Synthesized Nanorods Hydroxyapatite by Microwave-Assisted Technology for In Vitro Osteoporotic Bone Regeneration through Wnt/β-Catenin Pathway

Abstract

This research presents an optimal and inexpensive, without any additives, method for the synthesis and sintering of hydroxyapatite (HA) by microwave-assisted technology (MAT) furnace. The target sintering temperature of the furnace (1100 ℃) was held for one and two hours for conventional sintering. With regard to the microwave hybrid sintering, it was held at 100%MW for 20 and 30 min. FTIR, XRD, TGA, SEM/EDS, and TEM were assessed to determine HA phase composition, and structural as well as thermal decomposition behavior. The in vitro effects of sintered HA discs on cultured aged mice-isolated osteoblast cells and hydrocortisone-induced osteoclast cells were assessed by measuring ALP, osteocalcin, TRAP, calcium, and Alizarin red S staining. Moreover, their effects on cell differentiation (CD90 and CD 105 and PARR- ɣ) and death markers (GSK3b, MAPK, and β-catenin) were evaluated. The results demonstrate the production of ≈35 nm crystal-sized pure hydroxyapatite nanorod-like particles with a high degree of crystallinity and no impurities as required for biomedical application. HA increased osteogenesis (ALP, osteocalcin, and calcium) markers and decreased cell resorption markers. In addition, HA nanorods reversed the effect of cortisone on cell differentiation and death markers. In conclusion, microwave hybrid sintered HA is a potential nanomaterial for osteoporotic bone regeneration as HA reversed the cortisone adverse effect on osteoblast cell death through canonical and non-canonical pathways.

Cellulose Acetate Nanofibers: Incorporating Hydroxyapatite (HA), HA/Berberine or HA/Moghat Composites, as Scaffolds to Enhance In Vitro Osteoporotic Bone Regeneration

“…Activation of this pathway induces osteoblasts cell death. Despite the direct HCT effect on Wnt/β-catenin pathway, inflammation microenvironment induced β-catenin accumulation which increased also when adipocytes number was increased in order to suppresses the non-canonical Wnt/Ca( 2+ ) pathway thus leading to cell proliferation positive increment that associated with osteogenic differentiation reduction [ 20 ]. Treatment with HA/BER or HA/ME carried on CA13 fiber improved the tested parameters but did not normalize them.…”

“…Cells were isolated from 0 to 5 days old neonatal male Wister albino rats according to Shaban, N.Z. et al 2021 our previous study [ 20 ]. The parietal and occipital bones were dissected and washed with phosphate buffered saline (PBS).…”

“…Then, 0.5 mg/mL hydrocortisone (HCT) was added and every 3–4 days the media was changed. After 21 days of culture, osteoclast-like cells formation were observed [ 20 ].…”

“…The nanorod-like HA is better than other forms because they not only reduce bone loss but also increase cortical bone thickness, and they mimic the natural bone HA nanocrystals (a plate-like shape with a length of 30–200 nm and a thickness of 2–7 nm) [ 18 ]. Nanorod hydroxyapatite enhanced osteoblast functions proliferation, alkaline phosphates, calcium synthesis, bone formation transcription factors production that correlate with decreased β-catenin accumulation leading to activation of the Wnt/β-catenin pathway [ 20 ]. The resorptive activity of osteoclasts cultured with nanorods HA was weaker than that of control osteoclastic cells.…”

“…In addition, cell adhesion and proliferation were enhanced by the presence of HA. Therefore, nanorods hydroxyapatite may serve as a valuable scaffold for osteoporotic bone regeneration [ 20 ].…”