• Title/Summary/Keyword: Tissue engineering applications

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Hypoxic condition enhances chondrogenesis in synovium-derived mesenchymal stem cells

  • Bae, Hyun Cheol;Park, Hee Jung;Wang, Sun Young;Yang, Ha Ru;Lee, Myung Chul;Han, Hyuk-Soo
    • Biomaterials Research
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    • v.22 no.4
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    • pp.271-278
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    • 2018
  • Background: The chondrogenic differentiation of mesenchymal stem cells (MSCs) is regulated by many factors, including oxygen tensions, growth factors, and cytokines. Evidences have suggested that low oxygen tension seems to be an important regulatory factor in the proliferation and chondrogenic differentiation in various MSCs. Recent studies report that synovium-derived mesenchymal stem cells (SDSCs) are a potential source of stem cells for the repair of articular cartilage defects. But, the effect of low oxygen tension on the proliferation and chondrogenic differentiation in SDSCs has not characterized. In this study, we investigated the effects of hypoxia on proliferation and chondrogenesis in SDSCs. Method: SDSCs were isolated from patients with osteoarthritis at total knee replacement. To determine the effect of oxygen tension on proliferation and colony-forming characteristics of SDSCs, A colony-forming unit (CFU) assay and cell counting-based proliferation assay were performed under normoxic (21% oxygen) or hypoxic (5% oxygen). For in vitro chondrogenic differentiation, SDSCs were concentrated to form pellets and subjected to conditions appropriate for chondrogenic differentiation under normoxia and hypoxia, followed by the analysis for the expression of genes and proteins of chondrogenesis. qRT-PCR, histological assay, and glycosoaminoglycan assays were determined to assess chondrogenesis. Results: Low oxygen condition significantly increased proliferation and colony-forming characteristics of SDSCs compared to that of SDSCs under normoxic culture. Similar pellet size and weight were found for chondrogensis period under hypoxia and normoxia condition. The mRNA expression of types II collagen, aggrecan, and the transcription factor SOX9 was increased under hypoxia condition. Histological sections stained with Safranin-O demonstrated that hypoxic conditions had increased proteoglycan synthesis. Immunohistochemistry for types II collagen demonstrated that hypoxic culture of SDSCs increased type II collagen expression. In addition, GAG deposition was significantly higher in hypoxia compared with normoxia at 21 days of differentiation. Conclusion: These findings show that hypoxia condition has an important role in regulating the synthesis ECM matrix by SDSCs as they undergo chondrogenesis. This has important implications for cartilage tissue engineering applications of SDSCs.

Bio-Conjugated Polycaprolactone Membranes: A Novel Wound Dressing

  • Cai, Elijah Zhengyang;Teo, Erin Yiling;Jing, Lim;Koh, Yun Pei;Qian, Tan Si;Wen, Feng;Lee, James Wai Kit;Hing, Eileen Chor Hoong;Yap, Yan Lin;Lee, Hanjing;Lee, Chuen Neng;Teoh, Swee-Hin;Lim, Jane;Lim, Thiam Chye
    • Archives of Plastic Surgery
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    • v.41 no.6
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    • pp.638-646
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    • 2014
  • Background The combination of polycaprolactone and hyaluronic acid creates an ideal environment for wound healing. Hyaluronic acid maintains a moist wound environment and accelerates the in-growth of granulation tissue. Polycaprolactone has excellent mechanical strength, limits inflammation and is biocompatible. This study evaluates the safety and efficacy of bio-conjugated polycaprolactone membranes (BPM) as a wound dressing. Methods 16 New Zealand white rabbits were sedated and local anaesthesia was administered. Two $3.0{\times}3.0cm$ full-thickness wounds were created on the dorsum of each rabbit, between the lowest rib and the pelvic bone. The wounds were dressed with either BPM (n=12) or Mepitel (n=12) (control), a polyamide-silicon wound dressing. These were evaluated macroscopically on the 7th, 14th, 21st, and 28th postoperative days for granulation, re-epithelialization, infection, and wound size, and histologically for epidermal and dermal regeneration. Results Both groups showed a comparable extent of granulation and re-epithelialization. No signs of infection were observed. There was no significant difference (P>0.05) in wound size between the two groups. BPM (n=6): $8.33cm^2$, $4.90cm^2$, $3.12cm^2$, $1.84cm^2$; Mepitel (n=6): $10.29cm^2$, $5.53cm^2$, $3.63cm^2$, $2.02cm^2$; at the 7th, 14th, 21st, and 28th postoperative days. The extents of epidermal and dermal regeneration were comparable between the two groups. Conclusions BPM is comparable to Mepitel as a safe and efficacious wound dressing.

Trends in Saliva Research and Biomedical Clinical Applications (타액 연구의 최신 지견과 임상 응용)

  • Soyoung Park;Eungyung Lee;Jonghyun Shin;Taesung Jeong
    • Journal of the korean academy of Pediatric Dentistry
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    • v.50 no.1
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    • pp.1-12
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    • 2023
  • Function of salivary gland and saliva composition can be an indicator of individual's health status. Recently, saliva has been thought to have a high potential for usage in the biomedical field to diagnose, evaluate, and prevent systemic health due to the technological advances in analyzing and detecting small elements such as immunological and metabolic products, viruses, microorganisms, hormones in saliva. As a diagnostic specimen, saliva has some useful advantages compared to serum. Because of simple non-invasive method, saliva sampling is quite comfort for the patient, and it doesn't require specialists to collect samples. The possibility of infection during the collection process is also low. For this reason, proteins, genetic materials, and various biomarkers in saliva are actively being utilized on studying stress, microbiomics, genetics, and epigenetics. For the research on collecting big data related to systemic health, the needs on biobank has been focused. Regeneration of salivary gland based on tissue engineering has been also on advancement. However, there are still many issues to be solved, such as the standardization of sample collection, storage, and usage. This review focuses on the recent trends in the field of saliva research and highlight the future perspectives in biomedical and other applications.

Microencapsulation of Iron Oxide Nanoparticles and Their Application in Magnetic Levitation of Cells (산화철 나노입자의 마이크로캡슐화와 이를 이용한 세포의 자력부상 배양)

  • Lee, Jin Sil;Lee, Joon ho;Shim, Jae Kwon;Hur, Won
    • Applied Chemistry for Engineering
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    • v.31 no.1
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    • pp.13-18
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    • 2020
  • Iron oxide nanoparticles were microencapsulated using fibroin, a protein polymer of silk fiber, for theragnostic applications. The content of iron oxide was determined to be 4.28% by thermogravimetric analysis and 5.11% by magnetometer. A suspension of murine fibroblast 3T3 cells grown in medium supplemented with iron oxide-microcapsules turned clear in response to the magnetic force and the cells aggregated to the magnet direction. Neodymium magnets placed on the top of the culture dish, and attracted cells to the center of the culture surface. The cells collected on the culture surface aggregated to form a rough spheroid of 2 mm in a diameter after 72 h. In the outer layer of the cell aggregate, cells were relatively large and gathered together to form a dense tissue, but the central part was observed to undergo cell death due to the mass transfer restriction. In the outer layer, iron oxide-microcapsules were lined up like chains in the direction of magnetic force. Using microCT, it was demonstrated that the iron oxides inside the cell aggregate were not evenly distributed but biased to the magnetic direction.

Electrochemical Characteristics of Zn and Si Ion-doped HA Films on Ti-6Al-4V by PEO Treatment

  • Lim, Sang-Gyu;Hwang, In-Jo;Choe, Han-Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2016.11a
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    • pp.199-199
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    • 2016
  • Commercially pure titanium (cp-Ti) and Ti alloys (typically Ti-6Al-4V) display excellent corrosion resistance and biocompatibility. Although the chemical composition and topography are considered important, the mechanical properties of the material and the loading conditions in the host have, conventionally. Ti and its alloys are not bioactive. Therefore, they do not chemically bond to the bone, whereas they physically bond with bone tissue. The electrochemical deposition process provides an effective surface for biocompatibility because large surface area can be served to cell proliferation. Electrochemical deposition method is an attractive technique for the deposition of hydroxyapatite (HAp). However, the adhesions of these coatings to the Ti surface needs to be improved for clinical used. Plasma electrolyte oxidation (PEO) enables control in the chemical com position, porous structure, and thickness of the $TiO_2$ layer on Ti surface. In addition, previous studies h ave concluded that the presence of $Ca^{+2}$ and ${PO_4}^{3-}$ ion coating on porous $TiO_2$ surface induced adhesion strength between HAp and Ti surface during electrochemical deposition. Silicon (Si) in particular has been found to be essential for normal bone and cartilage growth and development. Zinc (Zn) plays very important roles in bone formation and immune system regulation, and is also the most abundant trace element in bone. The objective of this work was to study electrochemical characteristcs of Zn and Si coating on Ti-6Al-4V by PEO treatment. The coating process involves two steps: 1) formation of porous $TiO_2$ on Ti-6Al-4V at high potential. A pulsed DC power supply was employed. 2) Electrochemical tests were carried out using potentiodynamic and AC impedance methoeds. The morphology, the chemical composition, and the micro-structure an alysis of the sample were examined using FE-SEM, EDS, and XRD. The enhancements of the HAp forming ability arise from $Si/Zn-TiO_2$ surface, which has formed the reduction of the Si/Zn ions. The promising results successfully demonstrate the immense potential of $Si/Zn-TiO_2$ coatings in dental and biomaterials applications.

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Effects of Macrophage on Biodegradation of β-tricalcium Phosphate Bone Graft Substitute (대식세포가 β-tricalcium Phosphate 뼈이식제의 생분해에 미치는 영향)

  • Kim, Young-Hee;Jyoti, Anirban;Byun, In-Sun;Oh, Ik-Hyun;Min, Young-Ki;Yang, Hun-Mo;Lee, Byong-Taek;Song, Ho-Yeon
    • Journal of the Korean Ceramic Society
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    • v.45 no.10
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    • pp.618-624
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    • 2008
  • Various calcium phosphate bioceramics are distinguished by their excellent biocompatibility and osteoconductivity. Especially, the exceptional biodegradability of $\beta$-TCP makes it a bone graft substitute of choice in many clinical applications. The activation of osteoclasts, differentiated from macrophage precursor cells, trigger a cell-mediated resorption mechanism that renders $\beta$-TCP biodegradable. Based on this evidence, we studied the biodegradation process of granular-type $\beta$-TCP bone graft substitute through in vitro and in vivo studies. Raw 264.7 cells treated with RANKL and M-CSF differentiated into osteoclasts with macrophage-like properties, as observed with TRAP stain. These osteoclasts were cultured with $\beta$-TCP nano powders synthesized by microwave-assisted process. We confirmed the phagocytosis of osteoclasts by observing $\beta$-TCP particles in their phagosomes via electron microscopy. No damage to the osteoclasts during phagocytosis was observed, nor did the $\beta$-TCP powders show any sign of cytotoxicity. We also observed the histological changes in subcutaneous tissues of rats implanted with granule-type $\beta$-TCP synthesized by fibrous monolithic process. The $\beta$-TCP bone graft substitute was well surrounded with fibrous tissue, and 4 months after implantation, 60% of its mass had been biodegraded. Also, histological findings via H&E stain showed a higher level of infiltration of lymphocytes as well as macrophages around the granule-type $\beta$-TCP. From the results, we have concluded that macrophages play an important role in the biodegradation process of $\beta$-TCP bone graft substitutes.

Behavior of Retinal Pigment Epithelial Cells Cultured on Silk Films (실크필름에 배양한 망막색소상피세포의 거동)

  • Lee, So Jin;Kim, Hye Yun;Kim, Seul Ji;Yang, Jaewon;Lee, Seon Ui;Park, Chan Hum;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.3
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    • pp.364-370
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    • 2014
  • The retinal pigment epithelium (RPE) plays an important role in maintaining a healthy retina and the degeneration of RPE caused a number of retinal diseases. The transplantation of RPE has recently become a possible therapeutic modality for retinal degeneration. To transplant RPE cells securely, substrates are essential, and then as a substrate, we fabricated films using silk that has unique mechanical properties and biocompatibility. After the FTIR spectra, contact angle and biodegradation of silk films were confirmed, RPE cells were seeded and the influence of RPE cells on silk films was examined. We measured the cell adhesion, cell viability, morphology and specific mRNA expression by MTT assay, SEM, immunofluorescence and RT-PCR. In this study, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells cultured on silk films were great, and thereby we were able to confirm the potential applications of silk films as tissue engineering carrier for regeneration of retina.

Adhesion and Proliferation Behavior of Retinal Pigment Epithelial Cells on Hesperidin/PLGA Films (헤스페리딘/PLGA 필름에서 망막색소상피세포의 부착과 증식거동)

  • Lee, So Jin;Kang, Su Ji;Kim, Hye Yun;Lee, Jung Hwan;Kim, Eun Young;Kwon, Soon Yong;Chung, Jin Wha;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.24-30
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    • 2014
  • Retinal pigment epithelium (RPE) plays an important role in maintaining the visual function and the degeneration of the RPE causes several retinal degeneration disease. In order to fabricate the suitable carrier for RPE transplantation, the hybrid poly(lactide-co-glycolide) (PLGA) film with hesperidin was prepared. Hesperidin has an anti-inflammatory and antioxidant characteristics. ARPE-19 was seeded on hesperidin/PLGA film and then, cell proliferation was determined by the MTT assay, and cell adhesion and cell morphology were confirmed by SEM. Also, RT-PCR was performed to confirm the expression of the specific genes, and AEC immunohistochemical staining was performed to determine the expression of RPE65. As a result, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells were more excellent on hesperidin/PLGA film than PLGA film, thereby we were able to confirm the potential applications of hesperidin/PLGA film as tissue engineering carrier for regeneration of retina.

Voxel-based Investigations of Phase Mask Effects on Susceptibility Weighted Images (화소 간 분석을 이용하여 자화율 가중 영상(SWI)에 나타난 위상 마스킹의 효과 분석)

  • Hwang, Eo-Jin;Kim, Min-Ji;Kim, Hyug-Gi;Ryu, Chang-Woo;Jahng, Geon-Ho
    • Progress in Medical Physics
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    • v.24 no.1
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    • pp.25-34
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    • 2013
  • To investigate effects of phase mask on susceptibility-weighted images (SWI) using voxel-based analyses in normal elderly subjects. A three-dimensional (3D) gradient echo sequence ran to obtain SWIs in 20 healthy elderly subjects. SWIs with two (SWI2) and four (SWI4) phase multiplications were achieved with positive (PSWI) and negative (NSWI) phase masks to investigate phase mask effects. The voxel-based comparisons were performed using paired t-tests between PSWI and NSWI and between SWI2 and SWI4. Differences of signal intensities between magnitude images and SWI4 were larger than those between magnitude images and SWI2s. Differences of signal intensities between magnitude images and PSWIs were larger than those between magnitude images and NSWIs. Moreover, the signal intensities from NSWI2s and NSWI4s were greater than those from PSWI2s and PSWI4s, respectively. More differences of signal intensities between NSWI4 and PSWI4s were found than those between NSWI2s and PSWI2s in the whole brain images. The voxel-based analyses of SWI could be beneficial to investigate susceptibility differences on the entire brain areas. The phase masking method could be chosen to enhance brain tissue contrast rather than to enhance venous blood vessels. Therefore, it is recommended to apply voxel-based analyses of SWI to investigate clinical applications.

Virus Inactivation during the Manufacture of a Collagen Type I from Bovine Hides (소 가죽 유래 Type I Collagen 생산 공정에서 바이러스 불활화)

  • Bae, Jung Eun;Kim, Chan Kyung;Kim, Sungpo;Yang, Eun Kyung;Kim, In Seop
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.314-318
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    • 2012
  • Most types of collagen used for biomedical applications, such as cell therapy and tissue engineering, are derived from animal tissues. Therefore, special precautions must be taken during the production of these proteins in order to assure against the possibility of the products transmitting infectious diseases to the recipients. The ability to remove and/or inactivate known and potential viral contaminants during the manufacturing process is an ever-increasingly important parameter in assessing the safety of biomedical products. The purpose of this study was to evaluate the efficacies of the 70% ethanol treatment and pepsin treatment at pH 2.0 for the inactivation of bovine viruses during the manufacture of collagen type I from bovine hides. A variety of experimental model viruses for bovine viruses including bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), bovine parainfluenza 3 virus (BPIV-3), and bovine parvovirus (BPV), were chosen for the evaluation of viral inactivation efficacy. BHV, BVDV, BPIV-3, and BPV were effectively inactivated to undetectable levels within 1 h of 70% ethanol treatment for 24 h, with log reduction factors of ${\geq}5.58$, ${\geq}5.32$, ${\geq}5.11$, and ${\geq}3.42$, respectively. BHV, BVDV, BPIV-3, and BPV were also effectively inactivated to undetectable levels within 5 days of pepsin treatment for 14 days, with the log reduction factors of ${\geq}7.08$, ${\geq}6.60$, ${\geq}5.60$, and ${\geq}3.59$, respectively. The cumulative virus reduction factors of BHV, BVDV, BPIV-3, and BPV were ${\geq}12.66$, ${\geq}11.92$, ${\geq}10.71$, and ${\geq}7.01$. These results indicate that the production process for collagen type I from bovine hides has a sufficient virus-reducing capacity to achieve a high margin of virus safety.