Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells

Three-dimensional (3D) cell culture models are widely used to provide a more physiologically relevant microenvironment in which to host and study desired cell types. These models vary in complexity and cost, ranging from simple and inexpensive to highly sophisticated and costly systems. In this stud...

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Main Authors: Karla Pollyanna Vieira de Oliveira, Michael Yilma Yitayew, Ana Paula Almeida Bastos, Stefanie Cristine Nied Mandrik, Luismar Marques Porto, Maryam Tabrizian
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Journal of Functional Biomaterials
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Online Access:https://www.mdpi.com/2079-4983/16/6/208
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author Karla Pollyanna Vieira de Oliveira
Michael Yilma Yitayew
Ana Paula Almeida Bastos
Stefanie Cristine Nied Mandrik
Luismar Marques Porto
Maryam Tabrizian
author_facet Karla Pollyanna Vieira de Oliveira
Michael Yilma Yitayew
Ana Paula Almeida Bastos
Stefanie Cristine Nied Mandrik
Luismar Marques Porto
Maryam Tabrizian
author_sort Karla Pollyanna Vieira de Oliveira
collection DOAJ
description Three-dimensional (3D) cell culture models are widely used to provide a more physiologically relevant microenvironment in which to host and study desired cell types. These models vary in complexity and cost, ranging from simple and inexpensive to highly sophisticated and costly systems. In this study, we introduce a novel translucent multi-compartmentalized stacked multilayered nanocellulose scaffold and describe its fabrication, characterization, and potential application for co-culturing multiple cell types. The scaffold consists of bacterial nanocellulose (BNC) layers separated by interlayers of a lower density of nanocellulose fibers. Using this system, we co-cultured the MDA-MB-231 cell line with two tumor-associated cell types, namely BC-CAFs and M2 macrophages, to simulate the tumor microenvironment (TME). Cells remained viable and metabolically active for up to 15 days. Confocal microscopy showed no signs of cell invasion. However, BC-CAFs and MDA-MB-231 cells were frequently observed within the same layer. The expression of breast cancer-related genes was analyzed to assess the downstream functionality of the cells. We found that the E-cadherin expression was 20% lower in cancer cells co-cultured in the multi-compartmentalized scaffold than in those cultured in 2D plates. Since E-cadherin plays a critical role in preventing the initial dissociation of epithelial cells from the primary tumor mass and is often downregulated in the tumor microenvironment in vivo, this finding suggests that our scaffold more effectively recapitulates the complexity of a tumor microenvironment.
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spelling doaj-art-75e57c6c96944a7dbe7643f81dfa36b82025-08-20T03:24:39ZengMDPI AGJournal of Functional Biomaterials2079-49832025-06-0116620810.3390/jfb16060208Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing CellsKarla Pollyanna Vieira de Oliveira0Michael Yilma Yitayew1Ana Paula Almeida Bastos2Stefanie Cristine Nied Mandrik3Luismar Marques Porto4Maryam Tabrizian5Department of Chemical Engineering and Food Engineering, Technology Center, Federal University of Santa Catarina (UFSC), Campus Reitor João David Ferreira Lima, Florianópolis 88040-900, SC, BrazilBiomatX Lab, Department of Biomedical Engineering, Faculty of Medicine and Health Sciences, McGill University, 3775 University Street, Montreal, QC, H3A 2B6, CanadaEmbrapa Swine and Poultry, BR153, km 110, Tamanduá District, Concórdia 89715-899, SC, BrazilDepartment of Chemical Engineering and Food Engineering, Technology Center, Federal University of Santa Catarina (UFSC), Campus Reitor João David Ferreira Lima, Florianópolis 88040-900, SC, BrazilDepartment of Chemical Engineering and Food Engineering, Technology Center, Federal University of Santa Catarina (UFSC), Campus Reitor João David Ferreira Lima, Florianópolis 88040-900, SC, BrazilFaculty of Dental Medicine and Oral Health Sciences, McGill University, 2002 Avenue McGill College, Suite 500, Montreal, QC H2A 1G1, CanadaThree-dimensional (3D) cell culture models are widely used to provide a more physiologically relevant microenvironment in which to host and study desired cell types. These models vary in complexity and cost, ranging from simple and inexpensive to highly sophisticated and costly systems. In this study, we introduce a novel translucent multi-compartmentalized stacked multilayered nanocellulose scaffold and describe its fabrication, characterization, and potential application for co-culturing multiple cell types. The scaffold consists of bacterial nanocellulose (BNC) layers separated by interlayers of a lower density of nanocellulose fibers. Using this system, we co-cultured the MDA-MB-231 cell line with two tumor-associated cell types, namely BC-CAFs and M2 macrophages, to simulate the tumor microenvironment (TME). Cells remained viable and metabolically active for up to 15 days. Confocal microscopy showed no signs of cell invasion. However, BC-CAFs and MDA-MB-231 cells were frequently observed within the same layer. The expression of breast cancer-related genes was analyzed to assess the downstream functionality of the cells. We found that the E-cadherin expression was 20% lower in cancer cells co-cultured in the multi-compartmentalized scaffold than in those cultured in 2D plates. Since E-cadherin plays a critical role in preventing the initial dissociation of epithelial cells from the primary tumor mass and is often downregulated in the tumor microenvironment in vivo, this finding suggests that our scaffold more effectively recapitulates the complexity of a tumor microenvironment.https://www.mdpi.com/2079-4983/16/6/208bacterial nanocellulose3D cell culture modeltriple-cell co-culture
spellingShingle Karla Pollyanna Vieira de Oliveira
Michael Yilma Yitayew
Ana Paula Almeida Bastos
Stefanie Cristine Nied Mandrik
Luismar Marques Porto
Maryam Tabrizian
Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells
Journal of Functional Biomaterials
bacterial nanocellulose
3D cell culture model
triple-cell co-culture
title Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells
title_full Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells
title_fullStr Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells
title_full_unstemmed Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells
title_short Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells
title_sort transparent 3 layered bacterial nanocellulose as a multicompartment and biomimetic scaffold for co culturing cells
topic bacterial nanocellulose
3D cell culture model
triple-cell co-culture
url https://www.mdpi.com/2079-4983/16/6/208
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