Evaluation of Polycaprolactone Electrospun Nanofiber-Composites for Artificial Skin Based on Dermal Fibroblast Culture

The study’s aim was to develop a dermal equivalent scaffold that can mimic the architecture and biological performance of the human dermis. Poly ε-caprolactone (PCL) electrospun nanofiber material (ENF) was assembled with polyethylene glycol diacrylate (PEGDA), sodium alginate (SA) and type I collagen (CG1) to develop three groups of dermal equivalent scaffolds. These scaffolds were named PEGDA-PCL, SA-PCL and CG1-PCL. Scanning electron microscopy (SEM) images of cell-free scaffolds’ top and cross-sectional surface were collected and analyzed to examine internal morphology, specifically the adhesiveness of PCL fibers with the different scaffolds. Human dermal fibroblasts were cultured on each of the scaffolds. Cell viability studies including cell adhesion, cell differentiation and stress fiber production were conducted on each scaffold. Furthermore, the architectural integrity of each scaffold was verified by degradation analysis for 2 weeks by soaking each scaffold in phosphate-buffered saline (PBS) solution. Finally, we conducted rheological characteristics of each scaffold. Based on our results from the above analysis, the study concluded that CG1-PCL is best suitable for the dermal equivalent model and has potential to be used as a graft for skin repair.

Tissue dermal equivalent – а cellular product based on human dermal keratinocytes and fibroblasts: the properties of equivalent components and perspectives of practical application

The nature and ways of isolation and cultivation in vitro of keratinocytes and fibroblasts, the main cellular components of skin to prepare a new biomedical product, tissue dermal equivalent were considered. The main attention was payed to optimization of upbuilding dermal cell biomass including selection of medium compositions and conditions of cultivation. The information was given on main parameters of cell cultures as proliferation activity, viability and phenotype of the cells. Genotoxicity of fibroblasts and biocompatibility of the cells with organic matrixes to find the optimal carrier for cellular elements of tissue dermal equivalent were studied. The composition, the process of preparation of tissue dermal equivalent and perspectives of its practical application were discussed.

A Novel In Vitro Culture Model System to Study Merkel Cell Polyomavirus–Associated MCC Using Three-Dimensional Organotypic Raft Equivalents of Human Skin

Merkel cell polyomavirus (MCPyV) is a human polyomavirus causally linked to the development of Merkel cell carcinoma (MCC), an aggressive malignancy that largely arises within the dermis of the skin. In this study, we recapitulate the histopathology of human MCC tumors in vitro using an organotypic (raft) culture system that is traditionally used to recapitulate the dermal and epidermal equivalents of skin in three dimensions (3D). In the optimal culture condition, MCPyV+ MCC cells were embedded in collagen between the epidermal equivalent comprising human keratinocytes and a dermal equivalent containing fibroblasts, resulting in MCC-like lesions arising within the dermal equivalent. The presence and organization of MCC cells within these dermal lesions were characterized through biomarker analyses. Interestingly, co-culture of MCPyV+ MCC together with keratinocytes specifically within the epidermal equivalent of the raft did not reproduce human MCC morphology, nor were any keratinocytes necessary for MCC-like lesions to develop in the dermal equivalent. This 3D tissue culture system provides a novel in vitro platform for studying the role of MCPyV T antigens in MCC oncogenesis, identifying additional factors involved in this process, and for screening potential MCPyV+ MCC therapeutic strategies.

Morphological characteristics of experimental ischemic wound healing after application of autoand hetero-fibroblasts and dermal equivalent

Цель. Изучить морфологическое строение, коллагенообразование и ангиогенез в биоптатах новообразованного эпидермиса и дермы на 19-е сутки их восстановления в экспериментальной ишемизированной ране после введения ауто- и гетерофибробластов, а также после трансплантации дермального эквивалента с гетерофибробластами. Методы. Исследование выполнено на 28 белых половозрелых мышах линии С57/В1 в возрасте до 1 года. Вокруг и в дно хирургической экспериментальной кожной раны в лопаточной области вводили 0,4 мл взвеси фибробластов и дермальный эквивалент в количестве 1,33 млн клеток. Биоптат заливали в парафин, окрашивали гематоксилином и эозином и по Вейгерту-Ван-Гизону. Результаты. Наиболее существенно на регенераторный гистогенез влияет введение аутофибробластов, когда фиксируется наибольшая толщина эпидермиса, наиболее активны ангиогенез и коллагенообразование. Вместе с тем, самым дифференцированным эпидермис становится после трансплантации в рану дермального эквивалента с гетерофибробластами. Вывод. Благоприятное воздействие дермального эквивалента с гетерофибробластами отличается от воздействия взвеси аутофибробластов всего на несколько процентов: толщина эпидермиса - на 4,29%, площадь коллагеновых волокон - на 2,66%, площадь кровеносных сосудов - на 4,04%, что делает такие различия недостоверными.