혈장 중 Cefprozil의 이성체인 cis-Cefprozil과 trans-Cefprozil의 동시분석법

조애리,문희경,이성창 德成女子大學校 藥學硏究所 2004 藥學論文誌 Vol.15 No.1

Cefprozil is a broad-spectrum oral beta-lactam cephalosporin consisting of cis-- and trans-isomers in and approximately 90:10 ratio. To establish an assay condition for a Bioequvalence study of cefprozil, a column liquid chromatographic (HPLC) method using UV detection for the determination of the cis- and trans-isomers of cefprozil in blood has been developed. The HPLC apparatus used in this study was Waters 2695 Separation Module (Alliance) system. Samples were analyzed with dual absorbance detector (280㎚). C8 (150×4.6㎜, Luna 5㎛, Phenomenex) column with column inlet filter (3㎜×O.5㎛), provided a good resolution, The mobile phase was composed of acetonitrile : glacial acetic acid : water: (5:2:93 as v/v/v, pH 2.7). The flow rate was 1.0㎖/min. Typical retention times with Luna column were 10.8, 14.8 and 20.5min for cis-cefprozil, trans-cefprozil and cepharolidine (internal standard), respectively. For the data manipulation, Waters Millennium program was employed. Calibration curves for the determination of cis-cefprozil and trans isomers of cefprozil in plasma showed a good linearity at a concentration range from 0.05 to 15㎍/㎖ (r²=0.999) and 0.01-1.5㎍/㎖ (r²=0.998). The lower detection limits for cis- and trans-cefprozil were 0.05 and 0.01㎍/㎖, respectively.

설파디아진은의 피부세포 증식 및 화상모델에 있어서의 상처치유과정에 미치는 영향

조애리 德成女子大學校 藥學硏究所 2002 藥學論文誌 Vol.13 No.1

Cyto-toxic effect of silver sulfadiazine (Ag-SD) on kerationcytes and its implication of wound healing process were investigated in 2^nd degree burn hairless mouse model. As a dermal model, HaCat (immortalized Keratinocytes) monolayer culture in DMEM with 10% FBS was used. Cyto-toxicity of Ag-SD was estimated by measuring the cell viability using neutral red assay after adding the drug. The 2^nd degree burn was prepared on hairless mouse back skin (1 cm diameter)and dressings with Ag-SD were applied for 96 hr. The process of re-epithelialization and the presence of inflammatory cells were investigated and histology with Hematoxylin-Eosin staining was performed. Ag-SD displayed highly cyto-toxic effect on cultured HaCat cells in a concentration dependent manner (1-100 ㎍/mL). Topical applicantion of Ag-SD(2%) could control the infection: no inflammatory cells were obsevered in histology. However the cyto-toxic effect of Ag-SD on skin cells induced the impairment in epidermal regeneration.

아세메타신 및 인도메타신의 혈장 중 동시분석법

조애리,문희경 德成女子大學校 藥學硏究所 2003 藥學論文誌 Vol.14 No.1

Acemetacin is a new potent non-steroidal anti-inflammatory compound which is used for the treatment of arthritis and rheumatic diseases. To establish an assay condition for a Bioequivalence study of acemetacin, a column liquid chromatographic (HPLC) method using UV detection for the determination of acemetacin and its metabolites indometacin in blood has been developed. The HPLC apparatus used in this study was a Waters 2695 Separation Module (Alliance) system. Samples were analyzed with Dual-λ-Absorbance Detector (UV254 nm). C_(18) (250×4.6mm, Luna 5μm, Phenomenex) column which maintained at 40℃, provided a good resolution. The mobile phase was composed of 0.02M 인산용액 (pH 4.5):MeOH=45:55. The flow rate was 1.4ml/min. Typical retention times with Luna column were 23, 31 and 34 min for flubiprofen (internal standard), acemetacin and indometacin, respectively. For the data manipulation, Waters Millennium program was employed. Calibration curves for the determination of acemetacin and indometacin in plasma showed a good linearity at a concentration range from 100ng/ml to 4000ng/ml(r²=0.999). The lower detection limits for both compounds were 100ng/ml.

Enhancing Dermal Matrix Regeneration and Biomechanical Properties of 2nd Degree-Burn Wounds by EGF-Impregnated Collagen Sponge Dressing

Ae-Ri Cho Lee 대한약학회 2005 Archives of Pharmacal Research Vol.28 No.11

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 μg/1.7 cm2. 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, 2nd degree- burn of diameter 1cm 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 ± 35.6 vs. 411.5 ± 57.0 Fmax, gmm-2) and skin resilience (11.3 ± 1.4 vs. 6.5 ± 0.6 mJ/mm2) 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, 2nd degree-burns enhanced the dermal matrix formation.

Gravimetric Analysis and Differential Scanning Calorimetric Studies on Glycerin-induced Skin Hydration

Ae-Ri Cho Lee,Hee Kyung Moon 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.11

A thermal gravimetric analysis (TGA) and a differential scanning calorimetry (DSC) were carried out to characterize the water property and an alteration of lipid phase transition of stratum corneum (SC) by glycerin. In addition, the relationship between steady state skin permeation rate and skin hydration in various concentrations of glycerin was investigated. Water vapor absorption-desorption was studied in the hairless mouse stratum corneum. Dry SC samples were exposed to different conc. of glycerin (0-50%) followed by exposure to dry air and the change in weight property was monitored over time by use of TGA. In DSC study, significant decrease in ∆H of the lipid transition in 10% glycerin and water treated sample: the heat of lipid transition of normal, water, 10% glycerin treated SC were 6.058, 4.412 and 4.316 mJ/mg, respectively. In 10% glycerin treated SCs, the Tc of water shifts around 129oC, corresponding to the weakly bound secondary water. In 40% glycerin treated SC, the Tc of water shifts to 144oC corresponding to strongly bound primary water. There was a good correlation between the hydration property of the skin and the steady state skin flux with the correlation coefficient (r2=0.94). As the hydration increased, the steady state flux increased. As glycerin concentration increased, hydration property decreased. High diffusivity induced by the hydration effect of glycerin and water could be the major contributing factor for the enhanced skin permeation of nicotinic acid (NA).

Phospholipid Polymer, 2-Methacryloyloxyethyl Phosphorylcholine and Its Skin Barrier Function

Ae-Ri Cho Lee 대한약학회 2004 Archives of Pharmacal Research Vol.27 No.11

The effect of poly[2-methacryloyloxyethyl phosphorylcholine] (pMPC) on the skin permeation property was investigated by performing in vitro skin permeation study of a model drug, nicotinic acid (NA). Effect of pMPC polymer in donor solution on skin permeation rates was evaluated using side-by-side diffusion cells. Also, the structural alterations in the stratum corneum (SC), inter-lamellar bilayer (ILB) and dermis layers in pMPC-treated and -untreated skin sections were investigated with transmission electron microscopy (TEM). The permeation profile of NA without pMPC in donor solution showed biphasic mode: initial 1st phase and 2nd hydration phase. The sudden, more than 10-fold increase in flux from the initial steady state (43.5 μg/cm2/hr) to the 2nd hydration phase (457.3 μg/cm2/h) suggests the disruption of skin barrier function due to extensive hydration. The permeation profile of NA with 3% pMPC in the donor solution showed monophasic pattern: the steady state flux (10.9 μg/cm2/h) without abrupt increase of the flux. The degree of NA permeation rate decreased in a concentration-dependent manner of pMPC. TEM of skin equilibrated with water or 2% pMPC for 12 h showed that corneocytes are still cohesive and epidermis is tightly bound to dermis in 2% pMPC-treated skin, while wider separation between corneocytes and focal dilations in inter-cellular spaces were observed in water-treated skin. This result suggests that pMPC could protect the barrier property of the stratum corneum by preventing the disruption of ILB structure caused by extensive skin hydration during skin permeation study.

Size matters: differential property of hyaluronan and its fragments in the skin- relation to pharmacokinetics, immune activity and wound healing

Cho Lee Ae-Ri 한국약제학회 2023 Journal of Pharmaceutical Investigation Vol.53 No.3

Background Numerous adverse immune reactions after hyaluronan (HA) applications have been reported, highlighting the need for investigating the differences in metabolism and pharmacokinetics between homeostasis and tissue injury and inflammation conditions. Understanding the mechanisms underlying the pro- or anti-inflammatory activity of HA depending on its fragment size and the association between HA fragment size and its pharmacokinetics and therapeutic properties is critical to its use without adverse immune reactions. Area covered This paper discusses the differential synthesis and degradation of HA depending on physiological conditions and HA fragment size and the strategies to minimizing adverse immune reactions. Given HA is an important skin component, its skin penetration, dermal pharmacokinetics and differential effects of native HA and fragments on wound healing and tissue regeneration are discussed. Expert opinion In ideal homeostatic conditions, the proportion of high-molecular-weight HA (HMW HA; ~106 Da, 1.24 M Da) is high, and it rarely binds with immune cells. However, tissue injury and inflammation lead to the generation of small HA fragments (~ 100 kDa and 10 kDa), which exhibit inflammatory, carcinogenic, and tissue fibrotic properties. As HA is an indispensable extracellular matrix (ECM) component of skin, specific skin absorption enhancement induced by conversion of the α-helix to a ß-sheet structure of HA and hydration of stratum corneum weakens the SC barrier. Given different cell types react differently to cellular stresses. Currently, no general guideline to prognosticating serious adverse reactions after HA application exists. Therefore, we believe that normalization of HA metabolism and an in vitro model evaluating the specific inflammatory cytokine profile and gene expression level in macrophage cell lines of tested tissue after HA exposure would help in devising a clinical strategy to minimize the serious adverse reactions after HA application.

Controlled Partial Skin Thickness Burns: Rabbit Ear as a 2^nd Degree Burn Wound Model for Studies of Topical Therapy

Lee, Ae-Ri Cho,Moon, Hee-Kyung 한국약제학회 2006 Journal of Pharmaceutical Investigation Vol.36 No.5

Controlled Partial Skin Thickness Burns: Rabbit Ear as a 2^nd Degree Burn Wound Model for Studies of Topical Therapy

Cho, Lee Ae-Ri,Moon, Hee-Kyung The Korean Society of Pharmaceutical Sciences and 2006 Journal of Pharmaceutical Investigation Vol.36 No.5

This study was designed to prepare an animal model for partial thickness bum wound which can be employed for testing topical therapy. We first evaluated whether rabbit ear and mouse back skin wound model could differentiate the wound healing process in terms of degree of re epithelialization, required days for complete wound closure, presence of scarring. $2^{nd}$ degree wet bum were prepared on mouse back skin and rabbit ear by applying 5 mL hot water($85{\pm}0.1^{\circ}C$) for 7 see followed by 5 mL ice-cold 0.5% acrynol solution for cooling and disinfecting the inflicted area. After removing the dead epidermis layer at 24 hr, tested dressings were applied for specified time and wound progression was investigated. In mouse model, wound contraction was the primary wound closing mechanism, which is quite different from human wound healing process. In rabbit ear model, epidermal regeneration was the major wound healing process rather than wound contraction and the difference in wound healing property among tested dressings could be clearly demonstrated. A rabbit ear model could differentiate the wound progression among open, occluded and epidermal growth factor(EGF) treated wound. Four sites of circular wound(diameter: 1 cm) on the anterior part of rabbit ear could be employed for the comparative wound healing study. For obtaining reproducible bum wound, degree of bum depth and bum sites should be carefully controlled in addition, employing rabbits of same strain and weight. The result suggests that rabbit ear could be employed as a reliable and human-resembled wound model.

Enhancing Dermal Matrix Regeneration and Biomechanical Properties of $2^{nd}$ Degree-Burn Wounds by EGF-Impregnated Collagen Sponge Dressing

Cho Lee Ae-Ri The Pharmaceutical Society of Korea 2005 Archives of Pharmacal Research Vol.28 No.11

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.