Calcified Nodules Research Articles

LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment

BackgroundInflammation often causes irreversible damage to dental pulp tissue. Dental pulp stem cells (DPSCs), which have multidirectional differentiation ability, play critical roles in the repair and regeneration of pulp tissue. However, the presence of proinflammatory factors can affect DPSCs proliferation, differentiation, migration, and other functions. LL-37 is a natural cationic polypeptide that inhibits lipopolysaccharide (LPS) activity, enhances cytokine production, and promotes the migration of stem cells. However, the potential of LL-37 in regenerative endodontics remains unknown. This study aimed to investigate the regulatory role of LL-37 in promoting the migration and odontogenic differentiation of DPSCs within an inflammatory microenvironment. These findings establish an experimental foundation for the regenerative treatment of pulpitis and provide a scientific basis for its clinical application.Materials and methodsDPSCs were isolated via enzyme digestion combined with the tissue block adhesion method and identified via flow cytometry. The impact of LL-37 on the proliferation of DPSCs was evaluated via a CCK-8 assay. The recruitment of DPSCs was assessed through a transwell assay. The mRNA expression levels of inflammatory and aging-related genes were assessed via reverse transcription‒polymerase chain reaction (RT‒PCR), western blotting, and enzyme‒linked immunosorbent assay (ELISA). The odontogenic differentiation of DPSCs was assessed through alkaline phosphatase (ALP) staining, alizarin red staining, and RT‒PCR analysis.ResultsLL-37 has the potential to enhance the migration of DPSCs. In an inflammatory microenvironment, LL-37 can suppress the expression of genes associated with inflammation and aging, such as TNF-α, IL-1β, IL-6, P21, P38 and P53. Moreover, it promotes odontogenic differentiation in DPSCs by increasing ALP activity, increasing calcium nodule formation, and increasing the expression of dentin-related genes such as DMP1, DSPP and BSP.ConclusionThese findings suggest that the polypeptide LL-37 facilitates the migration of DPSCs and plays a crucial role in resolving inflammation and promoting cell differentiation within an inflammatory microenvironment. Consequently, LL-37 has promising potential as an innovative therapeutic approach for managing inflammatory dental pulp conditions.

Outcomes of coronary intravascular lithotripsy for the treatment of calcified nodules: a pooled analysis of the Disrupt CAD studies.

Coronary intravascular lithotripsy (IVL) safely facilitates stent implantation in severely calcified lesions. This analysis sought to determine the relative impact of IVL on acute and long-term outcomes specifically in calcified nodules (CNs). Individual patient-level data (N=155) were pooled from the Disrupt CAD optical coherence tomography (OCT) substudies. Severely calcified lesions with and without CNs were compared by OCT for acute procedural results and for target lesion failure (TLF) at 2 years - a composite of cardiac death, target vessel myocardial infarction, and ischaemia-driven target lesion revascularisation. A CN was identified in 18.7% (29/155) of lesions. When comparing lesions with and without CNs, there were no significant differences in preprocedure minimal lumen area or diameter stenosis; however, the mean calcium angle and calcium volume were greater in CN lesions. Despite a higher calcium burden, the final minimal stent area (CN: 5.7 mm2 [interquartile range [IQR] 4.4, 8.3] vs non-CN: 5.7 mm2 [IQR 4.7, 7.2]; p=0.80) and stent expansion (CN: 79.3% [IQR 64.3, 87.0] vs 80.2% [IQR 68.9, 92.4]; p=0.30) were comparable between the two groups. In the CN group, the final stent area and expansion at CN sites were 7.6 mm2 (IQR 5.5, 8.5) and 89.7% (IQR 79.8, 102.5), respectively. The cumulative incidence of TLF at 2 years was 13.9% and 8.0% in the CN and non-CN groups, respectively (p=0.32). Despite a greater calcium volume in CNs, IVL use was associated with comparable stent expansion and luminal gain in both CN and non-CN lesions. Further studies powered for clinical outcomes comparing different plaque modification techniques in this lesion subset are warranted.

Numerical investigation of the effects of pre-dilatation on paravalvular leakage during transcatheter aortic valve implantation

Abstract Due to promising results, the patient cohort for transcather aortic valve replacement (TAVR) has been extended in recent years to include patients with a bicuspid aortic valve (BAV). There are different types of BAV. One variant is the tricommisural bicuspid aortic valve (TBAV). BAV have an increased risk of post-TAVR complications such as paravalvular leakage. In the case of paravalvular leakage, blood flows past the prosthesis back into the ventricle during diastole. Clinically, patients with BAV are often pre-dilated. For this reason, we want to investigate how pre-dilatation of BAV can affect the leakage rate. A simplified model is used for pre-dilatation, where the calcification nodule is cut along the free edge of the leaflets before the deployment. In order to evaluate the effects of this method, a deployment simulation was carried out for both geometries using an explicit calculation. A flow simulation was then performed to determine the paravalvular leakage. The pre-dilatation allows the leaflets to move independently of each other. Without pre-dilatation, the TAVR cannot fully expanded. The leakage rate is higher for the BAV than for the pre-dilated geometry (53.1mLs−1 vs. 19.4mLs−1). In this model, we have shown the effect of pre-dilatation on implantation results.

Multi-type classification of lung nodules based on CT radiomics and ensemble learning for diversity weighting.

The accurate classification of lung nodules is critical to achieving personalized lung cancer treatment and prognosis prediction. The treatment options for lung cancer and the prognosis of patients are closely related to the type of lung nodules, but there are many types of lung nodules, and the distinctions between certain types are subtle, making accurate classification based on traditional medical imaging technology and doctor experience challenging. This study adopts a novel approach, using computed tomography (CT) radiomics to analyze the quantitative features in CT images to reveal the characteristics of lung nodules, and then employs diversity-weighted ensemble learning to enhance the accuracy of classification by integrating the predictive results of multiple models. We extracted lung nodules from the Lung Image Database Consortium image collection (LIDC-IDRI) dataset and derived radiomics features from the nodules. For the classification tasks of seven types of lung nodules, each was split into binary classifications. Two model-building methods were employed: M1 (equal-weighted voting ensemble classifier) and M2 (diversity-weighted voting ensemble classifier). Models were evaluated using 10-fold cross-validation with metrics including the area under the receiver operating characteristic curve (AUC), accuracy, specificity, and sensitivity. Both methods effectively completed classification tasks. The M2 method outperformed M1, particularly in classifying texture, calcification, and the benign and malignant nature of lung nodules. The AUC values of the M2 method in the four subclassifications of texture types of lung nodules were 0.9913, 0.8838, 0.9525, and 0.8845, with the corresponding accuracies of 0.9651, 0.8116, 0.9000, and 0.8284, respectively. In the classification of the degree of calcification of lung nodules, the AUC value of the M2 method was 0.9775 with an accuracy of 0.9642. In the classification of the benign and malignant nature of lung nodules, the AUC value of the M2 method was 0.8953 with an accuracy of 0.8168. The combination of CT radiomics and diversity-weighted ensemble learning effectively identifies lung nodule types, providing a novel method for the precise classification of lung nodules and aiding personalized lung cancer treatment and prognosis prediction. The combination of CT radiomics and ensemble learning for diversity weighting can be well realized to identify the type of lung nodules.

Endoscopic Endonasal Approach for Calcified Sellar/Parasellar Region Pathologies: Report of 11 Pituitary Adenoma Cases

Calcification in pituitary adenomas is a rare occurence and its differential diagnosis typically includes other sellar masses.Common calcifications in pituitary adenomas are classified into two morphological forms: capsular(eggshell-like)and multiple small nodular calcifications located within the adenoma.Also,there is a pituitary stone term.This study aims to present the results of calcified pituitary adenoma case series who underwent endoscopic endonasal approach(EEA)and the clinical,histopathological characteristics and surgical outcomes of these cases. This study conducted a retrospective cohort analysis of patients with calcified pituitary adenoma operated on by EEA between August 1997 and February 2024.The inclusion criteria were as follows:proven radiological calcification on pre-operative neuroimaging,intraoperative findings of calcification and definitive histopathological diagnosis of calcification.Among these patients,11 cases were included. The mean follow-up duration was 51.45±37.24 (6-118)months.Based on the pre-operative paranasal sinus computed tomography(CT)scans of patients,nine patients(81.8%)had intratumoral calcification,one(9.1%)had capsular(eggshell like)calcification and one(9.1%)had both intratumoral and capsular calcification.Moreover,stone-like calcifications were observed in four patients(36.4%),soft type calcifications in three(27.3%),hard-type calcifications in three(27.3%)and soft and hard-type calcifications in one(9%).Gross total resection(GTR)was achieved in nine patients(81.8%).Pathologic subtypes included non-functioning(n = 4),prolactin secreting(n = 3),growth hormone secreting adenoma(n = 2)and pituitary apoplexy(n = 2).Ten patients had psammomatous type calcifications and one had extensive ossification and osteoid metaplasia. Pre-operative radiological evaluation,intraoperative classification of calcification,and post-operative histopathological assessments are crucial in the treatment of calcified adenomas.Bases on these findings,the EEA,with its advantages,is an approach that can be effectively used in the management of these calcified adenomas.

Transglutaminase-triggered dual gradients of mechanical and biochemical cues self-assembling peptide hydrogel for guiding MC3T3-E1 cell behaviors

The mechanical properties and bioactive motif densities of extracellular matrix materials play crucial roles in regulating cell behaviors, such as cell adhesion, migration, proliferation, and differentiation. However, current studies on cellular responses to ECM predominantly concentrated on polymer hydrogels featuring a single factor, such as the mechanical strength, the types of bioactive motifs, and the morphology of the polymers. This limited focus may overlook the complex interplay of multiple factors. Here, we developed dual gradient peptide Q3GT-I3K hydrogels with tunable mechanical strength (0.3–4.0 kPa) and different density of bioactive motif (0.45–3.67 mM) by enzymatic crosslinking. These hydrogels can mimic the viscoelasticity of natural soft tissues. The properties of mechanical strength and cell responsive motif density could be controlled by modulating the proportion of the substrates in the enzymatic reaction. MC3T3 cells significantly differentiated into osteoblasts after seeded on the Q3GT-I3K hydrogel (2.8 kPa, 1.83 mM Q3GT) for 21 days, identifying from the elevated expression of alkaline phosphatase and substantial calcium nodule formation. Importantly, the engineered hydrogels exert a synergistic effect on the cell behaviors such as early adhesion, late proliferation, and differentiation of MC3T3-E1 cells. This paper introduces a new strategy for designing tissue engineering scaffold materials with specific functions.

Contrast-enhanced US Bosniak Classification: intra- and inter-rater agreement, confounding features, and diagnostic performance

BackgroundThe contrast-enhanced US (CEUS) Bosniak classification, proposed by the European Federation for Ultrasound in Medicine and Biology (EFSUMB) in 2020, predicts malignancy in cystic renal masses (CRMs). However, intra- and inter-rater reproducibility for CEUS features has not been well investigated.PurposeTo explore intra- and inter-rater agreement for US features, identify confounding features, and assess the diagnostic performance of CEUS Bosniak classification.Materials and methodsThis retrospective study included patients with complex CRMs who underwent CEUS examination from January 2013 to August 2023. Radiologists (3 experts and 3 novices) evaluated calcification, echogenic content, wall, septa, and internal nodules of CRMs using CEUS Bosniak classification. Intra- and inter-rater agreements were assessed using the Gwet agreement coefficient (Gwet’s AC). Linear regression identified features associated with discrepancies in Bosniak category assignment. Diagnostic performance was evaluated using the area under the receiver operating characteristic curve (AUC).ResultsA total of 103 complex CRMs were analyzed in 103 patients (mean age, 50 ± 15 years; 66 males). Intra-rater agreement for the Bosniak category was substantial to almost perfect (Gwet’s AC 0.73–0.87). Inter-rater agreement was substantial for the Bosniak category (Gwet’s AC 0.75) and moderate to almost perfect for US features (Gwet’s AC 0.44–0.94). Nodule variation (i.e., absence vs. obtuse margin vs. acute margin) explained 84% of the variability in the Bosniak category assignment. CEUS Bosniak classification showed good diagnostic performance, with AUCs ranging from 0.78 to 0.90 for each rater.ConclusionsCEUS Bosniak classification demonstrated substantial intra- and inter-rater reproducibility and good diagnostic performance in predicting the malignancy potential of CRMs. Nodule variations significantly predicted differences in Bosniak category assignments.Critical relevance statementContrast-enhanced US Bosniak classification reliably predicts malignancy in cystic renal masses, demonstrating substantial reproducibility and diagnostic accuracy. This improves clinical decision-making and patient management.Key PointsIntra- and inter-rater reproducibility for contrast-enhance US features for Bosniak classification have not been well investigated.Substantial inter-rater agreements for the Bosniak category and variable agreements for determining imaging features were found.Contrast-enhanced US Bosniak classification is reproducible and has good diagnostic performance for predicting malignancy in cystic renal masses.Graphical

Mechanism of MiR-145a-3p/Runx2 pathway in dexamethasone impairment of MC3T3-E1 osteogenic capacity in mice.

In this experiment, we screened key miRNAs involved in the dexamethasone-induced decrease in osteogenic capacity of mouse precursor osteoblasts MC3T3-E1 over and investigated their specific regulatory mechanisms. In this experiment, cell counting kit assay was utilized to act on MC3T3-E1 cells at 0, 5μM, 10μM, 15μM concentrations of dexamethasone for 24h, 48h and 72h to observe the changes in cell viability in order to select the appropriate dexamethasone concentration. Apoptosis and reactive oxygen species were detected by flow cytometry. The transcription of osteogenesis-related genes (Runx2, ALP, OCN, OPN, OPG, COL1A1) and protein expression levels (Runx2, ALP, OCN, OPN) were detected by Western Blot and qRT-PCR to validate the changes in cellular osteogenesis. The differentially expressed miRNAs related to MC3T3-E1 osteogenic differentiation after dexamethasone action were screened out. The expression levels of selected target miRNAs were verified in the experimental group and the control group by qRT-PCR. The miRNA inhibitor was transfected to knock down miRNA in dexamethasone-induced MC3T3-E1 injury. Alkaline phosphatase staining and flow cytometry were performed to detect apoptosis and reactive oxygen species changes. transcript and protein expression levels of osteogenesis-related genes in mouse MC3T3-E1 were detected by qRT-PCR and Western blot experiments. By miRNA target gene prediction, luciferase reporter gene experiments, qRT-PCR and Western blot experiments were used to verify whether the selected target miRNAs targeted the target gene. First, it was determined that 10μM dexamethasone solution was effective in inducing a decrease in osteogenic function in mouse MC3T3-E1 by CCK8 experiments, which showed a significant decrease in alkaline phosphatase activity, a decrease in calcium nodules as shown by alizarin red staining, an increase in apoptosis and reactive oxygen species as detected by flow cytometry, as well as a decrease in the expression of osteogenesis-related genes and proteins. Five target miRNAs were identified: miR-706, miR-296-3p, miR-7011-5p, miR-145a-3p, and miR-149-3p. miR-145a-3p, which had the most pronounced and stable expression trend and was the most highly expressed miRNA, was chosen as the target of this experiment by qRT-PCR analysis. -145a-3p, as the subject of this experiment. Knockdown of miR-145a-3p in MC3T3-E1 cells after dexamethasone action significantly improved the expression of their impaired osteogenic indicators. It was shown that after knocking down the target miRNA, alkaline phosphatase staining was significantly increased compared with the dexamethasone-stimulated group and approached the level of the blank control group. Meanwhile, the expression of osteogenic function-related proteins and genes also increased in the dexamethasone-stimulated group after knocking down miR-145a-3p, and approached the level of the blank control group. A direct targeting relationship between miR-145a-3p and Runx2 was indeed confirmed by luciferase reporter gene assays, qRT-PCR and Western blot experiments. The results indicated that dexamethasone impaired the osteogenic differentiation ability of MC3T3-E1 cells by inducing the up-regulation of miR-145a-3p expression. MiR-145a-3p inhibited the osteogenic differentiation ability of MC3T3-E1 cells by targeting and suppressing the expression level of Runx2 protein. Inhibition of miR-145a-3p levels significantly improved the osteogenic differentiation ability of MC3T3-E1 cells.

Natural history of a newly developed calcified nodule: incidence, predictors, and clinical outcomes.

Calcified nodules (CNs) are an increasingly important, high-risk lesion subset. We sought to identify the emergence of new CNs and the relation between underlying plaque characteristics and new CN development. Patients who had undergone two optical coherence tomography (OCT) studies that imaged the same untreated calcified lesion at baseline and follow-up were included. New CNs were an accumulation of small calcium fragments at follow-up that were not present at baseline. Cardiac death, myocardial infarction (MI), or clinically driven revascularisation related to OCT-imaged, but untreated, calcified lesions were then evaluated. Among 372 untreated calcified lesions, with a median of 1.5 (first and third quartiles: 0.7-2.9) years between baseline and follow-up OCTs, new CNs were observed in 7.0% (26/372) of lesions at follow-up. Attenuated calcium representing residual lipid (odds ratio [OR] 3.38, 95% confidence interval [CI]: 1.15-9.98; p=0.03); log10 calcium volume index (length×maximum arc×maximum thickness; OR 2.76, 95% CI: 1.10-6.95; p=0.03); angiographic Δangle between systole and diastole, per 10° (OR 2.30, 95% CI: 1.25-4.22; p=0.01); and time since baseline OCT, per year (OR 1.36, 95% CI: 1.05-1.75; p=0.02) were all associated with new CN development. Clinical events were revascularisation and/or MI and were more frequent in lesions with versus without a new CN (29.3% vs 15.3%; p=0.04). New CNs developed in untreated, lipid-containing, severely calcified lesions with a larger angiographic hinge motion (between systole and diastole), compared with lesions without CNs, and were associated with worse clinical outcomes.

Calcium Phosphate Loaded with Curcumin Prodrug and Selenium Is Bifunctional in Osteosarcoma Treatments.

Although SeO32- ions have been loaded onto calcium phosphate to treat a wide range of cancers, the quest to promote bone tissue regeneration is still ongoing. Curcumin (cur), an herbal extraction, can selectively inhibit tumor cells and promote osteogenesis. In this study, SeO32- ions were co-precipitated in biomimetic calcium phosphate (Se@BioCaP), and modified curcumin prodrug (mcur) was adsorbed on diverse Se@BioCaP surfaces (mcur-Se@BioCaP-Ads). Co-precipitation yielded Se@BioCaP with a significantly higher Se content and exhibited a tailorable micro-/nanostructure. The favorable pH-responsive release of Se and mcur from mcur-Se@BioCaP-Ads showed a synergistic anticancer efficiency in OS cells, enhancing OS cell inhibition more than a single dose of them, which might be associated with ROS production in OS cells. In addition, increased alkaline phosphatase activity and calcium nodule formation in MC3T3-E1 pre-osteoblasts were also verified. These results suggest this novel mcur-Se@BioCaP-Ads has promising and widespread potential in OS treatments.

Morphological characterization of coronary plaques in young indian patients with acute coronary syndrome: A multicentric study

ObjectivesThe prevalence of atherosclerosis and acute coronary syndrome (ACS) is increasing in young Indians (18–50 years of age). However, the characteristics of atherosclerotic plaques in such individuals are poorly understood, presenting distinct challenges for the management of ACS. This study aims to analyze plaque characteristics in young Indian patients with ACS who underwent percutaneous coronary intervention (PCI) using optical coherence tomography (OCT) imaging. MethodsThis was a prospective, multicentric, non-interventional study on patients aged 18–50 years presenting with ST-elevation myocardial infarction (STEMI), non-ST elevation myocardial infarction, or unstable angina, and were scheduled to undergo OCT-guided PCI. Major adverse cardiac events (MACE) were assessed post-procedure and at the 6-month and 12-month follow-ups. ResultsThe study included 100 ACS patients (mean age = 43.6 ± 5.2 years), with 51% presenting with STEMI. Pre-PCI OCT assessment showed that fibrous plaques (75%) were most common followed by plaques containing macrophages (27%), microchannels (20%), and calcified nodules (14%). In addition, plaque rupture, plaque erosion, and lipid-rich plaques, along with red, white, and mixed thrombi, were observed in 31%, 25%, 24%, 21%, 14%, and 17% (total thrombus occurrence = 52%) of the patients, respectively. At 12 months, the MACE (coronary artery bypass graft) rate was 1%. ConclusionsYoung Indian patients with ACS displayed a range of plaque morphologies identified through pre-PCI OCT. Among these, fibrous plaques were the most prominent type, followed by plaques containing macrophages. Additionally, plaque rupture, plaque erosion, and lipid-rich plaques were also observed in this population.

Exploring the osteogenic effects of simiao wan through activation of the PI3K/AKT pathway in osteoblasts

Ethnopharmacological relevanceOsteoporosis (OP) is a degenerative bone disease commonly associated with reduced bone density and increased fracture risk. Aim of the studyThis study aimed to validate the therapeutic effects of Simiao wan (SMW) on OP and explore the underlying mechanism, particularly focusing on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. Materials and methodsThe chemical components of SMW were identified using UPLC-Q-TOF-MS/MS. The obtained compounds were then input into the TCMSP, TargetNet, and SwissTargetPrediction databases to predict potential targets. OP-related targets were collected from the GeneCards and DisGeNET databases, and intersecting targets were identified through a Venn diagram. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the intersecting targets using the Database for Annotation, Visualization and Integrated Discovery (DAVID). SMW extract was subsequently used to treat osteoblasts in vitro, and its toxicity on osteoblasts was assessed using Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) release assays. Osteoblast differentiation and activity were further evaluated using alizarin red staining, alkaline phosphatase staining, and Western blot analyses to validate the activation of network pharmacological signaling pathways. ResultsA total of 121 potential targets were identified for SMW in the treatment of OP, with AKT1 as the primary target. The PI3K/AKT pathway emerged as a key signaling pathway potentially involved in SMW's therapeutic effects o OP. Toxicity assessments showed no significant toxicity of SMW on osteoblasts. Additionally, SMW promoted osteoblast proliferation, alkaline phosphatase activity, calcium nodule deposition, and the expression of osteogenic markers (osteocalcin (OCN), runt-related transcription factor 2 (RunX2), and collagen I), and activated the PI3K/AKT signaling pathway. The PI3K/AKT pathway inhibitor LY294002 partially reversed the SMW-induced mineral deposition and expression of OCN, RunX2, and collagen I. ConclusionSMW demonstrated effective multi-target and multi-pathway therapeutic potential in the treatment of OP, with a significant impact on the PI3K/AKT signaling pathway.

Tensile stress induced osteogenesis of periodontal ligament cells via Piezo1 mediated TAZ-Cbfα1 signaling

ObjectiveCyclic tensile stress (CTS) is known to induce osteogenesis of periodontal ligament cells (PDLCs), in which the molecular mechanism remains to be elucidated.This study aimed to investigate the role of the mechanosensitive calcium channel Piezo1 in the osteogenesis of PDLCs under tensile strain, as well as the signal regulation of the TAZ-Cbfα1 pathway in this process. DesignPDLCs were isolated from periodontal ligament tissues and subjected to CTS. Alizarin red staining (ARS) and alkaline phosphatase (ALP) assay were used to detect the osteogenesis of PDLCs. RT-qPCR and Western blot were used to detect the transcripts and protein expression levels of Piezo1, Transcriptional co-activator with PDZ binding motif (TAZ), and Core-binding factor α1 (Cbfα1) respectively. Immunofluorescence staining was used to detect the nuclear aggregation of TAZ. Small interfering RNA (siRNA) targeting Piezo1 (Piezo1-siRNA) was adopted to inhibit Piezo1 mRNA expression. ResultsThe results showed that the osteogenic differentiation capacity of PDLCs was significantly enhanced under CTS, along with elevated mRNA and protein expression levels of Piezo1, TAZ, and Cbfα1. Moreover, the ALP activity and the formation of calcium nodules by ARS staining were significantly increased. In addition, Piezo1 siRNA infection significantly inhibited the CTS-induced TAZ-Cbfα1 pathway and the osteogenesis of PDLCs, suggesting the regulatory role of Piezo1. ConclusionsWe provided evidence that the application of CTS encourages the osteogenic differentiation of PDLCs, which could be mediated by the Piezo1 targeted TAZ-Cbfα1 signaling.

Catalpol Enhances Osteogenic Differentiation of Human Periodontal Stem Cells and Modulates Periodontal Tissue Remodeling in an Orthodontic Tooth Movement Rat Model.

This study examines the effects and mechanisms of catalpol (CAT) on the proliferation and osteogenic differentiation of cultured human periodontal ligament stem cells (hPDLSCs) in vitro and assesses the impact of CAT on periodontal remodeling in vivo using an orthodontic tooth movement (OTM) model in rats. hPDLSCs were cultured in a laboratory setting, and their proliferation and osteogenic differentiation were assessed using the Cell-counting Kit-8 (CCK-8), Alizarin Red Staining (ARS), quantitative calcium assay, alkaline phosphatase (ALP) staining and activity assay, and immunofluorescence assay. Additionally, the expression of collagen type 1 (COL-1), ALP, and runt-related transcription factor-2 (RUNX-2) was evaluated through qRT-PCR and Western blot analysis. To verify the function of the estrogen receptor-α (ER-α)-mediated phosphatidylinositol-3-kinase-protein kinase B (PI3K/AKT) pathway in this mechanism, LY294002 (a PI3K signaling pathway inhibitor) and the ER-α specific inhibitor methyl-piperidine-pyrazole (MPP) were used. The osteogenic markers ER-α, AKT, and p-AKT (phosphoprotein kinase B) were identified through Western blot analysis. Eighteen male Sprague-Dawley rats were assigned to two groups randomly: a CAT group receiving CAT and a control group receiving an equivalent volume of saline. Micro-computed tomography (micro-CT) analysis was employed to evaluate tooth movement and changes in alveolar bone structure. Morphological changes in the periodontal tissues between the roots were investigated using hematoxylin and eosin (HE) staining and tartaric-resistant acid phosphatase (TRAP) staining. The expression of COL-1, RUNX-2, and nuclear factor-κB (NF-κB) ligand (RANKL) was assessed through immunohistochemical staining (IHC) to evaluate periodontal tissue remodeling. Tests were analyzed using GraphPad Prism 8 software. Differences among more than two groups were analyzed by one-way or two-way analysis of variance (ANOVA) followed by the Tukey's test. Values of p < 0.05 were regarded as statistically significant. In vitro experiments demonstrated that 10μM CAT significantly promoted the proliferation, ALP activity, and calcium nodule formation of hPDLSCs, with a notable increase in the expression of COL-1, ALP, RUNX-2, ER-α, and p-AKT. The PI3K/AKT pathway was inhibited by LY294002, and further analysis using MPP suggested that ER-α mediated this effect. In vivo, experiments indicated that CAT enhanced the expression of COL-1 and RUNX-2 on the tension side of rat tooth roots, reduced the number of osteoclasts on the compression side, inhibited RANKL expression, and suppressed OTM. CAT can promote hPDLSCs proliferation and osteogenic differentiation in vitro through the ER-α/PI3K/AKT pathway and enhance periodontal tissue remodeling in vivo using OTM models. These findings suggest the potential for the clinical application of catalpol in preventing relapse following OTM.

Implications of calcified nodule and adjacent calcification severity for target lesion revascularization after percutaneous coronary intervention: multi-center intravascular ultrasound imaging study

Abstract Background Calcified nodule (CN) is a distinct plaque phenotype, characterized as protruding nodular calcification penetrating lumen surface with the attached thrombus. Recent studies revealed that a risk of repeat revascularization after PCI was substantially elevated in patients with coronary artery disease (CAD) attributable to CN. Furthermore, pathohistological studies have elucidated mechanistic insights into the formulation and progression of CN, suggesting that it is formulated by the fracture of sheet calcification at its adjacent segment. Purpose To elucidate the association of calcified plaque volume at CN and adjacent lesions with TLR risks in CAD patients receiving PCI by using IVUS imaging. Methods This multicenter study analyzed IVUS imaging at culprit segment was conducted in 204 CAD patients attributable to de novo CN receiving PCI. The calc grade was assigned at each 1-mm cross-sectional frame of CN and culprit segment, respectively (no calc=0, 1-89°=1, 90-179°=2, 180-269°=3, 270-360°=4). Calcium Volume Index (CVI) at both CN and adjacent zone were calculated (Figure; Calcification volume index at CN zone = the sum of calcium scores at CN・averaged lesion slices of CN / CN lesion slices, Calcification volume index at adjacent zone = the sum of calcium scores at adjacent zone・averaged lesion slices of adjacent zone / adjacent zone slices). Predictors of TLR were analyzed by multivariate Cox hazard model. Results CN was successfully treated by DES (82%) and DCB (18%). 34% and 8.3% of subjects required rotablator and orbital atherectomy, respectively. Consequently, final lumen area after PCI was 7.2 ± 0.4 mm2. During the 2.8-year observational period (interquartile range: 2.4 to 3.2 years), TLR was required in 63 patients (30.9%) of study population. CVI at CN and adjacent zone were 9.9 ± 0.4 and 7.0 ± 0.7, respectively. On multivariate analysis, hemodialysis, final lumen area, CVI at both CN and adjacent zone predicted TLR (Figure). Recurrent TLR (two or more TLRs) was found in 23.3% patients who suffered from TLR. Of note, even among patients who required TLR, CVIs at both zones were significantly higher in patients with recurrent TLR than those without (Figure). Conclusion CAD subjects caused by CN exhibited a considerably high-risk TLRs. Our findings underscore that calcification severity at both CN and adjacent segment predicted TLR and is associated with repeated TLR.methodsResults

Significant stenosis without thrombus may be the third most common etiology of acute coronary syndrome

Abstract Background Plaque rupture (PR), plaque erosion (PE) and calcified nodule (CN) are known as major etiologies of acute coronary syndrome (ACS). On the other hand, significant stenosis without thrombus (SSWT) can be sometimes observed in culprit plaque of ACS. However, incidence and clinical feature of SSWT in ACS patients are not well known in Japanese. Method This is a sub study of TACTICS registry which include ACS patients (n=702) undergoing primary percutaneous coronary intervention (PCI) under optical coherence tomography (OCT) in Japan. Using this registry data, we compared the clinical features and OCT findings of SSWT in ACS patients in comparison with PR, PE and CN. Major adverse cardiac event (MACE) was defined as death, non-fatal myocardial infarction, heart failure, or ischemia-driven revascularization at 1-year. Results Plaque rupture (PR: n=411, 61%) and plaque erosion (PE: n=178, 26%) were 2 major etiologies, which was followed by SSWT (n=59, 9%) and calcified nodule (CN: n=28, 4%). Clinical characteristics are shown in table 1. Patients with SSWT was slightly older than PR and PE but younger than CN. Proportion of male was fewer than PR and PE but greater than CN. In terms of traditional coronary risk factors, there were significant differences in prevalences of diabetes, active smoking, and hemodialysis among different etiologies. In SSWT, non-ST elevation myocardial infarction was the main type of ACS followed by unstable angina and ST elevation myocardial infarction (51%, 39% and 10%, respectively), which may be associated with the least CK level and greater TIMI flow. Lesion was less complex with lower proportion of type B2 or C, shorter procedure time, lower proportion of multivessel disease and lower syntax score. OCT findings are shown in table 2. Regarding lipid component and calcification, SSWT showed similar finding as PE. Cholesterol crystal was relatively less as compared to PR and PE. Kaplan-Meier survival curve showing MACE and ischemia driven-TLR are shown in figure 1 and 2. MACE and TLR was the least in SSWT. Conclusion SSWT was the third most common etiology of ACS with different feature from PR, PE and CN.

Impact of sex and age on culprit plaque type of acute coronary syndrome in Japanese patients from TACTICS registry

Abstract Background In acute coronary syndrome (ACS), plaque rupture (PR) is the most frequent etiology followed by plaque erosion (PE) while calcified nodule (CN) is the least etiology. These three are known as major etiologies of ACS. Traditionally, it has been believed that sex and age have impact on etiology of ACS. For example, plaque erosion was thought to often occur in young female while calcified nodule often occur in elderly patients. However, impact of sex and age have not been well described. Methods This is a sub-study from TACTICS registry which consist of 702 Japanese ACS patients who underwent primary percutaneous coronary intervention under optical coherence tomography (OCT) from multicenter. Based on the OCT findings, patients were divided into PR, PE, CN and others. These data were stratified by sex and ages (&amp;lt;50, 50s, 60s, 70s, 80 or over). In this sub-study, we mainly focused on 3 major etiologies of ACS (PR, PE, CN) and compared incidence and clinical characteristics based on sex and ages. Results Distribution of culprit plaque type in ACS patients are shown in figure 1A (male) and 1B (female). PR remained top etiology of ACS in any sex and any age category. However, trend of PE was different between male and female. In male, as patients get younger, incidence of PE increased. In male patients under 50, incidence of PE was almost similar as PR. On the other hand, in female, incidence of PE was greatest in 70s followed by 80 or over. Surprisingly, young female showed the least incidence of PE. CN was the least etiology of three but increased especially from 80. Clinical characteristics are shown in table 1A (male) and 1B (female). In male, most of the baseline characteristics differ between different ages while in female, most of the baseline characteristic did not differ between different ages. Conclusion In our data, PE was more distinct in younger male ACS patients while PE was not that distinct in younger female ACS patients. The impact of sex and age on culprit plaque type of ACS might be variable depending on population.

Coronary artery plaque characteristics in smokers assessed by intracoronary multi-modality imaging: a NIRS-IVUS guided PROSPECT II substudy

Abstract Background Smoking is a risk factor for coronary artery disease and myocardial infarction. Purpose Among patients with myocardial infarction, we hypothesized smokers would have increased prevalence of nonculprit high-risk plaque characteristics as assessed by intracoronary multimodality imaged when compared to nonsmokers. Method This is a prespecified post-hoc analysis of the PROSPECT II (Providing Regional Observations to Study Predictors of Events in the Coronary Tree). Altogether 898 patients with acute myocardial infarction with or without ST-segment elevation who after successful percutaneous coronary intervention had near-infrared spectroscopy and intravascular ultrasound imaging performed. Data on lesion location, plaque burden, and lipid content were collected. Current, former, and nonsmokers were examined regarding the presence of high-risk plaques in both culprit and nonculprit lesions as well as plaque location. High-risk plaque characteristics were defined as a maximum plaque burden (maxPB) of ≥70% and a maximum 4 mm lipid core burden index (maxLCBI4mm) of ≥324.7. Results Out of 886 patients with known smoking history, 32% were smokers, 32% former, and 37% nonsmokers. Smokers were younger (median age: smokers 59.0; former 65.5; nonsmokers 64.0 years, p&amp;lt;0.001) with fewer co-morbidities and presented more often with STEMI (% presenting with STEMI: smokers 22.2%; former 18.9%; nonsmokers 19.4%, p=0.006). LDL was similar but CRP was higher in smokers (median µg/mL: smokers 4.0; former 3.1; nonsmokers 2.9, p=0.004). In multivariate models of nonculprit lesion morphology smoking history was not associated with maxPB or maxLCBI4mm but was associated with presence of calcified nodules (OR: 2.88 (95% CI 1.46–5.69) p=0.002 [smokers vs nonsmokers]). Calcified nodules were also associated with number of years smoked (OR: 1.02 (95% CI 1.01–1.03) p=0.004). Smoking was not associated with presence of ≥1 lesions with MaxLCBI4mm, ≥ 324.7, maxPB &amp;gt;=70% or the combination of both. Smoking was not associated with presence of plaque rupture or thrombus. Right coronary artery culprit lesions were more common in smokers and former smokers (smokers 29.9%; former 30.9%; nonsmokers 21.6%; OR: 1.12 (95% CI 1.04–1.21) p&amp;lt;0.001 [smokers vs nonsmokers]). Conclusion In this prospective intracoronary multimodality imaging study of patients with recent acute myocardial infarction, smoking was not associated with a higher rate of high-risk nonculprit plaques based on lipid content by NIRS or PB by IVUS when compared to former or nonsmokers. However, smoking was associated with higher presence of IVUS-detected calcified nodules, higher CRP-levels, and the right coronary artery as it angiographically was a more common site for culprit lesions. These results indicate that the elevated risk in smokers is unrelated to vulnerable plaque morphology defined by high PB and lipid content, and instead possibly related to inflammation, calcification, or hemodynamic changes.Table 1

The impact of SYNTAX score on culprit lesion morphology of acute coronary syndrome

Abstract Background The SYNTAX score quantifies the complexity of coronary artery morphology and is useful in determining the indication for CABG (Coronary Artery Bypass Grafting) or PCI (Percutaneous Coronary Intervention), which often reflect coronary artery disease severity. However, how SYNTAX score affects etiology of ACS (Acute Coronary Syndrome) patients observed by OCT (Optical Coherence Tomography) has been unclear. Methods This is a sub study of TACTICS OCT registry (n=702) which include ACS patients who underwent OCT guided PCI. Based on SYNTAX score, patients were divided into low-SYNTAX-group (1-22 points) and high-SYNTAX-group (23 points or higher). ACS culprit lesions were classified into plaque rupture (PR), plaque erosion (PE), and calcified nodule (CN). We compared patient characteristics and culprit lesions in low-group and high-group. Results High-SYNTAX-group was significantly younger with relatively greater prevalence of diabetes and lower body mass index. Peak CK, LVEF, Multi vessel and TIMI grade were also significantly greater in high-SYNTAX-group than low-SYNTAX-group. Although PR and PE were 2 most common etiologies of ACS in both groups, proportion of CN was much greater in high-SYNTAX-group than low-SYNTAX-group. Conclusion High-SYNTAX score was likely to have impact on ACS etiology increasing proportion of CN.

Comparison of different calcium modification strategies for coronary calcified nodules: insights from optical coherence tomography

Abstract Background The optimal treatment strategy for coronary calcified nodules (CN) remains uncertain. Purpose We sought to compare the safety and efficacy of different calcium modification strategies for managing CN using optical coherence tomography (OCT). Methods In this single-center retrospective cohort study, consecutive patients undergoing OCT-guided percutaneous coronary intervention (PCI) for severely calcified lesions using rotational atherectomy (RA), orbital atherectomy (OA), or intravascular lithotripsy (IVL) between January 2017 and December 2022 were included. OCT analysis was performed through our institutional core-laboratory to identify CN, assess plaque characteristics, and evaluate post-stent optimization. Primary outcomes were minimum stent area (MSA) and MSA at CN site post-PCI. One-year target vessel failure (TVF), defined as a composite of cardiac death, target-vessel myocardial infarction, or ischemia-driven target vessel revascularization, was also recorded. Results Among a total of 154 patients with 158 lesions, CN was identified in 54 lesions (34.2%) and managed with RA (39%, n=21), OA (33%, n=18), or IVL (28%, n=15). The IVL group exhibited a larger minimal lumen diameter, maximum calcium arc, and maximum calcium thickness at baseline. Post-PCI OCT demonstrated comparable MSA (RA: 5.91 ± 0.33mm², OA: 5.94 ± 0.38mm², IVL: 6.46 ± 0.45mm²; p=0.61) and MSA at CN site (6.77 ± 0.49mm², 6.71 ± 0.51mm², 7.13 ± 0.59mm², respectively; p=0.87) after adjusting for procedural and morphologic factors among the groups. The incidence of TVF at one year was similar among the groups. Conclusions In patients undergoing PCI for severely calcified lesions with CN, comparable procedural and clinical outcomes can be achieved using RA, OA, or IVL. The equivalence across these techniques underscores a role for individualized decision-making integrating patient- and lesion-specific characteristics, as well as center expertise when selecting a treatment strategy for CN.OCT findings and clinical outcomes