Released fibroblast growth factor18 from a collagen membrane induces osteoblastic activity involved with downregulation of miR-133a and miR-135a

Abstract

We have developed a unique delivery system of growth factors using collagen membranes (CMs) to induce bone regeneration. We hypothesized that fibroblast growth factor18 (FGF-18), a pleiotropic protein that stimulates proliferation in several tissues, can be a good candidate to use our delivery system for bone regeneration. Cell viability, cell proliferation, alkaline phosphatase activity, mineralization, and marker gene expression of osteoblastic differentiation were evaluated after mouse preosteoblasts were cultured with a CM containing FGF-18, a CM containing platelet-derived growth factor, or a CM alone. Furthermore, expression of microRNA, especially miR-133a and miR-135a involving inhibition of osteogenic factors, was measured in preosteoblasts with CM/FGF-18 or CM alone. A sustained release of FGF-18 from the CM was observed over 21 days. CM/FGF-18 significantly promoted cell proliferation, alkaline phosphatase activity, and mineralization compared to CM alone. Gene expression of type I collagen, runt-related transcription factor 2, osteocalcin, Smad5, and osteopontin was significantly upregulated in CM/FGF-18 compared to CM alone, and similar to CM/platelet-derived growth factor. Additionally, CM/FGF-18 downregulated expression of miR-133a and miR-135a. These results suggested that released FGF-18 from a CM promotes osteoblastic activity involved with downregulation of miR-133a and miR-135a.

RETRACTED ARTICLE: Bone Marrow Mesenchymal Stem Cell-Derived Exosomal MicroRNA-133a Restrains Myocardial Fibrosis and Epithelial–Mesenchymal Transition in Viral Myocarditis Rats Through Suppressing MAML1

“…Another finding in our study was that up-regulated exosomal miR-133a decreased CVF, reduced the expression of collagen I and collagen III in rats with VMC. A article has elucidated that released fibroblast growth factor-18 from a collagen membrane causes osteoblastic activity participating in down-regulated miR-133a [ 31 ]. In vitro excessive expression of miR-133a depresses cardiomyocyte hypertrophy and reduces collagen expression [ 32 ], as evidenced in another study.…”

Growth Factor Delivery Using a Collagen Membrane for Bone Tissue Regeneration

“…In the case of drug delivery for bone regeneration, this type of kinetics is desirable because it promotes a prolonged duration for the development of new bone. In the CMs with GFs unit developed in our study, this release kinetics has been confirmed for up to 14~21 days [96][97][98][99][100][101]. The total cumulative amount of sustained release ranged from 30% to ≈100%.…”

“…4.5. CMs with FGF-18 (CM/FGF-18) [99] We have created a novel delivery system based on proprietary growth factor with CMs to induce bone regeneration. We hypothesized that FGF-18, a multifaceted protein that elevates proliferation in multiple tissues, could be a good candidate for bone reconstruction using our delivery system.…”

New developments in the biology of fibroblast growth factors

“…miR‐133a and miR‐135a are involved in the inhibition of osteogenic factors including runt‐related transcription factor 2 (Runx2) and Smad5. Recombinant FGF18, released from collagen membranes, promoted osteoblastic activity by decreasing the expression of miR‐133a and miR‐135a to allow increased expression of osteogenic factors (Imamura et al, 2018). Cyclic mechanical loading‐induced osteogenic differentiation of human periodontal ligament cells and decreased expression of endogenous miR‐195‐5p.…”