RESUMO
BACKGROUND BURKHOLDERIA: is a phosphorus solubilizing microorganism discovered in recent years, which can dissolve insoluble phosphorus compounds into soluble phosphorus. To investigate the effects of Burkholderia and calcium phosphate on the composting of Torreya grandis branches and leaves, as well as to explain the nutritional and metabolic markers related to the composting process. METHODS: In this study, we employed amplicon sequencing and untargeted metabolomics analysis to examine the interplay among phosphorus (P) components, microbial communities, and metabolites during T. grandis branch and leaf waste composting that underwent treatment with calcium phosphate and phosphate-solubilizing bacteria (Burkholderia). There were four composting treatments, 10% calcium phosphate (CaP) or 5 ml/kg (1 × 108/ml Burkholderia) microbial inoculum (WJP) or both (CaP + WJP), and the control group (CK). RESULTS: The results indicated that Burkholderia inoculation and calcium phosphate treatment affected the phosphorus composition, pH, EC, and nitrogen content. Furthermore, these treatments significantly affected the diversity and structure of bacterial and fungal communities, altering microbial and metabolite interactions. The differential metabolites associated with lipids and organic acids and derivatives treated with calcium phosphate treatment are twice as high as those treated with Burkholderia in both 21d and 42d. The results suggest that calcium phosphate treatment alters the formation of some biological macromolecules. CONCLUSION: Both Burkholderia inoculation and calcium phosphate treatment affected the phosphorus composition, nitrogen content and metabolites of T. grandis branch and leaf waste compost.These results extend our comprehension of the coupling of matter transformation and community succession in composting with the addition of calcium phosphate and phosphate-solubilizing bacteria.
Assuntos
Burkholderia , Fosfatos de Cálcio , Compostagem , Fósforo , Microbiologia do Solo , Fosfatos de Cálcio/metabolismo , Fósforo/metabolismo , Burkholderia/metabolismo , Burkholderia/genética , Burkholderia/efeitos dos fármacos , Bactérias/metabolismo , Bactérias/genética , Bactérias/classificação , Bactérias/efeitos dos fármacos , Microbiota/efeitos dos fármacos , Nitrogênio/metabolismo , Solo/química , Folhas de Planta/microbiologia , Fungos/metabolismo , Fungos/efeitos dos fármacos , Fungos/genética , Fungos/classificação , Concentração de Íons de HidrogênioRESUMO
BACKGROUND: Tyrosine-rich amelogenin peptide (TRAP) is the main amelogenin digestion product in the developmental enamel matrix. It has been shown to promote remineralization of demineralized enamel in our previous study. However, direct evidence of the effect of TRAP on the morphology and nanostructure of crystal growth on an enamel surface has not been reported. This study aimed to examine the effect of TRAP on the morphology of calcium phosphate crystals grown on early enamel erosion using a pH-cycling model. METHODS: Eroded lesions were produced in human premolars by 30-second immersion in 37% phosphoric acid. Forty-five samples of eroded human premolar enamel blocks were selected and randomly divided into 3 groups: deionized water (DDW, negative control); 100 µg/mL TRAP, and 2 ppm sodium fluoride (NaF, positive control group). For 14 days, the specimens were exposed to a pH-cycling model. Using scanning electron microscopy (SEM) and atomic force microscopy (AFM) methods, the surface morphology, calcium-phosphorus ratio, and enamel surface roughness were examined. X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR) were used to assess crystal characteristics. RESULTS: After pH-cycling, compared to the two control groups, the surface of the eroded enamel of the peptide TRAP group shows a large number of new, densely arranged rod-like crystals, parallel to each other, regularly arranged, forming an ordered structure, with crystal morphology similar to that of natural enamel. The crystals are mostly hydroxyapatite (HA). CONCLUSION: This study demonstrates that the peptide TRAP modulates the formation of hydroxyapatite in eroded enamel and that the newly formed crystals resemble natural enamel crystals and promote the remineralization of enamel, providing a promising biomaterial for remineralization treatment of enamel lesions.
Assuntos
Amelogenina , Esmalte Dentário , Microscopia Eletrônica de Varredura , Erosão Dentária , Remineralização Dentária , Difração de Raios X , Humanos , Remineralização Dentária/métodos , Esmalte Dentário/efeitos dos fármacos , Erosão Dentária/patologia , Concentração de Íons de Hidrogênio , Amelogenina/uso terapêutico , Amelogenina/farmacologia , Espectroscopia de Infravermelho com Transformada de Fourier , Microscopia de Força Atômica , Fosfatos de Cálcio/farmacologia , Propriedades de Superfície , Dente Pré-Molar , CristalizaçãoRESUMO
Gene therapy using a protein-based CRISPR system in the brain has practical limitations due to current delivery systems, especially in the presence of arterial occlusion. To overcome these obstacles and improve stability, we designed a system for intranasal administration of gene therapy for the treatment of ischemic stroke. Methods: Nanoparticles containing the protein-based CRISPR/dCas9 system targeting Sirt1 were delivered intranasally to the brain in a mouse model of ischemic stroke. The CRISPR/dCas9 system was encapsulated with calcium phosphate (CaP) nanoparticles to prevent them from being degraded. They were then conjugated with ß-hydroxybutyrates (bHb) to target monocarboxylic acid transporter 1 (MCT1) in nasal epithelial cells to facilitate their transfer into the brain. Results: Human nasal epithelial cells were shown to uptake and transfer nanoparticles to human brain endothelial cells with high efficiency in vitro. The intranasal administration of the dCas9/CaP/PEI-PEG-bHb nanoparticles in mice effectively upregulated the target gene, Sirt1, in the brain, decreased cerebral edema and increased survival after permanent middle cerebral artery occlusion. Additionally, we observed no significant in vivo toxicity associated with intranasal administration of the nanoparticles, highlighting the safety of this approach. Conclusion: This study demonstrates that the proposed protein-based CRISPR-dCas9 system targeting neuroprotective genes in general, and SIRT1 in particular, can be a potential novel therapy for acute ischemic stroke.
Assuntos
Administração Intranasal , Encéfalo , Modelos Animais de Doenças , Terapia Genética , AVC Isquêmico , Nanopartículas , Sirtuína 1 , Animais , Camundongos , Humanos , AVC Isquêmico/terapia , AVC Isquêmico/genética , Nanopartículas/administração & dosagem , Terapia Genética/métodos , Sirtuína 1/genética , Sirtuína 1/metabolismo , Encéfalo/metabolismo , Masculino , Fosfatos de Cálcio , Sistemas CRISPR-Cas , Camundongos Endogâmicos C57BL , Células Endoteliais/metabolismo , Isquemia Encefálica/terapia , Isquemia Encefálica/genética , Infarto da Artéria Cerebral Média/terapia , Infarto da Artéria Cerebral Média/genética , Células Epiteliais/metabolismoRESUMO
The development of synthetic bone substitutes that equal or exceed the efficacy of autologous graft remains challenging. In this study, a rat calvarial defect model was used as a reference to investigate the influence of composition and architecture of 3D-printed cement, with or without bioactives, on tissue regeneration. Printable cement pastes were formulated by combining hyaluronic acid and cement precursors. Cementitious scaffolds were printed with 3 different patterns. After 7 weeks of implantation with or without bone marrow, multiparametric qualitative and quantitative assessments were performed using µCT, SEM, and histology. None of the set-up strategies was as efficient as autologous cancellous bone graft to repair calvarial defects. Nonetheless, the presence of scaffold improved the skull vault closure, particularly when the scaffold was soaked in total bone marrow before implantation. No significant effect of scaffold macro-architecture was observed on tissue mineralization. Magnesium phosphate-based scaffolds (MgP) seemed to induce higher bone formation than their calcium-phosphate-based counterparts. They also displayed a quicker biodegradation and sparse remaining material was found after 7 weeks of implantation. Although further improvements are required to reach clinical settings, this study demonstrated the potential of organo-mineral cements for bone regeneration and highlighted the peculiar properties of MgP-based cements.
Assuntos
Regeneração Óssea , Impressão Tridimensional , Crânio , Alicerces Teciduais , Animais , Regeneração Óssea/efeitos dos fármacos , Alicerces Teciduais/química , Ratos , Crânio/efeitos dos fármacos , Substitutos Ósseos/química , Substitutos Ósseos/farmacologia , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Masculino , Cimentos Ósseos/farmacologia , Cimentos Ósseos/química , Fosfatos/química , Osteogênese/efeitos dos fármacos , Microtomografia por Raio-X , Compostos de MagnésioRESUMO
Premixed calcium silicate-based materials have recently been developed and are recommended for a wide range of endodontic procedures, including vital pulp therapy. This study investigated the in vitro biocompatibility and pro-mineralization effect and in vivo reparative dentin formation of EndoSequence Root Repair Material, EndoSequence BCRRM, Bio-C Repair, and Well-pulp PT. Both fresh and set extracts had no detrimental effect on the growth of human dental pulp stem cells. The fresh extracts had a higher calcium concentration than the set extracts and induced considerably greater mineralized nodule formation. EndoSequence Root Repair Material had the longest setting time, whereas Bio-C Repair had the shortest. When these materials were applied to exposed rat molar pulps, mineralized tissue deposition was found at the exposure sites after 2 weeks. These results indicate that the premixed calcium silicate-based materials tested could have positive benefits for direct pulp capping procedures.
Assuntos
Materiais Biocompatíveis , Compostos de Cálcio , Polpa Dentária , Silicatos , Células-Tronco , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/citologia , Silicatos/farmacologia , Compostos de Cálcio/farmacologia , Humanos , Células-Tronco/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Ratos , Animais , Teste de Materiais , Células Cultivadas , Técnicas In Vitro , Masculino , Fosfatos de Cálcio , Combinação de Medicamentos , ÓxidosRESUMO
Background: The most problematic complication of external fixation is infection at the pin insertion site. Technology that improves the adhesion of the external fixation pin to the skin, subcutaneous tissue, and bone may prevent infection at the pin site. The purpose of this study is to formulate a calcium phosphate-fibroblast growth factor (Cp-FGF) coating on a stainless-steel external fixation pin and to verify its effectiveness in reducing infection at the pin site and its possible influence on bone fixation in animal experiments. Methods: We compared stainless-steel screws without coating (SS group; n = 32), those with a calcium phosphate coating (Cp group; n = 30), those with a Cp-FGF coating (FGF group; n = 32), and those with a Cp-FGF coating having enhanced biological activity (FGF+ group; n = 32) in male Japanese white domesticated rabbits. Screws were inserted percutaneously into the bilateral proximal tibial diaphysis of the rabbits and implanted for 4 weeks. Screws and periscrew tissue were observed postoperatively for qualitatively assessing infection. Results: Infection assessment by gross findings after 4 weeks (at screw removal) showed no significant differences between the groups. Histopathological evaluation of soft tissue infection and bone tissue infection showed no significant differences between the groups for either soft tissue or bone tissue. Since neither the FGF+ group nor the FGF group showed anti-infective effects, the biological activity of FGF is not the only determining factor. We compared SEM, XRD, coating detaching test, sustained release test, and bioassay to examine physicochemical properties among the coatings but found no sufficient differences. Conclusions: It is suggested that improving the tissue adhesion to and/or biocompatibility of pins is also important to improve the in vivo performance of Cp-FGF-coated external fixation pins.
Assuntos
Fatores de Crescimento de Fibroblastos , Aço Inoxidável , Animais , Coelhos , Masculino , Fatores de Crescimento de Fibroblastos/uso terapêutico , Fatores de Crescimento de Fibroblastos/administração & dosagem , Fatores de Crescimento de Fibroblastos/farmacologia , Pinos Ortopédicos , Materiais Revestidos Biocompatíveis , Fosfatos de Cálcio/uso terapêutico , Anti-Infecciosos/farmacologia , Anti-Infecciosos/uso terapêutico , Anti-Infecciosos/administração & dosagemRESUMO
This study evaluated push-out bond test (POBT), surface roughness, and antimicrobial properties against Enterococcus faecalis of bioceramic sealers supplemented with silver nanoparticles (AgNPs). The sealers tested were CeraSeal®, EndoSequence® BC SealerTM, and Bio-C® Sealer. The POBT was measured with a Universal Testing Machine, and the type of failure was evaluated with a stereomicroscope. The roughness average (Sa) and peak-valley height (Sy) values were evaluated by atomic force microscopy. The bacterial growth inhibition was evaluated using a disk diffusion test, and antimicrobial activity was determined with the plate microdilution method. The POBT showed no significant difference between sealers with and those without NPs in cervical and apical thirds (p > 0.05). In the middle third, the adhesion force was significant for Endosequence BC Sealer® (p < 0.05). The results showed that the Sa and Sy parameters, when AgNPs were added, did not show a statistically significant difference compared to the groups without nanoparticles (p > 0.05). All tested sealers showed bacterial growth inhibition, but no significant difference was found. Their efficacy, in descending order of antibacterial activity when AgNPs were added, is as follows: EndoSequence® BC SealerTM > Bio-C® Sealer > CeraSeal®. The incorporation of AgNPs into bioceramics improves antimicrobial activity without affecting mechanical properties.
Assuntos
Enterococcus faecalis , Nanopartículas Metálicas , Materiais Restauradores do Canal Radicular , Prata , Propriedades de Superfície , Prata/química , Prata/farmacologia , Nanopartículas Metálicas/química , Materiais Restauradores do Canal Radicular/química , Materiais Restauradores do Canal Radicular/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/crescimento & desenvolvimento , Anti-Infecciosos/farmacologia , Anti-Infecciosos/química , Antibacterianos/farmacologia , Antibacterianos/química , Teste de Materiais , Humanos , Testes de Sensibilidade Microbiana , Cerâmica/química , Cerâmica/farmacologia , Microscopia de Força Atômica , Fosfatos de Cálcio , Combinação de Medicamentos , Óxidos , SilicatosRESUMO
Doping of brushite cements with metal ions can entail many positive effects on biological and physicochemical properties. Cu2+ ions are known to exhibit antibacterial properties and can additionally have different positive effects on cells as trace elements, whereas high Cu2+ concentrations are cytotoxic. For therapeutical applications of bone cement, a combination of good biocompatibility and sufficient mechanical properties is required. Therefore, the aim of this study was to investigate different physicochemical and biological aspects, relevant for application, of a brushite cement with Cu2+-doped ß-tricalcium phosphate, monocalcium phosphate monohydrate and phytic acid as setting retarder. Additionally, the ion release was compared with a cement with citric acid as setting retarder. The investigated cements showed good injectability coefficients, as well as compressive strength values sufficient for application. Furthermore, no antibacterial effects were detected irrespective of the Cu2+ concentration or the bacterial strain. The cell experiments with eluate samples showed that the viability of MC3T3-E1 cells tended to decrease with increasing Cu2+ concentration in the cement. It is suggested that these biological responses are caused by the difference in the Cu2+ release from the hardened cement depending on the solvent medium. Furthermore, the cements showed a steady release of Cu2+ ions to a lesser extent in comparison with a cement with citric acid as setting retarder, where a burst release of Cu2+ was observed. In conclusion, despite the anticipated antibacterial effect of Cu2+-doped cements was lacking and mammalian cell viability was slightly affected, Cu2+-concentrations maintained the physicochemical properties as well as the compressive strength of cements and the slow ion release from cements produced with phytic acid is considered advantageous compared to citric acid-based formulations.
Assuntos
Cimentos Ósseos , Fosfatos de Cálcio , Cobre , Teste de Materiais , Camundongos , Animais , Cobre/química , Cimentos Ósseos/química , Cimentos Ósseos/farmacologia , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Antibacterianos/química , Antibacterianos/farmacologia , Força Compressiva , Sobrevivência Celular/efeitos dos fármacos , Linhagem Celular , Células 3T3 , Ácido Cítrico/químicaRESUMO
The rising prevalence of bone injuries has increased the demand for minimally invasive treatments. Microbead hydrogels, renowned for cell encapsulation, provide a versatile substrate for bone tissue regeneration. They deliver bioactive agents, support cell growth, and promote osteogenesis, aiding bone repair and regeneration. In this study, we synthesized superparamagnetic iron oxide nanoparticles (Sp) coated with a calcium phosphate layer (m-Sp), achieving a distinctive flower-like micro-cluster morphology. Subsequently, sodium alginate (SA) microbead hydrogels containing m-Sp (McSa@m-Sp) were fabricated using a dropping gelation strategy. McSa@m-Sp is magnetically targetable, enhance cross-linking, control degradation rates, and provide strong antibacterial activity. Encapsulation studies with MC3T3-E1 cells revealed enhanced viability and proliferation. These studies also indicated significantly elevated alkaline phosphatase (ALP) activity and mineralization in MC3T3-E1 cells, as confirmed by Alizarin Red S (ARS) and Von Kossa staining, along with increased collagen production within the McSa@m-Sp microbead hydrogels. Immunocytochemistry (ICC) and gene expression studies supported the osteoinductive potential of McSa@m-Sp, showing increased expression of osteogenic markers including RUNX-2, collagen-I, osteopontin, and osteocalcin. Thus, McSa@m-Sp microbead hydrogels offer a promising strategy for multifunctional scaffolds in bone tissue engineering.
Assuntos
Alginatos , Regeneração Óssea , Fosfatos de Cálcio , Proliferação de Células , Hidrogéis , Osteogênese , Alginatos/química , Alginatos/farmacologia , Animais , Camundongos , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Osteogênese/efeitos dos fármacos , Hidrogéis/química , Hidrogéis/farmacologia , Regeneração Óssea/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Engenharia Tecidual/métodos , Linhagem Celular , Nanopartículas Magnéticas de Óxido de Ferro/química , Antibacterianos/farmacologia , Antibacterianos/químicaRESUMO
BACKGROUND: Alveolar Bone loss occurs frequently during the first six months after tooth extraction. Various studies have proposed different methods to reduce as much as possible the atrophy of the alveolar ridge after tooth extraction. Filling the socket with biomaterials after extraction can reduce the resorption of the alveolar ridge. We compared the height of the alveolar process at the mesial and distal aspects of the extraction site and the resorption rate was calculated after the application of HA/ß-TCP or synthetic co-polymer polyglycolic - polylactic acid PLGA mixed with blood to prevent socket resorption immediately and after tooth extraction. METHODS: The study was conducted on 24 extraction sockets of impacted mandibular third molars bilaterally, vertically, and completely covered, with a thin bony layer. HA/ß-TCP was inserted into 12 of the dental sockets immediately after extraction, and the synthetic polymer PLGA was inserted into 12 of the dental sockets. All sockets were covered completely with a full-thickness envelope flap. Follow-up was performed for one year after extraction, using radiographs and stents for the vertical alveolar ridge measurements. RESULTS: The mean resorption rate in the HA/ß-TCP and PLGA groups was ± 1.23 mm and ± 0.1 mm, respectively. A minimal alveolar bone height reduction of HA/ß-TCP was observed after 9 months, the reduction showed a slight decrease to 0.93 mm, while this rate was 0.04 mm after 9 months in the PLGA group. Moreover, the bone height was maintained after three months, indicating a good HA/ß-TCP graft performance in preserving alveolar bone (1.04 mm) while this rate was (0.04 mm) for PLGA. CONCLUSION: The PLGA graft demonstrated adequate safety and efficacy in dental socket preservation following tooth extraction. However, HA/ß-TCP causes greater resorption at augmented sites than PLGA, which clinicians should consider during treatment planning.
Assuntos
Perda do Osso Alveolar , Substitutos Ósseos , Ácido Láctico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Extração Dentária , Alvéolo Dental , Humanos , Alvéolo Dental/cirurgia , Perda do Osso Alveolar/prevenção & controle , Substitutos Ósseos/uso terapêutico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/uso terapêutico , Masculino , Feminino , Ácido Láctico/uso terapêutico , Adulto , Ácido Poliglicólico/uso terapêutico , Processo Alveolar/patologia , Dente Serotino/cirurgia , Dente Impactado/cirurgia , Seguimentos , Adulto Jovem , Retalhos Cirúrgicos , Materiais Biocompatíveis/uso terapêutico , Aumento do Rebordo Alveolar/métodos , Hidroxiapatitas/uso terapêutico , Mandíbula/cirurgia , Fosfatos de Cálcio/uso terapêutico , Resultado do TratamentoRESUMO
The interconnected structures in a 3D scaffold allows the movement of cells and nutrients. Therefore, this study aimed to investigate the in-vivo bioactivity of 3D-printed ß-tricalcium phosphate (ß-TCP) and hydroxyapatite (HAP) scaffolds that replicate biological bone. This study included 24-week-old male New Zealand white rabbits. A cylindrical bone defect with a diameter of 4.5 mm and a depth of 8 mm was created in the lateral aspect of the distal femur. A 3D-printed scaffold was implanted in the right femur (experimental side), whereas the left femur was kept free of implantation (control side). Micro-CT analysis and histological observations of the bone defect site were conducted at 4, 8, and 12 weeks postoperatively to track the bone repair progress. No evidence of new bone tissue formation was found in the medullary cavity of the bone defect on the control side. In contrast, on the experimental side, the 3D scaffold demonstrated sufficient bioactivity, leading to the growth of new bone tissue. Over time, new bone tissue gradually extended from the periphery toward the center, a phenomenon evident in both micro-CT images and biopsy staining. In the current study, we observed that the cells involved in bone metabolism adhered, spread, and proliferated on our newly designed 3D-printed scaffold with a bone microstructure. Therefore, it is suggested that this scaffold has sufficient bioactivity to induce new bone formation and could be expected to be a more useful artificial bone than the existing version.
Assuntos
Regeneração Óssea , Fosfatos de Cálcio , Fêmur , Impressão Tridimensional , Engenharia Tecidual , Alicerces Teciduais , Microtomografia por Raio-X , Coelhos , Animais , Fosfatos de Cálcio/química , Alicerces Teciduais/química , Masculino , Regeneração Óssea/efeitos dos fármacos , Fêmur/patologia , Engenharia Tecidual/métodos , Osteogênese/fisiologia , Osteogênese/efeitos dos fármacos , Durapatita/química , Osso e Ossos/patologia , Substitutos Ósseos/química , Substitutos Ósseos/farmacologia , Teste de Materiais , Materiais Biocompatíveis/químicaRESUMO
Complex bone diseases such as osteomyelitis, osteosarcoma, and osteoporosis often cause critical-size bone defects that the body cannot self-repair and require an advanced bone graft material to repair. We have fabricated 3D-printed tricalcium phosphate bone scaffolds functionalized with garlic extract (GE). GE was encapsulated in a nanoemulsion (GE-NE) to enhance bioavailability and stability. GE-NE showed â¼73% drug encapsulation efficiency, with an average particle size of 158 nm and a zeta potential of -14.2 mV. Release of GE-NEs from the scaffold displayed a controlled and biphasic release profile at both acidic and physiological mediums. Results from the osteosarcoma study show that GE-NE demonstrated â¼88% reduction in cancer cell growth while exhibiting no cytotoxicity toward bone-forming cells. Interaction for the functionalized scaffold with Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa showed a substantial reduction in bacteria growth by more than 90% compared to the unfunctionalized scaffold. These findings demonstrate the potential of GE-NEs-treated porous scaffolds to treat bone-related diseases, particularly for non-load bearing applications.
Assuntos
Emulsões , Alho , Extratos Vegetais , Impressão Tridimensional , Staphylococcus aureus , Alicerces Teciduais , Alicerces Teciduais/química , Alho/química , Humanos , Staphylococcus aureus/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Emulsões/química , Pseudomonas aeruginosa/efeitos dos fármacos , Linhagem Celular Tumoral , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Antibacterianos/farmacologia , Antibacterianos/química , Tamanho da PartículaRESUMO
Decellularized extracellular matrix (dECM) hydrogels are engineered constructs that are widely-used in the field of regenerative medicine. However, the development of ECM-based hydrogels for bone tissue engineering requires enhancement in its osteogenic properties. For this purpose, we initially employed bone-derived dECM hydrogel (dECM-Hy) in combination with calcium phosphate cement (CPC) paste to improve the biological and structural properties of the dECM hydrogel. A decellularization protocol for bovine bone was developed to prepare dECM-Hy, and the mechanically-tuned dECM/CPC-Hy was built based on both rheological and mechanical characteristics. The dECM/CPC-Hy displayed a double swelling ratio and compressive strength. An interconnected structure with distinct hydroxyapatite crystals was evident in dECM/CPC-Hy. The expression levels of Alp, Runx2 and Ocn genes were upregulated in dECM/CPC-Hy compared to the dECM-Hy. A 14-day follow-up of the rats receiving subcutaneous implanted dECM-Hy, dECM/CPC-Hy and mesenchymal stem cells (MSCs)-embedded (dECM/CPC/MSCs-Hy) showed no toxicity, inflammatory factor expression or pathological changes. Radiography and computed tomography (CT) of the calvarial defects revealed new bone formation and elevated number of osteoblasts-osteocytes and osteons in dECM/CPC-Hy and dECM/CPC/MSCs-Hy compared to the control groups. These findings indicate that the dECM/CPC-Hy has substantial potential for bone tissue engineering.
Assuntos
Cimentos Ósseos , Regeneração Óssea , Fosfatos de Cálcio , Células-Tronco Mesenquimais , Animais , Fosfatos de Cálcio/química , Regeneração Óssea/efeitos dos fármacos , Bovinos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Ratos , Cimentos Ósseos/química , Cimentos Ósseos/farmacologia , Matriz Extracelular Descelularizada/química , Matriz Extracelular Descelularizada/farmacologia , Hidrogéis/química , Hidrogéis/farmacologia , Osteogênese/efeitos dos fármacos , Ratos Sprague-Dawley , Engenharia Tecidual , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Matriz Extracelular/química , Matriz Extracelular/metabolismoRESUMO
ARTICLE TITLE AND BIBLIOGRAPHIC INFORMATION: Effectiveness of Calcium Phosphate derivative agents on the prevention and remineralization of caries among children- A systematic review & meta-analysis of randomized controlled trials. Singal K, Sharda S, Gupta A, Malik VS, Singh M, Chauhan A, Agarwal A, Pradhan P, Singh M. J Evid Based Dent Pract. 2022; 22(3):101746. SOURCE OF FUNDING: Indian Council of Medical Research. TYPE OF STUDY/DESIGN: Systematic review with meta-analysis.
Assuntos
Fosfatos de Cálcio , Cárie Dentária , Fluoretos , Remineralização Dentária , Criança , Humanos , Fosfatos de Cálcio/uso terapêutico , Cariostáticos/uso terapêutico , Cariostáticos/farmacologia , Cariostáticos/administração & dosagem , Cárie Dentária/prevenção & controle , Fluoretos/administração & dosagem , Fluoretos/uso terapêutico , Ensaios Clínicos Controlados Aleatórios como Assunto , Metanálise como Assunto , Revisões Sistemáticas como AssuntoRESUMO
Bacteria can solubilize phosphorus (P) through the secretion of low-molecular-weight organic acids and acidification. However, the genes involved in the production of these organic acids are poorly understood. The objectives of this study were to verify the calcium phosphate solubilization and the production of low-molecular-weight organic acids by diverse genera of phosphate solubilizing bacterial strains (PSBS); to identify the genes related to the synthesis of the organic acids in the genomes of these strains and; to evaluate growth and nutrient accumulation of maize plants inoculated with PSBS and fertilized with Bayóvar rock phosphate. Genomic DNA was extracted for strain identification and annotation of genes related to the organic acids production. A greenhouse experiment was performed with five strains plus 150 mg dm- 3 P2O5 as Bayóvar rock phosphate (BRP) to assess phosphate solubilization contribution to maize growth and nutrition. Paraburkholderia fungorum UFLA 04-21 and Pseudomonas anuradhapurensis UFPI B5-8A solubilized over 60% of Ca phosphate and produced high amounts of citric/maleic and gluconic acids in vitro, respectively. Eleven organic acids were identified in total, although not all strains produced all acids. Besides, enzymes related to the organic acids production were found in all bacterial genomes. Plants inoculated with strains UFPI B5-6 (Enterobacter bugandensis), UFPI B5-8A, and UFLA 03-10 (Paenibacillus peoriae) accumulated more biomass than the plants fertilized with BRP only. Strains UFLA 03-10 and UFPI B5-8A increased the accumulation of most macronutrients, including P. Collectively, the results show that PSBS can increase maize growth and nutrient accumulation based on Bayóvar rock phosphate fertilization.
Assuntos
Bactérias , Fosfatos , Zea mays , Zea mays/crescimento & desenvolvimento , Zea mays/microbiologia , Zea mays/metabolismo , Fosfatos/metabolismo , Bactérias/genética , Bactérias/metabolismo , Bactérias/classificação , Fosfatos de Cálcio/metabolismo , Microbiologia do Solo , Genoma Bacteriano , Desenvolvimento Vegetal , Solubilidade , Gluconatos/metabolismo , Genômica , Fósforo/metabolismo , FilogeniaRESUMO
BACKGROUND: The effects of sodium-glucose cotransporter 2 inhibitors (SGLT2i) on calcium phosphate homeostasis in patients with type 2 diabetes mellitus (T2DM) and chronic kidney disease (CKD) remain uncertain. METHODS: A retrospective observational cohort study of patients with T2DM at CKD stage G3b-5ND who received SGLT2i as compared to control from 1 January 2015 through 31 December 2021 was recruited. Propensity score assignment at 1:3 ratio by logistic regression was done. All patients were followed for 12 months. Outcomes were changes in phosphate level. RESULTS: We analyzed 1,450 SGLT2i users and 4,350 control subjects. At the 12th month, SGLT2i users had a slower increase in phosphate levels (absolute change: -0.01 ± 0.28 vs + 0.14 ± 0.34 mmol/L; percentage change: -0.74 % ± 25.56 vs + 10.88 ± 28.15 %, P for both < 0.001). The proportion of patients with high phosphate was lower with SGLT2i (8.2 % vs 24.6 % increase). In the generalized estimating equation, SGLT2i was linked to a longitudinal reduction in phosphate (B -0.039, P<0.001). CONCLUSIONS: SGLT2i can effectively slow down the progression of phosphate retention in advanced CKD with T2DM.
Assuntos
Fosfatos de Cálcio , Diabetes Mellitus Tipo 2 , Homeostase , Insuficiência Renal Crônica , Inibidores do Transportador 2 de Sódio-Glicose , Humanos , Inibidores do Transportador 2 de Sódio-Glicose/uso terapêutico , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Masculino , Feminino , Insuficiência Renal Crônica/tratamento farmacológico , Insuficiência Renal Crônica/metabolismo , Pessoa de Meia-Idade , Estudos Retrospectivos , Idoso , Homeostase/efeitos dos fármacos , Fosfatos de Cálcio/metabolismoRESUMO
This work investigated the range of substitution of two biologically relevant ions, namely Mn2+ and Co2+, into the structure of ß-tricalcium phosphate, as well as their influence on bone cells response. To this aim, ß-TCP was synthesized by solid state reaction in the presence of increasing amount of the substituent ions. The results of the X-ray diffraction analysis reveal that just limited amounts of these ions can enter into the ß-TCP structure: 15â¯at% and 20â¯at% for cobalt and manganese, respectively. Substitution provokes aggregation of the micrometric particles and reduction of the lattice constants. In particular, the dimension of the c-parameter exhibits a discontinuity at about 10â¯at% for both cations, although with different trend. Moreover, Rietveld refinement demonstrates a clear preference of both manganese and cobalt for the octahedral site (V). The influence of these ions on cell response was tested on osteoblast, osteoclast and endothelial cells. The results indicate that the presence of manganese promotes a good osteoblast viability, significantly enhances the expression of osteoblast key genes and the angiogenic process of endothelial cells, while inhibiting osteoclast resorption. At variance, osteoblast viability appears reduced in the presence of Co samples, on which osteoblast genes reach higher expression than on ß-TCP just in a few cases. On the other hand, the results clearly show that cobalt significantly stimulates the angiogenic process and inhibits osteoclast resorption.
Assuntos
Fosfatos de Cálcio , Cobalto , Manganês , Osteoblastos , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Manganês/química , Manganês/farmacologia , Cobalto/química , Cobalto/farmacologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/citologia , Osteoblastos/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Humanos , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Osteoclastos/citologia , Difração de Raios X , Animais , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , CamundongosRESUMO
Bone regeneration often fails due to implants/grafts lacking vascular supply, causing necrotic tissue and poor integration. Microsurgical techniques are used to overcome this issue, allowing the graft to anastomose. These techniques have limitations, including severe patient morbidity and current research focuses on stimulating angiogenesis in situ using growth factors, presenting limitations, such as a lack of control and increased costs. Non-biological stimuli are necessary to promote angiogenesis for successful bone constructs. Recent studies have reported that bioactive glass dissolution products, such as calcium-releasing nanoparticles, stimulate hMSCs to promote angiogenesis and new vasculature. Moreover, the effect of 3D microporosity has also been reported to be important for vascularisation in vivo. Therefore, we used room-temperature extrusion 3D printing with polylactic acid (PLA) and calcium phosphate (CaP) based glass scaffolds, focusing on geometry and solvent displacement for scaffold recovery. Combining both methods enabled reproducible control of 3D structure, porosity, and surface topography. Scaffolds maintained calcium ion release at physiological levels and supported human mesenchymal stem cell proliferation. Scaffolds stimulated the secretion of vascular endothelial growth factor (VEGF) after 3 days of culture. Subcutaneous implantation in vivo indicated good scaffold integration and blood vessel infiltration as early as one week after. PLA-CaP scaffolds showed increased vessel maturation 4 weeks after implantation without vascular regression. Results show PLA/CaP-based glass scaffolds, made via controlled 3D printing, support angiogenesis and vessel maturation, promising improved vascularization for bone regeneration.
Assuntos
Fosfatos de Cálcio , Vidro , Neovascularização Fisiológica , Poliésteres , Impressão Tridimensional , Alicerces Teciduais , Humanos , Poliésteres/química , Poliésteres/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Alicerces Teciduais/química , Vidro/química , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Animais , Regeneração Óssea/efeitos dos fármacos , Regeneração Óssea/fisiologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Engenharia Tecidual/métodos , Osso e Ossos/irrigação sanguínea , Osso e Ossos/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Camundongos , Porosidade , Proliferação de Células/efeitos dos fármacosRESUMO
Tumor acidity-driven nanomotors may offer robust propulsion for tumor-specific penetrating drug delivery. Herein, an acidity-actuated poly(amino acid) calcium phosphate (CaP) hybrid nanomotor (PCaPmotor) was designed, using a mPEG-PAsp-PPhe@THZ531 micelle (Poly@THZ) for CaP mineralization accompanied by αPD-L1 antibody encapsulation. Dissolution of the CaP layer in an acidic tumor environment gave off heat energy to propel the nanomotor to augment the cellular uptake and penetration into deeply seated cancer cells while facilitating αPD-L1 release. THZ531 delivered by the PCaPmotor inhibited CDK12 and its down-streamed phosphorylation of RNAP-II to increase the cancer immunogenicity events such as the DNA damage, cell apoptosis, immunogenic cell death, lysosomal function disturbance, and MHC-I upregulation. THZ531 and αPD-L1 cosupplied by PCaPmotor significantly increased the frequency of DCs maturation and intratumoral infiltration of CTLs, but the two free drugs did not. Consequently, the PCaP@THZ/αPD-L1 nanomotor resulted in synergistic anticancer immunotherapy in mice. This acid-actuated PCaPmotor represented a new paradigm for penetrating drug delivery.
Assuntos
Fosfatos de Cálcio , Sistemas de Liberação de Medicamentos , Imunoterapia , Fosfatos de Cálcio/química , Animais , Camundongos , Humanos , Linhagem Celular Tumoral , Polímeros/química , Micelas , Antineoplásicos/farmacologia , Antineoplásicos/química , Antineoplásicos/administração & dosagem , Concentração de Íons de Hidrogênio , Neoplasias/terapia , Neoplasias/tratamento farmacológico , Antígeno B7-H1 , Nanopartículas/químicaRESUMO
Osteomyelitis is an inflammation of bone tissue usually caused by pyogenic bacteria. The most recurrent clinical approach consists of bone debridement followed by parenteral administration of antibiotics. However, systemic antibiotic treatment has limitations regarding absorption rate and bioavailability over time. The main challenge of osteomyelitis treatment consists of coupling the persistent infection treatment with the regeneration of the bone debrided. In this work, we developed an injectable drug delivery system based on poloxamer 407 hydrogel containing undoped Mg, Zn-doped tricalcium phosphate (ß-TCP), and teicoplanin, a broad-spectrum antibiotic. We evaluated how the addition of teicoplanin and ß-TCP affected the micellization, gelation, particle size, and surface charge of the hydrogel. Later, we studied the hydrogel degradation and drug delivery kinetics. Finally, the bactericidal, biocompatibility, and osteogenic properties were evaluated through in vitro studies and confirmed by in vivo Wistar rat models. Teicoplanin was found to be encapsulated in the corona portions of the hydrogel micelles, yielding a bigger hydrodynamics radius. The encapsulated teicoplanin showed a sustained release over the evaluated period, enough to trigger antibacterial properties against Gram-positive bacteria. Besides, the formulations were biocompatible and showed bone healing ability and osteogenic properties. Finally, in vivo studies confirmed that the proposed locally injected formulations yielded osteomyelitis treatment with superior outcomes than parenteral administration while promoting bone regeneration. In conclusion, the presented formulations are promising drug delivery systems for osteomyelitis treatment and deserve further technological improvements.