BMP-2/4 and BMP-6/7 Differentially Utilize Cell Surface Receptors to Induce Osteoblastic Differentiation of Human Bone Marrow-derived Mesenchymal Stem Cells

Abstract

Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-β superfamily of growth factors and are used clinically to induce new bone formation. The purpose of this study was to evaluate receptor utilization by BMP-2, BMP-4, BMP-6, and BMP-7 in primary human mesenchymal stem cells (hMSC), a physiologically relevant cell type that probably mediates the in vivo effects of BMPs. RNA interference-mediated gene knockdown revealed that osteoinductive in are the type and and the type and and and to a in by of the in this receptor utilization and was to was to by the type used was the type receptor by and was by of of and that in The of and to and was by and in primary Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-β superfamily of growth factors and are used clinically to induce new bone formation. The purpose of this study was to evaluate receptor utilization by BMP-2, BMP-4, BMP-6, and BMP-7 in primary human mesenchymal stem cells (hMSC), a physiologically relevant cell type that probably mediates the in vivo effects of BMPs. RNA interference-mediated gene knockdown revealed that osteoinductive in are the type and and the type and and and to a in by of the in this receptor utilization and was to was to by the type used was the type receptor by and was by of of and that in The of and to and was by and in primary Bone morphogenetic proteins (BMPs) used bone morphogenetic mesenchymal stem human mesenchymal stem cell growth RNA used bone morphogenetic mesenchymal stem human mesenchymal stem cell growth RNA are members of the transforming growth factor-β superfamily of growth are of growth and and in and and BMP-7 and are members of this of growth factors and are used clinically to induce new bone in and bone and BMP-7 to the and the The of to induce a of the cells to the of and the of a to the are by of of type and type type and and type and revealed the of and type receptor of to receptor to of the type by type type the and the to the and factors to induce the of and to the osteoinductive of BMPs. the Bone and the that type and type cells type receptor and the receptor BMP-7 to and to to and BMP-7 and The of BMP-7 in the of was in cells a in type receptor BMP-7 and BMP-4, BMP-7 stem cells are a cell type that the human bone mesenchymal stem cell that this cell type to and bone the of BMPs. The study was to evaluate receptor utilization by osteoinductive BMP-2, BMP-4, BMP-6, and the of primary was to type type in to receptor utilization by The in type and type receptor the this type receptor receptor by and The in to in clinically relevant cell and and and mesenchymal stem cell growth and in and used of the cells and in and BMP-2, BMP-6, and BMP-7 in cells and are was was to the the of and the the of in was in to the in was the of was to the of was the to the was of in a and was and by a and gene was a and gene to the used in this study are and of gene and receptor knockdown was the of to the by The of receptor in and was the the human of used in to and and to the relevant are in of to of to the and a and by of to The of of gene in was the and to of the used to the type and and the type and and to the of and a a the a and the of of was to and cells in to of gene of BMP-2, in in receptor and gene was by the of receptor and the of of of gene in the to the to the of receptor knockdown and the of and the of and The the by and of a of and was and the to gene was by and are in the and in and and and the of that the proteins the receptor by a the of the receptor and was the by the and used in the cells The cells a of of of the of receptor a of cells and in in to of and a was by to a of and was and The was the of the and proteins and the in to the by cells of the receptor and to of the was a by and the of to cells of the of the and of the receptor of the and of BMP-4, BMP-6, and BMP-7 in of BMP-2, BMP-4, BMP-6, and BMP-7 to the of primary was the and The of was by the osteoinductive of the and was to the by of the of receptor and was of by The gene in the of in and that induce a gene in the of the to induce in by in in The was that induce of primary that the and of this of receptor utilization in was to the osteoinductive the of in and the of in primary The of receptor was and to was the of the and the type and type to and to and to and to of and was the of the to and of receptor in primary and human of and was by a of the the of receptor in are the of of of and of type and type in human are the of of of the receptor in human the receptor was and to was the type receptor in the of and The in to in to to and to of type receptor and and of type was the type receptor in in to in and in to in The of in was the the was receptor a of of and a of in to human to of receptor the receptor of receptor the of and by of the to by to receptor to and in of type type knockdown was by the of knockdown and to in to and to and to in of was probably in the of by the of to the knockdown of the type that the a the of type type and type are by of and primary a of of of the of type type of the was by the of are to the of receptor in to by in the of receptor by the of the the of of cells the was in the the was the and to of of type type by BMP-2, BMP-4, BMP-6, BMP-7 was a primary The knockdown of the type and the of to induce the of and by that type type are to in primary in the of the in this the effects by and of the receptor knockdown the effects of receptor knockdown and that the of the of receptor and in by the and and primary a of of of the of type type cells BMP-2, BMP-4, BMP-6, BMP-7 and of was by and to are the of receptor knockdown to the the of that to the are receptor knockdown the and are by in type receptor utilization of by and BMP-7 by and by and The and was a by knockdown of in and of and BMP-4, and and The and was a by of knockdown was of the and and that and are the primary type the of in the of and a of and that and the to of type receptor to and of in of by BMPs. the of this was the was of and was to the and BMP-7 a by in a of by and a BMP-7 The BMP-7 in human bone by and by of knockdown was of the BMPs. The of and knockdown was and knockdown the and receptor knockdown of and a of of and in cells BMP-2, BMP-4, BMP-6, of and of was by and to are the of gene receptor knockdown to the the of the was in the the was the and BMP-7 of and and of and the knockdown of in of that the of by of the type receptor was by the that knockdown a in the knockdown of in a of and that the of by of the type receptor in this was by the that knockdown in a in by and that and BMP-7 type receptor in the of and of type receptor utilization by the and in primary and in by of and of and in by of of and of of and to to the of and and BMP-4, the knockdown of in of that the by in a of a was to by the knockdown of in of that the of by in of and BMP-4, a the of that a of and by of and The of the and that and by that a of and in a of and and to and to this the to and in a and to in to the and of and in cells proteins and was by proteins to the receptor in was by of and was of that and of the was The of the the to the that are that of the and the of a to and of and and cells in The cells of of the of receptor and and by a of to of by and the of to the of by of of of of of of of of of relevant receptor the of that the are in the this of of in the by that of that and of the was The of the in this study that was the to the of type and formation. of a of and of of to in the the in of of the the in the of and and in the of type of and of this study was to evaluate receptor utilization the of in primary receptor knockdown revealed that the osteoinductive of in primary are the type and and the type and and to in this to the of of of the in cell a of in to of and and the in this cell type in cells growth of in primary human bone cells the and of receptor utilization in in in this study the of the of receptor members of the to the type was to to by was the type to osteoinductive in The of to a in of by are in the type receptor knockdown in receptor by and to of and and a of to and and was in cells of type by used to of and of the of cells of the receptor that a of receptor in cells to the of the to that a type of receptor to the in of the and a receptor of a of the type receptor and of the type and of the type receptor a that of a of by the type a in the of in cells and the to osteoinductive in primary members of the and to receptor and utilization of type in the of in a of The in are to induce of cells to BMP-7 effects of BMP-7 to of BMP-7 the BMP-6, was to transforming growth and the and of BMP-7 in the and of in a the of receptor receptor in human to the of in the of in the of receptor the that in receptor human in and in the in this study that BMP-2, BMP-4, BMP-6, and BMP-7 induce of primary in the to a of type type the effects of in receptor utilization and in the the in clinically relevant Bone morphogenetic proteins (BMPs) used bone morphogenetic mesenchymal stem human mesenchymal stem cell growth RNA used bone morphogenetic mesenchymal stem human mesenchymal stem cell growth RNA are members of the transforming growth factor-β superfamily of growth are of growth and and in and and BMP-7 and are members of this of growth factors and are used clinically to induce new bone in and bone and BMP-7 to the and the The of to induce a of the cells to the of and the of a to the are by of of type and type type and and type and revealed the of and type receptor of to receptor to of the type by type type the and the to the and factors to induce the of and to the osteoinductive of BMPs. the Bone and the that type and type cells type receptor and the receptor BMP-7 to and to to and BMP-7 and The of BMP-7 in the of was in cells a in type receptor BMP-7 and BMP-4, BMP-7 stem cells are a cell type that the human bone mesenchymal stem cell that this cell type to and bone the of BMPs. The study was to evaluate receptor utilization by osteoinductive BMP-2, BMP-4, BMP-6, and the of primary was to type type in to receptor utilization by The in type and type receptor the this type receptor receptor by and The in to in clinically relevant cell and and and mesenchymal stem cell growth and in and used of the cells and in and BMP-2, BMP-6, and BMP-7 in cells and are was was to the the of and the the of in was in to the in was the of was to the of was the to the was of in a and was and by a and gene was a and gene to the used in this study are and of gene and receptor knockdown was the of to the by The of receptor in and was the the human of used in to and and to the relevant are in of to of to the and a and by of to The of of gene in was the and to of the used to the type and and the type and and to the of and a a the a and the of of was to and cells in to of gene of BMP-2, in in receptor and gene was by the of receptor and the of of of gene in the to the to the of receptor knockdown and the of and the of and The the by and of a of and was and the to gene was by and are in the and in and and and the of that the proteins the receptor by a the of the receptor and was the by the and used in the cells The cells a of of of the of receptor a of cells and in in to of and a was by to a of and was and The was the of the and proteins and the in to the by cells of the receptor and to of the was a by and the of to cells of the of the and of the receptor of the and of and and mesenchymal stem cell growth and in and used of the cells and in and BMP-2, BMP-6, and BMP-7 in cells and are was was to the the of and the the of in was in to the in was the of was to the of was the to the was of in a and was and by a and gene was a and gene to the used in this study are and The of gene and receptor knockdown was the of to the by The of receptor in and was the the human of used in to and and to the relevant are in of to of to the and a and by of to The of of gene in was the and to of the used to the type and and the type and and to the of and a a the a and the of of was to and cells in to of gene of BMP-2, in in receptor and gene was by the of receptor and the of of of gene in the to the to the of receptor knockdown and the of and the of and The the by and of a of and was and the to gene was by and are in the and in and and and the of that the proteins the receptor by a the of the receptor and was the by the and used in the cells The cells a of of of the of receptor a of cells and in in to of and a was by to a of and was and The was the of the and proteins and the in to the by cells of the receptor and to of the was a by and the of to cells of the of the and of the receptor of the and of BMP-4, BMP-6, and BMP-7 in of BMP-2, BMP-4, BMP-6, and BMP-7 to the of primary was the and The of was by the osteoinductive of the and was to the by of the of receptor and was of by The gene in the of in and that induce a gene in the of the to induce in by in in The was that induce of primary that the and of this of receptor utilization in was to the osteoinductive the of in and the of in primary The of receptor was and to was the of the and the type and type to and to and to and to of and was the of the to and the receptor in human the receptor was and to was the type receptor in the of and The in to in to to and to of type receptor and and of type was the type receptor in in to in and in to in The of in was the the was receptor a of of and a of in to human to of receptor the receptor of receptor the of and by of the to by to receptor to and in of type type knockdown was by the of knockdown and to in to and to and to in of was probably in the of by the of to the knockdown of the type that the a the of type type and type are by of and primary a of of of the of type type of the was by the of are to the of receptor in to by in the of receptor by the of the the of of cells the was in the the was the and to of of type type by BMP-2, BMP-4, BMP-6, BMP-7 was a primary The knockdown of the type and the of to induce the of and by that type type are to in primary in the of the in this the effects by and of the receptor knockdown the effects of receptor knockdown and that the of the of receptor and in by the and and primary a of of of the of type type cells BMP-2, BMP-4, BMP-6, BMP-7 and of was by and to are the of receptor knockdown to the the of that to the are receptor knockdown the and are by in type receptor utilization of by and BMP-7 by and by and The and was a by knockdown of in and of and BMP-4, and and The and was a by of knockdown was of the and and that and are the primary type the of in the of and a of and that and the to of type receptor to and of in of by BMPs. the of this was the was of and was to the and BMP-7 a by in a of by and a BMP-7 The BMP-7 in human bone by and by of knockdown was of the BMPs. The of and knockdown was and knockdown the and receptor knockdown of and a of of and in cells BMP-2, BMP-4, BMP-6, of and of was by and to are the of gene receptor knockdown to the the of the was in the the was the and BMP-7 of and and of and the knockdown of in of that the of by of the type receptor was by the that knockdown a in the knockdown of in a of and that the of by of the type receptor in this was by the that knockdown in a in by and that and BMP-7 type receptor in the of and of type receptor utilization by the and in primary and in by of and of and in by of of and of of and to to the of and and BMP-4, the knockdown of in of that the by in a of a was to by the knockdown of in of that the of by in of and BMP-4, a the of that a of and by of and The of the and that and by that a of and in a of and and to and to this the to and in a and to in to the and of and in cells proteins and was by proteins to the receptor in was by of and was of that and of the was The of the the to the that are that of the and the of a to and of and and cells in The cells of of the of receptor and and by a of to of by and the of to the of by of of of of of of of of of relevant receptor the of that the are in the this of of in the by that of that and of the was The of the in this study that was the to the of type and formation. of a of and of of to in the the in of of the the in the of and and in the of type of and BMP-2, BMP-4, BMP-6, and BMP-7 in of BMP-2, BMP-4, BMP-6, and BMP-7 to the of primary was the and The of was by the osteoinductive of the and was to the by of the of receptor and was of by The gene in the of in and that induce a gene in the of the to induce in by in in The was that induce of primary that the and of this of receptor utilization in was to the osteoinductive the of in and the of in primary The of receptor was and to was the of the and the type and type to and to and to and to of and was the of the to and the receptor in human the receptor was and to was the type receptor in the of and The in to in to to and to of type receptor and and of type was the type receptor in in to in and in to in The of in was the the was receptor a of of and a of in to human to of receptor the receptor of receptor the of and by of the to by to receptor to and in of type type knockdown was by the of knockdown and to in to and to and to in of was probably in the of by the of to the knockdown of the type that the a the of type and to of of type type by BMP-2, BMP-4, BMP-6, BMP-7 was a primary The knockdown of the type and the of to induce the of and by that type type are to in primary in the of the in this the effects by and of the receptor knockdown the effects of receptor knockdown and that the of the of receptor and in type receptor utilization of by and BMP-7 by and by and The and was a by knockdown of in and of and BMP-4, and and The and was a by of knockdown was of the and and that and are the primary type the of in the of and a of and that and the to The of type receptor to and of in of by BMPs. the of this was the was of and was to the and BMP-7 a by in a of by and a BMP-7 The BMP-7 in human bone by and by of knockdown was of the BMPs. The of and knockdown was and knockdown the and and BMP-7 of and and of and the knockdown of in of that the of by of the type receptor was by the that knockdown a in the knockdown of in a of and that the of by of the type receptor in this was by the that knockdown in a in by and that and BMP-7 type receptor in the of and of and to to the of and and BMP-4, the knockdown of in of that the by in a of a was to by the knockdown of in of that the of by in of and BMP-4, a the of that a of and by of and The of the and that and by that a of and in a of and and to and to this the to and in a and to in to the and of and in cells proteins and was by proteins to the receptor in was by of and was of that and of the was The of the the to the that are that of the and the of a to in the this of of in the by that of that and of the was The of the in this study that was the to the of type and formation. of a of and of of to in the the in of of the the in the of and and in the of type of and of this study was to evaluate receptor utilization the of in primary receptor knockdown revealed that the osteoinductive of in primary are the type and and the type and and to in this to the of of of the in cell a of in to of and and the in this cell type in cells growth of in primary human bone cells the and of receptor utilization in in in this study the of the of receptor members of the to the type was to to by was the type to osteoinductive in The of to a in of by are in the type receptor knockdown in receptor by and to of and and a of to and and was in cells of type by used to of and of the of cells of the receptor that a of receptor in cells to the of the to that a type of receptor to the in of the and a receptor of a of the type receptor and of the type and of the type receptor a that of a of by the type a in the of in cells and the to osteoinductive in primary members of the and to receptor and utilization of type in the of in a of The in are to induce of cells to BMP-7 effects of BMP-7 to of BMP-7 the BMP-6, was to transforming growth and the and of BMP-7 in the and of in a the of receptor receptor in human to the of in the of in the of receptor the that in receptor human in and in the in this study that BMP-2, BMP-4, BMP-6, and BMP-7 induce of primary in the to a of type type the effects of in receptor utilization and in the the in clinically relevant The of this study was to evaluate receptor utilization the of in primary receptor knockdown revealed that the osteoinductive of in primary are the type and and the type and and to in this to the of of The of the in cell a of in to of and and the in this cell type in cells growth of in primary human bone cells the and of receptor utilization in in in this study the of the of receptor members of the to the type was to to by was the type to osteoinductive in The of to a in of by are in the type receptor knockdown in receptor by and to of and and a of to and and was in cells of type by used to of and of the of cells of the receptor that a of receptor in cells to the of the to that a type of receptor to the in of the and a receptor of a of the type receptor and of the type and of the type receptor a that of a of by the type a in the of in cells and the to osteoinductive in primary members of the and to receptor and utilization of type in the of in a of The in are to induce of cells to BMP-7 effects of BMP-7 to of BMP-7 the BMP-6, was to transforming growth and the and of BMP-7 in the and of in a the of receptor receptor in human to the of in the of in the of receptor the that in receptor human in and in the in this study that BMP-2, BMP-4, BMP-6, and BMP-7 induce of primary in the to a of type type the effects of in receptor utilization and in the the in clinically relevant the receptor used in the and of of the knockdown