RESUMO
Osteosarcoma (OS) cancer treatments include systemic chemotherapy and surgical resection. In the last years, novel treatment approaches have been proposed, which employ a drug-delivery system to prevent offside effects and improves treatment efficacy. Locally delivering anticancer compounds improves on high local concentrations with more efficient tumour-killing effect, reduced drugs resistance and confined systemic effects. Here, the synthesis of injectable strontium-doped calcium phosphate (SrCPC) scaffold was proposed as drug delivery system to combine bone tissue regeneration and anticancer treatment by controlled release of methotrexate (MTX) and doxorubicin (DOX), coded as SrCPC-MTX and SrCPC-DOX, respectively. The drug-loaded cements were tested in an in vitro model of human OS cell line SAOS-2, engineered OS cell line (SAOS-2-eGFP) and U2-OS. The ability of doped scaffolds to induce OS cell death and apoptosis was assessed analysing cell proliferation and Caspase-3/7 activities, respectively. To determine if OS cells grown on doped-scaffolds change their migratory ability and invasiveness, a wound-healing assay was performed. In addition, the osteogenic potential of SrCPC material was evaluated using human adipose derived-mesenchymal stem cells. Osteogenic markers such as (i) the mineral matrix deposition was analysed by alizarin red staining; (ii) the osteocalcin (OCN) protein expression was investigated by enzyme-linked immunosorbent assay test, and (iii) the osteogenic process was studied by real-time polymerase chain reaction array. The delivery system induced cell-killing cytotoxic effects and apoptosis in OS cell lines up to Day 7. SrCPC demonstrates a good cytocompatibility and it induced upregulation of osteogenic genes involved in the skeletal development pathway, together with OCN protein expression and mineral matrix deposition. The proposed approach, based on the local, sustained release of anticancer drugs from nanostructured biomimetic drug-loaded cements is promising for future therapies aiming to combine bone regeneration and anticancer local therapy.
Assuntos
Antineoplásicos , Apoptose , Neoplasias Ósseas , Fosfatos de Cálcio , Doxorrubicina , Metotrexato , Osteogênese , Osteossarcoma , Alicerces Teciduais , Humanos , Antineoplásicos/administração & dosagem , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/patologia , Fosfatos de Cálcio/administração & dosagem , Fosfatos de Cálcio/química , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Doxorrubicina/administração & dosagem , Doxorrubicina/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteossarcoma/tratamento farmacológico , Osteossarcoma/patologia , Osteossarcoma/metabolismo , Estrôncio/farmacologia , Estrôncio/química , Alicerces Teciduais/química , Sistemas de Liberação de Medicamentos , Metotrexato/administração & dosagem , Metotrexato/farmacologiaRESUMO
Bone is a unique type of mineralised connective tissue that can support and protect soft tissues, contain bone marrow, and allow movement [...].
Assuntos
Osso e Ossos , Humanos , Osso e Ossos/metabolismo , Osso e Ossos/fisiologia , AnimaisRESUMO
Bone marrow (BM) microenvironment contributes to the regulation of normal hematopoiesis through a finely tuned balance of self-renewal and differentiation processes, cell-cell interaction, and secretion of cytokines that during leukemogenesis are altered and favor tumor cell growth. In pediatric acute myeloid leukemia (AML), chemotherapy is the standard of care, but >30% of patients still relapse. The need to accelerate the evaluation of innovative medicines prompted us to investigate the role of mesenchymal stromal cells (MSCs) in the leukemic niche to define its contribution to the mechanism of leukemia drug escape. We generated a humanized 3-dimensional (3D) niche with AML cells and MSCs derived from either patients (AML-MSCs) or healthy donors. We observed that AML cells establish physical connections with MSCs, mediating a reprogrammed transcriptome inducing aberrant cell proliferation and differentiation and severely compromising their immunomodulatory capability. We confirmed that AML cells modulate h-MSCs transcriptional profile promoting functions similar to the AML-MSCs when cocultured in vitro, thus facilitating leukemia progression. Conversely, MSCs derived from BM of patients at time of disease remission showed recovered healthy features at transcriptional and functional levels, including the secretome. We proved that AML blasts alter MSCs activities in the BM niche, favoring disease development and progression. We discovered that a novel AML-MSC selective CaV1.2 channel blocker drug, lercanidipine, is able to impair leukemia progression in 3D both in vitro and when implanted in vivo if used in combination with chemotherapy, supporting the hypothesis that synergistic effects can be obtained by dual targeting approaches.
Assuntos
Proliferação de Células , Leucemia Mieloide Aguda/metabolismo , Células-Tronco Mesenquimais/metabolismo , Transcriptoma , Canais de Cálcio Tipo L/metabolismo , Di-Hidropiridinas/farmacologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/patologia , Células-Tronco Mesenquimais/patologia , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/metabolismo , Células Tumorais Cultivadas , Microambiente TumoralRESUMO
Despite the enormous importance of cisplatin as a chemotherapeutic agent, its application is impacted by dose-limiting side effects and lack of selectivity for cancer cells. Researchers can overcome these issues by taking advantage of the pro-drug nature of the platinum(IV) oxidation state, and by modifying the coordination sphere of the metal centre with specific vectors whose receptors are overexpressed in tumour cell membranes (e.g., carbohydrates). In this paper we report the synthesis of four novel carbohydrate-modified Pt(IV) pro-drugs, based on the cisplatin scaffold, and their biological activity against osteosarcoma (OS), a malignant tumour which is most common in adolescents and young adults. The carbohydrate-targeting vectors and Pt scaffold are linked using copper-catalysed azide-alkyne cycloaddition (CuAAC) chemistry, which is synonymous with mild and robust reaction conditions. The novel complexes are characterised using multinuclear 1D-2D NMR (1H, 13C and 195Pt), IR, HR-MS, Elem. Analyses, and CV. Cytotoxicity on 2D and 3D and cell morphology studies on OS cell lines, as well as non-cancerous human foetal osteoblasts (hFOBs), are discussed.
Assuntos
Antineoplásicos , Neoplasias Ósseas , Complexos de Coordenação , Osteossarcoma , Pró-Fármacos , Humanos , Adolescente , Cisplatino/uso terapêutico , Linhagem Celular Tumoral , Antineoplásicos/química , Osteossarcoma/tratamento farmacológico , Osteossarcoma/patologia , Platina/química , Pró-Fármacos/química , Complexos de Coordenação/química , Neoplasias Ósseas/tratamento farmacológico , CarboidratosRESUMO
This work describes the development of an injectable nanocomposite system based on a chitosan thermosensitive hydrogel combined with liposomes for regenerative medicine applications. Liposomes with good physicochemical properties are prepared and embedded within the chitosan network. The resulting nanocomposite hydrogel is able to provide a controlled release of the content from liposomes, which are able to interact with cells and be internalized. The cellular uptake is enhanced by the presence of a chitosan coating, and cells incubated with liposomes embedded within thermosensitive hydrogels displayed a higher cell uptake compared to cells incubated with liposomes alone. Furthermore, the gelation temperature of the system resulted to be equal to 32.6 °C; thus, the system can be easily injected in the target site to form a hydrogel at physiological temperature. Given the peculiar performance of the selected systems, the resulting thermosensitive hydrogels are a versatile platform and display potential applications as controlled delivery systems of liposomes for tissue regeneration.
Assuntos
Quitosana , Portadores de Fármacos , Sistemas de Liberação de Medicamentos , Hidrogéis , Lipossomos , Medicina Regenerativa , Temperatura , Animais , Linhagem Celular , Fenômenos Químicos , Quitosana/química , Portadores de Fármacos/química , Humanos , Hidrogéis/química , Lipossomos/química , Camundongos , Medicina Regenerativa/métodosRESUMO
The aging of the world population is increasingly claimed as an alarming situation, since an ever-raising number of persons in advanced age but still physically active is expected to suffer from invalidating and degenerative diseases. The impairment of the endogenous healing potential provoked by the aging requires the development of more effective and personalized therapies, based on new biomaterials and devices able to direct the cell fate to stimulate and sustain the regrowth of damaged or diseased tissues. To obtain satisfactory results, also in cases where the cell senescence, typical of the elderly, makes the regeneration process harder and longer, the new solutions have to possess excellent ability to mimic the physiological extracellular environment and thus exert biomimetic stimuli on stem cells. To this purpose, the "biomimetic concept" is today recognized as elective to fabricate bioactive and bioresorbable devices such as hybrid osteochondral scaffolds and bioactive bone cements closely resembling the natural hard tissues and with enhanced regenerative ability. The review will illustrate some recent results related to these new biomimetic materials developed for application in different districts of the musculoskeletal system, namely bony, osteochondral and periodontal regions, and the spine. Further, it will be shown how new bioactive and superparamagnetic calcium phosphate nanoparticles can give enhanced results in cardiac regeneration and cancer therapy. Since tissue regeneration will be a major demand in the incoming decades, the high potential of biomimetic materials and devices is promising to significantly increase the healing rate and improve the clinical outcomes even in aged patients.
Assuntos
Materiais Biomiméticos , Alicerces Teciduais , Idoso , Humanos , Engenharia TecidualRESUMO
Biomaterial science increasingly seeks more biomimetic scaffolds that functionally augment the native bone tissue. In this paper, a new concept of a structural scaffold design is presented where the physiological multi-scale architecture is fully incorporated in a single-scaffold solution. Hydroxyapatite (HA) and ß-tricalcium phosphate (ß-TCP) bioceramic scaffolds with different bioinspired porosity, mimicking the spongy and cortical bone tissue, were studied. In vitro experiments, looking at the mesenchymal stem cells behaviour, were conducted in a perfusion bioreactor that mimics the physiological conditions in terms of interstitial fluid flow and associated induced shear stress. All the biomaterials enhanced cell adhesion and cell viability. Cortical bone scaffolds, with an aligned architecture, induced an overexpression of several late stage genes involved in the process of osteogenic differentiation compared to the spongy bone scaffolds. This study reveals the exciting prospect of bioinspired porous designed ceramic scaffolds that combines both cortical and cancellous bone in a single ceramic bone graft. It is prospected that dual core shell scaffold could significantly modulate osteogenic processes, once implanted in patients, rapidly forming mature bone tissue at the tissue interface, followed by subsequent bone maturation in the inner spongy structure.
Assuntos
Materiais Biocompatíveis/química , Osso e Ossos/metabolismo , Células-Tronco/citologia , Tecido Adiposo , Animais , Reatores Biológicos , Fosfatos de Cálcio/química , Diferenciação Celular , Sobrevivência Celular , Células Cultivadas , Cerâmica/química , Durapatita/química , Líquido Extracelular , Humanos , Técnicas In Vitro , Células-Tronco Mesenquimais/citologia , Microscopia Eletrônica de Varredura , Osteogênese , Polímeros/química , Porosidade , Pós , Alicerces Teciduais/químicaRESUMO
A hot topic in biomedical science is the implementation of more predictive in vitro models of human tissues to significantly improve the knowledge of physiological or pathological process, drugs discovery and screening. Bidimensional (2D) culture systems still represent good high-throughput options for basic research. Unfortunately, these systems are not able to recapitulate the in vivo three-dimensional (3D) environment of native tissues, resulting in a poor in vitro-in vivo translation. In addition, intra-species differences limited the use of animal data for predicting human responses, increasing in vivo preclinical failures and ethical concerns. Dealing with these challenges, in vitro 3D technological approaches were recently bioengineered as promising platforms able to closely capture the complexity of in vivo normal/pathological tissues. Potentially, such systems could resemble tissue-specific extracellular matrix (ECM), cell-cell and cell-ECM interactions and specific cell biological responses to mechanical and physical/chemical properties of the matrix. In this context, this review presents the state of the art of the most advanced progresses of the last years. A special attention to the emerging technologies for the development of human 3D disease-relevant and physiological models, varying from cell self-assembly (i.e., multicellular spheroids and organoids) to the use of biomaterials and microfluidic devices has been given.
Assuntos
Tecnologia Biomédica , Técnicas de Cultura de Células , Corpo Humano , Modelos Biológicos , Esferoides Celulares , Animais , Materiais Biocompatíveis/química , Bioimpressão , Microfluídica/métodos , Nanotecnologia , Organoides , Técnicas de Cultura de Tecidos , Engenharia TecidualRESUMO
A new straightforward gel permeation chromatography (GPC) method was developed to calculate the drug encapsulation efficiency and loading content of Poly(lactic acid) nanoparticles (PLA NPs) loaded with Salinomycin (Sal), exploiting the capability of this technique to separate a macromolecular/molecular mixture on the basis of the molecular weight of each component. The proposed GPC method allowed Sal detection until 1% of Sal content in PLA NPs, avoiding sample pre-treatments. The method was validated by wave voltammetry (SW) technique, using a slightly modified literature procedure, useful to detect Sal in the concentration range 0.4 ≤ C/µmol/L ≤ 12 (linear concentration range). PLA-based NPs were prepared by nanoprecipitation with either native and functionalized PLA. Specifically, folate-decorated PLA NPs (PLA-FA NPs) were obtained by CuAAC click functionalization of alkyne-grafted PLA with azide-folate. Sal-loaded NPs were characterized physicochemically and morphologically. They exhibited adequate physicochemical properties, good drug encapsulation efficiency (98 ± 0.5% and 99 ± 0.5%), and loading content (8.8 ± 0.1% and 8.9 ± 0.1% for PLA/Sal and PLA-FA/Sal NPs, respectively). The size of empty PLA NPs resulted smaller (90 ± 3.2 nm and 680 ± 15.3 nm, for PLA NPs and PLA-FA NPs respectively) than the correspondent drug-loaded NPs (110 ± 3.8 nm and 875 ± 20.5 nm, respectively). Their biological activity was assessed on osteosarcoma bulk cells MG63, healthy osteoblast cell line (hFOB1.19), and enriched osteosarcoma cancer stem cells (CSCs), showing cell-depending effect. Entrapped Sal maintained its cytotoxic effect on CSCs and MG63 cells, with a potency comparable to the free drug and no evident benefit was detected for folate-decorated PLA NPs respect to native PLA NPs. Graphical abstract.
Assuntos
Antineoplásicos/administração & dosagem , Portadores de Fármacos/química , Nanopartículas/química , Poliésteres/química , Piranos/administração & dosagem , Antineoplásicos/análise , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Neoplasias Ósseas/tratamento farmacológico , Linhagem Celular Tumoral , Cromatografia/métodos , Humanos , Osteossarcoma/tratamento farmacológico , Piranos/análise , Piranos/farmacocinética , Piranos/farmacologiaRESUMO
Many medical-related scientific discoveries arise from trial-error patterns where the processes involved must be refined and modified continuously before any product could be able to reach the final costumers. One of the elements affecting negatively these processes is the inaccuracy of two-dimension (2D) standard culture systems, carried over in plastic plates or similar, in replicating complex environments and patterns. Consequently, animal tests are required to validate every in vitro finding, at the expenses of more funds and ethical issues. A possible solution relies in the implementation of three-dimension (3D) culture systems as a fitting gear between the 2D tests and in vivo tests, aiming to reduce the negative in vivo outcomes. These 3D structures are depending from the comprehension of the extracellular matrix (ECM) and the ability to replicate it in vitro. In this article a comparison of efficacies between these two culture systems was taken as subject, human mesenchymal stem cells (hMSCs) was utilized and a hybrid scaffold made by a blend of chitosan, gelatin and biomineralized gelatin was used for the 3D culture system.
Assuntos
Técnicas de Cultura de Células , Células-Tronco Mesenquimais/fisiologia , Osteogênese/fisiologia , Alicerces Teciduais , Materiais Biocompatíveis , Diferenciação Celular , Humanos , Teste de MateriaisRESUMO
Nanocrystalline apatites mimicking bone mineral represent a versatile platform for biomedical applications thanks to their similarity to bone apatite and the possibility to (multi)functionalize them so as to provide "à la carte" properties. One relevant domain is in particular oncology, where drug-loaded biomaterials and engineered nanosystems may be used for diagnosis, therapy, or both. In a previous contribution, we investigated the adsorption of doxorubicin onto two nanocrystalline apatite substrates, denoted HA and FeHA (superparamagnetic apatite doped with iron ions), and explored these drug-loaded systems against tumor cells. To widen their applicability in the oncology field, here we examine the interaction between the same two substrates and two other molecules: folic acid (FA), often used as cell targeting agent, and the anticancer drug methotrexate (MTX), an antifolate analogue. In a first stage, we investigated the adsorptive behavior of FA (or MTX) on both substrates, evidencing their specificities. At low concentration, typically under 100 mmol/L, adsorption onto HA was best described using the Sips isotherm model, while the formation of a calcium folate secondary salt was evidenced at high concentration by Raman spectroscopy. Adsorption onto FeHA was instead fitted to the Langmuir model. A larger adsorptive affinity was found for the FeHA substrate compared to HA; accordingly, a faster release was noticed from HA. In vitro tests carried out on human osteosarcoma cell line (SAOS-2) allowed us to evaluate the potential of these compounds in oncology. Finally, in vivo (subcutaneous) implantations in the mouse were run to ascertain the biocompatibility of the two substrates. These results should allow a better understanding of the interactions between FA/MTX and bioinspired nanocrystalline apatites in view of applications in the field of cancer.
Assuntos
Antineoplásicos/farmacologia , Antagonistas do Ácido Fólico/farmacologia , Ácido Fólico/química , Hidroxiapatitas/química , Metotrexato/farmacologia , Adsorção , Animais , Antineoplásicos/química , Materiais Biocompatíveis/química , Materiais Biocompatíveis/toxicidade , Linhagem Celular Tumoral , Liberação Controlada de Fármacos , Antagonistas do Ácido Fólico/química , Humanos , Hidroxiapatitas/toxicidade , Metotrexato/química , Camundongos Endogâmicos C57BL , Nanopartículas/química , Nanopartículas/toxicidadeRESUMO
Doping of biocompatible nanomaterials with magnetic phases is currently one of the most promising strategies for the development of advanced magnetic biomaterials. However, especially in the case of iron-doped magnetic hydroxyapatites, it is not clear if the magnetic features come merely from the magnetic phases/ions used as dopants or from complex mechanisms involving interactions at the nanoscale. Here, we report an extensive chemical-physical and magnetic investigation of three hydroxyapatite nanocrystals doped with different iron species and containing small or no amounts of maghemite as a secondary phase. The association of several investigation techniques such as X-ray absorption spectroscopy, Mössbauer, magnetometry, and TEM allowed us to determine that the unusual magnetic properties of Fe2+/3+-doped hydroxyapatites (FeHA) occur by a synergy of two different phenomena: i.e., (i) interacting superparamagnetism due to the interplay between iron-doped apatite and iron oxide nanoparticles as well as to the occurrence of dipolar interactions and (ii) interacting paramagnetism due to Fe3+ ions present in the superficial hydrated layer of the apatite nanophase and, to a lesser extent, paramagnetism due to isolated Fe3+ ions in the apatite lattice. We also show that a major player in the activation of the above phenomena is the oxidation of Fe2+ into Fe3+, as induced by the synthesis process, and their consequent specific positioning in the FeHA structure.
Assuntos
Hidroxiapatitas/química , Ferro/química , Fenômenos Magnéticos , Nanopartículas/química , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
Understanding the mineralization mechanism of synthetic protein has recently aroused great interest especially in the development of advanced materials for bone regeneration. Herein, we propose the synthesis of composite materials through the mineralization of a recombinant collagen type I derived protein (RCP) enriched with RGD sequences in the presence of magnesium ions (Mg) to closer mimic bone composition. The role of both RCP and Mg ions in controlling the precipitation of the mineral phase is in depth evaluated. TEM and X-ray powder diffraction reveal the crystallization of nanocrystalline apatite (Ap) in all the evaluated conditions. However, Raman spectra point out also the precipitation of amorphous calcium phosphate (ACP). This amorphous phase is more evident when RCP and Mg are at work, indicating the synergistic role of both in stabilizing the amorphous precursor. In addition, hybrid matrices are prepared to tentatively address their effectiveness as scaffolds for bone tissue engineering. SEM and AFM imaging show an homogeneous mineral distribution on the RCP matrix mineralized in presence of Mg, which provides a surface roughness similar to that found in bone. Preliminary in vitro tests with pre-osteoblast cell line show good cell-material interaction on the matrices prepared in the presence of Mg. To the best of our knowledge this work represents the first attempt to mineralize recombinant collagen type I derived protein proving the simultaneous effect of the organic phase (RCP) and Mg on ACP stabilization. This study opens the possibility to engineer, through biomineralization process, advanced hybrid matrices for bone regeneration.
Assuntos
Regeneração Óssea , Calcificação Fisiológica , Engenharia Tecidual/métodos , Animais , Apatitas , Biomimética/métodos , Linhagem Celular , Colágeno Tipo I/metabolismo , Magnésio , Camundongos , Minerais , Engenharia de ProteínasRESUMO
New magnetic hydroxyapatite-based nanomaterials as bone-specific systems for controlled drug delivery have been synthesized. The synthesized hydroxyapatite, HA, decorated with magnetite nanoparticles by a deposition method (HA/Fe3O4) and the nanocomposite system obtained using magnetic multi-walled carbon nanotubes (HA/MWCNT/Fe3O4) as a filler for HA have been characterized by chemical and morphological analyses, and their biological behavior was investigated. The systems have also been doped with clodronate in order to combine the effect of bone biomineralization induced by hydroxyapatite-based composites with the decrease of osteoclast formation induced by the drug. An analysis of the preosteoclastic RAW264.7 cell proliferation by MTT assay confirmed the high biocompatibility of the three systems. TRAP staining of RAW 264.7 conditioned with sRAKL to induce osteoclastogenesis, cultured in the presence of the systems doped and undoped with clodronate, showed the inhibitory effect of clodronate after we counted the MNC TRAP(+)cells but only in the osteoclast formation; in particular, the system HA/Fe3O4-Clo exerted a high inhibitory effect compared to the drug alone. These results demonstrate that the synthesized nanocomposites are a biocompatible magnetic drug delivery system and can represent a useful multimodal platform for applications in bone tissue engineering.
Assuntos
Materiais Biocompatíveis/síntese química , Sistemas de Liberação de Medicamentos/instrumentação , Durapatita/síntese química , Nanopartículas de Magnetita/química , Nanotubos de Carbono/química , Osteoclastos/efeitos dos fármacos , Engenharia Tecidual/instrumentação , Animais , Osso e Ossos/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sistemas de Liberação de Medicamentos/métodos , Camundongos , Tamanho da Partícula , Engenharia Tecidual/métodosRESUMO
Collagen electrospun scaffolds well reproduce the structure of the extracellular matrix (ECM) of natural tissues by coupling high biomimetism of the biological material with the fibrous morphology of the protein. Structural properties of collagen electrospun fibers are still a debated subject and there are conflicting reports in the literature addressing the presence of ultrastructure of collagen in electrospun fibers. In this work collagen type I was successfully electrospun from two different solvents, trifluoroethanol (TFE) and dilute acetic acid (AcOH). Characterization of collagen fibers was performed by means of SEM, ATR-IR, Circular Dichroism and WAXD. We demonstrated that collagen fibers contained a very low amount of triple helix with respect to pristine collagen (18 and 16% in fibers electrospun from AcOH and TFE, respectively) and that triple helix denaturation occurred during polymer dissolution. Collagen scaffolds were crosslinked by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), a commonly employed crosslinker for electrospun collagen, and 1,4-butanediol diglycidyl ether (BDDGE), that was tested for the first time in this work as crosslinking agent for collagen in the form of electrospun fibers. We demonstrated that BDDGE successfully crosslinked collagen and preserved at the same time the scaffold fibrous morphology, while scaffolds crosslinked with EDC completely lost their porous structure. Mesenchymal stem cell experiments demonstrated that collagen scaffolds crosslinked with BDDGE are biocompatible and support cell attachment.
Assuntos
Colágeno/química , Reagentes de Ligações Cruzadas/farmacologia , Nanofibras/química , Solventes/farmacologia , Alicerces Teciduais , Animais , Materiais Biocompatíveis/química , Butileno Glicóis , Células Cultivadas , Colágeno/efeitos dos fármacos , Estabilidade de Medicamentos , Galvanoplastia/métodos , Matriz Extracelular/química , Teste de Materiais , Coelhos , Engenharia Tecidual/métodos , Alicerces Teciduais/químicaRESUMO
Shape memory alloys based on NiTi have found their main applications in manufacturing of new biomedical devices mainly in surgery tools, stents and orthopedics. Porous NiTi can exhibit an engineering elastic modulus comparable to that of cortical bone (12-17 GPa). This condition, combined with proper pore size, allows good osteointegration. Open cells porous NiTi was produced by self propagating high temperature synthesis (SHS), starting from Ni and Ti mixed powders. The main NiTi phase is formed during SHS together with other Ni-Ti compounds. The biocompatibility of such material was investigated by single culture experiment and ionic release on small specimen. In particular, NiTi and porous NiTi were evaluated together with elemental Ti and Ni reference metals and the two intermetallic TiNi3, Ti2Ni phases. This approach permitted to clearly identify the influence of secondary phases in porous NiTi materials and relation with Ni-ion release. The results indicated, apart the well-known high toxicity of Ni, also toxicity of TiNi3, whilst phases with higher Ti content showed high biocompatibility. A slightly reduced biocompatibility of porous NiTi was ascribed to combined effect of TiNi3 presence and topography that requires higher effort for the cells to adapt to the surface.
Assuntos
Ligas/síntese química , Materiais Biocompatíveis/química , Temperatura Alta , Níquel/química , Polimerização , Titânio/química , Ligas/química , Ligas/farmacologia , Células Cultivadas , Humanos , Teste de Materiais , Níquel/farmacologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/fisiologia , Porosidade , Pós , Propriedades de Superfície , Titânio/farmacologiaRESUMO
A series of C2-functionalied Pt (IV) glycoconjugates based on glucosamine have been synthesised, characterised and tested as anticancer agents on a series of different 2D and 3D cancer cell lines. The carbohydrate will act as a targeted delivery system to improve the selectivity, exploiting the Warburg Effect and the GLUTs receptors that are overexpressed in most of the cancer cells. The hydroxyl at C2 of the carbohydrates does not participate in hydrogen bonding with the GLUTs receptors, making C2 an attractive position for drug conjugation as seen in literature. In this study, we use the amino functionality at the C2 position in glucosamine and Copper-catalysed Azide-Alkyne Cycloaddition "click" (CuAAC) reaction to connect the prodrug Pt (IV) scaffold to the carbohydrate. We have investigated complexes with different linker lengths, as well as acetyl protected and free derivatives. To the best of our knowledge, this study represents the first series of Pt (IV) glucosamine-conjugates functionalised at C2.
RESUMO
We developed cyclic RGD-tagged polymeric micellar nanoassemblies for sustained delivery of Doxorubicin (Dox) endowed with significant cytotoxic effect against MG63, SAOS-2, and U2-OS osteosarcoma cells without compromising the viability of healthy osteoblasts (hFOBs). Targeted polymeric micellar nanoassemblies (RGD-NanoStar@Dox) enabled Dox to reach the nucleus of MG63, SAOS-2, and U2-OS cells causing the same cytotoxic effect as free Dox, unlike untargeted micellar nanoassemblies (NanoStar@Dox) which failed to reach the nucleus and resulted ineffective, demonstrating the crucial role of cyclic RGD peptide in driving cellular uptake and accumulation mechanisms in osteosarcoma cells. Micellar nanoassemblies were obtained by nanoformulation of three-armed star PLA-PEG copolymers properly synthetized with and without decoration with the cyclic-RGDyK peptide (Arg-Gly-Asp-D-Tyr-Lys). The optimal RGD-NanoStar@Dox nanoformulation obtained by nanoprecipitation method (8 % drug loading; 35 % encapsulation efficiency) provided a prolonged and sustained drug release with a rate significantly lower than the free drug under the same experimental conditions. Moreover, the nanosystem preserved Dox from the natural degradation occurring under physiological conditions (i.e., dimerization and consequent precipitation) serving as a slow-release "drug reservoir" ensuring an extended biological activity over the time.
Assuntos
Neoplasias Ósseas , Sobrevivência Celular , Doxorrubicina , Micelas , Oligopeptídeos , Osteossarcoma , Polietilenoglicóis , Doxorrubicina/administração & dosagem , Doxorrubicina/farmacologia , Doxorrubicina/química , Osteossarcoma/tratamento farmacológico , Humanos , Polietilenoglicóis/química , Linhagem Celular Tumoral , Oligopeptídeos/química , Oligopeptídeos/administração & dosagem , Neoplasias Ósseas/tratamento farmacológico , Sobrevivência Celular/efeitos dos fármacos , Nanopartículas/química , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/farmacologia , Antibióticos Antineoplásicos/química , Liberação Controlada de Fármacos , Portadores de Fármacos/químicaRESUMO
Cellular alignment plays a pivotal role in several human tissues, including skeletal muscle, spinal cord and tendon. Various techniques have been developed to control cellular alignment using 3D biomaterials. However, the majority of 3D-aligned scaffolds require invasive surgery for implantation. In contrast, injectable hydrogels provide a non-invasive delivery method, gaining considerable attention for the treatment of diverse conditions, including osteochondral lesions, volumetric muscle loss, and traumatic brain injury. We engineered a biomimetic hydrogel with magnetic responsiveness by combining gellan gum, hyaluronic acid, collagen, and magnetic nanoparticles (MNPs). Collagen type I was paired with MNPs to form magnetic collagen bundles (MCollB), allowing the orientation control of these bundles within the hydrogel matrix through the application of a remote low-intensity magnetic field. This resulted in the creation of an anisotropic architecture. The hydrogel mechanical properties were comparable to those of human soft tissues, such as skeletal muscle, and proof of the aligned hydrogel concept was demonstrated. In vitro findings confirmed the absence of toxicity and pro-inflammatory effects. Notably, an increased fibroblast cell proliferation and pro-regenerative activation of macrophages were observed. The in-vivo study further validated the hydrogel biocompatibility and demonstrated the feasibility of injection with rapid in situ gelation. Consequently, this magnetically controlled injectable hydrogel exhibits significant promise as a minimally invasive, rapid gelling and effective treatment for regenerating various aligned human tissues.
RESUMO
The global diffusion of antibiotic resistance poses a severe threat to public health. Addressing antibiotic-resistant infections requires innovative approaches, such as antibacterial nanostructured surfaces (ANSs). These surfaces, featuring ordered arrays of nanostructures, exhibit the ability to kill bacteria upon contact. However, most currently developed ANSs utilize bioinert materials, lacking bioactivity crucial for promoting tissue regeneration, particularly in the context of bone infections. This study introduces ANSs composed of bioactive calcium phosphate nanocrystals. Two distinct ANSs were created through a biomineralization-inspired growth of amorphous calcium phosphate (ACP) precursors. The ANSs demonstrated efficient antibacterial properties against both Gram-negative (P. aeruginosa) and Gram-positive (S. aureus) antibiotic resistant bacteria, with up to 75 % mortality in adhered bacteria after only 4 h of contact. Notably, the ANS featuring thinner and less oriented nano-needles exhibited superior efficacy attributed to simultaneous membrane rupturing and oxidative stress induction. Moreover, the ANSs facilitate the proliferation of mammalian cells, enhancing adhesion, spreading, and reducing oxidative stress. The ANSs displayed also significant bioactivity towards human mesenchymal stem cells, promoting colonization and inducing osteogenic differentiation. Specifically, the ANS with thicker and more ordered nano-needles demonstrated heightened effects. In conclusion, ANSs introduced in this work have the potential to serve as foundation for developing bone graft materials capable of eradicate site infections while concurrently stimulating bone regeneration. STATEMENT OF SIGNIFICANCE: Nanostructured surfaces with antibacterial properties through a mechano-bactericidal mechanism have shown significant potential in fighting antibiotic resistance. However, these surfaces have not been fabricated with bioactive materials necessary for developing devices that are both antibacterial and able to stimulate tissue regeneration. This study demonstrates the feasibility of creating nanostructured surfaces of ordered calcium phosphate nano-needles through a biomineralization-inspired growth. These surfaces exhibit dual functionality, serving as effective bactericidal agents against Gram-negative and Gram-positive antibiotic-resistant bacteria while also promoting the proliferation of mammalian cells and inducing osteogenic differentiation of human mesenchymal stem cells. Consequently, this approach holds promise in the context of bone infections, introducing innovative nanostructured surfaces that could be utilized in the development of antimicrobial and osteogenic grafts.