This constant high rate of ROS production leads rapidly to extreme macromolecular oxidation, here it is observed in the AOPP and MDA detected after 3 h in samples treated with bare P25TiO₂NPs (Fig. 6, Fig. 7). Macromolecular oxidation includes, among others, both protein and lipid oxidation. The ROS causes protein oxidation by direct reaction or indirect reactions with secondary by-products of oxidative stress. Protein fragmentation or cross-linkages could be produced after the oxidation of amino acid side chains and protein backbones. These and later dityrosine-containing protein products formed during excessive production of oxidants are known as advanced oxidation protein products (AOPP). They absorb at 340 nm and are used to estimate the damage to structural cell amino acids. Lipid oxidation is detected by the conjugation of oxidized polyunsaturated lipids with thiobarbituric acid, forming a molecule that absorbs light at 532 nm. Polyunsaturated lipids are oxidized as a result of a free-radical-mediated chain of reactions. The most exposed targets are usually membrane lipids. The macromolecular damage could represent a deadly danger if it is too extensive, and this might be the case. Moreover, it could be observed that cellular damage continues further and becomes irrevocable after 6 h and MDA could not be detected. This may be due to the fact that the lipids were completely degraded and cells were no longer viable. Lipids from the cell membrane are the most prone to oxidation. In fact, lipid peroxidation biomarkers are used to screen the oxidative body balance [51]. At the same time, AOPP values are up to 30 times higher for bare nanoparticles in comparison to the functionalized ones.

Prof. Maged Younes, Chair of EFSA’s expert Panel on Food Additives and Flavourings (FAF), wrote of the decision: “Taking into account all available scientific studies and data, the Panel concluded that titanium dioxide can no longer be considered safe as a food additive. A critical element in reaching this conclusion is that we could not exclude genotoxicity concerns after consumption of titanium dioxide particles. After oral ingestion, the absorption of titanium dioxide particles is low, however they can accumulate in the body.”

The lack of clear regulations and controls explains that P25TiO₂NPs are still found in many of the commercialized sunscreens in the market. Some of them are coated to reduce the photoactivity of the anatase form, which is known to be responsible for tissue damage, but not enough studies were made on these coated forms. The anatase photoactivity could trigger the production of reactive oxygen species (ROS) generation, as it was stated before. The ROS are chemically reactive species containing oxygen, such as peroxides, superoxide, hydroxyl radical, and singlet oxygen. They are regularly produced in the biological milieu and counterbalanced by physiological antioxidant defense mechanisms. However, an abrupt increase of ROS may result in non-reversible damage to the skin cells. The effects of coated and uncoated P25TiO₂NPs need therefore to be studied, and articles on this topic present different conclusions. [11], [12], [13] Recent literature on this topic found that TiO₂NPs inhalation provokes serious genotoxicity and DNA damage [14], [15], [16], [17]. On the other hand, some studies in rats have reported no significant harm to genetic material [18], [19], [20], [21], [22].

The author thanks Marco Leona, Scientist-in-Charge of the Department of Scientific Research at the Metropolitan Museum of Art for conducting fluorescence spectrometry on Wheel of Fortune and a valuable discussion of the research, as well as Silvia Centeno, Research Scientist at the Metropolitan Museum of Art, who performed Raman analysis on the watercolors and also contributed her insight. The phenomenon of the phosphorescing lithopone was originally discovered during the author's fellowship in the Sherman Fairchild Center for the Conservation of Works on Paper, funded by the Andrew W. Mellon Foundation. The author thanks all her colleagues for their ideas and support during the research of this paper, and special thanks to Rachel Mustalish for her assistance in editing this work.

In a 2021, Chinese researchers examined the impact of E171 on lipid digestion and vitamin D3 bioaccessibility in a simulated human gastrointestinal tract model. They examined Vitamin D’s bioaccessibility, or the amount it was released in the gastrointestinal tract, becoming available for absorption, and found it “significantly decreased from 80% to 74%” with the addition of E171. In the experiment, E171 decreased lipid digestion dose-dependently. Researchers wrote: “The findings of this study enhance our understanding toward the potential impact of E171 on the nutritional attributes of foods for human digestion health.”  The study was published in the Journal of Agricultural and Food Chemistry,