Pressurized carbon dioxide combined with aqueous ethanol as cosolvent induces efficient delipidation of porcine retina for their use as bioscaffolds

Abstract

Retina damage results in both health and social problems. To treat retina damage, extracellular matrix technology has gained interest in the scientist community. Bioscaffolds production is based in the use of a `cleaned` tissue, free of undesired cellular components, for further recellularization using healthy cells. Removal of lipids is an important step to improve both deand recellularization. This work shows, for the first time, the efficiency of pressurized carbon dioxide (CO2) fluids for the delipidation of porcine retina tissue. Pressurized CO2-EtOH-H2O (0.87 XCO2) at 300 bar and 37 °C for 1h completely removed the 28 lipid species found in untreated retina (classified as phosphatidylcholines, phosphatidylethanolamines and sphingomyelins). When the ethanolic mixture was replaced by limonene at same conditions, lipid content decreased by more than a 50%, indicating reasonably good solubility of retina lipids in pressurized CO2limonene; however, no delipidation was observed using neat supercritical CO2. In conclusion, pressurize CO2 technologies can be applied for delipidation of retina. Whether the resulting bioscaffold can be recellularized and used in transplantation remain to be shown.

Pressurized carbon dioxide as a potential tool for decellularization of pulmonary arteries for transplant purposes

“…3). The extensive delipidation observed for pressurized CO 2 -EtOH-H 2 O, compared to scCO 2 and pressurized CO 2 -limonene, are in line with results obtained for porcine retina 32 . The relative permittivity of CO 2 is low, which makes it appropriate to dissolve mostly non-polar compounds of low molecular weight 41 .…”

“…5d), in line with the unexpected results from the gravimetric analysis. Similar trends have been observed for other tissue treated with pressurized CO 2 -based fluids 32 . The reason for this observation may be a result of the pressurized CO 2 (at both supercritical or pressurized state) impacting on FA availability and facilitating subsequent extraction with dichloromethane.…”

“…S4), however not in the supercritical regime. Such fluids are rarely studied 31 , but they offer an even higher range of polarizability than supercritical mixtures, expanding the type of extractable lipids 32 . Mass transfer properties of such pressurized fluids are also modified with respect to the neat cosolvent, in favor of better extraction power.…”

“…2). Samples were then treated using an in-house constructed equipment, as previously reported 32 . Briefly, the sample holder was placed in a stainless-steel extraction vessel with a volume of 80 mL (Ångström Laboratory, Uppsala www.nature.com/scientificreports www.nature.com/scientificreports/ University, Uppsala, Sweden), mounted on top of a magnetic stirrer (VWR, Leuven, Belgium) inside a GC-oven (HP 5890 GC, Hewlett-Packard Co., Palo Alto, CA).…”

Decellularized extracellular matrix scaffolds: Recent trends and emerging strategies in tissue engineering

“… [ 14 , [49] , [50] , [51] , [52] , 57 , [76] , [77] , [78] , [79] , [80] , [81] ] Sophisticated to operate the perfusion devices Scraping Mechanically remove cells from ECM surface Damage underlying basement membranes. [ 82 ] Need standards for controlling mechanical force Sonication Generate acoustic cavitation bubbles, induce shear stress effect, and thus rupture the cell membrane; Ease and assist the penetration of agents by emitting the vibration, and help cellular debris removal High power or longer duration of sonication could disrupt the main structural fibers and produce adverse effects on the vascular tissues [ [83] , [84] , [85] ] Supercritical fluids Facilitate chemical exposure, and lead to cell removal Not yet widely used [ [86] , [87] , [88] ] Pressure gradient/Convective flow Burst cells, and aid in the delivery of solutions to force cellular components and soluble proteins out of tissues Make solutions difficult to penetrate ECM due to the constantly high pressure [ [89] , [90] , [91] , [92] , [93] ] Pressurization Result in cell lysis Denature the proteins, and disrupt the ECM structure by formed ice crystals. …”