• 식물조직배양학회지
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• 제27권6호
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• pp.423-428
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• 2000

In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.

• Clinics in Shoulder and Elbow
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• 제26권3호
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• pp.245-251
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• 2023

Background: For anatomic total arthroscopic repair, cementless humeral fixation has recently gained popularity. However, few studies have compared clinical, radiographic, and patient-reported outcomes between cemented and press-fit humeral fixation, and none have performed follow-up for longer than 5 years. In this study, we compared long-term postoperative outcomes in patients receiving a cemented versus press-fit humeral stem anatomic arthroscopic repair. Methods: This study retrospectively analyzed 169 shoulders that required primary anatomic total shoulder arthroplasty (aTSA). Shoulders were stratified by humeral stem fixation technique: cementation or press-fit. Data were collected pre- and postoperatively. Primary outcome measures included range of motion, patient reported outcomes, and radiographic measures. Results: One hundred thirty-eight cemented humeral stems and 31 press-fit stems were included. Significant improvements in range of motion were seen in all aTSA patients with no significant differences between final cemented and press-fit stems (forward elevation: P=0.12, external rotation: P=0.60, and internal rotation: P=0.77). Patient reported outcome metrics also exhibited sustained improvement through final follow-up. However, at final follow-up, the press-fit stem cohort had significantly better overall scores when compared to the cemented cohort (visual analog score: P=0.04, American Shoulder and Elbow Surgeon Score: P<0.01, Simple Shoulder Test score: P=0.03). Humeral radiolucency was noted in two cemented implants and one press-fit implant. No significant differences in implant survival were observed between the two cohorts (P=0.75). Conclusions: In this series, we found that irrespective of humeral fixation technique, aTSA significantly improves shoulder function. However, within this cohort, press-fit stems provided significantly better outcomes than cemented stems in terms of patient reported outcome scores. Level of evidence: III.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권2호
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• pp.87-93
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• 2010

Introduction: In our previous studies, we isolated porcine skin-derived mesenchymal stem cells (pSDMSCs) from the ears of adult miniature pigs and evaluated the pluripotency of these pSDMSCs based on expressions of transcription factors, such as Oct-4, Sox-2, and Nanog. Moreover, the characteristic of mesenchymal stem cells was revealed by the expression of various mesenchymal stem cell markers, including CD29, CD44, CD90, and vimentin. The aim of this study was to evaluate in vivo osteogenesis after maxillary sinus lift procedures with autogenous pSDMSCs and scaffold. Materials and Methods: The autogenous pSDMSCs were isolated from the 4 miniature pigs, and cultured to 3rd passage with same methods of our previous studies. After cell membranes were labeled using a PKH26, $1{\times}10^{7}$ cells/$100{\mu}L$ of autogenous pSDMSCs were grafted into the maxillary sinus with a demineralized bone matrix (DBM) and fibrin glue scaffold. In the contralateral control side, only a scaffold was grafted, without SDMSCs. After two animals each were euthanized at 2 and 4 weeks after grafting, the in vivo osteogenesis was evaluated with histolomorphometric and osteocalcin immunohistochemical studies. Results: In vivo PKH26 expression was detected in all specimens at 2 and 4 weeks after grafting. Trabecular bone formation and osteocalcin expression were more pronounced around the grafted materials in the autogenous pSDMSCs-grafted group compared to the control group. Newly generated bone was observed growing from the periphery to the center of the grafted material. Conclusion: The results of the present study suggest that autogenous skin-derived mesenchymal stem cells grafting with a DBM and fibrin glue scaffold can be a predictable method in the maxillary sinus floor elevation technique for implant surgery.

• Journal of Periodontal and Implant Science
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• 제41권1호
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• pp.30-43
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• 2011

Purpose: Under different culture conditions, periodontal ligament (PDL) stem cells are capable of differentiating into cementoblast-like cells, adipocytes, and collagen-forming cells. Several previous studies reported that because of the stem cells in the PDL, the PDL have a regenerative capacity which, when appropriately triggered, participates in restoring connective tissues and mineralized tissues. Therefore, this study analyzed the genes involved in mineralization during differentiation of human PDL (hPDL) cells, and searched for candidate genes possibly associated with the mineralization of hPDL cells. Methods: To analyze the gene expression pattern of hPDL cells during differentiation, the hPDL cells were cultured in two conditions, with or without osteogenic cocktails (${\beta}$-glycerophosphate, ascorbic acid and dexamethasone), and a DNA microarray analysis of the cells cultured on days 7 and 14 was performed. Reverse transcription-polymerase chain reaction was performed to validate the DNA microarray data. Results: The up-regulated genes on day 7 by hPDL cells cultured in osteogenic medium were thought to be associated with calcium/iron/metal ion binding or homeostasis (PDE1A, HFE and PCDH9) and cell viability (PCDH9), and the down-regulated genes were thought to be associated with proliferation (PHGDH and PSAT1). Also, the up-regulated genes on day 14 by hPDL cells cultured in osteogenic medium were thought to be associated with apoptosis, angiogenesis (ANGPTL4 and FOXO1A), and adipogenesis (ANGPTL4 and SEC14L2), and the down-regulated genes were thought to be associated with cell migration (SLC16A4). Conclusions: This study suggests that when appropriately triggered, the stem cells in the hPDL differentiate into osteoblasts/cementoblasts, and the genes related to calcium binding (PDE1A and PCDH9), which were strongly expressed at the stage of matrix maturation, may be associated with differentiation of the hPDL cells into osteoblasts/cementoblasts.

• 대한의용생체공학회:학술대회논문집
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• 대한의용생체공학회 1994년도 추계학술대회
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• pp.71-74
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• 1994

In cementless total hip arthroplasty(THA), an initial stability of the femoral component is mandatory to achieve bony ingrowth and secondary long term fixation. Bone ingrowth depends strongly on relative micromotion and stress distributions at the interface. Primary stability of the femoral component can be obtained by minimizing the magnitude of relative micromotions at bone-prosthesis interface, Hence an accurate evaluation of interface behavior and stress/strain fields in the bone implant system may be relevant for better understanding of clinical situations and improving THA design. However, complete evaluation of load transfer in the bone remains difficult to assess experimentally, Hence, recently finite element method (FEM) was introduced in orthopaedic research field to fill the gap due to its unique capacity to evaluate stress in structure of complex shape, loading and material behavior. The authors developed the 3-dimensional numerical finite element model which is composed of totally 1179 elements off and 8 node blick. We also analyzed the micromotions at the bone-stem interface and mechanical behavior of existing bone prosthesis for a loading condition simulating the single leg stance. The result indicates that the values of relative motion for this well fit Multilock stem were $150{\mu}m$ in maximum, $82{\mu}m$ in minimum, and the largest relative motion developed in medial region of proximal femur with anterior-posterior direction. The proximal region of the bone was much larger in motion than the distal region and the stress pattern shows high stress concentration on the cortex near the tip of the stem. These findings indicates that the loading in the proximal femoral bone in the early postoperative situation can produce micromotions on the interface and clinically cementless TEA patient should not be allowed weight bearing strictly early in the postoperative period.

• Journal of Periodontal and Implant Science
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• 제53권1호
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• pp.54-68
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• 2023

Purpose: The purpose of this study was to determine whether metformin (MF) could alleviate the expresssion of reactive oxygen species (ROS) and improve the osteogenic ability of bone marrow mesenchymal stem cells derived from diabetic rats (drBMSCs) in vitro, and to evaluate the effect of MF on the ectopic osteogenesis of drBMSCs in a nude mouse model in vivo. Methods: BMSCs were extracted from normal and diabetic rats. In vitro, a cell viability assay (Cell Counting Kit-8), tests of alkaline phosphatase (ALP) activity, and western blot analysis were first used to determine the cell proliferation and osteogenic differentiation of drBMSCs that were subjected to treatment with different concentrations of MF (0, 50, 100, 200, 500 µM). The cells were then divided into 5 groups: (1) normal rat BMSCs (the BMSCs derived from normal rats group), (2) the drBMSCs group, (3) the drBMSCs + Mito-TEMPO (10 µM, ROS scavenger) group, (4) the drBMSCs + MF (200 µM) group, and (5) the drBMSCs + MF (200 µM) + H₂O₂ (50 µM, ROS activator) group. Intracellular ROS detection, a senescence-associated β-galactosidase assay, ALP staining, alizarin red staining, western blotting, and immunofluorescence assays were performed to determine the effects of MF on oxidative stress and osteogenic differentiation in drBMSCs. In vivo, the effect of MF on the ectopic osteogenesis of drBMSCs was evaluated in a nude mouse model. Results: MF effectively reduced ROS levels in drBMSCs. The cell proliferation, ALP activity, mineral deposition, and osteogenic-related protein expression of drBMSCs were demonstrably higher in the MF-treated group than in the non-MF-treated group. H₂O₂ inhibited the effects of MF. In addition, ectopic osteogenesis was significantly increased in drBMSCs treated with MF. Conclusions: MF promoted the proliferation and osteogenic differentiation of drBMSCs by inhibiting the oxidative stress induced by diabetes and enhenced the ectopic bone formation of drBMSCs in nude mice.

• 구강회복응용과학지
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• 제28권2호
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• pp.127-138
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• 2012

본 연구에서는 임플란트 고정체와 지대주 간의 전하중 크기가 임플란트 주위 변연골의 응력분포에 미치는 영향에 대해 조사하였다. 해석 모델은 하악골에 식립되는 단일 임플란트(solid형 지대주와 submerged 형 고정체)로 제작되었고, 외력 조건으로는 임플란트 지대주 상부에 100N의 기능력이 협설 방향으로 30도 경사져 협측으로 작용하도록 설정하였다. 전하중의 크기가 변연골 응력 분포에 어떠한 차이가 생기게 하는지를 조사하기 위해 다섯개의 다른 전하중의 크기(0, 200, 400, 600, 800N)를 부여하였다. 모든 분석은 선형 탄성을 가정하여 ABAQUS/CAE(ver6.10-1, HKS, Fremont, CA, USA) 프로그램을 사용하였다. 임플란트 주위 변연골 응력분포의 차이는 전하중의 크기와 관련이 있었다. 100N의 교합력 하에 전하중이 0인 경우 변연골(임플란트 벽에서 0.1mm 떨어진 부분)에서 압축 응력은 28.33MPa이었는데, 전하중을 200N 증가시킬 때마다 1.76MPa씩 증가하였다. 이런 방식으로 800N의 전하중을 가할 때 나타나는 최대 압축 응력은 35.18MPa이었다. 반면 변연골에서의 인장 응력은 전하중이 증가할 수록 감소하였다. 임플란트 고정체와 지대주 간의 전하중은 변연골에서의 압축 응력을 증가시킬 수 있으나 기능력에 비하면 그 효과는 미미할 것으로 판단된다.

• Journal of Periodontal and Implant Science
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• 제42권3호
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• pp.73-80
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• 2012

Purpose: The purpose of this study was to investigate the effects of the immunosuppressants FK506 and cyclosporin A (CsA) on the osteogenic differentiation of rat mesenchymal stem cells (MSCs). Methods: The effect of FK506 and CsA on rat MSCs was assessed in vitro. The MTT assay was used to determine the deleterious effect of immunosuppressants on stem cell proliferation at 1, 3, and 7 days. Alkaline phosphatase (ALP) activity was analyzed on days 3, 7, and 14. Alizarin red S staining was done on day 21 to check mineralization nodule formation. Real-time polymerase chain reaction (RT-PCR) was also performed to detect the expressions of bone tissue-specific genes on days 1 and 7. Results: Cell proliferation was promoted more in the FK506 groups than the control or CsA groups on days 3 and 7. The FK506 groups showed increased ALP activity compared to the other groups during the experimental period. The ALP activity of the CsA groups did not differ from the control group in any of the assessments. Mineralization nodule formation was most prominent in the FK506 groups at 21 days. RT-PCR results of the FK506 groups showed that several bone-related genes-osteopontin, osteonectin, and type I collagen (Col-I)-were expressed more than the control in the beginning, but the intensity of expression decreased over time. Runx2 and Dlx5 gene expression were up-regulated on day 7. The effects of 50 nM CsA on osteonectin and Col-I were similar to those of the FK506 groups, but in the 500 nM CsA group, most of the genes were less expressed compared to the control. Conclusions: These results suggest that FK506 enhances the osteoblastic differentiation of rat MSCs. Therefore, FK506 might have a beneficial effect on bone regeneration when immunosuppressants are needed in xenogenic or allogenic stem cell transplantation to treat bone defects.

• 대한심미치과학회지
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• 제32권1호
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• pp.16-22
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• 2023

상악 전치부 임플란트 수복의 성공에 있어서 심미적인 요소는 매우 중요한 부분입니다. 하지만 심미적인 결과를 얻는 것은 쉬운 일이 아니며 특히 치주염으로 인해 치조골 소실이 심하게 발생한 경우라면 더욱 그러합니다. 상악 전치부의 경우, 발치 직후부터 시작되는 치주 조직의 위축이 심미적 수복을 방해하므로 이를 최소화하기 위해 발치 즉시 임플란트를 시행하기도 합니다. 하지만 치조골 소실이 심한 경우 발치 즉시 식립은 심미적 실패를 가져올 가능성이 있고, 이런 결과는 돌이킬 수 없는 문제를 야기합니다. 그래서 치조제 보존술(Alveolar ridge preservation)을 시행하고 이후에 임플란트를 식립하는 방법으로 접근하기도 합니다. 2019년 JCP에 발치 후 임플란트 식립 시기와 발치와 처치에 대한 유럽치주학회의 consensus가 수록되어 있는데 여기서도 '심미적으로 중요한 부위에서 순측 치조골의 소실이 심하게 발생한 경우 치조제 보존술을 고려해야 한다고 언급하고 있습니다. 치조제 보존술을 하게 되면 primary closure가 어렵기 때문에 open membrane technique으로 이차치유를 유도하거나 FGG, CT graft를 시행하게 됩니다. 하지만 이차 치유 과정은 골재생에 부정적인 영향을 줄 수 있고, 연조직 이식은 환자와 술자에게 부담이 될 수 있습니다. 반면 치조골 결손이 심한 발치와에 있는 육아 조직(Granulation tissue)을 이용하게 되면 연조직 이식 없이도 primary closure를 얻을 수 있습니다. 또한 육아 조직에 조직 재생에 도움이 될 수 있는 줄기세포가 함유되어 있다는 연구들이 있습니다. 이를 근거로 치조골 및 연조직 결손이 심한 상악 전치부 치아를 육아 조직을 이용한 치조제 보존술을 통해 임플란트 수복을 시행하였습니다. 결손이 심한 발치와임에도 불구하고 비교적 심미적인 임플란트 수복 결과를 얻을 수 있었습니다.

• The Journal of Advanced Prosthodontics
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• 제6권5호
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• pp.379-386
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• 2014

PURPOSE. These days, mesenchymal stem cells (MSCs) have received worldwide attention because of their potentiality in tissue engineering for implant dentistry. The purpose of this study was to evaluate various growth inducing factors in media for improvement of acquisition of bone marrow mesenchymal stem cells (BMMSCs) and colony forming unit-fibroblast (CFU-F). MATERIALS AND METHODS. The mouse BMMSCs were freshly obtained from female C3H mouse femur and tibia. The cells seeded at the density of $10^6$/dish in media supplemented with different density of fetal bovine serum (FBS), $1{\alpha}$, 25-dihydroxyvitamin (VD3) and recombinant human epidermal growth factor (rhEGF). After 14 days, CFU-F assay was conducted to analyze the cell attachment and proliferation, and moreover for VD3, the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was additionally conducted. RESULTS. The cell proliferation was increased with the increase of FBS concentration (P<.05). The cell proliferation was highest at the density of 20 ng/mL rhEGF compared with 0 ng/mL and 200 ng/mL rhEGF (P<.05). For VD3, although the colony number was increased with the increase of its concentration, the difference was not statistically significant (P>.05). CONCLUTION. FBS played the main role in cell attachment and growth, and the growth factor like rhEGF played the additional effect. However, VD3 did not have much efficacy compare with the other two factors. Improvement of the conditions could be adopted to acquire more functional MSCs to apply into bony defect around implants easily.