Preparation of Autoclavable and Injectable Silk Fibroin Cryogels for Tissue Engineering Applications.

A cryogel is a supermacroporous gel network that is generated at subzero temperatures by polymerizing monomers or gelating polymeric precursors. Since cryogels possess inherent characteristics such as interconnected macroporous structures, excellent mechanical properties, and high resistance to autoclave sterilization, they are highly desirable for tissue engineering and regenerative medicine. Silk fibroin, a natural protein obtained from Bombyx mori silkworms, is an excellent raw material for cryogel preparation. The aim of this study is to establish a controlled method for preparing silk fibroin cryogels with suitable properties for application as tissue engineering scaffolds. Using a dual crosslinking strategy consisting of low-temperature radical polymerization coupled with methanol-induced conformational transformation, porous cryogels are prepared. The cryogels display many unique characteristics, such as an interconnected macroporous structure, a high water absorption capacity, water-triggered shape memory, syringe injectability, and strong resilience to autoclave sterilization. Furthermore, the cryogels demonstrate excellent biocompatibility and cell affinity, facilitating cell adhesion, migration, and proliferation. The interconnected supermacroporous architecture resembling the native extracellular matrix, together with their unique physical properties and autoclaving stability, suggests that cryogels are promising candidate scaffolds for tissue engineering and cell therapy.

[Research advances of biomedical polymeric microneedles for non-transdermal local drug delivery].

Microneedles have emerged as the new class of local drug delivery system that has broad potential for development. Considering that the microneedles can penetrate tissue barriers quickly, and provide localized and targeted drug delivery, their applications have gradually expanded to non-transdermal drug delivery recently, which are capable of providing rapid and effective treatment for injuries and diseases of organs or tissues. However, a literature search revealed that there is a lack of summaries of the latest developments in non-transdermal drug delivery research by using biomedical polymeric microneedles. The review first described the materials and fabrication methods for the polymeric microneedles, and then reviewed a representative application of microneedles for non-transdermal drug delivery, with the primary focus being on treating and repairing the tissues or organs such as oral cavity, ocular tissues, blood vessels and heart. At the end of the article, the opportunities and challenges associated with microneedles for non-transdermal drug delivery were discussed, along with its future development, in order to provide reference for researchers in the relevant field.

Trophic behaviors of PFOA and its alternatives perfluoroalkyl ether carboxylic acids (PFECAs) in a coastal food web.

With the increasing restrictions and concerns about legacy poly- and perfluoroalkyl substances (PFAS), the production and usage of alternatives, i.e., perfluoroalkyl ether carboxylic acids (PFECAs), have risen recently. However, there is a knowledge gap regarding the bioaccumulation and trophic behaviors of emerging PFECAs in coastal ecosystems. The bioaccumulation and trophodynamics of perfluorooctanoic acid (PFOA) and its substitutes (PFECAs) were investigated in Laizhou Bay, which is located downstream of a fluorochemical industrial park in China. Hexafluoropropylene oxide trimer acid (HFPO-TrA), perfluoro-2-methoxyacetic acid (PFMOAA) and PFOA constituted the dominant compounds in the ecosystem of Laizhou Bay. PFMOAA was dominant in invertebrates, whereas the long-chain PFECAs preferred to accumulate in fishes. The PFAS concentrations in carnivorous invertebrates were higher than those in filter-feeding species. Considering migration behaviors, the ∑PFAS concentrations followed the order oceanodromous fish < diadromous fish < non-migratory fish. The trophic magnification factors (TMFs) of long-chain PFECAs (HFPO-TrA, HFPO-TeA and PFO5DoA) were >1, suggesting trophic magnification potential, while biodilution for short-chain PFECAs (PFMOAA) was observed. The intake of PFOA in seafood may constitute a great threat to human health. More attention should be given to the impact of emerging hazardous PFAS on organisms for the health of ecosystems and human beings.

Emerging and Legacy Perfluoroalkyl Substances in Breastfed Chinese Infants: Renal Clearance, Body Burden, and Implications.

BACKGROUND: Human breast milk is a primary route of exposure to perfluoroalkyl substances (PFAS) in infants. To understand the associated risks, the occurrence of PFAS in human milk and the toxicokinetics of PFAS in infants need to be addressed. OBJECTIVES: We determined levels of emerging and legacy PFAS in human milk and urine samples from Chinese breastfed infants, estimated renal clearance, and predicted infant serum PFAS levels. METHODS: In total, human milk samples were collected from 1,151 lactating mothers in 21 cities in China. In addition, 80 paired infant cord blood and urine samples were obtained from two cities. Nine emerging PFAS and 13 legacy PFAS were analyzed in the samples using ultra high-performance liquid chromatography tandem mass spectrometry. Renal clearance rates (CLrenals) of PFAS were estimated in the paired samples. PFAS serum concentrations in infants (<1 year of age) were predicted using a first-order pharmacokinetic model. RESULTS: All nine emerging PFAS were detected in human milk, with the detection rates of 6:2 Cl-PFESA, PFMOAA, and PFO5DoDA all exceeding 70%. The level of 6:2 Cl-PFESA in human milk (median concentration=13.6 ng/L) ranked third after PFOA (336 ng/L) and PFOS (49.7 ng/L). The estimated daily intake (EDI) values of PFOA and PFOS exceeded the reference dose (RfD) of 20 ng/kg BW per day recommended by the U.S. Environmental Protection Agency in 78% and 17% of breastfed infant samples, respectively. 6:2 Cl-PFESA had the lowest infant CLrenal (0.009mL/kg BW per day), corresponding to the longest estimated half-life of 49 y. The average half-lives of PFMOAA, PFO2HxA, and PFO3OA were 0.221, 0.075, and 0.304 y, respectively. The CLrenals of PFOA, PFNA, and PFDA were slower in infants than in adults. CONCLUSIONS: Our results demonstrate the widespread occurrence of emerging PFAS in human milk in China. The relatively high EDIs and half-lives of emerging PFAS suggest potential health risks of postnatal exposure in newborns. https://doi.org/10.1289/EHP11403.

Unveiling Distribution of Per- and Polyfluoroalkyl Substances in Matched Placenta-Serum Tetrads: Novel Implications for Birth Outcome Mediated by Placental Vascular Disruption.

The placenta is pivotal for fetal development and maternal-fetal transfer of many substances, including per- and polyfluoroalkyl substances (PFASs). However, the intraplacental distribution of PFASs and their effects on placental vascular function remain unclear. In this study, 302 tetrads of matched subchorionic placenta (fetal-side), parabasal placenta (maternal-side), cord serum, and maternal serum samples were collected from Guangzhou, China. Eighteen emerging and legacy PFASs and five placental vascular biomarkers were measured. Results showed that higher levels of perfluorooctanoic (PFOA), perfluorooctane sulfonic acid (PFOS), and chlorinated polyfluorinated ether sulfonic acids (Cl-PFESAs) were detected in subchorionic placenta compared to parabasal placenta. There were significant associations of PFASs in the subchorionic placenta, but not in the serum, with placental vascular biomarkers (up to 32.5%) and lower birth size. Birth weight was negatively associated with PFOA (ß: -103.8, 95% CI: -186.3 and -21.32) and 6:2 Cl-PFESA (ß: -80.04, 95% CI: -139.5 and -20.61), primarily in subchorionic placenta. Mediation effects of altered placental angiopoietin-2 and vascular endothelial growth factor receptor-2 were evidenced on associations of adverse birth outcomes with intraplacental PFOS and 8:2 Cl-PFESA, explaining 9.5%-32.5% of the total effect. To the best of our knowledge, this study is the first to report on differential intraplacental distribution of PFASs and placental vascular effects mediating adverse birth outcomes and provides novel insights into the placental plate-specific measurement in PFAS-associated health risk assessment.

Comprehensive profiles of per- and polyfluoroalkyl substances in Chinese and African municipal wastewater treatment plants: New implications for removal efficiency.

Municipal wastewater treatment plants (WWTPs) can reflect the pollution status of per- and polyfluoroalkyl substances (PFASs) pollution. Here, matched influent, effluent, and sludge samples were collected from 58 municipal WWTPs in China, South Sudan, Tanzania, and Kenya. Target and suspect screening of PFASs was performed to explore their profiles in WWTPs and assess removal efficiency and environmental emissions. In total, 155 and 58 PFASs were identified in WWTPs in China and Africa, respectively; 146 and 126 PFASs were identified in wastewater and sludge, respectively. Novel compounds belonging to per- and polyfluoroalkyl ether carboxylic acids (PFECAs) and sulfonic acids (PFESAs), hydrogen-substituted polyfluorocarboxylic acids (H-PFCAs), and perfluoroalkyl sulfonamides (PFSMs) accounted for a considerable proportion of total PFASs (ΣPFASs) in Chinese WWTPs and were also widely detected in African samples. In China, estimated national emissions of ΣPFASs in WWTPs exceeded 16.8 t in 2015, with >60 % originating from emerging PFASs. Notably, current treatment processes are not effective at removing PFASs, with 35 of the 54 WWTPs showing emissions higher than mass loads. PFAS removal was also structure dependent. Based on machine learning models, we found that molecular descriptors (e.g., LogP and molecular weight) may affect adsorption behavior by increasing hydrophobicity, while other factors (e.g., polar surface area and molar refractivity) may play critical roles in PFAS removal and provide novel insights into PFAS pollution control. In conclusion, this study comprehensively screened PFASs in municipal WWTPs and determined the drivers affecting PFAS behavior in WWTPs based on machine learning models.

Comparative Hepatotoxicity of a Novel Perfluoroalkyl Ether Sulfonic Acid, Nafion Byproduct 2 (H-PFMO2OSA), and Legacy Perfluorooctane Sulfonate (PFOS) in Adult Male Mice.

Nafion byproduct 2 (H-PFMO2OSA) has been detected in the environment, but little is known about its toxicities. To compare the hepatotoxicity of H-PFMO2OSA with legacy perfluorooctane sulfonate (PFOS), male adult mice were exposed to 0.2, 1, or 5 mg/kg/d of each chemical for 28 days. Results showed that, although H-PFMO2OSA liver and serum concentrations were lower than those of PFOS, the relative liver weight in the H-PFMO2OSA groups was significantly higher than that in the corresponding PFOS groups. In addition, the increase in alanine transaminase and aspartate aminotransferase activity was greater in the H-PFMO2OSA groups than in the PFOS groups. Reduced glutathione (GSH) content and glutathione reductase activity in the liver increased in the 1 and 5 mg/kg/d H-PFMO2OSA groups and in the 5 mg/kg/d PFOS group. Liver quantitative proteome analysis demonstrated that, similar to PFOS, H-PFMO2OSA caused lipid metabolism disorder, and most lipid metabolism-related differentially expressed proteins (DEPs) were controlled by peroxisome proliferator-activated receptor alpha (PPARα). Additionally, KEGG enrichment analysis highlighted changes in the GSH metabolism pathway after PFOS and H-PFMO2OSA exposure. Then, there were eight DEPs involved in the GSH metabolism pathway that mostly were upregulated after exposure to H-PFMO2OSA but not after exposure to PFOS. In conclusion, H-PFMO2OSA induced higher levels of liver damage and more serious GSH metabolism dysregulation compared to PFOS.

Nontargeted Identification and Temporal Trends of Per- and Polyfluoroalkyl Substances in a Fluorochemical Industrial Zone and Adjacent Taihu Lake.

Various per- and polyfluoroalkyl substances (PFASs) remain undiscovered and unexplored in the environment. The goals of this study were to discover new species of PFASs in effluent and surface waters from a fluorochemical industrial zone, and to assess their concentration, distribution, and temporal trends in the adjacent natural environment. In total, 83 emerging PFASs from 14 classes were identified, 22 of which were reported for the first time. Authentic standards were synthesized for 13 per- and polyfluoroalkyl ether carboxylic acids (PFECAs), thereby greatly expanding the scope of PFAS-targeted monitoring. The newly identified compounds accounted for 27%-95% of the total PFAS concentrations. Of note, a novel diether carboxylic acid, 2-[2-(trifluoromethoxy)hexafluoropropoxy]tetrafluoropropanoic acid (C7 HFPO-TA) was detected at an extremely high concentration in the fluorochemical zone effluent (447 000 ng/L) and at a median concentration in the fluorochemical zone surface water (670 ng/L), with detectable levels also found in the natural environment, that is, Wangyu River (23 ng/L) and Taihu Lake (5.6 ng/L). The distinct geographic distribution of C7 HFPO-TA suggests transport from the industrial point source to Taihu Lake via the Wangyu River. The concentration of C7 HFPO-TA in Taihu Lake, along with that of many other emerging PFASs, continued to grow in three sampling campaigns from 2016 to 2021. Considering the environmental persistence and toxicity of structurally similar PFECAs (e.g., HFPO-DA), studies on C7 HFPO-TA are urgently needed.

Transcriptome analysis of 3D primary mouse liver spheroids shows that long-term exposure to hexafluoropropylene oxide trimer acid disrupts hepatic bile acid metabolism.

Hexafluoropropylene oxide trimer acid (HFPO-TA), an alternative to perfluorooctanoic acid (PFOA), has been detected in various environmental and human matrices. However, information regarding its toxicity remains limited. Here, we established a three-dimensional (3D) primary mouse liver spheroid model to compare the hepatotoxicity of HFPO-TA and PFOA. The 3D spheroids were repeatedly exposed to 25-, 50-, or 100-µM HFPO-TA and PFOA for 28 d. Compared with the PFOA groups, the HFPO-TA groups showed higher bioaccumulation potential, higher lactate dehydrogenase (LDH) leakage, and lower adenosine triphosphate (ATP), albumin, and urea secretion. Transcriptome analysis identified 1603 and 772 differentially expressed genes in the 100-µM HFPO-TA- and PFOA-treated groups, respectively. Bioinformatics analysis indicated that cholesterol metabolism, bile acid metabolism, and inflammatory response were significantly altered. Exposure to 100-µM HFPO-TA increased triglyceride content but decreased total cholesterol content, while no changes were observed in the 100-µM PFOA-treated group. Total bile acids in the re-polarized 3D spheroids increased significantly after 100-µM HFPO-TA and PFOA treatment, which did not affect bile acid synthesis but inhibited the expression levels of Bsep and Mrp2 related to bile acid transport. Thus, HFPO-TA exhibited more serious hepatotoxicity than PFOA in 3D primary liver spheroids and may not be a safe alternative.

Exposure to GenX and Its Novel Analogs Disrupts Hepatic Bile Acid Metabolism in Male Mice.

Due to its wide usage and recent detection in environmental matrices, hexafluoropropylene oxide dimer acid (HFPO-DA, commercial name GenX) has attracted considerable attention. Here, we explored and compared the toxicity of GenX and its novel analogs with that of perfluorooctanoic acid (PFOA) to provide guidance on the structural design and optimization of novel alternatives to poly- and perfluoroalkyl substances (PFASs). Adult male BALB/c mice were continuously exposed to PFOA, GenX, perfluoro-2-methyl-3,6-dioxo-heptanoic acid (PFMO2HpA), and perfluoro-2-methyl-3,6,8-trioxo-nonanoic acid (PFMO3NA; 0, 0.4, 2, or 10 mg/kg/d) via oral gavage for 28 days. The PFOA, GenX, and PFMO3NA treatment groups showed an increase in relative liver weight, and bile acid metabolism was the most significantly affected pathway in all treatment groups, as shown via weighted gene coexpression network analysis. The highest total bile acid levels were observed in the 2 and 10 mg/kg/d PFMO3NA groups. The ratios of primary bile acids to all bile acids increased in the high-dose groups, while the ratios of secondary bile acids showed a downward trend. Thus, bile acid metabolism disorder may be a prominent adverse effect induced by exposure to GenX, its analogs, and PFOA. Results also showed that the hepatotoxicity of PFMO2HpA was lower than that of GenX, whereas the hepatotoxicity of PFMO3NA was stronger, suggesting that PFMO2HpA may be a potential alternative to GenX.

First Report on the Bioaccumulation and Trophic Transfer of Perfluoroalkyl Ether Carboxylic Acids in Estuarine Food Web.

As novel alternatives to legacy poly- and perfluoroalkyl substances (PFAS), perfluoroalkyl ether carboxylic acids (PFECAs) have been widely detected in the environment; however, there is limited information and knowledge regarding their bioaccumulation and trophic transfer behavior along the food chain. This research presents the first known published data on the bioaccumulation and trophic transfer characteristics of PFECAs in a source-impacted estuary. Elevated PFECA concentrations were observed in organisms (for instance, conch, with perfluoro-2-methoxyacetic acid (PFMOAA) concentration reaches up to 16 700 ng/g dry weight (dw)), indicating exposure risks to the consumers. Conch can be acted as a potential environmental bioindicator of PFMOAA. PFMOAA, hexafluoropropylene oxide trimer acid (HFPO-TrA) and PFOA were predominant detected in biotas. On the basis of trophic magnification factors (TMFs), PFECAs with ≥6 perfluorinated carbons (HFPO-TrA, hexafluoropropylene oxide tetramer acid (HFPO-TeA) and perfluoro (3, 5, 7, 9, 11-pentaoxadodecanoic) acid (PFO5DoA)) could be biomagnified along the food chain (TMF > 1), while PFMOAA with the least perfluorinated carbons undergone biodilution (TMF < 1). As seafood is an important dietary source of protein to human, there is a potential health risk related to the consuming polluted aquatic products.

Chronic exposure to PFO4DA and PFO5DoDA, two perfluoroalkyl ether carboxylic acids (PFECAs), suppresses hepatic stress signals and disturbs glucose and lipid metabolism in male mice.

Perfluoroalkyl ether carboxylic acids (PFECAs), including PFO4DA and PFO5DoDA, have been found in both surface water and volunteer blood samples from polluted regions. However, little knowledge is available on their potential bioaccumulation and health risk. In the present study, the half-lives of PFO4DA and PFO5DoDA in male mouse serum were 24 h and nearly 43 d, respectively, indicating markedly increased difficulty in eliminating PFO5DoDA from the body. After 140 d daily exposure both PFO4DA and PFO5DoDA (10 µg/kg/d) increased body weight. Hepatomegaly was the most sensitive phenomenon after exposure treatment, with occurrence even in the 2 µg/kg/d exposure groups. RNA-seq analysis supported a similar but stronger effect of PFO5DoDA compared with PFO4DA. A wide array of genes involved in stimulus sensing and response were suppressed. In addition to weight gain, hyperglycemia was also observed after treatment. Increased glucose and decreased pyruvate and lactate levels in the liver supported a reduction in glycolysis, consistent with the reduction in the key regulator Pfkfb3. In conclusion, chronic PFO4DA and PFO5DoDA exposure suppressed stress signals and disturbed glucose and lipid metabolism in the liver. The longer serum half-life and stronger hepatic bioaccumulation of PFO5DoDA, at least partially, contributed to its stronger hepatotoxicity than that of PFO4DA.

Novel Perfluoroalkyl Ether Carboxylic Acids (PFECAs) and Sulfonic Acids (PFESAs): Occurrence and Association with Serum Biochemical Parameters in Residents Living Near a Fluorochemical Plant in China.

Although perfluoroalkyl ether carboxylic (PFECAs) and sulfonic acids (PFESAs) have been widely detected in environmental matrices, their occurrence in humans and impact on human health remains insufficiently understood. Here, we report on 13 PFECAs and PFESAs in 977 sera samples collected from residents living near a fluorochemical plant in Shandong, China. The sum concentration of these emerging PFECAs accounted for 13% of the total PFASs in the serum of the participants, with the frequent detection of several PFECAs (>95%) (PFMOAA, PFO4DA, and PFO5DoDA at median concentrations of 12.91, 0.142, and 0.987 ng/mL, respectively) and PFESAs (98.7%) (Nafion byproduct 2 at a median concentration of 0.097 ng/mL). Serum PFMOAA, PFO5DoDA, and 6:2 Cl-PFESA levels were significantly higher in males than in females. Positive relationships were observed between age and PFMOAA, 6:2 Cl-PFESA, and H-PFMO2OSA levels, whereas HFPO-TA and PFO5DoDA serum concentrations in the 0-40-year age group were lower than that in the >40-year age group. Furthermore, multivariate linear regression models and sensitivity analyses showed positive associations among PFO5DoDA levels, elevated lipid parameters (cholesterol, low-density lipoprotein cholesterol, and triglycerides), liver function markers (albumin levels and alanine transaminase, aspartate aminotransferase, and glutamyl transpeptidase activities), and uric acid levels. Thus, our results suggest potential health risks from exposure to novel PFESAs and PFECAs (especially PFO5DoDA).

Association between exposure to per- and polyfluoroalkyl substances and blood glucose in pregnant women.

Increasing evidence shows that exposure to per- and polyfluoroalkyl substances (PFASs), common and persistent anthropogenic pollutants, may affect glucose homeostasis. However, data is limited for pregnant women, and it is less clear how novel fluorinated pollutants affect glucose homeostasis during pregnancy. Our goal was to investigate the relationships of exposure to13 PFASs including some novel fluorinated pollutants with blood glucose in 874 Chinese pregnant women from the general population. We measured blood glucose following an oral glucose tolerance test (OGTT) in the second trimester and quantified PFAS concentrations in umbilical cord blood. The associations of PFAS exposure with fasting, 1-h OGTT and 2-h OGTT glucose were examined using the general linear regression model. For every doubling of perfluorobutane sulfonate (PFBS), a short-chain PFAS, the 1-h and 2-h OGTT blood glucose increased 0.09 mmol/L (ß = 0.09, 95%CI: 0.02, 0.17) and 0.07 mmol/L (ß = 0.07, 95%CI: 0.01, 0.13), respectively. In addition, perfluoroheptanoate (PFHpA), an alternative of perfluorooctanoate (PFOA), was positively associated with fasting glucose (ß = 0.07, 95%CI: 0.02, 0.13; high vs low). Higher levels of PFAS exposure were related to increased blood glucose in pregnant women, indicating PFAS exposure may impair glucose homeostasis during pregnancy.

Per- and polyfluoroalkyl substances (PFASs) in blood of captive Siberian tigers in China: Occurrence and associations with biochemical parameters.

Per- and polyfluoroalkyl substances (PFASs) have been ubiquitously detected in the environment and marine animals. However, little is known about these substances and their associations with health parameters in wild terrestrial mammals. In this study, we determined PFAS levels and distribution in the blood of captive Siberian tigers in Harbin, China, and evaluated potential exposure pathways by daily intake. In addition, for the first time, we explored the associations between serum PFAS concentrations and clinical parameters. Results showed that perfluorooctanoate (PFOA) was the dominant PFAS compound in blood (accounting for 64%), followed by perfluorooctanesulfonate (PFOS). In addition, 6:2 chlorinated polyfluorinated ether sulfonate (6:2 Cl-PFESA) concentrations were also detected in blood and dietary food. Furthermore, significant positive age relationships were observed for levels of perfluoroheptanoate (PFHpA), PFOA, PFOS, and 6:2 Cl-PFESA in the blood of female tigers. Results showed that PFOA and PFOS in dietary food accounted for over 70% of total daily intake of PFASs, indicating that meat consumption is a predominant exposure pathway in tigers. We also found positive associations between higher exposure to PFASs (including PFOA, PFOS, and 6:2 Cl-PFESA) and elevated serum levels of alanine transaminase (ALT), a marker of liver damage. Thus, comprehensive health assessments of PFAS burdens in wildlife are needed.

Perfluoropolyether carboxylic acids (novel alternatives to PFOA) impair zebrafish posterior swim bladder development via thyroid hormone disruption.

Perfluoropolyether carboxylic acids (PFECAs, CF3(OCF2)nCOO-, n = 2-5) are novel alternatives to perfluorooctanoic acid (PFOA) and are widely used in industrial production. However, although they have been detected in surface water and human blood, their toxicities on aquatic organisms remain unknown. We used zebrafish embryos to compare the developmental toxicities of various PFECAs (e.g., perfluoro (3,5,7-trioxaoctanoic) acid (PFO3OA), perfluoro (3,5,7,9-tetraoxadecanoic) acid (PFO4DA), and perfluoro (3,5,7,9,11-pentaoxadodecanoic) acid (PFO5DoDA)) with that of PFOA and to further reveal the key events related to toxicity caused by these chemicals. Results showed that, based on half maximal effective concentrations (EC50), toxicity increased in the order: PFO5DoDA > PFO4DA > PFOA > PFO3OA, with uninflated posterior swim bladders the most frequently observed malformation. Similar to PFOA, PFECA exposure significantly lowered thyroid hormone (TH) levels (e.g., T3 (3,5,3'-L-triiodothyronine) and T4 (L-thyroxine)) in the whole body of larvae at 5 d post-fertilization following disrupted TH metabolism. In addition, the transcription of UDP glucuronosyltransferase 1 family a, b (ugt1ab), a gene related to TH metabolism, increased dose-dependently. Exogeneous T3 or T4 supplementation partly rescued PFECA-induced posterior swim bladder malformation. Our results further suggested that PFECAs primarily damaged the swim bladder mesothelium during early development. This study is the first to report on novel emerging PFECAs as thyroid disruptors causing swim bladder malformation. Furthermore, given that PFECA toxicity increased with backbone OCF2 moieties, they may not be safer alternatives to PFOA.

Comparative Hepatotoxicity of Novel PFOA Alternatives (Perfluoropolyether Carboxylic Acids) on Male Mice.

As novel alternatives to perfluorooctanoic acid (PFOA), perfluoropolyether carboxylic acids (multiether PFECAs, CF3(OCF2) nCOO-, n = 2-4) have been detected in various environmental matrices; however, public information regarding their toxicities remains unavailable. To compare the hepatotoxicity of multiether PFECAs (e.g., PFO2HxA, PFO3OA, and PFO4DA) with PFOA, male mice were exposed to 0.4, 2, or 10 mg/kg/d of each chemical for 28 d, respectively. Results demonstrated that PFO2HxA and PFO3OA exposure did not induce marked increases in relative liver weight; whereas 2 and 10 mg/kg/d of PFO4DA significantly increased relative liver weight. Furthermore, PFO2HxA and PFO3OA demonstrated almost no accumulation in the liver or serum; whereas PFO4DA was accumulated but with weaker potential than PFOA. Exposure to 10 mg/kg/d of PFO4DA led to 198 differentially expressed liver genes (56 down-regulated, 142 up-regulated), with bioinformatics analysis highlighting the urea cycle disorder. Like PFOA, 10 mg/kg/d of PFO4DA decreased the urea cycle-related enzyme protein levels (e.g., carbamoyl phosphate synthetase 1) and serum ammonia content in a dose-dependent manner. Both PFOA and PFO4DA treatment (highest concentration) caused a decrease in glutamate content and increase in both glutamine synthetase activity and aquaporin protein levels in the brain. Thus, we concluded that PFO4DA caused hepatotoxicity, as indicated by hepatomegaly and karyolysis, though to a lesser degree than PFOA, and induced urea cycle disorder, which may contribute to the observed toxic effects.