Exploring the potential of phenolic and antioxidant compounds in new Rosaceae fruits.

BACKGROUND: Rosaceae fruits have been used in traditional medicine for the prevention and treatment of diseases. However, these fruits have not extensively been studied regarding their phenolic composition. Thus, this research focuses on the determination of phenolic compounds by high-performance liquid chromatography electrospray ionization time-of-flight mass spectrometry, flavan-3-ols by high-performance liquid chromatography with fluorescence detection, and the antioxidant activity by 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power of the fruits of five species of genera Crataegus and Sorbus (Rosaceae). RESULTS: We found a total of 71 phenolic compounds from which 30 were identified in these berries for the first time. Crataegus monogyna and Crataegus laciniata revealed higher total phenolic and flavan-3-ol contents than the other species and the highest antioxidant activities. CONCLUSIONS: Therefore, the fruits evaluated have demonstrated to be important sources of bioactive compounds with huge potential for being used in nutraceutical or food scopes. Additional studies could be needed to evaluate the influence of the different production areas on the phenolic content. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Xerophytic Lichens from Gypsiferous Outcrops of Arid Areas of Andalusia as a Source of Anti-Phytopathogenic Depsides.

In a survey to evaluate the potential of lichens associated with gypsum areas as sources of new antifungal metabolites, six species of lichens were collected in the gypsum outcrops of the Sorbas Desert (Diploschistes ocellatus and Seirophora lacunosa) and the Tabernas Desert (Cladonia foliacea, Acarospora placodiformis, Squamarina lentigera and Xanthoparmelia pokornyi) in southern Spain. Raw lichen acetone extracts were tested against a panel of seven phytopathogenic fungi, including Botrytis cinerea, Colletotrichum acutatum, Fusarium oxysporum f.sp cubense TR4, Fusarium ploriferaum, Magnaporthe grisea, Verticillium dahliae and Zymoseptoria tritici. Active extracts of Cladonia foliacea, Xanthoparmelia pokornyi and Squamarina lentigera were analyzed by HPLC-MS/MS and Molecular Networking to identify possible metabolites responsible for the antifungal activity. A total of ten depside-like metabolites were identified by MS/MS dereplication and NMR experiments, of which one was a new derivative of fumaroprotocetraric acid. The compounds without previously described biological activity were purified and tested against the panel of fungal phytopathogens. Herein, the antifungal activity against fungal phytopathogens of 4'-O-methylpaludosic acid, divaricatic acid and stenosporic acid is reported for the first time. Stenosporic and divaricatic acids displayed a broad antifungal spectrum against seven relevant fungal phytopathogens in a micromolar range, including the extremely resistant fungus F. oxysporum f. sp. cubense Tropical Race 4 (TR4). 4'-O-methylpaludosic acid exhibited specific antifungal activity against the wheat pathogen Z. tritici, with an IC50 of 38.87 µg/mL (87.1 µM) in the absorbance-based assay and 24.88 µg/mL (55.52 µM) in the fluorescence-based assay.

Evaluation of Anticancer Activity of 76 Plant Species Collected in Andalusia (Spain) against Lung Cancer Cells.

Every year, cancer kills millions of people around the world. Finding more selective anticancer agents is essential to improve the low survival rates of patients with metastatic cancers. Since the research of natural products is a valuable approach to the discovery of new compounds and the Iberian flora offers a rich source of unstudied plants, we have carried out a random screening of 76 plant species from 43 families collected in Andalusia (South of Spain). Using non-malignant cells (HaCaT) and lung cancer cells (A549), we found that the extract from Arum italicum Mill. subsp. italicum (Araceae), Mandragora autumnalis Bertol. (Solanaceae), Rhamnus alaternus L. (Rhamnaceae), and Lomelosia simplex (Desf.) Raf. subsp. dentata (Jord. & Fourr.) Greuter & Burdet (Dipsacaceae) showed selective cytotoxicity against lung cancer cells. Extracts of plant species belonging to the Iridaceae family showed high selective activity against cancer cells, highlighting that the Xiphion xiphium (L.) M.B. Crespo, Mart.-Azorín & Mavrodiev flower extract was more selective against lung cancer cells than the standard anticancer drugs, cisplatin and 5-fluorouracil. This extract also showed modest selective cytotoxicity against bladder carcinoma cells (T24). The number of cells in the G1 phase increased after treatment with the extract from Xiphion xiphium. Our research indicates that various plants are potential sources for the isolation and development of new anticancer drugs.

Wild Edible Plants of Andalusia: Traditional Uses and Potential of Eating Wild in a Highly Diverse Region.

A review of ethnobotanical sources focused on traditionally-used wild food plants in Andalusia (southern Spain), one of the most biodiverse regions in Europe, is carried out. With 21 original sources plus some previously unpublished data, the dataset shows a high diversity of these traditional resources, reaching 336 species or c. 7% of the total wild flora. Cultural aspects related to the use of some species are discussed and data are compared with similar works. The results are discussed through the lens of conservation and bromatology. For 24% of the edible plants, informants also mentioned a medicinal use (achieved by consuming the same part of the plant). In addition, a list of 166 potentially edible species is provided based on a review of data from other Spanish territories.

The rise and fall of mandrake in medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Mandrake (Mandragora sp.) is one of the most famous medicinal plants. It has been in continuous medical use throughout written history and is still in use today in popular medicine. AIM OF THE STUDY: Mandrake derived drugs once played an important role in medicine and in magical practices. Today, the role of mandrake in popular medicine is marginal. However, natural products present in mandrake such as atropine and scopolamine, as well as their semi synthetic derivatives continue to hold and important role in medicine. Here we aim to trace the development of historical rationales and scientific events that led to the abandonment of mandrake as a medicine. MATERIALS AND METHODS: We review the medicinal uses of mandrake drugs since antiquity in an attempt to pinpoint use patterns that were popular in certain periods of time and others that are more general. We compare the uses from the native territories to those from regions where the plant got introduced and use literature reporting mandrake's chemistry and pharmacology in order to explain the diachronic changes of use patterns. RESULTS AND CONCLUSION: We found information about 88 different medicinal uses for mandrake, grouped into 39 conditions. According to the number of different medicinal uses, the most versatile period was the medieval (37), followed by the Renaissance (31), the classical (27), and the modern period (21). Considering the higher number of textual sources and use-records collected for the Renaissance period, the decrease of versatility in comparison to the medieval period appears robust. This seems to indicate a more consolidated use pattern, that might be conditioned by the reproduction of classic textual sources as well as by a less experimental approach and reduced popularity of mandrake in medicine. The introduction of the volatile anaesthetics with more reliable narcotic effects set the seal on using mandrake in surgery but opened the way for atropine being used as a prophylactic and antidote during surgical interventions.

Structural and evolutionary analysis unveil functional adaptations in the promiscuous behavior of serum albumins.

Promiscuous activities have been related to the capacity to catalyze reactions different from those a protein has evolved to sustain. In this work, we rethought the serum albumin's promiscuous behavior using evolutionary and structural analysis. We found that the cross aldol condensation of acetone and p-formylbenzonitrile is a promiscuous reaction conserved in humans serum albumin and in closely related albumins from other mammals. Evolutionary analysis indicates that the residues involved in this promiscuous reaction are evolving under positive selection, an evolutionary pattern indicating a putative functional adaptation. Also, key residues are located in an evolutionary conserved cavity connected with the protein surface with an also conserved tunnel and mutations involving these residues are described in human diseases. Overall, our results suggest that albumin could have evolved to sustain a still unknown biological function among the many others it maintains. Our results could contribute to better characterize the serum albumin family and raise questions about the evolution of protein promiscuity and function.

In search of traces of the mandrake myth: the historical, and ethnobotanical roots of its vernacular names.

BACKGROUND: Mandrake (Mandragora spp.) is one of the most famous medicinal plant in western cultures since Biblical times and throughout written history. In many cultures, mandrake is related to magic and witchcraft, which is said to have a psychosomatic effect (especially when mandrake contains narcotic compounds) in addition to the pharmacological influence, as occurs with other narcotic magical plants. Due to its unique properties and related myths, it is not surprising that this plant has many names in many languages. METHODS: This paper presents an attempt to reconstruct the historical, ethnobotanical, and folkloristic roots of 292 vernacular names of Mandragora spp. in forty-one languages. We used the plant's morphological data, philology, myths and legends, medicinal properties and uses, as well as historical evidence and folkloric data, to explain meaning, origin, migration, and history of the plant's names. RESULTS: The names were classified into the following main categories: Derivatives of mandragora (19 languages), alraun (7) and of yabroukh (5). The salient groups of the plant's vernacular names are related to: Anthropomorphism (33 names in 13 languages); Similarity to other plants (28/9); Supernatural agents (28/9); Narcotic effects (21/8); Leaves, fruits, and seeds (21/8); Aphrodisiac properties (17/10); Use of a dog (15/9); Gallows (14/5); Black magic, sorcery, witchcraft (13/8), and Medicinal use (11/7). CONCLUSIONS: This frequency distribution of the mandrake's vernacular names reflects its widespread reputation as related to the doctrine of signatures, beliefs in its supernatural, natural, and mythic powers, and to a lesser extent, its uses in magic and medicine. A spatiotemporal analysis of the mandrake's names supports the old idea that the pulling ceremonies for this plant originated in the Near East and that various other myths related to this plant may have originated in different places and periods.

The Doctrine of Signatures in Israel-Revision and Spatiotemporal Patterns.

The present survey includes forty-three plant species with present-day medicinal applications that can be related to the Doctrine of Signatures (DoS). The main uses are for jaundice (33.3%), kidney stones (20%), and as an aphrodisiac (8%). Ten Doctrine of Signature uses (22.2%) are endemic (to Israel and Jordan); while none of these plant species are endemic to the region at all, their DoS uses are endemic. Summing up of all these data reveals that 73.2% of all uses found in present-day Israel could be considered as related to Muslim traditional medicine. About one quarter (24.4%) of the DoS uses are also common to Europe, and some (8.8%) to India. The two adventive species with DoS uses serve as evidence that the DoS practice is not necessarily based solely on its historical background but is still evolving locally in accordance with changes in the local flora. The current broad geographic distribution of many of the doctrine's uses may serve as indirect evidence of its current prevalence, and not just as a vestigial presentation of ancient beliefs.

Setup of an Ultrasonic-Assisted Extraction to Obtain High Phenolic Recovery in Crataegus monogyna Leaves.

Hawthorn leaves are a rich source of phenolic compounds that possess beneficial activities for human health. Ultrasonic-assisted extraction (UAE) is an extraction technique frequently used for the isolation of phenolic compounds in plants. Thus, in this study, a Box-Behnken design was used to optimize UAE conditions such as the percentage of acetone, the extraction time and solvent-to-solid ratio (v/w) in order to obtain the maximum content of total compounds by Folin-Ciocalteu and the maximum in vitro antioxidant activity by DPPH, ABTS and FRAP assays in Crataegus monogyna leaves. The optimum conditions to obtain the highest total phenolic content and antioxidant activities were 50% acetone, 55 min and 1/1000 (w/v). A total of 30 phenolic compounds were identified and quantified in C. monogyna leaf extract obtained at these optimum UAE conditions. HPLC-MS allows the identification and quantification of 19 phenolic compounds and NP-HPLC-FLD analyses showed the presence of 11 proanthocyanidins. According to the results, the most concentrated phenolic compounds in C. monogyna leaf extract obtained at optimum UAE conditions were phenolic acid derivatives such as protocatechuic acid-glucoside, dihydroxy benzoic acid pentoside and chlorogenic acid, flavones such as 2″-O-rhamnosyl-C-hexosyl-apigenin, flavonols such as hyperoside and isoquercetin and proanthocyanidins such as monomer and dimer. As a result, the optimized UAE conditions could be used to obtain an extract of C. monogyna leaves enriched with phenolic compounds.

Ethnozoology among the Berbers: pre-Islamic practices survive in the Rif (northwestern Africa).

BACKGROUND: Ethnozoological knowledge is less documented than ethnobotanical. With this field study, we aim to record and analyze the Riffian Berber knowledge about the use of animals in traditional human and veterinary medicine. Our research question is what is their knowledge of ethnozoological practices? METHODS: We performed semi-structured interviews with local inhabitants in Riffian vernacular language. The reliability of the sampling effort was assessed by a rarefaction curve. Data were compared with previous studies in order to determine the geographical and historical extensions of described uses and possible conservation implications for the species used. RESULTS: We obtained information regarding 107 ethnozoological uses based on 197 use reports. Among the 31 species used, mammals were most frequently cited. Diseases related to the traditional medicinal system were most frequently treated with these resources, as well as those of the respiratory, digestive, and musculoskeletal systems. Thirty percent of uses are associated with magico-religious practices. Only three of the species used are threatened at the global level, two of them extinct in the study area, indicating low potential damage to regional biodiversity from current practices utilizing native animals. Within modern Morocco, Riffians have continued practicing ethnozoological uses anathema to Islam, like the consumption of animals considered impure (dogs, jackals, wild boars, and hyenas). CONCLUSIONS: The use of primarily mammalian species and of many animal body parts is likely related to the Berber belief in homology between the area of the human body in which the ailment occurs and the corresponding animal body part. These findings unveil the nature of ethnozoological practices, highly linked to folklore and culture-bound conditions, and lacking in the Western empirical rationale for nearly one third of reported uses. The consumption of animals considered impure according to Islam was probably initiated before the conquering of the Maghreb by Arabs in the seventh century and was maintained through the secular isolation of Riffians in mountain areas. This can reflect traditional healing habits being maintained over thousands of years.

Multifunctional Titanium Surfaces for Orthopedic Implants: Antimicrobial Activity and Enhanced Osseointegration.

The use of implants in orthopedics and dental practice is a widespread surgical procedure to treat diverse diseases. However, peri-implantitis due to infections and/or poor osseointegration can lead to metallic implant failure. The aim of this study was to develop a multifunctional coating on titanium (Ti) surfaces, to simultaneously deal with both issues, by combining antibacterial silver nanoparticles (AgNPs) and regenerative properties of lactoferrin (Lf). A simple and cost-effective methodology that allows the direct multifunctionalization of Ti surfaces was developed. The modified surfaces were characterized by atomic force microscopy (AFM), X-ray photoelectron spectroscopy, and contact angle measurements. Additionally, in vitro preosteoblast cell adhesion, cell viability, and differentiation were evaluated. The antibacterial capability of the surfaces was tested against Staphylococcus aureus as a prosthesis infection model strain. Our results showed that Lf adsorbed on both Ti surfaces and Ti surfaces with adsorbed AgNPs. Simultaneously, the presence of Lf and AgNPs notably improved preosteoblast adhesion, proliferation, and differentiation, whereas it reduced the bacterial colonization by 97.7%. Our findings indicate that this simple method may have potential applications in medical devices to both improve osseointegration and reduce bacterial infection risk, enhancing successful implantation and patients' quality of life.

Revenant: a database of resurrected proteins.

Revenant is a database of resurrected proteins coming from extinct organisms. Currently, it contains a manually curated collection of 84 resurrected proteins derived from bibliographic data. Each protein is extensively annotated, including structural, biochemical and biophysical information. Revenant contains a browse capability designed as a timeline from where the different proteins can be accessed. The oldest Revenant entries are between 4200 and 3500 million years ago, while the younger entries are between 8.8 and 6.3 million years ago. These proteins have been resurrected using computational tools called ancestral sequence reconstruction techniques combined with wet-laboratory synthesis and expression. Resurrected proteins are commonly used, with a noticeable increase during the past years, to explore and test different evolutionary hypotheses such as protein stability, to explore the origin of new functions, to get biochemical insights into past metabolisms and to explore specificity and promiscuous behaviour of ancient proteins.

Ethnobotanical study of medicinal plants in the Adrar Province, Mauritania.

ETHNOPHARMACOLOGICAL RELEVANCE: Mauritania is a country in which few ethnobotanical studies have been conducted and consequently the ethnomedical data is scarce. Since the geographical region reflects the transition between tropical and Northern Africa, influenced by the Mediterranean floristic region, the traditional knowledge was influenced by several cultures from tropical Africa as well as Arab, Berber and Islamic societies. AIM OF THE STUDY: This paper aims to explore and compile the diversity of ethnomedical knowledge in one of the regions of Mauritania and to compare the data with similar studies from surrounding territories. MATERIALS AND METHODS: Surveys and interviews were carried out in 11 villages of Adrar province in northern Mauritania. Data were collected through open, semi-structured interviews (with individuals and focus groups). A sample of 120 people aged between 20 and 70 years, including 24 herbalists and 28 traditional healers was included. RESULTS: Ethnomedical data for 68 plant species belonging to 27 families were obtained. They are used to treat 50 health conditions grouped in 14 pathological groups. Remedies for digestive system disorders, skin problems and respiratory ailments were among the most frequent indications. Leaves were the most frequently used plant part and remedies generally used as a powder for the various applications and a total of 2'317 use reports were gathered. About 55% of the reported species were not cited previously in the literature focusing on Mauritania and neighbouring countries. Moreover, only 6 species are also cited by Ibn al-Baytar (13th century CE). CONCLUSIONS: This work shows a promising perspective for future studies, shedding light on the richness and the risk for conservation of traditional knowledge of herbal medicine in Mauritania.

DisProt: intrinsic protein disorder annotation in 2020.

The Database of Protein Disorder (DisProt, URL: https://disprot.org) provides manually curated annotations of intrinsically disordered proteins from the literature. Here we report recent developments with DisProt (version 8), including the doubling of protein entries, a new disorder ontology, improvements of the annotation format and a completely new website. The website includes a redesigned graphical interface, a better search engine, a clearer API for programmatic access and a new annotation interface that integrates text mining technologies. The new entry format provides a greater flexibility, simplifies maintenance and allows the capture of more information from the literature. The new disorder ontology has been formalized and made interoperable by adopting the OWL format, as well as its structure and term definitions have been improved. The new annotation interface has made the curation process faster and more effective. We recently showed that new DisProt annotations can be effectively used to train and validate disorder predictors. We believe the growth of DisProt will accelerate, contributing to the improvement of function and disorder predictors and therefore to illuminate the 'dark' proteome.

The historical development of pharmacopoeias and the inclusion of exotic herbal drugs with a focus on Europe and Brazil.

ETHNOPHARMACOLOGICAL RELEVANCE: In spite of the rich bio-cultural diversity found in the Neotropics relatively few herbal drugs native to South America are included in the global pharmacopoeia. MATERIAL AND METHODS: In the attempt to historically explain the inclusion of herbal drugs into official pharmacopoeias we consider the disparate epidemiology and cultural evolution of the New and the Old World. We then trace the development of pharmacopoeias and review forces that worked towards and against the synchronization of pharmacopoeias and highlight the role of early chemical and pharmacological studies in Europe. Finally, we compare the share of exotic and native herbal drug species included in the Brazilian Pharmacopoeia with the share of exotic and native species included in the European Pharmacopoeia as well as those used for products registered with ANVISA. RESULTS: The domination of Eurasian herbal drugs in the European Pharmacopoeia seems to be conditioned by the geographical extension of Eurasia, which facilitated the interchange of materia medica and the creation of a consensus of use since ancient times. At the time of the Conquest the epidemiology of the Amerindian populations resembled that of pre-agriculturalist societies while no written consensus around efficacious medicine existed. Subsequently, introduced and well-tried plant species of the Old World gained therapeutic importance in the New World. CONCLUSION: The research focus in Europe and the US resulted in a persistence of herbal drugs with a historic importance in the European and US pharmacopoeias, which gained a status as safe and efficacious. During the last decades only few ethnopharmacological field-studies have been conducted with indigenous Amerindian groups living in the Brazilian Amazon, which might be attributable to difficulties in obtaining research permissions. Newly adopted regulations regarding access to biodiversity and traditional knowledge as well as the simplified procedure for licencing herbal medicinal products in Brazil prospects an interesting future for those aiming at developing herbal medicine based on bio-cultural diversity and respecting the protocols regulating benefit sharing.

Bioinformatics calls the school: Use of smartphones to introduce Python for bioinformatics in high schools.

The dynamic nature of technological developments invites us to rethink the learning spaces. In this context, science education can be enriched by the contribution of new computational resources, making the educational process more up-to-date, challenging, and attractive. Bioinformatics is a key interdisciplinary field, contributing to the understanding of biological processes that is often underrated in secondary schools. As a useful resource in learning activities, bioinformatics could help in engaging students to integrate multiple fields of knowledge (logical-mathematical, biological, computational, etc.) and generate an enriched and long-lasting learning environment. Here, we report our recent project in which high school students learned basic concepts of programming applied to solving biological problems. The students were taught the Python syntax, and they coded simple tools to answer biological questions using resources at hand. Notably, these were built mostly on the students' own smartphones, which proved to be capable, readily available, and relevant complementary tools for teaching. This project resulted in an empowering and inclusive experience that challenged differences in social background and technological accessibility.

The genus Datura L. (Solanaceae) in Mexico and Spain - Ethnobotanical perspective at the interface of medical and illicit uses.

ETHNOPHARMACOLOGICAL RELEVANCE: The different species of the genus Datura have been used traditionally by some pre-Columbian civilizations, as well as in medieval rituals linked to magic and witchcraft in both Mexico and Europe. It is also noteworthy the use of different alkaloids obtained from the plants for medicinal purposes in the treatment of various groups of diseases, especially of the respiratory and muscularskeletal systems. AIM OF THE STUDY: A review of the ethnobotanical uses of the genus Datura in Mexico and Spain has been conducted. We focus on the medicinal and ritualistic uses included in modern ethnobotanical studies, emphasizing the historical knowledge from post-colonial American Codices and medieval European texts. Datura's current social emergency as a drug of recreation and leisure, as well as its link to crimes of sexual abuse is also considered. The work is completed with some notes about the distribution and ecology of the different species and a phytochemical and pharmacological review of Datura alkaloids, necessary to understand their arrival in Europe and the ethnobotanical uses made since then MATERIALS AND METHODS: A literature review and compilation of information on traditional medicinal uses of the genus has been carried out from the main electronic databases. Traditional volumes (codices) have also been consulted in libraries of different institutions. Consultations have been made with the National Toxicological Services of Spain and Mexico for toxicological data. RESULTS: A total of 118 traditional uses were collected in both territories, 111 medicinal ones to be applied in 76 conditions or symptoms included in 13 pathological groups. Although there are particular medicinal uses in the two countries, we found up to 15 similar uses, of which 80% were previously mentioned in post-Colonial American codices. Applications in the treatment of asthma and rheumatism are also highlighted. Apart from medicinal uses, it is worth noting their cultural and social uses, in the case of Mexico relating to diseases such as being scared, astonishment or falling in love, and in the case of Spain, as a recreational drug and lately, for criminal purposes. CONCLUSIONS: This review highlights the variety of uses traditionally given to the different species in both territories. The fact that most of the coincident or similar uses in both countries also appear in the classical codices can be found an example of the flow, not only of the plants from America to Europe, but also of their associated information. It is also relevant that particular uses have derived in both countries, reflecting the difference in the cultural factors and traditions linked to rituals and cultural practices. Finally, the significant growth of Datura consumption in recent years as a drug of leisure and recreation, as well as in crimes of sexual submission, should be considered as research of maximum relevance in the field of forensic botany and toxicology.

Self-sterilizing ormosils surfaces based on photo-synzthesized silver nanoparticles.

Medical device-related infections represent a major healthcare complication, resulting in potential risks for the patient. Antimicrobial materials comprise an attractive strategy against bacterial colonization and biofilm proliferation. However, in most cases these materials are only bacteriostatic or bactericidal, and consequently they must be used in combination with other antimicrobials in order to reach the eradication condition (no viable microorganisms). In this study, a straightforward and robust antibacterial coating based on Phosphotungstate Ormosil doped with core-shell (SiO2@TiO2) was developed using sol-gel process, chemical tempering, and Ag nanoparticle photoassisted synthesis (POrs-CS-Ag). The coating was characterized by X-ray Fluorescence Spectroscopy (XRF), Field Emission Scanning Electron Microscopy (FE-SEM), Atomic Force Microscopy (AFM) and X-ray Photoelectron Microscopy (XPS). The silver free coating displays low antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa, in opposition to the silver loaded ones, which are able to completely eradicate these strains. Moreover, the antimicrobial activity of these substrates remains high until three reutilization cycles, which make them a promising strategy to develop self-sterilizing materials, such as POrs-CS-Ag-impregnated fabric, POrs-CS-Ag coated indwelling metals and polymers, among other materials.

Two-step biocompatible surface functionalization for two-pathway antimicrobial action against Gram-positive bacteria.

The use of indwelling devices has emerged as a frequent and often life-saving medical procedure. However, infection in prosthetic surgery is one of the most important and devastating complications. Once the biofilm has been formed, its eradication is extremely difficult, due to an increased resistance to host defense and conventional antimicrobials. Thus, the design of novel strategies for inhibiting the bacterial adhesion on implantable devices is a key point for successful surgical procedures. In this work, the development of a simple two-step protocol to prepare surfaces able to prevent the bacterial growth was successfully achieved. The surface-modification design includes a combined approach involving the multi-functionalization of Ti surfaces with silver nanoparticles (AgNPs) and/or ampicillin (AMP). The surface chemistry involved in AMP adsorption on titanium and silver surfaces was elucidated for the first time, thus establishing the basis for the further anchoring of other antibacterial compounds having similar functional groups. Our results show that the antibiotic binds to the titanium surface through covalent interactions between the COOH groups in AMP and the OH groups of the native TiO2 on the surface, although electrostatic interactions between protonated AMP and negatively charged TiO2 can also contribute to the antibiotic anchoring to the surface. The AMP immobilization on the AgNPs is carried out by thiolate-like bonds. The ß-lactam ring functionality is preserved after the adsorption process, since the Ti-AgNPs-AMP surface was able to decrease the bacterial viability in more than 80%. Moreover, the antimicrobial capacity is maintained over time due to a two-pathway antibacterial mechanism: death by contact (AMP) and death by release (AgNPs). The effect of AMP prevails on AgNPs at early stages of bacterial adhesion, while AgNPs are responsible for sustaining the relatively low but steady release of Ag(I), preserving the bacteriostatic activity of the surface over time. This effect would contribute to prevent infections due to sessile cells on indwelling devices, powering the action of the immune system and the conventional antibiotics usually dosed in implanted patients.