Prolotherapy upregulates Fibroblast Growth Factor‐2 (FGF‐2) and Insulin‐like Growth Factor‐1 (IGF‐1) mRNA which may increase chondrocyte proliferation and mediates osteoarthritic inflammation

Abstract

Osteoarthritis is a highly prevalent disease that is progressively generated and has no known cure. Current treatments, such as opioids, have highly adverse effects. A more effective treatment can be the form of proliferative therapy (prolotherapy), a complementary medical intervention with promising clinical evidence. The purpose of the present study is to investigate the molecular basis of prolotherapy as a treatment option for osteoarthritis. Specifically, the study investigates whether prolotherapy upregulates proliferation‐enhancing cytokines (FGF‐2 and IGF‐1) in chondroprogenitor cells. In vitro stimulation of chondroprogenitor cells was carried out with prolotherapy treatment followed by qRT‐PCR quantification to determine the mRNA expression of the genes FGF‐2, IGF‐1, CCND‐1, TGF‐β1, AKT, STAT1, and BMP2. Results indicated a likely upregulation of cytokines that enhanced cell proliferation in chondroprogenitor cells with statistically higher levels of expression for FGF‐2 (p<0.001, p<0.05, p<0.01), IGF‐1 (p<0.001), and CCND‐1 (p<0.05) relative to controls. Results in conjunction with prolotherapy and molecular‐focused inflammation theory suggest that prolotherapy has the potential to transform chondroprogenitor cells that are dysregulated by osteoarthritic inflammation. This would contribute to cartilage degradation back into chondroprogenitor cells that proliferate and balance catabolic/anabolic roles to repair and maintain cartilage.P2G (1.5%) upregulates FGF‐2 mRNA expression in chondroprogenitor cells. A Exploratory Trial 1 suggests P2G effects FGF‐2 gene expression (Welch’s one‐way ANOVA, p<0.01). B Confirmatory Trial 2 indicates that relative to time‐specific controls, chondrocytes treated with P2G exhibit increased levels of FGF‐2 at hours 24, 30, and 38 (Welch’s t‐tests yield p<0.001, p<0.05, and p<0.01). qRT‐PCR quantifies mRNA expression. NS p>0.05, * p<0.05, ** p<0.01, *** p<0.001Figure 1P2G (1.5%) upregulates CCND‐1 (Cyclin D) mRNA expression at hour 30 in chondroprogenitor cells in the confirmatory Trial 2 (directional Welch’s t‐test yields p<0.05). CCND‐1 mRNA upregulation at hour 30 suggests subsequent entry chondroprogenitor cell proliferation. qRT‐PCR quantifies mRNA expression. NS p>0.05, * p<0.05, ** p<0.01, *** p<0.001Figure 2