• Archives of Pharmacal Research
• /
• v.26 no.10
• /
• pp.855-860
• /
• 2003

The effect of silver sulfadiazine (SSD) on the proliferation of human dermal fibroblast (HDF) was studied to determine the impact of the drug on the wound healing process and dermal mechanical strength. Human dermal fibroblasts were cultured to 80% confluency using DMEM with 10% FBS and viability of the cell was estimated using neutral red assay. In addition, the $2^{nd}$ degree burn wound was prepared on the anterior part of rabbit ear skin and dressings containing SSD were applied for 96 h. Presence of inflammatory cells and degree of re-epithelialization were investigated in the wound. After 15 day of the induction of burn wounds, the treated area was excised and dermal mechanical strength was quantitatively measured with a constant speed tensiometer. SSD was found to be highly cyto-toxic in cultured HDF cells. The topical application of SSD (2%) could control the infection as evidenced by the lack of accumulation of inflammatory cells in histological evaluation. Therefore, these observations suggested that the impairment of dermal regeneration and decreased mechanical strength of dermal tissue was resulted from the cyto-toxic effect of SSD on dermal cells. Since the decreased mechanical strength may lead to reduction in resilience, toughness and maximum extension of the tissue, the identification of optimum dose for SSD that limits infection while minimizes the cyto-toxic effect may be clinically relevant.

• Journal of Veterinary Clinics
• /
• v.30 no.6
• /
• pp.403-408
• /
• 2013

The objective of the present study is to detect the effect of ${\beta}$-glucan originated from Aureobasidium on the proliferation and collagen production in human dermal fibroblast cells with wound repopulation in vitro. The proliferative effects were assessed using a MTT assay as well as cell counts at 24 and 48 hr after treatment. Hydroxyproline was measured as an index of procollagen production with reverse-phase high pressure liquid chromatography. Oncostatin M was used as a reference agent. In glucagon treated group, dose-dependent and significant increase of optical density or fibroblast cell numbers was demonstrated, when compared with those of control from 0.1 mg/ml concentration. In addition, the numbers of cells which had migrated into the wound defects were more significantly and dose-dependently increased than those of non-treated control. However, no meaningful effects on the procollagen production were observed.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.26 no.1
• /
• pp.59-80
• /
• 2000

본 연구는 dermal equivalent를 이용하여 AHA류(lactic acid, glycolic acid, citric acid 및 malic acid) 및 보습 원료(sodium hyaluronate, glycerine, natural carbohydrate complex)에 대한 wound healing 효과 및 그 기작을 알아보기 위한 것이다. AHA류 및 보습 원료의 독성은 dermal equivalent를 이용하여 m assay를 실시한 후 적정 농도를 petri dish위에 조성한 dermal equivalent에 처리하였다. 그리고 wound healing 효과를 알아보기 위하여 collagen 수축률을 측정하였다. 또한 collagen 합성을 촉진하는 것으로 잘알려진 vitamin C도 함께 처리하였다. AHA류의 경우 평균 pH 2-3으로 매우 낮기 때문에 4N NaOH를 이용하여 pH를 6-7사이로 조절한 sample를 함께 실험하였다. 보습 원료의 경우 전반적으로 10% 이상의 높은 NR$_{50}$보여 주었다. 특히 sodium hyaluronate(1% stock solution)의 경우에는 16% NR$_{50}$를 보여 주었다. pH를 6-7로 조절한 AHA류의 경우에는 전반적으로 0.6% 내외의 NR$_{50}$를 보이는 가운데 lactic acid는 상대적으로 높은2% NR$_{50}$를 보여 주었다. Collagen 수축률 측정 실험 결과에서는 2% sodium hyaluronate(1% stock solution)가 대조군에 비하여 처리 후 2일째 25%내외의 향상된 수축률을 보여 주었다. pH를6-7로 조절한 AHA류 중 0.1%의 malic acid의 경우에서는 대조군에 비하여 처리 후 1일째 및 2일째 각각 28% 및 35%의 수축률을 보여 주었으며 pH를 6-7로 조절한 0.1% vitamin C에서도 유사한 결과를 보여 주었다. 반면에 pH를 6-7로 조절한 0.1% citric acid의 경우에는 10-20%의 낮은 수축률을 보여 주었다. MTT assay를 이용한 UV 조사 후 pH를 6-7로 조절한 AHA류의 repairing UV damage 효과에 대한 실험에서 0.1% 및 0.01%의 malic acid와 0.01% citric acid은 irradiation control에 비하여 약 10% 이상 세포수를 증가시켰다. 그러나 예외적으로 citric acid의 경우 0.1% 농도에서 오히려 20%내외로 세포수가 감소되는 경향을 보여주었다. 그리고 lactic acid 및 glycolic acid는 두드러지는 효과를 나타내지 않았다. Collagen 합성을 측정 실험에서는 pH를 6-7로 조절한 AHA류에서는 대조군에 비하여 상대적으로 12-19% 더 합성을 촉진하였다. 반면에 pH가 2-3인 AHA류의 경우에는 대조 군과 유사하거나 조금 낮은 합성율을 보여 주었다.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.23 no.1
• /
• pp.30-41
• /
• 2010

Purpose: This study was to observe the effect of "herbal decoction for sitz bath" on dermoepidermal recovery to wound tissue in rat's skin. Methods: The samples were assigned to 3 groups: control group : without any treatment, positive control group : potarose 10% solution, experiment group : herbal decoction for sitz bath. We made the open wound of $2{\times}2cm^2$ size that cut deep into the dermis. Treating the open wound for 17 days, we observed the size of the wound diminishing. On 17th days, the cell viability was measured by MTT assay. The effect anti-inflammatory and dermoepidermal recovery were examined by H&E staining, immunohystochemical staining for MIP-2, FGF. Results: The experiment group showed more recovery from the open wound comparing the control group and the positive control group on 10th days after wounding. But there was not remarkable difference between the experiment and positive control group after 17th days post-wounding. The number of MIP-2 positive reacted cell were significantly decreased and that of FGF positive reacted cell were significantly increased than positive control group at 17th days. Conclusion: According to these results, we finally concluded that "herbal decoction for sitz bath" could be effective in recovery to wound tissue.

• Journal of Veterinary Clinics
• /
• v.18 no.4
• /
• pp.418-423
• /
• 2001

To investigate the modulation of nerve growth factor (NGF) during corneal epithelial wound healing and the effect of topical NGF on corneal epithelial wound healing in dogs. An axial epithelial defect was created in the right eye using 6mm axial corneal mechanical debridement while the left served as an unwounded control. The tears were collected from both eyes during 1 week and the corneal epithelium was processed for the measurement of NGF at day 0 and 7. The NGF content of tears and corneal epithelium was determined by enzyme-linked immunosorbent assay. In another experiment, the animals were divided into 3 groups. The right eyes in each group were treated every six hours with 200 ug/ml of recombinant human (rh) NGF, murine NGF, or 600 ug/ml of anti-NGF blocking antibody. The left eye of each animal was treated with bovine serum albumin (BSA) to serve as controls. Wound healing was analyzed using NIH image software. Tear NGF was markedly increased in the wounded eyes, relative to tears from control eyes during the early healing period. The NGF content of the corneal epithelium was elevated in the wounded eye (p=0.024). Time to wound closure and rate of epithelial migration were not significantly different between the NGF treated or the NGF antibody treated, and the control BSA treated eyes. Corneal epithelial wounding increased NGF content only on the wounded side during the early healing period. Neither topical recombinant human or murine NGF affected corneal epithelial wound healing in the normal dog.

• Proceedings of the PSK Conference
• /
• 2002.10a
• /
• pp.327.3-328
• /
• 2002

Transforming growth factor-${\beta}$ (TGF-${\beta}$), a hormonally active polypeptide found in normal and transformed tissues. regulates cellular growth and phenotyphic plasticity. We have previously shown that H-ras. but not N-ras. induces invasive phenotype in MCF10A human breast epithelial cells. In this study. we wished to examine the effect of TGF-${\beta}$ on H-ras-induced invasion and motility in MCFI 10A cells by performing in vitro invasion assay and wound migration assay. (omitted)

• The Korean Journal of Pesticide Science
• /
• v.9 no.4
• /
• pp.429-436
• /
• 2005

As Sclerotinia sclerotiorum causing cucumber sclerotinia rot was the fastest in the mycelial growth at $25^{\circ}C$, its pathogenicity was strong at the same temperature among several temperatures. All the isolates of Sclerotinia sclerotiorum showed a strong pathogenicity against cucumber fruits, which was confirmed by a disk assay and a wound assay. A wound assay was superior to a disk assay to develop the assay system for assessing the fungicidal activity of several fungicides against Sclerotinia sclerotiorum. In a disk assay, it was very difficult to assess the fungicidal activity, because the pathogenicity of isolates used in the experiment was very strong. At 500 and $3.0{\mu}g/mL$, the activity of dichloflouanid and the mixture of carbendazim and diethofencarb against cucumber sclerotinia rot was 14.3 and 42.3%, respectively, by using a disk assay. However, at same concentration two fungicides showed the high controlling activity as 100 and 92.5%, through a wound assay in a laboratory. Also, the activity of two fungicides was good against cucumber sclerotinia rot in the greenhouse where cucumber plants were cultivated in the field, showing the control value as 91.1 and 82.9% at 100 and $825{\mu}g/mL$, respectively. All the isolates of Sclerotinia sclerotiorum from cucumber fruits sampled in the polyvinyl house were subjected to monitoring for the resistance to 7 fungicides. The $EC_{50}$ value of 7 fungicides was as follows: fenhexamid; $0.13{\mu}g/mL$, procymidon and iprodione; 0.18 and $0.24{\mu}g/mL$, carbendazim and the mixture of carbendazim and diethofencarb; 0.13과 $0.05{\mu}g/mL$, iminoctadine and dichlofluanid; 1.94 and $8.95{\mu}g/mL$. Ultimately it was not found that resistant isolates of Sclerotinia sclerotiorum were appeared in the field.

• Archives of Pharmacal Research
• /
• v.28 no.11
• /
• pp.1311-1316
• /
• 2005

To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGF­treated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.

• YAKHAK HOEJI
• /
• v.37 no.5
• /
• pp.532-537
• /
• 1993

Angiogenesis refers to the formation of new capillary blood vessels, or neovascularization occurring under various physical conditions, such as development of the embryo, formation of corpus luteum, wound healing and pathological conditions including tumor growth and metastases, hemangiomas, diabetic retinopathy, rheumatoid arthritis. There are many evidences that angiogenesis is important for the progressive growth of solid tumors and also permits the shedding of metastatic tumors from the primary site. Thus, treatment of angiogenesis inhibitors might be a novel strategy for tumor growth inhibition. Normal vascular endothelial cells are in a state of differentiation and angiogenic endothelial cells migrate and proliferate, and they subsequently differentiate into vessel-forming quiescent phenotype cells, Therfore, it was speculated that a modifier of cell differentiation could also affect angiogenesis. In order to identify new antiangiogenic factors, the research was conducted to estimate the inhibitory activities of cell differentiation agents by means of chick embryo chorioallantoic membrane(CAM) assay. Hence, we have established the CAM assay for the screening of antiangiogenic agents. Using the CAM assay, we found that ursolic acid, a tumor cell differentiation-inducing agent, showed a markedly inhibitory effect on chick embryonic angiogenesis.

• Korean Journal of Pharmacognosy
• /
• v.46 no.1
• /
• pp.59-64
• /
• 2015

Wound healing is a complex process that includes inflammation, granulation tissue formation, re-epithelialization, and remodeling. We reported previously that BuOH fraction from Hydnocarpi Semen (HS) crude extract exhibited wound healing activity in animal ulcer models. In this study, we investigated whether BuOH fraction activates keratinocyte and fibroblast via wound closure test and migration assay. In the scratch test, BuOH fraction accelerated the closure of a monolayer wound scratch at $100{\mu}g/mL$. After treatment with BuOH fraction for 18 h, keratinocytes showed a increase in migration at $25{\mu}g/mL$, whereas the migration of fibroblast increased significantly at $100{\mu}g/mL$ of BuOH fraction compared to control. The mechanism that the BuOH fraction of HS helps to promote healing by inflammation is possibly associated with the migration of keratinocyte and fibroblast.