척추 황색 인대 세포에서 Bone Morphogenetic Protein-2 cDNA 전달에 의한 골형성 유도 : 조직 공학적 접근의 가능성

문성환,김향,권언혜,원정훈,김학선,한수봉,이환모 대한척추외과학회 2002 대한척추외과학회지 Vol.9 No.4

두개봉합부의 초기형태발생과정에서 BMP와 그 수용체의 발현 양상

윤양하,이상원,박미현,류현모,남순현,김영진,김현정,Yune, Yang-Ha,Lee, Sang-Won,Park, Mi-Hyun,Ryoo, Hyun-Mo,Nam, Soon-Hyeun,Kim, Young-Jin,Kim, Hyun-Jung 대한소아치과학회 2002 大韓小兒齒科學會誌 Vol.29 No.3

Bone morphogenetic proteins(BMPs)는 형태형성 및 세포 분화동안 다양한 조절 역할을 담당하는 신호전달 인자이다. 시상두개봉합부 발생시 BMPs와 그 수용체의 역할을 분석하기 위해, in situ hybridization방법을 이용하여 태생 15일에서 18일 시상두개봉합부에서 그 발현 양상을 분석하였다. BMP-2와 BMP-3은 태생 15일부터 osteogenic front와 두정골에서 발현을 보였으며 태생 16일부터 모낭에서 발현이 시작되었다. BMP-4는 osteogenic front에서 강하게 발현되었으며, 간엽조직 및 두정골에서 약하게 발현되었다. BMP-5는 모낭에서 발현되었다. 이전 연구에서 BMP-6는 비후된 연골세포에서 발현된다고 보고되었으나 본 연구에서는 발현되지 않았다. BMP-7은 태생기에 두정골에서 발현되었다. BMPR-IB는 osteogenic front의 외측 가장자리에서 발현되었으나, BMPR-IA는 발현되지 않았다. 이런 결과를 종합해 볼 때, 두개봉합부 초기 형태발생시 BMP-4는 미분화 간엽세포로부터 골아세포로 commit되는 초기단계에 중요한 역할을 하며, BMP-2와 BMP-3는 전구 골아세포에서 골아세포로의 분화과정에, BMP-7은 좀 더 분화가 진행된 골아세포 및 골의 분화 유지에 중요하며, type I 수용체 중 BMPR-IB가 BMP들의 신호전달에 중요한 역할을 함을 예측 할 수 있다. 결론적으로 BMP 신호전달은 다양한 BMP 리간드들과 그 수용체들에 의해 골아세포 분화 전반에 걸쳐 관여하고 있음을 시사한다. Bone morphogenetic proteins(BMPs) are secretory signal molecules which have a variety of regulatory functions during morphogenesis and cell differentiation. To evaluate roles of BMPs and their receptors on mouse sagittal suture development, we have examined their expression patterns in serial sections of sagittal sutures by in situ hybridization during embryonic stages(E15-E18). BMP-2 and BMP-3 were expressed in the osteogenic front and parietal bone on embryonic 15day, from E16 in hair follicle. BMP-4 was strongly expressed in the osteogenic front and weakly expressed in the mesenchyme and parietal bone. BMP-S was expressed in the hair follicles. BMP-6 was not expressed in this study. BMP-7 was expressed in parietal bone during embryonic stage. BMPR-IB was expressed in the osteogenic front, but BMPR-IA was not. From these datas, we suggest that the BMP-4 regulates the early commitment of mesenchymal cells to the osteogenic lineages, the BMP-2 and BMP-3 may be involved in regulating the differentiation of osteoblast precursor cells. BMP-7 was involved in maintenance of differentiated osteoblasts. BMPs were key signaling molecules that regulate early calvarial bone morphogenesis, mediated by BMPR-IB.

Reports : IGF1 potentiates BMP9-induced osteogenic differentiation in mesenchymal stem cells through the enhancement of BMP/Smad signaling

( Liang Chen ),( Xiang Zou ),( Ran-xi Zhang ),( Chang-jun Pi ),( Nian Wu ),( Liang-jun Yin ),( Zhong-liang Deng ) 생화학분자생물학회(구 한국생화학분자생물학회) 2016 BMB Reports Vol.49 No.2

Engineered bone tissue is thought to be the ideal alternative for bone grafts in the treatment of related bone diseases. BMP9 has been demonstrated as one of the most osteogenic factors, and enhancement of BMP9-induced osteogenesis will greatly accelerate the development of bone tissue engineering. Here, we investigated the effect of insulin-like growth factor 1 (IGF1) on BMP9-induced osteogenic differentiation, and unveiled a possible molecular mechanism underling this process. We found that IGF1 and BMP9 are both detectable in mesenchymal stem cells (MSCs). Exogenous expression of IGF1 potentiates BMP9-induced alkaline phosphatase (ALP), matrix mineralization, and ectopic bone formation. Similarly, IGF1 enhances BMP9-induced endochondral ossification. Mechanistically, we found that IGF1 increases BMP9-induced activation of BMP/Smad signaling in MSCs. Our findings demonstrate that IGF1 can enhance BMP9-induced osteogenic differentiation in MSCs, and that this effect may be mediated by the enhancement of the BMP/Smad signaling transduction triggered by BMP9. [BMB Reports 2016; 49(2): 122-127]

생쥐의 oviduct에서 estrogen에 의한 bone morphogenetic protein 2, 4 (BMP2, BMP4)의 발현 조절 및 기능

조남희,계명찬 한국발생생물학회 2017 한국발생생물학회 학술발표대회 Vol.2017 No.8

골 형성 단백질(Bone morphogenetic protein, BMP)은 TGF-β superfamily의 구성원 중에 하나이며, 이들은 원래 뼈 형성을 유도하는 능력에 의해 발견되었지만, 뼈 외에도 다른 세포 및 기관의 성장과 분화를 조절하는 것으로 밝혀졌다. 다기능적인 골 형성 단백질은 포유동물의 reproduction에도 매우 중요한 역할을 하는데, 예를 들어 BMP8A와 BMP8B는 생쥐의 정자 형성과 부고환에 일정한 기능을 하는 것으로 보고되었고 난소에서의 BMP15는 난포성장을 자극하고, 과립막세포(granulosa cell)의 증식시키는데 관여하는 단백질로 알려져 있다. 하지만 난관(oviduct)에서의 BMP 역할은 알려진 바가 적다. 그래서 본 연구에서는 골 형성 단백질이 착상 전 oviductal environment에 어떠한 영향을 미치는지 밝히고자 하였다. 8주령 생쥐의 Estrous cyclic oviduct을 가지고 RT-PCR과 Immunohistochemistry(IHC)를 통해 BMP2, 4의 mRNA와 단백질 발현을 확인하였다. Estrogen에 의한 BMP2, 4의 영향을 확인하고자 난소절제술을 시행한 생쥐와 Estrogen receptor alpha(ERα) Knockout 생쥐를 통해 mRNA와 단백질 발현을 확인하였다. Oviduct의 ciliated cell을 가지고 BMP2, 4의 기능을 밝히고자 siRNA실험을 진행 하였다. Estrous cyclic oviduct cDNA를 통해 RT-PCR한 결과, 이 중 Estrus 시기에 가장 높은 발현을 보인 BMP2, 4의 mRNA level은 Isthmus보다 Infundibulum과 Ampulla에 증가하였고, Immunohistochemistry (IHC)를 통해 BMP2와 BMP4 단백질은 난관 상피세포 중에서 ciliated cell에 발현되었다. 이를 ciliated cell marker인 β-tubulin과 함께 Immunofluorescence(IF)를 진행한 결과, 주로 β-tubulin-positive ciliated cell에서 발현됨을 확인하였다. 난소 절제술을 시행한 생쥐의 난관에서는 E2와 DPN (ERβ agonist)에 의해 BMP2, 4 mRNA 발현이 증가하고, ERαKO 난관의 경우 WT(Die)에 비해 BMP2, 4 mRNA, 단백질의 발현 모두 증가하였다. BMP2와 BMP4의 기능을 밝히고자 Ciliated cell line인 OA-6b를 가지고 siRNA 실험을 진행한 결과, Bmp2, Bmp4 siRNA 처리군에서 Ciliated cell marker인 FOXJ1의 발현이 줄어들고 Proliferation marker인 Ki67, Pcna의 발현이 낮아졌다. 이로써, Oviduct 내 BMP2와 BMP4 발현은 Estrogen과 양의 상관성이 있으며, Ciliated cell에서 Estrogen - ER β signaling을 통해 조절된다. Estrogen에 의해 유도되는 BMP2와 BMP4는 ciliated cell에서 Autocrine, Paracrine factor로 작용할 가능성이 있다. 더 나아가 Oviduct의 Infundibulum 및 Ampulla에 강하게 발현되는 BMP2, BMP4는 난자 및 배아에 autocrine, paracrine factor로 작용하여 oviductal cell proliferation을 조절하고, 수정을 위한 oviductal environment 조성에 중요한 역할을 하는 것으로 사료된다.

Smad에 의한 alkaline phosphatase 유전자의 발현 조절기전

김난진,류현모,김현정,김영진,남순현 대한소아치과학회 2001 大韓小兒齒科學會誌 Vol.28 No.2

본 실험은 탁월한 골유도능으로 관심의 대상이 되고 있는 RMP의 세포내 신호 전달자로 알려진 Smad 1과 Smad 5가 조 골세포 초기 분화표지인자인 ALP 유전자의 발현에 미치는 영향 및 그 조절기전을 알아보고자 하였다. BMP 처리 없이도 Smad에 의해 ALP가 발현되는가를 알아보기 위해 Smad 1과 Smad 5가 각각 stably transfection된 C2C12 세포를 3일간 배양후 histochemical assay를 하였고, Smad 1과 Smad 5의 expression vector와 ALP promoter vector를 transient co-transfectiongksgn ALP promoter activity를 측정하였다. Smad에 의한 BMP의 효과를 알아보기 위해서 100ng/ml의 BMP-2를 처리한 군과 처리하지 않은 군으로 나누어 세포를 배양한후 ALP 유전자의 발현을 northern blot analysis로 확인하였다. Smad가 ALP 유전자의 발현을 직접적으로 조절하는가를 알아보기 위여서는 단백질 합성억제제인 cycloheximide를 전처리하여 ALP 유전자의 발현을 northern blot analysis하였다. 이상의 실험결과 다음과 같은 결론을 얻었다. ·Smad 1과 Smad 5가 과발현된 세포에서는 BMP 처리없이도 ALP가 발현된다. ·Smad 1과 Smad 5가 과발현된 세포에서 RMP 처리후 ALP 발현 증가율이 대조군 보다 현저히 높게 나타나 Smad가 BMP 효과를 증가시킨다는 것을 알수있다. ·Smad는 새로운 단백질의 합성을 통해 ALP 유전자를 발현시킨다. Bone morphogenetic proteins(BMPs), members of the transforming growth factor β(TGF-β) superfamily were first identified as the factors that induce ectopic bone formation in vivo, when implanted into muscular tissue. Especially BMP-2 inhibits terminal differentiation of C2C12 myoblasts and converts them into osteoblast lineage cells. In the molecular mechanism of the signal transduction of TGF-β and related factors, intracellular signaling proteins were identified as Smad. In previous study, it has been reported that Smad 1 and Smad 5, which belong to the R-Smad family mediate BMP signaling, were involved in the induction of osteoblast differentiation in C2C12 cells. To understnad the role of Smads involved in osteogenic transdifferentiation in C2C12 cell, in present study, after we stably transfected C2C12 cells with each. Smad(Smad 1, Smad 5) expression vector, cultured for 3 days and stained for alkaline phophatase activity. ALP activity positive cells appeared in the Smad 1, Smad 5 stably transfected cell even in the abscence of BMP. After transiently co-transfected C2C12 cells with each Smad expression vector and ALP promoter, it was excence of MBP. These result suggested that both Smad 1 and Smad 5 were involved in the intracellular BMP signals which induce osteoblast differentiation in C2C12 cells. The effect of BMP on C2C12 cells with Smad 1, Smad 5 transfected were studied by using northern blot analysis. the treatment of BMP upregulated ALP mRNA level in three groups, especially upregulation of ALP was larger in Smad 1, Smad 5 transfected cell than control group. Pretreatment with cycloheximide(10㎍/ml), a protein synthesis inhibitor resulted in blocking the ALP gene expression even in BMP(100ng/ml) treated cell. These results suggested that Smad increased the level of ALP mRNA via the synthesis of a certain transcriptional regulatory protein.

아데노바이러스 벡터를 이용한 BMP-2와 FGF-2의 동시 발현이 골형성에 미치는 영향

송상훈,장원구,김선헌,국민석,김옥준,김인애,박인규,고정태 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.13

FGF-2 and BMPs are expressed simultaneously during the embryonic bone development and repair of fractured bone. Delivery of BMP-2 peptide and gene has been evaluated for a therapeutic bone regeneration. However, low efficacy and short half -life of BMP-2 peptide have limited their clinical uses. This study was to determine the effects of BMP-2 and FGF-2 combination on bone formation with the adenoviral BMP-2 and FGF-2 gene delivery. Adenoviruses (Ad) expressing BMP-2 or FGF-2 were constructed under the CMV promoter, and characterized. AdBMP-2 dose-dependantly induced BMP-2 peptide expression in BLK cells, and increased alkaline phosphatase (ALP) activity in C2C12. AdFGF-2 also induced FGF-2 peptide expression in a dose-dependent manner. Proliferation of C3H10T1/2, ST-2 and MC3T3-E1 cell was stimulated by AdFGF-2 alone treatment, but ALP activity of the cells was inhibited. To evaluate the effect of BMP-2 and FGF-2 combination on bone formation, AdBMP-2 (100 moi) and/or AdFGF-2 (100 or 200 moi) were transduced into BLK fibroblast cells, and then the cells were transferred into subcutaneous spaces with collagen hydrogel in mice. Microradiographic analysis, biochemical assays and histology showed AdBMP-2 and AdFGF-2 combination produced less bone than AdBMP-2 alone at 2 weeks after implantation. In addition, AdBMP-2/AdFGF-2 combination group had less ALP activity and concentrations of inorganic phosphate and calcium in implants than AdBMP-2 treated group. These results suggest that FGF-2 may stimulate the proliferation of osteoblastic cells, but inhibit the BMP-2 induction of osteoblast differentiation and bone formation. FGF-2 and BMPs are expressed simultaneously during the embryonic bone development and repair of fractured bone. Delivery of BMP-2 peptide and gene has been evaluated for a therapeutic bone regeneration. However, low efficacy and short half -life of BMP-2 peptide have limited their clinical uses. This study was to determine the effects of BMP-2 and FGF-2 combination on bone formation with the adenoviral BMP-2 and FGF-2 gene delivery. Adenoviruses (Ad) expressing BMP-2 or FGF-2 were constructed under the CMV promoter, and characterized. AdBMP-2 dose-dependantly induced BMP-2 peptide expression in BLK cells, and increased alkaline phosphatase (ALP) activity in C2C12. AdFGF-2 also induced FGF-2 peptide expression in a dose-dependent manner. Proliferation of C3H10T1/2, ST-2 and MC3T3-E1 cell was stimulated by AdFGF-2 alone treatment, but ALP activity of the cells was inhibited. To evaluate the effect of BMP-2 and FGF-2 combination on bone formation, AdBMP-2 (100 moi) and/or AdFGF-2 (100 or 200 moi) were transduced into BLK fibroblast cells, and then the cells were transferred into subcutaneous spaces with collagen hydrogel in mice. Microradiographic analysis, biochemical assays and histology showed AdBMP-2 and AdFGF-2 combination produced less bone than AdBMP-2 alone at 2 weeks after implantation. In addition, AdBMP-2/AdFGF-2 combination group had less ALP activity and concentrations of inorganic phosphate and calcium in implants than AdBMP-2 treated group. These results suggest that FGF-2 may stimulate the proliferation of osteoblastic cells, but inhibit the BMP-2 induction of osteoblast differentiation and bone formation.

Polyethyleneimine-응축 BMP-2 발현 유전자를 이용한 골형성 효과

정희선,김경화,박윤정,김태일,이용무,구영,류인철,이동수,이승진,정종평,한수부,설양조,Cheong, Hee-Sun,Kim, Kyoung-Hwa,Park, Yoon-Jeong,Kim, Tae-Il,Lee, Yong-Moo,Ku, Young,Rhyu, In-Chul,Lee, Dong-Soo,Lee, Seung-Jin,Chung, Chong-Pyoung,Han, 대한치주과학회 2007 Journal of Periodontal & Implant Science Vol.37 No.4

Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.

Sustained BMP-2 delivery and injectable bone regeneration using thermosensitive polymeric nanoparticle hydrogel bearing dual interactions with BMP-2

Seo, B.B.,Choi, H.,Koh, J.T.,Song, S.C. Elsevier Science Publishers 2015 Journal of controlled release Vol.209 No.-

Localized and continuous osteogenic stimulation to defected sites is required for effective bone regeneration. Here, we suggest an injectable and sustained bone morphogenetic protein-2 (BMP-2) release system using thermosensitive polymeric nanoparticles bearing dual interacting forces with BMP-2. For sustained BMP-2 release, hydrophobic and ionic interactions were introduced to thermosensitive poly(phosphazene). Hydrophobic isoleucine ethyl ester and hydrophilic poly-ethylene glycol were mainly substituted to the poly(phosphazene) back bone for amphiphilicity and hydrophobic interaction with BMP-2. Carboxylic acid moiety was additionally substituted to the back bone for ionic interaction with BMP-2. These dual interacting polymeric nanoparticles (D-NPs) formed compact nanocomplexes with BMP-2. The aqueous solution of BMP-2/D-NP nanocomplexes was transformed to hydrogel when the temperature of the solution increased. Loaded BMP-2 was sustain-released for three weeks from the BMP-2/D-NP nanocomplex hydrogel. The extended BMP-2 exposure caused higher osteocalcin secretion in C2C12 cells. Significant bone generations were observed at the target site by single injection of BMP-2/D-NP nanocomplexes in vivo.

Apoptosis of oral epithelial cells in oral lichen planus caused by upregulation of BMP-4

Kim, S.-G.,Chae, C.-H.,Cho, B.-O.,Kim, H.-N.,Kim, H.-J.,Kim, I.-S.,Choi, J.-Y. Munksgaard International Publishers 2006 Journal of oral pathology & medicine Vol.35 No.1

<P>Background: </P><P>Bone morphogenic protein (BMP-4) is a member of transforming growth factor (TGF-&bgr;) family and involved in various functions including apoptosis during neural ectoderm development. The objective of this study is to determine whether BMP-4 is involved in apoptosis, one characteristic, of human oral lichen planus (OLP).</P><P>Methods: </P><P>Immunohistochemistry and <I>in situ</I> hybridization for BMP-4 were carried out in OLP (<I>n</I> = 21) and normal human oral mucosa (NOM, <I>n</I> = 31). Five tissue samples from NOM and OLP were underwent reverse transcriptase-polymerase chain reaction (RT-PCR). <I>In vitro</I> organ culture of oral mucosa was carried out with beads soaked with various concentration of BMP-4 (0.1, 1, and 10 <I>μ</I>g/ml). The samples from <I>in vitro</I> organ culture were undergone haematoxylin and eosin staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labelling technique (TUNEL) assay, and immunohistochemical study with p53, matrix metalloproteinases (MMP)-1, and MMP-3. Involucrin expression was determined by western blot analysis after treatment with BMP-4 or TGF-<I>&bgr;</I>1 on human oral keratinocytes.</P><P>Results: </P><P>In immunohistochemical analysis, expression of BMP-4 was higher in OLP than NOM. BMP-4 mRNA expression was also detected in epithelial cells of both NOM and OLP together with underlying T-lymphocytes by <I>in situ</I> hybridization and RT-PCR. In oral mucosa organ culture, BMP-4 soaked beads induced apoptosis of epithelial cells. Acantolysis combined with apoptosis in oral epithelium was observed at 1 <I>μ</I>g/ml of BMP-4 beads and it was due in part to the induction of p53 and MMP-1. Even MMP-3 induction was found in lower concentration of BMP-4 (0.1 and 1 <I>μ</I>g/ml). Moreover, the expression of MMP-1 and MMP-3 was also observed in OLP. Recombinant BMP-4 or TGF-<I>&bgr;</I>1 increased involucrin expression in human oral keratinocytes cell line.</P><P>Conclusions: </P><P>Expression of BMP-4 of epithelial cells was higher in OLP than NOM. High concentration of BMP-4 caused an apoptosis of oral epithelial cells in oral mucosa organ culture. Therefore, over-expression of BMP-4 is one causing factor for apoptosis of oral epithelial cells through upregulation of p53, MMP1 and MMP3 in OLP.</P>

가토 상악동 점막 거상 후 DBBP를 이식재로 사용시 BMP4, BMP6의 발현

이현석,허현아,표성운,이원,Lee, Hyun-Suk,Heo, Hyun-A,Pyo, Sung-Woon,Lee, Won 대한악안면성형재건외과학회 2007 Maxillofacial Plastic Reconstructive Surgery Vol.29 No.6

이 실험은 이식재로 Deproteinated Bovine Bone Powder (DBBP)를 사용하였을 때와 골 이식재를 사용하지 않고 단순히 Absorbable Gelatin Sponge(AGS)만을 사용하였을 때의 BMP4와 BMP6의 상대적인 발현량을 real-time RT-PCR을 이용하여 비교해 보기 위한 것으로 다음과 같은 결과를 얻었다. 1. BMP4의 경우 처음 1일째와 3일째의 경우 DBBP군과 AGS군 모두 일정하게 증가하였으나 5일째 AGS군에서 감소하다가 7일째 다시 증가하였으며 9일째 다시 감소하였다. DBBP군에서는 7일까지 계속하여 증가하다가 9일째 감소하였다. DBBP군이 AGS군에 비해 발현의 양이 많은 경우가 많았지만 유의성은 없었다 (p>0.05). 2 BMP6의 경우 처음 1일째와 3일째의 경우 DBBP군과 AGS군 모두 일정하게 증가하였으나 5일째 AGS군에서 감소하다가 7일째 다시 증가하였으며 9일째 다시 감소하였다. DBBP군에서는 7일까지 계속하여 증가하다가 9일째 감소하였다. AGS군이 DBBP군에 비해 발현의 양이 많은 경우가 많았지만 유의성은 없었다 (p>0.05). 3. 두 군간에 동일시기에 BMP발현이 유의할 만한 차이를 보이지 않는 것은 DBBP와 AGS 모두 space retainer로서 작용을 하여 혈병중의 BMP발현의 양상이 비슷하기 때문으로 여겨진다. 4. 따라서 DBBP가 AGS에 비해 초기 골재생에 크게 유리한 점은 없는 것으로 여겨지며 초기 골 형성에서 BMP의 발현은 이식재의 종류가 아니라 물리적인 carrier로서의 작용이 더 중요한 것으로 여겨진다. The most important factor for successful implantation is osseointegration between the implant and bone. The expression of bone morphogenetic proteins (BMPs) inducing bone formation would differ after maxillary sinus elevation. And within the same graft material. the expression of BMPs would change with time after graft. The aim of this study was to compare the relative expressions of BMP4 and BMP6 using real-time RT-PCR when maxillary sinus elevation was performed using deproteinated bovine bone powder (DBBP) as the graft material or absorbable gelatin sponge (AGS) as the filler without any graft material. Fifteen rabbits, each weighing between 3.0 to 3.5 Kg, were divided randomly into 5 groups of 3 animals each based on their time of sacrifice 0, 3, 5, 7 and 9 days). After exposure of the maxillary sinus bilaterally, bone graft was performed in the right maxillary sinus using DBBP ($BBP^{(R)}$ Oct Inc., Cheonan, Korea) and only AGS ($Gelfoam^{(R)}$ Pharmacia & Upjohn Company, Kalamazoo, MI, U.S.A) was placed into the left without any graft material. Each group of rabbits was sacrificed at 1, 3, 5, 7, or 9 days after operation and all specimens were harvested. And the following results were obtained using real-time RT-PCR from isolated total RNA of the samples. 1. The expression of BMP4 increased at postoperative 1 and 3 days in both DBBP group and AGS group. In AGS group. it decreased at postoperative 5 days. increased again at postoperative 7 days, and decreased at postoperative 9 days. In DBBP group, it increased until postoperative 7 days and decreased at postoperative 9 days. Although the expression of BMP4 was higher in DBBP group compared with AGS group, it was not statistically significant (p>0.05). 2. The expression of BMP6 increased at postoperative 1 and 3 days in both DBBP group and AGS group. In AGS group, it decreased at postoperative 5 days, increased again at postoperative 7 days, and decreased at postoperative 9 days. In DBBP group, it increased until postoperative 7 days and decreased at postoperative 9 days. Although the expression of BMP6 was higher in AGS group compared with DBBP group, it was not statistically significant (p>0.05). 3. There was no statistically significant difference in BMP expression in both groups during same period of time. It' s probably because DBBP and AGS both functioned as a space retainer so that the BMP expression in blood clot seemed to be similar. 4. Thus, DBBP would not offer many benefits for early bone regeneration compared with AGS. The expression of BMP in early bone formation seems to be more influenced by physical carrier rather than the graft type.