Comparison of the Anabolic Effects of Reported Osteogenic Compounds on Human Mesenchymal Progenitor-Derived Osteoblasts

Abstract

There is variability in the reported effects of compounds on osteoblasts arising from differences in experimental design and choice of cell type/origin. This makes it difficult to discern a compound’s action outside its original study and compare efficacy between compounds. Here, we investigated five compounds frequently reported as anabolic for osteoblasts (17β-estradiol (oestrogen), icariin, lactoferrin, lithium chloride, and menaquinone-4 (MK-4)) on human mesenchymal progenitors to assess their potential for bone tissue engineering with the aim of identifying a potential alternative to expensive recombinant growth factors such as bone morphogenetic protein 2 (BMP-2). Experiments were performed using the same culture conditions to allow direct comparison. The concentrations of compounds spanned two orders of magnitude to encompass the reported efficacious range and were applied continuously for 22 days. The effects on the proliferation (resazurin reduction and DNA quantification), osteogenic differentiation (alkaline phosphatase (ALP) activity), and mineralised matrix deposition (calcium and collagen quantification) were assessed. Of these compounds, only 10 µM MK-4 stimulated a significant anabolic response with 50% greater calcium deposition. Oestrogen and icariin had no significant effects, with the exception of 1 µM icariin, which increased the metabolic activity on days 8 and 22. 1000 µg/mL of lactoferrin and 10 mM lithium chloride both significantly reduced the mineralised matrix deposition in comparison to the vehicle control, despite the ALP activity being higher in lithium chloride-treated cells at day 15. This demonstrates that MK-4 is the most powerful stimulant of bone formation in hES-MPs of the compounds investigated, highlighting its potential in bone tissue engineering as a method of promoting bone formation, as well as its prospective use as an osteoporosis treatment.

Effects of PIN on Osteoblast Differentiation and Matrix Mineralization through Runt-Related Transcription Factor

“…Osteoblasts, specialized cells derived from mesenchymal stem cells (MSCs), are essential for bone formation and maintenance, and osteoblast dysfunction causes pathological fragility and diseases such as osteoporosis and periodontitis [ 3 ]. Rovert et al reported the anabolic effects of osteogenic compounds on human mesenchymal progenitor-derived osteoblasts and suggested the possibility of bone tissue engineering and bone diseases [ 4 ]. Anabolic treatments for osteoporosis increase the differentiation and maturation of osteoblast by altering bone metabolism [ 5 , 6 ].…”

Combined Porogen Leaching and Emulsion Templating to produce Bone Tissue Engineering Scaffolds

“…Calcium and collagen deposition were quantified on days 7 and 14 by alizarin red S (ARS) and direct red 80 (DR80), respectively, as previously reported Owen et al . [ 48 ] Briefly, the samples were rinsed twice in PBS then fixed by immersion in 3.7% formaldehyde for 30 min. Scaffolds were rinsed twice in diH 2 O, and then submerged in 1 w/v% ARS in diH 2 O for 30 min to stain for calcium.…”

Recent Advances in Regenerative Tissue Fabrication: Tools, Materials, and Microenvironment in Hierarchical Aspects

“…The incorporation of a 2D cell culture into a microfluidic device has led to 3D approaches with sophisticated designs that mimic the mechanical and dynamic environments of native tissue. [ 226 ] Fabrication of complex, anisotropic, hierarchical macrostructures has been achieved by combining and integrating different technologies and materials. Bahmaee et al used a fabrication method based on a polymerized high‐internal‐phase emulsion (polyHIPE) to advance an osteogenesis‐on‐a‐chip device that incorporated a 3D environment and fluid shear stresses.…”