Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery

Kang, Qi; Mh, Sun; Cheng, Hongwei; Peng, Ying; Ag, Montag; At, Deyrup; Jiang, Wei; Hh, Luu; Luo, Jianxun; Jp, Szatkowski; Vanichakarn, Pantila; Jy, Park; Y, Li; Rc, Haydon; He, Tong‐Chuan

doi:10.1038/sj.gt.3302298

Cited by 519 publications

(735 citation statements)

References 83 publications

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“…The key step is the generation of recombinants in AdEasier cells. Once these recombinants are obtained, generation of adenoviruses in HEK-293 cells is virtually guaranteed [23][24][25][26][27] . Depending on the transfection efficiency of HEK-293 cells, one can expect titers in the initial lysate to be 10 8 -10 10 pfu ml À1 .…”

Section: Anticipated Resultsmentioning

confidence: 99%

A protocol for rapid generation of recombinant adenoviruses using the AdEasy system

et al. 2007

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Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We have developed an approach that simplifies the generation and production of such viruses called the AdEasy system. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, employing homologous recombination in bacteria rather than in eukaryotic cells. After transfection of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of GFP encoded by a gene incorporated into the viral backbone. This system has expedited the process of generating and testing recombinant adenoviruses for a variety of purposes. In this protocol, we describe the practical aspects of using the AdEasy system for generating recombinant adenoviruses. The full protocol usually takes 4-5 weeks to complete.

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Section: Anticipated Resultsmentioning

confidence: 99%

A protocol for rapid generation of recombinant adenoviruses using the AdEasy system

et al. 2007

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“…BMP antagonist, noggin inhibited BMP2 osteogenesis, however SMAD phosphorylation by BMP9 was not inhibited by exogenous noggin (Bergeron et al, 2009; Nakamura, Shinohara, Momozaki, Yoshimoto, & Noguchi, 2013; Wang et al, 2013). BMP3, a known BMP2 inhibitor, did not inhibit BMP9‐mediated bone formation (Kang et al, 2004). Further research utilizing rhBMP9 would benefit our understanding on the differential mechanisms that regulate osteogenesis in both BMP2 versus BMP9.…”

Section: Discussionmentioning

confidence: 99%

“…Although these previous studies were confirmed only utilizing BMP9‐adenovirus‐transfected cells not approved by the FDA (Cheng et al, 2003; Kang et al, 2004; Lamplot et al, 2013; Leblanc et al, 2011), our research group recently characterized the regenerative potential of rhBMP9 in comparison to rhBMP2. In two studies, it was found that rhBMP9 demonstrated up to 10 times greater osteogenic differentiation when compared to rhBMP2 (Fujioka‐Kobayashi et al, 2016a, 2016b).…”

Section: Introductionmentioning

confidence: 85%

“…Although recombinant human (rh)BMP2 is the most widely utilized recombinant BMP with Food and Drug Administration (FDA)'s approval (Bessa, Casal, & Reis, 2008a, 2008b; Carreira et al, 2014), many investigators remain surprised to learn that other BMPs have previously been characterized as more osteogenic than the currently approved BMP2 or BMP7 (Kang et al, 2004). In two pioneering study conducted over a decade ago investigating all 14 BMPs, it was found that BMP6 and BMP9 stimulated the highest alkaline phosphatase (ALP) expression in vitro (Cheng et al, 2003) and had greater potential for orthotropic ossification in vivo (Kang et al, 2004). In those studies, the bone‐inducing properties of all BMPs were reported using adenovirus transfection experiments (gene therapy), an area of research still not approved by the FDA for clinical use (Balmayor & van Griensven, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2

Fujioka‐Kobayashi

Benoit

Saulačić

et al. 2017

Clinical & Exp Dental Res

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The use of growth factors for the regeneration of soft and hard tissues has been utilized extensively in dental medicine over the past decade. Recently our group found that recombinant human bone morphogenetic protein 9 (rhBMP9) was more osteopromotive than recombinant human bone morphogenetic protein 2 (rhBMP2) when combined with a deprotenized bovine bone mineral bone grafting material. The aim of the present in vitro study was to evaluate the regenerative potential of an absorbable collagen sponge(ACS) specifically designed for extraction socket healing loaded with rhBMP9 when compared to rhBMP2. The adsorption and release kinetics of rhBMP2 and rhBMP9 were first investigated by enzyme‐linked immunosorbent assay quantification. Then, the cellular effects of stromal cell line (ST2) preosteoblasts were investigated utilizing four groups including rhBMP2 and rhBMP9 at both low(10 ng/ml) and high(100 ng/ml) concentrations loaded onto ACS. Cellular attachment(8 hours) and proliferation(1, 3, and 5 days) as well as osteoblast differentiation were investigated by real‐time polymerase chain reaction (PCR) at 3 and 14 days, alkaline phosphatase (ALP) activity at 7 days, and alizarin red staining at 14 days. ACS fully adsorbed both rhBMP2 and rhBMP9 that were slowly released up to 10 days. Although neither rhBMP2 nor rhBMP9 had any effects on cell attachment or proliferation, pronounced effects were observed on osteoblast differentiation. ALP activity was increased seven‐fold with rhBMP2‐high, whereas a marked 10‐fold and 20‐fold increase was observed with rhBMP9‐low and high loaded to ACS, respectively. Furthermore, mRNA levels of collagen1, ALP, bone sialoprotein, and osteocalcin were all significantly higher for rhBMP9 when compared to control or rhBMP2 groups. Alizarin red staining further confirmed that rhBMP9‐low and high demonstrated marked increases in mineralization potential when compared to rhBMP2‐high. The results demonstrate the marked effect of rhBMP9 on osteoblast differentiation when combined with ACS in comparison to rhBMP2 at doses as much as 10 times lower. Further in vivo studies are necessary to investigate whether the regenerative potential is equally as potent.

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“…Moreover, since BMPs are known inducers of osteogenic differentiation, 24 and a local increase of synthesis of several BMPs in the peri‐prosthetic tissue has been reported,23 we also stimulated cells with OM supplemented with either BMP‐2 or BMP‐6. Several BMPs, including BMP‐2 and BMP‐6, have shown positive effects on bone formation, fracture healing, and implant osseointegration in several in vitro and in vivo animal models 30, 31, 32, 33. BMP‐2 is even used in clinical practice for accelerated healing of fractures and for spinal fusions 30, 34.…”

Section: Discussionmentioning

confidence: 99%

Peri-prosthetic tissue cells show osteogenic capacity to differentiate into the osteoblastic lineage

et al. 2017

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During the process of aseptic loosening of prostheses, particulate wear debris induces a continuous inflammatory‐like response resulting in the formation of a layer of fibrous peri‐prosthetic tissue at the bone‐prosthesis interface. The current treatment for loosening is revision surgery which is associated with a high‐morbidity rate, especially in old patients. Therefore, less invasive alternatives are necessary. One approach could be to re‐establish osseointegration of the prosthesis by inducing osteoblast differentiation in the peri‐prosthetic tissue. Therefore, the aim of this study was to investigate the capacity of peri‐prosthetic tissue cells to differentiate into the osteoblast lineage. Cells isolated from peri‐prosthetic tissue samples (n = 22)−obtained during revision surgeries−were cultured under normal and several osteogenic culture conditions. Osteogenic differentiation was assessed by measurement of Alkaline Phosphatse (ALP), mineralization of the matrix and expression of several osteogenic genes. Cells cultured in osteogenic medium showed a significant increase in ALP staining (p = 0.024), mineralization of the matrix (p < 0.001) and ALP gene expression (p = 0.014) compared to normal culture medium. Addition of bone morphogenetic proteins (BMPs), a specific GSK3β inhibitor (GIN) or a combination of BMP and GIN to osteogenic medium could not increase ALP staining, mineralization, and ALP gene expression. In one donor, addition of GIN was required to induce mineralization of the matrix. Overall, we observed a high‐inter‐donor variability in response to osteogenic stimuli. In conclusion, peri‐prosthetic tissue cells, cultured under osteogenic conditions, can produce alkaline phosphatase and mineralized matrix, and therefore show characteristics of differentiation into the osteoblastic lineage. © 2016 The Authors. Journal of Orthopaedic Research published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 35:1732–1742, 2017.

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Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery

Cited by 519 publications

References 83 publications

A protocol for rapid generation of recombinant adenoviruses using the AdEasy system

A protocol for rapid generation of recombinant adenoviruses using the AdEasy system

Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2

Peri-prosthetic tissue cells show osteogenic capacity to differentiate into the osteoblastic lineage

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