Osteogenic Differentiation of Recombinant Adeno-Associated Virus 2-Transduced Murine Mesenchymal Stem Cells and Development of an Immunocompetent Mouse Model for Ex Vivo Osteoporosis Gene Therapy

Kumar, Sanjay; Mahendra, Gandham; Nagy, Tim R.; Ponnazhagan, Selvarangan

doi:10.1089/hum.2004.15.ft-5

Cited by 13 publications

(15 citation statements)

References 47 publications

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“…BMSCs belong to the osteogenic stem cells and have the potential to transform into various cell lines. They can achieve cross-system differentiation under certain external environment conditions and stimulating factors, and as they have the characteristics of being convenient to obtain and easy to culture, they are a good source of seed cells for tissue engineering (5,13,14). BMSCs consist of two types of osteogenic precursor cells: i) Oriented osteogenic precursor cells (OOPCs) that can perform oriented differentiation into osteogenic cells without induction; ii) induced osteogenic precursor cells (IOPCs) that can only perform oriented differentiation into osteogenic cells with induction by specific factors.…”

Section: Discussionmentioning

confidence: 99%

Effect of recombinant adeno-associated BMP-4/7 fusion gene on the biology of BMSCs

Yuan

Zhang

2012

Molecular Medicine Reports

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The aim of the study was to investigate the effect of recombinant adeno-associated virus (AAV) with morphogenetic protein 4/7 fusion gene on the ossification of rabbit bone marrow stromal cells (BMSCs). The genes BMP-4 and BMP-7 were obtained through one-step reverse transcriptase polymerase chain reaction from human placental cells. The BMP-4/7 fusion gene was generated through recombination. The rabbit BMSCs were transfected with the recombinant AAV vectors carrying AAV-BMP-4/7 with various multiplicity of infection (MOI) values. Expression of fusion gene BMP-4/7 protein was determined by the ELISA method. The ossification of cells was evaluated by observing alkaline phosphatase (ALP) and osteocalcin (OC) activity after transfection for 7 and 14 days. We successfully constructed the recombinant AAV with the BMP-4/7 fusion gene. A 29-20 kDa protein was shown by SDS-PAGE electrophoresis following transfection with AAV-BMP-4/7. The fusion protein BMP-4/7 in BMSCs transferred by AAV showed a positive correlation with various MOI values. There was significantly higher ALP and OC activity in the AAV-BMP-4/7 transfection groups than in the AAV-EGFP groups (t(ALP)=896.88, P<0.001, t(OC)=543.24, P<0.01). The fusion gene BMP-4/7 is capable of enhancing the expression of BMPs and possesses significant ossification activity through AAV.

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Section: Discussionmentioning

confidence: 99%

Effect of recombinant adeno-associated BMP-4/7 fusion gene on the biology of BMSCs

Yuan

Zhang

2012

Molecular Medicine Reports

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“…A few researchers previously showed in vivo bone formation using rAAV-expressing BMPs. [34][35][36] Gene therapy for bone regeneration has not been studied sufficiently with rAAV, probably because of the weak expression from these vectors. However, with several modalities available to enhance gene expression from rAAV, 21,25 further research is warranted.…”

Section: Discussionmentioning

confidence: 99%

“…In mesenchymal cells, for example, a reporter gene was expressed for up to 150 days in an explant culture of articular cartilage 26 and for up to 4 weeks in a monolayer cell culture. 36 In contrast, the expression of a reporter gene in vivo persisted for >1.5 years in muscles, 37 whereas b-galactosidase expression was achieved for just 10 days in rat knee articular cartilage. 26 Mesenchymal tissues tend to express recombinant proteins for a shorter time than other organs, although the number of studies is limited; gene expression is effective for 2 days to 2 weeks after injection into bone or cartilage.…”

Section: Discussionmentioning

confidence: 99%

Biological activation of bone‐related biomaterials by recombinant adeno‐associated virus vector

Nasu

Ito

Tsutsumi

et al. 2009

Journal Orthopaedic Research

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Gene therapy is a promising clinical tool that is no longer limited as a method to supplement genetic deficits, but rather is considered reliable for delivering proteins to specific tissues or cells. Recombinant adeno-associated virus (rAAV) vector is one of the most potent gene transfer vehicles. Many biomaterials have been used in reconstructive surgery, but their biological inactivity has limited their use. To overcome shortcomings of available bone-related biomaterials, we investigated the combination of rAAV with biomaterials. Taking advantage of the method of lyophilizing rAAV onto biomaterials, we showed that an rAAV coating successfully induced b-galactosidase protein expression by rat fibroblasts on hydroxyapatite, b-tricalcium phosphate, and titanium alloy in vitro. b-Galactosidase expression was detected for 8 weeks after implantation of rAAV-coated hydroxyapatite into rat back muscles in vivo. A coating of bone morphogenetic protein-2-expressing rAAV induced significant de novo bone formation on hydroxyapatite in rat back muscles. Our study demonstrates that the combination of lyophilized rAAV and biomaterials presents a promising strategy for bone regenerative medicine. ß

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“…Administration of BMP‐2 was able to increase MSC number, osteogenic activity and decline in rate of osteoblast apoptosis in a number of studies. Kumar et al 2004 engineered murine MSC to over‐express BMP‐2 and such modified MSCs showed significant osteoblastic differentiation compared to naïve MSCs in culture. MSCs, over‐expressing BMP‐2 has been shown to restore bone defects in aged rats [Tsuda et al, 2005].…”

Section: Therapeutic Application Of Msc In Bone Diseasesmentioning

confidence: 99%

“…However, it is very difficult to predict if bone marrow stromal cells harbor differentially committed MSC progenitors or their fate is determined by diverse isolation and culture conditions. Since its inception MSCs have become an important tool in tissue engineering and regenerative medicine ranging from generation of cardiomyocytes after a myocardial infarction to restore skeletal damage pertaining to osteoporosis, fracture, and osteolytic bone metastases [Kumar et al, 2004; Rabin et al, 2007; Fritz et al, 2008; Chanda et al, 2009]. Besides, HSC and endothelial progenitor cells (EPCs) enhance MSC function by promoting osteoblastic differentiation and neovascularization, respectively, at the injury site(s).…”

mentioning

confidence: 99%

Therapeutic potential of adult bone marrow‐derived mesenchymal stem cells in diseases of the skeleton

Chanda

Kumar

Ponnazhagan

2010

J of Cellular Biochemistry

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115

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Mesenchymal stem cells (MSCs) are the most popular among the adult stem cells in tissue engineering and regenerative medicine. Since their discovery and functional characterization in the late sixties and early seventies, MSCs or MSC-like cells have been obtained from various mesodermal and non-mesodermal tissues, although majority of the therapeutic applications involved bone marrow derived MSCs. Based on its mesenchymal origin, it was predicted earlier that MSCs only can differentiate into mesengenic lineages like bone, cartilage, fat or muscle. However, varied isolation and cell culturing methods identified subsets of MSCs in the bone marrow which not only differentiated into mesenchymal lineages, but also into ectodermal and endodermal derivatives.

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Osteogenic Differentiation of Recombinant Adeno-Associated Virus 2-Transduced Murine Mesenchymal Stem Cells and Development of an Immunocompetent Mouse Model for Ex Vivo Osteoporosis Gene Therapy

Cited by 13 publications

References 47 publications

Effect of recombinant adeno-associated BMP-4/7 fusion gene on the biology of BMSCs

Effect of recombinant adeno-associated BMP-4/7 fusion gene on the biology of BMSCs

Biological activation of bone‐related biomaterials by recombinant adeno‐associated virus vector

Therapeutic potential of adult bone marrow‐derived mesenchymal stem cells in diseases of the skeleton

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