• 생명과학회지
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• 제28권12호
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• pp.1448-1454
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• 2018

최근 들어 곤충을 식품 및 바이오 소재로 이용한 연구가 활발히 진행되고 있다. 그러나 곤충을 이용한 조골세포 활성 및 분화에 따른 골 형성 촉진 효과에 대한 연구는 아직 미흡한 실정이다. 뿐만 아니라 풀무치를 이용한 기능성 연구는 거의 이루어지지 않고 있다. 따라서 본 연구에서는 골 형성 촉진 효능을 가진 새로운 천연물 소재 개발을 위해 풀무치 추출물의 MG-63 조골세포의 분화 촉진 효과를 연구하였다. 조골세포에서 풀무치 추출물의 독성 및 증식 효과를 평가하기 위하여 MTS assay를 진행한 결과, $1,000{\mu}g/ml$ 농도까지 세포 독성이 나타나지 않았으며, 48시간 배양했을 때 $500-1,000{\mu}g/ml$ 농도에서 105%와 116%의 세포 증식 효능을 확인 하였다. 풀무치 추출물이 조골세포 분화에 미치는 영향을 확인하기 위하여 3일 및 5일간 풀무치 추출물을 MG-63 조골세포에 처리한 후 ALP 활성을 측정하였다. 그 결과 $100{\mu}g/ml$ 농도에서 positive control로 사용한 조골세포 분화배지(DM)군과 유사한 정도로 분화가 증가하였으며 500 및 $1,000{\mu}g/ml$ 농도에서는 2-3배까지 조골세포 분화가 촉진되었다. 이 결과는 ALP staining에서도 유사하게 나타났다. mRNA 발현량의 변화를 측정한 결과, Alpl과 Runx2 유전자 발현량이 증가하였고, 단백질 발현량을 측정했을 때에도 유사한 결과를 확인하였다. 이를 통해 ALP와 Runx2 유전자 및 단백질 발현에 의해서 ALP 활성이 증가하고 조골세포 분화가 촉진되었을 것으로 판단되며, 풀무치 추출물을 이용한 골 형성 촉진에 따른 골다공증 예방 및 치료 기능성 소재 개발에 대한 가능성을 확인하였다.

• 생명과학회지
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• 제29권9호
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• pp.1027-1033
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• 2019

현재 우리나라는 고령화 사회에 접어들었으며, 이와 함께 골 대사 질환이 큰 문제로 대두되고 있다. 그 중 가장 흔하게 발생하는 골다공증은 여러 요인에 의해 발생하고 있으며 이의 예방 및 치료를 위한 소재 개발 연구가 활발하게 진행되고 있다. 그러나 곤충을 소재로 한 골다공증 예방 및 치료 연구는 미흡한 실정이다. 따라서 본 연구에서는 골 형성 촉진 효능을 가진 새로운 소재 개발을 위해 갈색거저리 유충 오일의 MG-63 조골세포의 분화 촉진 효과를 연구하였다. 조골세포에서 갈색거저리 유충 오일의 독성 및 증식 효과를 평가하기 위하여 MTS assay를 진행한 결과 $80{\mu}g/ml$ 농도까지 세포 독성이 나타나지 않았으며, 48시간 배양했을 때 $40-80{\mu}g/ml$ 농도에서 120% 정도의 세포 증식 효능을 확인 하였다. 갈색거저리 유충 오일이 조골세포 분화에 미치는 영향을 확인하기 위하여 5일간 갈색거저리 유충 오일을 MG-63 조골세포에 처리한 후 ALP 활성을 측정하였다. 그 결과 $80{\mu}g/ml$ 농도에서 무처리 군에 비해 약 180%의 조골세포 분화 촉진 효능을 관찰 하였다. 이 결과는 ALP staining에서도 유사하게 나타났다. mRNA 발현량의 변화를 측정한 결과, Alpl과 Runx2 유전자 발현량이 증가하였고, 단백질 발현량을 측정했을 때에도 유사한 결과를 확인하였다. 이를 통해 ALP와 Runx2 유전자 및 단백질 발현에 의해서 ALP 활성이 증가하고 조골세포 분화가 촉진되었을 것으로 판단되며, 갈색거저리 유충 오일을 이용한 골 형성 촉진에 따른 골다공증 예방 및 치료 기능성 소재 개발에 대한 가능성을 확인하였다.

• Reproductive and Developmental Biology
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• 제37권4호
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• pp.225-232
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• 2013

Satellite cells were derived from muscular tissue in postnatal pig. Satellite cell is an important to growth and development in animal tissues or organs. However, the progress underlying induced differentiation is not clear. The aim of this study was to evaluate the morphologic and the transcriptome changes in porcine satellite cell (PSC) treated with insulin, rosiglitazone, or dexamethasone respectively. PSC was obtained from postnatal muscle tissue. In study 1, for study the effect of insulin and FBS on the differentiated satellite cells, cells were cultured at absence or presence of insulin treated with FBS. Total RNA was extracted for determining the expression levels of myogenic PAX3, PAX7, Myf5, MyoD, and myogenin genes by real-time PCR. Myogenic genes decreased expression levels of mRNA in treated with insulin. In study 2, in order to clarify the relationship between rosiglitazone and lipid in differentiated satellite cells, we further examined the effect of FBS on lipid accumulation in the presence or absence of the rosiglitazone and lipid. Significant differences were observed between rosiglitazone and lipid by FBS. The mRNA of FABP4 and $PPAR{\gamma}$ increased in rosiglitazone treatment. In study 3, we examined the effect of dexamethasone on osteogenic differentiation in PSC. The mRNA was increased osteoblasotgenic ALP and ON genes treated with dexamethasone in 2% FBS. Dexamethasone induces osteoblastogenesis in differentiated PSC. Taken together, in differentiated PSCs, FABP4 and $PPAR{\gamma}$ increased to rosiglitazone. Whereas, no differences to FBS and lipid. These results were not comparable with previous reports. Our results suggest that adipogenic, myogenic, and osteoblastogenic could be isolated from porcine skeletal muscle, and identify culture conditions which optimize proliferation and differentiation formation of PSC.

• Reproductive and Developmental Biology
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• 제37권4호
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• pp.219-224
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• 2013

Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90~100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson's, oil red O, and Alizarin red staining respectively. We performed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteoblast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were induced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strategies for augmenting meat quality.

• 셀메드
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• 제8권3호
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• pp.13.1-13.8
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• 2018

In the field of osteoporosis, there has been growing interest in anabolic agents that enhance bone formation. The purpose of this study was to examine the effects of NNMBS 246 osteoblastic differentiation with associated signaling pathways. NNMBS 246 markedly increased alkaline phosphatase (ALP) activity and calcium nodule formation. Stimulation with NNMBS 246 not only increased the differentiation markers (ALP, OPN, OCN) level and transcription markers (RUNX2, Osterix) mRNA expression but also upregulated the ECM molecules and OPG mRNA expression. Treatments of NNMBS 246 downregulated MMPs (MMP-1, MMP-2, MMP-9), but RANKL mRNA expression. Furthermore, NNMBS 246 activated osteoblastic differentiation markers and formed calcium nodules in human periodontal ligament cells (hPDLCs) and cementoblast cells. NNMBS 246 induced phosphorylation of MAPKs, Akt, nuclear p65 and IkB-${\alpha}$. BMP-2/Smad and ${\beta}$-catenin signaling pathways were activated by NNMBS 246. Sirtinol (SIRT1 inhibitor) inhibited NNMBS 246-induced osteoblastic differentiation markers mRNA expression. These results suggested that NNMBS 246 has the potential to enhance osteoblastogenesis probably through the activation of BMP/Smad and ${\beta}$-catenin signal pathways, and SIRT1 plays as critical mediator in bone anabolic effect of NNMBS 246.

• 대한한방부인과학회지
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• 제27권2호
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• pp.23-33
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• 2014

Objectives: In this study, the author aimed to evaluate the effect of EtOH extract of Ganoderma lucidum (GLE) on osteoblast proliferation in rat fetus calvarial cells. Methods: The osteoblast separated from rat fetus calvariae was cultivated for 6~21 days and evaluated the cell function. After the addition of GLE on the culture medium, we determined the effect of GLE on the cell viability, cell proliferation, bone matrix protein synthesis, alkaline phosphatase (ALP) activity, collagen synthesis and calcified nodule formation of the cultivated osteoblast. Results: GLE did not change the survival rate of rat calvarial osteoblast. GLE increased the proliferation of rat calvarial osteoblast. GLE increased ALP activity of rat calvarial osteoblast. GLE increased bone matrix protein synthesis of rat calvarial osteoblast. GLE increased collagen synthesis of rat calvarial osteoblast. GLE slightly affected calcified nodule formation of rat calvarial osteoblast. Conclusions: This study suggests that Ganoderma lucidum might improve the osteoporosis resulted from augmentation of osteoblast proliferation.

• 대한한방부인과학회지
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• 제29권2호
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• pp.1-14
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• 2016

Objectives : In this study, the author aimed to evaluate the effects of water extract of Dokwhalgisaeng-tang Gamibang (DGG) on osteoblast proliferation in murine calvarial cells. Methods : The osteoblast separated from murine calvariae was cultivated and evaluated the cell function. After the addition of DGG on the culture medium, we determined the effect of DGG on the cell proliferation, protein synthesis, alkaline phosphatase activity, collagen synthesis, and cell viability of the cultivated osteoblast in the presence of dexamethasone. Results : DGG increased the survival rate, proliferation, alkaline phosphatase (ALP) activity, protein synthesis and collagen synthesis of rat calvarial osteoblast in the presence of dexamethasone. Conclusions : DGG might improve the osteoporosis resulted from augmentation of osteoblast proliferation.

• Journal of Microbiology and Biotechnology
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• 제28권10호
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• pp.1614-1625
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• 2018

Periodontitis, which is a severe inflammatory disease caused by endotoxins secreted from oral pathogens, destructs gingival tissue and alveolar bone. Curcuma xanthorrhiza, commonly called Java turmeric, has been shown to possess anti-bacterial and anti-inflammatory activities. The present study evaluated the inhibitory effect of C. xanthorrhiza supercritical extract (CXS) standardized with xanthorrhizol on lipopolysaccharide (LPS)-induced periodontitis in an animal model. LPS was topically injected into the periodontium of Sprague-Dawley rats to induce periodontitis and CXS (30 and $100mg{\cdot}kg^{-1}{\cdot}day^{-1}$) was orally administered after day 12. Histologically, CXS inhibited the collapse of gingival tissue by preventing cell infiltration. CXS significantly downregulated the expression of matrix metalloproteases (MMPs) and inflammation-related biomarkers, such as nuclear factor-kappa B ($NF-{\kappa}B$) and interleukin-1 beta ($IL-1{\beta}$) in gingival tissue. CXS also improved bone remodeling by downregulating osteoclastic transcription factors, such as nuclear factor of activated T-cells c1 (NFATc1), tartrate-resistant acid phosphatase (TRAP), and cathepsin K. In addition, CXS upregulated osteoblast differentiation-related markers, alkaline phosphate (ALP) and collagen type I alpha (COLA1). Thus, CXS can ameliorate periodontitis by inhibiting inflammation and improving bone remodeling.

• 대한한방부인과학회지
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• 제26권3호
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• pp.33-43
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• 2013

Objectives: Osteoporosis is characterized by bone loss and morbidity with osteoporotic fracture. In this study, the author aimed to evaluate the effect of dried roots of Rehmannia glutinosa extract (RGE) on osteoblast proliferation in murine calvarial cells. Methods: The osteoblast separated from murine calvariae was cultivated for 6 days and evaluated the cell function. After the addition of RGE on the culture medium, we determined the effect of RGE on the cell viability, cell proliferation, protein synthesis, alkaline phosphatase activity, collagen synthesis and calcified nodule formation of the cultivated osteoblast. Results: The results were summarized as follows. 1. RGE did not change the survival rate of rat calvarial osteoblast. 2. RGE increased the proliferation of rat calvarial osteoblast. 3. RGE increased ALP activity of rat calvarial osteoblast., 4. RGE slightly affected protein synthesis of rat calvarial osteoblast. 5. RGE increased collagen synthesis of rat calvarial osteoblast. 6. RGE slightly affected calcified nodule formation of rat calvarial osteoblast. Conclusions: From these results, it is concluded that RG might improve the osteoporosis resulted from augmentation of osteoblast proliferation.

• The Korean Journal of Physiology and Pharmacology
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• 제23권1호
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• pp.47-54
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• 2019

Estrogen withdrawal in post-menopausal women leads to overactivation of osteoclasts, which contributes to the development of osteoporosis. Inflammatory cytokines are known as one of mechanisms of osteoclast activation after estrogen deficiency. SPA0355 is a thiourea derivative that has been investigated for its antioxidant and anti-inflammatory activities. However, its efficacy in bone resorption has not been previously investigated. The aim of this study was to investigate the impact of SPA0355 on the development of osteoporosis and to explore its mode of action. In vitro experiments showed that SPA0355 inhibited receptor activator of $NF-{\kappa}B$ ligand (RANKL)-induced osteoclastogenesis in primary bone marrow-derived macrophages. This effect appears to be independent of estrogen receptor activation as ICI 180,782 failed to abrogate its effects on osteoclasts. Further signaling studies revealed that SPA0355 suppressed activation of the MAPKs, Akt, and $NF-{\kappa}B$ pathways. SPA0355 also increased osteoblastic differentiation, as evidenced by its effects on alkaline phosphatase activity and mineralization nodule formation. Intraperitoneal administration of SPA0355 to ovariectomized mice prevented bone loss, as verified by three-dimensional images and bone morphometric parameters derived from ${\mu}CT$ analysis. Noticeably, SPA0355 did not show hepatotoxicity and nephrotoxicity and also had little effect on hematological parameters. Taken together, the results indicate that SPA0355 may protect against bone loss in ovariectomized mice by stimulation of osteoblast differentiation and by inhibition of osteoclast resorption. Therefore, SPA0355 is a safe and potential candidate for management of postmenopausal osteoporosis.