A bifunctional lactoferrin-derived amyloid coating prevents bacterial adhesion and occludes dentinal tubules via deep remineralization.

Dentin hypersensitivity (DH) manifests as sharp and uncomfortable pain due to the exposure of dentinal tubules (DTs) following the erosion of tooth enamel. Desensitizing agents commonly used in clinical practice have limitations such as limited depth of penetration, slow remineralization and no antimicrobial properties. To alleviate these challenges, our study designed a lactoferrin-derived amyloid nanofilm (PTLF nanofilm) inspired by the saliva-acquired membrane (SAP). The nanofilm utilises Tris(2-carboxyethyl)phosphine (TCEP) to disrupt the disulfide bonds of lactoferrin (LF) under physiological conditions. The PTLF nanofilm modifies surfaces across various substrates and effectively prevents the early and stable adhesion of cariogenic bacteria, such as Streptococcus mutans and Lactobacillus acidophilus. Simultaneously, it adheres rapidly and securely to demineralized dentin surfaces, facilitating in-situ remineralization of HAP through a simple immersion process. This leads to the formation of a remineralized layer resembling natural dentin, with an occlusion depth of dentinal tubules exceeding 80 µm after three days. The in vivo and vitro results confirm that the PTLF nanofilm possesses good biocompatibility and its ability to exert simultaneous antimicrobial effects and dentin remineralization. Accordingly, this innovative bifunctional PTLF amyloid coating offers promising prospects for the management of DH-related conditions. STATEMENT OF SIGNIFICANCE.

A biomimetic piezoelectric scaffold with sustained Mg2+ release promotes neurogenic and angiogenic differentiation for enhanced bone regeneration.

Natural bone is a composite tissue made of organic and inorganic components, showing piezoelectricity. Whitlockite (WH), which is a natural magnesium-containing calcium phosphate, has attracted great attention in bone formation recently due to its unique piezoelectric property after sintering treatment and sustained release of magnesium ion (Mg2+). Herein, a composite scaffold (denoted as PWH scaffold) composed of piezoelectric WH (PWH) and poly(ε-caprolactone) (PCL) was 3D printed to meet the physiological demands for the regeneration of neuro-vascularized bone tissue, namely, providing endogenous electric field at the defect site. The sustained release of Mg2+ from the PWH scaffold, displaying multiple biological activities, and thus exhibits a strong synergistic effect with the piezoelectricity on inhibiting osteoclast activation, promoting the neurogenic, angiogenic, and osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSCs) in vitro. In a rat calvarial defect model, this PWH scaffold is remarkably conducive to efficient neo-bone formation with rich neurogenic and angiogenic expressions. Overall, this study presents the first example of biomimetic piezoelectric scaffold with sustained Mg2+ release for promoting the regeneration of neuro-vascularized bone tissue in vivo, which offers new insights for regenerative medicine.

Injectable, High Specific Surface Area Cryogel Microscaffolds Integrated with Osteoinductive Bioceramic Fibers for Enhanced Bone Regeneration.

Organic-inorganic composites with high specific surface area and osteoinductivity provide a suitable microenvironment for cell ingrowth and effective ossification, which could greatly promote bone regeneration. Here, we report gelatin methacryloyl (GelMA) cryogel microspheres that are reinforced with hydroxyapatite (HA) nanowires and calcium silicate (CS) nanofibers to achieve the goal. The prepared composite cryogel microspheres with open porous structure and rough surface greatly facilitate cell anchoring, simultaneously exhibiting excellent injectability. Compared to the only HA- or CS-containing counterparts, the GelMA cryogel microspheres composited with HA:CS (termed as GMHC) achieve sustained release of bioactive Ca, P, and Si elements, which are conducive to osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSCs). These composite microspheres can prevent from forming peralkalic conditions, which is beneficial for cell growth. After injection of cryogel microspheres into rat calvarial defects, neo-bone tissue grows into their pores, showing tight integration. The embedded bioceramic components significantly promote bone regeneration, with the GMHC achieving the best regenerative outcomes. Promisingly, porous organic-inorganic composite cryogel microspheres, with high specific surface area, biodegradability, and osteoinductivity, can act as injectable microscaffolds to repair bone defects with enhanced efficiency, which may widen the scaffold strategy for bone tissue engineering.

Amyloid-Mediated Nanoarchitectonics with Biomimetic Mineralization of Polyetheretherketone for Enhanced Osseointegration.

Polyetheretherketone (PEEK), a widely used implant material, has attracted the attention of scientific researchers because of its bone-matched elastic modulus, radiolucency, and chemical resistance. However, the bioinert chemical properties of PEEK do not promote bone apposition once implanted. In this study, using a phase-transitioned lysozyme (PTL) nanofilm as a sandwiched layer, a robust hydroxyapatite (HAp) coating on PEEK (HAp@PTL@PEEK) is constructed. The PTL nanofilm shows strong adhesion to the PEEK surface and induces biomimetic mineralization to form a compact HAp coating on PEEK in simulated body fluids. This HAp coating not only shares a higher adhesion strength and better stability but can also be applied to implants with complex 3D structures. HAp@PTL@PEEK showed significantly enhanced osteogenic capacity when cultured with rat bone marrow mesenchymal stem cells by promoting initial cell adhesion, proliferation, and osteogenic differentiation in vitro. In vivo evaluations utilizing models of femoral condyle defects and skull defects confirm that the HAp coating substantially augments bone remodeling and osseointegration ability. Compared with the traditional method, our modified method is simpler, more environmentally friendly, and uses less hazardous components. Furthermore, the obtained HAp coating shares a higher adhesion strength to PEEK and a better osteogenic capacity. The study offers a novel method to improve the osseointegration of PEEK-based implants in biointerfaces and tissue engineering.

Integrated Piezoelectric/Conductive Composite Cryogel Creates Electroactive Microenvironment for Enhanced Bone Regeneration.

Natural bone tissue possesses inherent electrophysiological characteristics, displaying conductivity and piezoelectricity simultaneously; hence, the reconstruction of local electrical microenvironment at defect site provides an effective strategy to enhance osteogenesis. Herein, a composite cryogel-type scaffold (referred to as Gel-PD-CMBT) is developed for bone regeneration, utilizing gelatin (Gel) in combination with a conductive poly(ethylene dioxythiophene)/polystyrene sulfonate matrix and Ca/Mn co-doped barium titanate (CMBT) nanofibers as the piezoelectric filler. The incorporation of these components results in the formation of an integrated piezoelectric/conductive network within the scaffold, facilitating charge migration and yielding a conductivity of 0.59 S cm-1 . This conductive scaffold creates a promising electroactive microenvironment, which is capable of up-regulating biological responses. Furthermore, the interconnected porous structure of the Gel-PD-CMBT scaffold not only provides mechanical stability but also offered ample space for cellular and tissue ingrowth. This Gel-PD-CMBT scaffold demonstrates a greater capacity to promote cellular osteogenic differentiation in vitro and neo-bone formation in vivo. In summary, the Gel-PD-CMBT scaffold, with its integrated piezoelectricity and conductivity, effectively restores the local electroactive microenvironment, offering an ideal platform for the regeneration of electrophysiological bone tissue.

Polyzwitterion Manipulates Remineralization and Antibiofilm Functions against Dental Demineralization.

Biomineralization technology has become a trend for the arrest and prevention of dental caries. In particular, the bioactivity and ability to release large amounts of Ca2+ and PO43- ions make amorphous calcium phosphate (ACP) for hard tissue remineralization are highly desired. However, the instability of ACP limits its clinical application. Under continuous bacterial challenge in the oral cavity, the currently developed ACP-based remineralization system lacks the ability to inhibit bacterial adhesion and biofilm formation. Here, a dual-functional nanocomposite with antibiofilm and remineralization properties was designed by combining zwitterionic poly(carboxybetaine acrylamide) (PCBAA) and ACP. The resulting nanocomposite was stable in solution for at least 3 days without any aggregation. The PCBAA/ACP nanocomposite exerted a significant inhibitory effect on the adhesion and biofilm formation of Streptococcus mutans and exhibited bactericidal activities under acidic conditions resulting from bacteria. Moreover, compared with fluoride, this nanocomposite demonstrated superior effects in promoting the remineralization of demineralized enamel and the occlusion of exposed dentinal tubules in vivo and in vitro. The present work provides a theoretical and experimental basis for the use of the PCBAA/ACP nanocomposite as a potential dual-functional agent for arresting and preventing caries.

In vitroandin vivoevaluation of biomimetic hydroxyapatite/whitlockite inorganic scaffolds for bone tissue regeneration.

Native bone tissue can be formed by developing collagen fibrils coated with hydroxyapatite (HA) and whitlockite (WH) nanoparticles after mineralization. WH has attracted much attention as the second most abundant bone mineral in human bones. It has a negatively charged surface, which can adsorb osteogenesis-related proteins such as bone sialoproteinin vivo, thus having a stronger possibility to induce osteogenesis. However, due to its poor thermodynamic stability and intermediate phases, the preparation of WH is relatively tricky, so WH inorganic scaffolds are still rarely studied. Therefore, this study explored the preparation of WH inorganic scaffolds using the hydrothermal method and prepared pure inorganic WH scaffolds. The prepared scaffolds exhibited apparent WH crystal phases in the x-ray powder diffraction (XRD) characterization. In the scanning electron microscopy (SEM) images, the WH scaffolds had an apparent hexagonal crystal form, which had a pronounced effect on promoting cell proliferation and differentiationin vitroexperiments compared to the HA and HA/WH scaffolds. Furthermore, the scaffolds were used to verify the osteogenic properties of subcutaneous ectopic osteogenesis or repair of the calvarial defectin vivoand proved that the WH inorganic scaffolds have an excellent synergistic osteogenic ability.

Doping bioactive elements into a collagen scaffold based on synchronous self-assembly/mineralization for bone tissue engineering.

Pure collagen is biocompatible but lacks inherent osteoinductive, osteoimmunomodulatory and antibacterial activities. To obtain collagen with these characteristics, we developed a novel methodology of doping bioactive elements into collagen through the synchronous self-assembly/mineralization (SSM) of collagen. In the SSM model, amorphous mineral nanoparticles (AMN) (amorphous SrCO3, amorphous Ag3PO4, etc.) stabilized by the polyampholyte, carboxymethyl chitosan (CMC), and collagen molecules were the primary components under acidic conditions. As the pH gradually increased, intrafibrillar mineralization occurred via the self-adaptive interaction between the AMNs and the collagen microfibrils, which were self-assembling; the AMNs wrapped around the microfibrils became situated in the gap zones of collagen and finally transformed into crystals. Sr-doped collagen scaffolds (Sr-CS) promoted in vitro cell proliferation and osteogenic differentiation of rat bone marrow mesenchymal stromal cells (rBMSCs) and synergistically improved osteogenesis of rBMSCs by altering the macrophage response. Ag-doped collagen scaffolds (Ag-CS) exhibited in vitro antibacterial effects on S. aureus, as well as cell/tissue compatibility. Moreover, Sr-CS implanted into the calvarial defect of a rat resulted in improved bone regeneration. Therefore, the SSM model is a de novo synthetic strategy for doping bioactive elements into collagen, and can be used to fabricate multifunctional collagen scaffolds to meet the clinical challenges of encouraging osteogenesis, boosting the immune response and fighting severe infection in bone defects.