Utility of tricalcium phosphate and osteogenic matrix cell sheet constructs for bone defect reconstruction

Ueha, Tomoyuki; Akahane, Manabu; Shimizu, Takamasa; Uchihara, Yoshinobu; Morita, Yusuke; Nitta, Naoya; Kido, Akira; Inagaki, Yusuke; Kawate, Kenji; Tanaka, Yasuhito

doi:10.4252/wjsc.v7.i5.873

Cited by 20 publications

(23 citation statements)

References 35 publications

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“…Various biomaterials have been developed to enhance the osteogenic differentiation of BMSCs in vitro . However, in vivo bone regeneration, especially in ectopic sites, with these systems has been inefficient and unsatisfactory, because of the random dispersal of the cells in the biological scaffold . Cell sheet technology, developed recently, could be a good solution for bone tissue engineering and, therefore, continues to attract significant research interest.…”

Section: Discussionmentioning

confidence: 99%

“…Cell sheet technology provides an effective strategy for using BMSCs, not only to minimize the destruction of the ECM, but also to improve cell retention at the transplant site and attachment to the scaffold . BMSC sheets wrapped around a scaffold showed more pronounced capacity for bone regeneration than did cells dispersed in the scaffold …”

Section: Introductionmentioning

confidence: 99%

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Enhancing bone regeneration by combining mesenchymal stem cell sheets with β‐TCP/COL‐I scaffolds

Lin

Shao

et al. 2017

J Biomed Mater Res

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The combination of bone marrow-derived mesenchymal stem cells (BMSCs) and biological scaffolds has been demonstrated to be a promising strategy for bone regeneration. However, this method does not result in satisfactory bone regeneration, because the BMSCs are dispersed in the biological scaffolds. The current study developed a new bone regeneration system, which combines synthetic porous three-dimensional scaffolds of β-TCP/COL-I composite with cultured osteogenic sheets of BMSCs. Activity of alkaline phosphatase (ALP), a marker of bone regeneration, was assayed in vitro using enzyme-linked immunosorbent assays and quantitative real-time polymerase chain reaction. In vivo bone regeneration was assayed in male nude mice. The study samples were BMSC sheet, scaffold/scattered BMSCs, scaffold/BMSC sheet, and scaffold alone. The samples were implanted dorsally in the mice. In vitro analysis showed that β-TCP/COL-I scaffold combined with BMSC sheets significantly upregulated both gene expression and protein levels of ALP, osteocalcin, and osteopontin. Histological and micro-computed tomography showed that the only implants that demonstrated new bone formation after 4 weeks were scaffold/BMSC sheet implants. These results underscore the crucial requirement of a synergistic effect of β-TCP/COL-I scaffolds and BMSC sheets. This could be a promising novel strategy for bone tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2037-2045, 2018.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Enhancing bone regeneration by combining mesenchymal stem cell sheets with β‐TCP/COL‐I scaffolds

Lin

Shao

et al. 2017

J Biomed Mater Res

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“…Unlike in methods that use trypsin/EDTA digestion, because the ECM in an osteogenic matrix cell sheet remains intact, they are suitable for skeletal reconstruction applications such as bone defects and fracture nonunions. Transplantation of an osteogenic matrix cell sheet includes both necessary features of cell transplantation and ECM transplantation, which together, can fill in gaps between bones and artificial bone, 11,12 as well as to unite artificial bones. PGs are a major component of the animal extracellular matrix and are formed of glycosaminoglycans (GAGs) covalently attached to the core proteins.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, BMSC populations containing MSCs are a good source for creating tissue-engineered bone. 11,12 …”

Section: Introductionmentioning

confidence: 99%

Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure

Akahane

Shimizu

Kira

et al. 2016

Bone & Joint Research

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ObjectivesTo assess the structure and extracellular matrix molecule expression of osteogenic cell sheets created via culture in medium with both dexamethasone (Dex) and ascorbic acid phosphate (AscP) compared either Dex or AscP alone.MethodsOsteogenic cell sheets were prepared by culturing rat bone marrow stromal cells in a minimal essential medium (MEM), MEM with AscP, MEM with Dex, and MEM with Dex and AscP (Dex/AscP). The cell number and messenger (m)RNA expression were assessed in vitro, and the appearance of the cell sheets was observed after mechanical retrieval using a scraper. β-tricalcium phosphate (β-TCP) was then wrapped with the cell sheets from the four different groups and subcutaneously implanted into rats.ResultsAfter mechanical retrieval, the osteogenic cell sheets from the MEM, MEM with AscP, and MEM with Dex groups appeared to be fragmented or incomplete structures. The cell sheets cultured with Dex/AscP remained intact after mechanical retrieval, without any identifiable tears. Culture with Dex/AscP increased the mRNA and protein expression of extracellular matrix proteins and cell number compared with those of the other three groups. More bridging bone formation was observed after transplantation of the β-TCP scaffold wrapped with cell sheets cultured with Dex/AscP, than in the other groups.ConclusionsThese results suggest that culture with Dex/AscP improves the mechanical integrity of the osteogenic cell sheets, allowing retrieval of the confluent cells in a single cell sheet structure. This method may be beneficial when applied in cases of difficult tissue reconstruction, such as nonunion, bone defects, and osteonecrosis.Cite this article: M. Akahane, T. Shimizu, T. Kira, T. Onishi, Y. Uchihara, T. Imamura, Y. Tanaka. Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure. Bone Joint Res 2016;5:569–576. DOI: 10.1302/2046-3758.511.BJR-2016-0013.R1.

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“…The released BMSCs were seeded onto 100-mm culture dishes at a density of 1 9 10 4 cells/cm 2 to generate OMCS [13,14,25]. Briefly, the cells were cultured in standard medium with 10 nM Dex and 0.28 mM AscP (cell sheet medium) for 14 days.…”

Section: Preparation Of the Omcs And B-tcp Ceramics Constructmentioning

confidence: 99%

Implantation of Bone Marrow Stromal Cell Sheets Derived from Old Donors Supports Bone Tissue Formation

Akahane

Shimizu

Inagaki

et al. 2017

Tissue Eng Regen Med

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The purpose of this study was to evaluate the osteogenesis ability of osteogenic matrix cell sheets (OMCS) derived from old donor cells. Bone marrow stromal cells (BMSC) were obtained from young (7-week-old) and old (1-yearold) Fischer344 rats donors and cultured with modified Eagle's medium (MEM group) alone or containing dexamethasone (Dex; 10 nM) and ascorbic acid phosphate (AscP; 0.28 mM) (Dex/AscP group). We prepared four in vitro experimental groups: (1) young MEM, (2) young Dex/AscP, (3) old MEM and (4) old Dex/AscP. Cell proliferation and osteogenic marker mRNA expression levels were evaluated in vitro. To assess bone formation in vivo, the cells of each group were combined with beta tricalcium phosphate (TCP) disks followed by implantation in recipient rats. The in vitro study showed significant differences in the mRNA expression of osteocalcin, ALP, and BMP2 between MEM and Dex/AscP groups. Bone formation following implantation was observed upon histological analyses of all groups. TCP combined with OMCS (OMCS/TCP group) resulted in enhanced bone formation compared to that following combination with BMSC (BMSC/ TCP). The osteocalcin content of the OMCS/TCP group 4 weeks after implantation was significantly higher than that in the BMSC/TCP construct for both young and old donors. The present study clearly indicated that OMCS could be generated from BMSCs of old as well as young donors using a mechanical retrieval method. Thus, through its usage of OMCS, this method may represent a potentially effective therapeutic option for cell-based therapy in elderly patients.

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Utility of tricalcium phosphate and osteogenic matrix cell sheet constructs for bone defect reconstruction

Abstract: This technique is an improvement over current methods, such as TCP substitution, and is useful for hard tissue reconstruction and inducing earlier bone union in defects.

Cited by 20 publications

References 35 publications

Enhancing bone regeneration by combining mesenchymal stem cell sheets with β‐TCP/COL‐I scaffolds

Enhancing bone regeneration by combining mesenchymal stem cell sheets with β‐TCP/COL‐I scaffolds

Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure

Implantation of Bone Marrow Stromal Cell Sheets Derived from Old Donors Supports Bone Tissue Formation

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