In order to formulate an aqueous topical preparation of epidermal growth factor(EGF) for the treatment of open wound and bum, the stability of EGF in aqueous vehicles containing various stabilizers was evaluated and the pharmacological activity of gel preparations formulated with poloxamer 407 was determined with wound model. Various additives, which are known as potent stabilizers for proteins and polypeptides so far, were used to increase the stability of EGF in aqueous vehicles. The contents of EGF in the vehicles containing stabilizers were determined with an HPLC method after the storage at $37^{\circ}C$. EGF was more stable in ultrapure water than RO water or saline. All the additives studied resulted in deleterious effects on EGF stability. Therefore, it was speculated that any additives or impurities in the vehicle made EGF unstable. However, nitrogen purge of solution increased the stability of EGF in aqueous vehicles. The aqueous topical preparations of EGF were formulated with poloxamer 407 as a gel base in saline. Gelatin or amastatin was employed as a protease inhibitor. The pharmacological effect of EGF gel was studied with open wound model in mice. EGF preparations, made of oleaginous base or poloxamer gel base, showed significant healing effect compared to the control group(p<0.05). The addition of protease inhibitor in poloxamer 407 gel resulted in significant healing effect compared to the gel without it(p<0.05). Body weights of mice treated with EGF preparation were increased at the first day after the formation of open wound, while those of the control group were decreased. The EGF gel made of poloxamer 407 containing a pretense inhibitor would be a promising aqueous topical preparation for EGF.
Journal of Dental Rehabilitation and Applied Science
/
v.20
no.1
/
pp.31-41
/
2004
The osseointegration in implant therapy is achieved following general wound healing mechanism. Platelet play a major role in wound healing process. In addition to blood clot formation, they secrete many growth factors which regulate the attachment, proliferation and differentiation of nearly all cell types. The use of these growth factors is now known to be very effective methods to improve the cellular activity. Platelet-rich plasma which is made with the newly developed technique concentrating platelets 3-folds or more is also proven to be very effective method to stimulate and accelerate the healing of bone and soft tissue. Previous study proved that platelet-rich plasma enhanced the cellular attachment by inducing fibronectin, vitronectin from osteoblast. So, this study was aimed to investigate the effect of platelet-rich plasma on the cellular proliferation and differentiation in vitro. The effect on the proliferation was evaluated by MTT assay. To evaluate autocrine and paracrine effect, conditioned medium was made and compared. By measuring alkaline phosphatase activity, the effect on the cellular differentiation was evaluated. The results were as following: The cellular proliferation of osteoblast cell line increased depending on the concentration of platelet-rich plasma and conditioned medium. The alkaline phosphatase activity increased depending on the concentration of platelet-rich plasma and conditioned medium. These findings imply that platelet-rich plasma enhance the cellular proliferation and differentiation and maximize the cellular activity by using the autocrine and paracrine effect.
Journal of the Korean Society of Food Science and Nutrition
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v.43
no.3
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pp.374-380
/
2014
In this study, the anti-inflammatory effects of Lespedeza cuneata extract on macrophages and wound-healing in wound-induced animal experiments were investigated. In an anti-inflammatory test, 0.1 mg/mL of Lespedeza cuneata extract did not affect growth of RAW 264.7 cells, and Lespedeza cuneata extract suppressed nitric oxide (NO) generation from inflammation-induced macrophages in a concentration-dependent manner. Wounds on the skin of rats were treated with vehicle containing Lespedeza cuneata extract (SSP), vehicle (SCO), and commercial ointment (CCO). The wound and scar sizes in the SSP group were significantly reduced in comparison to the SCO and CCO groups (P<0.05). The epidermis and dermis of the SSP group also recovered faster than the SCO group based on Masson's trichrome staining. The gene expression levels of vascular endothelial growth factor (VEGF) decreased and transforming growth factor-beta 1 (TGF-${\beta}1$) increased in wound tissue from the SSP group compared to that from the SCO group. These results show that Lespedeza cuneata extract accelerates wound-healing through anti-inflammatory activity and induction of collagen regeneration as well as reduces the scar area surrounding wounds. Accordingly, Lespedeza cuneata extract could be useful as a cosmeceutical in the cosmetic industry.
Adequate acidic environment in wound healing prevents the inflammation of virus, increases the cell activity, promotes cell proliferation and regular rearrangement of fibroblast, and results in matured epithelialization. In this study, we prepared dressing materials consisting of pectin and carboxymethylcellulose (CMC) with varied ratios. These dressing materials showed different pH values according to the composition ratio. The effect of acidity of pectin/CMC dressing materials on wound healing rates, degree of epithelialization, collagen deposition, and so on, in 3 types of wound models (fresh surgical wounds, $3^{rd}$ degree burn wounds, and infection wounds) were investigated by animal tests. From the results of wound contraction, wound healing, and epithelialization, it can be deduced that dressing material having pectin/CMC ratio of 16/19 (pH 4.67) is most effective among the 3 types of wound models.
Antarctic krill has a strong proteolytic enzyme system, which comes from a combination of several proteases. This powerful activity can be easily detected by krill's superior post mortem autolysis. Mammalian skin consists of epidermis and dermal connective tissue, and functions as a barrier against threatening environments. A clot in a wound site of the skin should be removed for successful skin regeneration. Epithelial cells secrete proteases to dissolve the clot. In previous studies Antarctic krill proteases were purified and characterized. The proteolytic enzymes from Antarctic krill showed higher activity than mammalian enzymes. It has been suggested that these krill clean up the necrotic skin wound to induce a natural healing ability. The enzymes exhibited additional possibilities for several other biomedical applications, including dental plaque controlling agent and healing agent for corneal alkali burn. Considering that these versatile activities come from a mixture of several enzymes, discovering other proteolytic enzymes could be another feasible way to enhance the activity if they can be used together with krill enzymes. Molecular cloning of the krill proteases should be carried out to study and develop the applications. This review introduces possible roles of the unique Antarctic krill proteases, with basic information and suggestion for the development of an application to skin regeneration.
Go, Young Bin;Lee, Ji Hwan;Oh, Han Jin;Kim, Yong Ju;An, Jae Woo;Chang, Se Yeon;Song, Dong Cheol;Cho, Hyun Ah;Cho, Jin Ho
Korean Journal of Agricultural Science
/
v.48
no.3
/
pp.567-574
/
2021
In this study, we evaluated the wound healing rate and, inflammatory cells effects of by Abeliophyllum distichum Nakai (ADN) extract in mice. We also assessed the stability of the ADN extract upon exposure to sunlight. Treatments were as follows: 1) CON (only saline solution), T1 (CON + 0.0125% ADN extract), T2 (CON + 0.05% ADN extract), and T3 (CON + 0.5% ADN extract). A 4 mm punch was used in the central part of the dorsal area to separate it from the subcutaneous tissue, causing a full-thickness skin wound. An amount of 1 mL of each sample was sprayed onto the treatment section of the wound with a pipette every day from the day of wound creation, with proper application ensured using brush. In the stability test, the pH was measured at 1, 4, and 8 weeks after exposing the samples of each treatment section to sunlight considering, the higher concentrations of the ADN extract. The results of this study indicate that the effectiveness of the wound contraction rate in the mice to which the ADN extract was applied was low. Moreover, the stability of the sample containing a high concentration of the ADN extract could not be verified. In addition, no significant results were obtained in the inflammatory reaction assessment. Therefore, additional research focusing on wound contraction, stability, and inflammatory cell outcomes of the ADN extract is needed.
The purpose of this study was to investigate the effect of the high voltage pulsed Current (HVPC) stimulation on the healing rate and the proliferative activity of keratinocytes and IGF-I mRNA expression of an incisional wound in rat skin. Twenty male Sprague-Dawley rats ($265{\sim}290g$) were randomly divided into HVPC (n=10) and control group (n=10). Rats received 10 mm length of full-thickness incision wound on the back under the anesthesia. The HVPC group received electrical stimulation with a Current intensity of 50 V at 100 pps for a duration of 30 minutes, while the control group was given the same treatment without electricity for a week. Polarity was negative in first three days and positive thereafter. The wound length was measured and evaluated as percentage. The mean number of nucleolar organizer regions (NORs) per nucleus and level of IGF-I mRNA expression were calculated. The mean percent of wound closure were $51.17{\pm}17.76%$ and $80.71{\pm}11.91%$, respectively, in the sham treated control and HVPC stimulated groups (t=-4.308, P<0.001). The mean NOR number per nucleus of the keratinocytes in the control and HVPC group were $1.85{\pm}0.20$ and $2.70{\pm}0.23$, respectively (t=8.638, P<0.001). The IGF-I mRNA level were $0.76{\pm}0.44$ and $1.32{\pm}0.41$, respectively, in the control and HVPC stimulated wounds (t=2.906, P<0.01). There was a positive correlation between the mean NOR number per nucleus and IGF-l mRNA level with a Pearson product moment correlation coefficient of 0.72 (P<0.05). These findings suggest that the HVPC may activate the rRNA of the basal keratinocytes and upregulate the IGF-I mRNA levels by alteration of the electrical environment, and it may increase proliferative activity of the keratinocytes in the skin wound of the rat.
The purpose of this study w8s to evaluate the effects of pulsed electromagnetic energy(Diapulse) and microcurrent on the wound healing in rabbits. 15 domestic rabbits were randomly assigned to the PRME(n=5). MC(n=5) and CON(n=91 group. Each rabbits were anesthetized with lidocaine HCL $2\%$. Skin wounds were created laterally on the back of IS domestic rabbits(33cm). From 24 hours after being injured, the rabbits of the PEME group were irradiated with an intensity of 3 at a 300 pulses per second, which were applied for 15 minutes every day during the 12 days. The MC group were stimulated with an intensity of $50{\mu}A$ at frequency of40 pulses per second, which were applied for 15 minutes every day during the 12 days. The CON group were not stimulated. The rabbits were sacrificed and the incised wound pans were processed appropriately for the light microscopic examination on the 3rd day, 6th day and 12th day before the beginning of wound treatment. The areas of wound were measured with metric graph paper. The results were as tallows. 1 The PRME and MC group compared with control group showed that wound closure rate increased on 6th, 12th day. 2. It was found that the CON group did not show a complete maturation and had a chronic inflammatory response. Judging from the irregularity of intercellular space and the loose alignment of connective tissue. these findings showed that wound healing was delayed. 3. It showed that inflammatory cells. fibroblasts and epithelial cells activity rapidly processed in the PEME group compared with the CON group. It was found that the PEMI; group showed a advanced remodeling of epithelial layer and a positive repair of connective tissue. 4. It showed that fibroblasts, epithelial cells and inflammatory cells activity rapidly processed in the MC group compared with the CON group. It was found that the MC group showed a improved remodeling of epithelial layer and a dense connective tissue.
Background Reactive oxygen species cause serious damage to the physiological function of tissues. Determination of total antioxidant capacity of skin tissue is one of the determinants of damaged tissue function. Mast cells (MCs) are one of the groups of cells that are invited to the site of injury. The healing process begins with the rapid release of various types of MCs' intermediate factors at the site of injury. Bone marrow mesenchymal stem cell (BMMSC) production and secretion have been shown to regenerate the skin. The aim of this research was to evaluate the wound-healing and antioxidant effects of BMMSCs per MCs. Methods Fifty-four albino Wistar male rats were divided into three groups: (1) nonsurgery, (2) surgery, and (3) surgery + BMMSCs. Groups 2 and 3 were operated with a 3 × 8 cm flap and in group 3, cell injections (7 × 109 cell injection at the time of surgery) were performed. After days 4, 7, and 15, percentage of the surviving tissue, histological characteristics, superoxide dismutase (SOD) activity, and amount of malondialdehyde (MDA) were measured in the groups. For results, Graph Pad Prism 8 software was used, and data were analyzed and compared by analysis of variance and Tukey test. Results BMMSCs' application decreased the amount of MDA, increased SOD activity and survival rate of the flaps, and improved the histological characteristics. Conclusion This study revealed the protective effects BMMSCs alongside MCs against oxidative stress on the survival of the flaps. However, for clinical use, more research is needed to determine its benefits.
The effects of agitation rate and agitation time on the synthesis of sphingosome were studied. As increase in the agitation rates 4,000, 6,000, 8,000, 10,000 and 12,000 rpm the viscosity of sphingosome were decreased. The most sufficient agitation rate was 8,000rpm for which micell viscosity and stability. The effect of agitation time on the sphingosome viscosity and stability was investigated by changing the agitation times 2, 4, 6, 8 and 10 minute at 8,000rpm. 4 minute was the most sufficient for the viscosity and stability. The sphingolipid activity of cutaneous wound healing in impaired mice was examined. As a result, we could prove that phytosphingosine-HCl medically worked on wound healing well. For the phytosphingosine-HCl, it was found that the experimentally determined medical action more effective than that of tetra-acetylphytosphingosine.
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