Cell sheet transplantation of cultured mesenchymal stem cells enhances bone formation in a rat nonunion model

Nakamura, A.; Akahane, Manabu; Shigematsu, Hideki; Tadokoro, Mika; Morita, Yusuke; Ohgushi, Hajime; Dohi, Yoshiko; Imamura, Tomoaki; Tanaka, Yasuhito

doi:10.1016/j.bone.2009.08.048

Cited by 157 publications

(166 citation statements)

References 36 publications

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“…Although it might not be sufficiently robust, the use of osteogenic CSs represents a valuable approach not only for bone replacement but also for bone regeneration. 16,17,20 In this sense, we intended to move forward to the creation of a CS-based 3D construct and speculated that adding endothelial cells within the stacked osteogenic CSs would increase the amount of new tissue formed by promoting in vivo vascularization of the construct.…”

Section: Discussionmentioning

confidence: 99%

Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Pirraco

Iwata

Yoshida

et al. 2014

Laboratory Investigation

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Addressing the problem of vascularization is of vital importance when engineering three-dimensional (3D) tissues. Endothelial cells are increasingly used in tissue-engineered constructs to obtain prevascularization and to enhance in vivo neovascularization. Rat bone marrow stromal cells were cultured in thermoresponsive dishes under osteogenic conditions with human umbilical vein endothelial cells (HUVECs) to obtain homotypic or heterotypic cell sheets (CSs). Cells were retrieved as sheets from the dishes after incubation at 20 1C. Monoculture osteogenic CSs were stacked on top of homotypic or heterotypic CSs, and subcutaneously implanted in the dorsal flap of nude mice for 7 days. The implants showed mineralized tissue formation under both conditions. Transplanted osteogenic cells were found at the new tissue site, demonstrating CS bone-inductive effect. Perfused vessels, positive for human CD31, confirmed the contribution of HUVECs for the neovascularization of coculture CS constructs. Furthermore, calcium quantification and expression of osteocalcin and osterix genes were higher for the CS constructs, with HUVECs demonstrating the more robust osteogenic potential of these constructs. This work demonstrates the potential of using endothelial cells, combined with osteogenic CSs, to increase the in vivo vascularization of CS-based 3D constructs for bone tissue engineering purposes.

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Section: Discussionmentioning

confidence: 99%

Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Pirraco

Iwata

Yoshida

et al. 2014

Laboratory Investigation

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“…*, P < 0.05. able approaches for the repair or regeneration of damaged bone [22,23]. Tissue engineering and regenerative medicine have shown great promise in promoting bone repair using MSC sheets alone or in combination with implants wrapped in MSC sheets [13,24]. Compared with the traditional method of directly seeding cells in scaffolds, the cell sheet technique makes it possible to deliver a much larger number of cells for efficient tissue regeneration [9].…”

Section: Discussionmentioning

confidence: 99%

“…Recent studies have shown that the cell sheet technique is effective for the treatment of myocardial infarction [10], tendon defects [9] and periodontitis [11]. The cell sheet technique contributes to bone formation in cases of osseointegration [12], nonunion [13], various bone defects [14,15] and spinal interbody fusion [16].…”

mentioning

confidence: 99%

Restoration of rat calvarial defects by poly(lactide-co-glycolide)/hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes

Wang

Guo

et al. 2011

Chin. Sci. Bull.

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A composite construct comprising of a bone mesenchymal stem cell (BMSC) sheet, plasmid DNA, encoding human bone morphogenic protein-2 (hBMP-2), and poly(lactide-co-glycolide)/hydroxyapatite (PLGA/HA) sponge was designed and employed in the restoration of rat calvarial defects. To improve gene transfection efficiency, a cationic chitosan derivative, N,N,N,-trimethyl chitosan chloride (TMC), was employed as the vector. The TMC/DNA complexes had a transfection efficiency of 13% in rat BMSCs, resulting in heterogeneous hBMP-2 expression in a 10-d culture period in vitro. In vivo culture of the composite constructs was performed by implantation into rat full-thickness calvarial defects, using constructs lacking pDNA-hBMP-2 or BMSC sheets as controls. Significantly higher heterogeneous expression of hBMP-2 was detected in vivo at 2 weeks for the cell sheet/DNA complex/scaffold constructs, compared with the constructs lacking pDNA-hBMP-2 or BMSC sheets. New bone formation was evident as early as 4 weeks in the experimental constructs. At 8 weeks, partial bridging of calvarial defects was observed in the experimental constructs, which was significantly better than the constructs lacking pDNA-hBMP-2 or BMSC sheets. Therefore, the combination of the PLGA/HA scaffold with BMSC sheets and gene therapy vectors is effective at enhancing bone formation.gene therapy, bone regeneration, bone marrow stem cells, poly(lactide-co-glycolide), BMP-2 Citation:Li D, Wang W, Guo R, et al. Restoration of rat calvarial defects by poly(lactide-co-glycolide)/hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes.

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“…Institutional guidelines for the care and use of laboratory animals have been observed. We previously reported the method of bone marrow cell preparation [11,18]. Briefly, bone marrow cells were obtained from the femur shafts of 7-week-old male Fischer 344 rats.…”

Section: Bone Marrow Cell Preparation and Cell Culturementioning

confidence: 99%

“…Bone marrow-derived MSCs (BMSCs) have the potential for self-renewal and differentiation into several cell types after culture expansion in vitro and in vivo, including osteoblastic, chondrocytic, adipocytic and neuronal cells [3][4][5][6][7]. Differentiation into osteoblastic lineage cells after subculture in osteogenic-induced media, containing dexamethasone (Dex), ascorbic acid phosphate and β-glycerophosphate (β-GP), has been shown for rat and human BMSCs [8][9][10][11]. The osteogenic phenotypic markers, alkaline phosphatase (ALP) and osteocalcin (OC), can be detected by days 10 -12 in passage 1 (P1)-cultured cells, including mineralization [12].…”

Section: Introductionmentioning

confidence: 99%

Increased osteogenesis with hydroxyapatite constructs combined with serially-passaged bone marrow-derived mesenchymal stem cells

Akahane

Ueha²,

Shimizu³

et al. 2012

SCD

Self Cite

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We have previously reported on both the osteogenic potential of hydroxyapatite (HA) combined with bone marrow-derived mesenchymal stem cells (BMSCs) and a method involving osteogenic matrix cell sheet transplantation of BMSCs. In the present study, we assessed the osteogenic potential of serially-passaged BMSCs, both in vitro and in vivo. We also assessed whether an additional cell-loading technique can regain the osteogenic potential of the constructs combined with serially-passaged BMSCs. The present study revealed that passage (P) 1 cells cultured in osteogenic-induced medium showed strong positive staining for alkaline phosphatase (ALP) and Alizarin Red S, whereas P3 cells showed faint staining for ALP, with no Alizarin Red S staining. Staining of P1, P2 and P3 cells were progressively weaker, indicating that the osteogenic potential of the serially-passaged rat BMSCs is lost after P3 in vitro. The in vivo study showed that little bone formation was observed in the HA constructs seeded with P3 cells, 4 weeks after subcutaneous implantation. However, P3 cell/HA constructs which had increased cell-loading showed abundant bone formation within the pores of the HA construct. ALP and osteocalcin mRNA expression in these constructs was significantly higher than that of constructs with regular cell-seeding. The present study indicates that the osteogenic potential of the constructs with serially-passaged BMSCs is increased by additional cell-loading. This method can be applied to cases requiring hard tissue reconstruction, where BMSCs require serial expansion of cells.

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Cell sheet transplantation of cultured mesenchymal stem cells enhances bone formation in a rat nonunion model

Cited by 157 publications

References 36 publications

Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Restoration of rat calvarial defects by poly(lactide-co-glycolide)/hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes

Increased osteogenesis with hydroxyapatite constructs combined with serially-passaged bone marrow-derived mesenchymal stem cells

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