resumo
A gene-activated surface coating is presented as a strategy to design smart biomaterials for bone tissue engineering. The thin-film coating is based on polyelectrolyte multilayers composed of collagen I and chondroitin sulfate, two main biopolymers of the bone extracellular matrix, which are fabricated by layer-by-layer assembly. For further functionalization, DNA/lipid-nanoparticles (lipoplexes) are incorporated into the multilayers. The polyelectrolyte multilayer fabrication and lipoplex deposition are analyzed by surface sensitive analytical methods that demonstrate successful thin-film formation, fibrillar structuring of collagen, and homogenous embedding of lipoplexes. Culture of mesenchymal stem cells on the lipoplex functionalized multilayer results in excellent attachment and growth of them, and also, their ability to take up cargo like fluorescence-labelled DNA from lipoplexes. The functionalization of the multilayer with lipoplexes encapsulating DNA encoding for transient expression of bone morphogenetic protein 2 induces osteogenic differentiation of mesenchymal stem cells, which is shown by mRNA quantification for osteogenic genes and histochemical staining. In summary, the novel gene-functionalized and extracellular matrix mimicking multilayer composed of collagen I, chondroitin sulfate, and lipoplexes, represents a smart surface functionalization that holds great promise for tissue engineering constructs and implant coatings to promote regeneration of bone and other tissues.
palavras-chave
POLYELECTROLYTE MULTILAYER FILMS; BONE MORPHOGENETIC PROTEINS; CHONDROITIN SULFATE; SUBSTRATE STIFFNESS; CATIONIC LIPIDS; COLLAGEN I; TISSUE; DNA; BMP-2; TRANSFECTION
categoria
Engineering; Science & Technology - Other Topics; Materials Science
autores
Husteden, C; Barrera, YAB; Tegtmeyer, S; Borges, J; Giselbrecht, J; Menzel, M; Langner, A; Mano, JF; Schmelzer, CEH; Wölk, C; Groth, T
nossos autores
João Borges
Investigador Auxiliar
João Mano
Vice-DiretorProjectos
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)
agradecimentos
C.H. and Y.A.B.B. contributed equally to this work. The authors are thankful for the help and support of Dr. Navarrete Santos during flow cytometry measurements and data analysis. Y.A.B.B was supported by the Consejo Nacional de Ciencia y Tecnologia (CONACYT-Mexico) and Deutscher Akademischer Austauschdienst (DAAD) for funding. Furthermore, she received a grant from the International Graduate School AGRIPOLY supported by the European Regional Development Fund (ERDF) and Ministerium fuer Wissenschaft und Wirtschaft, Land Sachsen-Anhalt. C.H. was supported by DAAD which funded the exchange to University Aveiro. This work was funded by the Fraunhofer Internal Programs under Grant No. Attract 069-608203 (C.E.H.S.) and by the Programa Operacional Regional do Centro - Centro 2020, in the component FEDER, and by national funds (OE) through Fundacao para a Ciencia e a Tecnologia/Ministerio da Ciencia, Tecnologia e Ensino Superior (FCT/MCTES), in the scope of the project "SUPRASORT" (PTDC/QUI-OUT/30658/2017, CENTRO-01-0145-FEDER-030658). This work was developed within the scope of the project CICECO - Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020, financed by national funds through the FCT/MEC (PIDDAC). The support by the Deutsche Forschungsgemeinschaft (DFG) project-ID 396823779 (C.W.) is acknowledged. J.B. gratefully acknowledges FCT for the individual Assistant Researcher contract (2020.00758.CEECIND). Open Access funding enabled and organized by Projekt DEAL.