Search results for: acellular dermal matric

Authors: Tim Bolle, Franziska Kreimendahl, Thomas Gries, Stefan Jockenhoevel

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The therapeutic treatment of extensive, deep wounds is limited. Autologous split-skin grafts are used as a so-called ‘gold standard’. Most common deficits are the defects at the donor site, the risk of scarring as well as the limited availability and quality of the autologous grafts. The aim of this project is a tissue engineered dermal-epidermal skin replacement to overcome the limitations of the gold standard. A key requirement for the development of such a three-dimensional implant is the formation of a functional capillary-like network inside the implant to ensure a sufficient nutrient and gas supply. Tailored three-dimensional warp knitted spacer fabrics are used to reinforce the mechanically week fibrin gel-based scaffold and further to create a directed in vitro pre-vascularization along the parallel-oriented pile yarns within a co-culture. In this study various three-dimensional warp knitted spacer fabrics were developed in a factorial design to analyze the influence of the machine parameters such as the stitch density and the pattern of the fabric on the scaffold performance and further to determine suitable parameters for a successful fibrin gel-incorporation and a physiological performance of the scaffold. The fabrics were manufactured on a Karl Mayer double-bar raschel machine DR 16 EEC/EAC. A fine machine gauge of E30 was used to ensure a high pile yarn density for sufficient nutrient, gas and waste exchange. In order to ensure a high mechanical stability of the graft, the fabrics were made of biocompatible PVDF yarns. Key parameters such as the pore size, porosity and stress/strain behavior were investigated under standardized, controlled climate conditions. The influence of the input parameters on the mechanical and morphological properties as well as the ability of fibrin gel incorporation into the spacer fabric was analyzed. Subsequently, the pile yarns of the spacer fabrics were colonized with Human Umbilical Vein Endothelial Cells (HUVEC) to analyze the ability of the fabric to further function as a guiding structure for a directed vascularization. The cells were stained with DAPI and investigated using fluorescence microscopy. The analysis revealed that the stitch density and the binding pattern have a strong influence on both the mechanical and morphological properties of the fabric. As expected, the incorporation of the fibrin gel was significantly improved with higher pore sizes and porosities, whereas the mechanical strength decreases. Furthermore, the colonization trials revealed a high cell distribution and density on the pile yarns of the spacer fabrics. For a tailored reinforcing structure, the minimum porosity and pore size needs to be evaluated which still ensures a complete incorporation of the reinforcing structure into the fibrin gel matrix. That will enable a mechanically stable dermal graft with a dense vascular network for a sufficient nutrient and oxygen supply of the cells. The results are promising for subsequent research in the field of reinforcing mechanically weak biological scaffolds and develop functional three-dimensional scaffolds with an oriented pre-vascularization.

Keywords: fibrin-gel, skin replacement, spacer fabric, pre-vascularization

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Authors: Yan Li, Qi-Fan Jia, Jie Zhou

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In today’s China, primary and secondary teachers are living a hard life with high pressure but low payment, which results in a sense of unfair and less satisfaction of life. However, teachers’ life satisfaction is a significant factor of their mental health and plays an important role in the development and progress of the society. This study was aimed to explore the effect of teachers’ perception of social justice on life satisfaction. 450 primary and secondary teachers from China were measured with life satisfaction scales, social justice scales, income satisfaction scale, job satisfaction scale, pressure condition scale, and major life event scale. Results showed their pressure is significantly higher than average, while life satisfaction, job satisfaction, income satisfaction and perceived social justice are lower. Hierarchical regression analysis showed that demographic variables, i.e., gender, age, education level and matric status, and factors related to occupation, i.e., professional title, school type and working hours per day cannot predict teachers’ life satisfaction. Teachers who had worked for 11-20 years had a lower life satisfaction compared to those with 1-5 years working experience. However, social status, monthly household income, income satisfaction, as well as job satisfaction were positively related to life satisfaction, whereas pressure condition was negatively related to it. After controlling for demographic factors and individual attitudes, social justice still had a positive effect on life satisfaction, among which distributive justice played a more important role than procedural justice. The suggestions on teachers’ condition in China and the implications for education reform to improve teachers’ mental health are discussed.

Keywords: life satisfaction, mental health, primary and secondary teachers, social justice

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Authors: W. M. Buba, J. G. Nangbes, J. P. Butven

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This research was undertaken to evolve community based awareness on the arsenic contamination from agricultural practices in Communities of Bokkos local government area. Contaminated farmland soil samples were collected from the surface for tailings and at various depths (50, 100, 150 cm intervals) in eight holes drilled in each farm at different locations using hand auger. A total of sixty- four (64) soil samples were collected from eight (8) different communities. A standard titrimetric method was applied for the determination of arsenic. It was found that the average concentration of arsenic in the surface soil (0-150cm) for the entire study areas was 0.0525mg/kg with range 0.0425 -0.0601mg/kg which is well above the recommended the soil to plant concentration guideline range of 2.3 – 4.3 x10-4 mg/kg value. This indicates that the arsenic concentration in the study areas does pose health risk for agricultural practices via potential bioaccumulation in plant food crops. However, some risks measures could follow the arsenic occurrence through direct exposure such as those resulting from the inhalation, oral or dermal intake of arsenic during agricultural practices and in the course of stay on the contaminated soil.

Keywords: agrochemicals, arsenic, bokkos, contamination, soil

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Authors: Dmitry Yu. Korulkin, Raissa A. Muzychkina

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Bioactive substances of plant origin can be one of the advanced means of solution to the issue of combined therapy to inflammation. The main advantages of medical plants are softness and width of their therapeutic effect on an organism, the absence of side effects and complications even if the used continuously, high tolerability by patients. Moreover, medial plants are often the only and (or) cost-effective sources of natural biologically active substances and medicines. Along with other biologically active groups of chemical compounds, essential oils with wide range of pharmacological effects became very ingrained in medical practice. Essential oil was obtained by the method hydrodistillation air-dry aerial part of Polygonum L. plants using Clevenger apparatus. Qualitative composition of essential oils was analyzed by chromatography-mass-spectrometry method using Agilent 6890N apparatus. The qualitative analysis is based on the comparison of retention time and full mass-spectra with respective data on components of reference oils and pure compounds, if there were any, and with the data of libraries of mass-spectra Wiley 7th edition and NIST 02. The main components of essential oil are for: Polygonum amphibium L. - γ-terpinene, borneol, piperitol, 1,8-cyneole, α-pinene, linalool, terpinolene and sabinene; Polygonum minus Huds. Fl. Angl. – linalool, terpinolene, camphene, borneol, 1,8-cyneole, α-pinene, 4-terpineol and 1-octen-3-ol; Polygonum alpinum All. – camphene, sabinene, 1-octen-3-ol, 4-carene, p- and o-cymol, γ-terpinene, borneol, -terpineol; Polygonum persicaria L. - α-pinene, sabinene, -terpinene, 4-carene, 1,8-cyneole, borneol, 4-terpineol. Antibacterial activity was researched relating to strains of gram-positive bacteria Staphylococcus aureus, Bacillus subtilis, Streptococcus agalacticae, relating to gram-negative strain Escherichia coli and to yeast fungus Сandida albicans using agar diffusion method. The medicines of comparison were gentamicin for bacteria and nystatin for yeast fungus Сandida albicans. It has been shown that Polygonum L. essential oils has moderate antibacterial effect to gram-positive microorganisms and weak antifungal activity to Candida albicans yeast fungus. At the second stage of our researches wound healing properties of ointment form of 3% essential oil was researched on the model of flat dermal wounds. To assess the influence of essential oil on healing processes the model of flat dermal wound. The speed of wound healing on rats of different groups was judged based on assessment the area of a wound from time to time. During research of wound healing properties disturbance of integral in neither group: general condition and behavior of animals, food intake, and excretion. Wound healing action of 3% ointment on base of Polygonum L. essential oil and polyethyleneglycol is comparable with the action of reference substances. As more favorable healing dynamics was observed in the experimental group than in control group, the tested ointment can be deemed more promising for further detailed study as wound healing means.

Keywords: antibacterial, antifungal, bioactive substances, essential oils, isolation, Polygonum L.

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Authors: Quang Minh Dinh, Jian Guang Qin, Sabine Dittmann, Dinh Dac Tran

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The age and population structure the dermal gopy Parapocryptes serperaster were studied using length distributions, otolith and von Bertalanffy model in the Mekong Delta over a whole year through monthly sampling. The sex ratio of P. serperaster was near 1:1, and von Bertalanffy growth parameters were L∞= 25.2 cm, K = 0.74 yr-1, and t0 = -0.22 yr-1. Fish size at first entry to fishery was 14.6 cm, and fishing mortality (1.57 yr-1) and natural mortality (1.51 yr-1) accounted for 51% and 49% of the total mortality (3.07 yr-1), respectively. Relative yield-per-recruit and biomass-per-recruit analyses revealed the levels of maximum exploitation yield (Emax = 0.83), maximum economic yield (E0.1 = 0.71) and the yield at 50% reduction of exploitation (E0.5 = 0.37). Otoliths from 164 female and 196 male gobies were readable, and the otolith morphometry data were used for age identification. The mean age estimated by reading otolith annual rings and by analysing length frequency distribution was consistent. This study shows that the otolith morphometry is a reliable method for aging this goby and possibly also applicable for other tropical gobies. The fishery analysis indicates that this goby stock has not been overexploited in the Mekong Delta.

Keywords: Parapcryptes serperaster, otolith, age, pulation structure, Vietnam

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Authors: Ehsan A. Yahia, Ayman E. El-Sharkawey, Magda M. Bayoumi

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Background: Wound dressings perform a crucial role in cutaneous wound management due to their ability to protect wounds and promote dermal and epidermal tissue regeneration. Aim: To evaluate the effectiveness of using hydrogel/nano silver-based dressing vs. traditional dressing on diabetic foot wound healing. Methods: Sixty patients with type-2 diabetes hospitalized for diabetic foot wound treatment were recruited from selected Surgical departments. A prospective randomized control study was carried. Results: The results showed that the percentage of a reduction rate of the ulcer by the third week of the treatment in the hydrogel/nano silver-based dressing group was higher (15.11%) than in the traditional wound dressing group (33.44%). Moreover, the mean ulcer size "sq mm" in the hydrogel/nano silver-based dressing group recognized a faster healing rate (15.11±7.89) and considerably lesser in comparison to the traditional in the third week (21.65±8.4). Conclusion: The hydrogel/nanosilver-based dressing showed better results than traditional dressing in managing diabetic ulcer foot.

Keywords: diabetes, wound care, diabetic foot, wound dressing, hydrogel nanosilver

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Authors: Amir Feily, Hazhir Shahmoradi Akram, Mojtaba Ghaedi, Farshid Javdani, Naser Hatami, Navid Kalani, Mohammad Zarenezhad

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Background: Filler-induced facial skin ischemia is a potential complication of dermal filler injections that can result in tissue damage and necrosis. Nitroglycerin has been suggested as a treatment option due to its vasodilatory effects, but its efficacy in this context is unclear. Methods: A narrative review was conducted to examine the available evidence on the efficacy of nitroglycerin in filler-induced facial skin ischemia. Relevant studies were identified through a search of electronic databases and manual searching of reference lists. Results: The review found limited evidence supporting the efficacy of nitroglycerin in this context. While there were case reports where the combination of nitroglycerin and hyaluronidase was successful in treating filler-induced facial skin ischemia, there was only one case report where nitroglycerin alone was successful. Furthermore, a rat model did not demonstrate any benefits of nitroglycerin and showed harmful results. Conclusion: The evidence regarding the efficacy of nitroglycerin in filler-induced facial skin ischemia is inconclusive and seems to be against its application. Further research is needed to determine the effectiveness of nitroglycerin alone and in combination with other treatments for this condition. Clinicians should consider limited evidence bases when deciding on treatment options for patients with filler-induced facial skin ischemia.

Keywords: nitroglycerin, facial, skin ischemia, fillers, efficacy, narrative review

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Authors: Zayre I. Gonzalez-Acevedo, Marco A. Garcia-Zarate

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Environmental pollution by heavy metals is due to several processes, whether natural as weathering, or anthropogenic, related to human activities. Geysers may content dissolved heavy metals, related with their geothermal origin, and they are widely used by local people and tourists for treatment of dermal diseases and other therapeutic applications. In this study, 20 geysers with temperatures between 32 to 94 °C, located in the vicinity of Queretaro and Guanajuato in Central Mexico, were studied. These geysers were sampled in dry and rainy seasons in order to investigate seasonal changes of trace elements. The samples were analyzed in SWAMP Lab, University of Alberta, Canada for 34 elements. Most of the analyzed trace elements sowed concentrations below guidelines for natural waters. The elements showed seasonal variability with higher concentrations during rainy season. Arsenic varied from 49.29 to 2.16 µg L⁻¹. Arsenic was highly correlated with Fe, Sr, Th and Tl. Barium varied from 93.52 to 1.79 µg L⁻¹. Barium was highly correlated with Co, Cr, Mo, Ni, U, V, and Y. Copper and Zinc were correlated as well. According to the comparison of sites and the correlations between trace elements, their source was identified as natural regional, geothermal or anthropogenic origin. Because of application of geyser's water to balneology and health treatments, and also, because they are located in an urban zone in development, advise on their direct uses, according to their environmental quality is appointed in this research.

Keywords: balneology, direct uses, environmental quality and trace elements

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Authors: Kwanputtha Arunprasert, Chaiyakarn Pornpitchanarong, Praneet Opanasopit, , Prasopchai Patrojanasophon

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Capsaicin, a recently FDA-approved drug for the topical treatment of neuropathic pain, is associated with several side effects like burning sensation and erythema leading to severe skin irritation and poor patient compliance. These unwanted side effects are due to the rapid penetration of capsaicin into the epidermis and low permeation to the dermis layer. The purpose of this study was to develop nanostructured lipid carriers (NLCs) that entrapped capsaicin for reducing dermal irritation. Solid lipid (glyceryl monostearate (GM), cetyl palmitate (CP), cetyl alcohol (COH), stearic acid (SA), and stearyl alcohol (SOH)) and surfactant (Tween®80, Tween®20, and Span®20) were varied to obtained optimal capsaicin-loaded NLCs. The formulation using CP as solid lipid and Tween®80 as a surfactant (F2) demonstrated the smallest size, excellent colloidal stability, and narrow range distribution of the particles as being analyzed using Zetasizer. The obtained capsaicin-loaded NLCs were then characterized by entrapment efficiency (EE) and loading capacity (LC). The release characteristics followed Higuchi kinetics, and the prolonged capsaicin release may result in the reduction in skin irritation. These results could demonstrate the potentials of capsaicinloaded lipid-based nanoparticles for topical drug delivery.

Keywords: capsaicin, lipid-based nanoparticles, nanostructured lipid carriers, topical drug delivery system

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Authors: H. Pegram, R. Stevens, L. De Girolamo

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Aligned electrospun nanofibres act as effective neuronal and glial cell scaffolds that can be layered to contain multiple sheets harboring different cell populations. This allows personalised biofunctional prostheses to be manufactured with both acellular and cellularised layers for the treatment of spinal cord injury. Additionally, the manufacturing route may be configured to produce in-vitro 3D cell based model of spinal cord injury to aid drug development and enhance prosthesis performance. The goal of this investigation was to optimise the multi-layer scaffold design parameters for prosthesis manufacture, to enable the development of multi-layer patient specific implant therapies. The work has also focused on the fabricating aligned nanofibre scaffolds that promote in-vitro neuronal and glial cell population growth, cell-to-cell interaction and long-term survival following trauma to mimic an in-vivo spinal cord lesion. The approach has established reproducible lesions and has identified markers of trauma and regeneration marked by effective neuronal migration across the lesion with glial support. The investigation has advanced the development of an in-vitro model of traumatic spinal cord injury and has identified a route to manufacture prostheses which target the repair spinal cord injury. Evidence collated to investigate the multi-layer concept suggests that physical cues provided by nanofibres provide both a natural extra-cellular matrix (ECM) like environment and controls cell proliferation and migration. Specifically, aligned nanofibre layers act as a guidance system for migrating and elongating neurons. On a larger scale, material type in multi-layer systems also has an influence in inter-layer migration as cell types favour different material types. Results have shown that layering nanofibre membranes create a multi-level scaffold system which can enhance or prohibit cell migration between layers. It is hypothesised that modifying nanofibre layer material permits control over neuronal/glial cell migration. Using this concept, layering of neuronal and glial cells has become possible, in the context of tissue engineering and also modelling in-vitro induced lesions.

Keywords: electrospinning, layering, lesion, modeling, nanofibre

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Authors: Narine Ghazaryan, Joana Catarina Macedo, Sara Vaz, Naira Ayvazyan, Elsa Logarinho

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Macrovipera lebetina obtusa (MLO) is a poisonous snake in Armenia. Obtustatin represents the shortest known monomeric disintegrin, isolated from the snake venom of MLO, and is known to specifically inhibit α1β1 integrin. Its oncostatic effect is due to the inhibition of angiogenesis, which likely arises from α1β1 integrin inhibition in the endothelial cells. To explore the therapeutic potential of the MLO snake venom and obtustatin, we studied activity of obtustatin and MLO venom in vitro, by testing their efficacy in human dermal microvascular endothelial cells (HMVEC-D) and in vivo, using chick embryo chorioallantoic membrane assay (CAM assay). Our in vitro results showed that obtustatin in comparison with MLO venom did not exhibit cytotoxic activity in HMVEC-D cells in comparison to MLO venom. But in vivo results have shown that 4µg /embryo (90 µM) of obtustatin inhibited angiogenesis induced by FGF2 by 17% while MLO snake venom induced 22% reduction of the angiogenic index. The concentration of obtustatin in the crude MLO venom was 0.3 nM, which is 300.000 times less than the concentration of the obtustatin itself. Given this enormous difference in concentration, it is likely that some components of the crude venom contribute to the observed anti-angiogenic effect. Hypotheses will be ascertained to justify this action: components in the MLO venom may increase obtustatin efficacy or have independent but synergic anti-angiogenic activities.

Keywords: angiogenesis, alpa1 beta 1 integrin, Macrovipera lebetina obtusa, obtustatin

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Authors: Hadi Rad

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Unrestricted somatic stem cells (USSCs) loaded in nanofibrous PHBV scaffold can be used for skin regeneration when grafted into full-thickness skin defects of rats. Nanofibrous PHBV scaffolds were designed using electrospinning method and then, modified with the immobilized collagen via the plasma method. Afterward, the scaffolds were evaluated using scanning electron microscopy, physical and mechanical assays. In this study; nanofibrous PHBV scaffolds loaded with and without USSCs were grafted into the skin defects. The wounds were subsequently investigated at 21 days after grafting. Results of mechanical and physical analyses showed good resilience and compliance to movement as a skin graft. In animal models; all study groups excluding the control group exhibited the most pronounced effect on wound closure, with the statistically significant improvement in wound healing being seen on post-operative Day 21. Histological and immunostaining examinations of healed wounds from all groups, especially the groups treated with stem cells, showed a thin epidermis plus recovered skin appendages in the dermal layer. Thus, the graft of collagen-coated nanofibrous PHBV scaffold loaded with USSC showed better results during the healing process of skin defects in rat model.

Keywords: collagen, nanofibrous PHBV scaffold, unrestricted somatic stem cells, wound healing.

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Authors: Sheimah El Bejjaji, Lara Gorsek, Chandler Quilchez, Joaquim Suñer, Mireia Mallandrich

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Flurbiprofen is an anti-inflammatory drug used in several treatments. The purpose of this study was to compare the permeation of two different formulations of flurbiprofen through the human skin. The first formulation was a solution of flurbiprofen dissolved with polyethylene glycol 3350 (PEG 3350). The second formulation was flurbiprofen encapsulated in poly-ɛ-caprolactone (PɛCL) nanoparticles (NPs), stabilized with poloxamer 188, submitted individually for freeze-drying with PEG 3350 as a cryoprotectant and sterilized by gamma-irradiation. Human skin was obtained from the abdominal region of a healthy patient. The experimental protocol was approved by the Bioethics Committee of Barcelona SCIAS Hospital (Spain), and they obtained the written informed consent forms. After being frozen to -20ºC, the skin samples were cut with a dermatome at 400 µm. The ex vivo permeation study was performed in Franz diffusion cells with a diffusion area of 2.54 cm². Skin samples were placed between two compartment sites, the dermal side in contact with the receptor medium and the epidermis side in contact with the donor chamber to which the formulation was applied. The permeation study was conducted for 24 hours at 32 ± 0.5 °C in accordance with sink conditions. The results were analyzed with an unpaired t-test, and the p-values indicate the formulation with nanoparticles had a higher permeability coefficient, flux, partition parameter, diffusion parameter, and lag time. The applicability of this formulation topically can benefit articulations and ligament inflammation as an alternative to oral drugs.

Keywords: anti-inflammatory drug, flurbiprofen, human skin, nanoparticles, skin permeation

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Authors: Luay Alsalmi

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Background: Facial filler is the most common type of cosmetic surgery next to botox. Permanent filler is preferred nowadays due to the low cost brought about by non-recurring injection appointments. However, such fillers pose a higher risk for complications, with even greater adverse effects when the procedure is done using unknown dermal filler injections. AIM: This study aimed to establish an algorithm to categorize and manage patients that receive permanent fillers. Materials and Methods: Twelve participants were presented to the service through emergency or as outpatient from November 2015 to May 2021. Demographics such as age, sex, date of injection, time of onset, and types of complications were collected. After examination, all cases were managed based on an algorithm established. FACE-Q was used to measure overall satisfaction and psychological well-being. Results: The algorithm to diagnose and manage these patients effectively with a high satisfaction rate was established in this study. All participants were non-smoker females with no known medical comorbidities. The algorithm presented determined the treatment plan when faced with complications. Results revealed high appearance-related psychosocial distress was observed prior to surgery, while it significantly dropped after surgery. FACE-Q was able to establish evidence of satisfactory ratings among patients prior to and after surgery. Conclusion: This treatment algorithm can guide the surgeon in formulating a suitable plan with fewer complications and a high satisfaction rate.

Keywords: facial filler, FACE-Q, psycho-social stress, botox, treatment algorithm

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Authors: Ben Abderrahmane Ayoub El Fateh, Bnina Rachid

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A comprehensive comparison of antioxidant defenses in the dermis and epidermis and their response to exposure to ultraviolet (UV) irradiation has not previously been attempted. In this study, enzymic and non-enzymic antioxidants in epidermis and dermis of hairless mice were compared. Enzyme activities are presented both as units/gram of skin and units/milligram of protein; arguments are presented for the superiority of skin wet weight as a reference base. Catalase, glutathione peroxidase, and glutathione reductase (units/gram of skin) were higher in the epidermis than dermis by 49%, 86%, and 74%, respectively. Superoxide dismutase did not follow this pattern. Lipophilic antioxidants ( -tocopherol, ubiquinol 9, and ubiquinone 9) and hydrophilic antioxidants (ascorbic acid, dehydroascorbic acid, and glutathione) were 24–95% higher in the epidermis than in dermis. In contrast, oxidized glutathione was 60% lower in the epidermis than in dermis. Mice were irradiated with solar light to examine the response of these cutaneous layers to UV irradiation. After irradiation with 25 J/cm2 (UVA + UVB, from a solar simulator), 10 times the minimum erythemal dose, epidermal and dermal catalase and superoxide dismutase activities were greatly decreased. Tocopherol, ubiquinol 9, ubiquinone 9, ascorbic acid, dehydroascorbic acid, and reduced glutathione decreased in both epidermis and dermis by 26-93%. Oxidizedgiutathione showed a slight, non-significant increase. Because the reduction in total ascorbate and catalase was much more severe in the epidermis than dermis, it can be concluded that UV light is more damaging to the antioxidant defenses in the epidermis than in the dermis.

Keywords: antioxidant defenses, enzymic, epidermis, oxidizedgiutathione

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Authors: Thodoris Tsagkaris, Marios Stavropoulos, Panagiotis Theodosis-Nobelos, Charalampos Triantis

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Isotretinoin is a widely used therapeutic for the treatment of acne vulgaris and various other skin disorders. However, since its approval, many side effects and contraindications have been described, particularly important, such as teratogenicity as well as liver disease and dermal deterioration. In a very important allegation, isotretinoin has been linked with psychiatric symptoms like depression, suicidal ideation, schizophrenia, and hypervitaminosis A syndrome characteristics. These adverse effects have raised significant concerns regarding the safety of isotretinoin. Numerous studies and research have associated isotretinoin with side effects on the mental health of patients and have proposed plausible mechanisms regarding this suspected causative relationship. However, the evidence is still contradicting, and the data disperse, making their validity less valuable. Thus, in the present study, we aim to analyze further the available literature and present a complete analysis of the side effects of isotretinoin, with particular emphasis on the effects it may have on the mental health of patients. The review is based on international articles from broad scientific electronic databases like PubMed and Scopus. This review concludes that although many studies have associated isotretinoin with mental effects like depression, bipolar disorder, schizophrenia, and suicidal ideation, the data are still insufficient and often contradictory. In fact, additional studies with accurate data and larger double-blinded samples, and more analytic systematic reviews are required. It is especially important to monitor the dose and the intervals that isotretinoin has to be administered in order to potentially cause mental health problems, as well as the duration of treatment and the role that the patient's medical and pharmaceutical history may play.

Keywords: acne, depression, isotretinoin, mental health

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Authors: Anissa Mary Thomas Thattil, Nancy Angeline Gnanaselvam, B. Ramakrishna Goud

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Faecal sludge management (FSM) is an unorganized sector, and in India, there is an absence of regulations regarding the collection, transport, treatment, and disposal of faecal sludge. FSM has a high degree of occupational hazards that need to be thoroughly understood in order to shape effective solutions. On-field observations of five FSM operations were conducted in Anekal Taluk of southern Karnataka. All five of the FSM operations were privately owned and snowball method of sampling was employed. Two types of FS operations observed were: mechanical emptying involving direct human contact with faecal sludge and mechanical emptying without direct human contact with faecal sludge. Each operation was manned by 3-4 faecal sludge operators (FSOs). None of the observed FSOs used personal protective equipment. According to the WHO semi-quantitative risk assessment, the very high risk occupational hazards identified were dermal contact with faecal sludge, inhalation of toxic gases, and social stigma. The high risk hazards identified were trips and falls, injuries, ergonomic hazards, substance abuse, and mental health problems. In all five FSM operations, the collected faecal sludge was discharged untreated onto abandoned land. FSM in India is fraught with occupational and environmental hazards which need to be urgently addressed. This includes formalizing the institution of FSM, contextualized behaviour change communication, capacity building of local bodies, awareness programmes among agriculturists and FSOs, and designation of sites for the safe harnessing of faecal sludge as soil nutrient.

Keywords: faecal sludge, faecal sludge management, FSM, occupational hazards, sanitation

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Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Dobdin, Anatoly Skripal, Andrey Usanov, Dmitry Usanov

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One of the most important unsolved problems in modern medicine is the increase of chronic diseases that lead to organ dysfunction or even complete loss of function. Current methods of treatment do not result in decreased mortality and disability statistics. Currently, the best treatment for many patients is still transplantation of organs and/or tissues. Therefore, finding a way of correct artificial matrix biofabrication in case of limited number of natural organs for transplantation is a critical task. One important problem that needs to be solved is development of a nondestructive and noninvasive method to analyze dynamic changes of mechanical characteristics of a matrix with minimal side effects on the growing cells. This research was focused on investigating the properties of matrix as a marker of graft condition. In this study, the collagen gel with human primary dermal fibroblasts in suspension (60, 120, 240*103 cells/mL) and collagen gel with cell spheroids were used as model objects. The stiffness and elasticity characteristics were evaluated by a semiconductor laser autodyne. The time and cell concentration dependency of the stiffness and elasticity were investigated. It was shown that these properties changed in a non-linear manner with respect to cell concentration. The maximum matrix stiffness was observed in the collagen gel with the cell concentration of 120*103 cells/mL. This study proved the opportunity to use the mechanical properties of matrix as a marker of graft condition, which can be measured by noninvasive semiconductor laser autodyne technique.

Keywords: graft, matrix, noninvasive method, regenerative medicine, semiconductor laser autodyne

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Authors: Samir El-Gendy, Doaa Zaghloul

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The current study dealt with the gross and microscopic anatomy of the integument of male ostrich in addition to the histological features of different areas of skin by light and SEM. The ostrich skin is characterized by prominent feather follicles and bristles. The number of feather follicles was determined per cm2 in different regions. The integument of ostrich had many modifications which appeared as callosities and scales, nail and toe pads. They were sternal, pubic and Achilles tendon callosities. The vacuolated epidermal cells were seen mainly in the skin of legs and to a lesser extent in the skin of back and Achilles areas. Higher lipogenic potential was expressed by epidermis from glabrous areas of ostrich skin. The dermal papillae were found in the skin of feathered area of neck and back and this was not a common finding in bird's skin which may give resistance against shearing forces in these regions of ostrich skin. The thickness of the keratin layer of ostrich varied, being thick and characteristically loose in the skin at legs, very thin and wavy at neck, while at Achilles skin area, scale and toe pad were thick and more compact, with the thickest very dense and wavy keratin layer at the nail. The dermis consisted of superficial layer of dense irregular connective tissue characterized by presence of many vacuoles of different sizes just under the basal lamina of the epithelium of epidermis and deep layer of dense regular connective tissue. This result suggested presence of fat droplets in this layer which may be to overcome the lack of good barrier of cutaneous water loss in epidermis.

Keywords: ostrich, light microscopy, scanning electron microscopy, integument, skin modifications

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Authors: Alyaa Abdlwahab

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Cutaneous leishmaniasis represents a serious health problem in Syria; this problem has become noticeably aggravated after the civil war in the country. Leishmania tropica parasite is the main cause of cutaneous leishmaniasis in Syria. In order to control the disease, we need an effective vaccine against leishmania parasite. DNA vaccination remains one of the favorable approaches that have been used to face cutaneous leishmaniasis. Ribosomal protein S4 is responsible for important roles in Leishmania parasite life. DNA vaccine based on S4 gene has been used against infections by many species of Leishmania parasite but leishmania tropica parasite, so this gene represents a good candidate for DNA vaccine construction. After proving the existence of ribosomal protein S4 gene in a Syrian strain of Leishmania tropica (LCED Syrian 01), sequencing it and cloning it into pCI plasmid, BALB/C mice were inoculated with pCI-S4 DNA vaccine. The immune response was determined by monitoring the lesion progression in inoculated BALB/C mice for six weeks after challenging mice with Leishmania tropica (LCED Syrian 01) parasites. IL-12, IFN-γ, and IL-4 were quantified in draining lymph nodes (DLNa) of the immunized BALB/C mice by using the RT-qPCR technique. The parasite burden was calculated in the final week for the footpad lesion and the DLNs of the mice. This study proved the existence and the expression of the ribosomal protein S4 gene in Leishmania tropica (LCED Syrian 01) promastigotes. The sequence of ribosomal protein cDNA S4 gene was determined and published in Genbank; the gene size was 822 bp. Expression was also demonstrated at the level of cDNA. Also, this study revealed that pCI-S4 DNA vaccine induces TH1\TH2 response in immunized mice; this response prevents partially developing a dermal lesion of Leishmania.

Keywords: ribosomal protein S4, DNA vaccine, Leishmania tropica, BALB\c

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Authors: Abhay Asthana, Shally Toshkhani, Gyati Shilakari

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The present research was aimed to develop a biodegradable dermal patch formulation for wound healing in a novel, sustained and systematic manner. The goal is to reduce the frequency of dressings with improved drug delivery and thereby enhance therapeutic performance. In present study optimized formulation was designed using component polymers and excipients (e.g. Hydroxypropyl methyl cellulose, Ethylcellulose, and Gelatin) to impart significant folding endurance, elasticity and strength. Gelatin was used to get a mixture using ethylene glycol. Chitosan dissolved in suitable medium was mixed with stirring to gelatin mixture. With continued stirring to the mixture Curcumin was added in optimized ratio to get homogeneous dispersion. Polymers were dispersed with stirring in final formulation. The mixture was sonicated casted to get the film form. All steps were carried out under under strict aseptic conditions. The final formulation was a thin uniformly smooth textured film with dark brown-yellow color. The film was found to have folding endurance was around 20 to 21 times without a crack in an optimized formulation at RT (23C). The drug content was in range 96 to 102% and it passed the content uniform test. The final moisture content of the optimized formulation film was NMT 9.0%. The films passed stability study conducted at refrigerated conditions (4±0.2C) and at room temperature (23 ± 2C) for 30 days. Further, the drug content and texture remained undisturbed with stability study conducted at RT 23±2C for 45 and 90 days. Percentage cumulative drug release was found to be 80% in 12 h and matched the biodegradation rate as drug release with correlation factor R2 > 0.9. The film based formulation developed shows promising results in terms of stability and release profiles.

Keywords: biodegradable, patch, bioactive, polymer

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Authors: Huixin Wei, Shibin Wang, Linan Li, Lei Zhou, Xinhao Tu

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Facial skin is a biomedical material with complex mechanical properties of anisotropy, viscoelasticity, and hyperelasticity. The mechanical properties of facial skin are crucial for a number of applications including facial plastic surgery, animation, dermatology, cosmetic industry, and impact biomechanics. Skin is a complex multi-layered material which can be broadly divided into three main layers, the epidermis, the dermis, and the hypodermis. Collagen fibers account for 75% of the dry weight of dermal tissue, and it is these fibers which are responsible for the mechanical properties of skin. Many research on the anisotropic mechanical properties are mainly concentrated on in vitro, but there is a great difference between in vivo and in vitro for mechanical properties of the skin. In this study, we presented a method to measure the mechanical properties of facial skin in vivo. Digital image correlation (DIC) and indentation tests were used to obtain the experiment data, including the deformation of facial surface and indentation force-displacement curve. Then, the experiment was simulated using a finite element (FE) model. Application of Computed Tomography (CT) and reconstruction techniques obtained the real tissue geometry. A three-dimensional FE model of facial skin, including a bi-layer system, was obtained. As the epidermis is relatively thin, the epidermis and dermis were regarded as one layer and below it was hypodermis in this study. The upper layer was modeled as a Gasser-Ogden-Holzapfel (GOH) model to describe hyperelastic and anisotropic behaviors of the dermis. The under layer was modeled as a linear elastic model. In conclusion, the material properties of two-layer were determined by minimizing the error between the FE data and experimental data.

Keywords: facial skin, indentation test, finite element, digital image correlation, computed tomography

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Authors: Piyachat Chuysinuan, Nitirat Chimnoi, Arthit Makarasen, Nanthawan Reuk-Ngam, Pitt Supaphol, Supanna Techasakul

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In this work, biomaterial wound dressings was developed based on gelatin containing herbal substances (essential oil), a substance from the plant Eupatorium adenophorum Spreng (Crofton weed) that used as traditional wound healers. Gelatin hydrogel was prepared from a 10 wt-% gelatin solution. The oil in water (o/w) emulsion Eupatorium adenophorum of essential oil were prepared and used Pluronic F68 as a surfactant. The 10, 20, and 30 % v/v emulsion were mixed with gelatin solution and cast into film. These hydrogels were tested for their gel fraction, swelling and weight loss behavior. With an increase in the emulsion concentration the emulsion-loaded in hydrogels, the gel fraction were decreased due to the crosslink density, while the swelling and weight loss behavior were increased with an increasing in the emulsion content. The potential to use the emulsion-containing gelatin hydrogels as wound dressing was assessed on investigation the release characteristics of the as-loaded hydrogels. The E. adenophorum essential oil was first identified the chemical composition by using GC-MS analysis. The principal components of the oil were p-cymene (16.23%), bornyl acetate (11.84%), and amorpha-4, 7(11)-diene (10.51%). The hydrogel wound dressing containing essential oil was then characterized for their antibacterial activity against Gram-positive and Gram-negative in order to elucidate their potential for use as antibacterial wound dressings by using agar disk diffusion methods. The result showed that E. adenophorum essential oil and the emulsion-loaded gelatin hydrogel inhibited the growth of the test pathogens, Staphylococcus aureus and Staphylococcus epidermidis and increased with increasing the initial amount of essential oil in the hydrogels which confirmed their application as antibacterial wound dressings. Furthermore, the potential use of these wound dressings was further assessed in terms of the indirect cytotoxicity, in vitro attachment and proliferation of dermal human fibroblasts cultured in the hydrogel wound dressings.

Keywords: hydrogel, antibacterial wound dressing, Eupatorium adenophorum essential oil, gelatin

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Authors: Genesis Julyus T. Agcaoili, Esperanza C. Cabrera

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Thirty (30) isolates of lactic acid bacteria (LAB) from traditionally-prepared fermented products specifically fermented soy-bean paste, fermented mustard and fermented rice-fish mixture were studied for their in vitro antimicrobial and cytotoxic activities. Seventeen (17) isolates were identified as Lactobacillus plantarum, while 13 isolates were identified as Enterococcus spp using 16s rDNA sequences. Disc diffusion method was used to determine the antibacterial activity of LAB against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), while the modified agar overlay method was used to determine the antifungal activity of LAB isolates on the yeast Candida albicans, and the dermatophytes Microsporum gypseum, Trichophyton rubrum and Epidermophyton floccosum. The filter-sterilized LAB supernatants were evaluated for their cytotoxicity to mammalian colon cancer cell lines (HT-29 and HCT116) and normal human dermal fibrolasts (HDFn) using resazurin assay (PrestoBlueTM). Colchicine was the positive control. No antimicrobial activity was observed against the bacterial test organisms and the yeast Candida albicans. On the other hand, all of the tested LAB strains were fungicidal for all the test dermatophytes. Cytotoxicity index profiles of the supernatants of the 15 randomly picked LABs and negative control (brain heart infussion broth) suggest nontoxicity to the cells when compared to colchicine, whereas all LAB supernatants were found to be cytotoxic to HT-29 and HCT116 colon cancer cell lines. Results provide strong support for the role of the lactic acid bacteria studied in antimicrobial treatment and anticancer therapy.

Keywords: antimicrobial, fermented products, fungicidal activity, lactic acid bacteria, probiotics

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Authors: Ginger Egberts, Marianne Schaaphok, Fred Vermolen, Paul van Zuijlen

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A few years ago, a promising morphoelastic model was developed for the simulation of contraction formation after burn injuries. Contraction can lead to a serious reduction in physical mobility, like a reduction in the range-of-motion of joints. If this is the case in a healing burn wound, then this is referred to as a contracture that needs medical intervention. The morphoelastic model consists of a set of partial differential equations describing both a chemical part and a mechanical part in dermal wound healing. These equations are solved with the numerical finite element method (FEM). In this method, many calculations are required on each of the chosen elements. In general, the more elements, the more accurate the solution. However, the number of elements increases rapidly if simulations are performed in 2D and 3D. In that case, it not only takes longer before a prediction is available, the computation also becomes more expensive. It is therefore important to investigate alternative possibilities to generate the same results, based on the input parameters only. In this study, a surrogate neural network has been designed to mimic the results of the one-dimensional morphoelastic model. The neural network generates predictions quickly, is easy to implement, and there is freedom in the choice of input and output. Because a neural network requires extensive training and a data set, it is ideal that the one-dimensional FEM code generates output quickly. These feed-forward-type neural network results are very promising. Not only can the network give faster predictions, but it also has a performance of over 99%. It reports on the relative surface area of the wound/scar, the total strain energy density, and the evolutions of the densities of the chemicals and mechanics. It is, therefore, interesting to investigate the applicability of a neural network for the two- and three-dimensional morphoelastic model for contraction after burn injuries.

Keywords: biomechanics, burns, feasibility, feed-forward NN, morphoelasticity, neural network, relative surface area wound

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Authors: Hardisiswo Soedjana, Inne Caroline

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Deep dermal or full thickness burns are inevitably lead to post-burn contractures. These contractures remain to be one of the most concerning late complications of burn injuries. Surgical management includes releasing the contracture followed by resurfacing the defect accompanied by post-operative rehabilitation. Optimal treatment of post-burn contractures depends on the characteristics of the contractures. This study is aimed to describe clinical characteristics, problems, and management of post-burn contractures in adults and children. A retrospective analysis was conducted from medical records of patients suffered from contractures after burn injuries admitted to Hasan Sadikin general hospital between January 2016 and January 2018. A total of 50 patients with post burn contractures were included in the study. There were 17 adults and 33 children. Most patients were male, whose age range within 15-59 years old and 5-9 years old. Educational background was mostly senior high school among adults, while there was only one third of children who have entered school. Etiology of burns was predominantly flame in adults (82.3%); whereas flame and scald were the leading cause of burn injury in children (11%). Based on anatomical regions, hands were the most common affected both in adults (35.2%) and children (48.5%). Contractures were identified in 6-12 months since the initial burns. Most post-burn hand contractures were resurfaced with full-thickness skin graft (FTSG) both in adults and children. There were 11 patients who presented with recurrent contracture after previous history of contracture release. Post-operative rehabilitation was conducted for all patients; however, it is important to highlight that it is still challenging to control splinting and exercise when patients are discharged and especially the compliance in children. In order to improve quality of life in patients with history of deep burn injuries, prevention of contractures should begin right after acute care has been established. Education for the importance of splinting and exercise should be administered as comprehensible as possible for adult patients and parents of pediatric patients.

Keywords: burn, contracture, education, exercise, splinting

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Authors: Snezana Rajkovic, Miroslava Markovic, Ljubinko Rakonjac, Aleksandar Lucic, Radoslav Rajkovic

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Woolly apple aphid, Eriosoma lanigerum (Hausmann) is a widely distributed pest of apple trees, especially where its parasites have been killed by insecticides. It can also be found on pear, hawthorn, mountain ash, and elm trees. Relatively small to medium-sized aphids, characterized by a reddish-brown body, a blood-red stain when crushed and a fluffy, flocculent wax covering. Specialized dermal glands produce the characteristic fluffy or powdery wax, which gives E. lanigerum its characteristic 'woolly' appearance. Also, woolly apple aphid is a problemm in nursery production of spure.The experiments were carried out in the nursery “Nevade” in Gornji Milanovac, "Srbijasume" on the spruce seedlings, aged 2 years. In this study, organic insecticide King Bo, aqueous solution (a. i. oxymatrine 0.2% + psoralen 0.4%), manufacturer Beijing Kingbo Biotech Co. Ltd., Beijing, China. extracted from plants and used as pesticides in nursery production were investigated. King Bo, bioinsecticide is manufactured from refined natural herbal extract several wild medicinal plants, such as Sophora flavescens Ait, Veratrum nigrum L, A. Carmichael, etc. Oxymatrine 2.4 SL is a stomach poison that has antifeeding and repellent action. This substance stimulates development and growth in a host plant and also controls the appearance of downy mildew.The trials were set according to instructions of methods-monitoring of changes in the number of larvae and adults compared to before treatment. The treatment plan was made according to fully randomized block design. The experiment was conducted in four repetitions. The basic plot had the area of 25 m2. Phytotoxicity was estimated by PP methods 1/135 (2), the intensity of infection according to Towsend-Heuberger, the efficiency by Abbott, the analysis of variance with Ducan test and PP/181 (2).

Keywords: bioinsecticide, efficacy, nurssery production, woolly apple aphid

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Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

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Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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Authors: Yemima Dani Riani, Anggraini Barlian

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Green turtle (Chelonia mydas) is one of well known long-lived turtle. Its age can reach 100 years old. Senescence in green turtle is an interesting process to study because until now no clear explanation has been established about senescence at cellular or molecular level in this species. Since 1999, green turtle announced as an endangered species. Hence, establishment of fibroblast skin cell culture of green turtle may be material for future study of senescence. One common marker used for detecting senescence is telomere shortening. Reduced telomerase activity, the reverse transcriptase enzyme which adds TTAGGG DNA sequence to telomere end, may also cause senescence. The purpose of this research are establish and identify green turtle fibroblast skin cell culture and also compare telomere length and telomerase activity from passage 5 and 14. Primary cell culture made with primary explant method then cultured in Leibovitz-15 (Sigma) supplemented by 10% Fetal Bovine Serum (Sigma) and 100 U/mL Penicillin/Streptomycin (Sigma) at 30 ± 1oC. Cells identified with Rabbit Anti-Vimentin Polyclonal Antibody (Abcam) and Goat Polyclonal Antibody (Abcam) using confocal microscope (Zeiss LSM 170). Telomere length obtained using TeloTAGGG Telomere Length Assay (Roche) while telomerase activity obtained using TeloTAGGG Telomerase PCR ElisaPlus (Roche). Primary cell culture from green turtle skin had fibroblastic morphology and immunocytochemistry test with vimentin antibody proved the culture was fibroblast cell. Measurement of telomere length and telomerase activity showed that telomere length and telomerase activity of passage 14 was greater than passage 5. However, based on morphology, green turtle fibroblast skin cell culture showed senescent morphology. Based on the analysis of telomere length and telomerase activity, suspected fibroblast skin cell culture of green turtles is not undergo aging through telomere shortening.

Keywords: cell culture, chelonia mydas, telomerase, telomere, senescence

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Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

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The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

Procedia PDF Downloads 79