Aaron Schindeler

Objective: The objective of this review is to evaluate the existing literature on the regenerative potential of recombinant amelogenin, with a particular emphasis on the biological activity, synthesis, and delivery systems relevant to periodontal regeneration. Materials and Methods: A comprehensive literature search was conducted across PubMed, Ovid MEDLINE, Scopus and Web of Science to identify studies on recombinant amelogenin for periodontal regeneration. Inclusion criteria encompassed in vitro and animal model studies reporting on cellular growth, regeneration markers, tissue repair and clinical improvements. Data extraction focused on study design, protein source, delivery system, experimental model, and reported biological outcomes. The review was conducted in accordance with the PRISMA‐ScR guidelines and supplemented by a narrative synthesis of the preclinical literature. Results: The literature search included 23 studies comprising predominantly preclinical cell culture and in vivo mouse studies. Recombinant amelogenin was evaluated across diverse experimental systems and delivery methods, most frequently by direct dissolution in culture media, with fewer studies using injectable hydrogels or scaffold‑based carriers. Although most investigations reported biological activity attributable to recombinant amelogenin, the outcome measures, experimental models, and formulation strategies varied substantially, limiting direct comparison across studies. Conclusion: Recombinant amelogenin shows consistent preclinical activity supporting its biological plausibility as a regenerative agent. However, evidence remains largely preclinical and heterogeneous. Future studies should employ standardised models, optimised delivery systems, and direct comparisons with established therapies to clarify its translational potential in periodontal regeneration.

BACKGROUND: Accurate delineation of scar patterns is valuable for guiding catheter ablation of ventricular tachycardia. We hypothesized that scar and its pattern of distribution can be determined from intracardiac electrograms using computational signal processing and that further improvements in classification can be achieved with a convolutional neural network. METHODS: A total of 5 sheep underwent anteroseptal infarction (plus 1 healthy control) with electroanatomic mapping (129±12 days post-infarct). A whole-heart histological model of the postinfarction scar was created and coregistered to ventricular electrograms. Electrograms were matched to scar pattern categories; no scar, at least endocardial scar: at least intramural scar (intramural scar sparing the endocardium), or epicardial-only scar (epicardial scar sparing the endocardium/intramural space). A suite of signal-processing features was extracted from bipolar electrograms. Furthermore, bipolar and unipolar electrograms were used to train a time series convolutional neural network (InceptionTime). RESULTS: A total of 11 551 electrograms were matched to 451 biopsies. Bipolar and unipolar voltage alone were poor classifiers of scar patterns. For each of the scar labels, 20 bipolar electrogram features (predominantly within the frequency domain) yielded an area under the curve of 0.815, 0.810, 0.704, and 0.681 to predict no scar, at least endocardial scar, at least intramural scar, and epicardial-only scar, respectively. Substantial improvement was achieved with a convolutional neural network trained on unipolar electrograms: areas under the curve and accuracy (averaged across wavefronts) were 0.977 and 0.929 for no scar, 0.970 and 0.919 for at least endocardial scar, 0.909 and 0.959 for at least intramural scar and 0.926 and 0.958 for epicardial-only scar. CONCLUSIONS: Convolutional neural network-derived analysis of unipolar electrogram data has excellent predictive value for determination of scar patterns. Computational analyses of electrogram data beyond voltage and other time-domain features are necessary to improve the identification of arrhythmogenic sites in the ventricle.

Probiotics health benefits are hampered by long-term storage, gastrointestinal transit, and lack of adequate colonization within the colon. To this end, we have designed a core-shell structure that features an acid resistant core formulation with low water activity composed of alginate, hydroxypropyl methyl cellulose, and gellan gum (AHG) and a mucoadhesive shell made from chemically modified carboxymethyl chitosan with polyethylenimine (PEI-CMC). The structure of the core-shell microparticles was examined using scanning electron microscopy, and rheological measurements confirmed the improved ionic interactions between the core and the shell using the PEI-modified CMC. Simulated release from core-shell microparticles using polystyrene beads showed preferential release under intestinal conditions. PEI-CMC coating yielded improvements in mucoadhesion that was consistent with a positive shift in surface charge of the particles. Ex vivo studies using Bifidobacterium lactis probiotic bacteria demonstrated a 1.1 × 10⁵-fold improvement in bacterial viability with encapsulation under storage conditions of high humidity and temperature (30 °C). When exposed to simulated gastric fluid, encapsulation increased the probiotic viability by 3.0 × 10²-fold. In vivo studies utilizing bioluminescent Lactobacillus plantarum in mice revealed that encapsulation extended the duration of the signal within the gut and resulted in higher plate counts in suspensions isolated from the cecum. Conversely, we observed an abrupt loss of signal in the gut of the free probiotic. In conclusion, this core-shell system is suitable for improving probiotic shelf life and maximizing delivery to and retention by the colon.

BACKGROUND: Cardiac magnetic resonance imaging (CMR)–defined ventricular scar and anatomic conduction channels (CMR-CCs) offer promise in delineating ventricular tachycardia substrate. No studies have validated channels with coregistered histology, nor have they ascertained the histological characteristics of deceleration zones (DZs) within these channels. We aimed to validate CMR scar and CMR-CCs with whole-heart histology and electroanatomic mapping in a postinfarction model. METHODS: Five sheep underwent anteroseptal infarction. CMR (116±20 days post infarct) was postprocessed using ADAS-3D, varying pixel intensity thresholds (5545, 6040, 6535, and 7030). DZs were identified by electroanatomic mapping (129±12 days post infarct). Explanted hearts were sectioned and stained with Picrosirius red, and whole-heart histopathologic shells were generated. Scar topography as well as percentage fibrosis, adiposity, and remaining viable myocardium within 3 mm histological biopsies and within CMR-CCs were determined. RESULTS: Using the standard 6040 thresholding, CMR had 83.8% accuracy for identifying histological scar in the endocardium (κ, 0.666) and 61.4% in the epicardium (κ, 0.276). Thirty-seven CMR-CCs were identified by varying thresholding; 23 (62%) were unique. DZs colocalized to 19 of 23 (83%) CMR-CCs. Twenty (87%) CMR-CCs were histologically confirmed. Within-channel histological fibrosis did not differ by the presence of DZs ( P =0.242). Within-channel histological adiposity was significantly higher at sites with versus without DZs (24.1% versus 8.3%; P <0.001). CONCLUSIONS: Postprocessed CMR-derived scars and channels were validated by histology and electroanatomic mapping. Regions of CMR-CCs at sites of DZs had higher adiposity but similar fibrosis than regions without DZs, suggesting that lipomatous metaplasia may contribute to arrhythmogenicity of postinfarction scar.

Neurofibromatosis type 1 (NF1) is a complex genetic disorder that affects a range of tissues including muscle and bone. Recent preclinical and clinical studies have shown that Nf1 deficiency in muscle causes metabolic changes resulting in intramyocellular lipid accumulation and muscle weakness. These can be subsequently rescued by dietary interventions aimed at modulating lipid availability and metabolism. It was speculated that the modified diet may rescue defects in cortical bone as NF1 deficiency has been reported to affect genes involved with lipid metabolism. Bone specimens were analyzed from wild type control mice as well as Nf1_Prx1^-/- (limb-targeted Nf1 knockout mice) fed standard chow versus a range of modified chows hypothesized to influence lipid metabolism. Mice were fed from 4 weeks to 12 weeks of age. MicroCT analysis was performed on the cortical bone to examine standard parameters (bone volume, tissue mineral density, cortical thickness) and specific porosity measures (closed pores corresponding to osteocyte lacunae, and larger open pores). Nf1_Prx1^-/- bones were found to have inferior bone properties to wild type bones, with a 4-fold increase in the porosity attributed to open pores. These measures were rescued by dietary interventions including a L-carnitine + medium-chain fatty acid supplemented chow previously shown to improve muscle histology function. Histological staining visualized these changes in bone porosity. These data support the concept that lipid metabolism may have a mechanistic impact on bone porosity and quality in NF1.

Covalent biofunctionalization of implant surfaces using anti microbial agents is a promising approach to reducing bone infection and implant failure. Radical‐rich, ion‐assisted plasma polymerized (IPP) coatings enable surface covalent biofunctionalization in a simple manner; but until now, they are limited to only 2D surfaces. Here a new technology is demonstrated to create homogenous IPP coatings on 3D materials using a rotating, conductive cage that is negatively biased while immersed in RF plasma. Evidence is provided that under controlled energetic ion bombardment, this technology enables the formation of highly robust and homogenous radical‐rich coatings on 3D objects for subsequent covalent attachment of antimicrobial agents. To functionally apply this technology, the broad‐spectrum antimicrobial CSA‐90 is attached to the surfaces, where it retained potent antibacterial activity against Staphylococcus aureus. CSA‐90 covalent functionalization of stainless‐steel pins used in a murine model of orthopedic infection revealed the highly promising potential of this coating system to reduce S. aureus infection‐related bone loss. This study takes the previous research on plasma‐based covalent functionalization of 2D surfaces a step further, with important implications for ushering in a new dimension in the biofunctionalization of 3D structures for applications in bone implants and beyond.

Senolytics are a category of drugs that reduce the impact of cellular senescence, an effect associated with a range of chronic and age‐related diseases. Since the discovery of the first senolytics in 2015, the number of known senolytic agents has grown dramatically. This review discusses the broad categories of known senolytics—kinase inhibitors, Bcl‐2 family protein inhibitors, naturally occurring polyphenols, heat shock protein inhibitors, BET family protein inhibitors, P53 stabilizers, repurposed anti‐cancer drugs, cardiac steroids, PPAR‐alpha agonists, and antibiotics. The approaches used to screen for new senolytics are articulated including a range of methods to induce senescence, different target cell types, various senolytic assays, and markers. The choice of methods can greatly influence the outcomes of a screen, with high‐quality screens featuring robust systems, adequate controls, and extensive validation in alternate assays. Recent advances in single‐cell analysis and computational methods for senolytic identification are also discussed. There is significant potential for further drug discovery, but this will require additional research into drug targets and mechanisms of actions and their subsequent rigorous evaluation in pre‐clinical models and human trials.

Introduction: Osteomyelitis remains a major clinical challenge. Many published rodent fracture infection models are costly compared with murine models for rapid screening and proof-of-concept studies. We aimed to develop a dependable and cost-effective murine bone infection model that mimics bacterial bone infections associated with biofilm and metal implants. Methods: Tibial drilled hole (TDH) and needle insertion surgery (NIS) infection models were compared in C57BL/6 mice (female, N=150). Metal pins were inserted selectively into the medullary canal adjacent to the defect sites on the metaphysis. Free Staphylococcus aureus (ATCC 12600) or biofilm suspension (ATCC 25923) was locally inoculated. Animals were monitored for physiological or radiographic evidence of infection without prophylactic antibiotics for up to 14 d. At the end point, bone swabs, soft-tissue biopsies, and metal pins were taken for cultures. X-ray and micro-CT scans were performed along with histology analysis. Results: TDH and NIS both achieved a 100 % infection rate in tibiae when a metal implant was present with injection of free bacteria. In the absence of an implant, inoculation with a bacterial biofilm still induced a 40 %–50 % infection rate. In contrast, freely suspended bacteria and no implant consistently showed lower or negligible infection rates. Micro-CT analysis confirmed that biofilm infection caused local bone loss even without a metal implant as a nidus. Although a metal surface permissive for biofilm formation is impermeable to create progressive bone infections in animal models, the metal implant can be dismissed if a bacterial biofilm is used. Conclusion: These models have a high potential utility for modeling surgery-related osteomyelitis, with NIS being simpler to perform than TDH.