A modified hydrogel production protocol to decrease cellular content.

IF 1.1 4区医学 Q3 SURGERY Acta cirurgica brasileira Pub Date : 2022-01-01 DOI:10.1590/acb371005

Gabriela Catão Diniz Braga, Cristina Pires Camargo, Martin Conrad Harmsen, Aristides Tadeu Correia, Sonia Souza, Marilia Seelaender, Viviane Araujo Nunes, Jeniffer Farias Dos Santos, Elida Adalgisa Neri, Iuri Cordeiro Valadão, Luiz Felipe Pinho Moreira, Rolf Gemperli

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Abstract

Purpose: To analyze the cytotoxicity and cell in porcine-derived decellularized skin matrix.

Methods: We analyzed the effect of multiple decellularization processes by histological analysis, DNA quantification, and flow cytometry. Subsequently, we analyzed the most appropriate hydrogel concentration to minimize cytotoxicity on fibroblast culture and to maximize cell proliferation.

Results: After the fourth decellularization, the DNA quantification showed the lowest DNA concentration (< 50 ng/mg). Histological analysis showed no cell components in the hydrogel. Moreover, hematoxylin and eosin showed a heterogeneous structure of collagen fibers. The best hydrogel concentration ranged from 3 to 25%, and there was no significant difference between the 24 hours and seven days.

Conclusions: The process of hydrogel production was effective for removing cells and DNA elements. The best hydrogel concentration ranged from 3 to 25%.

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一种改进的水凝胶生产方案，以减少细胞含量。

目的:分析猪源脱细胞皮肤基质的细胞毒性和细胞活性。方法:采用组织学分析、DNA定量和流式细胞术分析多种脱细胞过程的影响。随后，我们分析了最合适的水凝胶浓度，以减少成纤维细胞培养的细胞毒性，并最大化细胞增殖。结果:第四次脱细胞后，DNA定量显示DNA浓度最低(< 50 ng/mg)。组织学分析显示水凝胶中没有细胞成分。苏木精和伊红的胶原纤维结构呈异质结构。最佳水凝胶浓度为3% ~ 25%，24 h与7 d间无显著差异。结论:水凝胶的制备工艺能有效去除细胞和DNA元素。最佳水凝胶浓度为3% ~ 25%。

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