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1.
ACS Appl Mater Interfaces ; 16(35): 45956-45968, 2024 Sep 04.
Article in English | MEDLINE | ID: mdl-39182190

ABSTRACT

Osteoporotic bone defects are difficult to repair in elderly patients. This study aimed to repair osteoporotic bone defects using a combination of bone tissue engineering (BTE) and drug delivery systems (DDS). Herein, honeycomb granules (HCGs) composed of carbonate apatite microspheres were fabricated as BTE scaffolds. Each HCG possesses hexagonal macropores and abundant interconnected micropores between the microspheres. Owing to these multiscale interconnected pores, HCGs can readily contain antibodies against sclerostin (Scl), which causes imbalances in bone homeostasis. Anti-Scl antibody-loaded HCGs (Scl-Ab-HCGs) regulate the release of Scl-Abs in response to the pH of the osteoporotic environment. In ovariectomized rabbit osteoporotic femurs, HCG monotherapy forms new bone with less osteocyte damage (fewer empty bone lacunae) and fewer osteoclasts than osteoporotic bone; however, it is insufficient to prevent receptor activator of nuclear factor-kappa B ligand (RANKL) overexpression. Consequently, HCG monotherapy restores bone quantity better than no treatment but not to normal levels. In contrast, new bone tissue formed by Scl-Ab-HCG-based DDS predominantly expresses osteocalcin rather than RANKL, similar to normal bone, and shows a similar osteocyte apoptosis level, bone quantity, and osteoclast number as normal bone. Thus, Scl-Ab-HCG-based DDS is a promising approach for osteoporotic bone defect repair.


Subject(s)
Apatites , Drug Delivery Systems , Osteoporosis , Tissue Scaffolds , Animals , Rabbits , Apatites/chemistry , Osteoporosis/drug therapy , Osteoporosis/pathology , Tissue Scaffolds/chemistry , Female , Tissue Engineering , Bone Regeneration/drug effects , Humans , Adaptor Proteins, Signal Transducing/metabolism
2.
Int J Nanomedicine ; 19: 7709-7727, 2024.
Article in English | MEDLINE | ID: mdl-39099788

ABSTRACT

Introduction: Dysregulated calcium homeostasis and consequentially aberrant Ca2+ signalling could enhance survival, proliferation and metastasis in various cancers. Despite rapid development in exploring the ion channel functions in relation to cancer, most of the mechanisms accounting for the impact of ion channel modulators have yet to be fully clarified. Although harnessing small interfering RNA (siRNA) to specifically silence gene expression has the potential to be a pivotal approach, its success in therapeutic intervention is dependent on an efficient delivery system. Nanoparticles have the capacity to strongly bind siRNAs. They remain in the circulation and eventually deliver the siRNA payload to the target organ. Afterward, they interact with the cell surface and enter the cell via endocytosis. Finally, they help escape the endo-lysosomal degradation system prior to unload the siRNAs into cytosol. Carbonate apatite (CA) nanocrystals primarily is composed of Ca2+, carbonate and phosphate. CA possesses both anion and cation binding domains to target negatively charged siRNA molecules. Methods: Hybrid CA was synthesized by complexing CA NPs with a hydrophilic polysaccharide - hyaluronic acid (HA). The average diameter of the composite particles was determined using Zetasizer and FE-SEM and their zeta potential values were also measured. Results and Discussion: The stronger binding affinity and cellular uptake of a fluorescent siRNA were observed for HA-CA NPs as compared to plain CA NPs. Hybrid CA was electrostatically bound individually and combined with three different siRNAs to silence expression of calcium ion channel and transporter genes, TRPC6, TRPM8 and SLC41A1 in a human breast cancer cell line (MCF-7) and evaluate their potential for treating breast cancer. Hybrid NPs carrying TRPC6, TRPM8 and SLC41A1 siRNAs could significantly enhance cytotoxicity both in vitro and in vivo. The resultant composite CA influenced biodistribution of the delivered siRNA, facilitating reduced off target distribution and enhanced breast tumor targetability.


Subject(s)
Apatites , Breast Neoplasms , Hyaluronic Acid , Nanoparticles , RNA, Small Interfering , Humans , Apatites/chemistry , Apatites/pharmacology , RNA, Small Interfering/chemistry , RNA, Small Interfering/pharmacokinetics , RNA, Small Interfering/administration & dosage , RNA, Small Interfering/pharmacology , RNA, Small Interfering/genetics , Hyaluronic Acid/chemistry , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Nanoparticles/chemistry , Female , Animals , Cell Survival/drug effects , Cell Line, Tumor , MCF-7 Cells , Cell Proliferation/drug effects , Mice
3.
J Dent ; 149: 105292, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39111537

ABSTRACT

OBJECTIVES: In this study, we used atomic force microscopy (AFM) to quantify the size of surface pore apertures of enamel white spot lesions and then demonstrated the penetration of fluorapatite nanocrystals (nFA) into the subsurface of these lesions. METHODS: For the porosity study, enamel lesions were created on three sound human teeth using a demineralizing gel for 8 days. The interface between sound enamel and the artificial lesion was analyzed by AFM. To visualize the penetration of nFA tagged with a calcium-binding fluorophore (Fluo-4) into the subsurface of white spot lesions, we used two-photon microscopy. Sixteen extracted human teeth with either active, natural, or in vitro-created carious lesions in enamel were randomly divided into three groups. The teeth were treated for 2 min with either a suspension of tagged nFA crystals, Fluo-4 alone, or deionized water, and left for 30 min before being washed with distilled water and examined microscopically. RESULTS: A greater concentration of surface pores with larger areas was observed on the in vitro demineralized enamel (29 % of pores greater than 1.0 µm2) when compared with the adjacent sound enamel (8 % of pores greater than 1.0 µm2) (p=0.012, Fisher exact test). In vitro and natural lesions treated with tagged nFA showed fluorescence at depths ranging from 50 to 170 µm, demonstrating penetration of the nFA into the lesion subsurface. The lesions treated with Fluo-4 alone with no crystals showed mostly surface fluorescence (restricted to the outer 25 µm), while those treated with deionized water showed minimal (restricted to the outer 20 µm) to no fluorescence. CONCLUSION: We have demonstrated the use of AFM to quantify the surface pore apertures and two-photon microscopy to visualize nFA crystals in the subsurface of non-cavitated enamel lesions. CLINICAL SIGNIFICANCE: The restoration of the subsurface of non-cavitated caries lesions is a clinical challenge. This study demonstrated that a 2 min application of nFA could penetrate through the surface apertures of non-cavitated enamel lesions into their subsurface.


Subject(s)
Apatites , Dental Caries , Dental Enamel , Microscopy, Atomic Force , Nanoparticles , Humans , Dental Enamel/drug effects , Dental Enamel/metabolism , Nanoparticles/chemistry , Apatites/chemistry , Porosity , Dental Caries/pathology , Surface Properties , Proof of Concept Study , Tooth Demineralization
4.
Environ Sci Technol ; 58(32): 14541-14554, 2024 Aug 13.
Article in English | MEDLINE | ID: mdl-39058062

ABSTRACT

Soil minerals influence the biogeochemical cycles of fluoride (F) and phosphorus (P), impacting soil quality and bioavailability to plants. However, the cooperative mechanisms of soil minerals in governing F and P in the soil environment remain a grand challenge. Here, we reveal the essential role of a typical soil mineral, montmorillonite (Mt), in the cycling and fate of F and P. The results show that the enrichment of metal sites on the Mt surface promotes the mineralization of F to the fluorapatite (FAP) phase, thereby remaining stable in the environment, simultaneously promoting P release. This differential behavior leads to a reduction in the level of F pollution and an enhancement of P availability. Moreover, solid-state NMR and HRTEM observations confirm the existence of metastable F-Ca-F intermediates, emphasizing the pivotal role of Mt surface sites in regulating crystallization pathways and crystal growth of FAP. Furthermore, the in situ atomic force microscopy and theoretical calculations reveal molecular fractionation mechanisms and adsorption processes. It is observed that a competitive relationship exists between F and P at the Mt interface, highlighting the thermodynamically advantageous pathway of forming metastable intermediates, thereby governing the activity of F and P in the soil environment at a molecular level. This work paves the way to reveal the important role of clay minerals as a mineralization matrix for soil quality management and offers new strategies for modulating F and P dynamics in soil ecosystems.


Subject(s)
Fluorides , Phosphorus , Fluorides/chemistry , Phosphorus/chemistry , Soil/chemistry , Clay/chemistry , Apatites/chemistry , Bentonite/chemistry , Adsorption , Minerals/chemistry , Aluminum Silicates/chemistry
5.
Langmuir ; 40(31): 16557-16570, 2024 Aug 06.
Article in English | MEDLINE | ID: mdl-39056438

ABSTRACT

Nanocrystalline apatites have been intensively studied for decades, not only for their well-known mimesis of bone apatite but also for applicative purposes, whether as biomaterials for skeletal repair or more recently for a variety of nanomedical applications enabled by their peculiar surface characteristics. Particularly, ion-doped apatites are of great interest because the incorporation of foreign ions in the composition of apatite (nano)crystals alters the bulk and surface properties, modifying their ability to interact with the external environment. This is clearly seen in the physiology of bone tissue, whose mineral phase, a low crystallinity apatitic phase, can dynamically exchange ions with cells, thus driving bone metabolism. Taking bone mineral as a model, the present work describes the development of Mg-doped hydroxyapatite nanoparticles, exploiting hydrothermal synthesis to achieve extents of Mg2+ doping hardly achieved before and using citrate to develop stable apatite colloidal dispersions. Morphological and physicochemical analyses, associated with in-depth investigation of ions populating the apatitic lattice and the nonapatitic surface layer, concurred to demonstrate the cooperative presence of Mg2+ and citrate ions, affecting the dynamic ion retention/release mechanisms. Achieving high Mg2+ doping rates and understanding how Mg doping translates into surface activation of apatite-based nanoparticles is expected to foster the design of novel smart and tunable devices, to adsorb and release ionic species and cargo molecules, with potential innovations in the biomedical field or even beyond, as in catalysis or for environmental remediation.


Subject(s)
Magnesium , Nanoparticles , Nanoparticles/chemistry , Magnesium/chemistry , Apatites/chemistry , Durapatite/chemistry , Surface Properties , Bone and Bones/chemistry , Particle Size
6.
Braz Dent J ; 35: 5858, 2024.
Article in English | MEDLINE | ID: mdl-39045991

ABSTRACT

ER:YAG laser and experimental resin-based dental adhesive loaded with functionalized carbonated apatite filler were used in this study to evaluate the dentin interaction in terms of penetration and occlusion of the dentinal tubules aiding in the control of dentin hypersensitivity (DH). Spheroidal Carbonated apatite nanoparticles (N-CAP), with an average size of 20±5 nm diameter, were synthesized, characterized, and incorporated in a universal adhesive "All Bond Universal, Bisco, USA", in (2% weight) concentration. Er:YAG laser "Lightwalker, FOTONA, EU" was adjusted to an energy output of 40mJ/ pulse and pulse repetition of 10 Hz for 10 seconds. Dentin specimens were prepared from the buccal surface of 75 extracted sound human molars. The specimens were randomly divided into five groups (n=15) according to the surface treatment: Group (L): Laser only; Group (LB): Laser in combination with adhesive; Group (LBN): Laser in combination with adhesive loaded with N-CAP; Group (B): adhesive only; and Group (BN): adhesive loaded with N-CAP. Depth of penetration and occlusion of the dentinal tubules were assessed using Environmental Scanning Electron Microscope Examination (ESEM). One-way ANOVA was used to compare groups, followed by a pairwise test for multiple comparisons (α=0.05). Groups (LB), and (LBN) showed the highest mean of dentinal tubules' penetration, with a non-significant difference between them. In contrast, the specimens treated with laser only (L) showed the most minor penetration. The employment of ER-YAG laser irradiation with the adhesive loaded with N-CAP was evaluated to be effective in penetrating and occluding the opened dentinal tubules.


Subject(s)
Lasers, Solid-State , Lasers, Solid-State/therapeutic use , Humans , Apatites/chemistry , Dentin , Microscopy, Electron, Scanning , Nanoparticles/chemistry , Dentin Sensitivity , In Vitro Techniques , Dental Cements/chemistry
7.
Nat Commun ; 15(1): 5273, 2024 Jun 20.
Article in English | MEDLINE | ID: mdl-38902270

ABSTRACT

Vertebrates use the phosphate mineral apatite in their skeletons, which allowed them to develop tissues such as enamel, characterized by an outstanding combination of hardness and elasticity. It has been hypothesized that the evolution of the earliest vertebrate skeletal tissues, found in the teeth of the extinct group of conodonts, was driven by adaptation to dental function. We test this hypothesis quantitatively and demonstrate that the crystallographic order increased throughout the early evolution of conodont teeth in parallel with morphological adaptation to food processing. With the c-axes of apatite crystals oriented perpendicular to the functional feeding surfaces, the strongest resistance to uniaxial compressional stress is conferred along the long axes of denticles. Our results support increasing control over biomineralization in the first skeletonized vertebrates and allow us to test models of functional morphology and material properties across conodont dental diversity.


Subject(s)
Biological Evolution , Biomineralization , Fossils , Tooth , Biomineralization/physiology , Animals , Tooth/metabolism , Tooth/chemistry , Apatites/metabolism , Apatites/chemistry , Dental Enamel/chemistry , Dental Enamel/metabolism , Vertebrates
8.
J Oral Biosci ; 66(2): 281-287, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38723946

ABSTRACT

BACKGROUND: The osseointegration of zirconia implants has been evaluated based on their implant fixture bonding with the alveolar bone at the optical microscopic level. Achieving nano-level bonding between zirconia and bone apatite is crucial for superior osseointegration; however, only a few studies have investigated nanoscale bonding. This review outlines zirconia osseointegration, including surface modification, and presents an evaluation of nanoscale zirconia-apatite bonding and its structure. HIGHLIGHT: Assuming osseointegration, the cells produced calcium salts on a ceria-stabilized zirconia substrate. We analyzed the interface between calcium salts and zirconia substrates using transmission electron microscopy and found that 1) the cell-induced calcium salts were bone-like apatite and 2) direct nanoscale bonding was observed between the bone-like apatite and zirconia crystals without any special modifications of the zirconia surface. CONCLUSION: Structural affinity exists between bone apatite and zirconia crystals. Apatite formation can be induced by the zirconia surface. Zirconia bonds directly with apatite, indicating superior osseointegration in vivo.


Subject(s)
Durapatite , Osseointegration , Zirconium , Zirconium/chemistry , Osseointegration/drug effects , Durapatite/chemistry , Surface Properties , Humans , Dental Implants , Apatites/chemistry
9.
J Dent ; 146: 105070, 2024 07.
Article in English | MEDLINE | ID: mdl-38740251

ABSTRACT

OBJECTIVES: The objective of this study was to assess whether zinc-doped fluorapatite (ZnFA) could serve as an effective antimicrobial dental bone filler for bone regeneration compared to autografts. METHODS: FA and 2 % zinc-doped FA (2ZnFA) were synthesized and characterized in-house. Compressed and sintered FA and 2ZnFA disks were incubated with bacteria to assess antimicrobial properties. Adipose-derived stem cells were cultured on these discs to evaluate the surfaces' ability to support cell growth and promote osteogenic differentiation. Surfaces exhibiting the highest expressions of the bone markers osteopontin and osteocalcin were selected for an in vivo study in a rat mandibular defect model. Twenty rats were divided into 5 groups, equally, and a 5 mm surgical defect of the jaw was left untreated or filled with 2ZnFA, FA, autograft, or demineralized bone matrix (DBM). At 12 weeks, the defects and surrounding tissues were harvested and subjected to microCT and histological evaluations. RESULTS: Standard techniques such as FTIR, ICP-MS, fluoride probe, and XRD revealed the sintered FA and ZnFA's chemical compositions and structures. Bacterial studies revealed no significant differences in surface bacterial adhesion properties between FA and 2ZnFA, but significantly fewer bacterial loads than control titanium discs (p < 0.05). Cell culture data confirmed that both surfaces could support cell growth and promote the osteogenic differentiation of stem cells. MicroCT analysis confirmed statistical similarities in bone regeneration within FA, 2ZnFA, and autograft groups. CONCLUSION: The data suggests that both FA and 2ZnFA could serve as alternatives to autograft materials, which are the current gold standard. Moreover, these bone fillers outperformed DBM, an allograft material commonly used as a dental bone void filler. CLINICAL SIGNIFICANCE: The use of FA or 2ZnFA for treating mandibular defects led to bone regeneration statistically similar to autograft repair and significantly outperformed the widely used dental bone filler, DBM. Additional translational research may confirm FA-based materials as superior substitutes for existing synthetic bone fillers, ultimately enhancing patient outcomes.


Subject(s)
Apatites , Bone Regeneration , Cell Differentiation , Osteogenesis , Tissue Scaffolds , Zinc , Animals , Apatites/chemistry , Apatites/pharmacology , Bone Regeneration/drug effects , Rats , Tissue Scaffolds/chemistry , Osteogenesis/drug effects , Cell Differentiation/drug effects , Bone Substitutes/pharmacology , Osteopontin , Stem Cells/drug effects , Mandible/surgery , Mandible/diagnostic imaging , X-Ray Microtomography , Osteocalcin , Adipose Tissue/cytology , Anti-Infective Agents/pharmacology , Cell Proliferation/drug effects , Male , Cells, Cultured , Bone Transplantation/methods , Autografts , Spectroscopy, Fourier Transform Infrared
10.
Int J Periodontics Restorative Dent ; 44(3): 257-266, 2024 05 24.
Article in English | MEDLINE | ID: mdl-38787711

ABSTRACT

Bone graft materials are often used in implant treatment to optimize functional and esthetic outcomes. The requirements for bone grafting materials are the ability to maintain space for bone regeneration to occur and the capability of being resorbed by osteoclasts and replaced with new bone tissue occurring in passive chemolysis and bone remodeling. Carbonate apatite (CO3Ap) granules (Cytrans Granules, GC) are a chemically synthetic bone graft material similar to autogenous bone minerals and more biocompatible than allografts and xenografts. The aim of this report is to evaluate the efficacy of CO3Ap granules in implant treatments when used alone or in combination with autogenous bone. The clinical findings and the radiographic and histologic assessments in three cases of immediate implant placement and lateral and vertical guided bone regeneration are reported. Despite the short-term follow-ups, histologic findings showed that CO3Ap granules were efficiently resorbed and replaced bone in clinical use. Furthermore, the clinical findings showed that CO3Ap granules maintained their morphology around the implant. This limited short-term case report suggests that this bone substitute is effective. However, further clinical studies and long-term reports of this new biomaterial are needed.


Subject(s)
Apatites , Bone Substitutes , Humans , Apatites/chemistry , Bone Regeneration/drug effects , Bone Substitutes/therapeutic use , Bone Transplantation/methods , Guided Tissue Regeneration, Periodontal/methods , Immediate Dental Implant Loading/methods
11.
Environ Res ; 252(Pt 2): 118873, 2024 Jul 01.
Article in English | MEDLINE | ID: mdl-38604484

ABSTRACT

Chemical crystallization granulation in a fluidized bed offers an environmentally friendly technology with significant promise for fluoride removal. This study investigates the impact of stratified pH control in a crystallization granulation fluidized bed for the removal of fluoride and phosphate on a pilot scale. The results indicate that using dolomite as a seed crystal, employing sodium dihydrogen phosphate (SDP) and calcium chloride as crystallizing agents, and controlling the molar ratio n(F):n(P):n(Ca) = 1:5:10 with an upflow velocity of 7.52 m/h, effectively removes fluoride and phosphate. Stratified pH control-maintaining weakly acidic conditions (pH = 6-7) at the bottom and weakly alkaline conditions (pH = 7-8) at the top-facilitates the induction of fluoroapatite (FAP) and calcium phosphate crystallization. This approach reduces groundwater fluoride levels from 9.5 mg/L to 0.2-0.6 mg/L and phosphate levels to 0.1-0.2 mg/L. Particle size analysis, scanning electron microscopy-energy-dispersive X-ray spectroscopy, and X-ray diffraction physical characterizations reveal significant differences in crystal morphology between the top and bottom layers, with the lower layer primarily generating high-purity FAP crystals. Further analysis shows that dolomite-induced FAP crystallization offers distinct advantages. SDP not only dissolves on the dolomite surface to provide active sites for crystallization but also, under weakly acidic conditions, renders both dolomite and FAP surfaces negatively charged. This allows for the effective adsorption of PO43-, HPO42-, and F- anions onto the crystal surfaces. This study provides supporting data for the removal of fluoride from groundwater through induced FAP crystallization in a chemical crystallization pellet fluidized bed.


Subject(s)
Crystallization , Fluorides , Phosphates , Fluorides/chemistry , Hydrogen-Ion Concentration , Phosphates/chemistry , Water Purification/methods , Water Pollutants, Chemical/chemistry , Water Pollutants, Chemical/analysis , Apatites/chemistry , Calcium Phosphates/chemistry , Microscopy, Electron, Scanning
12.
Sci Rep ; 14(1): 8347, 2024 04 09.
Article in English | MEDLINE | ID: mdl-38594297

ABSTRACT

Phosphatized fish fossils occur in various locations worldwide. Although these fossils have been intensively studied over the past decades they remain a matter of ongoing research. The mechanism of the permineralization reaction itself remains still debated in the community. The mineralization in apatite of a whole fish requires a substantial amount of phosphate which is scarce in seawater, so the origin of the excess is unknown. Previous research has shown that alkaline phosphatase, a ubiquitous enzyme, can increase the phosphate content in vitro in a medium to the degree of saturation concerning apatite. We applied this principle to an experimental setup where fish scales were exposed to commercial bovine alkaline phosphatase. We analyzed the samples with SEM and TEM and found that apatite crystals had formed on the remaining soft tissue. A comparison of these newly formed apatite crystals with fish fossils from the Solnhofen and Santana fossil deposits showed striking similarities. Both are made up of almost identically sized and shaped nano-apatites. This suggests a common formation process: the spontaneous precipitation from an oversaturated solution. The excess activity of alkaline phosphatase could explain that effect. Therefore, our findings could provide insight into the formation of well-preserved fossils.


Subject(s)
Alkaline Phosphatase , Apatites , Animals , Cattle , Apatites/chemistry , Phosphates/metabolism , Fossils
13.
J Biomed Mater Res B Appl Biomater ; 112(3): e35397, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38456309

ABSTRACT

In this study, we have formulated a novel apatite bone cements derived from natural sources (i.e. eggshell and fishbone) with improved qualities that is, porosity, resorbability, biological activity, and so forth. The naturally-derived apatite bone cement (i.e. FBDEAp) was prepared by mixing hydroxyapatite (synthesized from fishbone) and tricalcium phosphate (synthesized from eggshell) as a solid phase with a liquid phase (a dilute acidic blend of cement binding accelerator and biopolymers like gelatin and chitosan) with polysorbate (as liquid porogen) to get a desired bone cement paste. The prepared cement paste sets within the clinically acceptable setting time (≤20 min), easily injectable (>85%) through hands and exhibits physiological pH stability (7.3-7.4). The pure apatite phased bone cement was confirmed by x-ray diffraction and Fourier transform infrared spectroscopy analyses. The FBDEAp bone cement possesses acceptable compressive strength (i.e. 5-7 MPa) within trabecular bone range and is resorbable up to 28% in simulated body fluid solution within 12 weeks of incubation at physiological conditions. The FBDEAp is macroporous in nature (average pore size ~50-400 µm) with interconnected pores verified by SEM and micro-CT analyses. The FBDEAp showed significantly increased MG63 cell viability (>125% after 72 h), cell adhesion, proliferation, and key osteogenic genes expression levels (up to 5-13 folds) compared to the synthetically derived, synthetic and eggshell derived as well as synthetic and fishbone derived bone cements. Thus, we strongly believe that our prepared FBDEAp bone cement can be used as potential trabecular bone substitute in orthopedics.


Subject(s)
Bone Substitutes , Chitosan , Apatites/pharmacology , Apatites/chemistry , Bone Substitutes/chemistry , Bone Cements/pharmacology , Bone Cements/chemistry , Calcium Phosphates/chemistry , Durapatite , Chitosan/pharmacology , Chitosan/chemistry , X-Ray Diffraction , Compressive Strength
14.
Br Dent J ; 236(5): 414, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38459336
15.
Biomater Adv ; 159: 213801, 2024 May.
Article in English | MEDLINE | ID: mdl-38401402

ABSTRACT

Obtaining rapid mineralisation is a challenge in current bone graft materials, which has been attributed to the difficulty of guiding the biological processes towards osteogenesis. Amelogenin, a key protein in enamel formation, inspired the design of two intrinsically disordered peptides (P2 and P6) that enhance in vivo bone formation, but the process is not fully understood. In this study, we have elucidated the mechanism by which these peptides induce improved mineralisation. Our molecular dynamics analysis demonstrated that in an aqueous environment, P2 and P6 fold to interact with the surrounding Ca2+, PO43- and OH- ions, which can lead to apatite nucleation. Although P2 has a less stable backbone, it folds to a stable structure that allows for the nucleation of larger calcium phosphate aggregates than P6. These results were validated experimentally in a concentrated simulated body fluid solution, where the peptide solutions accelerated the mineralisation process compared to the control and yielded mineral structures mimicking the amorphous calcium phosphate crystals that can be found in lamella bone. A pH drop for the peptide groups suggests depletion of calcium and phosphate, a prerequisite for intrinsic osteoinduction, while S/TEM and SEM suggested that the peptide regulated the mineral nucleation into lamella flakes. Evidently, the peptides accelerate and guide mineral formation, elucidating the mechanism for how these peptides can improve the efficacy of P2 or P6 containing devices for bone regeneration. The work also demonstrates how experimental mineralisation study coupled with molecular dynamics is a valid method for understanding and predicting in vivo performance prior to animal trials.


Subject(s)
Bone Regeneration , Osteogenesis , Animals , Apatites/chemistry , Peptides/pharmacology , Bone and Bones
16.
Adv Healthc Mater ; 13(13): e2303217, 2024 05.
Article in English | MEDLINE | ID: mdl-38363057

ABSTRACT

Osteochondral defects are often accompanied by excessive reactive oxygen species (ROS) caused by osteoarthritis or acute surgical inflammation. An inflammatory environment containing excess ROS will not only hinder tissue regeneration but also impact the quality of newly formed tissues. Therefore, there is an urgent need to develop scaffolds with both ROS scavenging and osteochondral repair functions to promote and protect osteochondral tissue regeneration. In this work, by using 3D printing technology, a composite scaffold based on cobalt-incorporated chloroapatite (Co-ClAP) bioceramics, which possesses ROS-scavenging activity and can support cell proliferation, adhesion, and differentiation, is developed. Benefiting from the catalytic activity of Co-ClAP bioceramics, the composite scaffold can protect cells from oxidative damage under ROS-excessive conditions, support their directional differentiation, and simultaneously mediate an anti-inflammatory microenvironment. In addition, it is also confirmed by using rabbit osteochondral defect model that the Co-ClAP/poly(lactic-co-glycolic acid) scaffold can effectively promote the integrated regeneration of cartilage and subchondral bone, exhibiting an ideal repair effect in vivo. This study provides a promising strategy for the treatment of defects with excess ROS and inflammatory microenvironments.


Subject(s)
Bone Regeneration , Ceramics , Cobalt , Printing, Three-Dimensional , Tissue Scaffolds , Animals , Rabbits , Tissue Scaffolds/chemistry , Cobalt/chemistry , Ceramics/chemistry , Ceramics/pharmacology , Bone Regeneration/drug effects , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Reactive Oxygen Species/metabolism , Antioxidants/chemistry , Antioxidants/pharmacology , Tissue Engineering/methods , Cell Proliferation/drug effects , Apatites/chemistry , Cell Differentiation/drug effects , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/metabolism
17.
Int J Mol Sci ; 25(3)2024 Jan 25.
Article in English | MEDLINE | ID: mdl-38338772

ABSTRACT

Coating layers consisting of a crystalline apatite matrix with immobilized basic fibroblast growth factor (bFGF) can release bFGF, thereby enhancing bone regeneration depending on their bFGF content. We hypothesized that the incorporation of fluoride ions into apatite crystals would enable the tailored release of bFGF from the coating layer depending on the layer's fluoride content. In the present study, coating layers consisting of fluoride-incorporated apatite (FAp) crystals with immobilized bFGF were coated on a porous collagen sponge by a precursor-assisted biomimetic process using supersaturated calcium phosphate solutions with various fluoride concentrations. The fluoride content in the coating layer increased with the increasing fluoride concentration of the supersaturated solution. The increased fluoride content in the coating layer reduced its solubility and suppressed the burst release of bFGF from the coated sponge into a physiological salt solution. The bFGF release was caused by the partial dissolution of the coating layer and, thus, accompanied by the fluoride release. The concentrations of released bFGF and fluoride were controlled within the estimated effective ranges in enhancing bone regeneration. These findings provide useful design guidelines for the construction of a mineralized, bFGF-releasing collagen scaffold that would be beneficial for bone tissue engineering, although further in vitro and in vivo studies are warranted.


Subject(s)
Apatites , Fluorides , Apatites/chemistry , Fibroblast Growth Factor 2/pharmacology , Collagen/chemistry , Tissue Engineering
18.
Adv Healthc Mater ; 13(12): e2303245, 2024 05.
Article in English | MEDLINE | ID: mdl-38229572

ABSTRACT

The aging global population is generating an ever-increasing demand for bone regeneration. Various materials, including blocks, granules, and sponges, are developed for bone regeneration. However, blocks require troublesome shaping and exhibit poor bone-defect conformities; granules migrate into the surrounding tissues during and after filling of the defect, causing handling difficulties and complications; and sponges contain polymers that are subject to religious restrictions, lack osteoconductivity, and may cause inflammation and allergies. Herein, carbonate apatite chains that overcome the limitations of conventional materials are presented. Although carbonate apatite granules migrate, causing inflammation and ectopic calcification, the chains remain in the defects without causing any complications. The chains conform to the defect shape and transform into 3D porous structures, resulting in faster bone regeneration than that observed using granules. Thus, these findings indicate that even traditional calcium phosphates materials can be converted to state-of-the-art materials via shape control.


Subject(s)
Apatites , Bone Regeneration , Apatites/chemistry , Animals , Bone Regeneration/drug effects , Bone Substitutes/chemistry , Bone Transplantation/methods , Porosity , Mice , Male , Tissue Scaffolds/chemistry
19.
Bone ; 181: 117024, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38266952

ABSTRACT

Functional adaptation refers to the active modification of bone structure according to the mechanical loads applied daily to maintain its mechanical integrity and adapt to the environment. Functional adaptation relates to bone mass, bone mineral density (BMD), and bone morphology (e.g., trabecular bone architecture). In this study, we discovered for the first time that another form of bone functional adaptation of a cortical bone involves a change in bone quality determined by the preferential orientation of apatite nano-crystallite, a key component of the bone. An in vivo rat ulnar axial loading model was adopted, to which a 3-15 N compressive load was applied, resulting in approximately 440-3200 µÉ› of compression in the bone surface. In the loaded ulnae, the degree of preferential apatite c-axis orientation along the ulnar long axis increased in a dose-dependent manner up to 13 N, whereas the increase in BMD was not dose-dependent. The Young's modulus along the same direction was enhanced as a function of the degree of apatite orientation. This finding indicates that bone has a mechanism that modifies the directionality (anisotropy) of its microstructure, strengthening itself specifically in the loaded direction. BMD, a scalar quantity, does not allow for load-direction-specific strengthening. Functional adaptation through changes in apatite orientation is an excellent strategy for bones to efficiently change their strength in response to external loading, which is mostly anisotropic.


Subject(s)
Apatites , Bone and Bones , Rats , Animals , Apatites/chemistry , Elastic Modulus , Cortical Bone , Bone Density/physiology
20.
J Mech Behav Biomed Mater ; 151: 106364, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38183747

ABSTRACT

Hydroxyapatite (HA) is a non-bioceramic commonly used in human implants in the form of coatings, which are limited in their application by mechanical and wear resistance properties, as well as biodegradability. In this study, fluorine substituted hydroxyapatite (FHA) coatings were prepared on Ti-6Al-4V surfaces by plasma spraying method using a mixture of calcium fluoride and hydroxyapatite powders. The prepared coatings were characterized by X-ray diffraction and fourier transform infrared (FTIR) spectroscopy at different levels of calcium fluoride (3 wt%, 6 wt%, 9 wt%, and 12 wt%). The biocompatibility of the coatings was evaluated by in vitro mineralization experiments. Experimental results showed that at 9 wt% of calcium fluoride, the prepared FHA coatings had better mechanical properties, with improved bond strength (28.2 MPa). The X-ray diffraction patterns of the coatings reflect the fluorine substitution during the spraying process and the 9FHA has the highest crystallinity according to the XRD analysis, which is closely related to the biological activity of the coating. In addition, Potentiodynamic polarisation showed that the sample coated with the 9FHA coating had the highest Ecorr and lowest Icorr, indicating the best corrosion resistance. The FHA coating exhibits faster apatite deposition in simulated body fluid, and the efficiency of apatite deposition increases with the increase of CaF2.


Subject(s)
Apatites , Durapatite , Humans , Durapatite/chemistry , Apatites/chemistry , Fluorine , Corrosion , Calcium Fluoride , Coated Materials, Biocompatible/chemistry , Materials Testing , Surface Properties , Titanium/chemistry , Microscopy, Electron, Scanning , X-Ray Diffraction
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