Angiogenesis and hyperbaric oxygen in the chick embryo chorioallantoic membrane

Abstract

Hyperbaric Oxygen Therapy (HBOT) is increasingly applied in different areas of medical practice. The oxy-hyperbarism effects are not well understood in cancer malignancy. One unique feature of cancer is the presence of hypoxic regions that are insensitive to conventional therapies. It is possible to alter the hypoxic state and produce reactive oxygen species for better treatment outcome by HBOT. In the present study, we determined the effects of HBOT on angiogenesis, a signature of cancer progression, by using the chick chorioallantoic membrane (CAM) in vivo assy. CAMs were exposed to 2.0 ATA (atmospheres absolute) for 30 min of hyperbaric oxygen on the 6 th and 7 th days of incubation (ED6, ED7). On the 10-11 th day of incubation, CAMs were excised from eggs, fixed and analysed using APERIO ImageScope software. HBOT outcomes were evaluated quantifying the volumetric area occupied by blood vessels and calculating the number of blood vessel ramifications. Results indicated that CAMs treated at ED6 and ED7 had a significantly higher CAM vascularization and an increased number of blood vessel ramifications (+82% higher for ED6) compared to untreated CAMs (ED6=63.3±2.5 and ED7=57.7±5.5 vs. CTRL=34.7±2.5). Thus, HBOT induces an angiogenic response in treated CAMs through a classic sprouting mechanism.

Hyperbaric Oxygen Increases Stem Cell Proliferation, Angiogenesis and Wound-Healing Ability of WJ-MSCs in Diabetic Mice

“…Indeed, Chan and Burggren (2005) have shown that CAM mass is unchanged by hypoxia experienced during early or mid-development, but CAM mass at E18 is greatly increased by hypoxia. Despite these contrasting results, the present study is in agreement with that presented by Montecorboli et al (2015) where significantly higher CAM vascularization and an increased number of blood vessels were found for E6 and E7 subjected to HBOT (protocol: 2.0 ATA, for 30 min).…”

“…In the literature, several studies extensively explore the effects of hypoxia and hyperoxia on CAM development. However, most of these studies (with the exception of Montecorboli et al, 2015 ) do not evaluate both the effects of pressure and oxygen concentration on embryonic development ( Dusseau and Hutchins, 1988 ; Burton and Palmer, 1992 ; Druyan and Levi, 2012 ). Interestingly, there is conflicting evidence of the effect of ambient oxygen on vascular development.…”

Scleral ossicles: angiogenic scaffolds, a novel biomaterial for regenerative medicine applications

“…64,65 Because of its easy accessibility, the chicken CAM (frequently used as an experimental model, also as an alternative to the animal model, as just mentioned) has been used, in the present study, for the investigation of the angiogenic potential of SO. [66][67][68] Moreover, in the development stages used in this experimentation (from day 8 to day 12) the immunocompetent chicken system is not fully developed and for this reason it is not responsive to an exogenous material. 69 The angiogenic potential of the SO was evaluated by the CAM assay, incubating firstly SO and subsequently media conditioned by SOs.…”

Cell Culture Effects of Altered Oxygen Levels and Hyperbaric Treatment In Vitro

“…The mechanisms of actions needed further investigations, briefly NOS levels were increased in genomic and protein levels as well [20]. In-depth micro-array analyses of microvascular endothelial cells' genome proved huge impact of HBOT on angiogenesis-related gene expressions [21]. In these studies, HBOT dramatically increased tube formation capacity of endothelial cells on Matrigel [22].…”