Application of hydrogel wound dressings in cell therapy-approaches to assessment in vitro.

IF 1.4 Q3 EMERGENCY MEDICINE International Journal of Burns and Trauma Pub Date : 2023-01-01
Marfa N Egorikhina, Irina I Kobyakova, Irina N Charykova, Daria D Linkova, Yulia P Rubtsova, Ekaterina A Farafontova, Diana Ya Aleynik
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Abstract

Cell therapy is actively used to treat skin defects, particularly burn lesions. The effectiveness of its application may depend on the appropriate choice of wound dressings used together with any cellular material. The aim of the study was to investigate the interaction of 4 hydrogel dressings used in clinical practice with human cells in an in vitro model to determine if their use in combination with cell therapy is possible. The effect of the dressings on the growth medium was assessed by considering the changes caused in the medium's acid-base equilibrium (pH) and viscosity. Cytotoxicity was determined by applying an MTT-assay and by direct contact methods. Cell adhesion and viability on the dressing surfaces were analyzed using fluorescence microscopy. Proliferative and secretory cell activity were determined concurrently. Characterized human dermal fibroblast cultures were used as the test cultures. Results: The tested dressings interacted differently with the growth medium and the test cultures. 1-day extracts of all dressings had almost no effect on the acid-base balance, but, after 7 days, the pH of the dressing Type 2 extract had sharply acidified. The viscosity of the media under the influence of dressings of Types 2 and 3 had also markedly increased. MTT-assays showed nontoxicity of all the 1-day-incubated dressing extracts, while incubation for 7-days resulted in extracts with evident cytotoxicity, which was reduced upon dilution. Cell adhesion to the surfaces of the dressings differed, being observed occurring on dressings 2 and 3, and to a limited extent on dressing 4. Cells under dressing 1 showed evident proliferative and secretory activity whereas the other dressings impaired either proliferation or secretion processes. These effects indicate that, in general, comprehensive studies including a variety of methodological approaches at the in vitro stage are needed to allow the selection of appropriate dressings if they are to be used in combination with cell therapy to act as cell carriers. Of those investigated, the Type 1 dressing can be recommended as a protective dressing for use after transplantation of cells into a wound defect area by some other method.

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水凝胶伤口敷料在细胞治疗中的应用——体外评估方法。
细胞疗法被积极用于治疗皮肤缺陷,特别是烧伤损伤。其应用的有效性可能取决于与任何细胞材料一起使用的伤口敷料的适当选择。该研究的目的是在体外模型中研究临床实践中使用的4种水凝胶敷料与人体细胞的相互作用,以确定它们是否可能与细胞疗法联合使用。通过考虑培养基的酸碱平衡(pH)和粘度的变化来评估敷料对生长培养基的影响。采用mtt法和直接接触法测定细胞毒性。荧光显微镜观察细胞在敷料表面的粘附和活力。同时测定增殖和分泌细胞的活性。以人真皮成纤维细胞培养物为试验培养物。结果:实验敷料与培养基和实验培养物的相互作用不同。第1天,所有敷料的提取物对酸碱平衡几乎没有影响,但7天后,敷料2型提取物的pH急剧酸化。在2、3型敷料的影响下,介质的粘度也明显增加。mtt试验显示,所有培养1天的敷料提取物无毒,而培养7天的提取物有明显的细胞毒性,稀释后降低。细胞对敷料表面的粘附不同,在敷料2和3上观察到,并且在有限程度上在敷料4上出现。敷料1下的细胞表现出明显的增殖和分泌活性,而其他敷料对细胞的增殖和分泌过程均有损害。这些影响表明,一般来说,如果要将敷料与细胞疗法结合使用,作为细胞载体,需要在体外阶段进行包括各种方法方法在内的综合研究,以便选择合适的敷料。在这些研究中,1型敷料可以推荐作为一种保护性敷料,用于通过其他方法将细胞移植到伤口缺损区域后使用。
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