In-vitro & in-vivo study of primary epidermal auto, allo and xenografts at different cell passages transplanted on burn wound in oryctolagus cuniculus

The goal of this study was to investigate the wound healing potential using primary epidermal auto-, allo-, and xenografts in rabbit ( Oryctolagus cuniculus). In this study, rabbit and rat skins were harvested and cultured in vitro. Cultured epithelial autograft, allograft and xenograft cells were sprayed onto the three freshly created wounds with one wound acting as a control. The wounds were monitored every two days for four weeks. After four weeks, the rabbits were euthanized; skin biopsies were taken from each healed wound and subjected to hematoxylin and eosin staining followed by immunohistochemical staining using Pancytokeratin, PCK-26 (Abeam, Germany) and Cytokeratin A1/A3 antibodies. All the examined grafts showed favorable healing outcomes because the wounds appeared similar to normal skin upon healing. The only observed difference was related to the thickness of the epidermis layer in the case of the xenograft transplant, which was thinner compared with the autograft and allograft. It was also noted that the healing rates of the wounds treated with the xenograft was similar to wounds treated with autograft and allografts. The level of antigenicity of the 3 grafts appeared to be the same, which is displayed by the similar healing rate of the grafts

Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)

Secretory factors of adipose-derived stem cells have paracrine effects that may have potential in accelerating proliferation of keratinocytes by coculturing both of the cells. This study was conducted to evaluate the indirect effects of ASCs-PCS on the growth of keratinocytes by paracrine activities. Human adipose-derived stem cells and human epidermal keratinocytes were isolated and verified with their specific markers. Distribution and proliferation of ASCs on porous chitosan scaffold were assessed by scanning electron microscope, live/dead assay and Alamar blue assay. Coculture groups were divided into two groups; culture ASCs and culture of ASCs-PCS. Proliferation of keratinocytes in the coculture wells were analysed by Alamar blue assay at 24 hours and 72 hours of coculture while the concentration human epidermal growth factor in culture supernatant were quantified at 72 hours of coculture. Adipose-derived stem cells culture have been established and displayed high expression of CD29, CD73, CD90, CD105 and low expression of CD34. Keratinocytes culture expressed involucrin and cytokeratin 6 as demonstrated by immunocytochemistry. Percentage of proliferation of ASCs within PCS was more than 80% accompanied with spindle-shaped features and good viability. After 72 hours of coculture, the percentage of nHEK in the coculture group was increased due to growth medium. ELISA assay revealed no secretion of EGF from ASCs. This study demonstrated via dual -chamber coculture model, human -derived ASCs and PCS showed no growth stimulatory effect towards the growth of keratinocytes via the tested growth factor, EGF. The proliferation of nHEK in the coculture wells possibly due to other growth factor or other mechanism

Treatment modalities for hyperpigmented skin lesions: A brief overview

Skin hyperpigmentation involves a broad range of skin conditions, including epidermal pigmented lesions, dermal pigmented lesions, and mixed pigmented lesions. Treatment includes various modalities such as brightening cream, chemical peeling, and laser therapy. Responses to various treatment modalities can be quite varied depending on the type of treatment and the degree of pigmentation. Sometimes a lesion can lighten or even partially disappear, while other lesions may recur. This paper provides a brief overview of treatment modalities available for hyperpigmented skin lesions including the importance of photoprotection, various types of brightening creams, suitable types of chemical peels, specific laser therapies targeted for skin hyperpigmentation, and surgery

Effects of Ischemic Preconditioning of Different Intraoperative Ischemic Times of Vascularized Bone Graft Rabbit Models

Background Ischemic preconditioning has been shown to improve the outcomes of hypoxic tolerance of the heart, brain, lung, liver, jejunum, skin, and muscle tissues. However, to date, no report of ischemic preconditioning on vascularized bone grafts has been published. Methods Sixteen rabbits were divided into four groups with ischemic times of 2, 6, 14, and 18 hours. Half of the rabbits in each group underwent ischemic preconditioning. The osteomyocutaneous flaps consisted of the tibia bone, from which the overlying muscle and skin were raised. The technique of ischemic preconditioning involved applying a vascular clamp to the pedicle for 3 cycles of 10 minutes each. The rabbits then underwent serial plain radiography and computed tomography imaging on the first, second, fourth, and sixth postoperative weeks. Following this, all of the rabbits were sacrificed and histological examinations were performed. Results The results showed that for clinical analysis of the skin flaps and bone grafts, the preconditioned groups showed better survivability. In the plain radiographs, except for two non-preconditioned rabbits with intraoperative ischemic times of 6 hours, all began to show early callus formation at the fourth week. The computed tomography findings showed more callus formation in the preconditioned groups for all of the ischemic times except for the 18-hour group. The histological findings correlated with the radiological findings. There was no statistical significance in the difference between the two groups. Conclusions In conclusion, ischemic preconditioning improved the survivability of skin flaps and increased callus formation during the healing process of vascularized bone grafts

Antiproliferative effect of methanolic extraction of tualang honey on human keloid fibroblasts

<p>Abstract</p> <p>Background</p> <p>Keloid is a type of scar which extends beyond the boundaries of the original wound. It can spread to the surrounding skin by invasion. The use of Tualang honey is a possible approach for keloid treatment. The objective of this study was to determine the antiproliferative effect of methanolic extraction of Tualang honey to primary human keloid fibroblasts and to identify the volatile compounds in methanol extraction of Tualang honey.</p> <p>Methods</p> <p>Crude Tualang honey was extracted with methanol and then dried using rota vapor to remove remaining methanol from honey. Normal and keloid fibroblasts were verified and treated with the extracted honey. Cell proliferation was tested with [3-(4,5-dimethylthiazol-2-yi)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] (MTS) assay. Extraction of Tualang honey using methanol was carried out and the extracted samples were analysed using gas chromatography-mass spectrometry (GC-MS). The result was analysed using SPSS and tested with Kruskal-Wallis and Mann-Whitney tests.</p> <p>Results</p> <p>Methanolic extraction of honey has positive anti proliferative effect on keloid fibroblasts in a dose-dependent manner. The presence of fatty acids such as palmitic acid, stearic acid, oleic acid, linoleic acid and octadecanoic acid may contribute to the anti-proliferative effect in keloid fibroblasts.</p> <p>Conclusions</p> <p>The methanolic honey extraction has an antiproliferative effect on keloid fibroblasts and a range of volatile compounds has been identified from Tualang honey. The antiproliferative effect of keloid fibroblasts towards Tualang honey may involve cell signaling pathway. Identifying other volatile compounds from different organic solvents should be carried out in future.</p

Compatibility of Porous Chitosan Scaffold with the Attachment and Proliferation of human Adipose-Derived Stem Cells In Vitro

Adipose-derived stem cells (ASCs) have potential applications in the repair and regeneration of various tissues and organs. The use of various scaffold materials as an excellent template for mimicking the extracellular matrix to induce the attachment and proliferation of different cell types has always been of interest in the field of tissue engineering because ideal biomaterials are in great demand. Chitosan, a marine polysaccharide, have wide clinical applications and it acts as a promising scaffold for cell migration and proliferation. ASCs, with their multi-differentiation potential, and chitosan, with its great biocompatibility with ASCs, were investigated in the present study. ASCs were isolated and were characterized by two different methods: immunocytochemistry and flow cytometry, using the mesenchymal stem cell markers CD90, CD105, CD73 and CD29. The ASCs were then induced to differentiate into adipogenic, osteogenic and chondrogenic lineages. These ASCs were incorporated into a porous chitosan scaffold (PCS), and their structural morphology was studied using a scanning electron microscope and hematoxylin and eosin staining. The proliferation rate of the ASCs on the PCS was assessed using a PrestoBlue viability assay. The results indicated that the PCS provides an excellent template for the adhesion and proliferation of ASCs. Thus, this study revealed that PCS is a promising biomaterial for inducing the proliferation of ASCs, which could lead to successful tissue reconstruction in the field of tissue engineering

Keloid Scarring: Understanding the Genetic Basis, Advances, and Prospects

Keloid disease is a fibroproliferative dermal tumor with an unknown etiology that occurs after a skin injury in genetically susceptible individuals. Increased familial aggregation, a higher prevalence in certain races, parallelism in identical twins, and alteration in gene expression all favor a remarkable genetic contribution to keloid pathology. It seems that the environment triggers the disease in genetically susceptible individuals. Several genes have been implicated in the etiology of keloid disease, but no single gene mutation has thus far been found to be responsible. Therefore, a combination of methods such as association, gene-gene interaction, epigenetics, linkage, gene expression, and protein analysis should be applied to determine keloid etiology

A study on the microbiological and angiogenic properties of burn injuries treated with tualang honey versus silver based dressing

Tualang honey was demonstrated to pose substantial bactericidal as well as bacteriostatic effect. The Aquacel dressing soaked with pure Tualang honey without any dilution was less sticky than manuka and was easier to apply as a dressing