Interleukin 6 and Interleukin 17a Enhance Proliferation and Differentiation of Murine Osteoblast and Human Foetal Osteoblast Cell Lines

Introduction: Cytokines have been gaining great focus due to their role in enhancing osseointegration as well as their potential in bone reconstruction. Osseointegration often faces complications in its compatibility with the implant due to rejection by the recipients own immune system. Therefore, extensive studies are being carried out to enhance osteoblast development to minimize such complication. The aim of this study was to determine the effect of different concentrations of Interleukin 6 (IL-6) and Interleukin 17a (IL-17A) in the proliferation and differentiation of murine and human osteoblasts. Methods: Various concentrations (5, 10, 25 and 50 ng/ml) of rIL-6 and rIL-17A were tested on both murine osteoblast (MC3T3-E1) and human feotal osteoblast (hFOB) cell lines using [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] (MTS) and alkaline phosphatise (ALP) assays. MTS was carried out at 24, 48, 72, 96 and 120 hours while ALP assay was done on day 1, 3, 7, 10 and 14. Results: MC3T3-E1 cells showed steadier proliferation and differentiation compared to hFOB. Both cell lines expressed responses in dose-dependent manner. The concentration of 10ng for IL-6 and IL-17A in the case of MC3T3-E1 cell line was found to be the most suitable for further studies. Conclusion: IL-6 and IL-17A enhance proliferation and ALP activity of both MC3T3-E1 and hFOB cell lines.

A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway

The skeletal system plays an important role in the development and maturation process. Through the bone remodeling process, 10% of the skeletal system is renewed every year. Osteoblasts and osteoclasts are two major bone cells that are involved in the development of the skeletal system, and their activity is kept in balance. An imbalance between their activities can lead to diseases such as osteoporosis that are characterized by significant bone loss due to the overactivity of bone-resorbing osteoclasts. Our laboratory has developed a novel peptide, CK2.3, which works as both an anabolic and anti-resorptive agent to induce bone formation and prevent bone loss. We previously reported that CK2.3 mediated mineralization and osteoblast development through the SMAD, ERK, and AKT signaling pathways. In this study, we demonstrated the mechanism by which CK2.3 inhibits osteoclast development. We showed that the inhibition of MEK by the U0126 inhibitor rescued the osteoclast development of RAW264.7 induced by RANKL in a co-culture system with CK2.3. We observed that CK2.3 induced ERK activation and BMPRIa expression on Day 1 after stimulation with CK2.3. While CK2.3 was previously reported to induce the SMAD signaling pathway in osteoblast development, we did not observe any changes in SMAD activation in osteoclast development with CK2.3 stimulation. Understanding the mechanism by which CK2.3 inhibits osteoclast development will allow CK2.3 to be developed as a new treatment for osteoporosis.