• 대한두개안면성형외과학회지
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• 제19권3호
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• pp.181-189
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• 2018

Background: Autogenous bone grafts have several limitations including donor-site problems and insufficient bone volume. To address these limitations, research on bone regeneration is being conducted actively. In this study, we investigate the effects of a three-dimensionally (3D) printed polycaprolactone (PCL)/tricalcium phosphate (TCP) scaffold on the osteogenic differentiation potential of adipose tissue-derived stem cells (ADSCs) and bone marrow-derived stem cells (BMSCs). Methods: We investigated the extent of osteogenic differentiation on the first and tenth day and fourth week after cell culture. Cytotoxicity of the 3D printed $PCL/{\beta}-TCP$ scaffold was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, prior to osteogenic differentiation analysis. ADSCs and BMSCs were divided into three groups: C, only cultured cells; M, cells cultured in the 3D printed $PCL/{\beta}-TCP$ scaffold; D, cells cultured in the 3D printed $PCL/{\beta}-TCP$ scaffold with a bone differentiation medium. Alkaline phosphatase (ALP) activity assay, von Kossa staining, reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting were performed for comparative analysis. Results: ALP assay and von Kossa staining revealed that group M had higher levels of osteogenic differentiation compared to group C. RT-PCR showed that gene expression was higher in group M than in group C, indicating that, compared to group C, osteogenic differentiation was more extensive in group M. Expression levels of proteins involved in ossification were higher in group M, as per the Western blotting results. Conclusion: Osteogenic differentiation was increased in mesenchymal stromal cells (MSCs) cultured in the 3D printed PCL/TCP scaffold compared to the control group. Osteogenic differentiation activity of MSCs cultured in the 3D printed PCL/TCP scaffold was lower than that of cells cultured on the scaffold in bone differentiation medium. Collectively, these results indicate that the 3D printed PCL/TCP scaffold promoted osteogenic differentiation of MSCs and may be widely used for bone tissue engineering.

• Archives of Plastic Surgery
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• 제44권5호
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• pp.370-377
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• 2017

Background Composite grafts are frequently used for facial reconstruction. However, the unpredictability of the results and difficulties with large defects are disadvantages. Adipose-derived stem cells (ADSCs) express several cytokines, and increase the survival of random flaps and fat grafts owing to their angiogenic potential. Methods This study investigated composite graft survival after ADSC injection. Circular chondrocutaneous composite tissues, 2 cm in diameter, from 15 New Zealand white rabbits were used. Thirty ears were randomly divided into 3 groups. In the experimental groups (1 and 2), ADSCs were subcutaneously injected 7 days and immediately before the operation, respectively. Similarly, phosphate-buffered saline was injected in the control group just before surgery in the same manner as in group 2. In all groups, chondrocutaneous composite tissue was elevated, rotated 90 degrees, and repaired in its original position. Skin flow was assessed using laser Doppler 1, 3, 6, 9, and 12 days after surgery. At 1 and 12 days after surgery, the viable area was assessed using digital photography; the rabbits were euthanized, and immunohistochemical staining for CD31 was performed to assess neovascularization. Results The survival of composite grafts increased significantly with the injection of ADSCs (P<0.05). ADSC injection significantly improved neovascularization based on anti-CD31 immunohistochemical analysis and vascular endothelial growth factor expression (P<0.05) in both group 1 and group 2 compared to the control group. No statistically significant differences in graft survival, anti-CD31 neovascularization, or microcirculation were found between groups 1 and 2. Conclusions Treatment with ADSCs improved the composite graft survival, as confirmed by the survival area and histological evaluation. The differences according to the injection timing were not significant.

• International Journal of Stem Cells
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• 제15권3호
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• pp.291-300
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• 2022

Background and Objectives: Many preclinical studies have been conducted using animal disease models to determine the effectiveness of human mesenchymal stem cells (hMSCs) for treating immune and inflammatory diseases based on the belief that hMSCs are not immunogenic across species. However, several researchers have suggested xenogeneic immune responses to hMSCs in animals, still without detailed features. This study aimed to investigate a xenogeneic humoral immune response to hMSCs in mice in detail. Methods and Results: Balb/c mice were intraperitoneally injected with adipose tissue-derived or Wharton's jelly-derived hMSCs. Sera from these mice were titrated for each isotype. To confirm specificity of the antibodies, hMSCs were stained with the sera and subjected to a flow cytometic analysis. Spleens were immunostained for proliferating cell nuclear antigen to verify the germinal center formation. Additionally, splenocytes were subjected to a flow cytometric analysis for surface markers including GL-7, B220, CD4, CD8, CD44, and CD62L. Similar experiments were repeated in C57BL/6 mice. The results showed increased IgG₁ and IgG_2a titers in the sera from Balb/c mice injected with hMSCs, and the titers were much higher in the secondary sera than in the primary sera. These antibodies were specifically stained the hMSCs. Germinal centers were observed in the spleen, and flow cytometric analysis of the splenocytes showed higher frequencies of centroblasts (B220⁺ GL7⁺) and memory T cells (CD62L⁺ CD44⁺) both in CD4⁺ and CD8⁺ subsets. Similar results were obtained for C57BL/6 mice. Conclusions: hMSCs induced a humoral immune response in mice, with characters of T cell-dependent immunity.

• Toxicological Research
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• 제35권4호
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• pp.395-402
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• 2019

Subcutaneous adipose tissue (SAT) accumulation is a constitutional disorder resulting from metabolic syndrome. Although surgical and non-surgical methods for reducing SAT exist, patients remain non-compliant because of potential adverse effects and cost. In this study, we developed a new minimally-invasive approach to achieve SAT reduction, using a microneedle (MN) patch prepared from gelatin, which is capable of regulating fat metabolism. Four gelatin types were used: three derived from fish (SA-FG, GT-FG 220, and GT-FG 250), and one from swine (SM-PG 280). We applied gelatin-based MN patches five times over 4 weeks to rats with high-fat diet (HD)-induced obesity, and determined the resulting amount of SAT. We also investigated the histological features and determined the expression levels of fat metabolism-associated genes in SAT using hematoxylin and eosin staining and western blotting, respectively. SAT decreased following treatment with all four gelatin MN patches. Smaller adipocytes were observed in the regions treated with SA-FG, GT-FG 250, and SM-PG 280 MNs, demonstrating a decline in fat accumulation. The expression levels of fat metabolism-associated genes in the MN-treated SAT revealed that GT-FG 220 regulates fatty acid synthase (FASN) protein levels. These findings suggest that gelatin MN patches aid in decreasing the quantity of unwanted SAT by altering lipid metabolism and fat deposition.

• Archives of Plastic Surgery
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• 제39권6호
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• pp.659-662
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• 2012

Progressive facial hemiatrophy, also known as Parry-Romberg syndrome, is a progressive and self-limited deformation of the subcutaneous tissue volume on one side of the face that creates craniofacial asymmetry. We present the case of a patient with a five-year history of progressive right facial hemiatrophy, who underwent facial volumetric restoration using cell-assisted lipotransfer (CAL), which consists of an autologous fat graft enriched with adipose-derived stem cells (ASCs) extracted from the same patient. ASCs have the capacity to differentiate into adipocytes. They also promote angiogenesis, release angiogenic growth factors, and some can survive as stem cells. The use of autologous fat as a filler in soft tissue atrophy has been satisfactory in patients with mild and moderate Parry-Romberg syndrome. Currently, CAL has showed promising results in the long term by decreasing the rate of fat reabsorption. The permanence and stability of the graft in all the injected areas has showed that autologous fat grafts enriched with stem cells could be a promising technique for the correction of defects caused by this syndrome.

• Clinical and Experimental Reproductive Medicine
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• 제48권4호
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• pp.322-336
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• 2021

Objective: Endometriosis is a chronic debilitating inflammatory condition characterized by the presence of endometrial tissues outside the uterine cavity. Pelvic soreness and infertility are the usual association. Due to the poor effectiveness of the hormone therapy and the high incidence of recurrence following surgical excision, there is no single effective option for management of endometriosis. Mesenchymal stem cells (MSCs) are multipotent stromal cells studied for their broad immunoregulatory and anti-inflammatory properties; however, their efficiency in endometriosis cases is still a controversial issue. Our study aim was to evaluate whether adipose tissue-derived MSCs (AD-MSCs) could help with endometriosis through their studied anti-inflammatory role. Methods: Female Wistar rats weighting 180 to 250 g were randomly divided into two groups: group 1, endometriosis group; established by transplanting autologous uterine tissue into rats' peritoneal cavities and group 2, stem cell treated group; treated with AD-MSCs on the 5th day after induction of endometriosis. The proliferative activity of the endometriosis lesions was evaluated through Ki67 staining. Quantitative estimation of interferon γ, tumor necrosis factor-α, interleukin (IL)-6, IL-1β, IL-10, and transforming growth factor β expression, as well as immunohistochemical detection of CD68 positive macrophages, were used to assess the inflammatory status. Results: The size and proliferative activity of endometriosis lesions were significantly reduced in the stem cell treated group. Stem cells efficiently mitigated endometriosis associated chronic inflammatory reactions estimated through reduction of CD68 positive macrophages and the expression of the proinflammatory cytokines. Conclusion: Stem cell therapy can be considered a novel remedy in endometriosis possibly through its anti-inflammatory and antiproliferative properties.

• 한국임상수의학회지
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• 제31권3호
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• pp.253-257
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• 2014

후지 파행 증상을 보이는 다섯 마리 개가 내원하였다. 무릎 관절 신체 검사에서 통증, 파행, 슬개골 탈구, 또는 인대 손상을 확인 하였다. 일반 방사선 사진에서 골증식체와 연골하 병변을 확인 하였고 골관절증 점수가 높은 것을 확인하였다. 수술적 교정과 자가지방 유래 줄기세포 치료의 혼합 방법 또는 자가지방 유래 줄기세포의 경피 주입단독 방법을 보호자의 동의 하에 실시 하였다. $1{\times}10^6$ 개의 줄기세포를 PBS와 함께 주입 하였다. 수술 후 확인은 수의사의 신체검사를 통해 12개월까지 실시 하였다. 주입 후 6개월 또는 12 개월에 파행, 통증, 골관절증 점수가 개선되었음을 확인 하였고 일반 방사선 사진에서도 골증식체와 연골하 병변이 줄어든 것을 확인 할 수 있었다. 본 증례들를 통해 자가지방 유래 줄기 세포 치료가 골관절증을 보이는 개에서 효과적인 방법임을 확인 할 수 있었다.

• 한국임상수의학회지
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• 제29권1호
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• pp.82-86
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• 2012

5년령 중성화된 수컷 치와와가 수개월간의 오른쪽 후지 파행 증상으로 내원하였다. 신체 검사에서 통증을 동반한 4등급의 슬개골 내측 탈구를 확인 하였고 오른쪽 왼쪽 후지 파행등급 2 와 0을 각각 확인 하였다. 일반 방사선 사진에서 골증식체와 연골하 병변을 확인 하였고 오른쪽과 왼쪽 후지 골관절증 점수 20 과 12를 각각 확인하였다. 슬개골 탈구 정복술과 자가지방 유래 줄기세포 치료를 보호자의 동의 하에 결정 하였다. $1{\times}10^6$ 개의 줄기세포를 PBS, 인공 괄절액과 함께 슬개골 탈구 정복술 후에 주입 하였다. 주입 후 19개월에 오른쪽 왼쪽 후지 파행등급 0 와 0, 골관절증 점수 19 와 13을 각각 확인 하였고, 5년후에 파행등급 0 와 0, 골관절증 점수 14 와 15를 각각 확인 하였다. 본 증례를 통해 자가지방 유래 줄기 세포치료를 받은 관절에서 골관절증이 감소 된 것을 확인 할 수 있었다.

• Archives of Plastic Surgery
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• 제35권1호
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• pp.13-19
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• 2008

Purpose: Human adipose tissue-derived mesenchymal stem cells(hATSCs) can be differentiated into multiple mesenchymal lineages, including bone, cartilage, and muscle. And growth hormone play important roles in the normal growth and development of the CNS. In this study, we explored whether the transplanted hATSCs and growth hormones could improve functional recoveries from rats with contusive spinal cord injury. Methods: We divided 30 female rats, which were subjected to a weight driven implant spinal cord injury, into 3 groups with 10 rats each; Group A as a control group, group B with hATSCs transplantation on injured region, and group C with hATSCs transplantation and GH administration for 7 days. Then, we researched their neurologic functional recoveries before and 2, 4, and 8 weeks after transplantation using Basso-Beattie-Bresnahan (BBB) locomotor rating scale. And we checked Y-chromosome positive cells by FISH(Fluorescent in situ hybridization) to identify the survival of transplanted mesenchymal stem cells. Results: After 4 weeks of transplantation, the group B and group C showed significant improvement of neurologic function on BBB locomotor rating scale in comparison with the group A(Group A: $13.1{\pm}0.58$, Group B: $14.6{\pm}0.69$, Group C: $14.9{\pm}0.56$). Moreover, the group C displayed meaningful recovery of neurologic function after 8 weeks in comparison with group B (Group B: $15.7{\pm}0.63$, Group C: $16.5{\pm}1.14$). The group A, the control one, improved for 5 weeks after injury, and had no more recovery. On the other hand, Group B and C showed the improvement of neurologic function continuously for 9 weeks after injury. Conclusion: In this study, we found out that hATSCs transplantation have an effect on neurologic functional recovery of spinal cord injured rat and GH injection seems to bring the synergistic results on this good tendency.

• 생명과학회지
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• 제23권4호
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• pp.471-478
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• 2013

인체 중간엽 줄기세포는 다양한 세포로의 분화 및 자가증식 할 수 있는 능력뿐만 아니라 질병치료에 대한 치료적 잠재력을 가지고 있다. 줄기세포 분화의 분자 기작에 대한 이해는 줄기세포 이식의 치료 효능을 향상시킨다. 본 연구에는 인체 중간엽 줄기세포의 골분화에서 NFAT5의 역할을 밝혔다. 특이적 siRNA의 transfection으로 인한 NFAT5의 억제는 인체 중간엽 줄기세포의 골분화를 현저히 감소시켰으며, NF-${\kappa}B$ promoter 활성화 또한 세포의 증식이나 지방 세포로의 분화에 영향 없이 감소 시켰다. NFAT5의 발현 억제는 기본적으로 유도되는 NF-${\kappa}B$의 활성화와 TNF-${\alpha}$에 의해서 유도되는 NF-${\kappa}B$의 활성화를 감소시켰으나, TNF-${\alpha}$에 의해서 유도되는 NF-${\kappa}B$의 분해에는 아무런 영향을 주지 않았다. 이번 연구를 통해 NFAT5가 NF-${\kappa}B$ 경로를 조절함으로써 인체 중간엽 줄기 세포의 골분화에 아주 중요한 역할을 하는 것을 확인 할 수 있었다.