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An ectopic study of apatite-coated silk fibroin scaffolds seeded with AdBMP-2 modified canine bMSCs

Identifieur interne : 004308 ( Main/Exploration ); précédent : 004307; suivant : 004309

An ectopic study of apatite-coated silk fibroin scaffolds seeded with AdBMP-2 modified canine bMSCs

Auteurs : Kaige Lü [République populaire de Chine] ; Ling Xu [République populaire de Chine] ; Lunguo Xia [République populaire de Chine] ; Yilin Zhang [République populaire de Chine] ; Xiuli Zhang [République populaire de Chine] ; David L. Kaplan [États-Unis] ; Xinquan Jiang [République populaire de Chine] ; Fuqiang Zhang [République populaire de Chine]

Source :

RBID : PMC:3406182

Abstract

The present study was undertaken to evaluate ectopic new bone formation effects of apatite-coated silk fibroin scaffolds (mSS) seeded with adenovirus mediated bone morphogenic protein-2 gene (AdBMP-2) transduced canine bone marrow stromal cells (bMSCs) in nude mice. In this study, bMSCs derived from canine were cultured and transduced with AdBMP-2, adenovirus mediated enhanced green fluorescent protein gene (AdEGFP) in vitro. Osteogenic differentiation of bMSCs was determined by alkaline phosphatase (ALP) activity analysis, and the transcript levels for BMP-2, osteopontin (OPN), osteocalcin (OCN) and bone sialoprotein (BSP) genes via real-time quantitative PCR (RT-qPCR) analysis. The ectopic bone formation effects of mSS seeded with AdBMP-2 modified bMSCs were evaluated through histological and histomorphological analysis 4, 8 and 12 weeks post-operation in nude mice. ALP activity was statistically increased in AdBMP-2 group, when compared with control groups. The mRNA expression of BMP-2, OPN, OCN and BSP also statistically up-regulated 6, 9 days after AdBMP-2 transduction. Significantly higher bone volume was achieved in AdBMP-2-transduced bMSCs/mSS constructs than that of AdEGFP-transduced bMSCs/mSS or bMSCs/mSS groups at 4, 8 and 12 weeks (P<0.01). These results demonstrated that mSS seeded with AdBMP-2-transduced canine bMSCs can promote ectopic new bone formation and maturation in nude mice suggesting the potential of this silk scaffold based tissue engineered bone for further bone regeneration studies in canine models.


Url:
DOI: 10.1163/092050610X552861
PubMed: 21294971
PubMed Central: 3406182


Affiliations:


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<p id="P1">The present study was undertaken to evaluate ectopic new bone formation effects of apatite-coated silk fibroin scaffolds (mSS) seeded with adenovirus mediated bone morphogenic protein-2 gene (AdBMP-2) transduced canine bone marrow stromal cells (bMSCs) in nude mice. In this study, bMSCs derived from canine were cultured and transduced with AdBMP-2, adenovirus mediated enhanced green fluorescent protein gene (AdEGFP) in vitro. Osteogenic differentiation of bMSCs was determined by alkaline phosphatase (ALP) activity analysis, and the transcript levels for BMP-2, osteopontin (OPN), osteocalcin (OCN) and bone sialoprotein (BSP) genes via real-time quantitative PCR (RT-qPCR) analysis. The ectopic bone formation effects of mSS seeded with AdBMP-2 modified bMSCs were evaluated through histological and histomorphological analysis 4, 8 and 12 weeks post-operation in nude mice. ALP activity was statistically increased in AdBMP-2 group, when compared with control groups. The mRNA expression of BMP-2, OPN, OCN and BSP also statistically up-regulated 6, 9 days after AdBMP-2 transduction. Significantly higher bone volume was achieved in AdBMP-2-transduced bMSCs/mSS constructs than that of AdEGFP-transduced bMSCs/mSS or bMSCs/mSS groups at 4, 8 and 12 weeks (
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