BMP-2에 의한 협부 지방 성체 줄기세포의 골형성
Osteogenesis by BMP-2 in adult stem cell derived from buccal fat pad
2
조회
0
다운로드
5
인용
0
참고문헌
키워드
초록
Bone morphogenetic proteins (BMPs) in combination with stem cells gain more significance for their use in bone tissue engineering. The mesenchymal stem cell can be differentiated into osteoblast by the treatment of BMP. The aim of this study is to characterize the osteogenic differentiation process of adult stem cells derived from buccal fat pad according to BMP-2 within culture media and decide the appropriate concentration of BMP-2 to facilitate osteogenesis. The authors procured the stem cell from buccal fat pad and analyzed for presence of stem cell by flow cytomety against CD-34, CD-105 and STRO-1. The buccal fat derived stem cells (BFDC) were treated by application of the different concentration with BMP-2 of 0, 10, 50, 100 and 200ng/ml, respectively. And their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase(ALP) staining, Alizarin red staining and RT-PCR for osteocalcin(OC) gene expression at 7, 14 and 21day of culture. Flow cytometric analysis and biochemical assays demonstrated that BFDC might be a distinguished stem cells, and mineralization was accompanied in proportion to BMP-2 concentration. However, with 100ng/ml concentration of BMP-2, the BFDC demonstrated most efficent staining pattern of ALP and Alizarin red. The feasibilty of the osteogenic differentiation in the group of both 50ng/ml and 200ng/ml of BMP-2 showed similar activity and relatively weaker than that of 100ng/ml. These results suggest that the BMP-2 stimulate osteogenesis by BFDC effectively and that bone induction might be controlled through negative regulatory feedback in higher concentration.
Abstract
Bone morphogenetic proteins (BMPs) in combination with stem cells gain more significance for their use in bone tissue engineering. The mesenchymal stem cell can be differentiated into osteoblast by the treatment of BMP. The aim of this study is to characterize the osteogenic differentiation process of adult stem cells derived from buccal fat pad according to BMP-2 within culture media and decide the appropriate concentration of BMP-2 to facilitate osteogenesis. The authors procured the stem cell from buccal fat pad and analyzed for presence of stem cell by flow cytomety against CD-34, CD-105 and STRO-1. The buccal fat derived stem cells (BFDC) were treated by application of the different concentration with BMP-2 of 0, 10, 50, 100 and 200ng/ml, respectively. And their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase(ALP) staining, Alizarin red staining and RT-PCR for osteocalcin(OC) gene expression at 7, 14 and 21day of culture. Flow cytometric analysis and biochemical assays demonstrated that BFDC might be a distinguished stem cells, and mineralization was accompanied in proportion to BMP-2 concentration. However, with 100ng/ml concentration of BMP-2, the BFDC demonstrated most efficent staining pattern of ALP and Alizarin red. The feasibilty of the osteogenic differentiation in the group of both 50ng/ml and 200ng/ml of BMP-2 showed similar activity and relatively weaker than that of 100ng/ml. These results suggest that the BMP-2 stimulate osteogenesis by BFDC effectively and that bone induction might be controlled through negative regulatory feedback in higher concentration.
참고문헌 (26)
- Urist MR, "Bone formation by autoinduction", Science, 150, pp.893~899 (1965)
- Celeste AJ, "Identification of transforming growth factor beta family members present in bone-inductive protein purified from bovine bone", Proc Natl Acad Sci USA, 87, pp.9843~9847 (1990)
- Sykaras N, "Effect of recombinant human bone morphogenetic protein-2 on the osseointegration of dental implants; a biomechanical study", Clin Oral Investig, 8, pp.196~205 (2004)
- Boyne PJ, "A feasibility study evaluating rhBMP-2", absorbable collagen sponge for maxillary sinus floor augmentation, 1997, pp.17, 11~25
- Sailer HF, "Application of purified bone morphogenetic protein (BMP) preparations in cranio-maxillo-facial surgery; Reconstruction in craniofacial malformations and post-traumatic or operative defects of the skull with lyophilized cartilage and BMP", J Craniomaxillofac Surg, 22, pp.191~199 (1994)
- Howell TH, "A feasibility study evaluating rhBMP-2", absorbable collagen sponge device for local alveolar ridge preservation or augmentation, 1997, pp.17, 124~139
- Wozney JM, "Novel regulators of bone formation: molecular clones and activities", Science, 242, pp.1528~1534 (1988)
- Pittenger MF, "Multilineage potential of adult human mesenchymal stem cells", Science, 284, pp.143~ (1999)
- Martin GJ, "Posterolateral intertransverse process spinal arthrodesis with rhBMP-2 in a nonhuman primate", J Spinal Disord, 12, pp.179~ (1999)
- Zuk PA, "Multi - lineage cells from human adipose tissue: implications for cell - based therapies", Tissue Eng, 7, pp.211~228 (2001)
- Zuk PA, "De Ugarte DA, Huang JI, Mizuno H, et al: Human adipose tissue is a source of multipotent stem cells", Mol Biol Cell, 13, pp.4279~4295 (2002)
- Halvorsen YC, "Adipose-derived stromal cell-their utility and potential in bone formation", Int J Obes Relat Metab Disord, 24, pp.41~44 (2000)
- 표성운, "협부지방에서 성체 줄기세포의 분리와 골모 세포로의 분화", 대한구강악안면외과학회지, 32(6), pp.524~529 (2006), 대한구강악안면외과학회
- M. Wan, "BMP signaling in skeletal development", Biochem Biophys Res Commun, 328, pp.651~657 (2005)
- K. Miyazono, "BMP receptor signaling; transcriptional targets, regulation of signals, and signaling crosstalk", Cytokine Growth Factor Rev, 16, pp.251~263 (2005)
- Reddi AH, "Role of morphogenetic proteins in skeletal tissue engineering and regeneration", Nat Biotechnol, 16, pp.247~252 (1998)
- Koch H, "Osteoblastic differentiation of human adult mesenchymal stem cells after through gene transfer of BMP-2 in the absence of dexamethasone", Z Orthop Ihre Grenzgeb, 143, pp.684~690 (2005)
- Murata M, "Bone induction in subcutaneous tissue in rats by a newly developed DNA-coated atelocollagen and bone morphogenetic protein", Br J Oral Maxillofac Surg, 40, pp.131~135 (2002)
- Li X, "Smad6 is induced by BMP-2 and modulates chondrocyte differentiation", J Orthop Res, 21, pp.908~913 (2003)
- Miyazono K, "Signal transduction by bone morphogenetic protein receptors: functional roles of Smad proteins", Bone, 25, pp.91~93 (1999)
- David H, "Bone morphogenetic proteins and spinal fusion", Neurosurg Focus, 13, pp.1~13 (2002)
- Zachos TA, "Gene-Mediated Osteogenic Differentiation of Stem Cells by Bone Morphogenetic Proteins-2 or -6", J Orthop Res, 24, pp.1279~1291 (2006)
- Bellows CG, "Determination of the capacity for proliferation and differentiation of osteoprogenitor cells in the presence and absence of dexamethasone", Dev Biol, 140, pp.132~138 (1990)
- Malaval L, "Cellular expression of bone-related proteins during in vitro osteogenesis in rat bone marrow stromal cell cultures", J Cell Physiol, 158, pp.555~572 (1994)
- Dragoo JL, "Tissue engineered bone from BMP-2-transduced stem cells derived from human fat", Plast Reconstr Surg, 115, pp.1665~ (2005)
- Knippenberg M, "Osteogenesis versus chondrogenesis by BMP-2 and BMP-7 in adipose stem cells", Biochem Biophys Res Comm, 342, pp.902~908 (2006)