Open AccessProlotherapy agent P2G is associated with upregulation of fibroblast growth factor‐2 genetic expression in vitro
Author(s) -
Johnston Elisha,
Emani Chandrakanth,
Kochan Andrew,
Ghebrehawariat Kidane,
Tyburski John,
Johnston Michael,
Rabago David
Publication year - 2020
Publication title -
journal of experimental orthopaedics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.597
H-Index - 18
ISSN - 2197-1153
DOI - 10.1186/s40634-020-00312-z
Subject(s) - downregulation and upregulation , chondrocyte , fibroblast growth factor , cartilage , medicine , prolotherapy , immunology , endocrinology , chemistry , pathology , anatomy , biochemistry , gene , receptor , alternative medicine
Purpose Osteoarthritis (OA) is a prevalent, progressively degenerative disease. Researchers have rigorously documented clinical improvement in participants receiving prolotherapy for OA. The mechanism of action is unknown; therefore, basic science studies are required. One hypothesized mechanism is that prolotherapy stimulates tissue proliferation, including that of cartilage. Accordingly, this in vitro study examines whether the prolotherapy agent phenol‐glycerin‐glucose (P2G) is associated with upregulation of proliferation‐enhancing cytokines, primarily fibroblast growth factor‐2 (FGF‐2). Methods Murine MC3T3‐E1 cells were cultured in a nonconfluent state to retain an undifferentiated osteochondroprogenic status. A limitation of MC3T3‐E1 cells is that they do not fully reproduce primary human chondrocyte phenotypes; however, they are useful for modeling cartilage regeneration in vitro due to their greater phenotypic stability than primary cells. Two experiments were conducted: one in duplicate and one in triplicate. Treatment consisted of phenol‐glycerin‐glucose (P2G, final concentration of 1.5%). The results were assessed by quantitative Reverse Transcriptase‐Polymerase Chain Reaction (qRT‐PCR) to detect mRNA expression of the FGF‐2, IGF‐1, CCND‐1 (Cyclin‐D), TGF‐β1, AKT, STAT1, and BMP2 genes. Results P2G ‐ treated preosteoblasts expressed higher levels of FGF‐2 than water controls (hour 24, p < 0.001; hour 30, p < 0.05; hour 38, p < 0.01). Additionally, CCND‐1 upregulation was observed ( p < 0.05), possibly as a cellular response to FGF‐2 upregulation. Conclusions The prolotherapy agent P2G appears to be associated with upregulation of the cartilage cell proliferation enhancer cytokine FGF‐2, suggesting an independent effect of P2G consistent with clinical evidence. Further study investigating the effect of prolotherapy agents on cellular proliferation and cartilage regeneration is warranted.