• 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.

• Journal of Periodontal and Implant Science
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• 제28권2호
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• pp.235-247
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• 1998

Calcium sulfate has a long history of medical use as an implant material. The biocompatibiliry of the material has been clearly established. Bone ingrowth concomitant with resorption occurs rapidly with efficient conduction of bone from particle to particle. Calcium sulfate also has a potential for functioning as a good bamer membrane. The purpose of this study was to compare the biocompatibility of different types of calcium sulfate grafting materials including an expelimental calcium sulfate compound on periodontal ligament cells in vitro as a preliminary test towards the development of a more convenient and useful form of grafting material which could promote regeneration of periodontal tissue. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. cells were cultured in a.MEM culture medium containing 20% FBS, at $37^{\circ}C$ and 100% humidity, in a 5% CO2 incubator. Cells were cultured into 96 well culture plate $1{\times}104$ cells per well with $\alpha$-MEM and incubated for 24 hours. After discarding the medium, those cells were cultured in $\alpha$-MEM contained with 10% FBS alone (control group), in medcal-grade calcium sulfate(MGCS group), in plaster(plaster group), experimental calcium sulfate paste(CS paste group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTI assay, collagen synthesis. The results \vere as follows. 1. In the analysis of cell proliferation by cell counting, both medical-grdde calcium sulfate group and plaster group showed no stastically significant difference at day 1, 2, 3 accept for plaster group at day 1 compared to control group, but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.05). 2. In the analysis of cytotoxicity by MIT assay, both medical-grade calcium sJlfate group and plaster group showed no stastically significant difference compared to control group at day 1, 2, 3 but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.OS). 3. In the analysis of collagen synthesis by immunoblotting assay, high level was detected for medical-grade calcium sulfate group and plaster group at day 1, 2, 3 compared to CS paste group. On the basis of these results, medical-grade calcium sulfate and plaster was shown to possess biocompatibility whereas the CS paste had unfavourable outcome. This observation shows a need for modification of the materials contained in calcium sulfate paste.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제34권2호
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• pp.155-161
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• 2012

Platelet-rich fibrin (PRF) is a strong but flexible fibrin including a enrich platelet which contain growth factors and cytokines. PRF can be made very simply and requires no artificial additives unlike platelet-rich plasma. While PRF is remodeled and released in the tissue, this induces cell growth, vascularization, collagen synthesis, osteoblast differentiation and an anti-inflammatory reaction. Taking advantage of these functions, PRF can stimulate regeneration of bone and soft tissue in a diverse number of ways during the course of hemostasis, wound coverage, preservation, and reconstruction of alveolar bone. Moreover, the use of PRF to improve bone regeneration has become a recent technique in implantology. In this study, through a literature review of PRF's existing clinical applications, we classified a range of potential PRF oral and maxillofacial surgery applications including preservation of extraction sockets, guided bone graft, sinus lift, dressing and periodontal treatment. This trial gave us chance to confirm the usefulness of PRF. Recently, updated clinical studies results concerning skin and tendon wound healing have become available. These results suggest that the usage of RPF will gradually expand.

• 대한치과보철학회지
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• 제45권1호
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• pp.71-84
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• 2007

Statement of problem: Implant surface characteristics plays an important role in clinical success and many studies have been made for improvement of success by changing surface roughness. Purpose: Appropriate increase of surface roughness increases the activity of osteoblast and enhance contact and retention between bone and implant. Material and method- Machined, SLA and RBM surface implants, which are the most commonly used implants were implanted into the tibia of rabbits and after 1 week, 4 weeks, 8 weeks and 12 weeks there were histologic and histomorphometric analysis and study for bone gradient and change of Ca/P ratio using EDS(Energy Dispersive X-ray Spectroscope). Results: Comparison of bone-implant contact showed no significant difference among each implant. In comparison of bone area rates, SLA showed higher value with significant difference at 1 week and 4 weeks, and SLA and RBM at 8 weeks than Machined implant (p<0.05). In analysis of bone constituents with EDS, titanium was specifically detected in new bones and the rates were constant by surface treatment method or period. In case of Ca/P ratio, according to surface treatment method, each group showed significant difference. Lots of old bone fragments produced during implantation remained on the rough surface of RBM implant surface and each group showed histological finding with active synthesis of collagen fibers until 12 weeks. In transmission electronic microscopic examination of sample slice after elapse of twelve weeks, tens nm of borderline (lamina limitans like dense line)was seen to contact the bone, on the interface between bone and implant. Conclusion: SLA and RBM implant with rough surface shows better histomorphometrical result and the trend of prolonged bone formation and maturation in comparison with Machined implant. In addition, implant with rough surface seems to be helpful in early stage bone formation due to remaining of old bone fragments produced in implantation. From the results above, it is considered to be better to use implant with rough surface in implantation.

• Journal of Periodontal and Implant Science
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• 제31권4호
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• pp.823-832
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• 2001

Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan(poly-N-acetyl glucosaminoglycan), a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the human periodontal ligament fibroblasts(hPDLFs) in vitro, with special focus on their proliferative properties by M'IT assay, the synthesis of type I collagen by reverse transcription-polymerase chain reaction(RT-PCR) and the activity of alkaline phosphatase(ALP). Fibroblast populations were obtained from individuals with a healthy periodontium and cultured with ${\alpha}MEM$ as the control group. The experimental groups were cultured with chitosan in concentration of 0.01,0.1, 1,2mg/ml. The results are as follows; 1. Chitosan-induced proliferative responses of hPDLFs reached a plateau at the concentration of O.lmg/ml(p<0.05). 2. When hPDLFs were stimulated with 0.lmg/ml chitosan, mRNA expression of type I collagen was up-regulated. 3. When hPDLFs were stimulated with 0.lmg/ml chitosan, ALP activity was significantly up-regulated(p<0.05). In summary, chitosan(0.lmg/ml) enhanced the type I collagen synthesis in the early stage, and afterwards, facilitated differentiation into osteogenic cells. The results of this in vitro experiment suggest that chitosan potentiates the differentiation of osteoprogenitor cells and may facilitate the formation of bone.

• Journal of Microbiology and Biotechnology
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• 제31권10호
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• pp.1401-1408
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• 2021

This study examined whether the oral administration of low-molecular-weight collagen peptide (LMCP) containing 3% Gly-Pro-Hyp with >15% tripeptide (Gly-X-Y) content could ameliorate osteoarthritis (OA) progression using a rabbit anterior cruciate ligament transection (ACLT) model of induced OA and chondrocytes isolated from a patient with OA. Oral LMCP administration (100 or 200 mg/kg/day) for 12 weeks ameliorated cartilage damage and reduced the loss of proteoglycan compared to the findings in the ACLT control group, resulting in dose-dependent (p < 0.05) improvements of the OARSI score in hematoxylin & eosin (H&E) and Safranin O staining. In micro-computed tomography analysis, LMCP also significantly (p < 0.05) suppressed the deterioration of the microstructure in tibial subchondral bone during OA progression. The elevation of IL-1β and IL-6 concentrations in synovial fluid following OA induction was dose-dependently (p < 0.05) reduced by LMCP treatment. Furthermore, immunohistochemistry illustrated that LMCP significantly (p < 0.05) upregulated type II collagen and downregulated matrix metalloproteinase-13 in cartilage tissue. Consistent with the in vivo results, LMCP significantly (p < 0.05) increased the mRNA expression of COL2A1 and ACAN in chondrocytes isolated from a patient with OA regardless of the conditions for IL-1β induction. These findings suggest that LMCP has potential as a therapeutic treatment for OA that stimulates cartilage regeneration.

• Biomolecules & Therapeutics
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• 제24권4호
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• pp.395-401
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• 2016

Endochondral bone formation is the process by which mesenchymal cells condense into chondrocytes, which are ultimately responsible for new bone formation. The processes of chondrogenic differentiation and hypertrophy are critical for bone formation and are therefore highly regulated. The present study was designed to investigate the effect of aloe-emodin on chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Aloe-emodin treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. ATDC5 cells were treated with aloe-emodin and stained with alcian blue. Compared with the control cells, the ATDC5 cells showed more intense alcian blue staining. This finding suggested that aloe-emodin induced the synthesis of matrix proteoglycans and increased the activity of alkaline phosphatase. Aloe-emodin also enhanced the expressions of chondrogenic marker genes such as collagen II, collagen X, BSP and RunX2 in a time-dependent manner. Furthermore, examination of the MAPK signaling pathway showed that aloe-emodin increased the activation of extracellular signal-regulated kinase (ERK), but had no effect on p38 and c-jun N-terminal kinase (JNK). Aloe-emodin also enhanced the protein expression of BMP-2 in a time-dependent manner. Thus, these results showed that aloe-emodin exhibited chodromodulating effects via the BMP-2 or ERK signaling pathway. Aloe-emodin may have potential future applications for the treatment of growth disorders.

• 치위생과학회지
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• 제13권2호
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• pp.158-164
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• 2013

임플란트 식립 시 가장 빈번하게 맞게 되는 문제점으로 임플란트 식립 부위에서의 불충분한 골량과 해부학적 구조에 의한 접근성의 문제를 들 수 있다. 일반적으로 성장 인자들은 치유 과정이나 조직 형성에 있어서 가장 기본적인 필수 요소로 인정되고 있다. 이러한 이유로 골 이식 재료의 효과를 증진시키기 위한 성장 인자들이 최근에 주목을 받고 있다. 혈소판 내 granules에는 높은 농도의 다양한 성장 인자들이 포함되어 있다. 특히, platelet-rich fibrin (PRF)는 2세대 혈소판 농축 인자로 항응고제가 들어있지 않은 상태로 얻을 수가 있고, 혈소판과 많은 성장 인자들이 풍부한 섬유소 막을 포함하고 있다. 이번 연구의 목적은 in vitro 상에서 골아 세포에 대한 PRF의 영향을 알아보고자 하였다. 특히 치유와 재생에 연관된 주요 기능으로써 증식과 분화에 대한 영향을 조사하고자 하였다. 이를 위해서, PRF 내에서 방출되는 성장 인자(platelet-derived growth factor subunit B와 transforming growth factor-${\beta}1$)의 농도, 세포의 생존능력, alkaline phosphatase (ALP) activity, type 1 collagen 합성, 골아 세포의 분화 지표로써 ALP와 Runx2의 발현 정도와 골 기질 단백질로써 type 1 collagen의 발현 정도에 대해서 조사하였다. 이 실험을 통하여 PRF는 치유 시 필요한 타당한 기간 동안에 충분히 자가 성장 인자의 방출을 유지하고 있음을 알 수 있었고, 골아 세포의 증식과 분화에 대해서 긍정적인 효과가 있음을 보여 주였다. 제한적인 실험이지만, 골재생을 위한 PRF의 사용은 골 치유와 골 개조에 있어서 증진 효과를 가져다줄 수 있는 촉망되는 방법 중 하나가 될 수 있을 것이다.

• 생명과학회지
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• 제21권12호
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• pp.1752-1760
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• 2011

우리나라에서 자생하고 있는 천년초는 새로운 생리활성 물질을 생산할 수 있는 소재로 각광받고 있으며, mouse calvaria 유래의 MC3T3-E1 세포는 골세포의 세포 활성과 관련된 연구에서 유용하게 이용되어 왔다. 따라서 본 연구에서는 MC3T3-E1 세포를 이용하여 천년초 추출물이 세포 증식에 미치는 영향과 ALP 활성, 조골세포의 골 형성을 위한 필수 인자인 collagen 합성에 대한 영향을 검토하고 세포사의 주요 인자인 ROS 에 미치는 영향에 대해 검토하였다. 각 추출물의 수율은 껍질 열수 추출물이 35.2%로 가장 수율이 높았고, 다음으로 줄기 열수 추출물, 껍질 에탄올 추출물, 씨 열수 추출물, 줄기 에탄올 추출물 순으로 나타났으며, 수율이 가장 낮은 씨 에탄올 추출물은 20.6%였다. 각 추출물의 농도(1, 10 50, $100{\mu}g/ml$)에 따른 조골세포 성장에 미치는 영향을 MTT assay로 분석한 결과, 모든 군에서 대조군과 비교하여 유의적인 증식률을 나타내었으며 특히, $100{\mu}g/ml$ 씨 열수 추출물을 첨가하였을 때, 대조군과 비교하여 가장 높은 120% 정도의 증식률을 나타내었다. 천년초 추출물이 ALP 활성에 미치는 영향을 조사한 결과, 천년초 추출물을 $10\sim50{\mu}g/ml$ 첨가하였을 때, 대조군과 비교하여 유의적으로 증가하였으며 특히, 씨 에탄올 추출물을 $50{\mu}g/ml$ 첨가하였을 때, 130% 이상의 ALP 활성을 증가시켜 조골세포의 분화에 영향을 줄 가능성이 제시 되었다. 천년초 추출물이 조골세포의 collagen 합성에 미치는 실험결과에서는 천년초 씨 열수 추출물과 씨 에탄올 추출물의 $50\sim100{\mu}g/ml$ 농도에서 높은 합성능을 나타내었으며 그 중 씨 열수 추출물 $100{\mu}g/ml$ 농도에서 가장 높은 collagen 합성능을 보였다. 천년초 추출물이 세포내 ROS생성에 미치는 영향을 실험한 결과에서는 모든 천년초 추출물 처리에 의해 농도 의존적으로 형광 강도가 감소하는 경향을 보였으며 특히, 씨 열수 추출물의 $100{\mu}g/ml$ 경우, 대조군에 비해 54% 정도 ROS가 감소되어 천년초씨 추출물이 세포 내에서 높은 항산화력이 있는 것을 확인할 수 있었다. 따라서 천년초 추출물이 조골세포의 증식, ALP 활성, collagen 합성 및 ROS 생성 저해를 촉진하여 골 생성에 영향을 줄 수 있는 것이 확인되었으며, 추출 용매와 관계없이 씨 추출물이 가장 높은 활성을 나타내었다. 천년초 씨 중의 활성 성분 구명을 위해 향후 구체적 기작 연구와 in vivo 연구가 병행된다면, 골다공증 예방과 관련된 기능성 식품의 천연소재 개발이 가능할 것이라 사료된다.

• Journal of Acupuncture Research
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• 제23권2호
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• pp.103-111
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• 2006

Objectives : Deer antler Water Extract(DAE), prepared from the pilose antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe (Nokyong), a traditional immuno-suppressive and immuno-activating Korean herbal-acupuncture, is thought to play an important role in human bone remodeling. Methods : To determine whether DAE can induce the differentiation of resting zone chondrocytes(RC) or not, confluent cell cultures were pretreated for 24, 36, 48, 72, and 120hrs with DAE. At the end of pretreatment, the media were replaced with new media containing $10^{-10}{\sim}10^{-8}M\;1,25-(OH)_2D_3$ and the cells incubated for an additional 24hrs. Results : This second treatment was chosen because prior studies had shown that only the more mature growth zone chondrocytes(GC) respond to this vitamin $D_3$ metabolite. The effect of DAE pretreatment on cell maturation was confirmed by measuring alkaline phosphatase (ALPase)-specific activity. Changes in matrix protein synthesis were examined by measuring collagen synthesis, as well as $^{35}SO_4$ incorporation into proteoglycans. When RC cells were pretreated for 120h with DAE, treatment with $1,25-(OH)_2D_3$ caused a dose-dependent increase in ALPase-specific activity and collagen synthesis, however, the proteoglycan production was not affected. RC cells pretreated with $1,25-(OH)_2D_3$ responded like RC cells that had not received any pretreatment. Conclusion : These results indicate that DAE directly regulates the maturation of RC chondrocytes into GC chondrocytes. Therefore it was indicated that DAE may play a significant role in regulating chondrocyte maturation during endochondral ossification.