• Journal of Veterinary Clinics
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• v.24 no.3
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• pp.358-366
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• 2007

This study was performed to investigate the application of bovine amniotic membrane for canine corneal ulcer and to compare bovine amniotic membrane flap to nictitating membrane flap. Corneas were treated with 8.0mm trephine and 100% ethanol was applied and corneal epithelium was removed. This experiment was divided into three groups; control group, nictitating membrane flap group and amniotic membrane + nictitating membrane flap group. To compare corneal healing, fluorescein dye test, histopathological examination and TUNEL assay were performed. The healing size of the corneal epithelium in was largest among groups and it was significantly different with the other groups (P<0.05). Also, inflammatory cell deposits and keratocyte apoptosis in amniotic membrane flap group were smallest, but the width of the corneal epithelium was thickest among groups. Therefore, we tentatively predicted that the amniotic membrane flap would be an alternative effective method for the treatment of superficial canine corneal ulcer.

• Journal of Veterinary Clinics
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• v.23 no.3
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• pp.334-336
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• 2006

An 1.6-year-old female Shih Tzu was presented with corneal perforation, descemetocele, and deep corneal ulcer in the left eye. A bovine amniotic membrane graft which preserved in lyophylized dry form and a third eyelid flap were applied. After 14 days, the cornea was fluorescein-negative. At day 42 postoperatively, only a moderate scar at the corneal center was showed. At the 10-month follow-up, there was clear cornea with a mild scar.

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.645-652
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• 1999

The purpose of the present study was to investigate the effect of bovine amniotic membrane grafts on healing of full-thickness skin wound in dogs. Two $3cm{\times}3cm$ area-matched full-thickness skin wounds were induced bilaterally on the dorsolateral aspect of the trunk of 15 dogs. Chlorhexidine-treated amnion, dried amnion, silver sulfadiazine and 0.9% sterile saline solution were applied on the wound area and examined grossly and histopathologically. Begining 14 days after wounding, amnion applied group had appreciably less amount of inflammatory exudate and hemorrhage than sulfadiazine and saline treated groups. From 14 days after wounding, the degree of wound contraction in amnion groups, especially in the dried amnion group was greater than that of the sulfadiazine and saline treated groups. The percentages of wounds completely healed on 28 days after wounding in saline treated group, chlorhexidine-treated amnion group, dried amnion group and sulfadiazine treated group were 33%, 50%, 83% and 50%, respectively. Microscopically neovascularization and fibrosis were first noticed on 5 days after wounding in the dried amnion group and sulfadiazine treated group, on 7 days in the chlorhexidine-treated amnion group and on 14 days in the saline treated group. Epithelialization in the dried amnion and sulfadiazine treated groups was first noticed on 9 days after wounding, which was faster than that in the other groups. The present study suggests that bovine amniotic membrane, especially dried bovine amnion is effective on healing of full-thickness skin wound in dogs through both wound contraction and epithelialization.

• Archives of Plastic Surgery
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• v.48 no.4
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• pp.448-456
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• 2021

Background Locoregional stem cell delivery is very important for increasing the efficiency of cell therapy. Amnisite BA (Amnisite) is a freeze-dried amniotic membrane harvested from bovine placenta. The objective of this study was to investigate the retention of cells of the stromal vascular fraction (SVF) on Amnisite and to determine the effects of cell-loaded Amnisite in a porcine radiation-induced chronic wound model. Methods Initially, experiments were conducted to find the most suitable hydration and incubation conditions for the attachment of SVF cells extracted from pig fat to Amnisite. Before seeding, SVFs were labeled with PKH67. The SVF cell-loaded Amnisite (group S), Amnisite only (group A), and polyurethane foam (group C) were applied to treat radiation-induced chronic wounds in a porcine model. Biopsy was performed at 10, 14, and 21 days post-operation for histological analysis. Results Retaining the SVF on Amnisite required 30 minutes for hydration and 1 hour for incubation. A PKH67 fluorescence study showed that Amnisite successfully delivered the SVF to the wounds. In histological analysis, group S showed increased re-epithelialization and revascularization with decreased inflammation at 10 days post-operation. Conclusions SVFs had acceptable adherence on hydrated Amnisite, with successful cell delivery to a radiation-induced chronic wound model.

• Archives of Plastic Surgery
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• v.36 no.4
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• pp.385-392
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• 2009

Purpose: In the partial thickness burn management, despite of several advantages, the use of human amniotic membrane has been limited. The authors applied dried bovine amniotic membrane(DBAM) to overcome disadvantages of amniotic membrane for partial thickness burn and compared the effectiveness with cultured allogenic keratinocytes(CAK) that have been recently used for the management of burn. Methods: 16 patients with partial thickness burn, the mean age of 38 ranging 12 to 59 years, between August 2007 and May 2008 were assigned to this study. Either DBAM or CAK was applied, and the secondary dressing was removed on the following day. To compare treatment effect, time for epithelization, Vancouver scar scale and chromameteric results were evaluated. Results: The time for epithelization of DBAM was 10.1 days, that of CAK was 9.1 days, and they were shorter than the previous 2 - 3 weeks. At the follow up Vancouver scar scale was 2.8 for DBAM and 3.0 points for CAK and showed good results. The result of chromameter showed that the $L^*$, $a^*$, and $b^*$ values of the area applied DBAM were 60.1, 13.6, and 13.3, respectively, and the values of the area applied CAK were 60.1, 12.4, and 12.4, respectively. It was found that the skin color of the healed area after burn was darker, the redness was higher, and the yellowness was lower. After dressing, significant side effects were not observed, and in the cases of applying CAK, it was inconvenient as the moving area had to be fixed. Conclusion: With CAK, DBAM has several advantages such as the shortening of the epithelization period, reduction of scar and pigmentation, and convenient application, etc. Thus it is an effective method for the partial thickness burn management.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.346-353
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• 2009

Viral, bacterial, and fungal infection can be transmitted from donor to recipient via transplantation of human amniotic membrane. Therefore human amniotic membrane for transplantation should be disinfected and sterilized before use. The purpose of this study was to examine the efficacy of the disinfection process and sterilization processes used at human tissue bank in the inactivation of viruses, bacteria, and fungi. A variety of experimental model viruses, bacteria, and fungus for human pathogens, including the human immunodeficiency virus type 1 (HIV-1), bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), hepatitis A virus (HAV), porcine parvovirus (PPV), Escherichia coli, Bacillus subtilis, and Candida albicans were all selected for this study. Enveloped viruses such as HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by 70% ethanol treatment, gamma irradiation process, and ethylene oxide (EO) gas sterilization process. Also non-enveloped viruses such as HAV and PPV were effectively inactivated to undetectable levels by gamma irradiation and EO gas treatment. However HAV and PPV showed high resistance to 70% ethanol treatment. E. coli and C. albicans were effectively inactivated to undetectable levels by 70% ethanol treatment, gamma irradiation process, and EO gas treatment. Also B. subtilis was effectively inactivated to undetectable levels by gamma irradiation process and EO gas treatment. However it showed high resistance to 70% ethanol treatment.

• Proceedings of the Korean Society of Veterinary Clinics Conference
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• 2008.10a
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• pp.170-170
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• 2008

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.420-427
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• 2010

A process for manufacturing virally-safe bovine amniotic membrane(BAM) has been developed for biological dressing. BAM was harvested from a healthy bovine placenta, and then the epithelium was removed. The remaining stromal layer was consecutively disinfected with 70% ethanol and 0.05% sodium hypochlorite. The stromal layer was incubated in a decellularization solution containing 0.25%(w/v) trypsin to remove the cellular components. The resulting acelluar BAM was lyophilized to preserve its biochemical and structural integrity. The BAM was packed and exposed to 25 kGy of gamma irradiation for sterilization purpose. Histological, electron microscopical, and biochemical observations showed that the acellualr BAM had intact structural integrity of three dimensional collagen fibers and contained several growth factors, accelerating wound healing, such as EGF (Epidermal growth factor), KGF (Keratinocyte growth factor), and FGF (Fibroblast growth factor). Bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), bovine parainfluenza virus type 3 (BPIV-3), and bovine parvovirus (BPV) were chosen as the biological indicators for validation of viral safety of the acellular BAM. Samples from relevant stages of the production process were spiked with each virus and subjected to viral inactivation processes. Viruses were recovered from the samples and then titrated immediately. All the viruses tested were completely inactivated to undetectable levels within 1 h of 70% ethanol treatment. Enveloped viruses such as BHV, BVDV, and BPIV-3 were more effectively inactivated than BPV by 0.05% sodium hypochlorite treatment. BHV, BVDV, and BPIV-3 were completely inactivated to undetectable levels by 25 kGy of gamma irradiation. Also BPV was effectively inactivated by 25 kGy of gamma irradiation. The cumulative log reduction factors of BHV, BVDV, BPIV-3, and BPV were ${\geq}$13.30, ${\geq}$14.32, ${\geq}$15.22, and ${\geq}$7.57, respectively. These results indicate that the production process for acelluar BAM has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

• Archives of Plastic Surgery
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• v.40 no.1
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• pp.11-18
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• 2013

Background To minimize the inflammatory reaction and improve healing, a new modified dermal substitute composed of an atelocollagen, chondroitin-6-sulfate, and amniotic membrane (AM) was applied to full-thickness skin defects in a pig. Atelocollagen was extracted from bovine skin, and two modified dermal substitutes were generated according to the cross-linking type. Methods The AM-collagen dermal substitutes were characterized and compared with currently used dermal substitutes in a pig skin defect model. There were five experimental groups: dehydrothermal (DHT) cross-linking atelocollagen with the AM on the top (AM-DHT), DHT and chemical cross-linking atelocollagen with the AM on the top (AM-DHT/chemical), Terudermis, Integra, and AlloDerm. After $3{\times}3cm$ full-thickness skin defects on the back of a pig were created, each dermal substitutes dermal substitutes was randomly grafted on the defects. Two weeks after grafting, autologous partial-thickness skin was over-grafted on the neodermis. The take rate of the dermal substitutes, skin, and histological sections were all assessed at 1, 2, and 4 weeks postoperatively. Results More rapid healing and a higher take rate were evident in the AM-DHT and Terudermis groups. Histological examination revealed fewer inflammatory cells and more fibroblast hyperplasia in these two groups. Four weeks after surgery, the amount of newly formed collagen was significantly more appropriate in the AM-DHT group. Conclusions These observations provide supporting evidence that a newly developed amniotic-collagen dermal substitute may inhibit inflammatory reactions and promote wound healing.

• Archives of Plastic Surgery
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• v.49 no.2
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• pp.258-265
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• 2022

Background Chitosan (CS) is a well-known antimicrobial dressing material. Moreover, widely used amniotic membranes contain growth factors beneficial for wound healing. Herein, we created a novel amnion-conjugated CS-alginate membrane dressing and tested its wound healing potency in a diabetic swine model. Methods The bovine amniotic powder growth factor contents were evaluated by protein assay, and the powder's wound healing effects were assessed in vitro by HaCaT cell scratch closure. In vivo, two minipigs developed streptozotocin-induced diabetes. Serial serum glucose measurements and intravenous glucose tolerance tests were performed to confirm their diabetic status. Twelve square-shaped wounds created on each pig's back were randomly divided into control (n = 4), CS (n = 4), and amnion-CS (AC; n = 4) groups and treated accordingly with different dressings. Wound healing in each group was assessed by measuring wound contraction over time, capturing wound perfusion with indocyanine green (ICG) angiography, and histologically analyzing inflammatory markers. Results Amniotic powder elution promoted HaCaT cell migration in the scratch wound model, suggesting its beneficial in vitro wound healing effects. In vivo, the CS and AC groups showed earlier wound contraction initiation and reepithelialization and earlier wound perfusion improvement by ICG angiography than the control group. Additionally, the wound size of the AC group at week 3 was significantly smaller than those in the control group. There was no significant difference in the numbers of acute and chronic inflammatory cells between the groups. Conclusion The amnion-conjugated CS-alginate membrane, as well as CS dressing alone, could be a favorable dressing option for diabetic wounds.