in vivo when AFS and pre-osteodifferentiated cells were implanted. 24) Ohgushi et al 25) conducted an experiment to evaluate the bone-forming ability of HAp ceramics cultured with RBMCs by implanting them at rat subcutaneous tissue. As a result, the formation of new bone was confirmed in the ceramics.…”

Section: Biological Evaluations Of Implanted Samplesmentioning

confidence: 99%

Living reactions of tissue-engineered bone derived from apatite-fiber scaffold in rat subcutaneous tissues

Suzuki

Honda

Matsuura

et al. 2022

J. Ceram. Soc. Japan

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We synthesized apatite fibers (AFs) and fabricated a porous hydroxyapatite scaffold (apatite-fiber scaffold; AFS). A tissue-engineered bone involving a three-dimensional structure was constructed by placing AFS in a radial-flow bioreactor (RFB) to culture rat bone marrow mesenchymal stem cells (RBMCs). In this study, we examined whether the tissue-engineered bone derived from the AFS has osteogenic differentiation potential leading to bone-forming ability in vivo in the subcutaneous tissue where bone formation does not occur naturally due to the absence of osteoblasts. The reconstructed tissue-engineered bone was implanted subcutaneously in rat tissue for 4 weeks. The AFS alone was implanted as a control. After implantation, some histological evaluations, i.e., hematoxylin and eosin (HE), alkaline phosphatase (ALP) staining, and immunofluorescence staining for osteocalcin (OC), were performed for the harvested samples. In addition, quantitative evaluation was also performed to determine the ALP activity normalized for the DNA content of the harvested samples. The HE staining revealed that both AFSs (control) and implanted tissue-engineered bone (RFB-bone) were biocompatible, and did not induce inflammation or immunological rejection in vivo. The ALP activity in the RFB-bone was significantly higher than that in the control. Immunofluorescent staining showed the expression of OC in RFB-bone. Therefore, we conclude that the tissue-engineered bone derived from AFS may have the boneforming ability in vivo in the absence of osteoblasts. These results may provide valuable insights into the design of tissue-engineered bone for clinical applications.

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Histological evaluations of apatite-fiber scaffold cultured with mesenchymal stem cells by implantation at rat subcutaneous tissue

Abstract: These findings suggest that AFSs are a conducive microenvironment for bone regeneration and are well tolerated in vivo. The results provide valuable baseline data for the design of implant studies using tissue-engineered bone constructed by RFB.

Cited by 4 publications

References 14 publications

Preclinical Studies of the Biosafety and Efficacy of Human Bone Marrow Mesenchymal Stem Cells Pre-Seeded into β-TCP Scaffolds after Transplantation

Preclinical Studies of the Biosafety and Efficacy of Human Bone Marrow Mesenchymal Stem Cells Pre-Seeded into β-TCP Scaffolds after Transplantation

Bio-artificial bone formation model with a radial-flow bioreactor for implant therapy—comparison between two cell culture carriers: porous hydroxyapatite and β-tricalcium phosphate beads

Living reactions of tissue-engineered bone derived from apatite-fiber scaffold in rat subcutaneous tissues

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