Skin Of Male Wistar Rats Research Articles

Administration of Arabika Coffee Leaf Extract (Coffea Arabica) Reduces Tumor Necrosis Factor Alpha (TNF-α) and Sunburn Cell Levels in Male Wistar Rats (Rattus norvegicus) Exposed to Ultraviolet-B Rays

Background: Ultraviolet (UV) radiation has acute and chronic impacts on the skin. However, UVB exposure can be prevented with available natural ingredients, especially those containing antioxidants. One of the natural ingredients that contains active compounds as antioxidants is arabica coffee leaves, which contain flavonoids that have antioxidant effects. Methods: This study was an experimental research with a post-test-only control group design, using 30 male Wistar rats (Rattus norvegicus) which were divided into 5 groups; the control group (P0), treatment group 1 (P1) were given placebo, while group 2 (P2), group 3 (P3) and group 4 (P4) was given UVB rays and given ethanol extract cream from Arabica coffee leaves at a dose of 2,5%, 5%, and 10% respectively. Results: The average TNF- α levels in group P0, P1, P2, P3, and P4 were 97,88±1,19 ng/mL, 162,41±1,87 ng/mL, 138,04±1,03 g/mL, 124,71±2,65 ng/mL and 107,18±1,52 ng/mL. The average sunburn cells in P0, P1, P2, P3, and P4 were 4,0±0,13, 4,07±0,27, 2,60±0,25, 1,67±0,15 and 1,33±0,12 per field view. The administration of 10% Arabica coffee leaf extract cream was the most effective in reducing TNF-α levels by 34.01% (p<0.001) and has the lowest sunburn cells per field view (1,33±0,12 cell/field view; p < 0,001). Conclusion: Administration of ethanol extract cream from Arabica coffee leaves (Coffea arabica) can reduce TNF-α levels and the number of sunburn cells in the skin of male Wistar rats (Rattus norvegicus) exposed to UVB radiation.

Characterization and biocompatibility evaluation of acellular rat skin scaffolds for skin tissue engineering applications.

Utilization of acellular scaffolds, extracellular matrix (ECM) without cell content, is growing in tissue engineering, due to their high biocompatibility, bioactivity ad mechanical support. Hence, the purpose of this research was to study the characteristics and biocompatibility of decellularized rat skin scaffolds using the osmotic shock method. First, the skin of male Wistar rats was harvested and cut into 1 × 1 cm2 pieces. Then, some of the harvested parts were subjected to the decellularization process by applying osmotic shock. Comparison of control and scaffold samples was conducted in order to assure cell elimination and ECM conservation by means of histological evaluations, quantification of biochemical factors, measurement of DNA amount, and photographing the ultrastructure of the samples by scanning electron microscopy (SEM). In order to evaluate stem cell viability and adhesion to the scaffold, adipose-derived mesenchymal stem cells (AD-MSCs) were seeded on the acellular scaffolds. Subsequently, MTT test and SEM imaging of the scaffolds containing cultured cells were applied. The findings indicated that in the decellularized scaffolds prepared by osmotic shock method, not only the cell content was removed, but also the ECM components and its ultrastructure were preserved. Also, the 99% viability and adhesion of AD-MSCs cultured on the scaffolds indicate the biocompatibility of the decellularized skin scaffold. In conclusion, decellularized rat skin scaffolds are biocompatible and appropriate scaffolds for future investigations of tissue engineering applications.

A shea butter-based ketamine ointment: The antidepressant effects of transdermal ketamine in rats

The delayed onset of monoaminergic antidepressants and disadvantages of traditional administration routes created the need for alternative non-invasive delivery methods with rapid onset therapeutic effect. Ketamine attracted attention as a fast-acting glutamatergic antidepressant with ideal physiochemical properties for alternative routes of administration. However, there is no sufficient data for its transdermal use in depression. In this proof-of-concept study, we investigated the antidepressant effects of transdermal ketamine delivered via a novel ointment with skin protective, emulsifying and permeation enhancing properties. A shea butter-based 5% (w/w) ketamine ointment or a drug-free vehicle ointment were applied to the shaved dorsal skin of male Wistar rats for 2 days, twice a day. Behavioral despair, locomotor activity and anxiety-like behavior were respectively assessed in the forced swim test (FST), open field test (OFT), and elevated plus maze (EPM). The pharmacokinetic profile of the ointment was analyzed with high-performance liquid chromatography. Transdermal ketamine ameliorated behavioral despair without altering general locomotor activity and anxiety-like behavior, showing that skin-friendly drug carriers like shea butter may constitute promising alternatives to current routes of delivery for ketamine. Tested transdermal method aims to provide more sustainable drug delivery for long-term treatment schedules. Future studies can investigate its long-term use, side effects and abuse liability.

Peppermint Leaf Extract Cream Increased Transforming Growth Factor β (TGF-β) Expression and Collagen Amount In The Male Wistar Rat's Skin Exposed to UVB

Peppermint leaves contain flavonoids, menthofuran, and other phenolic compounds that have antioxidant activity and are believed to inhibit collagen degradation in the skin. This study investigated the efficacy of a 5% peppermint leaf extract cream in increasing the transforming growth factor β (TGF-β) expression and collagen amount as indicators of collagen maintenance in male Wistar rats' skin exposed to UVB. Method A randomized post-test-only control group design was performed on male Wistar rats, local strains, aged 2-3 months, with body weights ranging from 180 to 200 grams. According to the sample calculation, 21 rats were randomly divided into three groups. The first group is the control group (without any treatment). The second group was treated with UVB without cream (UVB-only group), and the third group was treated with UVB + 5% peppermint leaf extract cream on exposed skin. After four weeks of treatment, the tissue samples were examined. The expression of TGF-β was evaluated by immunohistochemical examination, and the amount of collagen was examined by Picro-Sirius red staining. Comparative tests were performed on the measurement results of the three groups. The results showed a significant difference in the mean TGF-β expression and collagen amount among the three groups (p<0.001). The group exposed to UVB + 5% peppermint leaf extract cream had the highest mean TGF-β expression and collagen amount (77.41±5.79% and 82.57±2.18%, respectively), followed by the control group (63.17±12.22% and 74.05±7.15%) and the UVB-only group (50.54±11.10% and 65.65±5.56%). Administration of 5% peppermint leaf extract cream effectively increased TGF-β expression and collagen amount in the skin of male Wistar rats exposed to UVB compared to the control and the UVB-only groups.

Administration of Sugar Apple Leaf Extract Cream (Annona squamosa L.) Increased Superoxide Dismutase (SOD) Activity and Decreased Skin Matrix Metalloproteinase-1 (MMP-1) Activity in Male White Rats (Rattus norvegicus) Wistar Strain Exposed to Ultraviolet B Light

Background: Ultraviolet B rays from sunlight causes increased levels of matrix metalloproteinase-1 (MMP-1). Superoxide dismutase (SOD) also will decrease to counteract these free radicals. Sugar apple (Annona squamosa L.) leaf extract showed significant antioxidant and anti-collagenase activity in vitro. Methods: This is a posttest only control group design study. Thirty male white Wistar rats (Rattus norvegicus), 12 weeks old, 150-200 grams were divided into five groups, 1 group without UVB exposure and without cream, and 4 groups was exposed to UVB 3 times a week and by giving cream 2 times a day for 2 weeks, 20 minutes before exposure and 4 hours after exposure. Rats were fixated in a box. The UVB dose was 65 mJ/cm2 for 65 seconds, with total 390 mJ/cm2. Skin samples were tested using ELISA method. Results: Negative control (P1) compared to Sugar apple (Annona squamosa L.) leaf extract cream 1%. (P2), 3% (P3), and 5% (P4) concentration for SOD levels were 2.3015 ± 0.32501 vs. 2.7817 ± 0.26899 vs. 4.7752 ± 0.65218 vs. 6.0890 ± 0.63946, respectively and for MMP-1 levels were 3.0753 ± 0.51113 vs. 2.3037 ± 0.15556 vs. 1.5840 ± 0.25473 vs. 0.9198 ± 0.16604, p < 0.001), respectively. Conclusion: Sugar apple (Annona squamosa L.) leaf extract cream 3% and 5% can increase SOD levels and 1%, 3%, and 5% can reduce MMP-1 levels in the skin of male Wistar rats (Rattus norvegicus) exposed to UVB light. Concentration 3% is the most effective. Keywords: sugar apple leaf extract, SOD, MMP-1, ultraviolet B rays

Eucalyptus globulus essential oil increased neovascularization and vascular endothelial growth factor expression in male wistar rats exposed to UVB

Background: Eucalyptus globulus essential oil contained 1,8-cineol, which may accelerate wound healing. This study aimed to prove Eucalyptus globulus essential oil's effect on increasing neovascularization and Vascular Endothelial Growth Factor (VEGF) expression in Wistar rats skin exposed to ultraviolet B (UVB).
 Methods: A Post-test only controls group study design was applied on 36 male Wistar rats, aged 3-4 months old, weighing 150-200 grams. The animal subject was exposed to UVB (65 mJ/cm2, 65 seconds for seven days), injured on their back using a punch biopsy with a diameter of 10 mm, and divided into two groups. For seven days, on the wound, the control group was given 2 cc of 0.9% NaCl twice a day, while the treatment group was given Eucalyptus globulus essential oil (2 cc). Both groups were given oral Amoxicillin three times a day. The skin tissue was taken on day eight to assess neovascularization and VEGF expression.
 Results: The results show that neovascularization in the treatment group was higher than in the control group (p < 0.05). Meanwhile, the median VEGF expression in the treatment group was also higher than in the control group (p < 0.05).
 Conclusions: Topical administration of Eucalyptus globulus essential oil increased neovascularization and VEGF expression in the skin of male Wistar rats injured and exposed to UVB. Further research is needed to assess the effectiveness of the essential oil in enhancing neovascularization and VEGF expression in humans with multiple exposures at different doses to optimize the wound-healing effect.

MORPHOFUNCTIONAL REBUILDING OF FIBROUS STRUCTURES OF RAT'S SKIN DERMIS UNDER 3D-SCAFFOLD IMPLANTATION BASED ON POLYPROLACTONE

The use of various scaffolds makes it possible to model the future fibrous framework of the newly formed regenerate, and also serves as a substrate for the colonization of the cellular component. The development of tissue engineering in regenerative medicine requires an understanding of more precise mechanisms of the formation of the connective frame at the site of the defect. The aim of this study was to study the morphofunctional rearrangement of the fibrous structures of the dermis of the skin of rats, in response to for the implantation of a 3D-scaffold based on polyprolactone. The study was carried out on the skin of male Wistar rats at different times after implantation of a 3D-scaffold based on polyprolactone. The biomaterial with the implantable scaffold was studied using scanning electron microscopy. As a result of the study, it was revealed that active collagenogenesis is determined around the structures of the 3D-scaffold based on polyprolactone. By the 14th day, a large number of circularly directed collagen fibers are visualized, which are the basis for further building a connective tissue capsule around the fibers. Conclusions are drawn about the effectiveness of using a 3D-scaffold based on polyprolactone. This implant can also be effective for the colonization of allogeneic cultured fibroblasts and the creation of biomedical tissue engineering products on its basis.

Cyclooxygenase-2 Modulates Glycosaminoglycan Production in the Skin During Salt Overload.

Sodium (Na+) can accumulate in the skin tissue, sequestered by negatively charged glycosaminoglycans (GAGs). During dietary salt overload, the amount and charge density of dermal GAG molecules – e.g., hyaluronic acid (HA) and chondroitin sulfate (CS) – increases; however, the regulation of the process is unknown. Previously, it has been demonstrated that the level of cyclooxygenase-2 (COX-2) activity and the content of prostaglandin E2 (PGE2) are elevated in the skin due to high-salt consumption. A link between the COX-2/PGE2 system and GAG synthesis was also suggested. We hypothesized that in dermal fibroblasts (DFs) high-sodium concentration activates the COX-2/PGE2 pathway and also that PGE2 increases the production of HA. Our further aim was to demonstrate that the elevation of the GAG content is ceased by COX-2 inhibition in a salt overloaded animal model. For this, we investigated the messenger RNA (mRNA) expression of COX-2 and HA synthase 2 enzymes as well as the PGE2 and HA production of DFs by real-time reverse transcription PCR (qRT-PCR) and ELISA, respectively. The results showed that both high-sodium concentration and PGE2 treatment increases HA content of the media. Sodium excess activates the COX-2/PGE2 pathway in DFs, and COX-2 inhibition decreases the synthesis of HA. In the animal experiment, the HA- and CS disaccharide content in the skin of male Wistar rats was measured using high performance liquid chromatography-mass spectrometry (HPLC-MS). In the skin of rats receiving high-salt diet, the content of both HA‐ and monosulfated-CS disaccharides increased, whereas COX-2 inhibition blocked this overproduction. In conclusion, high-salt environment could induce GAG production of DFs in a COX-2/PGE2-dependent manner. Moreover, the COX-2 inhibition resulted in a decreased skin GAG content of the salt overloaded rats. These data revealed a new DF-mediated regulation of GAG synthesis in the skin during salt overload.

DYNAMICS OF RAT SKIN FORMATION DURING THE TRANSITION FROM PRENATAL TO EARLY POSTNATAL ONTOGENESIS

The peculiarities of the skin formation as an organ are the basic knowledge for studying the aging and regeneration process. During studying this issue it is necessary to take into account the exogenous factors affecting the skin, which, in turn, is a "barrier organ". The study was carried out on the skin of male Wistar rats in the late prenatal and early postnatal periods. At the initial stages of its formation, the skin undergoes its morphofunctional changes such as thickening of the epidermis and the appearance of the stratum corneum, as well as thickening of the dermis and changes in the collagen fibers shape that make up its structure. The formation of the skin as a full-fledged "barrier organ" occurs in early postnatal ontogenesis.

Protective Effect of Lemon (Citrus limon L.) Ethanol Extract Cream as an Antioxidant Against Exposure to Ultraviolet B Rays in the Skin of Male Wistar (Rattus norvegicus) Rats

Background: Indonesia is an archipelagic country and one of the tropical countries that get sun exposure all the time. Sun exposure can have beneficial and detrimental effects in life. Exposure to radiation can cause acute effects in the form of erythema through the inflammatory process. Antioxidants are substances that can protect the body from damage caused by ROS. Natural antioxidants can be found in vegetables and fruits, one of which comes from lemon extract (Citrus limon L.). Lemon extract is known to have various chemical contents with active compounds in the form of flavonoids and phenols which can act as antioxidants. The aim of this research was to determine the protective effect that can be produced by lemons on UVB exposure in the skin of male wistar rats.Methods: This research uses the true experimental post test only control group design method. Samples were divided into three treatment concentrations, namely ethanol extract of lemon 5%, 10%, and 20%.Result: After testing for normality, the significance value was obtained (p <0.05). Based on the results of the normality and homogeneity test, the results of the data distribution are not normal and the homogeneous tests of the hypotheses used are the Kruskal-Wallis non-parametric test. Kruskal-Wallis non-parametric test results showed a significant difference with the significance value (p = 0.001). The best degree of erythema score was found in the cream of 10% ethanol extract of lemon with an average of 0.8 ± 0.84.Conclusion : The ethanol extract of lemon (Citrus limon L.) cream in a certain dosage has a significant effect on reducing the erythema degree score in the back skin of male Wistar rats (Rattus norvegicus) after exposure to UVB rays.

Java plum (Syzygium cumini) extract cream effect on decreasing the number of sunburn cell on the dorsal skin of male Wistar rat (Rattus norvegicus) exposed to UV-radiation

Background: The primary acute damage due to UV radiation is keratinocyte apoptosis or sunburn cell. Previous studies showed that antioxidants inhibit UV damage. Java plum (Syzygium cumini) which is found in Indonesia, was known to have a high concentration of antioxidants. This study was aimed to assess the Java plum ethanol extract cream effects on the number of sunburn cells on the dorsal skin of male Wistar rats that exposed to UV-B.
 Methods This experimental research was a post-test only control group design that was conducted on 30 male rats. Groups were divided into control, placebo, 5%, 10%, and 20% ethanol extract cream concentrations (P1, P2, P3, P4, and P5, respectively). The number of sunburn cell were then observed and recorded based on visual evaluation of the skin biopsy under a light microscope.
 Results: The mean results of P1, P2, P3, P4, and P5 were 132.67, 98.67, 118.67, 104.00, and 54.67 respectively. The control, placebo, 5%, and 10% ethanol extract cream showed no significant result (p>0.05). In contrast, 20% ethanol extract cream showed a protective effect (p<0.05).
 Conclusion: Java plum ethanol extract cream with 20% dose could decrease the number of sunburn cells in the skin.

Evaluation of post-surgical healing in rats using a topical preparation based on extract of Musa sapientum L., Musaceae, epicarp

Considering that oral preparations made with peel green bananas (e.g. flour and extracts) demonstrated healing effects on mucous membranes and skin, this study evaluated the healing and the antimicrobial property of a topical preparation based on extract of Musa sapientum L., Musaceae, (apple banana) in surgically induced wounds in the skin of male Wistar rats, 100 g. The extract was obtained by decoction, the presence of tannins was detected by phytochemical screening and 10% of the extract was incorporated into the carbopol gel (CMS gel). The processes of healing and bacterial isolation were evaluated in the following experimental groups: control (no treatment), treatment with placebo or with the CMS gel. The healing of surgical wounds treated with the CMS gel was faster when compared with the control and placebo groups and the treatment with CMS gel also inhibited the growth of pyogenic bacteria and enterobacteria in the wounds. The results indicate that the extract of Musa sapientum epicarp has healing and antimicrobial properties (in vivo), probably, due to tannins.

Effects of diode laser therapy on the acellular dermal matrix

Acellular dermal matrix (ADM) was subcutaneously implanted into calvarian skin of male Wistar rats (n = 40). Low-level laser (lambda 685 nm, 4 J/cm(2)) was locally applied in experimental group (n = 20) above the skin flap. Grafts were harvested at 1, 3, 7 and 14 days after surgery and underwent histological analyses. In treated animals, the extent of edema and the number of inflammatory cells were reduced (P < 0.05). The amount of collagen in graft treated with low-level laser were significantly higher than those of controls (P < 0.05) and were statistically more prominent on the 14th day after surgery. The mean count of fibroblasts was significantly higher in the low-laser therapy group within the 3rd day, showing a marked influx of fibroblasts into area. In conclusion, wound healing of the ADM appear to be positively affected by laser therapy.