Tattooing and use of permanent makeup (PMU) have dramatically increased over the last decade, with a concomitant increase in ink-related infections. Studies have shown evidence that commercial tattoo and PMU inks are frequently contaminated with pathogenic microorganisms. Considering that tattoo inks are placed into the dermal layer of the skin where anaerobic bacteria can thrive and cause infections in low-oxygen environments, the prevalence of anaerobic and aerobic bacteria should be assessed in tattoo and PMU inks. In this study, we tested 75 tattoo and PMU inks using the analytical methods described in the FDA Bacteriological Analytical Manual Chapter 23 for the detection of both aerobic and anaerobic bacterial contamination, followed by 16S rRNA gene sequencing for microbial identification. Of 75 ink samples, we found 26 contaminated samples with 34 bacterial isolates taxonomically classified into 14 genera and 22 species. Among the 34 bacterial isolates, 19 were identified as possibly pathogenic bacterial strains. Two species, namely Cutibacterium acnes (four strains) and Staphylococcus epidermidis (two strains) were isolated under anaerobic conditions. Two possibly pathogenic bacterial strains, Staphylococcus saprophyticus and C. acnes, were isolated together from the same ink samples (n = 2), indicating that tattoo and PMU inks can contain both aerobic (S. saprophyticus) and anaerobic bacteria (C. acnes). No significant association was found between sterility claims on the ink label and the absence of bacterial contamination. The results indicate that tattoo and PMU inks can also contain anaerobic bacteria.
OBJECTIVE: The rising popularity of tattooing and permanent makeup (PMU) has led to increased reports of ink-related infections. This study is the first to investigate the presence of both aerobic and anaerobic bacteria in commercial tattoo and PMU inks under aerobic and anaerobic conditions. Our findings reveal that unopened and sealed tattoo inks can harbor anaerobic bacteria, known to thrive in low-oxygen environments, such as the dermal layer of the skin, alongside aerobic bacteria. This suggests that contaminated tattoo inks could be a source of infection from both types of bacteria. The results emphasize the importance of monitoring these products for both aerobic and anaerobic bacteria, including possibly pathogenic microorganisms.

This paper presents the case of a 28-year-old with a history of chronic pelvic pain suspicious of endometriosis. She underwent a diagnostic laparoscopy and biopsy of lesions along the posterior cul-de-sac and right sidewall near the external iliac artery. Histopathological examination revealed no evidence of endometriosis but did comment on benign lymph node tissue with tattoo-like pigment. These findings correspond to the patient\'s tattoos located along the lower extremities. It is suspected there was cutaneous tattoo ink migration causing intra-abdominal lymphadenopathy, which visually mimicked endometriosis on diagnostic laparoscopy. Surgeons should become familiar with and recognize this phenomenon, as it can be misleading in the setting of endometriosis and diagnostic laparoscopy. Surgeons undertaking these cases must be able to identify and safely excise abnormal-appearing lesions in many different locations to prevent a missed or delayed diagnosis of endometriosis.

After tattoo application, inks remain in the skin, mostly in the dermal layer, and manufacturers use inks that have not been adequately evaluated for safety in tattoo production. In this study, the metal contents (Cd, Hg, Pb, and Cr) of tattoo inks available in the Turkish market were determined and the relationship between cell viability and inflammatory response of the detected metal levels was investigated. Nine tattoo inks (3 colors) from 3 different brands abbreviated as E, I, and W were examined. ICP-MS was used for element analysis. The viability of human keratinocyte cells was determined by the WST-1 assay following ink exposures at various dilutions. IL-18 levels were measured in cell culture supernatant by ELISA method following ink or metal (Cd, Cr, Hg, and Pb) exposures. The concentrations of trace elements were found in inks as follows: Cd, 0.0641-1.3857; Hg, 0.0204-0.2675; Pb, 0.8527-6.5981; Cr, 0.1731-45.3962 µg mL-1. It was observed that the levels of Pb and especially Cr in the samples exceeded the limit values. Tattoo inks reduced the cell viability in a dose- and color-dependent manner. IL-18 release was significantly increased in all groups except Cr and black ink of brand I treated cells (p < 0.05). Our results show that the metal contents of tattoo inks exceed Council of Europe Resolution values in some samples and some inks induce immune system activation (IL-18 secretion) and cytotoxic effects. It is thought that these findings may contribute to the toxic/adverse effects of tattoo inks commonly used.

UNASSIGNED: This study aimed to investigate the effects of commercial tattoo inks used in corneal tattooing on conjunctival microbiota.
UNASSIGNED: This prospective case control study consisted of 125 participants divided in the following three groups: 35 patients with corneal tattoos, 40 patients with corneal leukoma, and 50 healthy subjects. Corneal tattooing was performed in all the cases in this study using a tattoo pen machine and commercial tattoo ink. A total of 500 cultures were taken from 250 eyes of 125 individuals on chocolate and sheep blood agar. Bacteriological samples were taken from the inferior eyelid conjunctiva using a sterile cotton swab. Without any contact elsewhere, the swabs were smeared on bedside chocolate agars and 5% sheep blood agar.
UNASSIGNED: In tattooed eyes, bacterial growth was detected in 42.9% of the chocolate and sheep blood agar samples. In other healthy eyes of patients with corneal tattoos, 54.5% bacterial growth on chocolate agar and 57.1% on sheep blood agar were detected. No statistical difference was detected in the conjunctival microbiota of chocolate and sheep blood agar (p = 0.254, p = 0.134, respectively) in the tattooed eyes compared to the other eye of the individual. No statistically significant difference was found in terms of bacterial growth in tattooed, leukoma, or healthy eyes on chocolate and sheep blood agar (p = 0.408, p = 0.349). The growth rate of Staphylococcus epidermidis decreased by 33.3% (from 12 to 8) on chocolate agar in 35 tattooed eyes, and it decreased by 28.5% (from 14 to 10) on sheep blood agar, while gram-negative bacteria Brevundimonas diminuta, Acinetobacter lwoffii, and Psychrobacter faecalis were detected in three patients.
UNASSIGNED: Corneal tattooing using commercial dye does not affect conjunctival microbiota. In the past 3 years, 120 patients have been tattooed with commercial tattoo ink in Istanbul Medeniyet University Göztepe Training and Research Hospital. No complications related to infection were found in the 3-year follow-up. The gram-negative bacteria detected in the healthy control group and tattooed eyes were bacteria found on normal skin or in the respiratory tract. Although some gram-negative bacteria do not cause infection, careful eye examination, follow-up, and culture are required in suspicious cases.

Perception of skin color in the natural state, or with tattoo pigment installed in the dermis is highly complex and includes both optical requisites and eye and brain functions connected to advanced cognitive and psychological functions. Permanent makeup (PMU) is miniaturized tattooing on critical anatomical location aiming at fulfilment of a person\'s esthetic ideal. It is a special challenge since a satisfactory outcome depends on perfectionated technique and very critical selection of pigments. The eye can be compared to a digital camera, and the center of central vision in the brain to a computer and color integrator. Color perception is the end product of a complicated brain function. The background theory of color physics and perception is reviewed with emphasis on aspects related to changed skin color created by pigment installed in the dermis with the purpose to tattoo eyebrows, eyeliners, lips, and other locations. The delicate dosing of pigment relative to the natural skin tone is highlighted. The review is relevant for all aspects of PMU including medical tattooing, e.g., areola tattooing after breast cancer surgery. A detailed guidance to PMU artists and medical tattooists on selection and use of tattoo inks and pigments is provided. The guidance is also relevant to laser surgeons and technicians performing tattoo removal.

We outline constituents of tattoo and permanent make-up ink with regard to inflammatory tattoo reactions and population-based confounders. The comprehensive review of patch-tested tattoo patients between 1997 and 2022 shows that tattoo allergy cannot be reliably diagnosed via patch testing with today\'s knowledge. Weak penetration and slow haptenization of pigments, unavailability of pigments as test allergens and a lack of knowledge concerning relevant epitopes hamper the diagnosis of tattoo allergy. Patch testing p-phenylenediamine and disperse (textile) dyes is not able to close this gap. Sensitization to metals was associated with all types of tattoo complications, although often not clinically relevant for the tattoo reaction. Binders and industrial biocides are frequently missing on ink declarations and should be patch tested. The pigment carbon black (C.I. 77266) is no skin sensitizer. Patch tests with culprit inks were usually positive with cheap ink products for non-professional use or with professionally used inks in patients with eczematous reactions characterized by papules and infiltration. Tape stripping before patch testing and patch test readings on Day 8 or 10 may improve the diagnostic quality. The meaningfulness of the categorical EU-wide ban of Pigment Green 7 and Pigment Blue 15:3 is not substantiated by the presented data.

Tattoo inks are comprised of different combinations of bioactive chemicals with combined biological effects that are insufficiently explored. Tattoos have been associated with oxidative stress; however, a recent N-of-1 study suggested that blue tattoos may be associated with suppressed local skin oxidative stress. The present study aimed to explore the attributes of the blue tattoo ink (BTI) that may explain its possible effects on redox homeostasis, namely the catalase (CAT) and superoxide dismutase (SOD)-mimetic properties that have been reported for copper(II) phthalocyanine (CuPC)-the main BTI constituent. Intenze™ Persian blue (PB) BTI has been used in the experiment. CAT and SOD-mimetic properties of PB and its pigment-enriched fractions were analyzed using the carbonato-cobaltate (III) formation-derived H2O2 dissociation and 1,2,3-trihydroxybenzene autoxidation rate assays utilizing simple buffers and biochemical matrix of normal skin tissue as chemical reaction environments. CuPC-based tattoo ink PB and both its blue and white pigment-enriched fractions demonstrate CAT and SOD-mimetic properties in vitro with effect sizes demonstrating a substantial dependence on the biochemical environment. PB constituents act as inhibitors of CAT but potentiate its activity in the biochemical matrix of the skin. CuPC-based BTI can mimic antioxidant enzymes, however chemical constituents other than CuPC (e.g. the photoreactive TiO2) seem to be at least partially responsible for the BTI redox-modulating properties.

BACKGROUND: According to the REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals) restriction, tattoo and permanent make-up (PMU) inks placed on the European Union market after January 4, 2022, shall not contain methylisothiazolinone, benzisothiazolinone (BIT), octylisothiazolinone (OIT), or other skin sensitizers in concentrations of 10 mg/kg or higher and phenoxyethanol (PE) or other eye irritants or damaging substances in concentrations of 100 mg/kg or higher. In addition, preservatives and other substances enlisted in Annex II to Cosmetic Product Regulation shall not be present in concentrations of 0.5 mg/kg or higher.
OBJECTIVE: This study aims to quantify 14 preservatives in 99 tattoo and 39 PMU inks from the Italian market and presents a comparison with concentration limits set by the REACH restriction.
METHODS: Inks were analysed by applying validated analytical methods based on liquid chromatography techniques.
RESULTS: About 24.0%, 15.2% and 1.5% of the overall samples contained BIT, PE and OIT, respectively, at concentrations exceeding REACH concentration limits. The number of noncompliant tattoo inks (49.5%) would be significantly greater than that of the PMU inks (17.9%).
CONCLUSIONS: About 40.6% of the samples would be noncompliant with the restriction for the presence of preservatives above the permitted level. Additional concentration limits will apply to skin sensitizing preservatives for proper labelling of inks under CLP (Classification, Labelling and Packaging) Regulation.

Tattooing is an ancient practice and its popularity has been increasing in the recent years. After tattooing, complications may occur related to compose tattoo inks. In this study, the phototoxicity potential of the blue, red and black colors of the most commonly used three different commercially-available tattoo ink brands have been examined by performing in vitro 3T3-neutral red uptake (NRU) phototoxicity test.
In the study, the phototoxicity of serial diluted concentrations of tattoo inks were evaluated with in vitro 3T3-NRU phototoxicity test method according to OECD guide 432. The data obtained from the NRU test result were uploaded to Phototox software (version 2.0) and the phototoxicity potentials of tattoo inks were determined via the calculation of the mean photo effect (MPE) and photo irritation factor (PIF) values.
The red, black and blue colors of three different commercially available tattoo inks did not cause a cytotoxic activity on BALB/c 3T3 cells with 3T3-NRU test. The IC50 values could not be determined +ultraviolet (UV) and -UV conditions. PIF values could not be calculated and MPE values were <0.1, which predicts the absence of phototoxic effect for all of the tested tattoo inks.
All tested inks were evaluated as non-phototoxic according to the results of MPE values calculated using Phototox software. However, test results should be verified by other phototoxicity test methods to obtain a comprehensive evaluation of phototoxic complications of different tattoo inks.

A variety of side effects following the tattooing of the skin were reported over the years. Analytical studies showed that some tattoo inks contain harmful compounds.
We presented six patient cases with cutaneous malignancies in tattooed skin and performed an extensive literature research.
Two patients with black ink tattoos that were diagnosed with malignant melanoma raises the number of described cases to 36 patients. One of the patients developed an immunologic reaction limited to the tattoo area after treatment with a targeted immune therapy. In the other patient, the malignancy (malignant melanoma) was fatal. Basal cell carcinoma was seen in four patients with tattoos containing varying ink colors (black, green, red). This increased the number of described patient cases to 18. Although some ink components and their cleavage products have carcinogenic properties, epidemiological evidence for a causative correlation fails. Further epidemiologic studies on tattoos and malignancies, as well as on the appearance of naevi in tattoos, are necessary. Determining the type of mutation might be helpful to separate sun-induced tumors from skin cancers due to other pathogenic mechanisms.