Abstract

Bacterial cellulose (BC)–gelatin (GEL) membranes were processed by successive periodate oxidation and a freeze-thawing/carbodiimide crosslinking procedure, first facilitating a Schiff-base reaction among respective aldehyde and hydroxyl groups, and later GEL stabilization and microstructuring. The formation of highly microporous structures within the GEL portion, with significant differences between bottom and top, was elucidated, and pores in the 27.6 ± 3 µm–108 ± 5 µm range were generated, exceeding the threshold value of ~10 µm sufficient for cell trafficking. During a relatively short (6 h) exhaustion procedure in supersaturated simulated body fluid solution, the membranes accommodated the combination of biologically relevant minerals, i.e., flake-like octacalcium phosphate (OCP) and (amorphous) apatite, onto their surface, forming a membrane with intensive swelling (650–1650%) and up to 90% weight loss in a 4-week period. The membranes´ 6-day eluates did not evoke any cytotoxic effects toward human fibroblast, MRC-5 cells. The same type of cells retained their morphology in direct contact with the membrane, attaching to the GEL porous site, while not attaching to the GEL thin-coated BC side, most probably due to combined, ablation effect of dominant β-sheet conformation and carbodiimide crosslinking. Together with arrested proliferation through the BC side, the membranes demonstrated beneficial properties for potential guided tissue regeneration (GTR) applications.

Highlights

  • Bacterial cellulose (BC) is a nanofibrilar, firmly structured polysaccharide, produced extracellularly by the gram-negative, acetic acid bacteria Komagataeibacter xylinus

  • We hypothesized the following: 1. A BC membrane can be efficiently merged with porous GEL through a simple procedure we propose, using a water-based system free of including additional chemicals; 2

  • The periodate-mediated oxidation of BC is reported to proceed through redox cleavage of vicinal (C2−C3) glycols, which yields a product with aldehyde groups in the positions C2 and C3 of the glucopiranose unit, i.e., 2,3-dialdehyde BC [21]

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Summary

Introduction

Bacterial cellulose (BC) is a nanofibrilar, firmly structured polysaccharide, produced extracellularly by the gram-negative, acetic acid bacteria Komagataeibacter xylinus (the previous Gluconcetobacter xylinus [1]) This nanomaterial, consisting of randomly assembled,

Reagents and Materials
BC–GEL Membranes
In Situ Mineralization
Swelling and Degradation Test
Cell Line
Preparation of Eluates and Cellular Exposure
Direct Exposure of Cells with Samples
Bioreduction into Formazan
Analysis in Direct Contact
BC Modification
Microstructural Assessment
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