micro-CT Analysis Research Articles

Preheated and Injected Bulk-Fill Resin Composites: A Micro-CT Analysis of Internal Voids and Marginal Adaptation in Class II Restorations

The aim of this study was to evaluate, in vitro, the void formation and marginal adaptation in Class II cavities restored with preheated and injected bulk-fill resin composites. Eighty third molars received Class II cavities on their mesial and distal surfaces and were randomly distributed into eight groups (n = 10) according to material (Filtek Universal—control, incremental technique; Filtek One Bulk-Fill; Admira Fusion X-tra Bulk-Fill; VisCalor Bulk-Fill) and the temperature of the material (24 °C or 68 °C). The restored teeth were scanned using a SkyScan 1173 microtomograph. The percentage of internal voids (%IV) was analyzed using CTan software (version 1.23.02) and the percentages of continuous margins (%CM) in enamel and dentin were analyzed using Dataviewer software (version 1.5.6.2). The data of %IV and %CM were subjected to two-way ANOVA on ranks, followed by Tukey’s test (α = 0.05). At 24 °C, Filtek Universal had a greater %IV (1.89%) (p < 0.05), which did not differ significantly from that of Admira Fusion X-tra Bulk-Fill (0.29%) (p > 0.05). Filtek One Bulk-Fill (0.07%) and VisCalor Bulk-Fill (0.07%) had lower %IV (p < 0.05). Preheating resulted in a significantly lower %IV for Admira Fusion X-tra Bulk-Fill (p < 0.05). Temperature did not significantly influence marginal adaptation (p > 0.05). VisCalor Bulk-Fill achieved significantly higher %CM in dentin (98%) at 24 °C (p < 0.05). It was concluded that bulk-fill-injected resin composites tend to have fewer internal voids than conventional resin composites using the incremental technique, and the injection of the resin composite into the cavity seems to be more important for marginal adaptation than the preheating procedure.

Sinomenine attenuates uremia vascular calcification by miR-143-5p

Vascular calcification is considered to be a killer of the cardiovascular system, involved inflammation and immunity. There is no approved therapeutic strategy for the prevention of vascular calcification. Sinomenine exhibited anti-inflammatory and immunosuppressive effects. Objective of this study was to investigate the effect of sinomenine in vascular calcification and its potential molecular mechanism. Adenine-induced uremic rats were constructed and administrated with sinomenine. Optical clearing of aortas, alizarin red staining, von Kossa staining, calcification quantification, micro-CT analyses of vascular calcification were performed to analyze calcification in aortas. Administration of 40 mg/kg/d sinomenine effectively alleviated vascular calcification in uremic rats. The miRNA sequencing revealed differentially expressed miRNAs in aortas and bioinformatic analysis assisted with miRNA screening. We screened 9 differential expressed miRNAs and their predicted target genes. By qRT-PCR, we validated that the expression of rno-miR-143-5p was corresponding to our prediction. Sinomenine inhibited vascular smooth muscle cells (VSMCs) calcification, accompanied with miR-143-5p upregulation. MiR-143-5p mimic decreased VSMCs calcification in high phosphate condition. On the contrary, miR-143-5p inhibitor increased VSMCs calcification in high phosphate condition, which was inhibited by sinomenine. In chronic kidney disease patients with vascular calcification, the expression level of circulating miR-143-5p was lower than those without vascular calcification. Sinomenine significantly inhibited vascular calcification in VSMCs and uremic rat. MiR-143-5p was one of the collection of miRNAs modified by sinomenine in vascular calcification. Reduction of miR-143-5p in VSMCs was not only a concomitant phenomenon in pro-calcification condition but also contribute to VSMCs calcification. Circulating miR-143-5p was supposed to be a potential biomarker for vascular calcification in chronic kidney disease patients. In conclusion, sinomenine effectively alleviated vascular calcification, which was attributed to miR-143-5p regulation partly.

The microRNA-455 null mouse shows dysregulated bone turnover

Abstract A wide range of specific microRNAs have been shown to have either positive or negative effects on osteoblast differentiation and function with consequent changes in post-natal bone mass; a number of specific targets have been identified. We previously used CrispR-Cas9 to make a miR-455 null mouse, characterising a behavioural phenotype with age. The current study identifies a bone phenotype, starting in younger animals. At three weeks of age, the miR-455 null mice (both male and female) display increased length of both long bones and vertebrae and whilst this difference diminishes across 1 year, it remains significant. Increased bone formation in vivo is mirrored by an increase in osteogenesis from bone marrow-derived stem cells in vitro. This is accompanied by a decrease in osteoclastogenesis and osteoclast function. MicroCT analyses show increased trabecular bone and less porosity/decreased separation in the miR-455 null mouse suggesting a more dense and stronger bone at three weeks of age, these differences normalise by 1 year. Gain-of-function and loss-of-function datasets shows that FGF18 expression is regulated by miR-455, and FGF18 was validated as a direct target of miR-455. The regulation of FGF18 by miR-455 is a likely mediator of its effect on bone.

Evaluation of a potential bidirectional influence of metabolic syndrome and apical periodontitis: An animal-based study.

This study aimed to explore the possible bidirectional interrelations between fructose-induced metabolic syndrome (MS) and apical periodontitis (AP). Twenty-eight male Wistar rats were distributed into four groups (n = 7, per group): Control (C), AP, Fructose Consumption (FRUT) and Fructose Consumption and AP (FRUT+AP). The rats in groups C and AP received filtered water, while those in groups FRUT and FRUT+AP received a 20% fructose solution mixed with water to induce MS. The groups AP and FRUT+AP had the pulp of their right mandibular first molar exposed to induce AP. Food consumption, murinometric measurements, blood glucose levels and glucose tolerance were monitored. Fifty-six days after the start of the experiment, the animals were euthanized, and serum samples were collected for metabolomic analysis. Mandibles, livers and right kidneys were also collected. The area and volume of the periapical lesions were calculated using micro-computed tomography. Histopathological evaluation was performed. Kruskal-Wallis followed by the Student-Newman-Keuls or Mann-Whitney tests and one-way anova followed by Tukey's or Independent t-test were used for non-parametric and parametric data, respectively (p < .05). Multivariate analysis and variable importance in projection score were applied to assess metabolite profile differences among groups (p < .05). FRUT and FRUT+AP groups showed significantly increased fluid intake, body mass, abdominal circumference, blood glucose levels, liver weight and visceral fat weight (p < .05), indicating the development of MS. The analyses of the metabolite profile suggest increasing glucose, histidine, lactate, fatty acid and phenylalanine in the FRUT+AP group. There were no significant differences in volume and area of periapical lesions in micro-CT analyses (p = .1048 and p = .7494, respectively). Histopathological analysis of the hemimandibles demonstrated areas of inflammatory response, necrosis and microabscess in the periapical region. Hepatic histopathological observations indicated notable differences in cell appearance, with the FRUT and FRUT+AP groups showing signs of microsteatosis. Kidney analysis revealed Bowman's space dilation in the FRUT and AP groups, while the FRUT+AP group exhibited retracted Bowman's space, suggesting a possible alteration in renal filtration capacity. MS had no impact on the progression of AP in rats. However, AP exacerbated the systemic state affected by MS, with changes in liver and kidney tissues and metabolite levels.

Fraxin inhibits ovariectomized-induced bone loss and osteoclastogenesis by suppressing ROS activity.

Osteoporosis is characterized by increased osteoclast activity, which is strongly associated with increased levels of reactive oxygen species (ROS). Fraxin, a natural coumarin glycoside, has shown anti-inflammatory and antioxidant properties, but its effects on bone homeostasis are obscure. The effects of fraxin on osteoclast formation and activation were measured via an in vitro osteoclastogenesis assay. Mitochondrial and total ROS production were evaluated with the aid of MitoSOX Red and DCFH-DA, respectively. Osteoclast-related gene expression analysis was performed via qPCR. Key proteins related to osteoclast formation, ROS scavenging, and ROS-regulated signaling, such as mitogen-activated protein kinases (MAPKs), NF-κB pathways, and nuclear factor of activated T cells 1 (NFATc1) signaling, were detected via western blotting. An ovariectomized mouse model was used to evaluate the therapeutic effects of fraxin in vivo. Fraxin inhibited osteoclastogenesis and osteoclast-related gene expression. Mechanistically, fraxin restored the levels of ROS-scavenging enzymes to inhibit ROS accumulation, eventually downregulating ROS-regulated signaling. The measurement of Micro-CT and histological analyses revealed that fraxin treatment significantly reduced OVX-induced bone loss by decreasing the number of osteoclasts. Fraxin shows promise as a novel therapeutic agent for osteoclast-related bone diseases, especially osteoporosis.

Abaloparatide Enhances Bone Regeneration in Extraction Socket Dental Implant Defects: An Experimental InVivo Study.

Abaloparatide (ABL) is a synthetic parathyroid hormone-related protein analog developed as an anabolic drug to treat osteoporosis. ABL increases bone mineral density (BMD) of the long bones and spine; however, the influence of ABL on alveolar bone regeneration remains unknown. This study assessed the effects of systemic ABL administration on tooth extraction socket healing and dental implant osseointegration in a preclinical rodent model. Sprague-Dawley rats received daily subcutaneous injection of ABL (25 μg/kg) or vehicle (VEH) at the time of maxillary first molar (M1) extraction surgery; animals underwent either: (1) unilateral M1 extraction followed by sacrifice at 10 or 42d or (2)bilateral M1 extraction followed by staged implant placement in osteotomies with standardized defects and sacrifice at 21or 28d. Micro-computed tomography (micro-CT) analysis demonstrated that ABL increased bone volume fraction (p = 0.004) and bone mineral density (BMD) (p = 0.006) of regenerated extraction sockets at 42d. Micro-CT of the femur validated systemic effects of ABL, showing greater trabecular BMD after 6 (p < 0.01), 9 (p < 0.001), and 14 (p < 0.001) weeks of treatment. Histomorphometry confirmed a higher mineralized bone area with ABL treatment in extraction sockets at 42d (p = 0.03) and in the regenerated peri-implant bone at 21d post-implant placement (p = 0.01) compared to control. Systemic ABL administration enhances osteogenesis in extraction sockets prior to implant placement and accelerates peri-implant bone formation in the early phase of healing in the present rodent model.

Forsythia viridissima Leaf Extract Inhibits Trabecular Bone Mineral Loss in Rats Induced by a Low-Calcium Diet.

The Forsythia has been used in herbal medicine, and the leaf is also expected to contain various putative bioactive substances. In this study, we investigated the effects of Forsythia viridissima leaf extract (FLE) on bone metabolism. The anti-osteoporotic effect of FLE was determined in male rats fed a low-calcium diet. Micro-CT analysis of the distal end of the left femur showed a trend toward less reduction in total bone mineral density (total BMD) in the group received FLE on a 0.05% low-calcium diet compared to the low-calcium control group. Also, trabecular bone mineral density (Tb.BMD) was significantly less decreased in the FLE group. Then, in vitro experiment, we found that matairesinol, a lignan particularly rich in Forsythia leaf, remarkably promotes osteoblastic mineralization. Thus, our study revealed that FLE inhibits the progression of osteoporosis caused by calcium deficiency.

Puerarin Promotes Tibial Shaft Fracture Healing in Rats and Its Relationship with BMP-Smad Pathway

Background Fracture healing involves multiple growth factors and the interaction between multiple signaling pathways. Pue (puerarin) is a natural compound with multiple biological activities, but its effect on fracture healing and its relationship with the bone morphogenic protein (BMP)-Smad pathway is not yet clear. Purpose This study intends to assess the effect of puerarin on fracture healing. Methods Seventy rats were used for the study to establish a tibial shaft fracture rat model. The experimental rats were divided into a tibial fracture group, Pue dose group (low, medium, and high), BMP-2 group, Noggin group, Pue + SY-LB-35 group, and Pue + Noggin group through the random number method. We observed the fracture healing through micro-CT and histology, observed the tibial stump tissue with hematoxylin and eosin (HE) staining, and detected the expression of genes related to the BMP-Smad pathway through quantitative real-time polymerase chain reaction (qPCR) and Western blot. Results We found that puerarin can promote tibial shaft fracture healing in rats in a dose-dependent manner; through micro-CT and histological analysis, we observed increased trabecular bone formation during fracture healing in rats treated with puerarin. Callus formation is accelerated, and bone density increases. In addition, puerarin also significantly increased the expression of genes related to the BMP-Smad pathway, including BMP2, Smad1, Smad5, and so on. Conclusion Puerarin can promote tibial shaft fracture healing in rats, and its mechanism is related to upregulating the expression of genes related to the BMP-Smad pathway. It further improves the fracture-healing mechanism and provides candidate drugs for the treatment of tibial shaft fracture healing in clinical medicine.

Obturation quality of bioceramic sealers with different obturation techniques: a micro-CT evaluation

This micro-computed tomography (micro-CT) analysis in vitro study was designed to compare void volume in root canal fillings performed using the single-cone (SC) technique and the continuous wave condensation (CWC) technique with bioceramic (BC) sealer. Forty human-extracted, single-rooted mandibular premolars were cleaned, shaped, and divided into two groups (n = 20) based on the obturation technique. In the first group, obturation was performed using the CWC technique with TotalFill HiFlow BC sealer. In the second group, obturation was performed using the SC technique with the conventional TotalFill BC sealer. All roots were scanned with micro-CT, and ©CTAn software version 1.20.8.0 was used to measure the void volume of each root third in all samples. Despite the different obturation techniques, both groups exhibited voids in filled root canals. The overall mean void volume in the SC group (0.253 ± 0.186) was significantly higher (p = 0.000) than that in the CWC group (0.035 ± 0.043). The difference was significant in the coronal (p = 0.000) and middle (p = 0.017) thirds. The CWC technique with TotalFill HiFlow BC sealer achieved superior obturation quality compared to the SC technique with conventional BC sealer in oval-shaped premolars.

The Application of Resolvin D1-Loaded Gelatin Methacrylate in a Rat Periodontitis Model

Objective: To evaluate the drug release, cytocompatibility with periodontal ligament cells (PDLCs), and therapeutic efficacy of GelMA hydrogel loaded with resolvin D1 (RvD1) in treating rat periodontal inflammation and alveolar bone damage. Methods: An RvD1 complexed with GelMA was prepared, and its release kinetics and compatibility with PDLCs were assessed. Rats with induced periodontitis were treated weekly with topical applications of vehicle, GelMA, RvD1, or RvD1 complexed with GelMA for four weeks. Periodontal inflammation and tissue regeneration were evaluated using quantitative PCR (qPCR) and histochemical staining, while alveolar bone repair and regeneration were analyzed through micro-CT. Results: The RvD1 complexed with GelMA effectively released RvD1 and enhanced the proliferation and viability of PDLCs. Compared to RvD1 alone, treatment with RvD1 complexed with GelMA significantly reduced inflammatory cell infiltration, TNF-α and RANKL expression, and osteoclast formation in periodontal tissues. Additionally, it promoted the expression of specific anti-inflammatory and regenerative markers. Micro-CT analysis confirmed significant new bone formation in the RvD1 complexed with GelMA-treated group. Conclusions: RvD1 complexed with GelMA provides sustained drug release and biocompatibility, effectively resolves periodontal inflammation, and promotes tissue regeneration in periodontitis.

Microstructural Analysis of the Human Scapula: Mandibular Bone Tissue Engineering Perspectives.

Mandibular bone defect reconstruction remains a significant challenge for surgeons worldwide. Among multiple biodegradable biopolymers, allogeneic bone scaffolds derived from human sources have been used as an alternative to autologous bone grafts, providing optimal conditions for cell recruitment, adhesion, and proliferation and demonstrating significant osteogenic properties. This study aims to investigate the bone microstructure of the human scapula as a source for allogeneic bone scaffold fabrication for mandibular tissue engineering purposes. We created color-coded anatomical maps of the scapula and the mandible, reflecting the best anatomical and geometrical match. In this pilot study, we hypothesized a microstructural similarity of these bone structures and evaluated the human scapula's bone tissue engineering potential for mandibular bone tissue engineering by focusing on the microstructural characteristics. Lyophilized human scapular and mandibular bioimplants were manufactured and sterilized. Experimental bone samples from the scapula's acromion, coracoid, and lateral border from the mandibular condyle, mandibular angle, and mental protuberance were harvested and analyzed using micro-CT and quantitative morphometric analysis. This pilot study demonstrates significant microstructural qualitative and quantitative intra-group differences in the scapular and mandibular experimental bone samples harvested from the various anatomical regions. The revealed microstructural similarity of the human scapular and mandibular bone samples, to a certain extent, supports the stated hypothesis and, thus, allows us to suggest the human scapula as an alternative off-the-shelf allogeneic scaffold for mandibular reconstruction and bone tissue engineering applications.

Genetic Ablation of p16 Mitigates Premature Osteoporosis Induced by PTHrP Nuclear Localization Sequence and C-terminal Deletion through Inhibition of Cellular Senescence.

Premature osteoporosis due to parathyroid hormone-related peptide (PTHrP) dysfunction presents significant bone health challenges. This study explores the role of p16-mediated cellular senescence in this condition using a Pthrp knock-in (KI) mouse model lacking the nuclear localization sequence and C-terminus of PTHrP. We generated p16⁻⁄⁻KI mice and compared them with wild-type, p16⁻⁄⁻, and KI mice. The genetic ablation of p16 in KI mice extended their lifespan, increased body size, and weight. Micro-CT analysis revealed a significant increase in bone volume, while histological and immunohistochemical studies revealed enhanced chondrocyte proliferation and osteoblast function in p16⁻⁄⁻KI mice. In vitro experiments showed enhanced differentiation capacity and reduced senescence of bone marrow mesenchymal stem cells (BM-MSCs) from p16⁻⁄⁻KI mice. Molecular analyses indicated that p16 knockout partially reversed oxidative stress, DNA damage, and cellular senescence observed in KI mice, as evidenced by upregulated antioxidant enzymes, reduced DNA damage markers, and decreased senescence markers. These findings highlight the critical role of p16-mediated cellular senescence in the premature osteoporosis phenotype of KI mice, suggesting that targeting cellular senescence pathways could offer a promising therapeutic strategy for premature osteoporosis and age-related bone loss. This research provides new insights into the interplay between genetic factors, cellular senescence, and bone metabolism in the context of aging and osteoporosis.

Delayed Tooth Development and the Impaired Differentiation of Stem/Progenitor Cells in Incisors from Type 2 Diabetes Mice.

Patients with diabetes mellitus (DM) have an increased risk of tooth decay caused by alterations in their tooth development and their oral environment, as well as a tendency to present with pulp infection due to compromised immune response. The present study analyzed the characteristic alterations in tooth development under DM conditions using incisors from db/db type 2 diabetic mouse model (T2DM mice). In micro-CT analyses, T2DM mice showed delayed dentin and enamel formation. Through transcriptomic analyses, pre-ameloblast- and pre-odontoblast-specific genes were found to be significantly decreased in the incisors of T2DM mice, whereas major ameloblast- and mature odontoblast-specific genes were not changed. Stem cell markers were decreased in T2DM mice compared to those from the control mice, suggesting that the stemness of dental pulp cells (DPCs) is attenuated in T2DM mice. In vitro analyses demonstrated that DPCs from T2DM mice have lower colony-forming units (CFU), slower propagation, and diminished differentiation characteristics compared to the control. These data suggest that T2DM conditions could impair the differentiation property of multiple progenitor/stem cells in the tooth, resulting in delayed tooth development in T2DM mice.

Proteomic and Morphological Analysis of Bone and Articular Cartilage Changes in Osteoarthritic Rabbits Supplemented with Edible Bird’s Nest (EBN)

This study is aimed at assessing the effects of Edible Bird’s Nest (EBN) on subchondral bone, articular cartilage and the expression of proteins in synovial fluid by micro-CT evaluation and histological analysis. 54 New Zealand white rabbits were induced by intra-articular injection of monosodium iodoacetate (8 mg) and divided into four groups: negative control (n=9): non-treated osteoarthritis; positive control (n=15): OA + diclofenac sodium 2 mg/kg daily orally; low dosage (n=15): OA + 75 mg/kg hydrolyzed EBN; and high dosage (n=15): OA + 150 mg/kg hydrolyzed EBN. The joints were harvested and subjected to micro-CT analysis and histological evaluation, and the synovial fluid was subjected to LCMS/MS analysis. Micro-CT analysis showed an increase in bone volume and a decrease in total porosity in the treatment group that showed bone integrity improvement. Histopathological results revealed comparable changes between the positive control group and the EBN treatment group. There was upregulation of proteins involved in the resolution of inflammation and downregulation of proteins associated with the bone resorption process. Morphology evaluation showed that EBN supplementation has a bone-improving effect by inhibiting osteoclastic activity. Protein expression showed chondroprotection and bone improvement through the action of several proteins via various signaling pathways. The morphological and molecular findings suggest the potential use of EBN as beneficial alternative for osteoarthritis treatment by improving bone quality and modulating inflammatory responses.

Optimisation of process parameters in Arburg Plastic Freeforming for enhanced part density and geometric accuracy

Technological advances have increased the use of plastic-based additive manufacturing for production in a variety of industries that require high mechanical properties while maintaining geometric precision. The Arburg Plastic Freeformer (APF) process, unlike other filament-based technologies, uses standard plastic granules used in mass production in injection moulding machines. This study focuses on optimising the interaction of three critical process parameters: layer thickness (LT), droplet aspect ratio (DAR) and discharge rate (DR). Previous studies have mainly varied individual parameters to evaluate mechanical and geometrical properties. In contrast, this work analyses these parameters as a whole and evaluates their combined effects on residual porosity and geometric accuracy. APF relies heavily on supports to sustain the printed part, with a smaller achievable self-supporting overhang angle compared to the Fused Filament Fabrication (FFF) printing process. For this reason, this study investigates two compatible materials, ABS Terluran GP35 and a water-soluble PVP compound named Armat11. Cylindrical and cubic samples were 3D printed using different combinations of LT, DAR and DR. A total of 150 cubic samples were printed to assess the geometric dimensional accuracy and repeatability of printing in the plate position using permutations in the printing plate. Subsequently, 30 cylindrical samples were printed for micro-CT analysis, reconstructed by CT file segmentation and compared with the ideal CAD model, in addition to the characterisation of residual porosity. The results show that optimal combinations of LT, DAR and DR produce high density parts with repeatable geometric properties. In addition, a quantitative analytical model was developed to optimise arbitrary parameter sets. This comprehensive investigation provides important insights into the APF process and increases its potential for wider industrial applications.

Histological Evaluation of Sodium Iodide-Based Root Canal Filling Materials in Canine Teeth.

A novel water-soluble root canal filling material based on sodium iodide (NaI) has been developed to overcome the limitations of existing iodine-based formulations. However, the biological stability of this approach in animal studies remains unverified. This study evaluated the biocompatibility of NaI compared to commercial root canal filling materials (Calcipex II and Vitapex®) in pulpectomized canine teeth to assess its clinical applicability. Following a four-week observation period, none of the experimental groups exhibited tooth mobility or fistula formation. Radiographic and micro-CT analyses revealed no radiolucency in periapical lesions. Histopathologic evaluation demonstrated the absence of inflammatory responses in periapical regions across all material groups, with histological inflammation scoring 0. High-magnification histological examination of periapical areas showed well-preserved periodontal ligament tissue in all groups. Despite certain limitations of NaI-based fillings in the pulp cavity, including loss of radiopacity and tooth discoloration, NaI demonstrates potential as a safe and effective alternative for pulp filling material, particularly due to its minimal risk of root resorption and inflammatory response.

Effect of Decellularized Amniotic Membrane on the Tendon-Bone Integration in Rotator Cuff Repair: A Comparative Rat Model Study.

Rotator cuff retear after arthroscopy repair is a difficult complication that is often due to poor tendon-bone healing. Decellularized amniotic membrane (DAM) has a variety of bioactive substances which have great potential to enhance tendon-bone healing. However, DAM has three layers, of which the middle basement layer is dense and thick. Whether DAM will hinder tendon-bone healing of rotator cuff after surgical repair is unclear. Our study aims to investigate the effect of DAM on tendon-bone healing of the rotator cuff after surgical repair. Thirty-three Sprague-Dawley (SD) rats were selected to establish unilateral supraspinatus (ST) tear models and were randomly treated with only suturing repair (OSR group, n = 11), and suturing repair with DAM placed between the ST and bone (DAM group, n = 11). In the normal control group (NCT group, n = 11), the supraspinatus was only exposed but not detached or repaired. After 4 weeks the rats were sacrificed. The assessment of specimens was conducted by micro-CT analysis, histopathological evaluation, and biomechanical testing. The DAM group had a significantly higher ultimate load to failure, new bone volume, and histological evaluation at 4 weeks after surgery than the OSR group. When comparing the DAM group to the NCT group, the DAM group performed slightly worse in biomechanical testing, micro-CT analysis, and histological evaluation. When placed between tendon and bone at the rotator cuff footprint, DAM, despite its dense and thick basement layer, does not impede tendon-bone healing after surgical repair for rotator cuff injury, but rather promotes increased healing quality and biomechanical properties. However, the healing quality and biomechanical properties are still lower than that of the normal rotator cuff, and further improvement should be made to the application strategy of a DAM.

Platelet-rich fibrin associated to bovine bone induces bone regeneration in model of critical-sized calvaria defect of rats submitted to Zoledronic Acid therapy: PRF induces bone healing

Reconstruction of bone defects prior to implant installation is a challenge, especially when the patient uses bisphosphonates. Given this difficulty, many studies investigate biomaterials that can improve the bone regeneration process. In this context, this study aimed to investigate the effect of platelet-rich fibrin (PRF) and Bio-Oss (BO) on bone regeneration of rats submitted to critical-sized calvaria defects and treated with ZA. Thirty Wistar rats received a single dose of ZA (120 μg/kg) and after 7 days, were submitted to an 8 mm calvaria defect. The animals were divided into 5 groups (n=6): ZA, BO, PRF or BO+PRF; animals from control group did not receive ZA. All animals were euthanized 12 weeks after surgical procedure and calvaria collected to histological, histomorphometric and micro-CT analyses. BO+PRF increased the number of osteoblasts (33%) and osteoclasts (58%), as well as blood vessels (70%) and Type I collagen (52%) (p<0.05) compared to ZA group. In summary, the association of BO+PRF improved bone healing of large bone defect in rats receiving ZA and this may be an interesting approach for the treatment to be tested in patients under anti-resorptive therapy.

Unveiling the Therapeutic Potential of Berberine in Rheumatoid Arthritis: A Comprehensive Study of Network Pharmacology, Metabolomics, and Intestinal Flora.

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease influenced by environmental triggers, including the commensal microbiota. Recent research has highlighted distinctive features of the gut microbiota in RA patients. This study investigates the therapeutic potential of berberine (BBR), a gut microbiota modulator known for its significant anti-RA effects, and elucidates the underlying mechanisms. Utilizing the collagen-induced arthritis (CIA) rat model, we comprehensively evaluated the anti-rheumatoid arthritis effects of BBR in vivo through various indices, such as paw edema, arthritis index, ankle diameter, inflammatory cytokine levels, pathological conditions, and micro-CT analysis. Employing network pharmacology, we identified potential targets involved in RA alleviation by BBR. To analyze comprehensive metabolic profiles and identify underlying metabolic pathways, we conducted a serum-based widely targeted metabolomics analysis utilizing LC-MS technology. An integrated network encompassing metabolomics and network pharmacology data was constructed using Cytoscape. The potential therapeutic targets and signaling pathways of BBR in the management of RA were predicted using network pharmacology. Key targets and pathways were further validated by molecular docking and immunofluorescent staining, which integrated findings from serum metabolomics and network pharmacology analysis. Additionally, we analyzed the gut microbiota composition in rats employing 16S rDNA sequencing and investigated the effects of BBR on the microbiota of CIA rats through bioinformatics and statistical methods. Our results showed that BBR demonstrated significant efficacy in alleviating RA symptoms in CIA rats, as evidenced by improvements in paw redness and swelling, attenuation of bone and cartilage damage, reduction in synovial hyperplasia, inflammatory cell infiltration, and suppression of proinflammatory cytokines IL-1β, IL-6, IL-17A, and TNF-α. KEGG analysis highlighted the PI3K/AKT signaling pathway as a key mediator of BBR's anti-RA effects. Metabolomics profiling via LC-MS revealed 22 potential biomarkers. Arginine and proline metabolism, cutin, suberine and wax biosynthesis, glycine, serine and threonine metabolism and taurine and hypotaurine metabolism are the most related pathways of BBR anti-RA. Molecular docking studies corroborated high affinities between BBR and key targets. Furthermore, 16S analysis demonstrated BBR's capacity to modulate gut bacteria composition, including an increase in the abundance of Lachnoclostridium, Akkermansia, Blautia, Romboutsia, and Faecalibacterium genera, alongside a decrease in Prevotella_9 abundance in genus level. Integrated analysis underscored a strong correlation between serum microbiota and fecal metabolites. Our findings elucidate the multifaceted mechanisms underlying BBR's therapeutic efficacy in RA, involving inhibition of the PI3K/AKT pathway, modulation of intestinal flora, and regulation of host metabolites. These insights provide novel perspectives on BBR's role in RA management.

Regulation of Tumor Growth and Invasion in Oral Squamous Cell Carcinoma by the Tumor Microenvironment Through the Enhancement of IGFBP-3 Expression.

Accumulated evidence indicates that interactions among various stromal cells within the tumor microenvironment (TME) significantly influence cancer progression. Oral cancers not diagnosed at early stages are associated with low five-year survival rates, highlighting the need for substantial improvements in patient outcomes. Understanding the interactions between cancer cells and the tumor microenvironment is crucial for identifying methods and developing treatment strategies that more effectively inhibit tumor progression and metastasis. This study investigated the roles of cancer-associated fibroblasts (CAFs) and THP-1 monocytes in tumor growth and invasion, observing changes in cancer biological characteristics through interactions with stromal cells. HSC2 oral squamous cell carcinoma (OSCC) cells were co-cultured with normal fibroblasts (NF), cancer-associated fibroblasts (CAF), or THP-1 cells. Changes in cancer cell invasion were investigated using Matrigel-coated transwell assays, collagen-based dermal equivalent assays, and in vivo mouse studies. HSC2 cells co-cultured with CAFs or THP-1 exhibited increased invasion compared to those co-cultured with normal fibroblasts (NF) in the dermal equivalent. In a mouse gingival xenograft model, cancer cells co-implanted with CAFs or THP-1 showed significantly increased tumor growth compared to those co-implanted with NF. Micro-CT (μCT) analysis revealed significant alveolar bone resorption around the mandible in tumors grown with CAFs or THP-1 cells. Conditioned media (CM) from CAFs or THP-1 significantly increased HSC2 invasion and migration, an effect that was inhibited by the protein secretion inhibitor Brefeldin (BFA). Furthermore, QuantSeq 3' mRNA sequencing indicated increased expression of IGFBP3, closely associated with cancer invasion, in HSC2 cells co-cultured with CAFs or THP-1. These findings suggest that cancer cells enhance their invasive characteristics through close interactions with the surrounding stromal cells.