Doxycycline-doped membranes induced osteogenic gene expression on osteoblastic cells

Manuel Toledano-Osorio; Francisco Javier Manzano-Moreno; Manuel Toledano; Raquel Osorio; Antonio L Medina-Castillo; Víctor J Costela-Ruiz; Concepción Ruiz

doi:10.1016/j.jdent.2021.103676

Doxycycline-doped membranes induced osteogenic gene expression on osteoblastic cells

J Dent. 2021 Jun:109:103676. doi: 10.1016/j.jdent.2021.103676. Epub 2021 Apr 20.

Authors

Manuel Toledano-Osorio¹, Francisco Javier Manzano-Moreno², Manuel Toledano³, Raquel Osorio⁴, Antonio L Medina-Castillo⁵, Víctor J Costela-Ruiz⁶, Concepción Ruiz⁷

Affiliations

¹ Biomaterials in Dentistry Research Group, Department of Stomatology, School of Dentistry, University of Granada, Spain; Medicina Clínica y Salud Pública PhD Programme, Spain.
² Biomedical Group (BIO277), Department of Stomatology, School of Dentistry, University of Granada, Spain; Instituto Investigación Biosanitaria, ibs, Granada, Granada, Spain.
³ Biomaterials in Dentistry Research Group, Department of Stomatology, School of Dentistry, University of Granada, Spain. Electronic address: toledano@ugr.es.
⁴ Biomaterials in Dentistry Research Group, Department of Stomatology, School of Dentistry, University of Granada, Spain.
⁵ NanoMyP®, Nanomateriales y Polimeros S.L., Spin-Off company of the University of Granada, Edificio BIC-Granada. Av. Innovación 1. 18016 - Armilla, Granada, Spain.
⁶ Instituto Investigación Biosanitaria, ibs, Granada, Granada, Spain; Biomedical Group (BIO277), Department of Nursing, Faculty of Health Sciences, University of Granada, Spain.
⁷ Instituto Investigación Biosanitaria, ibs, Granada, Granada, Spain; Biomedical Group (BIO277), Department of Nursing, Faculty of Health Sciences, University of Granada, Spain; Institute of Neuroscience, University of Granada, Centro de Investigación Biomédica (CIBM), Parque de Tecnológico de la Salud (PTS), Granada, Spain.

PMID: 33891990
DOI: 10.1016/j.jdent.2021.103676

Abstract

Objectives: To analyze how novel developed silicon dioxide composite membranes, functionalized with zinc or doxycycline, can modulate the expression of genes related to the osteogenic functional capacity of osteoblastic cells.

Methods: The composite nanofibers membranes were manufactured by using a novel polymeric blend and 20 nm silicon dioxide nanoparticles (SiO₂-NPs). To manufacture the membranes, 20 nm SiO₂-NPs were added to the polymer solution and the resulting suspension was processed by electrospinning. In a second step, the membranes were functionalized with zinc or doxycycline. Then, they were subjected to MG63 osteoblast-like cells culturing for 48 h. After this time, real-time quantitative polymerase chain reaction (RT-qPCR) was carried out to study the expression of Runx-2, OSX, ALP, OSC, OPG, RANKL, Col-I, BMP-2, BMP-7, TGF-β1, VEGF, TGF-βR1, TGF- βR2, and TGF-βR3. Mean comparisons were conducted by One-way ANOVA and Tukey tests (p < 0.05).

Results: In general, the blending of SiO₂-NPs in the tested non-resorbable polymeric scaffold improves the expression of osteogenic genes over the control membranes. Doxycycline doping of experimental scaffolds attained the best results, encountering up-regulation of BMP-2, ALP, OPG, TGFβ-1 and TGFβ-R1. Membranes with zinc induced a significant increase in the expression of Col-I, ALP and TGF β1. Both, zinc and doxycycline functionalized membranes enormously down-regulated the expression of RANKL.

Conclusions: Zinc and doxycycline doped membranes are bioactive inducing overexpression of several osteogenic gene markers.

Clinical significance: Doxycycline doped membranes may be a potential candidate for use in GBR procedures in several challenging pathologies, including periodontal diseases.

Keywords: Doxycycline; GBR; Gene expression; Membrane; RT-qPCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Differentiation
Doxycycline*
Gene Expression
Osteoblasts
Osteogenesis
Silicon Dioxide*

Substances

Silicon Dioxide
Doxycycline