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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

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

作者信息

Egorikhina Marfa N, Kobyakova Irina I, Charykova Irina N, Linkova Daria D, Rubtsova Yulia P, Farafontova Ekaterina A, Aleynik Diana Ya

机构信息

Federal State Budgetary Educational Institution of Higher Education, Privolzhsky Research Medical University of The Ministry of Health of The Russian Federation Nizhny Novgorod 603005, Russia.

出版信息

Int J Burns Trauma. 2023 Apr 15;13(2):13-32. eCollection 2023.

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.

摘要

细胞疗法正被积极用于治疗皮肤缺损,尤其是烧伤创面。其应用效果可能取决于与任何细胞材料一起使用的伤口敷料的恰当选择。本研究的目的是在体外模型中研究临床实践中使用的4种水凝胶敷料与人类细胞的相互作用,以确定它们与细胞疗法联合使用是否可行。通过考虑敷料对培养基酸碱平衡(pH值)和粘度的影响来评估敷料对生长培养基的作用。通过MTT法和直接接触法测定细胞毒性。使用荧光显微镜分析敷料表面的细胞粘附和活力。同时测定细胞的增殖和分泌活性。使用特征明确的人真皮成纤维细胞培养物作为测试培养物。结果:测试的敷料与生长培养基和测试培养物的相互作用不同。所有敷料的1天提取物对酸碱平衡几乎没有影响,但7天后,2型敷料提取物的pH值急剧酸化。2型和3型敷料作用下培养基的粘度也显著增加。MTT分析表明,所有1天培养的敷料提取物均无毒性,而培养7天则导致提取物具有明显的细胞毒性,稀释后毒性降低。细胞在敷料表面的粘附情况不同,在2型和3型敷料上观察到细胞粘附,在4型敷料上细胞粘附程度有限。1型敷料下的细胞表现出明显的增殖和分泌活性,而其他敷料则损害了增殖或分泌过程。这些结果表明,一般来说,如果要将敷料与细胞疗法联合使用作为细胞载体,在体外阶段需要进行包括多种方法的综合研究,以便选择合适的敷料。在所研究的敷料中,1型敷料可推荐为在通过其他方法将细胞移植到伤口缺损区域后使用的保护性敷料。

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