Shape-Versatile Fixed Cellular Materials for Multiple Target Immunomodulation

abstract

Therapeutic cells are usually administered as living agents, despite the risks of undesired cell migration and acquisition of unpredictable phenotypes. Additionally, most cell-based therapies rely on the administration of single cells, often associated with rapid in vivo clearance. 3D cellular materials may be useful to prolong the effect of cellular therapies and offer the possibility of creating structural volumetric constructs. Here, the manufacturing of shape-versatile fixed cell-based materials with immunomodulatory properties is reported. Living cell aggregates with different shapes (spheres and centimeter-long fibers) are fixed using a method compatible with maintenance of structural integrity, robustness, and flexibility of 3D constructs. The biological properties of living cells can be modulated before fixation, rendering an in vitro anti-inflammatory effect toward human macrophages, in line with a decreased activation of the nuclear factor kappa B (NF-kappa B) pathway that preponderantly correlated with the surface area of the materials. These findings are further corroborated in vivo in mouse skin wounds. Contact with fixed materials also reduces the proliferation of activated primary T lymphocytes, while promoting regulatory populations. The fixation of cellular constructs is proposed as a versatile phenotypic stabilization method that can be easily implemented to prepare immunomodulatory materials with therapeutic potential. Aggregates of cells undergo a fixation procedure to ensure devitalization and long-lasting maintenance of architectonic features, both for spheroids and fibers. Direct contact of fixed cellular materials with cells from the innate and adaptive branches of the immune system shows their immunomodulatory properties. In an in vivo wound model distinct immunomodulatory profiles are dependent on the administered dose. image

keywords

MESENCHYMAL STEM-CELLS; TISSUE; TRANSPLANTATION

subject category

Chemistry; Science & Technology - Other Topics; Materials Science; Physics

authors

Sousa, AR; Cunha, AF; Santos-Coquillat, A; Estrada, BH; Spiller, KL; Barao, M; Rodrigues, AF; Simoes, S; Vilaca, A; Ferreira, L; Oliveira, MB; Mano, JF

Groups

G5 - Biomimetic, Biological and Living Materials

Projects

Advancing the Regenerative and Translational Potential of Cellular Fibers (CellFi)

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)

Collaboratory for Emerging Technologies, CoLab (EMERGING TECHNOLOGIES)

acknowledgements

A.R.S. and A.F.C. contributed equally to this work. This work was supported by the Programa Operacional Competitividade e Internacionalizac & atilde;o, in the component FEDER, and by national funds (OE) through FCT/MCTES, in the scope of the project "CellFi", PTDC/BTM-ORG/3215/2020 (DOI10.54499/PTDC/BTM-ORG/3215/2020); "Prova de conceito" project with the acronym "Bio-Med" (Ref. 181228) funded by COMPETE2020 and PRR project "Agenda/Aliancas mobilizadoras para a reindustrilizac & atilde;o" (Ref. C630926586-00465198); and PRR through the project HfPT-Health from Portugal (Ref. 02/C05-i01.01/2022.PC644937233-00000047). This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 (DOI10.54499/UIDB/50011/2020; DOI10.54499/UIDP/50011/2020), and LA/P/0006/2020 (DOI10.54499/LA/P/0006/2020), financed by national funds through the FCT/MCTES (PIDDAC). A.R.S. acknowledges the Ph.D. grant (SFRH/BD/145765/2019) and the FLAD grant (Project No. 2023/0222). A.F.C. acknowledges the Ph.D. grant 2021.04817 (DOI10.54499/2021.04817.BD). M.B.O. acknowledges national funds through FCT - Fundac & atilde;o para a Ciencia e a Tecnologia, I.P., under the Scientific Employment Stimulus - Institutional Call - CEECINST/00013/2021 (DOI10.54499/CEECINST/00013/2021/CP2779/CT0010). K.L.S acknowledges NHLBI grant R01 HL130037. This project utilized the Flow Cytometry and Human Immune Monitoring Shared Resources at the Sidney Kimmel Cancer Center, supported by the NCI grant (5P30CA056036-17).