Nanostructured Cellulose–Gellan–Xyloglucan–Lysozyme Dressing Seeded with Mesenchymal Stem Cells for Deep Second-Degree Burn Treatment

Carolina Maria Costa de Oliveira Souza,¹ Clayton Fernandes de Souza,^2,3 Bassam Felipe Mogharbel,¹ Ana Carolina Irioda,¹ Celia Regina Cavichiolo Franco,⁴ Maria Rita Sierakowski,³ Katherine Athayde Teixeira de Carvalho<sup>1

1</sup>Stem Cell Research Laboratory, Cell Therapy and Biotechnology in Regenerative Medicine Department, Pequeno Príncipe Faculties and the Pelé Pequeno Príncipe Research Institute, Curitiba, Paraná, Brazil; ²Chemistry Undergraduate Program, School of Education and Humanities, Pontifícia Universidade Católica Do Paraná, Curitiba, Paraná, Brazil; ³BioPol, Chemistry Department, Federal University of Paraná, Curitiba, Paraná, Brazil; ⁴Molecular Biology Department, Federal University of Paraná, Curitiba, Paraná, Brazil

Correspondence: Katherine Athayde Teixeira de Carvalho
Stem Cell Research Laboratory, Cell Therapy and Biotechnology in Regenerative Medicine Department, Pequeno Príncipe Faculties and the Pelé Pequeno Príncipe Research Institute, 1632 Avenida Silva Jardim, Box 80.240-020, Curitiba, Paraná, Brazil
Tel +55 41-3310-1034
Email katherinecarv@gmail.com

Purpose: In deep burns, wound contraction and hypertrophic scar formation can generate functional derangement and debilitation of the affected part. In order to improve the quality of healing in deep second-degree burns, we developed a new treatment in a preclinical model using nanostructured membranes seeded with mesenchymal stem cells (MSCs).
Methods: Membranes were obtained by reconstitution of bacterial cellulose (reconstituted membrane [RM]) and produced by a dry-cast process, then RM was incorporated with 10% tamarind xyloglucan plus gellan gum 1:1 and 10% lysozyme (RMGT–LZ) and with 10% gellan gum and 10% lysozyme (RMG–LZ). Membrane hydrophobic/hydrophilic characteristics were investigated by static/dynamic contact-angle measurements. They were cultivated with MSCs, and cell adhesion, proliferation, and migration capacity was analyzed with MTT assays. Morphological and topographic characteristics were analyzed by scanning electron microscopy. MSC patterns in flow cytometry and differentiation into adipocytes and osteocytes were checked. In vivo assays used RMG–LZ and RMGT–LZ (with and without MSCs) in Rattus norvegicus rats submitted to burn protocol, and histological sections and collagen deposits were analyzed and immunocytochemistry assay performed.
Results: In vitro results demonstrated carboxyl and amine groups made the membranes moderately hydrophobic and xyloglucan inclusion decreased wettability, favoring MSC adhesion, proliferation, and differentiation. In vivo, we obtained 40% and 60% reduction in acute/chronic inflammatory infiltrates, 96% decrease in injury area, increased vascular proliferation and collagen deposition, and complete epithelialization after 30 days. MSCs were detected in burned tissue, confirming they had homed and proliferated in vivo.
Conclusion: Nanostructured cellulose–gellan–xyloglucan–lysozyme dressings, especially when seeded with MSCs, improved deep second-degree burn regeneration.

Keywords: skin, cell-based therapy, transplantation, hydrocolloid dressing