Generation of optimized preparations of bone morphogenetic proteins for bone regeneration
Correction of inherited skeletal abnormalities, traumas affecting wide bone areas and non-healing fractures require efficient bone formation and regeneration. Bone morphogenetic proteins (BMPs) are signaling molecules that play a crucial role in bone and cartilage formation and regeneration. Osteoi...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry
2016-12-01
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| Series: | The Ukrainian Biochemical Journal |
| Subjects: | |
| Online Access: | http://ukrbiochemjournal.org/wp-content/uploads/2016/12/Malysheva_6_16.pdf |
| _version_ | 1797422315926978560 |
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| author | Kh. V. Malysheva I. M. Spasyuk O. K. Pavlenko |
| author_facet | Kh. V. Malysheva I. M. Spasyuk O. K. Pavlenko |
| author_sort | Kh. V. Malysheva |
| collection | DOAJ |
| description | Correction of inherited skeletal abnormalities, traumas affecting wide bone areas and non-healing fractures require efficient bone formation and regeneration. Bone morphogenetic proteins (BMPs) are signaling molecules that play a crucial role in bone and cartilage formation and regeneration. Osteoinductive properties of existing hydroxyapatite-based osteoplastic materials are frequently insufficient for efficient bone regeneration, thus raising a requirement for novel matrices involving BMPs for highly efficient local induction of bone formation at the area of the bone defect. The aim of this study was conducting in vitro optimization of osteoinductive properties of recombinant BMPs preparations to be used in bone regenerative practice. Recombinant BMPs were produced in human embryonic kidney 293 cells upon their transfection or co-transfection with plasmids expressing BMP2 and BMP7 at different ratios. A quality of BMP preps was validated based on their ability to induce in vitro osteoblast differentiation of C2C12 cells. Alkaline phosphatase that is widely used as a marker of osteoblast differentiation was measured spectrophotometrically. We found that the most effective inducer of osteoblast differentiation was recombinant BMP preparation produced upon cotransfection of 85% of BMP2 and 15% of BMP7 plasmids, that is most likely due to generation of conditions most favorable for formation of BMP2/7 heterodimers. Frozen BMP2/7 preparations stored for 3 h in experimental setup and for several weeks in routine work do not lose their osteoinductive properties compared with freshly prepared BMP2/7 preparations and can be successfully used for generation of highly efficient bone regenerative matrices. |
| first_indexed | 2024-03-09T07:30:26Z |
| format | Article |
| id | doaj.art-957be7b7873b42fdb8133b370c21cd81 |
| institution | Directory Open Access Journal |
| issn | 2409-4943 2413-5003 |
| language | English |
| last_indexed | 2024-03-09T07:30:26Z |
| publishDate | 2016-12-01 |
| publisher | National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry |
| record_format | Article |
| series | The Ukrainian Biochemical Journal |
| spelling | doaj.art-957be7b7873b42fdb8133b370c21cd812023-12-03T06:27:14ZengNational Academy of Sciences of Ukraine, Palladin Institute of BiochemistryThe Ukrainian Biochemical Journal2409-49432413-50032016-12-01886879710.15407/ubj88.06.087Generation of optimized preparations of bone morphogenetic proteins for bone regenerationKh. V. Malysheva0 I. M. Spasyuk1O. K. Pavlenko2Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv; Insitute of Animal Biology, National Academy of Agrarian Sciences of Ukraine, Lviv;Insitute of Animal Biology, National Academy of Agrarian Sciences of Ukraine, Lviv; Insitute of Animal Biology, National Academy of Agrarian Sciences of Ukraine, Lviv;Ivan Franko National University of Lviv, UkraineIvan Franko National University of Lviv, UkraineCorrection of inherited skeletal abnormalities, traumas affecting wide bone areas and non-healing fractures require efficient bone formation and regeneration. Bone morphogenetic proteins (BMPs) are signaling molecules that play a crucial role in bone and cartilage formation and regeneration. Osteoinductive properties of existing hydroxyapatite-based osteoplastic materials are frequently insufficient for efficient bone regeneration, thus raising a requirement for novel matrices involving BMPs for highly efficient local induction of bone formation at the area of the bone defect. The aim of this study was conducting in vitro optimization of osteoinductive properties of recombinant BMPs preparations to be used in bone regenerative practice. Recombinant BMPs were produced in human embryonic kidney 293 cells upon their transfection or co-transfection with plasmids expressing BMP2 and BMP7 at different ratios. A quality of BMP preps was validated based on their ability to induce in vitro osteoblast differentiation of C2C12 cells. Alkaline phosphatase that is widely used as a marker of osteoblast differentiation was measured spectrophotometrically. We found that the most effective inducer of osteoblast differentiation was recombinant BMP preparation produced upon cotransfection of 85% of BMP2 and 15% of BMP7 plasmids, that is most likely due to generation of conditions most favorable for formation of BMP2/7 heterodimers. Frozen BMP2/7 preparations stored for 3 h in experimental setup and for several weeks in routine work do not lose their osteoinductive properties compared with freshly prepared BMP2/7 preparations and can be successfully used for generation of highly efficient bone regenerative matrices.http://ukrbiochemjournal.org/wp-content/uploads/2016/12/Malysheva_6_16.pdfalkaline phosphatasebone morphogenetic proteinsbone regenerationmouse mesenchymal stem cellsosteogenesis |
| spellingShingle | Kh. V. Malysheva I. M. Spasyuk O. K. Pavlenko Generation of optimized preparations of bone morphogenetic proteins for bone regeneration The Ukrainian Biochemical Journal alkaline phosphatase bone morphogenetic proteins bone regeneration mouse mesenchymal stem cells osteogenesis |
| title | Generation of optimized preparations of bone morphogenetic proteins for bone regeneration |
| title_full | Generation of optimized preparations of bone morphogenetic proteins for bone regeneration |
| title_fullStr | Generation of optimized preparations of bone morphogenetic proteins for bone regeneration |
| title_full_unstemmed | Generation of optimized preparations of bone morphogenetic proteins for bone regeneration |
| title_short | Generation of optimized preparations of bone morphogenetic proteins for bone regeneration |
| title_sort | generation of optimized preparations of bone morphogenetic proteins for bone regeneration |
| topic | alkaline phosphatase bone morphogenetic proteins bone regeneration mouse mesenchymal stem cells osteogenesis |
| url | http://ukrbiochemjournal.org/wp-content/uploads/2016/12/Malysheva_6_16.pdf |
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