• 대한기계학회:학술대회논문집
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• 대한기계학회 2007년도 춘계학술대회A
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• pp.1265-1270
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• 2007

Conventional methods for fabricating three-dimensional (3-D) scaffolds have substantial limitations. In this paper, we present 3-D scaffolds that can be made repeatedly with the same dimensions using a microstereolithography system. This system allows the fabrication of a pre-designed internal structure, such as pore size and porosity, by stacking photopolymerized materials. The scaffolds must be manufactured in a material that is biocompatible and biodegradable. In this regard, we synthesized liquid photocurable biodegradable TMC/TMP, followed by acrylation at terminal ends. And also, solidification properties of TMC/TMP polymer are to be obtained through experiments. Cell adhesion to scaffolds significantly affects tissue regeneration. As a typical example, we seeded chondrocytes on two types of 3-D scaffold and compared the adhesion results. Based on these results, the scaffold geometry is one of the most important factors in chondrocyte adhesion. These 3-D scaffolds could be key factors for studying cell behavior in complex environments and eventually lead to the optimum design of scaffolds for the regeneration of various tissues, such as cartilage and bone.

• 한국표면공학회:학술대회논문집
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• 한국표면공학회 2016년도 추계학술대회 논문집
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• pp.199-199
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• 2016

Commercially pure titanium (cp-Ti) and Ti alloys (typically Ti-6Al-4V) display excellent corrosion resistance and biocompatibility. Although the chemical composition and topography are considered important, the mechanical properties of the material and the loading conditions in the host have, conventionally. Ti and its alloys are not bioactive. Therefore, they do not chemically bond to the bone, whereas they physically bond with bone tissue. The electrochemical deposition process provides an effective surface for biocompatibility because large surface area can be served to cell proliferation. Electrochemical deposition method is an attractive technique for the deposition of hydroxyapatite (HAp). However, the adhesions of these coatings to the Ti surface needs to be improved for clinical used. Plasma electrolyte oxidation (PEO) enables control in the chemical com position, porous structure, and thickness of the $TiO_2$ layer on Ti surface. In addition, previous studies h ave concluded that the presence of $Ca^{+2}$ and ${PO_4}^{3-}$ ion coating on porous $TiO_2$ surface induced adhesion strength between HAp and Ti surface during electrochemical deposition. Silicon (Si) in particular has been found to be essential for normal bone and cartilage growth and development. Zinc (Zn) plays very important roles in bone formation and immune system regulation, and is also the most abundant trace element in bone. The objective of this work was to study electrochemical characteristcs of Zn and Si coating on Ti-6Al-4V by PEO treatment. The coating process involves two steps: 1) formation of porous $TiO_2$ on Ti-6Al-4V at high potential. A pulsed DC power supply was employed. 2) Electrochemical tests were carried out using potentiodynamic and AC impedance methoeds. The morphology, the chemical composition, and the micro-structure an alysis of the sample were examined using FE-SEM, EDS, and XRD. The enhancements of the HAp forming ability arise from $Si/Zn-TiO_2$ surface, which has formed the reduction of the Si/Zn ions. The promising results successfully demonstrate the immense potential of $Si/Zn-TiO_2$ coatings in dental and biomaterials applications.

• 대한의용생체공학회:학술대회논문집
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• 대한의용생체공학회 1998년도 추계학술대회
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• pp.169-170
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• 1998

In this study, regional variation in vertebral bone density due to osteoporosis were investigated using a method that employs images from QCT. QCT images(1mm thick slices) of the first lumbar vertebra from a normal person (23/M, BMD=139.8mg/ml) and from an osteoporotic patient (54/F, BMD=82.0mg/ml) were obtained. Uniform settings (140kVp, 204mA) were used and images of 300 Hounsfield Unit or greater were selectied to filler out soft tissue interference. To assess the regional variation of the area fraction the vertebral body was divided into 3 layers and each layer contained 9 regions. Area faction was calculated based on image analysis data. Our results showed that the area fraction at the middle of the vertebra was quite lower than the endplate and peripheral regions, but the area fraction values from the osteoporotic patient were uniform throughout the entire height of the vertebral body, which indicates the significant drop of BMD had occurred near both end-plates due to the osteoporosis, especially at the peripheral regions. Our results suggest the susceptability of the vertebrae to compression fracture types in osteoporotic spine.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권6호
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• pp.511-519
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• 2006

Autogenous bone grafts have been considered the gold standard for maxillofacial bony defects. However, this procedure could entail a complicated surgical procedure as well as potential donor site morbidity. Possibly the best solution for bone-defect regeneration is a tissue engineering approach, i.e. the use of a combination of a suitable scaffold with osteogenic cells. A major source of osteogenic cells is the bone marrow. Bone marrow-derived mesenchymal stem cells are multipotent and have the ability to differentiate into osteoblastic, chondrocytic, and adipocytic lineage cells. However, the isolation of cells from bone marrow has someproblems when used in clinical setting. Bone marrow aspiration is sometimes potentially more invasive and painful procedure and carries of a risk of morbidity and infection. A minimally invasive, easily accessible alternative would be cells derived from periosteum. The periosteum also contains multipotent cells that have the potential to differentiate into osteoblasts and chondrocytes. In the present study, we evaluated the osteogenic activity and mineralization of cultured human periosteal-derived cells. Periosteal explants were harvested from mandibule during surgical extraction of lower impacted third molar. The periosteal cells were cultured in the osteogenic inductive medium consisting of DMEM supplemented with 10% fetal calf serum, 50g/ml L-ascorbic acid 2-phosphate, 10 nmol dexamethasone and 10 mM -glycerophosphate for 42 days. Periosteal-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 14 of culture period, then decreased in intensity during the culture period. ALP mRNA expression increased up to day 14 with a decrease thereafter. Osteocalcin mRNA expression appeared at day 7 in culture, after that its expression continuously increased in a time-dependent manner up to the entire duration of culture. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. In conclusion, our study showed that cultured human periosteal-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix. As the periosteal-derived cells, easily harvested from intraoral procedure such as surgical extraction of impacted third molar, has the excellent potential of osteogenic capacity, tissue-engineered bone using periosteal-derived cells could be the best choice in reconstruction of maxillofacial bony defects.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권6호
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• pp.495-505
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• 2006

In this study we evaluate the effects of bFGF-BB and PDGF on in vitro proliferation, differentiation and mineralization of mesenchymal stem cells (MSCs) from rat. MSCs were prepared from the bone marrow of 6 or 7-week-old male rats with a technique previously described by Maniatopoulos et al. in 1988. Lineage differentiation to osteogenesis, chondrogenesis and adipogenesis were performed. At first, we characterized the cultured cell on passage 1, 3, 5, 7 with immunocytochemical staining using CD29, 44, 34, 45, ${\alpha}$-SMA and type I collagen. And to study the effects of bFGF and PDGF-BB on proliferation, differentiation and mineralization, we seeded the expanded cell at a density of 6 $6{\times}10^3\;cells/cm^2$ to 100-mm dish for evaluation of cell proliferation and MTT assay was carried out on day 2, 4, 7, 9. We also resuspended the cells with same density $(6{\times}10^3\;cells/cm^2)$ to 24 well plates for subculture. On the following day, the attached cells were exposed to 2.5ng/ml bFGF and/or 25ng/ml PDGF-BB daily during 5 days. The osteocalcin (OC) level was assessed and mineral contents were evaluated with alizarin red S staining on subculture day 2, 7, 14, 21. We identified the mesenchymal stem cell from the bone marrow derived cells of rat through their successful multi-differentiation and stable display of its phenotype. And bFGF and PDGF-BB showed the effect that inhibited osteoblastic differentiation and mineralization mildly in above concentration at in vitro culture. This study was supported by grant 04-2004-0120 from the Seoul National University Hospital Research Fund.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권4호
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• pp.412-418
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• 2008

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 efficient staining pattern of ALP and Alizarin red. The feasibility 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.

• 한국세라믹학회지
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• 제45권10호
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• pp.618-624
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• 2008

Various calcium phosphate bioceramics are distinguished by their excellent biocompatibility and osteoconductivity. Especially, the exceptional biodegradability of $\beta$-TCP makes it a bone graft substitute of choice in many clinical applications. The activation of osteoclasts, differentiated from macrophage precursor cells, trigger a cell-mediated resorption mechanism that renders $\beta$-TCP biodegradable. Based on this evidence, we studied the biodegradation process of granular-type $\beta$-TCP bone graft substitute through in vitro and in vivo studies. Raw 264.7 cells treated with RANKL and M-CSF differentiated into osteoclasts with macrophage-like properties, as observed with TRAP stain. These osteoclasts were cultured with $\beta$-TCP nano powders synthesized by microwave-assisted process. We confirmed the phagocytosis of osteoclasts by observing $\beta$-TCP particles in their phagosomes via electron microscopy. No damage to the osteoclasts during phagocytosis was observed, nor did the $\beta$-TCP powders show any sign of cytotoxicity. We also observed the histological changes in subcutaneous tissues of rats implanted with granule-type $\beta$-TCP synthesized by fibrous monolithic process. The $\beta$-TCP bone graft substitute was well surrounded with fibrous tissue, and 4 months after implantation, 60% of its mass had been biodegraded. Also, histological findings via H&E stain showed a higher level of infiltration of lymphocytes as well as macrophages around the granule-type $\beta$-TCP. From the results, we have concluded that macrophages play an important role in the biodegradation process of $\beta$-TCP bone graft substitutes.

• Asian-Australasian Journal of Animal Sciences
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• 제32권6호
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• pp.896-903
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• 2019

Objective: Qianbei-Pockmarked goats are affected by a disorder locally referred to as 'Ruanguzheng Disorder', which is characterized by emaciation, lameness, muscular relaxation, stiffness of the extremities, and abnormal curvatures of the long bones. Our objective was to determine the relationship between the disorder and phosphorus deficiency. Methods: Tissue samples were collected from affected and healthy animals, while soil and herbage samples were collected from affected and healthy pastures. Biochemical parameters were determined using an automatic biochemical analyzer (OLYMPUS AU 640, Olympus Optical Co., Tokyo, Japan). Mineral contents in soil, forage, and tissue were determined using a Perkin-Elmer AAS5000 atomic absorption spectrophotometer (Perkin-Elmer, Norwalk, CT, USA). Results: The results showed that phosphorus contents in herbages from affected pastures were markedly lower than those from healthy areas (p<0.01), and the ratio of calcium to phosphorus in the affected herbages was 12.93:1. The phosphorus contents of wool, blood, tooth, and bone from affected animals were also markedly lower than those from healthy animals (p<0.01). Serum phosphorus values in affected animals were much lower than those in healthy animals, while serum alkaline phosphatase values from affected animals were markedly higher than those from healthy animals (p<0.01). Inorganic phosphorus values from affected animals were approximately half of that in the control group. Supplementation of disodium hydrogen phosphate prevented and cured the disorder. Conclusion: This study demonstrates that Ruanguzheng disorder in Qianbei-Pockmarked goats is primarily caused by phosphorus deficiencies in herbage due to fenced pastures and natural habitat fragmentation.

• 대한기계학회논문집A
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• 제38권4호
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• pp.371-377
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• 2014

조직 공학에 있어 인공지지체는 손상된 조직 및 기관의 기능을 재생하기 위한 거푸집으로 제공되며 3 차원 구조물이다. 인공지지체의 재료 중에서 폴리카프로락톤(Polycaprolactone, PCL)과 삼인산칼슘(${\beta}$-tricalcium phosphate, ${\beta}$-TCP)은 생분해성과 생체적합성을 가지고 있다. 본 연구에서는 다축 인공지지체 제작 시스템을 이용하여 3 차원 PCL, blended PCL(60 wt %)/${\beta}$-TCP(40 wt %), 그리고 ${\beta}$-TCP 인공지지체를 제작하였다. 제작된 인공지지체는 주사전자현미경 분석을 통해 $600{\pm}20{\mu}m$의 공극 크기로 잘 제작되었다. 기계적 특성 평가를 통해 3 차원 PCL, blended PCL(60 wt %)/${\beta}$-TCP(40 wt %), 그리고 ${\beta}$-TCP 인공지지체의 효과는 분석되었다. 게다가 Saos-2 세포를 이용한 in vitro 연구를 수행하여 세포 증착 및 증식과 같은 세포 거동에 의한 3 차원 인공지지체의 효과를 확인하였다. 요컨대 3D blended PCL(60 wt %)/${\beta}$-TCP(40 wt %) 인공지지체가 압축 강도와 생체적합성 그리고 골전도성에 있어서 인체의 해면골에 더욱 적합하였다. 따라서 3D 인공지지체의 제작에 있어 PCL과 ${\beta}$-TCP를 혼합하는 것은 효과적인 골 재생을 위해 촉망되는 전략이 될 것이다.

• 폴리머
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• 제36권6호
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• pp.789-794
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• 2012

생체적합성 및 생분해성의 장점을 지닌 PLGA는 우수한 기계적 성질과 분해속도를 조절할 수 있는 장점을 가지고 있지만 소수성으로 인해 세포의 초기부착률이 낮고, 지지체 내부에 영양액의 공급이 원활하지 않다. 본 연구에서는 이러한 단점을 보완하고자 탈미네랄화된 골분(DBP, demineralized bone particle) 젤을 PLGA 지지체에 함침시켰다. 다양한 형태의 지지체 압축강도를 측정한 결과 PLGA/DBP 지지체가 연골과 비슷한 강도를 가지고 있었으며, 이를 바탕으로 DBP젤을 함침시킨 PLGA/DBP젤 지지체를 제작하였다. 연골세포를 파종하여 in vivo 실험을 진행하였고 조직화 정도를 확인하기 위해 H&E, Safranin-O, Alcian blue, Collagen type I 및 Collagen type II 염색을 실시하였다. 그 결과 DBP를 함침시킨 PLGA 지지체에서 GAG, Collagen type I 및 Collagen type II의 높은 발현과 조직화를 보여 연골조직으로 대체할 수 있는 가능성을 확인하였다.