Biomimetic Collagen Membranes as Drug Carriers of Geranylgeraniol to Counteract the Effect of Zoledronate

Francisco Javier Manzano-Moreno; Elvira de Luna-Bertos; Manuel Toledano-Osorio; Paula Urbano-Arroyo; Concepción Ruiz; Manuel Toledano; Raquel Osorio

doi:10.3390/biomimetics9010004

Biomimetic Collagen Membranes as Drug Carriers of Geranylgeraniol to Counteract the Effect of Zoledronate

Biomimetics (Basel). 2023 Dec 22;9(1):4. doi: 10.3390/biomimetics9010004.

Authors

Francisco Javier Manzano-Moreno^{1

2}, Elvira de Luna-Bertos^{2

3}, Manuel Toledano-Osorio⁴, Paula Urbano-Arroyo⁴, Concepción Ruiz^{2

3

5}, Manuel Toledano^{2

4}, Raquel Osorio^{2

4}

Affiliations

¹ Biomedical Group (BIO277), Department of Stomatology, School of Dentistry, University of Granada, 18071 Granada, Spain.
² Instituto Investigación Biosanitaria, ibs. Granada, 18012 Granada, Spain.
³ Biomedical Group (BIO277), Department of Nursing, Faculty of Health Sciences, University of Granada, 18016 Granada, Spain.
⁴ Faculty of Dentistry, University of Granada, Colegio Máximo de Cartuja s/n, 18071 Granada, Spain.
⁵ Institute of Neuroscience, University of Granada, Centro de Investigación Biomédica (CIBM), Parque de Tecnológico de la Salud (PTS), 18071 Granada, Spain.

Abstract

To counteract the effect of zoledronate and decrease the risk of osteonecrosis of the jaw (BRONJ) development in patients undergoing guided bone regeneration surgery, the use of geranylgeraniol (GGOH) has been proposed. Collagen membranes may act as biomimetical drug carriers. The objective of this study was to determine the capacity of collagen-based membranes doped with GGOH to revert the negative impact of zoledronate on the growth and differentiation of human osteoblasts. MG-63 cells were cultured on collagen membranes. Two groups were established: (1) undoped membranes and (2) membranes doped with geranylgeraniol. Osteoblasts were cultured with or without zoledronate (50 μM). Cell proliferation was evaluated at 48 h using the MTT colorimetric method. Differentiation was tested by staining mineralization nodules with alizarin red and by gene expression analysis of bone morphogenetic proteins 2 and 7, alkaline phosphatase (ALP), bone morphogenetic proteins 2 and 7 (BMP-2 and BMP-7), type I collagen (Col-I), osterix (OSX), osteocalcin (OSC), osteoprotegerin (OPG), receptor for RANK (RANKL), runt-related transcription factor 2 (Runx-2), TGF-β1 and TGF-β receptors (TGF-βR1, TGF-βR2, and TGF-βR3), and vascular endothelial growth factor (VEGF) with real-time PCR. One-way ANOVA or Kruskal-Wallis and post hoc Bonferroni tests were applied (p < 0.05). Scanning electron microscopy (SEM) observations were also performed. Treatment of osteoblasts with 50 μM zoledronate produced a significant decrease in cell proliferation, mineralization capacity, and gene expression of several differentiation markers if compared to the control (p < 0.001). When osteoblasts were treated with zoledronate and cultured on GGOH-doped membranes, these variables were, in general, similar to the control group (p > 0.05). GGOH applied on collagen membranes is able to reverse the negative impact of zoledronate on the proliferation, differentiation, and gene expression of different osteoblasts' markers.

Keywords: collagen membranes; geranylgeraniol; osteoblast; zoledronate.

Grants and funding

Grant PID2020-114694RB-I00 funded by MCIN/AEI 10.13039/501100011033. M.T-O. was fellow FPU of Ministry of Universities [grant FPU20/00450]. Authors want to thank Klockner S.A. for providing the collagen membranes.