Exploring the osteogenic potential of semisynthetic triterpenes from Combretum leprosum: An in vitro and in silico study.

Combretum leprosum Mart. is a plant of the Combretaceae family, widely distributed in the Northeast region of Brazil, popularly used as an anti-inflammatory agent, and rich in triterpenes. This study evaluated in vitro and in silico potential osteogenic of two semisynthetic triterpenes (CL-P2 and CL-P2A) obtained from the pentacyclic triterpene 3ß,6ß,16ß-trihydroxylup-20(29)-ene (CL-1) isolated from C. leprosum. Assays were carried out in cultured murine osteoblasts (OFCOL II), first investigating the possible toxicity of the compounds on these cells through viability assays (MTT). Cell proliferation and activation were investigated by immunohistochemical evaluation of Ki-67, bone alkaline phosphatase (ALP) activity, and mineralization test by Von Kossa. Molecular docking analysis was performed to predict the binding affinity of CL-P2 and CL-P2A to target proteins involved in the regulation of osteogenesis, including: bone morphogenetic protein 2 (BMP-2), proteins related to Wingless-related integration (WNT) pathway (Low-density lipoprotein receptor-related protein 6-LRP6 and sclerostin-SOST), and receptor activator of nuclear factor (NF)-kB-ligand (RANK-L). Next, Western Blot and immunofluorescence investigated BMP-2, WNT, RANK-L, and OPG protein expressions in cultured murine osteoblasts (OFCOL II). None of the CL-P2 and CL-P2A concentrations were toxic to osteoblasts. Increased cell proliferation, ALP activity, and bone mineralization were observed. Molecular docking assays demonstrated interactions with BMP-2, LRP6, SOST, and RANK-L/OPG. There was observed increased expression of BMP-2, WNT, and RANK-L/OPG proteins. These results suggest, for the first time, the osteogenic potential of CL-P2 and CL-P2A.

Cytometry in the Short-Wave Infrared.

Cytometry plays a crucial role in characterizing cell properties, but its restricted optical window (400-850 nm) limits the number of stained fluorophores that can be detected simultaneously and hampers the study and utilization of short-wave infrared (SWIR; 900-1700 nm) fluorophores in cells. Here we introduce two SWIR-based methods to address these limitations: SWIR flow cytometry and SWIR image cytometry. We develop a quantification protocol for deducing cellular fluorophore mass. Both systems achieve a limit of detection of ∼0.1 fg cell-1 within a 30 min experimental time frame, using individualized, high-purity (6,5) single-wall carbon nanotubes as a model fluorophore and macrophage-like RAW264.7 as a model cell line. This high-sensitivity feature reveals that low-dose (6,5) serves as an antioxidant, and cell morphology and oxidative stress dose-dependently correlate with (6,5) uptake. Our SWIR cytometry holds immediate applicability for existing SWIR fluorophores and offers a solution to the issue of spectral overlapping in conventional cytometry.

Resmetirom for treatment of MASH.

Resmetirom is an oral selective THR-ß agonist conditionally approved for the treatment of patients with noncirrhotic MASH with moderate to advanced fibrosis. Resmetirom restores mitochondrial and hepatic metabolic function; reduces atherogenic lipids; improves hepatic steatosis, inflammation, and fibrosis; and has no significant effect on THR-α. To view this Bench to Bedside, open or download the PDF.

Fibrin Network and Platelets Densities in Platelet-Rich Fibrin (PRF) Membranes Produced from Plastic Tubes Without Additives: A New Approach to PRF Clinical Use.

Background/Purpose: The present study aimed to investigate plastic tubes without additives as alternatives to glass and silica-coated plastic tubes, in the production of PRF membranes. Materials and Methods: Nine blood samples were collected from eight volunteers (n = 8) separated into three groups, according to tube material: glass, silica-coated plastic, and plastic without additives. In each group, the samples were centrifuged using different relative centrifugation forces: L-PRF (700 g/12 min), A-PRF (200 g/14 min), and A-PRF + (200 g/8 min). The generated membranes were evaluated by histomorphometry, considering the fibrin network, platelet aggregates, and cellular morphology, by light microscopy. The ultrastructural cellular morphology integrity was evaluated by transmission electron microscopy. Results: The L-PRF (p < 0.019) and A-PRF (p < 0.001) membranes showed a significantly lower fibrin network density in plastic tubes without additives compared to glass and silica-coated plastic tubes. Plastic tubes without additives revealed a significantly higher platelet percentage, regardless of the protocol (p < 0.005). In all groups, TEM analysis showed preserved normal morphological ultrastructure, maintaining the integrity of cellular components. Conclusion: Plastic tubes without additives offer a viable alternative for producing PRF membranes. They exhibited a higher platelet density and demonstrated fibrin network and cellular morphology similar to those of glass and silica-coated plastic tubes, irrespective of the centrifugation protocol.

Reviving forgotten techniques: Vaginal salpingectomy in public health system, a retrospective cohort study.

OBJECTIVE: To evaluate the viability of the vaginal route as an alternative to laparoscopy for patients seeking permanent surgical contraception. METHODS: We conducted a retrospective cohort study in 2021, dividing patients into two groups based on their preference: vaginal and laparoscopic salpingectomy. The statistical analysis was conducted using STATA, employing standard statistical methods based on the distribution of variables. RESULTS: The study included 64 patients, with 34 undergoing the vaginal approach and 30 the laparoscopic route. No statistically significant differences were observed in baseline characteristics or complications between the two groups. Both surgical methods were found to be feasible. The vaginal route demonstrated a significantly shorter mean surgical time (P < 0.001). The laparoscopic route exhibited significantly less intraoperative blood loss (P < 0.001) and fewer hospitalization hours (P = 0.008). Postoperative satisfaction and pain levels did not show statistically significant differences. CONCLUSION: Vaginal salpingectomy is a feasible technique with low complication rates, making it a potential option for gynecologists to offer their patients. We propose vaginal route inclusion in the training curriculum for gynecologic surgeons during the residency program, without any subspecialty requirements.

A Four-Country Study of Strangulation-Related Alterations in Consciousness in Women Who Have Experienced Intimate Partner Violence: Co-Occurrence with Traumatic Brain Injuries and Measures of Psychological Distress.

At least one in three women experience intimate partner violence (IPV) in their lifetime. The most commonly sustained IPV-related brain injuries include strangulation-related alterations in consciousness (S-AICs) and traumatic brain injuries (TBIs). Moreover, survivors of IPV-related S-AICs and/or TBIs often demonstrate psychological distress such as depression, anxiety, and post-traumatic stress. However, the co-occurrence of S-AICs and TBIs, and whether such TBIs may be moderate to severe, has not been systematically examined, and most data have been collected from women in North America. The purpose of this study was to examine the co-occurrence of IPV-related S-AICs and TBIs across a range of geographical locations and to determine the extent to which these S-AICs are related to psychological distress. Women who had experienced physical IPV (n = 213) were included in this secondary analysis of retrospectively collected data across four countries (Canada, the United States, Spain, and Colombia). The Brain Injury Severity Assessment (BISA) was used to assess IPV-related BI across all sites. Because various questionnaires were employed to assess levels of depression, anxiety, and post-traumatic stress disorder at each site, we created a standardized composite score by converting raw scores into Z-scores for analysis. Mann-Whitney U tests and chi-square tests were conducted to examine differences between women with and without experience of S-AICs and to discover if there was a relationship between the occurrence of S-AICs and TBIs. Analysis of variance and analysis of covariance (to control for the potential confounding effects of age, education, and non IPV-related TBI) were used to compare levels of psychological distress in women who had or had not experienced S-AICs. Approximately, 67% of women sustained at least one IPV-related BI (i.e., TBI and/or S-AIC). In a subsample of women who sustained at least one IPV-related BI, approximately 37% sustained both S-AICs and TBIs, 2% sustained only S-AICs (with no TBIs), and 61% sustained TBIs exclusively (with no S-AICs). Furthermore, women who had sustained S-AICs (with or without a TBI) were more likely to have experienced a moderate-to-severe BI than those who had not sustained an S-AIC (BISA severity subscale: U = 3939, p = 0.006). In addition, women who experienced S-AICs (with or without a TBI) reported higher levels of psychological distress compared with women who never experienced S-AICs, irrespective of whether they occurred once or multiple times. These data underscore the importance of assessing for S-AIC in women who have experienced IPV and when present, to also assess for TBIs and the presence of psychological distress. Unfortunately, there were methodological differences across sites precluding cross-site comparisons. Nonetheless, data were collected across four culturally and geographically diverse countries and, therefore, highlight IPV-related BIs as a global issue that needs to be aggressively studied with policies established and then implemented to address findings.

Cutaneous glands of the striped toad, Rhinella crucifer (Wied-Neuwied, 1821) (Amphibia: Bufonidae): Histological study and bioactivities of glandular secretions.

This study investigated the morphology of Rhinella crucifer cutaneous glands, as well as the protein/peptide profiles and bioactivities of body gland secretions (BGS) and parotoid macrogland secretions (PS). The parotoid as well as dorsal and ventral skin fragments of male and female individuals were processed for histological analysis. The protein and peptide profiles of male and female gland secretions were evaluated. Male secretions were also assessed for proteolytic, trypsin inhibiting, hemagglutinating, hemolytic, antimicrobial, and anticoagulant activities. The R. crucifer skin structure presented protuberances that are clearly visible and formed by the integument, which has cutaneous glands throughout the body. An average of 438 and 333 glands were identified in males in females, respectively. No significant differences were observed in the distribution of glands across the body as well as for area and perimeter of glands. Differences were observed in protein composition between the PS and BGS from males and females, and secretions from animals collected from undisturbed and anthropogenically disturbed areas. Proteins with similarities to catalase and elongation factor 1-alpha were detected in the PS. Zymography revealed proteolytic activity in both male BGS and PS. Male BGS showed antibacterial activity against Enterococcus faecalis and Escherichia coli and anticoagulant activity, being able to prolong prothrombin time by 6.34-fold and activated partial thromboplastin time by 2.17-fold. Finally, male PS and BGS caused a maximum hemolysis degree of 1.4%. The data showed that the cutaneous secretions of R. crucifer are potentially promising for biotechnological prospecting.

Biomechanical behavior of three maxillary expanders in cleft lip and palate: a finite element study.

This study evaluated the stress distribution in the dentoalveolar and palatal bone structures during maxillary expansion in a 17-year-old male patient with bilateral cleft lip and palate (BCLP) using expanders with dental (HYRAX) and skeletal anchorage (MARPE). For the generation of the specific finite element models, cone-beam computed tomography was used, and the DICOM files were exported to Mimics 3-Matic (Materialise) and Patran (MSC Software) software. Three specific three-dimensional models were generated: A) HYRAX: conventional four-banded hyrax screw (9 mm); B) MARPE-DS: 3 miniscrews (1.8 mm diameter - 5.4 mm length) and four-banded dental anchorage; and C) MARPE-NoDS: 3 miniscrews without dental anchorage. Maxillary expansion was simulated by activating the expanders transversely 1 mm on the "X" axis. HYRAX resulted in higher levels of deformation predominantly in the dentoalveolar region. MARPE-DS showed stress in the dentoalveolar region and mainly in the center of the palatal region, at approximately 4,000 µÎµ. MARPE-NoDS exhibited evident stress only in the palatal region. High stress levels in the root anchoring teeth were observed for HYRAX and MARPE-DS. In contrast, MARPE-NoDS cause stress on the tooth structure. The stress distribution from the expanders used in the BLCP showed asymmetric expansive behavior. During the initial activation phase of expansion, the HYRAX and MARPE-DS models produced similarly high strain at the dentoalveolar structures and upper posterior teeth displacement. The MARPE-NoDS model showed restricted strain on the palate.

Green Enzymatic Synthesis of Geranyl Butyrate: Process Optimization and Mechanistic Insights.

Flavor esters are organic compounds widely used in the food industry to enhance the aroma and taste of products. However, most chemical processes for the production of these flavoring compounds use toxic organic solvents. Some organic solvents derived from petroleum can leave behind residual traces in food products, which may raise concerns about potential health risks and contamination. In this study, we employ Eversa Transform 2.0, a commercial lipase derived from the lipase from Thermomyces lanuginosus, to produce geranyl butyrate in aqueous media. The chemical process was optimized using the Taguchi method, and a conversion of 93% was obtained at the optimal reaction conditions of: 1:5 molar ratio (v/v), 15% biocatalyst load (w/w), at 50 °C, in 6 h. Classic (molecular dynamics) and quantum (density functional theory) simulations unveiled amino acid residues involved in the stabilization of the enzyme-substrate complex. Detailed QM/MM mechanistic studies identified the nucleophilic attack of the deacylation reaction as the rate-limiting step of the entire mechanism, which has a free energy barrier of 14.0 kcal/mol.

Real-world experience with pertuzumab and trastuzumab combined with chemotherapy in neoadjuvant treatment for patients with early-stage HER2-positive breast cancer: the NEOPERSUR study.

PURPOSE: HER2-targeted therapies have dramatically improved outcomes of patients with HER2-positive breast cancer (BC), as demonstrated in neoadjuvant trials. This study aims to provide real-world evidence on the use and effectiveness of combined pertuzumab, trastuzumab and chemotherapy (CT) in early-stage HER2-positive BC. METHODS: A retrospective, multicentre study was conducted on patients diagnosed with HER2-positive early BC treated with neoadjuvant pertuzumab and trastuzumab plus CT at 13 Spanish sites. The primary endpoint was pathological complete response (pCR). RESULTS: A total of 310 patients were included. Pertuzumab and trastuzumab were combined with anthracyclines and taxanes, carboplatin and docetaxel, and taxane-based CT in 77.1%, 16.5%, and 6.5% of patients, respectively. Overall, the pCR rate was 62.2%. The pCR was higher amongst patients with hormone receptor-negative tumours and with tumours expressing higher levels of Ki-67 (> 20%). After postoperative adjuvant treatment, 13.9% of patients relapsed. Those patients who did not achieve pCR, with tumours at advanced stages (III), and with node-positive disease were more likely to experience distant relapse. Median overall survival (OS) and distant disease-free survival (D-DFS) were not reached at the study end. The estimated mean OS and D-DFS times were 7.5 (95% CI 7.3-7.7) and 7.3 (95% CI 7.1-7.5) years, respectively (both were significantly longer amongst patients who achieved pCR). Grade 3-4 anti-HER2 related toxicities were reported in six (1.9%) patients. CONCLUSION: Neoadjuvant pertuzumab and trastuzumab plus CT achieve high pCR rates in real-life patients with HER2-positive early BC, showing an acceptable safety profile. Innovative adjuvant strategies are essential in patients at high risk of distant disease recurrence.

Trends in Mercury Contamination Distribution among Human and Animal Populations in the Amazon Region.

Mercury contamination in the Amazon arising from both natural sources and intensive mining activities in the region is a significant public health concern. This metal is used to separate Au from sediments. Accordingly, this study aimed to assess the impact of mining on mercury contamination in the animal and human populations of the Amazon. This overall objective was pursued through a systematic review of the existing literature to assess the impact of Hg and identify gaps in geographic coverage arising from this assessment. Herein, we employed PECO and PRISMA-ScR protocols to select articles published between 2017 and 2023 based on projected points on a map within the biogeographic boundaries of the Amazon. We found that mercury concentrations increase with trophic levels, reaching high values of 3.7 µg/g in the muscles of predatory fish and 34.9 µg/g in human hair. The mean level of mercury in human hair in the whole (Amazon) region exceeds 6 µg/g, surpassing tolerance levels. Although mining regions show high concentrations of Hg, the highest incidence was observed among populations with fish-based diets. It was concluded that continuous research and monitoring of fish in the region are required in order to accurately assess the risk associated with Hg contamination, especially since fish are the main source of protein in this region.

Effects of Teosinte Flour (Dioon mejiae) on Selected Physicochemical Characteristics and Consumer Perceptions of Gluten-Free Cocoa Cookies Formulated with Mung Bean (Vigna radiata) Flour.

Dioon mejiae, or teosinte, is a living fossil tree discovered in Olancho, Honduras, whose seeds have a desirable nutritional profile that can provide health benefits. As a result, the objective of this study was to evaluate the effects of teosinte flour obtained from seeds on selected physicochemical characteristics and consumer perceptions of gluten-free cocoa cookies formulated with mung bean (Vigna radiata) flour. Gluten-free cocoa cookies were prepared with different levels of teosinte flour (0%, 70%, 80%, 90%, and 100% by weight of mung bean flour) in substitution of mung bean flour. The cookies were evaluated for texture hardness, color (L*, a*, b*), moisture content, and water activity. Sensory acceptability of appearance, color, texture, aroma, flavor, and overall quality of cocoa cookies was rated by 175 consumers using a "yes/no" binomial scale. Overall liking was evaluated using a 9-point hedonic scale. Purchase intent was evaluated with a "yes/no" binomial scale. The levels of teosinte flour did not significantly affect the acceptability of appearance, color, texture, flavor, aroma, and overall quality, and neither the overall liking nor the purchase intent responses. However, the texture attribute had the lowest % acceptability response among all sensory attributes. The addition of teosinte flour did not affect water activity and color (L*, a*, b*), whereas it decreased the texture hardness (g force), producing softer cookies. Cocoa cookies stayed acceptable even at 100% teosinte flour addition (70% acceptability; mean overall liking = 5.69). Teosinte flour has an excellent nutritional profile that could be practically applied in baked goods.

The impact of methylene blue photosensitizer, aPDT and a calcium hydroxide-based paste on the physicochemical and mechanical characteristics of root canal dentin and the bonding interface of fiberglass posts.

PURPOSE: To investigate the influence of methylene blue (MB)-mediated antimicrobial photodynamic therapy (aPDT) and calcium hydroxide (CH) medication on the mechanical characteristics, degree of conversion (DC), quantification, and volume of gaps at the adhesive interface of glass fiber posts (GFPs) luted to distinct thirds of root canal dentin. Additionally, the microhardness (MH), elastic modulus (Eit), morphology, and chemical structure of the intraradicular dentin were assessed. MATERIALS AND METHODS: 6 experimental groups were formed by sorting 102 bovine incisors. Canals receiving deionized water irrigation as a negative control; canals receiving deionized water irrigation and filled with CH as a positive control; groups treated with CH + MB at 50 and 100 mg/L without irradiation; and groups treated with CH + MB at 50 and 100 mg/L irradiated by red laser for 60 s (660 nm; 100 mW; 6.5 J; 72 J/cm2). MH, Eit, and DC properties were evaluated for both the resin cement layer and root dentin substrate (n = 8). Volume and quantification of gaps at the bonding interface (n = 6), and dentin morphology and chemical content were investigated (n = 3). Data were analyzed using a repeated-measures 2-way ANOVA followed by Tukey post hoc analysis (α = 0.05). RESULTS: The distinct intraradicular thirds and treatment with MB-mediated aPDT, whether activated or not, in combination with CH, had a significant impact on the mechanical characteristics of the root dentin. This effect was also observed in the MH, Eit, DC, quantification, and volume of gaps at the luting interface (P < .05). In general, a higher concentration of MB, whether activated by a red laser or not, led to lower values in the mechanical properties of the root dentin, as well as in MH, Eit, and DC at the adhesive interface (P < .05). Additionally, these groups exhibited higher values for quantification and volume of gaps at the luting substrate (P < .05). Scanning electron micrographs and energy dispersive X-ray spectra showed qualitative similarity among all groups, except for the negative experimental control group. CONCLUSIONS: MB-mediated aPDT at 50 mg/L, in combination with CH, demonstrated favorable physico-chemical and mechanical characteristics in intraradicular dentin, along with satisfactory mechanical features and the adhesive interface integrity for GFPs at all intraradicular depths. CLINICAL SIGNIFICANCE: MB-mediated aPDT at a concentration of 50 mg/L combined to CH medication represents a suitable choice for photosensitization in the context of intracanal disinfection following the biomechanical procedure and prior to luting of intraradicular restorations.

TLC and HPLC methods for the determination of plumbagin for the diagnosis of poisoning by Plumbago scandens L.

Plumbago scandens L. (Plumbaginaceae) occurs in all regions of Brazil. It has been described as toxic to cattle and goats. Caustic lesions in the upper digestive tract characterize poisoning. P. scandens contains a naphthoquinone named plumbagin, which presents high cytotoxic activity. Plumbago auriculata Lam., a widely used ornamental plant, is considered potentially toxic, but there is limited data about its toxicity. This work aimed to validate analytical methodologies for determining the levels of plumbagin in samples of leaves, stems, and rumen content to be used as an auxiliary chemical marker in the laboratory diagnosis of intoxication. One methodology used thin layer chromatography (TLC), and another used high-performance liquid chromatography (HPLC). The presence of palisade grass (Urochloa brizantha (Hochst. ex A.Rich.) R.D.Webster), Guinea grass (Megathyrsus maximus (Jacq.) B.K.Simon & S.W.L.Jacobs), corn silage, and rumen content did not interfere with plumbagin in the two methodologies. The TLC methodology generates qualitative results but is simple to implement and has a low cost. The HPLC methodology showed a limit of detection (LOD) of 0.01 µg/mL and a limit of quantification (LOQ) of 0.05 µg/mL. Leaf and stem samples of P. scandens evaluated showed high levels of plumbagin (0.261 ± 0.087 % and 0.327 ± 0.055 %, respectively). In contrast, leaves of P. auriculata did not show detectable levels of the toxin, and some stem samples showed low levels (up to 0.000114 %). Thus, these methodologies can be used to confirm or rule out the consumption of P. scandens in rumen content from animals suspected of poisoning.

Antimicrobial Resistance and Antibiotic Consumption in a Secondary Care Hospital in Mexico.

BACKGROUND: Antimicrobial resistance is a global health problem, due to morbidity, mortality, and healthcare costs. The misuse of antimicrobials is the main cause of antimicrobial resistance. The aim of this study was to report antimicrobial resistance and antibiotic consumption in a secondary care hospital in Mexico. METHODS: Within a cross-sectional study, antimicrobial resistance data on ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) and antibiotic consumption from 2020 to 2022 were collected. Antimicrobial resistance was reported based on percentages of resistance and consumption was analyzed using the defined daily dose (DDD)/100 bed days and the AWaRe (Access, Surveillance, Reservation) antibiotic group. RESULTS: Antibiotic consumption in 2020, 2021 and 2022 was 330, 175 and 175 DDD/100 beds day, respectively. The rate of ceftriaxone resistance in E. coli (n = 526) and K. pneumoniae (n = 80) was 76% and 69%, respectively, the rate of carbapenem resistance in A. baumannii (n = 168) and P. aeruginosa (n = 108) was 92% and 52%, respectively; the rate of oxacillin resistance in S. aureus (n = 208) was 27%; and the rate of vancomycin resistance in E. faecium (n = 68) was 47%. CONCLUSION: The reported results are congruent with global estimates of antibiotic resistance and consumption, providing an overview that could generate actions for antimicrobial optimization at the local and regional levels.

Cross talk between the liver microbiome and epigenome in patients with metabolic dysfunction-associated steatotic liver disease.

BACKGROUND: The pathogenesis of MASLD (metabolic dysfunction-associated steatotic liver disease), including its severe clinical forms, involves complex processes at all levels of biological organization. This study examined the potential link between the liver microbiome profile and epigenetic factors. METHODS: Liver microbial DNA composition was analysed using high throughput 16S rRNA gene sequencing in 116 individuals, with 55% being female, across the spectrum of liver disease severity. Total activity of histone deacetylases (HDACs) and acetyltransferases (HATs) was assayed in nuclear extracts from fresh liver samples. In addition, we measured the global 5-hydroxymethylcytosine (5-hmC) levels of liver DNA. FINDINGS: Patients with MASLD showed a 2.07-fold increase (p = 0.013) in liver total HAT activity. Moreover, a correlation was observed between liver total HAT activity and the score for histological steatosis (Spearman's R = 0.60, p = 1.0E-3) and disease severity (R = 0.40, p = 2.0E-2). Liver HAT and HDAC activities also showed associations with the abundance of several liver bacterial DNAs. Additionally, liver global levels of 5-hmC showed negative correlation with the read number of Bacteroidetes (R = -0.62, p = 9.3E-4) and Gammaproteobacteria (R = -0.43, p = 3.2E-2), while it was positively correlated with the abundance of Acidobacteria (R = 0.42, p = 4.1E-2) and Actinobacteria (R = 0.47, p = 1.8E-2). INTERPRETATION: The host liver epigenome, including the activity of enzymes involved in maintaining the balance between protein acetylation and deacetylation and the global DNA hydroxy-methylation status, may be the target of microbial signals. FUNDING: Agencia Nacional de Promoción Científica y Tecnológica, FonCyT.

Sterilization of Human Amniotic Membrane Using an Ozone Hydrodynamic System.

Human amniotic membrane (hAM) is an important biomaterial for Tissue Engineering, due to its great regenerative properties and potential use as a scaffold. The most used procedure to sterilize biomaterials is gamma-irradiation, but this method can affect several properties, causing damage to the structure and reducing the growth factors. The present work evaluated the efficiency of a new method based on ozonated dynamic water for hAM sterilization. HAM fragments were experimentally contaminated with Staphylococcus aureus, Escherichia coli, Candida albicans, Staphylococcus epidermidis, and Clostridium sporogenes (106 CFU/mL) and submitted to sterilization process for 5, 10 and 15 min. The analyses did not reveal microbial activity after 10 min for S. aureus and C. sporogenes and after 15 min for E. coli and S. epidermidis. The microbial activity of C. albicans was reduced with the exposure time increase, but the evaluated time was insufficient for complete sterilization. The depyrogenation process was investigated for different ozonation times (15, 20, 25, 30, and 35 min) to evaluate the ozone sterilization potential and presented promising results after 35 min. The ozone effect on hAM structure was evaluated by histological analysis. A decrease in epithelium average thickness was observed with the exposure time increase. Furthermore, some damage in the epithelium was observed when hAM was exposed for 10 and 15 min. It can indicate that ozone, besides being effective in sterilization, could promote the hAM sample's de-epithelization, becoming a possible new method for removing the epithelial layer to use hAM as a scaffold.