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.

This work investigated the range of substitution of two biologically relevant ions, namely Mn2+ and Co2+, into the structure of β-tricalcium phosphate, as well as their influence on bone cells response. To this aim, β-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 β-TCP structure: 15 at% and 20 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 10 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 β-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.

Our previous work (Mol Pharm, 20 (2023) 3427) showed that crystalline excipients, specifically anhydrous dibasic calcium phosphate (DCPA), facilitated the dehydration of carbamazepine dihydrate (CBZDH) and the formation of an amorphous product phase during the mixing stage of continuous tablet manufacturing. Understanding the mechanism of this excipient-induced effect was the object of this study. Blending with DCPA for 15 min caused pronounced lattice disorder in CBZDH. This was evident from the 190% increase in the apparent lattice strain determined by the Williamson-Hall plot. The rapid dehydration was attributed to the increased reactivity of CBZDH caused by this lattice disorder. Lattice disorder in CBZDH was induced by a second method, cryomilling it with DCPA. The dehydration was accelerated in the milled sample. Annealing the cryomilled sample reversed the effect, thus confirming the effect of lattice disorder on the dehydration kinetics. The hardness of DCPA appeared to be responsible for the disordering effect. DCPA exhibited a similar effect in other hydrates, thereby revealing that the effect was not unique to CBZDH. However, its magnitude varied on a case-by-case basis. The high shear powder mixing was necessary for rapid and efficient powder mixing during continuous drug product manufacturing. The mechanical stress imposed on the CBZDH, and exacerbated by DCPA, caused this unexpected destabilization.

Orthodontic relapse is one of the most prevalent concerns of orthodontic therapy. Relapse results in patients\' teeth reverting towards their pretreatment positions, which increases the susceptibility to functional problems, dental disease, and substantially increases the financial burden for retreatment. This phenomenon is thought to be induced by rapid remodeling of the periodontal ligament (PDL) in the early stages and poor bone quality in the later stages. Current therapies, including fixed or removable retainers and fiberotomies, have limitations with patient compliance and invasiveness. Approaches using biocompatible biomaterials, such as calcium phosphate polymer-induced liquid precursors (PILP), is an ideal translational approach for minimizing orthodontic relapse. Here, post-orthodontic relapse is reduced after a single injection of high concentration PILP (HC-PILP) nanoclusters by altering PDL remodeling in the early stage of relapse and improving trabecular bone quality in the later phase. HC-PILP nanoclusters are achieved by using high molecular weight poly aspartic acid (PASP, 14 kDa) and poly acrylic acid (PAA, 450 kDa), which resulted in a stable solution of high calcium and phosphate concentrations without premature precipitation. In vitro results show that HC-PILP nanoclusters prevented collagen type-I mineralization, which is essential for the tooth-periodontal ligament (PDL)-bone interphase. In vivo experiments show that the PILP nanoclusters minimize relapse and improve the trabecular bone quality in the late stages of relapse. Interestingly, PILP nanoclusters also altered the remodeling of the PDL collagen during the early stages of relapse. Further in vitro experiments showed that PILP nanoclusters alter the fibrillogenesis of collagen type-I by impacting the protein secondary structure. These findings propose a novel approach for treating orthodontic relapse and provide additional insight into the PILP nanocluster\'s structure and properties on collagenous structure repair.

The global diffusion of antibiotic resistance poses a severe threat to public health. Addressing antibiotic-resistant infections requires innovative approaches, such as antibacterial nanostructured surfaces (ANSs). These surfaces, featuring ordered arrays of nanostructures, exhibit the ability to kill bacteria upon contact. However, most currently developed ANSs utilize bioinert materials, lacking bioactivity crucial for promoting tissue regeneration, particularly in the context of bone infections. This study introduces ANSs composed of bioactive calcium phosphate nanocrystals. Two distinct ANSs were created through a biomineralization-inspired growth of amorphous calcium phosphate (ACP) precursors. The ANSs demonstrated efficient antibacterial properties against both Gram-negative (P. aeruginosa) and Gram-positive (S. aureus) antibiotic resistant bacteria, with up to 75 % mortality in adhered bacteria after only 4 h of contact. Notably, the ANS featuring thinner and less oriented nano-needles exhibited superior efficacy attributed to simultaneous membrane rupturing and oxidative stress induction. Moreover, the ANSs facilitate the proliferation of mammalian cells, enhancing adhesion, spreading, and reducing oxidative stress. The ANSs displayed also significant bioactivity towards human mesenchymal stem cells, promoting colonization and inducing osteogenic differentiation. Specifically, the ANS with thicker and more ordered nano-needles demonstrated heightened effects. In conclusion, ANSs introduced in this work have the potential to serve as foundation for developing bone graft materials capable of eradicate site infections while concurrently stimulating bone regeneration. STATEMENT OF SIGNIFICANCE: Nanostructured surfaces with antibacterial properties through a mechano-bactericidal mechanism have shown significant potential in fighting antibiotic resistance. However, these surfaces have not been fabricated with bioactive materials necessary for developing devices that are both antibacterial and able to stimulate tissue regeneration. This study demonstrates the feasibility of creating nanostructured surfaces of ordered calcium phosphate nano-needles through a biomineralization-inspired growth. These surfaces exhibit dual functionality, serving as effective bactericidal agents against Gram-negative and Gram-positive antibiotic-resistant bacteria while also promoting the proliferation of mammalian cells and inducing osteogenic differentiation of human mesenchymal stem cells. Consequently, this approach holds promise in the context of bone infections, introducing innovative nanostructured surfaces that could be utilized in the development of antimicrobial and osteogenic grafts.

This study aimed to compare the biological properties of newly synthesized cements based on calcium phosphate with a commercially used cement, mineral trioxide aggregate (MTA). Strontium (Sr)-, Copper (Cu)-, and Zinc (Zn)-doped hydroxyapatite (miHAp) powder was obtained through hydrothermal synthesis and characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy dispersive X-ray spectrometry (EDX). Calcium phosphate cement (CPC) was produced by mixing miHAp powder with a 20 wt.% citric acid solution, followed by the assessment of its compressive strength, setting time, and in vitro bioactivity. Acetylsalicylic acid (ASA) was added to the CPC, resulting in CPCA. Biological tests were conducted on CPC, CPCA, and MTA. The biocompatibility of the cement extracts was evaluated in vitro using human dental pulp stem cells (hDPSCs) and in vivo using a zebrafish model. Antibiofilm and antimicrobial effect (quantified by CFUs/mL) were assessed against Streptococcus mutans and Lactobacillus rhamnosus. None of the tested materials showed toxicity, while CPCA even increased hDPSCs proliferation. CPCA showed a better safety profile than MTA and CPC, and no toxic or immunomodulatory effects on the zebrafish model. CPCA exhibited similar antibiofilm effects against S. mutans and L. rhamnosus to MTA.

Despite years of extensive research, achieving the optimal properties for calcium phosphate-based biomaterials remains an ongoing challenge. Recently, \'biomicroconcretes\' systems consisting of setting-phase-forming bone cement matrix and aggregates (granules/microspheres) have been developed and studied. However, further investigations are necessary to clarify the complex interplay between the synthesis, structure, and properties of these materials. This article focusses on the development and potential applications of hybrid biomaterials based on alpha-tricalcium phosphate (αTCP), hydroxyapatite (HA) and methylcellulose (MC) modified with silver (0.1 wt.% or 1.0 wt.%). The study presents the synthesis and characterization of silver-modified hybrid granules and seeks to determine the possibility and efficiency of incorporating these hybrid granules into αTCP-based biomicroconcretes. The αTCP and hydroxyapatite provide structural integrity and osteoconductivity, the presence of silver imparts antimicrobial properties, and MC allows for the self-assembling of granules. This combination creates an ideal environment for bone regeneration, while it potentially may prevent bacterial colonization and infection. The material\'s chemical and phase composition, setting times, compressive strength, microstructure, chemical stability, and bioactive potential in simulated body fluid are systematically investigated. The results of the setting time measurements showed that both the size and the composition of granules (especially the hybrid nature) have an impact on the setting process of biomicroconcretes. The addition of silver resulted in prolonged setting times compared to the unmodified materials. Developed biomicroconcretes, despite exhibiting lower compressive strength compared to traditional calcium phosphate cements, fall within the range of human cancellous bone and demonstrate chemical stability and bioactive potential, indicating their suitability for bone substitution and regeneration. Further in vitro studies and in vivo assessments are needed to check the potential of these biomaterials in clinical applications.

Carbonated apatite (CAp), known as the main mineral that makes up human bone, can be utilized in conjunction with scaffolds to increase their bioactivity. Various methods (e.g., co-precipitation, hydrothermal, and biomimetic coatings) have been used to provide bioactivity by forming CAp on surfaces similar to bone minerals. Among them, the use of simulated body fluids (SBF) is the most popular biomimetic method for generating CAp, as it can provide a mimetic environment. However, coating methods using SBF require at least a week for CAp formation. The long time it takes to coat biomimetic scaffolds is a point of improvement in a field that requires rapid regeneration. Here, we report a step-wise biomimetic coating method to form CAp using calcium carbonate vaterite (CCV) as a precursor. We can manufacture CCV-transformed CAp (V-CAp) on the surface in 4 h at least by immersing CCV in a phosphate solution. The V-CAp deposited surface was analyzed using scanning electron microscopy (SEM) images according to the type of phosphate solutions to optimize the reaction conditions. X-ray diffraction (XRD) and attenuated total reflection-Fourier transform infrared (ATR-FTIR) analysis validated the conversion of CCV to V-CAp on surfaces. In addition, the bioactivity of V-CAp coating was analyzed by the proliferation and differentiation of osteoblasts in vitro. V-CAp showed 2.3-folded higher cell proliferation and 1.4-fold higher ALP activity than the glass surface. The step-wise method of CCV-transformed CAp is a biocompatible method that allows the environment of bone regeneration and has the potential to confer bioactivity to biomaterial surfaces, such as imparting bioactivity to non-bioactive metal or scaffold surfaces within one day. It can rapidly form carbonated apatite, which can greatly improve time efficiency in research and industrial applications.

The skin is the outermost part of the body. Although susceptible to damage, the skin is in direct contact with the external environment. Wound dressing is a clinical method that plays a vital role in wound healing. Herein, we developed an antibacterial wound dressing using alginate as the basic material. The dressing was prepared using the solvent casting method, which was used to analyze the effects of adding CaP and ZnO on its structural, optical, and antibacterial properties. Adding CaP exhibited strong but stiff mechanical properties, unlike the CaP/ZnO, which possessed high strength and elasticity. The optical properties of sample S2 did not have a considerable impact. By contrast, the addition of ZnO to sample S3 notably increases the wavelength and absorption value. The diameter of the inhibition zone for S. aureus bacteria exhibited a successive increase in its antibacterial properties, and sample S3 exhibited the highest value. Thus, sample S3 is the most promising wound dressing concerning speeding up the wound healing process because it possesses the most optimal mechanical, optical, and antibacterial properties. The main limitation to be addressed is that sample S3 cannot be easily digested in the environment.

The conventional treatment of osteomyelitis with antibiotic-loaded nondegradable polymethylmethacrylate (ATB-PMMA) beads has certain limitations, including impeded bone reconstruction and the need for secondary surgery. To overcome this challenge, this study aimed to develop and characterize an injectable vancomycin-loaded silk fibroin/methylcellulose containing calcium phosphate-based in situ thermosensitive hydrogel (VC-SF/MC-CAPs). The VC-SF/MC-CAPs solution can be easily administered at room temperature with a low injectability force of ≤30 N and a high vancomycin (VC) content of ~96%. Additionally, at physiological temperature (37 °C), the solution could transform into a rigid hydrogel within 7 minutes. In vitro drug release performed under both physiological (pH 7.4) and infection conditions (pH 4.5) revealed a prolonged release pattern of VC-SF/MC-CAPs following the Peppas-Sahlin kinetic model. In addition, the released VC from VC-SF/MC-CAPs hydrogels exhibited antibacterial activity against Staphylococcus aureus for a period exceeding 35 days, as characterized by the disk diffusion assay. Furthermore, at pH 7.4, the VC-SF/MC-CAPs demonstrated >60% degradation within 35 days. Importantly, when exposed to physiological pH conditions, CAPs are transformed into bioactive hydroxyapatite, which benefits bone formation. Therefore, VC-SF/MC-CAPs showed significant potential as a local drug delivery system for treating osteomyelitis.