Cell Culture in a Hyperbaric Chamber: A Research Model to Study the Effects of Hyperbarism (Hyperbaric Pressure) on Bone Cell Culture.

The hyperbaric environment, to which many categories of workers are exposed, can provoke injuries that can lead to various types of disorders. A major part of the studies aiming to explore the causes/effects leading to these injuries are conducted in vivo. In the present manuscript, we describe the effects on osteoblast cell cultures stressed in a hyperbaric purpose-built chamber, using an in vitro model to analyze the affected pathways. A hyperbaric chamber for cell cultures was constructed by adapting a pressurized test chamber originally designed for technical use. The MG-63 cell line and human primary osteoblasts were placed into this chamber at different atm and exposure times, at 37 °C. After treatment, the chamber was depressurized by performing controlled decompression stops. Then, the pro-inflammatory cytokines and bone tissue biomarker expression were analyzed. The stress conditions induced the overexpression of pro-inflammatory cytokines, such as IL-6, IL-1β, and TNF-α, along with reactive oxygen species release. Moreover, the alteration of bone tissue marker production was observed. In particular, the increase in Receptor Activator of NF-κB Ligand (RANKL) and the decrease in Osteoprotegerin (OPG) were detected. Further modulation was observed regarding other biomarkers, Alkaline phosphatase, Osteocalcin, Bone Morphogenetic Protein-2, and mainly Collagen type I, all of which were downregulated by treatment. Taken together, these findings account for certain illnesses, such as dysbaric osteonecrosis, diagnosed in workers exposed to a hyperbaric environment. Inflammation induced by this kind of stress affects several factors involved in bone tissue homeostasis, leading to bone injuries, which are among the typical disorders observed in divers.