• 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 the Korean Association of Oral and Maxillofacial Surgeons
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• 제39권2호
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• pp.55-62
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• 2013

Bone tissue engineering is one of the important therapeutic approaches to the regeneration of bones in the entire field of regeneration medicine. Mesenchymal stem cells (MSCs) are actively discussed as material for bone tissue engineering due to their ability to differentiate into autologous bone. MSCs are able to differentiate into different lineages: osteo/odontogenic, adipogenic, and neurogenic. The tissue of origin for MSCs defines them as bone marrow-derived stem cells, adipose tissue-derived stem cells, and, among many others, dental stem cells. According to the tissue of origin, DSCs are further stratified into dental pulp stem cells, periodontal ligament stem cells, stem cells from apical papilla, stem cells from human exfoliated deciduous teeth, dental follicle precursor cells, and dental papilla cells. There are numerous in vitro/in vivo reports suggesting successful mineralization potential or osteo/odontogenic ability of MSCs. Still, there is further need for the optimization of MSCs-based tissue engineering methods, and the introduction of genes related to osteo/odontogenic differentiation into MSCs might aid in the process. In this review, articles that reported enhanced osteo/odontogenic differentiation with gene introduction into MSCs will be discussed to provide a background for successful bone tissue engineering using MSCs with artificially introduced genes.

• The Journal of Korean Physical Therapy
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• 제21권3호
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• pp.87-93
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• 2009

Purpose: TThis study examined the effect of repetitive magnetic stimulation (RMS) on the viability and proliferative response of human adipose tissue-derived stromal cells (hATSCs) in vitro. Methods: The hATSCs were cultured primarily from human adipose tissue harvested by liposuction and incubated in a $37^{\circ}C$ plastic chamber. The cells were exposed to a repetitive magnetic field using a customized magnetic stimulator (Biocon-5000, Mcube Technology). The RMS parameters were set as follows: repetition rate=10Hz, 25Hz (stimulus intensity 100%= 0.1 Tesla, at 4cm from the coil), stimulated time= 1, 5, and 20 minutes. Twenty four hours after one application of RMS, the hATSCs were compared with the sham stimulation, which were kept under the same conditions without the application of RMS. The cells were observed by optical microscopy to determine the morphology and assessed by trypan blue staining for cell proliferation. The apoptosis and viability of the hATSCs were also analyzed by fluorescence-activated cell sorting (FACS) analysis of Annexin V and MTT assay. Results: After RMS, the morphology of the hATSCs was not changed and the apoptosis of hATSCs were not increased compared to the sham stimulation. The viability of the cells was similar to the cells given the sham stimulation. Interestingly, the level of hATSC proliferation was significantly higher in all RMS groups. Conclusion: The application of RMS may not cause a change in morphology and viability of hATSCs but can increase the level of cell proliferation in vitro. RMS might be useful as an adjuvant tool in combination with stem cell therapy without adverse effects.

• 한국응용과학기술학회지
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• 제36권2호
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• pp.394-406
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• 2019

본 연구는 사람의 다양한 세포주를 이용하여 활성산소종(과산화수소수)이 세포의 노화에 미치는 영향을 비교 조사하였다. 여러 농도의 과산화수소수에 세포주를 일주일 동안 배양하여 MTT 방법으로 과산화수소수에 대한 세포 성장의 반억제농도를 구하였다. 그 결과, 50대에서 유래하는 피부 섬유아세포와 10대의 노화 유도 피부 섬유아세포와 비교하여 10대에서 유래하는 피부 섬유아세포에서 과산화수소수에 대한 반억제농도의 값이 유의적으로 더 높았고, 10대의 피부 섬유아세포보다는 10대의 여러 조직 기원하는 성체줄기세포에서 반억제농도의 값이 유의적으로 더 높게 관찰되었다. 또한, 50 ppm 과산화수소수를 1주일 동안 처리한 후, 50대의 피부 섬유아세포에서 다른 세포주에 비해 세포 성장이 현저히 억제되었고, 노화 관련 베타-갈락토시다아제의 활성이 증가되는 것을 관찰하였다. 또한, 활성산소의 세포 독성을 중화시키는 두 유전자, 글루타티온 과산화효소(GPX)와 카탈라아제(CAT)의 발현을 각 세포주에서 조사하였을 때, CAT의 발현은 모든 세포주에서 대체로 낮았지만, GPX 유전자의 발현이 50대의 피부 섬유아세포보다 10대의 피부 섬유아세포와 성체줄기세포에서 현저히 높게 발현되는 것을 관찰하였다. 이상의 결과에서 활성산소는 세포 노화를 유도하고, GPX의 발현이 높은 10대의 피부 섬유아세포와 줄기세포보다는 50대의 피부 섬유아세포와 노화된 피부 섬유아세포에서 활성산소종에 대해 더 큰 민감성을 가지고 있는 것을 알 수 있었다.

• 한국발생생물학회지:발생과생식
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• 제17권4호
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• pp.451-459
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• 2013

The biological activity of tissue specific stem cell is under the control of their specific microenvironment and the exogenous chemicals derived from digestive tract can be one of the constructing factors of that. It is suggested that the extract of brown algae Ishige okamurae has antioxidant-, apoptosis induction-, and antiinflammatory-effects. On the other hand, a few studies have shown that antioxidant assist inhibition of accumulation of fat. So we studied the effect of the extract of I. okamura on the cellular activity and differentiation of 3T3-L1 preadipocyte to adipose cell. The viability of cell was analyzed using 3-[4,5-dimethylthiazo-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Adipogenesis of 3T3-L1 cell was analyzed after induction in the induction medium containing the I. okamurae extract. The cellular activity was high compared with the vehicle and 0.05 mM caffeine in all groups of I. okamurae extract treated cells. The extract of I. okamura inhibited accumulation of lipids in 10 and $50{\mu}g/ml$. The expression of the marker genes for adipocyte differentiation coincided with cytochemical results. These results suggest that the extract of I. okamurae increases the cellular viability of adipose precursor cells. On the other hand, it suppresses the differentiation of preadipocyte to adipocyte and accumulation of lipids in concentration-dependent manners. It may be possible that the major component of the extract can be applied in the control of adipose tissuegenesis.

• International Journal of Stem Cells
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• 제16권4호
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• pp.425-437
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• 2023

Obesity, which continues to increase worldwide, was shown to irreversibly impair the differentiation potential and angiogenic properties of adipose tissue mesenchymal stromal cells (ADSCs). Because these cells are intended for regenerative medicine, especially for the treatment of inflammatory conditions, and the effects of obesity on the immunomodulatory properties of ADSCs are not yet clear, here we investigated how ADSCs isolated from former obese subjects (Ex-Ob) would influence macrophage differentiation and polarization, since these cells are the main instructors of inflammatory responses. Analysis of the subcutaneous adipose tissue (SAT) of overweight (OW) and Ex-Ob subjects showed the maintenance of approximately twice as many macrophages in Ex-Ob SAT, contained within the CD68⁺/FXIII-A^- inflammatory pool. Despite it, in vitro, coculture experiments revealed that Ex-Ob ADSCs instructed monocyte differentiation into a M2-like profile, and under inflammatory conditions induced by LPS treatment, inhibited HLA-DR upregulation by resting M0 macrophages, originated a similar percentage of TNF-α⁺ cells, and inhibited IL-10 secretion, similar to OW-ADSCs and BMSCs, which were used for comparison, as these are the main alternative cell types available for therapeutic purposes. Our results showed that Ex-Ob ADSCs mirrored OW-ADSCs in macrophage education, favoring the M2 immunophenotype and a mixed (M1/M2) secretory response. These results have translational potential, since they provide evidence that ADSCs from both Ex-Ob and OW subjects can be used in regenerative medicine in eligible therapies. Further in vivo studies will be fundamental to validate these observations.

• Archives of Plastic Surgery
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• 제35권6호
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• pp.631-636
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• 2008

Purpose: Adipose tissue-derived stem cells(ADSC) has an osteoconductive potential and demineralized bone matrix(DBM) is an osteoinductive material. A combination of DBM and ADSC wound probably create osteoinductive properties. The purpose of this study is to determine the effect of the combination of DBM and ADSC mixture on healing of rat calvarial defect. Methods: Thirty adult male Sprague-Dawley rats were randomized into 3 groups(n=10) as 1) Control, 2) DBM alone, 3) DBM with ADSC mixture. DBM with ADSC mixture group has had a 3-day preculture of ADSC from groin fat pad. An 6 mm critical size circular calvarial defect was made in each rat. Defect was implanted with DBM alone or DBM with ADSC mixture. Control defect was left unfilled. 6 and 12 weeks after the implantation, the rats were sacrificed and the defects were evaluated by histomorphometric and radiographical studies. Results: Histomorphometric analysis revealed that DBM with ADSC mixture group showed significantly higher bone formation than DBM alone group(p<0.05). Although radiographs from DBM alone group and DBM with ADSC group revealed similar diffuse radiopaque spots dispersed throughout the defect. Densitometric analysis of calvarial defect revealed DBM with ADSC mixture group significantly higher bone formation than DBM alone(p<0.05). There was correlation of densitometry with new bone formation(Spearman's correlation of coefficient=0.804, 6 weeks, 0.802, 12 weeks). Conclusion: The DBM with ADSC mixture group showed the best healing response and the osteoinductive properties of DBM were accelerated with ADSC mixture. It will be clinically applicable that DBM and ADSC mixture in plastic and reconstructive surgery, such as alveolar cleft and congenital facial deformities that bone graft should be required.

• Archives of Plastic Surgery
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• 제39권5호
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• pp.534-539
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• 2012

Background Autologous fat grafting evolved over the twentieth century to become a quick, safe, and reliable method for restoring volume. However, autologous fat grafts have some problems including uncertain viability of the grafted fat and a low rate of graft survival. To overcome the problems associated with autologous fat grafts, we used uncultured adipose tissue-derived stromal cell (stromal vascular fraction, SVF) assisted autologous fat grafting. Thus, the purpose of this study was to evaluate the effect of SVF in a clinical trial. Methods SVF cells were freshly isolated from half of the aspirated fat and were used in combination with the other half of the aspirated fat during the procedure. Between March 2007 and February 2008, a total of 9 SVF-assisted fat grafts were performed in 9 patients. The patients were followed for 12 weeks after treatment. Data collected at each follow-up visit included clinical examination of the graft site(s), photographs for historical comparison, and information from a patient questionnaire that measured the outcomes from the patient perspective. The photographs were evaluated by medical professionals. Results Scores of the left facial area grafted with adipose tissue mixed with SVF cells were significantly higher compared with those of the right facial area grafted with adipose tissue without SVF cells. There was no significant adverse effect. Conclusions The subjective patient satisfaction survey and surgeon survey showed that SVF-assisted fat grafting was a surgical procedure with superior results.

• Molecules and Cells
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• 제41권6호
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• pp.553-561
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• 2018

Icariside II (ICA II) is used in erectile dysfunction treatment. Adipose tissue-derived stem cells (ADSCs) are efficient at improving erectile function. This study aimed to explore the action mechanism of ADSCs in improving erectile function. ADSCs were isolated from the adipose tissues of rats. Cell proliferation was determined using the Cell Counting Kit-8 (CCK-8) assay. The expressions of mRNA and protein were determined separately through qRT-PCR and western blot. The endogenous expressions of related genes were regulated using recombinant plasmids and cell transfection. A Dual-Luciferase Reporter Assay was performed to determine the interaction between miR-34a and STAT3. Rat models with bilateral cavernous nerve injuries (BCNIs) were used to assess erectile function through the detection of mean arterial pressure (MAP) and intracavernosal pressure (ICP). ICA II promoted ADSCs' proliferation and differentiation to Schwann cells (SCs) through the inhibition of miR-34a. Suppressed miR-34a promoted the differentiation of ADSCs to SCs by upregulating STAT3. ICA II promoted the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway. The combination of ICA II and ADSCs preserved the erectile function of the BCNI model rats. ADSCs treated with ICA II markedly preserved the erectile function of the BCNI model rats, which was reversed through miR-34a overexpression. ICA II promotes the differentiation of ADSCs to SCs through the miR34a/STAT3 pathway, contributing to erectile function preservation after the occurrence of a cavernous nerve injury.

• 대한수의학회지
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• 제58권4호
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• pp.211-217
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• 2018

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-${\beta}1$ and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3D-TN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.