BACKGROUND: Silk cocoon is composed of multiple layers. The natural silk cocoon containing all layers was cut as a rectangular shape as defined as total group. The inner and outermost layers were removed from the total group and the remained mat was defined as the middle group. The objectives of this study was to compare the total group with the middle group as a barrier membrane for the guided bone regeneration. METHODS: The effects of these materials on the cellular proliferation and alkaline phosphatase (ALP) expression of MG63 cells were explored. For comparing bone regeneration ability, bilateral bone defects were created in calvarial areas in ten adult New Zealand white rabbits. The defects were covered with silk membranes of the middle group, with silk membrane of the total group used as the control on the contralateral side. The defects were allowed to heal for 4 and 8 weeks. Micro-computerized tomography (μCT) and histological examination were performed. RESULTS: The middle group exhibited a higher MTT value 48 and 72 h after treatment compared to the total group. ALP expression was also higher in the middle group. The results of µCT and histologic examination showed that new bone formation was significantly higher in the middle group compared to the total group 8 weeks postoperatively (P < 0.05). CONCLUSIONS: In conclusion, the middle layer of the silk cocoon supports guided bone regeneration better than unprocessed silk cocoon.
Adult ; Alkaline Phosphatase ; Bone Regeneration* ; Cell Proliferation ; Humans ; Membranes* ; Osteogenesis ; Rabbits ; Silk*

PURPOSE: This study evaluated powdered burn wound dressing materials from wild silkworm fibroin in an animal model. METHODS: Fifteen rats were used in this experiment. Full-thickness 2x2 cm burn wounds were created on the back of rats under anesthesia. In the two experimental groups, the wounds were treated with two different dressing materials made from silkworm fibroin. In the Control Group, natural healing without any dressing material was set as control. The wound surface area was measured at five days, seven days and 14 days. Wound healing was evaluated by histologic analysis. RESULTS: By gross observation, there were no infections or severe inflammations through 14 days post-injury. The differences among groups were statistically significant at seven days and 14 days, postoperatively (P<0.037 and 0.001, respectively). By post hoc test, the defect size was significantly smaller in experimental Group 1 compared with the Control Group and experimental Group 2 at seven days postoperatively (P=0.022 and 0.029, respectively). The difference between Group 1 and Group 2 was statistically significant at 14 days postoperatively (P<0.001). Group 1 and control also differed significantly (P=0.002). Group 1 showed a smaller residual scar than the Control Group and Group 2 at 14 days post-injury. Histologic analysis showed more re-epithelization in Groups 1 and 2 than in the Control Groups. CONCLUSION: Burn wound healing was accelerated with silk fibroin spun by wild silkworm Antheraea pernyi. There was no atypical inflammation with silk dressing materials. In conclusion, silk dressing materials can be used for treatment of burn wound.
Anesthesia ; Animals ; Bandages* ; Bombyx* ; Burns* ; Cicatrix ; Fibroins* ; Inflammation ; Models, Animal ; Moths* ; Rats* ; Silk* ; Wound Healing ; Wounds and Injuries*

OBJECTIVEAcne vulgaris is a chronic dermatologic problem with multiple factors involved in its pathogenesis. Alternative solutions to acne treatment were instigated by antibiotic resistance despite of its extensive use. Purified bee venom (PBV) has been proposed as a promising candidate for that purpose. The present study was designed to confirm the antibacterial effect of PBV and access the efficacy of cosmetics containing PBV in subjects with acne vulgaris.

METHODSThe skin bacterium Propionibacterium acnes was incubated with PBV at various concentrations and bacterial growth was evaluated using the colony forming unit (CFU) assay. The mechanism of PBV employed in killing P. acnes was examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition, a total of 12 subjects were randomized in a double-blind, controlled trial to receive either cosmetics containing PBV or cosmetics without PBV for two weeks. Evaluations included lesion counts and skin microorganism.

RESULTSPBV exhibited antimicrobial activity in a concentration-dependent manner, reducing the number of P. acnes CFU by approximately 6 logs at a concentration of 0.5 mg. When PBV concentration was higher than 1.0 mg, no P. acnes colonies were spotted on an agar. TEM and SEM of untreated P. acnes illustrated the normal pleomorphic structure, whereas the PBV-treated bacterium lost the integrity of surface architecture. Significant difference (P=0.027) in the grading levels based on numbers of lesion counts for inflammatory and noninflammatory was observed in favour of the PBV group compared with the control group. In terms of average decrement of skin microorganism, subjects receiving cosmetics containing PBV experienced a significant 57.5% decrease of adenosine triphosphate levels, whereas participants receiving cosmetics without PBV experienced a nonsignificant decrease of 4.7%.

CONCLUSIONThese results show that the in vitro actions of antimicrobial activity of PBV were translated in vivo. Cosmetics containing PBV provided a certain degree of efficacy in terms of lesion counts and skin microorganism concentration compared with cosmetics without PBV in subjects with acne vulgaris. PBV may be a good candidate compound for developing therapeutic drug for the treatment of acne vulgaris.

Acne Vulgaris ; drug therapy ; microbiology ; Adolescent ; Adult ; Anti-Infective Agents ; therapeutic use ; Bee Venoms ; therapeutic use ; Child ; Cosmetics ; Double-Blind Method ; Humans ; Propionibacterium acnes ; drug effects

Acne vulgaris is a chronic dermatologic problem with multiple factors involved in its pathogenesis. Alternative solutions to acne treatment were instigated by antibiotic resistance despite of its extensive use. Purified bee venom (PBV) has been proposed as a promising candidate for that purpose. The present study was designed to confirm the antibacterial effect of PBV and access the efficacy of cosmetics containing PBV in subjects with acne vulgaris.

Oral administration of royal jelly (RJ) promotes wound healing in diabetic mice. Concerns have arisen regarding the efficacy of RJ on the wound healing process of normal skin cells. In this study, a wound was created by scratching normal human dermal fibroblasts, one of the major cells involved in the wound healing process. The area was promptly treated with RJ at varying concentrations of 0.1, 1.0, or 5 mg/ml for up to 48 hrs and migration was analyzed by evaluating closure of the wound margins. Furthermore, altered levels of lipids, which were recently reported to participate in the wound healing process, were analyzed by HPTLC and HPLC. Migration of fibroblasts peaked at 24 hrs after wounding. RJ treatment significantly accelerated the migration of fibroblasts in a dose-dependent manner at 8 hrs. Although RJ also accelerated the migration of fibroblasts at both 20 hrs and 24 hrs after wounding, the efficacy was less potent than at 8 hrs. Among various lipid classes within fibroblasts, the level of cholesterol was significantly decreased at 8 hrs following administration of both 0.1 ug/ml and 5 mg/ml RJ. Despite a dose-dependent increase in sphinganines, the levels of sphingosines, ceramides, and glucosylceramides were not altered with any concentration of RJ. We demonstrated that RJ enhances the migration of fibroblasts and alters the levels of various lipids involved in the wound healing process.
Administration, Oral ; Animals ; Ceramides ; Cholesterol ; Chromatography, High Pressure Liquid ; Fatty Acids ; Fibroblasts ; Glucosylceramides ; Humans ; Mice ; Skin ; Sphingosine ; Wound Healing

Injection inoculation protocols for fruit body formation of Cordyceps militaris (C. militaris) were investigated to improve the incidence of infection in the silkworm species Bombyx mori (B. mori). Injection, with suspensions of C. militaris hyphal bodies into living silkworm pupae, was used to test for fruit body production. Use of Daeseungjam rather than Baegokjam or Keumokjam varieties of B. mori is thought to be suitable for infection by C. militaris. From mounting, nine-day-old to 11-day-old pupae showed the best incidence of infection with a 100 microL injection volume. Silkworm pupae injected with a hyphal suspension concentration of more than 2 x 105 colony-forming unit (cfu) recorded a greater than 96% incidence of infection. Also, fruit bodies of C. militaris were induced and produced at a light intensity between 500 and 1,000 lx.
Bombyx ; Cordyceps ; Fruit ; Incidence ; Light ; Pupa ; Stem Cells ; Suspensions

Recombinant human epidermal growth factor (rhEGF) stimulates the proliferation and migration of epithelial cells in human cell culture systems and animal models of partial-thickness skin wounds. This study investigated the effect of a topical rhEGF ointment on the rate of wound healing and skin re-epithelialization in a rat full thickness wound model, and verified whether or not the rhEGF treatment affected both myofibroblast proliferation and collagen synthesis in the dermis. When rhEGF (10 microgram/g ointment) was applied topically twice a day for 14 days, there was significantly enhanced wound closure from the 5th to the 12th day compared with the control (ointment base treatment) group. A histological examination at the postoperative 7th day revealed that the rhEGF treatment increased the number of proliferating nuclear antigen immunoreactive cells in the epidermis layer. In addition, the immunoreactive area of alpha-smooth muscle actin and the expression of prolyl 4-hydroxylase were significantly higher than those of the control group. Overall, a topical treatment of rhEGF ointment promotes wound healing by increasing the rate of epidermal proliferation and accelerating the level of wound contraction related to myofibroblast proliferation and collagen deposition.
Actins/genetics/metabolism ; Administration, Topical ; Animals ; Cell Proliferation/drug effects ; Collagen/*biosynthesis ; Epidermal Growth Factor/*administration&dosage/*pharmacology ; Gene Expression Regulation ; Male ; Myoblasts, Skeletal/*drug effects ; Proliferating Cell Nuclear Antigen/genetics/metabolism ; Rats ; Rats, Sprague-Dawley ; Wound Healing/*drug effects

Free amino acids in epidermis function as a major component of Natural Moisturizing Factor (NMF), which maintains the optimal level of water in skin even at the low humidity. In fact, the depletion of free amino acids is reported in the epidermis of atopic dermatitis, the skin condition involving dryness. As an effort searching the dietary source for improving the level of water and free amino acid in epidermis, the dietary effects of silk protein, sericin (S) and fibroin (F) on trans epidermal water loss (TEWL), and plasma and epidermal levels of free amino acids were compared in this study. Thirty of male NC/Nga mice, an animal model of atopic dermatitis, were divided into three groups: group CA as an atopic control with control diet, group S: 1% sericin diet and group F: 1% fibroin diet. Ten of male BALB/c mice were served as group C (control group) with control diet. All mice were fed on diet and water ad libitum for 10weeks. Dry skin condition was established in group CA as TEWL was increased (148.7% of group C). In parallel, epidermal level of glutamate, one of major amino acids functioning as NMF, was dramatically decreased and epidermal levels of methionine and alanine were inversely elevated. Dietary supplementation of sericin (group S) reduced TEWL at the similar level with group C and increased epidermal levels of glutamate as well as serine and glycine, the other major amino acids as NMF. Despite a marked decrease of methionine and alanine, the reduction of TEWL and epidermal levels of glutamate, serine and glycine of group F were less than of group S. Furthermore, in contrast to similar levels of other free amino acids in plasma and epidermis of group S and group C, plasma and epidermal levels of other free amino acids, specifically phenylalanine, isoleucine, cysteine and tyrosine in epidermis of group F, were significantly higher than of group C. Together, our data demonstrate that dietary supplementation of sericin is more effective at improving dry skin condition that paralleled with the normalization of free amino acids in plasma and epidermis of NC/Nga mice.
Alanine ; Amino Acids ; Animals ; Cysteine ; Dermatitis, Atopic ; Diet ; Dietary Supplements ; Epidermis ; Fibroins* ; Glutamic Acid ; Glycine ; Humans ; Humidity ; Isoleucine ; Male ; Methionine ; Mice* ; Models, Animal ; Phenylalanine ; Plasma* ; Sericins* ; Serine ; Silk* ; Skin ; Tyrosine

In this present study, we show that 3HK-induced reactive oxygen species (ROS) accumulation and caspase activation lead to apoptotic cell death. Pretreatment with N-acetylcysteine (NAC), an effective antioxidant, significantly attenuated 3HK-induced apoptosis by way of a reduction of ROS accumulation and caspase activity. SKN-SN cells were protected from 3HK-induced cytotoxicity by heat shock protein (HSP). HSP90 effectively attenuated 3HK-mediated ROS accumulation and apoptosis. In addition, the protective effect of HSP90 was abolished by pretreatment with HSP90 anti-sense oligonucleotide, but not when pretreated with anti-senses for other HSPs. These results suggest that HSP90 protects SKN-SH cells from 3HK-induced cytotoxicity by reducing ROS levels and caspase activity.
Acetylcysteine ; Apoptosis ; Cell Death* ; Heat-Shock Proteins* ; Hot Temperature* ; Reactive Oxygen Species*

Endogenously or exogenously generated oxidative stress impair organs, especially brain. Also, the oxidative stress appears to be a negative factor on normal brain function, like memory and cognition. Our result showed that BF-7, extracted from Bombyx mori, effectively diminished oxidative stress, leading to the protection of neuron from reactive oxygen species donated by FeSO4 . Clinical experiments showed that BF-7 significantly improved memory and cognitive functions of normal adults in a dose-dependent manner. Thus, our results suggest that BF-7 play a role in the improvement of brain functions by removing oxidative stress, and provide therapeutic potential role of BF-7 to protect nervous system from oxidative damage.
Adult ; Apoptosis ; Bombyx ; Brain ; Cognition ; Humans ; Memory* ; Nervous System ; Neurons* ; Oxidative Stress* ; Reactive Oxygen Species