Enhanced osteogenic effect in reduced BMP-2 doses with siNoggin transfected pre-osteoblasts in 3D silk scaffolds


Fuerkaiti S. N., ÇAKMAK A. S., KARAASLAN İ. Ç., GÜMÜŞDERELİOĞLU M.

INTERNATIONAL JOURNAL OF PHARMACEUTICS, vol.612, 2022 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 612
  • Publication Date: 2022
  • Doi Number: 10.1016/j.ijpharm.2021.121352
  • Journal Name: INTERNATIONAL JOURNAL OF PHARMACEUTICS
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, EMBASE, International Pharmaceutical Abstracts, MEDLINE, Veterinary Science Database
  • Keywords: siRNA, Noggin, BMP-2, Silk scaffold, Bone tissue engineering, BONE MORPHOGENETIC PROTEINS, IN-VIVO, BIOMATERIAL SCAFFOLDS, RNA INTERFERENCE, SIRNA, DIFFERENTIATION, NOGGIN, DELIVERY, HYDROGELS, DEGRADATION
  • Hacettepe University Affiliated: Yes

Abstract

Bone morphogenetic proteins (BMPs), especially BMP-2, are being increasingly used in bone tissue engineering due to its osteo-inductive effects. Although recombinant human BMP-2 (rhBMP-2) was approved by Food and Drug Administration (FDA) to use for bone repair, its high doses cause undesired side effects. In order to reduce the BMP-2 dose for enhanced osteogenic differentiation, in this study we decided to suppress the synthesis of Noggin protein, the primary antagonist of BMP-2, on the MC3T3-E1 cells using Noggin targeted small interfering RNA (siRNA). Unlike other studies, Noggin siRNA (siNoggin) transfected cells were seeded on silk scaffolds, and osteogenic differentiation was investigated for a long-term period (21 days) with MTT, qPCR, SEM/EDS, and histological analysis. Besides, siNoggin transfected MC3T3-E1 cells were evaluated as a new cell source for tissue engineering studies. It was determined that Nog gene expression was suppressed in the siNoggin group and Ocn gene expression increased 5-fold compared to the control group (*p < 0.05). The osteogenic effect of BMP-2 was clearly observed in siNoggin transfected cells. According to the SEM/EDS analysis, the siNoggin group has mineral structures clustered on cells, which contain intense Ca and P elements. Histological staining showed that the siNoggin group has a more intense mineralized area than that of the control group. In conclusion, this study indicated that Noggin silencing by siRNA induces osteogenic differentiation in reduced BMP-2 doses for scaffold based bone regeneration. This non-gene integration strategy has as a safe therapeutic potential to enhance tissue regeneration.