Regeneration of calvarial defects with escherichia coli-derived rhBMP-2 adsorbed in PLGA Membrane

Mitsuaki Ono, Wataru Sonoyama, Kazuki Nema, Emilio satoshi Hara, Yasutaka Oida, Hai Thanh Pham, Katushi Yamamoto, Kazuo Hirota, Kazushige Sugama, Walter Sebald, Takuo Kuboki

Research output: Contribution to journalArticle

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Abstract

Objective:Escherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria. Materials and Methods: First, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 μg/μl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology. Results: Release-profile testing showed that PLGA membrane could retain 94% of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 μg/μl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-μg/μl dose of E-BMP-2. Conclusion: PLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.

Original languageEnglish
Pages (from-to)367-376
Number of pages10
JournalCells Tissues Organs
Volume198
Issue number5
DOIs
Publication statusPublished - 2014

Fingerprint

Regeneration
Escherichia coli
Membranes
Osteogenesis
Bone Regeneration
X-Ray Microtomography
polylactic acid-polyglycolic acid copolymer
Osteocalcin
Mesenchymal Stromal Cells
Skull
Histology
Enzyme-Linked Immunosorbent Assay
Staining and Labeling
Calcium
Messenger RNA
Proteins

Keywords

  • Bone regeneration
  • Ectopic bone formation
  • Escherichia coli-derived recombinant human bone morphogenetic protein-2
  • Polylactide-co-glycolide

ASJC Scopus subject areas

  • Anatomy
  • Histology
  • Medicine(all)

Cite this

Regeneration of calvarial defects with escherichia coli-derived rhBMP-2 adsorbed in PLGA Membrane. / Ono, Mitsuaki; Sonoyama, Wataru; Nema, Kazuki; Hara, Emilio satoshi; Oida, Yasutaka; Pham, Hai Thanh; Yamamoto, Katushi; Hirota, Kazuo; Sugama, Kazushige; Sebald, Walter; Kuboki, Takuo.

In: Cells Tissues Organs, Vol. 198, No. 5, 2014, p. 367-376.

Research output: Contribution to journalArticle

Ono, M, Sonoyama, W, Nema, K, Hara, E, Oida, Y, Pham, HT, Yamamoto, K, Hirota, K, Sugama, K, Sebald, W & Kuboki, T 2014, 'Regeneration of calvarial defects with escherichia coli-derived rhBMP-2 adsorbed in PLGA Membrane', Cells Tissues Organs, vol. 198, no. 5, pp. 367-376. https://doi.org/10.1159/000356947
Ono, Mitsuaki ; Sonoyama, Wataru ; Nema, Kazuki ; Hara, Emilio satoshi ; Oida, Yasutaka ; Pham, Hai Thanh ; Yamamoto, Katushi ; Hirota, Kazuo ; Sugama, Kazushige ; Sebald, Walter ; Kuboki, Takuo. / Regeneration of calvarial defects with escherichia coli-derived rhBMP-2 adsorbed in PLGA Membrane. In: Cells Tissues Organs. 2014 ; Vol. 198, No. 5. pp. 367-376.
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abstract = "Objective:Escherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria. Materials and Methods: First, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 μg/μl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology. Results: Release-profile testing showed that PLGA membrane could retain 94{\%} of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 μg/μl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-μg/μl dose of E-BMP-2. Conclusion: PLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.",
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T1 - Regeneration of calvarial defects with escherichia coli-derived rhBMP-2 adsorbed in PLGA Membrane

AU - Ono, Mitsuaki

AU - Sonoyama, Wataru

AU - Nema, Kazuki

AU - Hara, Emilio satoshi

AU - Oida, Yasutaka

AU - Pham, Hai Thanh

AU - Yamamoto, Katushi

AU - Hirota, Kazuo

AU - Sugama, Kazushige

AU - Sebald, Walter

AU - Kuboki, Takuo

PY - 2014

Y1 - 2014

N2 - Objective:Escherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria. Materials and Methods: First, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 μg/μl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology. Results: Release-profile testing showed that PLGA membrane could retain 94% of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 μg/μl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-μg/μl dose of E-BMP-2. Conclusion: PLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.

AB - Objective:Escherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria. Materials and Methods: First, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 μg/μl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology. Results: Release-profile testing showed that PLGA membrane could retain 94% of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 μg/μl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-μg/μl dose of E-BMP-2. Conclusion: PLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.

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KW - Ectopic bone formation

KW - Escherichia coli-derived recombinant human bone morphogenetic protein-2

KW - Polylactide-co-glycolide

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