Methods: Serial dilutions of perfluorooctanoid acid, 1H,1H,7H-dodecafluoro-1- heptanol, 1H,1H,1H,2H,2H-Perfluorooctane, 1H perfluorooctan; ethylbenzene, paraxylene, perfluotobutyfurane, and hexafluoro-1,2,3,4-tetrachlorobutane were tested by direct contact cytotoxicity test using BALB3T3 and ARPE19 cell lines, after application on 59% of the area for 24 h.

Results: Traces of paraxylene, ethylbenzene, and PFOA (≤30ppm) induced severe toxicity in both cell lines. Hexafluoro-1,2,3,4-tetrachlorobutane and H,1H,7H-Dodecafluoro-1-heptanol were cytotoxic at approximately 10000 ppm. 1H perfluorooctan was cytotoxic at approximately 60000 ppm. High concentrations of 1H,1H,1H,2H,2H-Perfluorooctane and perfluotobutyfurane were not cytotoxic.

Conclusions: Six out of eight perfluorocarbon manufacturing process residuals showed cytotoxicity in the direct contact test according to ISO 10993-5. Paraxylene, ethylbenzene, and PFOA were the most cytotoxic compounds. 1H,1H,1H,2H,2H-Perfluorooctane and perfluotobutyfurane were not cytotoxic.

Methods: BALB3T3 and ARPE19 cell lines were cultured in 96-well plates for direct contact cytotoxicity test of 22%, 59%, and 83% contact areas and 2.5, 12, and 24 h contact times. Cell morphology was graded by light microscopy. Cell viability was quantified by MTT assay in ARPE19 cells and neutral red uptake viability assay for BALB3T3. 1-H perfluorooctane (1H PFO) and purified perfluoro-n-octane (PFO) were used as positive and negative controls, respectively.

Results: Qualitative evaluation showed that positive control induced the presence of severe reactivity zones, resulting in 2.2 to 3.0 grading score, and cytotoxicity according to ISO 10993-5 in all tested conditions. Quantitative evaluation of 1H PFO applied on 22%, 59%, and 83% contact areas corresponded to 14–26%, 84–96%, and 86–99% cell mortality ranges, respectively. No cytotoxicity according to ISO 10993-5 was detected in tested cell lines, when 1H PFO was applied on a 22% area at the tested time intervals. The negative control was not cytotoxic with both approaches in all tested conditions.

Conclusions: Direct contact cytotoxicity test according to ISO 10993-5 of PFCLs was validated quantitatively and qualitatively, using ARPE19 and BALB 3T3 cell lines and covering 59% of the cell surface areas for 24 h of contact time. Test conditions using a smaller contact area could not be validated.

Purpose: The vitreous cortex, epiretinal membrane (ERM), and inner limiting membrane (ILM) are transparent tissues and are then difficult to visualize. Staining these structures can increase the efficiency of a nontraumatic removal.

Methods: The surgeon performs a partial core vitrectomy and induces a posterior vitreous detachment. The vital dye is then injected into the retrohyaloid space in balanced salt solution (BSS). The dyes used are TWIN (AL.CHI.MI.A. Srl, Padova, Italy), MembraneBlue-Dual (DORC International, Zuidland, The Netherlands), and Doubledyne (Alfa Intes, Casoria, Italy). The surgeon can complete the vitrectomy and gradually aspirate the dye with a probe. Once the vitrectomy is complete, the surgeon can perform the peeling of the ERM without the need to reinject the vital dye over the macula.

Results: The presence of the dye over the macula facilitates the visualization of the vitreous cortex by blocking the red reflex and increasing the contrast power of the coaxial light probe during the vitrectomy. This allows a negative staining of the vitreous because the dye acts by increasing the visibility of the surrounding BSS and not the vitreous itself.

Conclusions: We describe a new chromovitrectomy technique using the same dye to increase the visualization of the vitreous, posterior hyaloid, ERM, and ILM.