Co-cultivation of Komagataeibacter sp. and Lacticaseibacillus sp. strains to produce bacterial nanocellulose-hyaluronic acid nanocomposite membranes for skin wound healing applications

abstract

Inspired by natural microbial cooperation, a co-culture approach was used to synthesize bacterial nanocellulose (BNC)-based nanocomposites for potential wound healing applications. By co-culturing either Komagataeibacter xylinus (K1G4) or the never tested strain K. rhaeticus (K2G46) with the hyaluronic acid (HA)-producer Lacticaseibacillus casei UMCC 2535, two BNC-HA nanocomposites were obtained (C1-K1 and C2-K2). The membranes showed a HA content of 0.49 f 0.05 mg (C1-K1) and 1.40 f 0.07 mg (C2-K2), and both revealing a nearly complete release of HA after 1 h in PBS. Compared to pure BNC membranes, the nanocomposites showed enhanced properties, including higher crystallinity (K1G4 = 84.6 %; K2G46 = 76.5 %; C1-K1 = 89.1 %; C2-K2 = 88.1 %), and Young's modulus (K1G4 = 3.38 f 0.56 GPa; K2G46 = 2.22 f 0.65 GPa; C1-K1 = 10.00 f 1.32 GPa; C2-K2 = 7.90 f 1.54 GPa). Additionally, both BNC-HA membranes exhibited increased moisture uptake (K1G4 = 9.06 f 0.47 %; K2G46 = 9.27 f 1.33 %; C1-K1 = 13.65 f 0.53 %; C2-K2 = 16.26 f 1.05 %) and water absorption (K1G4 = 82.18 f 5.25 %; K2G46 = 86.54 f 7.86 %; C1-K1 = 160.04 f 9.33 %; C2-K2 = 144.42 f 13.86 %) capacity. Moreover, they were non-cytotoxic towards human keratinocyte (HaCaT) cells, with >90 % cell viability for up to 72 h. The in vitro scratch assays showed a complete wound closure within 48 h for cells exposed to BNC-HA membranes. These findings underscore the potential of co-culturing system to develop BNCHA nanocomposites for wound healing applications.

keywords

IN-SITU; CARBON-SOURCES; CELLULOSE; FILMS; CHITOSAN

subject category

Biochemistry & Molecular Biology; Chemistry; Polymer Science

authors

Brugnoli, M; Carvalho, JPF; Arena, MP; Oliveira, H; Vilela, C; Freire, CSR; Gullo, M

Groups

G4 - Renewable Materials and Circular Economy

Projects

Collaboratory for Emerging Technologies, CoLab (EMERGING TECHNOLOGIES)

CICECO - Aveiro Institute of Materials (UIDB/50011/2020)

CICECO - Aveiro Institute of Materials (UIDP/50011/2020)

Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)

acknowledgements

Part of this work was supported by the European Commission - NextGenerationEU, Project SUS-MIRRI.IT "Strengthening the MIRRI Italian Research Infrastructure for Sustainable Bioscience and Bio-economy", code n. IR0000005, and by the European Union - NextGe-nerationEUGrant, CN_00000033, Project "National Biodiversity Future Center - NBFC". CUP E93C22001090001. This work was also developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 (DOI 10.54499/UIDB/50011/2020) , UIDP/50011/2020 (DOI 10.54499/UIDP/50011/2020) & LA/P/0006/2020 (DOI 10.54499/LA/P/0006/2020) , financed by national funds through the FCT/MEC (PIDDAC) . FCT is acknowledged for the doctoral grants to J.P. F.C (2020.09018.BD, DOI 10.54499/2020.09018.BD) and the research contracts under Scientific Employment Stimulus to C.S.R.F. (DOI 10. 54499/CEE CIND/00464/2017/CP1459/CT0033) , C. V . (DOI 10. 54499/2021.01571.CEECIND/CP16 59/CT0024) and H.O. (DOI 10 .54499/CEECIND/04050/2017/CP1459/CT0023) .