• Molecules and Cells
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• 제38권3호
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• pp.267-272
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• 2015

Nacre seashell is a natural osteoinductive biomaterial with strong effects on osteoprogenitors, osteoblasts, and osteoclasts during bone tissue formation and morphogenesis. Although nacre has shown, in one study, to induce bridging of new bone across large non-union bone defects in 8 individual human patients, there have been no succeeding human surgical studies to confirm this outstanding potency. But the molecular mechanisms associated with nacre osteoinduction and the influence on bone marrow-derived mesenchymal stem cells (BMSC's), skeletal stem cells or bone marrow stromal cells remain elusive. In this study we highlight the phenotypic and biochemical effects of Pinctada maxima nacre chips and the global nacre soluble protein matrix (SPM) on primary human bone marrow-derived stromal cells (hBMSCs) in vitro. In static co-culture with nacre chips, the hBMSCs secreted Alkaline phosphatase (ALP) at levels that exceeded bone morphogenetic protein (rhBMP-2) treatment. Concentrated preparation of SPM applied to Stro-1 selected hBMSC's led to rapid ALP secretions, at concentrations exceeding the untreated controls even in osteogenic conditions. Within 21 days the same population of Stro-1 selected hBMSCs proliferated and secreted collagens I-IV, indicating the premature onset of an osteoblast phenotype. The same SPM was found to promote unselected hBMSC differentiation with osteocalcin detected at 7 days, and proliferation increased at 7 days in a dose-dependent manner. In conclusion, nacre particles and nacre SPM induced the early stages of human bone cell differentiation, indicating that they may be promising soluble factors with osteoinductive capacity in primary human bone cell progenitors such as, hBMSC's.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권1호
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• pp.16-23
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• 2005

It is commonly acknowledged that bone morphogenic protein (BMP-2) functions as a potential osteogenic inducer in bone formation. Recently, several papers reported that bone marrow-derived stem cell (BMSC) from human is not responsive to BMP-2 in comparison to high capacity of BMP-2 in the osteoinduction of stromal cell derived from bone marrow of rodent animals such as rat or mouse. In this study, we characterized BMSC derived from 11 years old donor for the responsiveness to rhBMP-2, dexamethasone (Dex) and 1,25-dihydroxyvitamin D (vitamin D), in order to analyze their function in the early osteogenesis. The effect of over mentioned agents was evaluated by means of assessing alkaline phosphatase (ALP) activity/staining, RT-PCR analysis and von Kossa staining. In addition, we analyzed the meaning of expressed several osteoblastic markers such as alkaline phosphatase, collagen typeI, osteopontin, bone sialoprotein and osteocalcin with relation to either differentiation or mineralization. Only in the presence of Dex, human BMSC could commit osteoblastic differentiation and matrix mineralization, and either BMP-2 or vitamin D treatment was not able to induce. But BMP-2 or Vitamin D showed potential synergy effect with Dex. ALP and bone sialoprotein were clearly expressed in response of Dex treatment compared to weak expression of osteopontin in early osteogenesis. Therefore, we expect that this study will contribute partly to elucidiating early osteogenesis mechanism in human, but variations among bone marrow donors must be considered through further study.

• Journal of Korean Neurosurgical Society
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• 제48권5호
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• pp.391-398
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• 2010

Objective : This study was designed to validate the cell trafficking efficiency of the in vivo bioluminescence image (BLI) study in the setting of transplantation of the luciferase expressing bone marrow-derived mesenchymal stem cells (BMSC), which were delivered at each different time after transient middle cerebral artery occlusion (MCAO) in a mouse model. Methods : Transplanting donor BMSC were prepared by primary cell culture from transgenic mouse expressing luciferase (LUC). Transient focal infarcts were induced in 4-6-week-old male nude mice. The experiment mice were divided into five groups by the time of MSC transplantation : 1) sham-operation group, 2) 2-h group, 3) 1-day group, 4) 3-day group, and 5) 1-week group. BLI for detection of spatial distribution of transplanted MSC was performed by detecting emitted photons. Migration of the transplanted cells to the infarcted area was confirmed by histological examinations. Differences between groups were evaluated by paired t-test. Results : A focal spot of bioluminescence was observed at the injection site on the next day after transplantation by Signal intensity of bioluminescence. After 4 weeks, the mean signal intensities of 2-h, 1-day, 3-day, and 1-week group were $2.6{\times}10^7{\pm}7.4{\times}10^6$. $6.1{\times}10^6{\pm}1.2{\times}10^6$, $1.7{\times}10^6{\pm}4.4{\times}10^5$, and $8.9{\times}10^6{\pm}9.5{\times}10^5$, respectively. The 2-h group showed significantly higher signal intensity (p<0.01). The engrafted BMSC showed around the infarct border zones on immunohistochemical examination. The counts of LUC-positive cells revealed the highest number in the 2-h group, in agreement with the results of BLI experiments (p<0.01). Conclusion : In this study, the results suggested that the transplanted BMSC migrated to the infarct border zone in BLI study and the higher signal intensity of LUC-positive cells seen in 2 hrs after MSC transplantation in MCAO mouse model. In addition, noninvasive imaging in real time is an ideal method for tracking stem cell transplantation. This method can be widely applied to various research fields of cell transplantation therapy.

• BMB Reports
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• 제57권2호
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• pp.116-121
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• 2024

We investigated the therapeutic potential of bone marrow-derived mesenchymal stem cell-conditioned medium (BMSC-CM) on immortalized renal proximal tubule epithelial cells (RPTEC/TERT1) in a fibrotic environment. To replicate the increased stiffness characteristic of kidneys in chronic kidney disease, we utilized polyacrylamide gel platforms. A stiff matrix was shown to increase α-smooth muscle actin (α-SMA) levels, indicating fibrogenic activation in RPTEC/TERT1 cells. Interestingly, treatment with BMSC-CM resulted in significant reductions in the levels of fibrotic markers (α-SMA and vimentin) and increases in the levels of the epithelial marker E-cadherin and aquaporin 7, particularly under stiff conditions. Furthermore, BMSC-CM modified microRNA (miRNA) expression and reduced oxidative stress levels in these cells. Our findings suggest that BMSC-CM can modulate cellular morphology, miRNA expression, and oxidative stress in RPTEC/TERT1 cells, highlighting its therapeutic potential in fibrotic kidney disease.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제35권1호
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• pp.7-12
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• 2009

Purpose: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. Material & Methods: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. Results: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores ($20-40{\mu}m$). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. Conclusion: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.

• International Journal of Stem Cells
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• 제16권1호
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• pp.66-77
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• 2023

Background and Objectives: We compared the efficacy and safety of human bone marrow-derived mesenchymal stem cells (hBMSC), delivered at different doses and via different injection routes in an animal model of chronic kidney disease. Methods and Results: A total of ninety 12-week-old rats underwent 5/6 nephrectomy and randomized among nine groups: sham, renal artery control (RA-C), tail vein control (TV-C), renal artery low dose (RA-LD) (0.5×10⁶ cells), renal artery moderate dose (RA-MD) (1.0×10⁶ cells), renal artery high dose (RA-HD) (2.0×10⁶ cells), tail vein low dose (TV-LD) (0.5×10⁶ cells), tail vein moderate dose (TV-MD) (1.0×10⁶ cells), and tail vein high dose (TV-HD) (2.0×10⁶ cells). Renal function and mortality of rats were evaluated after hBMSC injection. Serum blood urea nitrogen was significantly lower in the TV-HD group at 2 weeks (p<0.01), 16 weeks (p<0.05), and 24 weeks (p<0.01) than in the TV-C group, as determined by one-way ANOVA. Serum creatinine was significantly lower in the TV-HD group at 24 weeks (p<0.05). At 8 weeks, creatinine clearance was significantly higher in the TV-MD and TV-HD groups (p<0.01, p<0.05) than in the TV-C group. In the safety evaluation, we observed no significant difference among the groups. Conclusions: Our findings confirm the efficacy and safety of high dose (2×10⁶ cells) injection of hBMSC via the tail vein.

• International Journal of Stem Cells
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• 제16권2호
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• pp.202-214
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• 2023

Background and Objectives: To investigate the role of exosomes from periodontal ligament cells (PDLCs) in bone marrow mesenchymal stem cell (BMSC) migration. Methods and Results: Human PDLCs were applied cyclic tension stretching. Exosomes were extracted from cultured PDLCs by ultracentrifugation, then characterized for their size, morphology and protein markers by NTA, TEM and western blotting. The process that PKH26-labeled exosomes taken up by BMSCs was assessed by confocal microscope. BMSC migration was examined by Transwell assay. Exosomes derived from PDLCs were identified. Cyclic tension stretch application on PDLCs can enhance the migration ability of BMSCs through exosomes. The exosomal miRNA expression profiles of unstretched and stretched PDLCs were tested by miRNA microarray. Four miRNAs (miR-4633-5p, miR-30c-5p, miR-371a-3p and let-7b-3p) were upregulated and six (miR-4689, miR-8485, miR-4655-3p, miR-4672, miR-3180-5p and miR-4476) were downregulated in the exosomes after stretching. Sixteen hub proteins were found in the miRNA-mRNA network. Gene Ontology and KEGG pathway analyses demonstrated that the target genes of differentially expressed exosomal miRNAs closely related to the PI3K pathway and vesicle transmission. Conclusions: The exosomes derived from cyclic tension-stretched PDLCs can promote the migration of BMSCs. Alternation of microRNA profiles provides a basis for further research on the regulatory function of the exosomal miRNAs of PDLCs during orthodontic tooth movement.

• Restorative Dentistry and Endodontics
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• 제45권4호
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• pp.52.1-52.11
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• 2020

Objectives: Yttria-stabilized tetragonal phase zirconia has been used as a dental restorative material for over a decade. While it is still the strongest and toughest ceramic, its translucency remains as a significant drawback. To overcome this, stabilizing the translucency zirconia to a significant cubic crystalline phase by increasing the yttria content to more than 8 mol% (8YTZP). However, the biocompatibility of a high amount of yttria is still an important topic that needs to be investigated. Materials and Methods: Commercially available 8YTZP plates were used. To enhance cell adhesion, proliferation, and differentiation, the surface of the 8YTZP is sequentially polished with a SiC-coated abrasive paper and surface coating with type I collagen. Fibroblast-like cells L929 used for cell adherence and cell proliferation analysis, and mouse bone marrow-derived mesenchymal stem cells (BMSC) used for cell differentiation analysis. Results: The results revealed that all samples, regardless of the surface treatment, are hydrophilic and showed a strong affinity for water. Even the cell culture results indicate that simple surface polishing and coating can affect cellular behavior by enhancing cell adhesion and proliferation. Both L929 cells and BMSC were nicely adhered to and proliferated in all conditions. Conclusions: The results demonstrate the biocompatibility of the cubic phase zirconia with 8 mol% yttria and suggest that yttria with a higher zirconia content are not toxic to the cells, support a strong adhesion of cells on their surfaces, and promote cell proliferation and differentiation. All these confirm its potential use in tissue engineering.

• 폴리머
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• 제29권5호
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• pp.501-507
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• 2005

소장 점막하 조직(SIS)은 면역반응이 없어 생체재료로 널리 사용되고 있다. 본 연구에서는 SIS를 스폰지 형태로 제조하여 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride(EDC)를 이용하여 경화시켰으며, SIS 스폰지의 구성 윈소를 알아보기 위해 원소분석(EA)과 에너지 분산 X선 분광계(EDS)를 사용해 분석하였다. 또한 SIS 함량과 EDC의 농도에 따른 섬유아세포의 부착도 및 성장도를 알아보기 위해 methylthiazoletetrazolium,(MTT)을 실시하였다. 이 스폰지에 골수 간엽 줄기세포(BMSCs)를 파종해 4주 동안 조직공학적 골분화를 유도하였다. 골분화 유도를 위해 골분화 배지를 사용했으며, 배지에 따른 BMSCs의 세포 성장도와 alkaline phosphatase(ALP) 활성을 측정해 보았다. 또한 역전사 중합연쇄반응을 통해 골분화 여부를 관찰하였다. SEM 관찰 결과 모든 스폰지에서 균일한 형태의 열린 다공이 형성되었음을 확인할 수 있었다. RT-PCR 결과 4주 동안 골분화 배지를 주었을 때 제 I형 교원질이 발현됨을 확인할 수 있었으며, ALP 결과에서도 골분화 배지에서 ALP활성이 높게 나타남을 볼 수 있었다. 결론적으로 제조한 SIS 스폰지는 조직공학적 담체로써 우수한 특성을 보이고 있었으며, 또한 BMSCs의 골분화 유도에도 좋은 결과를 보임으로써 조직공학적 골 재생에 잠재적인 가능성을 가지고 있음을 확인할 수 있었다.