Proliferation study and microscopy evaluation on the effects of tannic acid in human fetal osteoblast cell line (hFOB 1.19)

Abstract

Tannic acid (TA) is a phenolic compound that might act directly on osteoblast metabolism. The study was performed to investigate the effects of TA on the proliferation, mineralization, and morphology of human fetal osteoblast cells (hFOB 1.19). The cells were divided into TA‐treated, untreated, and pamidronate‐treated (control drug) groups. Half maximal effective concentration (EC50) values for TA and pamidronate were measured using MTT assay. The EC50 of hFOB 1.19 cells treated with TA was 2.94 M. This concentration was more effective compared to the pamidronate (15.27 M). Cell proliferation assay was performed to compare cell viability from Day 1 until Day 14. The morphology of hFOB 1.19 was observed via inverted microscope and scanning electron microscope. Calcium (Ca) and phosphate (P) were assessed using energy‐dispersive X‐ray (EDX) analysis. Furthermore, the mineralization of hFOB 1.19 was determined by von Kossa staining (P depositions) and Alizarin Red S staining (Ca depositions). The number of cells treated with TA was significantly higher than the two control groups at Day 10 and Day 14. The morphology of cells treated with TA was uniformly fusiform‐shaped with filopodia extensions. Besides, globular‐like structures of deposited minerals were observed in the TA‐treated group. In line with other findings, EDX spectrum analysis confirmed the presence of Ca and P. The cells treated with TA had significantly higher percentage of both minerals at Day 3 and Day 10 compared to the two control groups. In conclusion, TA enhances cell proliferation and causes cell morphology changes, as well as improved mineralization.

Potential Effects of Polyphenols on Osteoblast and Osteoclast Culture

“…It also possesses antioxidant, antimutagenic, and anticarcinogenic activities 67 . TA treatment is more effective in increasing hFOB 1.19 cell proliferation (EC 50 = 2.94 M) than pamidronate (PAM) (EC 50 = 15.27 M), a nitrogen-containing bisphosphonate that is used to inhibit bone resorption 20 . TA treatment also increased the calcium phosphate (Ca/P) molar ratio in a time-dependent manner (day 3 and day 10), which is crucial for the mineralization of the extracellular matrix 20 .…”

“…Studies have been conducted on primary cells, such as primary mouse mesenchymal progenitor cell-derived OBs 17, 18 , primary human osteoblast (Hob) cells, and PMBC-derived OCs 19 . Cell lines include the human fetal osteoblast cell line (hFOB 1.19) 20 , RAW264.7-derived OCs 21 , and MC3T3-E1 OBs 22 . Because OBs and OCs are important during osteogenesis and remodeling, these cells are used for in vitro studies of bone diseases 23, 24 .…”

Synthesis and in Vitro Toxicity Assessment of Different Nano-Calcium Phosphate Nanoparticles

“…The cells were incubated in a 5% CO2 37°C humidified incubator and monitored closely for 24 hours. In this study, Triton-X-100 (1%) was used as a positive control (Control+), while the untreated cells were designed as negative control (Control−) [19][20].…”

Toughening of Bioceramic Composites for Bone Regeneration

“…This helps it in adapting to ever changing biomechanical forces by replacing the old or micro-damaged bone with a new and mechanically stronger bone, thereby preserving bone strength [91]. The bone has a unique ability to shift the intricate balance between osteoclastic and osteoblastic activities depending upon the external stimuli [92][93][94]. Such mechanotransduction signals can amplify the osteoblastic activity, resulting in an enhanced deposition of bone matrix [44].…”