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The fluid loss additive is to prevent the cement slurry from filtrating water to the formation under pressure. 2-Acrylamido-2-methylpropanesulfonic acid (AMPS)-based fluid loss additive mainly works by adsorbing on the surface of cement particles. The adsorption affects cement hydration. In this paper, the effect of one kind of AMPS-based fluid loss additive (A-FLA) on the hydration of oil well cement was studied. The water loss, setting time, thickening time, and compressive strength of cement slurry with various amounts of FLA were measured. In addition, the hydration heat of the cement slurry, FLA adsorption isotherm on cement particles, and hydration minerals were studied. The results showed that A-FLA had a good water loss control ability. The water loss of the cement slurry decreased slowly with the increase of A-FLA dosage when the adsorption capacity exceeded 4.47 mg/g. The low adsorption capacity of A-FLA (less than 4.47 mg/g) had a significant impact on the thickening time. With an adsorption capacity greater than 4.47 mg/g, the thickening time varied minimally. A-FLA mainly delayed the hydration of C3S at 1 day and reduced the production amorphous phase.
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Untreated osteochondral defects will develop into osteoarthritis, affecting patients' quality of life. Since articular cartilage and subchondral bone exhibit distinct biological characteristics, repairing osteochondral defects remains a major challenge. Previous studies have tried to fabricate multilayer scaffolds with traditional methods or 3D printing technology. However, the efficacy is unsatisfactory because of poor control over internal structures or a lack of integrity between adjacent layers, severely compromising repair outcomes. Therefore, there is a need for a biomimetic scaffold that can simultaneously boost osteochondral defect regeneration in both structure and function. Herein, an integrated bilayer scaffold with precisely controlled structures is successfully 3D-printed in one step via digital light processing (DLP) technology. The upper layer has both 'lotus- and radial-' distribution pores, and the bottom layer has 'lotus-' pores to guide and facilitate the migration of chondrocytes and bone marrow mesenchymal stem cells, respectively, to the defect area. Tuning pore sizes could modulate the mechanical properties of scaffolds easily. Results show that 3D-printed porous structures allow significantly more cells to infiltrate into the area of 'lotus- and radial-' distribution pores during cell migration assay, subcutaneous implantation, andin situtransplantation, which are essential for osteochondral repair. Transplantation of this 3D-printed bilayer scaffold exhibits a promising osteochondral repair effect in rabbits. Incorporation of Kartogenin into the upper layer of scaffolds further induces better cartilage formation. Combining small molecules/drugs and precisely size-controlled and layer-specific porous structure via DLP technology, this 3D-printed bilayer scaffold is expected to be a potential strategy for osteochondral regeneration.
Assuntos
Cartilagem Articular , Alicerces Teciduais , Humanos , Animais , Coelhos , Alicerces Teciduais/química , Biomimética , Qualidade de Vida , Movimento Celular , Impressão Tridimensional , Engenharia Tecidual/métodosRESUMO
Dental pulp regeneration is ideal for irreversible pulp or periapical lesions, and in situ stem cell therapy is one of the most effective therapies for pulp regeneration. In this study, we provided an atlas of the non-cultured and monolayer cultured dental pulp cells with single-cell RNA sequencing and analysis. Monolayer cultured dental pulp cells cluster more closely together than non-cultured dental pulp cells, suggesting a lower heterogeneous population with relatively consistent clusters and similar cellular composition. We successfully fabricated hDPSC-loaded microspheres by layer-by-layer photocuring with a digital light processing (DLP) printer. These hDPSC-loaded microspheres have improved stemness and higher multi-directional differentiation potential, including angiogenic, neurogenic, and odontogenic differentiation. The hDPSC-loaded microspheres could promote spinal cord regeneration in rat spinal cord injury models. Moreover, in heterotopic implantation tests on nude mice, CD31, MAP2, and DSPP immunofluorescence signals were observed, implying the formation of vascular, neural, and odontogenetic tissues. In situ experiments in minipigs demonstrated highly vascularized dental pulp and uniformly arranged odontoblast-like cells in root canals of incisors. In short, hDPSC-loaded microspheres can promote full-length dental pulp regeneration at the root canals' coronal, middle, and apical sections, particularly for blood vessels and nerve formation, which is a promising therapeutic strategy for necrotic pulp.
Assuntos
Polpa Dentária , Regeneração , Camundongos , Ratos , Suínos , Animais , Porco Miniatura , Microesferas , Camundongos Nus , Células-Tronco , Diferenciação Celular , Medula Espinal , Células CultivadasRESUMO
Ratiometric fluorescent molecularly imprinted polymer (MIP) sensors hold great promise in many bioanalytical areas because of their high sensitivity and selectivity as well as excellent self-referencing and visual detection capability. However, their synthetic strategies are rather limited and the development of such optosensing MIPs that can directly and selectively quantify small organic analytes in complex biological samples remains a formidable challenge owing to the complexity of sample matrices. Herein, a versatile and modular strategy to obtaining well-defined ratiometric fluorescent MIP microspheres capable of directly and selectively detecting an organic herbicide [2,4-dichlorophenoxyacetic acid (2,4-D)] in undiluted pure milks is described. First, it involves the synthesis of uniform "living" polymer particles via RAFT precipitation polymerization, their successive well-controlled grafting of a polymer shell labeled with red CdTe QDs (being inert to 2,4-D) and an MIP shell labeled with green 4-nitrobenzo[c][1,2,5]oxadiazole (NBD) units (showing fluorescence "light-up" upon binding 2,4-D) via surface-initiated RAFT polymerization, and final grafting of hydrophilic poly(N-isopropylacrylamide) brushes via an efficient coupling reaction (i.e., RAFT coupling chemistry). The resulting hydrophilic dual fluorescent MIP particles showed excellent photostability and reusability. They exhibited obvious analyte binding-induced "turn-on"-type ratiometric fluorescence (and color) change and high 2,4-D optosensing selectivity and sensitivity in pure bovine milk (with a detection limit of 0.13 µM). Moreover, they were directly applied to 2,4-D determination in undiluted pure goat milk with good recoveries (96.0-103.2%) and high accuracy (RSD = 1.5-5.5%), even in the presence of several analogues of 2,4-D. The general applicability of our strategy was also demonstrated. This study paves the way for efficiently developing various advanced MIP optosensors (of easily tunable structures and desired properties) highly promising in many bioanalytical applications.
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It is still a challenge for existing bioprinting technologies to fabricate organs suitable for implantation, mainly due to the inability to recapitulate the organs' complex anatomical structures, mechanical properties, and biological functions. Additionally, the failure to create 3D constructs with interconnected microchannels for long-range mass transportation that limits the clinical applications of 3D printing technologies. Here, a new method was developed to print functional living skin (FLS) using a newly designed biomimetic bioink (GelMA/HA-NB/LAP) and digital light processing (DLP)-based 3D printing technology. The FLS possess interconnected microchannels that facilitates cell migration, proliferation and neo-tissue formation. The GelMA/HA-NB/LAP bioink, composed of gelatin methacrylate (GelMA), N-(2-aminoethyl)-4-(4-(hydroxymethyl)-2-methoxy-5-nitrosophenoxy) butanamide (NB) linked hyaluronic acid (HA-NB) and photo-initiator lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP). The bioink demonstrated its rapid gelation kinetics, tunable mechanical properties, good biocompatibility and tissue adhesion. The DLP-based 3D printing technology provides a rapid method to precisely position clusters of human skin fibroblasts (HSFs) and human umbilical vein endothelial cells (HUVECs) with high cell viability to form FLS. The FLS promotes skin regeneration and efficient neovascularization by mimicking the physiological structure of natural skin, and it can also be easily handled and implanted onto the wound site due to its strong mechanical and bio-adhesive properties. Moreover, in vivo study demonstrated that the living skin exhibited instant defense function and had superior performance in promoting dermal regeneration with skin appendages in large animals. This study provides a rapid and mass production method of functional living organs for future clinical applications.
Assuntos
Bioimpressão , Animais , Gelatina , Humanos , Impressão Tridimensional , Regeneração , PeleRESUMO
Current biomaterials and tissue engineering techniques have shown a promising efficacy on full-thickness articular cartilage defect repair in clinical practice. However, due to the difficulty of implanting biomaterials or tissue engineering constructs into a partial-thickness cartilage defect, it remains a challenge to provide a satisfactory cure in joint surface regeneration in the early and middle stages of osteoarthritis. In this study, we focused on a ready-to-use tissue-adhesive joint surface paint (JS-Paint) capable of promoting and enhancing articular surface cartilage regeneration. The JS-Paint is mainly composed of N-(2-aminoethyl)-4-(4-(hydroxymethyl)-2-methoxy-5-nitrosophenoxy) butanamide (NB)-coated silk fibroin microparticles and possess optimal cell adhesion, migration, and proliferation properties. NB-modified silk fibroin microparticles can directly adhere to the cartilage and form a smooth layer on the surface via the photogenerated aldehyde group of NB reacting with the -NH2 groups of the cartilage tissue. JS-Paint treatment showed a significant promotion of cartilage regeneration and restored the smooth joint surface at 6 weeks postsurgery in a rabbit model of a partial-thickness cartilage defect. These findings revealed that silk fibroin can be utilized to bring about a tissue-adhesive paint. Thus, the JS-Paint strategy has some great potential to enhance joint surface regeneration and revolutionize future therapeutics of early and middle stages of osteoarthritis joint ailments.
Assuntos
Cartilagem Articular/fisiologia , Fibroínas/química , Regeneração/efeitos dos fármacos , Adesivos Teciduais/química , Animais , Álcoois Benzílicos/química , Álcoois Benzílicos/efeitos da radiação , Álcoois Benzílicos/toxicidade , Cartilagem Articular/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Fibroínas/toxicidade , Articulações/patologia , Articulações/cirurgia , Coelhos , Adesivos Teciduais/efeitos da radiação , Adesivos Teciduais/toxicidade , Raios UltravioletaRESUMO
Molecularly imprinted polymer (MIP)-based optosensing materials capable of direct, reliable, and highly selective detection of small organic analytes in complex aqueous samples hold great promise in many bioanalytical applications, but their development remains a challenging task. Addressing this issue, well-defined hydrophilic "turn-on"-type ratiometric fluorescent MIP microspheres are developed via a versatile and modular strategy based on the controlled/"living" radical polymerization method. Its general principle was demonstrated by the synthesis of red CdTe quantum dot (QD)-labeled silica particles with surface-bound atom transfer radical polymerization (ATRP)-initiating groups via the one-pot sol-gel reaction and their successive grafting of a thin fluorescent 2,4-D (an organic herbicide)-MIP layer (labeled with green organic fluorophores bearing both nitrobenzoxadiazole (NBD) and urea interacting groups) and hydrophilic poly(glyceryl monomethacrylate) (PGMMA) brushes via surface-initiated ATRP. The introduction of PGMMA brushes and rationally selected dual fluorescence labeling (i.e., red CdTe QDs being inert to 2,4-D and green NBD showing fluorescence "light-up" upon binding 2,4-D) onto MIP particles afforded them excellent complex aqueous sample-compatibility (due to their largely enhanced hydrophilicity) and analyte binding-induced "turn-on"-type ratiometric fluorescence changes, respectively. Such advanced MIP particles proved to be promising optosensing materials, which had a detection limit of 0.13 µM and showed obvious fluorescent color change upon binding different concentrations of 2,4-D in the undiluted pure milk. Moreover, they were successfully applied for direct and highly selective quantification of 2,4-D in the undiluted pure goat and bovine milks with good recoveries (97.9%-104.5%), even in the presence of several analogues of 2,4-D.
Assuntos
Ácido 2,4-Diclorofenoxiacético/análise , Contaminação de Alimentos/análise , Herbicidas/análise , Leite/química , Animais , Compostos de Cádmio/química , Bovinos , Fluorescência , Cabras , Interações Hidrofóbicas e Hidrofílicas , Metilmetacrilatos/química , Microesferas , Impressão Molecular , Pontos Quânticos/química , Dióxido de Silício/química , Telúrio/químicaRESUMO
Changes in the stiffness of chondrocyte extracellular matrix (ECM) are involved in the pathological progression of osteoarthritis (OA). However, the downstream responses of cartilage ECM stiffness are still unclear. YAP (Yes-associated protein) has been extensively studied as a mechanotransducer, we thus hypothesized that by targeting the downstream molecule activity of ECM stiffness could maintain chondrocyte phenotype and prevent cartilage degeneration in OA. Here, we showed that human cartilage matrix stiffened during pathological progression of OA, and the chondrocyte YAP activity was associated with ECM stiffness. We then mimicked the physiological and pathological stiffness of human cartilage by using PDMS-based substrates, and found that YAP was activated in chondrocytes seeded on stiff substrate, gradually losing their phenotype. In addition, it was observed that YAP was also significantly activated in mice OA development, and conditional knockout (cKO) of YAP in mice preserved collagen II expression and protected cartilage from degeneration in the OA model. Furthermore, intra-articular injection of YAP-selective inhibitor, Verteporfin, significantly maintained cartilage homeostasis in mice OA model. This study indicates that the application of mechanotransducer-targeted drugs could be a potential therapeutic approach for cartilage repair in OA.
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Cartilagem Articular , Quitosana , Osteoartrite , Animais , Condrócitos , Camundongos , Microesferas , Osteoartrite/tratamento farmacológico , VerteporfinaRESUMO
Characterized by their slow adhering property, skeletal muscle myogenic progenitor cells (MPCs) have been widely utilized in skeletal muscle tissue engineering for muscle regeneration, but with limited efficacy. Skeletal muscle regeneration is regulated by various cell types, including a large number of rapidly adhering cells (RACs) where their functions and mechanisms are still unclear. In this study, we explored the function of RACs by co-culturing them with MPCs in a biomimetic skeletal muscle organoid system. Results showed that RACs promoted the myogenic potential of MPCs in the organoid. Single-cell RNA-Seq was also performed, classifying RACs into 7 cell subtypes, including one newly described cell subtype: teno-muscular cells (TMCs). Connectivity map of RACs and MPCs subpopulations revealed potential growth factors (VEGFA and HBEGF) and extracellular matrix (ECM) proteins involvement in the promotion of myogenesis of MPCs during muscle organoid formation. Finally, trans-well experiments and small molecular inhibitors blocking experiments confirmed the role of RACs in the promotion of myogenic differentiation of MPCs. The RACs reported here revealed complex cell diversity and connectivity with MPCs in the biomimetic skeletal muscle organoid system, which not only offers an attractive alternative for disease modeling and in vitro drug screening but also provides clues for in vivo muscle regeneration.
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Desenvolvimento Muscular/genética , Músculo Esquelético/metabolismo , Mioblastos/metabolismo , Organoides/citologia , Animais , Diferenciação Celular/genética , Proliferação de Células/genética , Análise por Conglomerados , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Fator de Crescimento Semelhante a EGF de Ligação à Heparina/genética , Fator de Crescimento Semelhante a EGF de Ligação à Heparina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/citologia , Mioblastos/citologia , Organoides/ultraestrutura , RNA-Seq , Análise de Célula Única , Transcriptoma/genética , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Uncontrollable bleeding is a major problem in surgical procedures and after major trauma. Existing hemostatic agents poorly control hemorrhaging from traumatic arterial and cardiac wounds because of their weak adhesion to wet and mobile tissues. Here we design a photo-reactive adhesive that mimics the extracellular matrix (ECM) composition. This biomacromolecule-based matrix hydrogel can undergo rapid gelling and fixation to adhere and seal bleeding arteries and cardiac walls after UV light irradiation. These repairs can withstand up to 290 mm Hg blood pressure, significantly higher than blood pressures in most clinical settings (systolic BP 60-160 mm Hg). Most importantly, the hydrogel can stop high-pressure bleeding from pig carotid arteries with 4~ 5 mm-long incision wounds and from pig hearts with 6 mm diameter cardiac penetration holes. Treated pigs survived after hemostatic treatments with this hydrogel, which is well-tolerated and appears to offer significant clinical advantage as a traumatic wound sealant.