• Biotechnology and Bioprocess Engineering:BBE
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• 제11권4호
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• pp.364-367
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• 2006

In order to enhance the repair of defects in articular cartilage via cell therapy with autologous chondrocytes, as well as with periosteum-derived progenitor cells (PDPCs), silkworm hemolymph (SH) and a variety of cartilage-specific extracellular matrices (ECMs) including type II collagen, proline, chondroitin 4-sulfate, and chondroitin 6-sulfate were assessed with regard to their efficacy as media supplements. SH, a known anti-apoptotic agent, was found to enhance cell growth, as was shown by the results of a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to the results of reverse transcriptase polymerase chain reaction (RT-PCR) analyses, the cartilage-specific ECMs were found to stimulate the expression of hyaline cartilage-specific genes, most notably type II collagen and Sox9, in monolayer cultures of PDPCs.

• 대한의용생체공학회:의공학회지
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• 제26권2호
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• pp.101-110
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• 2005

Periosteum and periosteum-derived progenitor cells have demonstrated the potential for stimulative applications in repairs of various musculoskeletal tissues. It has been found that the periosteum contains mesenchymal progenitor cells capable of differentiating into either osteoblasts or chondrocytes depending on the culture conditions. Anatomically, the periosteum is a heterogeneous multi-layered membrane, consisting of an inner cambium and an outer fibrous layer. The present study was designed to elucidate the cellular phenotypic characteristics of cambium and fibrous layer cells in vitro, and to assess whether structural integrity of the tendon in the bone tunnel can be improved by periosteal augmentation of the tendon­bone interface. It was found the cells from each layer showed distinct phenotypic characteristics in a primary monolayer culture system. Specifically, the cambium cells demonstrated higher osteogenic characteristics (higher alkaline phosphatase and osteocalcin levels), as compared to the fibrous cells. Also in vivo animal model showed that a periosteal augmentation of a tendon graft could enhance the structural integrity of the tendon-bone interface, when the periosteum is placed between the tendon and bone interface with the cambium layer facing toward the bone. These findings suggest that extra care needs to be taken in order to identify and maintain the intrinsic phenotypes of the heterogeneous cell types within the periosteum. This will improve our understanding of periosteum in applications for musculoskeletal tissue repairs and tissue engineering.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권6호
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• pp.550-552
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• 2006

Periosteum-derived progenitor cells (PDPCs) were isolated using a fluorescence-activated cell sorter and their chondrogenic potential in biomaterials was investigated for the treatment of defective articular cartilage as a cell therapy. The chondrogenesis of PDPCs was conducted in a thermoreversible gelation polymer (TGP), which is a block copolymer composed of temperature-responsive polymer blocks such as poly(N-isopropylacrylamide) and of hydrophilic polymer blocks such as polyethylene oxide, and a defined medium that contained transforming growth $factor-{\beta}3\;(TGF-{\beta}3)$. The PDPCs exhibited chondrogenic potential when cultured in TGP. As the PDPCs-TGP is an acceptable biocompatible complex appropriate for injection into humans, this product might be readily applied to minimize invasion in a defected knee.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.239-243
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• 2005

본 실험에서는 골막조직 내에 존재하는 PDPCs를 분리하고, 기존 피부 이식을 위해 사용되는 $Hyalrograft^{\circledR}$ 3D에서의 chondogenesis가 가능함을 확인하였다. 수적인 확보가 제한되는 연골세포의 대체 세포로서 PDPCs의 가능성을 확인하였으며 동시에 $Hyalrograft^{\circledR}$ 3D에서의 연골화는 지지체의 양적 확보에 있어 기존의 $Hyalograft^{\circledR}$ C 보다 경제적일 수 있음을 의미한다.

• KSBB Journal
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• 제31권2호
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• pp.120-125
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• 2016

Two-dimensional cultivation is typically used for cell growth, but the method reduces the characteristics of chondrocytes and stem cells, and limits culture area. Therefore, development of three-dimensional culture method is needed to mimic in vivo environment, improve quality of cells and scale-up efficiently. Improving proliferation and chondrogenesis is available by co-culture of chondrocytes and mesenchymal stem cells (MSCs) that leads to interaction between two kinds of cells. However, the co-culture has problems that permeability of sphere diminishes as aggregate size increased and ratio of two kinds of cells composing each spheres is different. In this work, co-cultivation method using controlled sphere composed of chondrocytes and MSCs was established and enhanced chondrogenesis. Periosteum-derived progenitor cells (PDPCs) that are appropriate for cell therapy source of articular cartilage were used as MSCs. Controlled spheres were formed in the hanging-drop plates and shifted for being induced chondrogenesis in 35-mm non-adhesive culture dishes at a rotation rate of 60 rpm. After inducing chondrogenesis, gene expressions related with chondrogenesis were found to be improved and it was apparent that the utilization of controlled spheres promoted chondrogenesis. As a result, available numbers of cells per unit area were increased and chondrogenic differentiation ability was improved compared to typical two-dimensional culture. This approach shows the potential in cartilage regeneration as it can provide sufficient numbers of chondrocytes.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제29권3호
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• pp.197-205
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• 2007

Angiogenesis is a essential part for bone formation and bone fracture healing. Vascular endothelial growth factor (VEGF), one of the most important molecules among many angiogenic factors, is a specific mitogen for vascular endothelial cells. VEGF-mediated angiogenesis is required for bone formation and repair. However, the effect of VEGF on osteoblastic cells during osteogenesis is still controversial. In recent days, substantial progress have been made toward developing tissue-engineered alternatives to autologous bone grafting for maxillofacial bony defects. Periosteum has received considerable interest as a better source of adult stem cells. Periosteum has the advantage of easy harvest and contains various cell types and progenitor cells that are able to differentiate into a several mesenchymal lineages, including bone. Several studies have reported the bone formation potential of periosteal cells, however, the correlation between VEGF signaling and cultured human periosteal cell-derived osteogenesis has not been fully investigated yet. The purpose of this study was to examine the correlation between VEGF signaling and cultured human periosteal-derived cells osteogenesis. Periosteal tissues of $5\;{\times}\;20\;mm$ were obtained from mandible during surgical extraction of lower impacted third molar from 3 patients. Periosteal-derived cells were introduced into the cell culture and were subcultured once they reached confluence. After passage 3, the periosteal-derived cells were further cultured for 42 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and ${\beta}-glycerophosphate$. We evaluated the alkaline phosphatase (ALP) activity, the expression of Runx2 and VEGF, alizarin red S staining, and the quantification of osteocalcin and VEGF secretion in the periosteal-derived cells. The ALP activity increased rapidly up to day 14, followed by decrease in activity to day 35. Runx2 was expressed strongly at day 7, followed by decreased expression at day 14, and its expression was not observed thereafter. Both VEGF 165 and VEGF 121 were expressed strongly at day 35 and 42 of culture, particularly during the later stages of differentiation. Alizarin red S-positive nodules were first observed on day 14 and then increased in number during the entire culture period. Osteocalcin and VEGF were first detected in the culture medium on day 14, and their levels increased thereafter in a time-dependent manner. These results suggest that VEGF secretion from cultured human periosteal-derived cells increases along with mineralization process of the extracellular matrix. The level of VEGF secretion from periosteal-derived cells might depend on the extent of osteoblastic differentiation.