• 대한음성언어의학회:학술대회논문집
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• 대한음성언어의학회 2003년도 제19회 학술대회
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• pp.183-183
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• 2003

Background and Objectives : Cartilage reconstruction is one of medical issue in otolaryngology. Tissue engineering is presently being utilized in part of cartilage repair. Sources of cells for tissue engineering are chondrocyte from mature cartilage and bone marrow mesenchymal stem cells that are able to differentiate into chondrocyte. Recent studies have shown that adipose tissue have mesenchymal stem cells which can differentiate into adipogenic, chondrogenic myogenic osteogenic cells and neural cell in vitro. In this study, we have examined chondrogenic potential of the canine adipose tissue-derived mesenchymal stem cell(ATSC). Materials and Methods : We harvested canine adipose tissue from inguinal area. ATSCs were enzymatically released from canine adipose tissue. Under appropriate culture conditions, ATSCs were induced to differentiate into the chondrocyte lineages using micromass culture technique. We used immunostain to type II collagen and toluidine blue stain to confirm chondrogenic differentiation of ATSCs. Results : We could isolate ATSCs from canine adipose tissue. ATSCs expressed CD29 and CD44 which are specific surface markers of mesenchymal stem cell. ATSCs differentiated into micromass that has positive response to immunostain of type II collagen and toluidine blue stain. Conclusion : In vitro, ATSCs differentiated into cells that have characteristic cartilage matrix molecules in the presence of lineage-specific induction factors. Adipose tissue may represent an alternative source to bone marrow-derived MSCs.

• Restorative Dentistry and Endodontics
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• 제37권1호
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• pp.34-40
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• 2012

Objectives: The aim was to confirm the stem cell-like properties of the dental pulp stromal cells and to evaluate the morphologic changes during in vitro chondrogenesis. Materials and Methods: Stromal cells were outgrown from the dental pulp tissue of the premolars. Surface markers were investigated and cell proliferation rate was compared to other mesenchymal stem cells. Multipotency of the pulp cells was confirmed by inducing osteogenesis, adipogenesis and chondrogenesis. The morphologic changes in the chondrogenic pellet during the 21 day of induction were evaluated under light microscope and transmission electron microscope. TUNEL assay was used to evaluate apoptosis within the chondrogenic pellets. Results: Pulp cells were CD90, 105 positive and CD31, 34 negative. They showed similar proliferation rate to other stem cells. Pulp cells differentiated to osteogenic, adipogenic and chondrogenic tissues. During chondrogenesis, 3-dimensional pellet was created with multi-layers, hypertrophic chondrocyte-like cells and cartilage-like extracellular matrix. However, cell morphology became irregular and apoptotic cells were increased after 7 day of chondrogenic induction. Conclusions: Pulp cells indicated mesenchymal stem cell-like characteristics. During the in vitro chondrogenesis, cellular activity was superior during the earlier phase (within 7 day) of differentiation.

• International Journal of Oral Biology
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• 제38권3호
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• pp.121-126
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• 2013

Tumor necrosis factor alpha ($TNF{\alpha}$) is a multifunctional inflammatory cytokine that regulates various cellular and biological processes. Increased levels of $TNF{\alpha}$ have been implicated in a number of human diseases including diabetes and arthritis. Sympathetic nervous system stimulation via the beta2-adrenergic receptor (${\beta}2AR$) in osteoblasts suppresses osteogenic activity. We previously reported that $TNF{\alpha}$ upregulates ${\beta}2AR$ expression in murine osteoblastic cells and that this modulation is associated with $TNF{\alpha}$ inhibition of osteoblast differentiation. In our present study, we explored whether $TNF{\alpha}$ induces ${\beta}2AR$ expression in human osteoblasts and then identified the downstream signaling pathway. Our results indicated that ${\beta}2AR$ expression was increased in Saos-2 and C2C12 cells by $TNF{\alpha}$ treatment, and that this increase was blocked by the inhibition of NF-${\kappa}B$ activation. Chromatin immunoprecipitation and luciferase reporter assay results indicated that NF-${\kappa}B$ directly binds to its cognate elements on the ${\beta}2AR$ promoter and thereby stimulates ${\beta}2AR$ expression. These findings suggest that the activation of $TNF{\alpha}$ signaling in osteoblastic cells leads to an upregulation of ${\beta}2AR$ and also that $TNF{\alpha}$ induces ${\beta}2AR$ expression in an NF-${\kappa}B$-dependent manner.

• 대한치과의사협회지
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• 제53권1호
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• pp.28-35
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• 2015

To overcome shortcoming of autogeneous, allogenic, xenogenic and alloplastic bone grafts, various growth factors related to bone regeneration have been identified and developed. Among them, rhBMP-2 is regarded as the most potent osteoinductive growth factor and it can trigger the differentiation of mesenchymal stem cells to osteogenic cells for accelerated new bone formation And several commercial products of rhBMP-2 are available in Korea. It is applied to maxillary sinus augmentation, guided bone regeneration and preservation of extraction socket. In this review, the development, action mechanism and clinical applications of rhBMP-2 will be described.

• 대한기계학회논문집B
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• 제35권11호
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• pp.1237-1242
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• 2011

인산칼슘 재료는 하이드록시 아파타이트(Hydroxyapatite)와 트리칼슘 포스페이트(Tricalcium-phosphate)를 포함하고 있으며, 인체 골 조직의 무기성분으로 세포 독성이 없고 생체 적합한 성질을 가지고 골 전도성이 있다. 또한 두 재료가 혼합되어 있는 이상 인산칼슘(Biphasic calcium phosphate) 재료는 골 유도성이 있다고 알려져 있다. 이러한 골 조직 재생에 많은 장점을 가지고 있는 인산칼슘 재료는 파우더 타입으로, 3 차원 자유형상의 인공지지체를 제작하는 데 어려움이 있어 고분자 재료에 첨가하여 사용되었다. 본 연구에서는 자유형상 제작 기술을 이용하여 원하는 내/외부 형상을 가지는 3 차원 인산칼슘 인공지지체를 제작하고, 골 조직 재생용 인공지지체로의 사용이 적합한지를 확인하기 위해 MC3T3-E1 를 이용한 세포 증식, 골 조직 분화 실험을 수행하였다.

• The Journal of Advanced Prosthodontics
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• 제2권1호
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• pp.7-13
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• 2010

Although most researchers agree that platelet-rich plasma (PRP) is a good source of autogenous growth factors, its effect on bone regeneration is still controversial. The purpose of this study was to evaluate whether increasing angiogenic factors in the human PRP to enhance new bone formation through rapid angiogenesis. MATERIAL AND METHODS. In vitro, the human platelets were activated with application of shear stress, $20\;{\mu}g/ml$ collagen, 2 mM $CaCl_2$ and 10U thrombin/$1\;{\times}\;10^9$ platelets. Level of vascular endothelial growth factor (VEGF) and platelet microparticle (PMP) in the activated platelets were checked. In the animal study, human angiogenic factors-enriched PRP was tested in 28 athymic rat's cranial critical bone defects with $\beta$-TCP. Angiogenesis and osteogenesis were evaluated by laser Doppler perfusion imaging, histology, dual energy X-ray densinometry, and micro-computed tomography. RESULTS. In vitro, this human angiogenic factors-enriched PRP resulted in better cellular proliferation and osteogenic differentiation. In vivo, increasing angiogenic potential of the PRP showed significantly higher blood perfusion around the defect and enhanced new bone formation around acellular bone graft material. CONCLUSION. Angiogenic factor-enriched PRP leads to faster and more extensive new bone formation in the critical size bone defect. The results implicate that rapid angiogenesis in the initial healing period by PRP could be supposed as a way to overcome short term effect of the rapid angiogenesis.

• 대한의용생체공학회:의공학회지
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• 제43권1호
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• pp.61-70
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• 2022

Natural and synthetic forms of calcium phosphate cement (CPC) have been widely used in bone repair and augmentation. The major challenge of injectable CPC is to deliver the cells without cell death in order to regenerate new bone. The study objective was to investigate for the potential of stem cell-laden gelatin fibers containing injectable, nanocrystalline CPC to function as a delivery system. Gelatin noddle fiber method was developed to delivered cells into nCPC. Experimental groups were prepared by mixing cells with nCPC, mixing cell-laden gelatin fibers with nCPC and mixing cell-laden gelatin fibers containing BMP-2 with nCPC. Media diffusion test was conducted after dissolving the gelatin fibers. SEM examined the generated channels and delivered cell morphology. Fibers mixed with nCPC showed physical setting and hardening within 20 min after injection and showed good shape maintenances. The gelatin fibers mixed nCPC group had several vacant channels generated from the dissolved gelatin. Particularly, proliferation and attachment of the cells were observed inside of the channels. While live cells were not observed in the cell mixed nCPC group, cells delivered with the gelatin fibers into the nCPC showed good viability and increased DNA content with culture. Cell-laden gelatin fiber was a novel method for cell delivery into nCPC without cell damages. Results also indicated the osteogenic differentiation of gelatin fiber delivered cells. We suggest that the cell-laden gelatin fibers mixed with nCPC can be used as an injectable cell delivery vehicle and the addition of BMP-2 to enhances osteogenesis.

• Journal of Periodontal and Implant Science
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• 제52권2호
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• pp.155-169
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• 2022

Purpose: The aim of this study was to determine the effect of insulin growth factor binding protein-3 (IGFBP-3) on the inhibition of glucose oxidative stress and promotion of bone formation near the implant site in a rat model of methylglyoxal (MGO)-induced bone loss. Methods: An in vitro study was performed in MC3T3 E1 cells treated with chitosan gold nanoparticles (Ch-GNPs) conjugated with IGFBP-3 cDNA followed by MGO. An in vivo study was conducted in a rat model induced by MGO administration after the insertion of a dental implant coated with IGFBP-3. Results: MGO treatment downregulated molecules involved in osteogenic differentiation and bone formation in MC3T3 E1 cells and influenced the bone mineral density and bone volume of the femur and alveolar bone. In contrast, IGFBP-3 inhibited oxidative stress and inflammation and enhanced osteogenesis in MGO-treated MC3T3 E1 cells. In addition, IGFBP-3 promoted bone formation by reducing inflammatory proteins in MGO-administered rats. The application of Ch-GNPs conjugated with IGFBP-3 as a coating of titanium implants enhanced osteogenesis and the osseointegration of dental implants. Conclusions: This study demonstrated that IGFBP-3 could be applied as a therapeutic component in dental implants to promote the osseointegration of dental implants in patients with diabetes, which affects MGO levels.

• Journal of Periodontal and Implant Science
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• 제53권4호
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• pp.259-268
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• 2023

Purpose: Hyaluronic acid (HA) affects angiogenesis and promotes the migration and differentiation of mesenchymal cells, thereby activating the osteogenic ability of osteoblasts. Although studies on the action of HA during bone regeneration are being actively conducted, the optimal dose of HA required for bone regeneration remains unclear. Therefore, the purpose of this study was to elucidate the most effective HA dose for bone formation using a rat critical-size defect model. Methods: Thirty rats were randomly divided into 5 groups, with 6 rats in each group. An absorbable collagen sponge soaked with HA or saline was used to fill an 8-mm defect, which was then covered with a collagen membrane. Different treatments were performed for each group as follows: (1) saline control, (2) 1 mg/mL HA, (3) 25 mg/mL HA, (4) 50 mg/mL HA, or (5) 75 mg/mL HA. After a healing period of 4 weeks, micro-computed tomography and histological analysis were performed. The obtained values were analyzed using analysis of variance and the Tukey test (P<0.05). Results: At week 4, the 75 mg/mL HA group had the highest bone volume/total volume ratio, new bone, and bone fill among the 5 groups, and these values were significantly different from those observed in the control group (P<0.01) and 1 mg/mL HA group (P<0.001). More active bone formation was observed in the higher-dose HA groups (25 mg/mL, 50 mg/mL, and 75 mg/mL HA), which included a large amount of woven bone. Conclusions: The 75 mg/mL HA group showed better bone formation than the other groups (1, 25, and 50 mg/mL HA and control).

• Journal of Microbiology and Biotechnology
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• 제34권2호
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• pp.415-424
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• 2024

This study reveals that low-molecular-weight collagen peptide (LMWCP) can stimulate the differentiation and the mineralization of MC3T3-E1 cells in vitro and attenuate the bone remodeling process in ovariectomized (OVX) Sprague-Dawley rats in vivo. Moreover, the assessed LMWCP increased the activity of alkaline phosphatase (ALP), synthesis of collagen, and mineralization in MC3T3-E1 cells. Additionally, mRNA levels of bone metabolism-related factors such as the collagen type I alpha 1 chain, osteocalcin (OCN), osterix, bone sialoprotein, and the Runt family-associated transcription factor 2 were increased in cells treated with 1,000 ㎍/ml of LMWCP. Furthermore, we demonstrated that critical bone morphometric parameters exhibited significant differences between the LMWCP (400 mg/kg)-receiving and vehicle-treated rat groups. Moreover, the expression of type I collagen and the activity of ALP were found to be higher in both the femur and lumbar vertebrae of OVX rats treated with LMWCP. Finally, the administration of LMWCP managed to alleviate osteogenic parameters such as the ALP activity and the levels of the bone alkaline phosphatase, the OCN, and the procollagen type 1 N-terminal propeptide in OVX rats. Thus, our findings suggest that LMWCP is a promising candidate for the development of food-based prevention strategies against osteoporosis.