• Journal of Chest Surgery
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• v.45 no.6
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• pp.380-389
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• 2012

Background: Bovine pericardium is one of the most widely used materials in bioprosthetic heart valves. Immunologic responses have been implicated as potential causes of limited durability of xenogenic valves. This study aimed to determine the effectiveness of decellularization and ${\alpha}$-galactosidase (${\alpha}$-gal) to remove major xenoreactive antigens from xenogenic tissues. Materials and Methods: Recombinant Bacteroides thetaiotaomicron (B. thetaiotaomicron) ${\alpha}$-gal or decellularization, or both were used to remove ${\alpha}$-gal from bovine pericardium. It was confirmed by ${\alpha}$-gal-bovine serum albumin-based enzyme-linked immunosorbent assay (ELISA), high-performance anion exchange chromatography, flow cytometry, 3,3'-diaminobenzidine-staining, and lectin-based ELISA. The mechanical properties of bovine pericardium after decellularization or ${\alpha}$-gal treatment were investigated by tests of tensile-strength, permeability, and compliance. Collagen fiber rearrangement was also evaluated by a 20,000${\times}$ transmission electron microscope (TEM). Results: Recombinant B. thetaiotaomicron ${\alpha}$-gal could effectively remove ${\alpha}$-gal from bovine pericardium B. thetaiotaomicron (0.1 U/mL, pH 7.2) while recombinant human ${\alpha}$-gal removed it recombinant human ${\alpha}$-gal (10 U/mL, pH 5.0). There was no difference in the mechanical properties of fresh and recombinant ${\alpha}$-gal-treated bovine pericardium. Furthermore, the TEM findings demonstrated that recombinant ${\alpha}$-gal made no difference in the arrangement of collagen fiber bundles with decellularization. Conclusion: Recombinant B. thetaiotaomicron ${\alpha}$-gal effectively removed ${\alpha}$-gal from bovine pericardium with a small amount under physiological conditions compared to human recombinant ${\alpha}$-gal, which may alleviate the harmful xenoreactive immunologic responses of ${\alpha}$-gal. Recombinant ${\alpha}$-gal treatment had no adverse effects on the mechanical properties of bovine pericardium.

• Journal of Periodontal and Implant Science
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• v.49 no.2
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• pp.90-104
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• 2019

Purpose: The aim of this study was to evaluate clinical and radiographic changes and the survival rate after periodontal surgery using deproteinized bovine bone mineral (DBBM) with 10% collagen or DBBM with a collagen membrane in endo-periodontal lesions. Methods: A total of 52 cases (41 patients) with at least 5 years of follow-up were included in this study. After scaling and root planing with or without endodontic treatment, periodontal regenerative procedures with DBBM with 10% collagen alone or DBBM with a collagen membrane were performed, yielding the DBBM + 10% collagen and DBBM + collagen membrane groups, respectively. Changes in clinical parameters including the plaque index, bleeding on probing, probing pocket depth, gingival recession, relative clinical attachment level, mobility, and radiographic bone gains were evaluated immediately before periodontal surgical procedures and at a 12-month follow-up. Results: At the 12-month follow-up after regenerative procedures, improvements in clinical parameters and radiographic bone gains were observed in both treatment groups. The DBBM + 10% collagen group showed greater probing pocket depth reduction ($4.52{\pm}1.06mm$) than the DBBM + collagen membrane group ($4.04{\pm}0.82mm$). However, there were no significant differences between the groups. Additionally, the radiographic bone gain in the DBBM + 10% collagen group ($5.15{\pm}1.54mm$) was comparable to that of the DBBM + collagen membrane group ($5.35{\pm}1.84mm$). The 5-year survival rate of the teeth with endo-periodontal lesions after periodontal regenerative procedures was 92.31%. Conclusions: This study showed that regenerative procedures using DBBM with 10% collagen alone improved the clinical attachment level and radiographic bone level in endo-periodontal lesions. Successful maintenance of the results after regenerative procedures in endo-periodontal lesions can be obtained by repeated oral hygiene education within strict supportive periodontal treatment.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.34 no.2
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• pp.119-130
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• 2008

Absorbable atelo-collagen sponge $TERUPLUG^{(R)}$, Termo Co. Tokyo, Japan) is inserted in the extraction wound where alveolar bone is exposed. It protects wounds and promotes the formation of granulation. This is made of atelo-collagen, to minimize antigenicity, which is cross-linked by heat treatment for biocompatibility. $TERUPLUG^{(R)}$ consists of between 85 and 95 % of collagen type I and between 5 to 15 % of collagen type III. The raw material for the collagen is derived from bovine skin. It features a sponge block design and is shaped for easy insertion in the extraction wound. This study was designed to find out the bone healing capacity of $TERUPLUG^{(R)}$. We implanted $TERUPLUG^{(R)}$ (experimental group I) and $TERUPLUG^{(R)}$ with rhBMP-2 (experimental group II) in the rabbit cranium defect and then histologically analysed the specimen. The results were as follows. 1. In the 4 weeks, a lot of the newly formed collagen fibers around material of the experimental group I implanted $TERUPLUG^{(R)}$ were observed. But, in the experimental group II implanted $TERUPLUG^{(R)}$ with rhBMP-2, a little of newly formed collagen fibers around material were observed. The cell proliferating activity and apoptosis of the experimental group I, II was positive in and around the implanted material. 2. In the 8 weeks, the amount of newly formed and matured bone in the experimental group II was more observed than the experimental group I and control group. The results of this study indicate that absorbable atelo-collagen sponge ($TERUPLUG^{(R)}$) is relatively favorable bone void filler with biocompatibility and has the better bone healing capacity in case of application with rhBMP-2.

• Microbiology and Biotechnology Letters
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• v.36 no.1
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• pp.34-42
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• 2008

Bovine blood, cell, tissue, and organ are used as raw materials for manufacturing biologics such as biopharmaceuticals, tissue engineered products, and cell therapy. Manufacturing processes for the biologics using bovine materials have the risk of viral contamination. Therefore viral validation is essential in ensuring the safety of the products. Bovine viral diarrhoea virus (BVDV) is the most common bovine pathogen and has widely been known as a contaminant of biologics. In order to establish the validation system for the BVDV safety of biologics, a real-time RT-PCR method was developed for quantitative detection of BVDV contamination in raw materials, manufacturing processes, and final products. Specific primers for amplification of BVDV RNA was selected, and BVDV RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be 1 $TCID_{50}/mL$. The rent-time RT-PCR method was validated to be reproducible and very specific to BVDV. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with BVDV. BVDV RNA could be quantified in CHO cell as well as culture supernatant. Also the real-time RT-PCR assay could detect $10TCID_{50}/mL$ of BVDV artificially contaminated in bovine collagen.

• The Korea Journal of Herbology
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• v.25 no.4
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• pp.129-135
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• 2010

Objectives : The present study was undertaken to determine whether the water extract of Achyranthis Radix, which is the roots of Achyranthes japonica (Achyranthis Radix, AR), is efficacious against collagen-induced arthritis (CIA) in DBA/1J mice. Methods : Mice were immunized with bovine type II collagen and orally treated with AR-W (50 and 100 mg/kg/bw) from days 21 to 42 after immunization. Arthritis was evaluated by arthritic score, histological examination of knee joint and serological markers such as TNF-${\alpha}$, $PGE_2$ and anti-type II collagen (C II)IgG. Results : The results showed that comparing with untreated CIA mice, treated with AR-W significantly suppressed the clinical score and joint tissue pathological damages, reduced the serum levels of TNF-${\alpha}$, $PGE_2$ and anti-C II IgG in CIA-mice. These results suggest that AR-W can effectively alleviate inflammatory response on CIA, and anti-inflammatoy of AR-W can be attributed, at least partially, to the inhibition of inflamamtory mediators, $PGE_2$ and TNF-${\alpha}$, in CIA. Conclusions : This study suggests that AR-W has a therapeutic potential in inflammatory joint diseases such as rheumatoid arthritis.

• The Korea Journal of Herbology
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• v.27 no.3
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• pp.1-6
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• 2012

Objectives : The present study was undertaken to evaluate whether Atractylodis Rhizoma Alba ethanol (ARA-E) extract, which is the pericarp of $Atractylodes$ $japonica$ Koidz. has an effect on collagen-induced arthritis (CIA) in mice. Methods : Male DBA/1J mice were induced by intradermal injection of bovine collagen-II in Freund's incomplete adjuvant (IFA). The CIA mice in the onset of arthritis were treated daily with oral administration of ARA-E extract at dose of 50 mg/kg/bw for 28 days. Arthritis index, histopathological changes and the levels of anti-type II collagen (CII) IgG and inflammatory cytokine, TNF-${\alpha}$ in sera of mice were measured to evaluate the antiarthritic effect of ARA-E. Results : ARA-E extract significantly decreased the arthritic scores and inhibited pathological changes of knee joint tissues in CIA mice. ARA-E extract also significantly decreased the serum levels of anti-CII IgG and TNF-${\alpha}$ in CIA mice. These results indicate that ARA-E extract may effectively prevent arthritic damages in CIA mice, at least partially, by inhibiting the production of autoantibodies and inflammatory cytokine. Conclusions : This studies suggest that ARA-E has a therapeutic potential in inflammatory joint diseases such as rheumatoid arthritis.

• Journal of Pharmacopuncture
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• v.3 no.2
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• pp.27-40
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• 2000

The influence of moxibustion, a traditional Chinese medical treatment, on type II collagen-induced arthritis (CIA) was examined in DBA/1J mice in vivo. Mice were immunized intradermally twice at the 3-week interval with bovine type II collagen (C Il). The main incidence of arthritis started about on day 30 and lasted to day 60 after the first immunization. Moxibustion with three different regimens, was applied at the acupoint equivalent to GV 4 every other day. Moxibustion, from day 0 to day 30 after the first immunization, suppressed the onset and development of arthritis, as well as anti-collagen antibody level. Treatment with moxibustion, from the day 31 to day 60, also resulted in a significant inhibition of progression of arthritis and production of anti-C II antibody. Thirdly we examined the influence of moxibustion on the established arthritis. Moxibustion given from day 61 to day 120, significantly but mildly decreased the anti-C II antibody level in diseased mice, while the bone erosion and joint destruction were not affected. These results indicate that moxibustion could prevent the incidence and attenuates the development of murine CIA.

• Journal of Veterinary Clinics
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• v.27 no.4
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• pp.315-324
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• 2010

The effect of beta-glucan ($Polycan^{TM}$) derived from Aureobasidium pullulans SM-2001 were observed on, collagen-induced rheumatoid arthritis (RA) in DBA mice. Six week-old male DBA/1J mice were immunized by the intradermal injection of $200\;{\mu}g$ of bovine type II collagen with the equal volume of complete Freund's adjuvant at the tail base on day 1. On day 21, the mice were boosted by the intradermal injection of $200\;{\mu}g$ of bovine type II collagen with incomplete Freund's adjuvant. From the first immunization, mice had been administered $Polycan^{TM}$ (21.25, 42.5 and 85 mg/kg), diclofenac and vehicle once a day for 4 weeks, respectively. Collagen-induced hyperimmunities and arthritis signs were markedly and dose-dependently inhibited by treatment of $Polycan^{TM}$ compared with RA control except for tibial cartilages of $Polycan^{TM}$ 21.25 group. $Polycan^{TM}$ effectively inhibited the histopathological changes of collagen-induced arthritis and hyper-immunities.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.863-866
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• 2001

Antioxidative activites and anti skin aging effects of radish(Raphanus sativus L. var. sativus) extracts were studied. The DPPH(1,1-Diphenyl-2-picrylhydrazal) inhibitory effects and CAT(Catalase) activities of hydrophilic radish extracts(DPPH 81.0%, CAT 175units/mg) were significantly higher than lipophilic radish extracts(DPPH 50.0%, CAT 45unitg/mg). But, SOD(superoxide dismutase) activities of hydrophilic radish extracts(SOD 81.3%) were lower than lipophilic radish extracts(SOD 92.3%) In the Bovine Achilles Tendon collagen treated with hydrophilic radish extracts and lipophilic radish extracts, the pyridinoline concentration of collagen were decreased, respectively.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.173-181
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• 2008

Bovine blood, cell, tissue, and organ are used as raw materials for manufacturing biologics such as biopharmaceuticals, tissue-engineered products, and cell therapy. Manufacturing processes for the biologics have the risk of viral contamination. Therefore viral validation is essential in ensuring the safety of the products. Bovine parvovirus (BPV) is one of the common bovine pathogens and has widely been known as a possible contaminant of biologics. In order to establish the validation system for the BPV safety of biologics, a real-time PCR method was developed for quantitative detection of BPV contamination in raw materials, manufacturing processes, and final products. Specific primers for amplification of BPV DNA were selected, and BPV DNA was quantified by use of SYBR Green 1. The sensitivity of the assay was calculated to be $1.3{\times}10^{-1}\;TCID_{50}/mL$. The real-time PCR method was validated to be reproducible and very specific to BPV. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with BPV. BPV DNA could be quantified in CHO cell as well as culture supernatant. Also the real-time PCR assay could detect $1.3{\times}10^0\;TCID_{50}/mL$ of BPV artificially contaminated in bovine collagen. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of BPV contamination during manufacture of biologics.