• International Journal of Industrial Entomology and Biomaterials
• /
• 제27권2호
• /
• pp.303-311
• /
• 2013

In this study, we compared the efficiency of osteoblast differentiation media (ODM) containing three distinct reagent combinations in osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) in monolayer culture. In addition, we analyzed growth and differentiation of hBMSCs on silk scaffolds and examined the bone-forming activity of a nanofibrous silk scaffold in a tibia diaphysis defect model of a rat hind limb with intramedullary nailing. Although all three ODM increased alkaline phosphatase activity to a comparable extent, the ODM containing bone morphogenetic protein-2 (BMP-2) was found to be significantly less effective in promoting mineral deposition than the others. Growth of hBMSCs on sponge-form silk scaffolds was faster than on nanofibrous ones, while osteoblastic differentiation was apparent in the cells grown on either type of scaffold. By contrast, bone formation was observed only at the edge of the nanofibrous scaffold implanted in the tibia diaphysis defect, suggesting that use of the silk scaffold alone is not sufficient for the reconstitution of the long bone defect. Since silk scaffolds can support cell growth and differentiation in vitro, loading MSCs on scaffolds might be necessary to improve the bone-forming activity of the scaffold in the long bone defect model.

• Journal of Veterinary Science
• /
• 제19권6호
• /
• pp.735-743
• /
• 2018

We investigated the effect of transforming growth factor beta 1 ($TGF-{\beta}1$) on equine hyaluronan synthase 2 (HAS2) gene expression and hyaluronan (HA) synthesis in culture models of articular chondrocytes. Equine chondrocytes were treated with $TGF-{\beta}1$ at different concentrations and times in monolayer cultures. In three-dimensional cultures, chondrocyte-seeded gelatin scaffolds were cultured in chondrogenic media containing 10 ng/mL of $TGF-{\beta}1$. The amounts of HA in conditioned media and in scaffolds were determined by enzyme-linked immunosorbent assays. HAS2 mRNA expression was analyzed by semi-quantitative reverse transcription polymerase chain reaction. The uronic acid content and DNA content of the scaffolds were measured by using colorimetric and Hoechst 33258 assays, respectively. Cell proliferation was evaluated by using the alamarBlue assay. Scanning electron microscopy (SEM), histology, and immunohistochemistry were used for microscopic analysis of the samples. The upregulation of HAS2 mRNA levels by $TGF-{\beta}1$ stimulation was dose and time dependent. $TGF-{\beta}1$ was shown to enhance HA and uronic acid content in the scaffolds. Cell proliferation and DNA content were significantly lower in $TGF-{\beta}1$ treatments. SEM and histological results revealed the formation of a cartilaginous-like extracellular matrix in the $TGF-{\beta}1$-treated scaffolds. Together, our results suggest that $TGF-{\beta}1$ has a stimulatory effect on equine chondrocytes, enhancing HA synthesis and promoting cartilage matrix generation.

• 생체재료학회지
• /
• 제22권4호
• /
• pp.279-289
• /
• 2018

Background: Cell behavior is the key to tissue regeneration. Given the fact that most of the cells used in tissue engineering are anchorage-dependent, their behavior including adhesion, growth, migration, matrix synthesis, and differentiation is related to the design of the scaffolds. Thus, characterization of the scaffolds is highly required. Micro-computed tomography (micro-CT) provides a powerful platform to analyze, visualize, and explore any portion of interest in the scaffold in a 3D fashion without cutting or destroying it with the benefit of almost no sample preparation need. Main body: This review highlights the relationship between the scaffold microstructure and cell behavior, and provides the basics of the micro-CT method. In this work, we also analyzed the original papers that were published in 2016 through a systematic search to address the need for specific improvements in the methods section of the papers including the amount of provided information from the obtained results. Conclusion: Micro-CT offers a unique microstructural analysis of biomaterials, notwithstanding the associated challenges and limitations. Future studies that will include micro-CT characterization of scaffolds should report the important details of the method, and the derived quantitative and qualitative information can be maximized.

• Macromolecular Research
• /
• 제15권5호
• /
• pp.424-429
• /
• 2007

A procedure for the surface hydrolysis of an electrospun poly(${\epsilon}$-caprolactone) (PCL) fibrous scaffold was developed to enhance the adhesion and proliferation of osteoblasts. The surface hydrolysis of fibrous scaffolds was performed using NaOH treatment for the formation of carboxyl groups on the fiber surfaces. The hydrolysis process did not induce deformation of the fibers, and the fibers retained their diameter. The cell seeding density on the NaOH-treated PCL fibrous scaffolds was more pronounced than on the non-treated PCL fibers used as a control. The alkaline phosphatase activity, osteocalcin and a mineralization assay strongly supported that the surface-hydrolyzed PCL fibrous scaffolds provided more favorable environments for the proliferation and functions of osteoblasts compared to the non-treated PCL fibrous scaffolds use as a control.

• 한국정밀공학회:학술대회논문집
• /
• 한국정밀공학회 2005년도 추계학술대회 논문집
• /
• pp.231-235
• /
• 2005

Most tissue engineering strategies for creating functional replacement tissues or organs rely on the application of temporary three-dimensional scaffolds to guide the proliferation and spread of seeded cells in vitro and in vivo. Scaffolds should be satisfied following requirements; macrostructure to promote cell proliferation, pore interconnectivity, pore size ranging from 200 to $400{\mu}m$, surface chemistry and mechanical properties. Rapid prototyping techniques have often been used as an useful process that fabricates scaffolds with complex structures. In this study, a new process to fabricate a three-dimensional scaffolds using bio-compatible polymer has been developed. It employs a highly accurate three-dimensional positioning system with pressure-controlled syringe to deposit biopolymer structures. The pressure-activated microsyringe is equipped with fine-bore nozzles of various inner-diameters. In order to examine relationships between line width and process parameters such as nozzle height, applied pressure, and speed of needle, experiments were carried out. Based on the experimental results, three-dimensional scaffold was fabricated using the apparatus. It shows the validity of the proposed process.

• 폴리머
• /
• 제29권2호
• /
• pp.198-203
• /
• 2005

용융 압출 발포에 의하여 폴리락틱산(PLLA) 지지체를 제조하고 발포 조건이 지지체의 구조와 기계적 특성에 미치는 영향을 살펴보고 이를 염 추출법에 의하여 제조된 지지체와 비교하였다. 발포제를 함유한 PLLA 용융체가 압출기 및 다이에서 체류하는 시간이 발포제의 무게를 최대로 감소시키는 시간과 일치하여야 최적의 PLLA 지지체의 구조를 얻을 수 있음을 확인하였다. PLLA지지체의 구조를 유지하기 위해서는 발포제의 함량을 $10\;wt\%$ 이하로 조절해야 하며 PLLA 지지체의 다공도는 PLLA의 압출기에서의 체류시간에 가장 큰 영향을 받음을 알 수 있었다. 용융 압출 발포에 의하여 얻어진 지지체는 일반적으로 염 추출법에 의하여 제조된 지지체보다 다소 낮은 다공도를 갖지만 발포 조건의 조절에 의하여 적절한 다공 크기와 다공의 연결성을 가지며 동시에 우수한 기계적 특성을 가져 경조직 재생(hard tissue regeneration)용으로 사용이 가능할 것으로 사료된다.

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

• Macromolecular Research
• /
• 제10권3호
• /
• pp.158-167
• /
• 2002

In order to endow with new bioactive functionality from small intestine submucosa (SIS) powder as natural source to poly (L-lactide) (PLA) and poly (lactide-co-glycolide) (PLGA) synthetic biodegradable polymer, porous SIS/PLA and SIS/PLGA as natural/synthetic composite scaffolds were prepared by means of the solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. A uniform distribution of good interconnected pores from the surface to core region was observed the pore size of 40~500 ${\mu}{\textrm}{m}$ independent with SIS amount using the solvent casting/salt leaching method. Porosities, specific pore areas as well as pore size distribution also were almost same. After the fabrication of SIS/PLA hybrid scaffolds, the wetting properties was greatly enhanced resulting in more uniform cell seeding and distribution. Five groups as PGA non-woven mesh without glutaraldehyde (GA) treatment, PLA scaffold without or with GA treatment, and SIS/PLA (Code No.3 ; 1 : 12 of salt content, (0.4 : 1 of SIS content, and 144 ${\mu}{\textrm}{m}$ of median pore size) without or with GA treatment were implanted into the back of nude mouse to observe the effect of SIS on the induction of cells proliferation by hematoxylin and eosin, and von Kossa staining for 8 weeks. It was observed that the effect of SIS/PLA scaffolds with GA treatment on bone induction are stronger than PLA scaffolds, that is to say, in the order of PLA/SIS scaffolds with GA treatment > PLA/SIS scaffolds without GA treatment > PGA nonwoven > PLA scaffolds only with GA treatment = PLA scaffolds only without GA treatment for the osteoinduction activity. The possible explanations are (1) many kinds of secreted, circulating, and extracellular matrix-bound growth factors from SIS to significantly affect critical processes of tissue development and differentiation, (2) the exposure of SIS to GA resulted in significantly calcification, and (3) peri-implant fibrosis due to covalent bonding between collagen molecule by crosslinking reaction. In conclusion, it seems that SIS plays an important role for bone induction in SIS/PLA scaffolds for the application of tissue engineering area.

• 폴리머
• /
• 제34권5호
• /
• pp.398-404
• /
• 2010

본 연구팀은 DBP를 함침시킨 물성이 서로 다른 스폰지와 PLGA 지지체를 제작한 후 세포 부착, 증식 및 형태 유지를 알아보기 위한 실험을 수행하였다. WST 분석법과 SEM 관찰을 통하여 스폰지에 비해서 PLGA 지지체에서의 세포의 증식이 활발한 것을 확인하였고, RT-PCR을 통해 디스크세포에서 특이적으로 발현하는 제 2형 콜라겐과 어그리칸의 발현을 확인하였다. WST 결과, 세포 증식률은 DBP/PLGA 지지체가 DBP를 함침시킨 스폰지보다 세포 증식률이 높음을 확인하였다. 본 연구팀은 스폰지보다 PLGA 지지체가 인간디스크의 표현형 유지 및 증식에 있어서 긍정적인 영향을 미치는 것을 확인하였다.

• 폴리머
• /
• 제36권4호
• /
• pp.401-406
• /
• 2012

KUSA-A1 골조세포로부터 골재생을 유도하기 위하여, 스폰지형(CSS) 및 부직포형(CSNW)의 키토산 지지체를 적용하였다. CSNW의 표면적 및 공극 크기는 CSS에 비해 상대적으로 큰 값을 나타낸 반면, 공극 부피는 CSNW의 경우가 CSS에 비해 작은 값을 보였다. 세포고정 시험 결과는 CSNW의 경우가 더 적합한 결과를 나타내었으며, 이는 지지체의 넓은 표면적에 기인한 것으로 판단되었다. In vivo 실험을 위하여 세포를 각각의 지지체에 투여 후 일주일간 배양하였으며, BALB/C 무모생쥐의 피하조직에 이식하였다. 이식된 지지체는 각각 수술후 1, 4, 6 및 8주에 채취되어 면역학적 염색을 실시하였다. CSS는 수술후 4주에서 6주 사이에 붕괴되기는 하였으나, 조직의 안정성은 CSNW에 비해 우수한 것으로 관찰되었다. 골조직의 생성은 CSNW와 CSS에 대해 각각 4주 및 8주에서 이루어짐을 확인하였다.