Exogenous peripheral blood mononuclear cells affect the healing process of deep-degree burns
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The regenerative repair of deep‑degree (second degree) burned skin remains a notable challenge in the treatment of burn injury, despite improvements being made with regards to treatment modality and the emergence of novel therapies. Fetal skin constitutes an attractive target for investigating scarless healing of burned skin. To investigate the inflammatory response during scarless healing of burned fetal skin, the present study developed a nude mouse model, which was implanted with normal human fetal skin and burned fetal skin. Subsequently, human peripheral blood mononuclear cells (PBMCs) were used to treat the nude mouse model carrying the burned fetal skin. The expression levels of matrix metalloproteinase (MMP)‑9 and tissue inhibitor of metalloproteinases (TIMP)‑1 were investigated during this process. In the present study, fetal skin was subcutaneously implanted into the nude mice to establish the murine model. Hematoxylin and eosin staining was used to detect alterations in the skin during the development of fetal skin and during the healing process of deep‑degree burned fetal skin. The expression levels of MMP‑9 and TIMP‑1 were determined using immunochemical staining, and their staining intensity was evaluated by mean optical density. The results demonstrated that fetal skin subcutaneously implanted into the dorsal skin flap of nude mice developed similarly to the normal growth process in the womb. In addition, the scarless healing process was clearly observed in the mice carrying the burned fetal skin. A total of 2 weeks was required to complete scarless healing. Following treatment with PBMCs, the burned fetal skin generated inflammatory factors and enhanced the inflammatory response, which consequently resulted in a reduction in the speed of healing and in the formation of scars. Therefore, exogenous PBMCs may alter the lowered immune response environment, which is required for scarless healing, resulting in scar formation. In conclusion, the present study indicated that the involvement of inflammatory cells is important during the healing process of deep‑degree burned skin, and MMP‑9 and TIMP‑1 may serve important roles in the process of scar formation.Keywords:
Nude mouse
Human skin
Skin repair
Epidermis (zoology)
Human skin
Skin Aging
Skin repair
Barrier function
Skin equivalent
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To determine changes in the morphology and function of pancreatic cancer cells after irreversible electroporation (IRE) treatment, and to explore the clinical significance of IRE treatment for pancreatic cancer providing an experimental basis for the clinical application of IRE treatment.IRE was carried out in an athymic nude mouse model of pancreatic carcinoma generated with human pancreatic cancer cells 1. In therapy groups, IRE electrodes were inserted with 90 pulses per second at 800 V/cm applied to ablate the targeted tumor tissues. Histological assessment of the affected tissue was performed by hematoxylin and eosin staining (HE). Quantification of cell proliferation and apoptosis was performed by evaluating Ki67 and caspase-3 levels, respectively. Flow cytometry was used to assess cell apoptosis. Ultrasound imaging was carried out to evaluate IRE treatment results. Pathological correlation studies showed IRE is effective for the targeted ablation of pancreatic tumors in an orthotopic mouse model.IRE was efficacious in removing tumors in the orthotopic mouse model. The IRE-ablated zone displays characteristics of nude mouse models at different time-points as assessed by hematoxylin and eosin staining. Immunohistochemical analysis of samples from the pancreatic cancer models showed significantly enhanced caspase-3 cleavage and Ki67. Flow cytometry data corroborated the above findings that apoptosis in tumor cells was observed immediately on the first postoperative day, and with time the middle and late stages of apoptosis were observed. For ultrasound imaging studies, the IRE ablation zone became a hyperechoic area due to increasing inflammatory and immunologic cellular contents.IRE is a promising new approach for pancreatic cancer, with many potential advantages over conventional ablation techniques.
Irreversible Electroporation
Nude mouse
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The skin is the largest organ of the human body and builds a barrier to protect us from the harmful environment and also from unregulated loss of water. Keratinocytes form the skin barrier by undergoing a highly complex differentiation process that involves changing their morphology and structural integrity, a process referred to as cornification. Alterations in the epidermal cornification process affect the formation of the skin barrier. Typically, this results in a disturbed barrier, which allows the entry of substances into the skin that are immunologically reactive. This contributes to and promotes inflammatory processes in the skin but also affects other organs. In many common skin diseases, including atopic dermatitis and psoriasis, a defect in the formation of the skin barrier is observed. In these diseases the cytokine composition within the skin is different compared to normal human skin. This is the result of resident skin cells that produce cytokines, but also because additional immune cells are recruited. Many of the cytokines found in defective skin are able to influence various processes of differentiation and cornification. Here we summarize the current knowledge on cytokines and their functions in healthy skin and their contributions to inflammatory skin diseases.
Human skin
Skin repair
Proinflammatory cytokine
Epidermis (zoology)
Barrier function
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Grafts of human skin on nude mice were irradiated with varying doses of ultraviolet light B and at various intervals were subjected to histological examination and determination of the epidermal <sup>3</sup>H-thymidine labelling index. The studies showed that the human skin in the foreign milieu of the nude mouse retained its ability to respond to phototoxic damage. The findings suggest that the nude mouse/human skin model could be a valuable tool in human photodermatological research.
Ultraviolet b
Morphology
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The aim of this study was to elaborate a histological model of incisional skin wound healing in Sprague-Dawley rats. Under aseptic conditions two paravertebral full thickness skin incisions were performed on the back of 42 anesthetized male rats. Histological sections from tissue specimens were stained by hematoxylin and eosin, van Gieson, PAS + PSD, Mallory's phosphotungstic hematoxylin and azur and eosin and evaluated during the first seven days after surgery. Histological evaluation revealed that the regeneration of injured epidermis was completed five days after surgery. The inflammatory phase was recorded during the first three days of healing with the culmination of this phase between day one and day two. The beginning of the proliferative phase was dated to the first day and the peak during day five and day six. The initiation of the maturation and remodeling phase of the healing process was observed six days after wounding. At the layer of striated muscle, the centronucleated cells were described for the first time six days after surgery. The wound healing process of rat skin was histologically described during the first seven days. Results of this work can serve as an experimental model for further research using external pharmacological and physical factors (laser light, magnetic field) by which the wound healing can be favourably influenced.
Van Gieson's stain
Skin repair
Histology
Granulation tissue
Eosin
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Artificial skin
Human skin
Skin repair
Skin equivalent
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Human skin
Epidermis (zoology)
Skin equivalent
Skin repair
Artificial skin
Ex vivo
Dermal fibroblast
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Abstract Human amniotic mesenchymal cells (hAMCs) and human amniotic epithelial cells (hAECs) have attracted increasing attention in recent years as a possible reserve of stem cells that may be useful for clinical application in regenerative medicine. The object of this study was to establish a new model for reconstruction of bilayered tissue‐engineered (TE) skin with hAMCs and hAECs (amniotic cells TE skin, AC‐TE skin). We studied these two types of cells and confirmed that they possessed the properties of stem cells. Mesenchymal–epidermal interactions are responsible for organogenesis. On the basis of this mechanism, we modified the constructing methods of traditional TE skin (TE skin with human fibroblasts and keratinocytes) and then established a new bilayered TE skin—AC‐TE skin. Histological and immunochemical methods were carried out to assess AC‐TE skin. The results showed that AC‐TE skin was similar in morphology to human skin which had stratified epidermis and underlying dermis. AC‐TE skin expressed proliferative cells marker Ki67 and epithelial stem cells marker K19; moreover, the constructed AC‐TE skin could successfully repair full thickness skin defects on athymic mice. Our findings suggest that AC‐TE skin is a useful skin equivalent which has good application prospects in regenerative medicine.
Human skin
Amniotic stem cells
Amniotic epithelial cells
Regenerative Medicine
Epidermis (zoology)
Skin repair
Amnion
Skin equivalent
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mechanobiology
Human skin
Skin equivalent
Ex vivo
Skin repair
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