Doxycycline-doped collagen membranes accelerate in vitro osteoblast proliferation and differentiation Toledano Osorio, Manuel Luna Bertos, María Elvira De Toledano Pérez, Manuel Manzano Moreno, Francisco Javier García Recio, Enrique Ruiz Rodríguez, Concepción Osorio Ruiz, Raquel Sanz, Mariano bone regeneration dexamethasone doxycycline membranes Osteoblasts Objective: The aim of the study was to evaluate the effect of doxycycline- and dexamethasone-doped collagen membranes on the proliferation and differentiation of osteoblasts. Background: Collagen barrier membranes are frequently used to promote bone regeneration and to boost this biological activity their functionalization with antibacterial and immunomodulatory substances has been suggested. Methods: The design included commercially available collagen membranes doped with doxycycline (Dox-Col-M) or dexamethasone (Dex-Col-M), as well as undoped membranes (Col-M) as controls, which were placed in contact with cultured MG63 osteoblast-like cells (ATCC). Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay and differentiation by measuring the alkaline phosphatase (ALP) activity using spectrophotometry. Real-time quantitative polymerase chain reaction was used to study the expression of the genes: Runx-2, OSX, ALP, OSC, OPG, RANKL, Col-I, BMP-2, BMP-7, TGF-β1, VEGF, TGF-βR1, TGF-βR2, and TGF-βR3. Scanning electron microscopy was used to study osteoblast morphology. Data were assessed using one-way analysis of variance or Kruskal-Wallis tests, once their distribution normality was assessed by Kolmogorov-Smirnov tests (p > .05). Bonferroni for multiple comparisons were carried out (p < .05). Results: Osteoblast proliferation was significantly enhanced in the functionalized membranes as follows: (Col-M < Dex-Col-M < Dox-Col-M). ALP activity was significantly higher on cultured osteoblasts on Dox-Col-M. Runx-2, OSX, ALP, OSC, BMP-2, BMP-7, TGF-β1, VEGF, TGF-βR1, TGF-βR2, and TGF-βR3 were overexpressed, and RANKL was down-regulated in osteoblasts cultured on Dox-Col-M. The osteoblasts cultured in contact with the functionalized membranes demonstrated an elongated spindle-shaped morphology. Conclusion: The functionalization of collagen membranes with Dox promoted an increase in the proliferation and differentiation of osteoblasts. 2023-05-16T08:12:33Z 2023-05-16T08:12:33Z 2023-04 journal article Toledano-Osorio M, de Luna-Bertos E, Toledano M, Manzano-Moreno FJ, García-Recio E, Ruiz C, Osorio R, Sanz M. Doxycycline-doped collagen membranes accelerate in vitro osteoblast proliferation and differentiation. J Periodontal Res. 2023 Apr;58(2):296-307. doi: 10.1111/jre.13091. https://hdl.handle.net/10481/81567 https://doi.org/10.1111/jre.13091 eng http://creativecommons.org/licenses/by-nc-nd/4.0/ embargoed access Attribution-NonCommercial-NoDerivatives 4.0 Internacional Wiley Online Library