Masako Fujioka-Kobayashi1,2, Kosaku Sawada1,3, Eizaburo Kobayashi1, Benoit Schaller1, Yufeng Zhang4, Richard J Miron5,6. 1. Department of Operative, Preventive and Pediatric Dentistry, University of Bern, Bern, Switzerland. 2. Department of Oral Surgery, Institute of Biomedical Sciences, Tokushima University, Tokushima, Japan. 3. Advanced Research Center, School of Life Dentistry at Niigata, The Nippon Dental University, Niigata, Japan. 4. Department of Oral Implantology, Wuhan University, Wuhan, China. 5. Department of Periodontology, School of Dental Medicine, University of Bern. 6. Department of Periodontology, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, FL.
Abstract
BACKGROUND: Bone morphogenetic protein 9 (BMP9) has previously been characterized as one of the most osteogenic growth factors of the BMP family. To the best of the authors' knowledge, previous experiments have only used adenovirus transfection (gene therapy). With the recent development of recombinant human BMP9 (rhBMP9), the present study investigates the osteopromotive potential of BMP9 versus rhBMP2 when loaded onto collagen membranes. METHODS: ST2 stromal bone marrow cells were seeded onto: 1) control; 2) low-dose rhBMP2 (10 ng/mL); 3) high-dose rhBMP2 (100 ng/mL); 4) low-dose rhBMP9 (10 ng/mL); and 5) high-dose rhBMP9 (100 ng/mL) porcine collagen membranes. The following parameters were compared among groups: 1) cell adhesion (at 8 hours); 2) cell proliferation (at 1, 3, and 5 days); 3) real-time polymerase chain reaction for genes encoding runt-related transcription factor 2; 4) alkaline phosphatase (ALP); 5) bone sialoprotein ([BSP] at 3 and 14 days); and 6) alizarin red staining (at 14 days). RESULTS: rhBMP2 and rhBMP9 demonstrated little effect on cell attachment and proliferation; however, pronounced increases were observed in osteoblast differentiation. All groups significantly induced ALP messenger RNA (mRNA) levels at 3 days and BSP levels at 14 days; however, high-dose rhBMP9 showed significantly higher values compared with all other groups for ALP levels (five-fold increase at 3 days and two-fold increase at 14 days). Alizarin red staining further revealed both concentrations of rhBMP9 induced up to three-fold more staining compared with rhBMP2. CONCLUSIONS: Results indicate that the combination of collagen membranes with rhBMP9 induced significantly higher ALP mRNA expression and alizarin red staining compared with rhBMP2. These findings suggest that rhBMP9 may be a suitable growth factor for future regenerative procedures in bone biology.
BACKGROUND:Bone morphogenetic protein 9 (BMP9) has previously been characterized as one of the most osteogenic growth factors of the BMP family. To the best of the authors' knowledge, previous experiments have only used adenovirus transfection (gene therapy). With the recent development of recombinant humanBMP9 (rhBMP9), the present study investigates the osteopromotive potential of BMP9 versus rhBMP2 when loaded onto collagen membranes. METHODS: ST2 stromal bone marrow cells were seeded onto: 1) control; 2) low-dose rhBMP2 (10 ng/mL); 3) high-dose rhBMP2 (100 ng/mL); 4) low-dose rhBMP9 (10 ng/mL); and 5) high-dose rhBMP9 (100 ng/mL) porcine collagen membranes. The following parameters were compared among groups: 1) cell adhesion (at 8 hours); 2) cell proliferation (at 1, 3, and 5 days); 3) real-time polymerase chain reaction for genes encoding runt-related transcription factor 2; 4) alkaline phosphatase (ALP); 5) bone sialoprotein ([BSP] at 3 and 14 days); and 6) alizarin red staining (at 14 days). RESULTS: rhBMP2 and rhBMP9 demonstrated little effect on cell attachment and proliferation; however, pronounced increases were observed in osteoblast differentiation. All groups significantly induced ALP messenger RNA (mRNA) levels at 3 days and BSP levels at 14 days; however, high-dose rhBMP9 showed significantly higher values compared with all other groups for ALP levels (five-fold increase at 3 days and two-fold increase at 14 days). Alizarin red staining further revealed both concentrations of rhBMP9 induced up to three-fold more staining compared with rhBMP2. CONCLUSIONS: Results indicate that the combination of collagen membranes with rhBMP9 induced significantly higher ALP mRNA expression and alizarin red staining compared with rhBMP2. These findings suggest that rhBMP9 may be a suitable growth factor for future regenerative procedures in bone biology.
Entities:
Keywords:
Bone morphogenetic protein 2; bone regeneration; osteogenesis; recombinant proteins
Authors: Behnoush Khorsand; Satheesh Elangovan; Liu Hong; Alexander Dewerth; Michael S D Kormann; Aliasger K Salem Journal: AAPS J Date: 2017-01-10 Impact factor: 3.603
Authors: Masako Fujioka-Kobayashi; Benoit Schaller; Nikola Saulacic; Benjamin E Pippenger; Yufeng Zhang; Richard J Miron Journal: Clin Exp Dent Res Date: 2017-02-09