Runt-Related Transcription Factor 2 (Runx2) Measurement in Phytoestrogen-Induced Bone: A Comparison of Western Blot and Immunohistochemistry Methods

Abstract

Estrogen deficiency can contribute to osteoporosis in postmenopausal women. Phytoestrogens are becoming more widely recognized as potential estrogen replacement therapy. The administration of phytoestrogens can cause bone formation, which is marked by an increase in Runx2 expression in osteoblast cells and can be seen using western blot and immunohistochemistry approaches. This review aimed to compare the detection methods of Runx2 in phytoestrogen-induced bone tissue using western blots and immunohistochemistry. Selectivity, sensitivity, processing time, and cost-effectiveness were the parameters that were compared. This review was done by identifying articles in several databases (Google Scholar, PubMed, and Science Direct). The process of selecting the articles used the PRISMA guidelines to create a flowchart with inclusion and exclusion study criteria. Meta-synthesis was done to analyze, identify, and interpret all of the data in the articles systematically. 70 articles in total were obtained from the selection process, with 21 articles being relevant to the topic. The result shows that the selectivity and sensitivity of western blot for detecting Runx2 on tissue were 93.5–100%, respectively, whereas immunohistochemistry selectivity and sensitivity were 45–99.5%, respectively. Compared to immunohistochemistry, western blot can save up to 57.26%. Immunohistochemistry takes 46 hours to process, while Western blot takes 25 hours and 20 minutes. In comparison to immunohistochemistry, the western blot is more selective, sensitive, rapid and affordable for detecting Runx2 in bone tissue.

A Novel Approach Using Reduced Graphene Oxide for the Detection of ALP and RUNX2 Osteogenic Biomarkers

“…Traditionally, the quantification of ALP and RUNX2 has relied on laborious and often expensive techniques, including enzyme-linked immunosorbent assays (ELISAs) [20,21], Western blotting [22,23], and polymerase chain reaction (PCR) [24,25]. These methods, while valuable for research, are impractical for rapid, point-of-care diagnostics or realtime monitoring of osteogenic processes.…”

“…Mahato et al [34] developed an impedimetric biosensor using gold-nano-dendroids and graphene oxide (GO) detecting ALP in human serum samples, Liu et al [35] describe a fluorescent assay based on GO and λ exonuclease for ALP activity, while to the best of our knowledge, RUNX2 detection is still performed using classical techniques like quantitative PCR (qPCR) [36], Western blot [37], and immunohistochemistry [38], while no progress has been made regarding the development of a novel electrochemical biosensor to detect this biomarker. Ma'arif et al [22] compared two techniques used for RUNX2 detection in bone tissue, i.e., Western blot and immunohistochemistry, investigating 70 articles, and concluded that the Western blot method is superior in terms of sensitivity, selectivity, cost, and detection time. However, neither of these techniques gives a fast response, the Western blot method having a process time over 25 h, while immunohistochemistry takes over 46 h to generate a result.…”