Detecting and Quantifying Xylem Embolism by Synchrotron-Based X-Ray Micro-CT.

The vulnerability to xylem embolism is a key trait underlying species-specific drought tolerance of plants, and hence is critical for screening climate-resilient crops and understanding vegetation responses to drought and heat waves. Yet, accurate determination of embolism in plant's xylem is challenging, because most traditional hydraulic techniques are destructive and prone to artefacts. Hence, direct and in vivo synchrotron-based X-ray micro-CT observation of xylem conduits has emerged as a key reference technique for accurate quantification of vulnerability to xylem embolism. Micro-CT is nowadays a fundamental tool for studies of plant hydraulic architecture, and this chapter describes the fundamentals of acquisition and processing of micro-CT images of plant xylem.

Yunvjian decoction attenuates lipopolysaccharide-induced periodontitis by suppressing NFκB/NLRP3/IL-1ß pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Yunvjian decoction (YNJ) is a traditional Chinese herbal prescription that has been used in the clinical treatment of periodontitis. However, the underlying molecular mechanism of YNJ in the periodontitis treatment is not well understood. AIM OF THE STUDY: The purpose of this study was to evaluate the therapeutic effects of YNJ against periodontitis and its underlying molecular mechanisms. MATERIALS AND METHODS: Orthodontic ligation and lipopolysaccharide (LPS)-induced periodontitis rat model was established. YNJ groups were gavaged with YNJ decoction (5 g/kg/d or 10 g/kg/d) for four months. The rats in positive control group were gavaged with metronidazole (MDZ, 100 mg/kg/d) for four months. The maxilla was scanned by micro-computed tomography. The chemical compositions of YNJ were identified using ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry. The molecular mechanism of YNJ were predicted using network pharmacological analysis and validated using immune-staining and Western blot. RESULTS: YNJ treatment decreased the distance between cementoenamel junction and alveolar bone crest on the sagittal slide of the periodontitis rats. Western blot showed YNJ downregulated the protein levels of the bone resorption marker (receptor activator of nuclear factor-κB ligand), while upregulated the levels of the bone formation markers (bone morphogenetic protein 2, runt-related transcription factor 2, alkaline phosphatase, and osteoprotegerin) in alveolar bone of the periodontitis rats. Hematoxylin and eosin, immunohistochemical staining, and Western blot analysis indicated that YNJ attenuated the inflammation and decreased the levels of interleukin-6 and tumor necrosis factor-α in the alveolar bone. In addition, a total of 61 compounds were identified from YNJ. Network pharmacology indicated that the nucleotide binding oligomerization domain-like receptor signaling pathway was the main pathway for YNJ in the treatment of periodontitis. The experiments confirmed that YNJ administration inhibited LPS induced-pyroptosis in alveolar bone through suppressing the phosphorylation of nuclear factor κB, reduced expression of NOD-like receptor family pyrin domain containing 3, and Caspase-1, subsequently suppressing the interleukin-1ß secretion. CONCLUSION: YNJ is an effective therapeutic strategy for periodontitis and acts by inhibiting pyroptosis and NFκB/NLRP3/IL-1ß pathway in alveolar bone.

Small animal DXA instrument comparison and validation.

Several new peripheral dual-energy X-ray absorptiometry (DXA) devices designed for assessment of bone and body composition in rodents have been developed. We compared the performance (accuracy and precision) of two of these devices, the InAlyzer and the iNSiGHT, to those of an established device, the PIXImus. We measured total body bone mineral content (BMC), bone mineral density (BMD), and body composition (lean and fat mass) on the three DXA devices in 18 male C57Bl/6 J mice (6 each of ages 8, 14, and 24 weeks, weighing 22 to 33 g). DXA body composition measures were compared to whole-body nuclear magnetic resonance (NMR) outcomes. BMC of the femur was also compared to ex vivo micro-computed tomography (microCT). Total body BMD from the InAlyzer and iNSiGHT devices was strongly correlated to that from PIXImus (R2 = 0.83 and 0.82, respectively), but was ~25 % higher than PIXImus. Total body BMC measures by InAlyzer were strongly associated with those from PIXImus (R2 = 0.86), whereas those from iNSiGHT were only weakly correlated (R2 = 0.29). Femur BMC from InAlyzer was strongly correlated with microCT outcomes, whereas iNSiGHT was only weakly correlated. InAlyzer and iNSiGHT fat mass measures were very strongly correlated with PIXImus and NMR outcomes (R2 = 0.91 to 0.97), with slightly weaker associations for lean mass (R2 = 0.81 to 0.76). Short-term precision of InAlyzer and iNSiGHT measurements were excellent, and akin to those from the PIXImus for both body composition and bone measures, ranging between 0.39 and 3.2 %. With faster scan times, closed X-ray source and excellent precision, the new devices are both satisfactory replacements for the now discontinued PIXImus system. However, given the accuracy of the bone and body composition measures, the InAlyzer may be preferable for studies where musculoskeletal changes are the main interest.

The sacroiliac joint: An original and highly sensitive tool to highlight altered bone phenotype in murine models of skeletal disorders.

Bone disorders may affect the skeleton in different ways, some bones being very impaired and others less severely. In translational studies using murine models of human skeletal diseases, the bone phenotype is mainly evaluated at the distal femur or proximal tibia. The sacroiliac joint (SIJ), which connects the spine to the pelvis, is involved in the balanced transfer of mechanical energy from the lumbar spine to the lower extremities. Because of its role in biomechanical stress, the SIJ is a region of particular interest in various bone diseases. Here we aimed to characterize the SIJ in several murine models to develop a highly reliable tool for studying skeletal disorders. We performed a 12-month in vivo micro-computed tomography (micro-CT) follow-up to characterize the SIJ in wild-type (WT) C57BL/J6 mice and compared the bone microarchitecture of the SIJ and the distal femur at 3 months by micro-CT and histology. To test the sensitivity of our methodology, the SIJ and distal femur were evaluated at 3 and 6 months, in 2 murine models of skeletal disorder, X-linked hypophosphatemia (Hyp mice) and HLA-B27 transgenic mice and compared to WT mice. A multimodal analysis was performed, using a combination of microCT and histological analysis. With the Hyp model, the SIJ displayed more bone microarchitecture alterations than the distal femur. Hyp mice showed a significant reduction in trabecular bone at both the distal femur and sacral slope as compared with WT mice, with a significant positive correlation between trabecular bone parameters of the distal femur and sacral side of the SIJ. Furthermore, trabecular bone parameters (Bone Volume/Total Volume (BV/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular number (Tb.N), trabecular pattern factor (Tb.Pf)) were significantly increased compared to femoral parameters at the SIJ. The sacral articular cortical bone, which is indicative of osteoarticular lesions, was altered in Hyp mice. Interestingly, in accordance to previous studies, HLA-B27 transgenic mice did not show any osteoarticular lesions as compared with WT mice. Cortical bone parameters (thickness, porosity), as well as scoring performed with double blinding, did not show difference between the 2 genotypes. The characterization and evaluation of the SIJ surface appears very sensitive to emphasize alterations of bone and joint. The SIJ may represent a valuable tool to investigate both bone and local osteoarticular alterations in murine models of skeletal disorders and might be a relevant site for assessing the response to treatment of chronic bone diseases.

Phage PEf771 for the Treatment of Periapical Periodontitis Induced by Enterococcus faecalis YN771.

Enterococcus faecalis was the main causative bacteria of refractory periapical periodontitis (PP), there is a pressing need to explore effective methods for eradicating E. faecalis in patients with refractory PP. This study aimed to assess the anti-infective effectiveness of phage PEf771 in treating periapical inflammation in rats. We developed a rat model of PP through E. faecalis YN771 induction. Micro-computed tomography and hematoxylin-eosin staining were utilized to evaluate bone destruction and inflammation in experimental teeth for seven consecutive weeks. Subsequently, rats with PP caused by E. faecalis YN771 were treated with phage PEf771, calcium hydroxide preparation, and 2% chlorhexidine gel. The healing progress of bone defects and inflammation in the apical region was monitored over three consecutive weeks using imaging and histopathology assessments. The PP rat model was successfully developed, and bone destruction and inflammatory cell infiltration in the apical region of the experimental tooth peaked at 4 weeks. The area of bone destruction in rats treated with phage PEf771, calcium hydroxide preparation, and 2% chlorhexidine gel was significantly smaller than that in the untreated group. Phage PEf771, calcium hydroxide preparation, and 2% chlorhexi-dine gel all have the effect of promoting the healing of apical lesions. Therapeutic effects of phage PEf771 on periapical inflammation infected by E. faecalis YN771 enhanced with time. Phage PEf771 promoted the healing of apical lesions, presenting a promising new approach for the treatment of refractory PP using bacteriophages.

Development of a membrane and a bilayer of chitosan, gelatin, and polyhydroxybutyrate to be used as wound dressing for the regeneration of rat excisional wounds.

The skin is the largest organ in the human body that acts as a protective barrier from the outside environment. Certain dermatological pathologies or significant skin lesions can result in serious complications. Several studies have focused on the development of tissue-engineered skin substitutes. In this study, a new bilayer scaffold composed of a chitosan-gelatin membrane and a chitosan-polyhydroxybutyrate (PHB) porous matrix was synthesized and populated with human adipose-derived mesenchymal stem cells (hASCs) to be potentially used for wound dressing applications. By combining this membrane and porous matrix with the stem cells, we aimed to provide immunomodulation and differentiation capabilities for the wound environment, as well as mechanical strength and biocompatibility for the underlying tissue. The membrane was prepared from the mixture of chitosan and gelatin in a 2:1 ratio and the porous matrix was prepared from the mixture of chitosan and PHB, in equal proportions to form a final solution at 2.5% (m/v). Fourier transform infrared spectroscopy analysis showed the formation of blends, and micro-computed tomography, scanning electron microscopy and atomic force microscopy images demonstrated membrane roughness and matrix porosity. The MTT assay showed that the scaffolds were biocompatible with hASC. The membrane and the bilayer were used as dressing and support for cell migration in the dorsal excisional wound model in Wistar rats. Histological and gene transcriptional analyses showed that the animals that received the scaffolds regenerated the hair follicles in the deep dermis in the central region of the wound. Our results demonstrate the potential of these new biomaterials as dressings in wound healing studies, favoring tissue regeneration.

Flavonoid extract from propolis alleviates periodontitis by boosting periodontium regeneration and inflammation resolution via regulating TLR4/MyD88/NF-κB and RANK/NF-κB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: In traditional Chinese medicine, propolis has been used for treating oral diseases for centuries, widely. Flavonoid extract is the main active ingredient in propolis, which has attracted extensive attention in recent years. AIM OF THE STUDY: The objective and novelty of the current study aims to identify the mechanism of total flavonoid extract of propolis (TFP) for the treatment of periodontitis, and evaluate the therapeutic effect of TFP-loaded liquid crystal hydrogel (TFP-LLC) in rats with periodontitis. METHODS: In this study, we used lipopolysaccharide-stimulated periodontal ligament stem cells (PDLSCs) to construct in vitro inflammation model, and investigated the anti-inflammatory effect of TFP by expression levels of inflammatory factors. Osteogenic differentiation was assessed using alkaline phosphatase activity and alizarin red staining. Meanwhile, the expression of toll like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), nuclear factor-kappa B (NF-κB), receptor activator of NF-κB (RANK) etc, were quantitated to investigate the therapeutic mechanism of TFP. Finally, we constructed TFP-LLC using a self-emulsification method and administered it to rats with periodontitis via periodontal pocket injection to evaluate the therapeutic effects. The therapeutic index, microcomputed tomography (Micro-CT), H&E staining, TRAP staining, and Masson staining were used for this evaluation. RESULTS: TFP reduced the expression of TLR4, MyD88, NF-κB and inflammatory factor in lipopolysaccharide-stimulated PDLSCs. Meanwhile, TFP simultaneously regulating alkaline phosphatase, RANK, runt-associated transcription factor-2 and matrix metalloproteinase production to accelerate osteogenic differentiation and collagen secretion. In addition, TFP-LLC can stably anchor to the periodontal lesion site and sustainably release TFP. After four weeks of treatment with TFP-LLC, we observed a decrease in the levels of NF-κB and interleukin-1ß (IL-1ß) in the periodontal tissues of rats, as well as a significant reduction in inflammation in HE staining. Similarly, Micro CT results showed that TFP-LLC could significantly inhibit alveolar bone resorption, increase bone mineral density (BMD) and reduce trabecular bone space (Tb.Sp) in rats with periodontitis. CONCLUSION: Collectively, we have firstly verified the therapeutic effects and mechanisms of TFP in PDLSCs for periodontitis treatment. Our results indicate that TFP perform anti-inflammatory and tissue repair activities through TLR4/MyD88/NF-κB and RANK/NF-κB pathways in PDLSCs. Meanwhile, for the first time, we employed LLC delivery system to load TFP for periodontitis treatment. The results showed that TFP-LLC could be effectively retained in the periodontal pocket and exerted a crucial role in inflammation resolution and periodontal tissue regeneration.

Crossing fibers may underlie the dynamic pulling forces of muscles that attach to cartilage at the tip of the nose.

The present study used microdissection, histology, and microcomputed tomography (micro-CT) with the aims of determining the prevalence and patterns of the depressor septi nasi (DSN) and orbicularis oris (OOr) muscles attached to the footplate of the medial crus (fMC) of the major alar cartilage, focusing on their crossing fibers. The DSN and OOr attached to the fMC of the major alar cartilage were investigated in 76 samples from 38 embalmed Korean adult cadavers (20 males, 18 females; mean age 70 years). The DSN, OOr, or both were attached to the fMC. When the DSN ran unilaterally or was absent, some OOr fibers ascended to attach to the fMC instead of the DSN in 20.6% of the samples. Crossing fibers of the DSN or OOr attached to the fMC were found in 82.4% of the samples. Bilateral and unilateral crossing fibers were found in 32.4% and 50.0%, respectively, and no crossing fibers were found in 17.6%. The DSN and OOr that attached to the fMC could be categorized into six types according to presence of the DSN and the crossing patterns of the DSN and OOr. Anatomical findings of the DSN and OOr that attached to the fMC were confirmed in histology and micro-CT images. These findings offer insights on anatomical mechanisms that may underlie the dynamic pulling forces generated by muscles that attach to the fMCs and on evolutionary variation observed in human facial expressions. They can also provide useful information for guiding rhinoplasty of the nasal tip.

The Establishment of a Murine Maxillary Orthodontic Model.

Due to the lack of reproducible protocols for establishing a murine maxillary orthodontic model, we present a reliable and reproducible protocol to provide researchers with a feasible tool to analyze mechanical loading-associated bone remodeling. This study presents a detailed flowchart in addition to different types of schematic diagrams, operation photos, and videos. We performed this protocol on 11 adult wide-type C57/B6J mice and harvested samples on postoperative days 3, 8, and 14. The micro-CT and histopathological data have proven the success of tooth movements coupled with bone remodeling using this protocol. Furthermore, according to the micro-CT results on days 3, 8, and 14, we have divided bone modeling into three stages: preparation stage, bone resorption stage, and bone formation stage. These stages are expected to help researchers concerned with different stages to set sample collection time reasonably. This protocol can equip researchers with a tool to carry out regenerative analysis of bone remodeling.

Erythropoietin suppresses osteoblast apoptosis and ameliorates steroid-induced necrosis of the femoral head in rats by inhibition of STAT1-caspase 3 signaling pathway.

BACKGROUND: Steroid-induced avascular necrosis of the femoral head (SANFH) is characterized by osteoblast apoptosis, leading to a loss of bone structure and impaired hip joint function. It has been demonstrated that erythropoietin (EPO) performs a number of biological roles. OBJECTIVE: We examined the effects of EPO on SANFH and its regulation of the STAT1-caspase 3 signaling pathway. METHOD: In vitro, osteoblasts were treated with dexamethasone (Dex) or EPO. We identified the cytotoxicity of EPO by CCK-8, the protein expression of P-STAT1, cleaved-caspase9, cleaved-caspase3, Bcl-2, BAX, and cytochrome c by Western blotting, and evaluated the apoptosis of osteoblasts by flow cytometry. In vivo, we analyzed the protective effect of EPO against SANFH by hematoxylin and eosin (H&E), Immunohistochemical staining, and Micro-computed tomography (CT). RESULTS: In vitro, EPO had no apparent toxic effect on osteoblasts. In Dex-stimulated cells, EPO therapy lowered the protein expression of BAX, cytochrome c, p-STAT1, cleaved-caspase9, and cleaved-caspase3 while increasing the expression of Bcl-2. EPO can alleviate the apoptosis induced by Dex. In vivo, EPO can lower the percentage of empty bone lacunae in SANFH rats. CONCLUSION: The present study shows that EPO conferred beneficial effects in rats with SANFH by inhibiting STAT1-caspase 3 signaling, suggesting that EPO may be developed as a treatment for SANFH.

Efficiency and complications in root canal retreatment using nickel titanium rotary file with continuous rotation, reciprocating, or adaptive motion in curved root canals: a laboratory investigation.

BACKGROUND: It is currently unknown whether rotary file motion affects the best outcome of root canal retreatment. This experimental study compared the efficacy, efficiency, and complications of single-use NiTi rotary files using continuous rotation, reciprocating, and adaptive motions in root canal filling removal in curved root canals. Reciproc blue R25 was used with reciprocating motion (RB), VDW.ROTATE retreatment files with continuous rotation (VR), and ProTaper NEXT X2 with continuous rotation (PTNc) or adaptive motion (PTNa). METHODS: Forty mesial root canals of extracted mandibular first and second molars with an angle of curvature between 20°-40° and a radius of curvature between 5 and 10 mm were collected. The specimens were instrumented and obturated with gutta-percha and AH Plus sealer using the continuous wave of condensation technique. The specimens were randomly divided into 4 retreatment groups (n = 10), RB, VR, PTNc, and PTNa. The percentage of root canal filling removal in each group was analyzed using Micro-Computed Tomography (µCT). The motor running time, total time, root canal complication, and instrument complication were recorded and statistically analyzed (p-value < 0.05). RESULTS: The pre-operative root canal curvature and root canal filling volume were comparable among groups. The percentage of root canal filling removal from the whole canal in the PTNc, RB, PTNa, and VR group was 98%, 96%, 95%, and 93%, respectively. A significant difference was observed between the PTNc and VR groups for the whole canal and the apical-third part. The motor running time and total time were significantly different between the groups. Instrument fracture was observed at 40% in the VR and 20% in the PTNa group, but none in the RB and PTNc groups. CONCLUSIONS: The ProTaper NEXT X2 with continuous rotation and RB files can be used with high efficacy and efficiency in curved root canal retreatment. Continuous rotation is more efficacious and efficient than adaptive motion when using the NiTi rotary file. Single file retreatment can be used in small canals with high efficacy, cost-effectiveness, and less time consumption.

New ex vivo method to objectively assess insulin spatial subcutaneous dispersion through time during pump basal-rate based administration.

Glycemic variability remains frequent in patients with type 1 diabetes treated with insulin pumps. Heterogeneous spreads of insulin infused by pump in the subcutaneous (SC) tissue are suspected but were barely studied. We propose a new real-time ex-vivo method built by combining high-precision imaging with simultaneous pressure measurements, to obtain a real-time follow-up of insulin subcutaneous propagation. Human skin explants from post-bariatric surgery are imaged in a micro-computed tomography scanner, with optimised parameters to reach one 3D image every 5 min during 3 h of 1UI/h infusion. Pressure inside the tubing is recorded. A new index of dispersion (IoD) is introduced and computed upon the segmented 3D insulin depot per time-step. Infusions were hypodermal in 58.3% among 24 assays, others being intradermal or extradermal. Several minor bubbles and one occlusion were observed. IoD increases with time for all injections. Inter-assay variability is the smallest for hypodermal infusions. Pressure elevations were observed, synchronised with air bubbles arrivals in the tissue. Results encourage the use of this method to compare infusion parameters such as pump model, basal rate, catheter characteristics, infusion site characteristics or patient phenotype.

Efficacy of bone defect therapy involving various surface treatments of titanium alloy implants: an in vivo and in vitro study.

Multiple surface treatment methods for titanium alloy prostheses, widely used in orthopedics, are available; however, these can affect bone integration and regeneration efficiency. In this study, through cell and animal experiments, we devised seven bone implant categories of Ti6Al4V based on surface preparation and post-processing technology (polishing, grit-blasting, fine titanium spraying, coarse titanium spraying, electron beam melting [EBM] printing, selective laser melting [SLM] printing, and post-processed SLM printing) and imaged each microscopic surface structure with a scanning electron microscope (SEM). Mechanical testing revealed excessive post-processing damaged the mechanical properties of the implants. In vitro, human bone marrow mesenchymal stem cells (hBMSCs) were cultured with implants, and the morphology of the cells adhering to the implant surface was observed using SEM and confocal laser scanning microscopy. Cell Counting Kit-8 (CCK-8) semi-quantitatively determined cell activity, indirectly reflecting the proliferation of hBMSCs. Alizarin red and alkaline phosphatase experiments assessed osteogenic differentiation. In vivo, experiments utilized the New Zealand rabbit femoral condyle bone defect model to assess bone regeneration and integration using micro-computed tomography, Van Giesen staining, and Masson staining. We found that 3D-printed implants with regular pore structures were more conducive to hBMSC osteogenic differentiation, while the presence of metal powder on NPT-SLM-printed implants hindered such differentiation. The post-treatment SLM scaffold surface may have some residual semi-melted powder; however, these powder residues have no significant effect on cell activity and differentiation. Surface treatment (grit-blasting and titanium spraying) of planar structures can enhance hBMSC adhesion but does not necessarily promote their differentiation. The framework structure of 3D printing may affect the osteogenic differentiation of hBMSCs, and for SLM-printed implants, excessive pursuit of a "powderless" state will damage the mechanical properties of the implant.

Simulation study of a novel small animal FLASH irradiator (SAFI) with integrated inverse-geometry CT based on circularly distributed kV X-ray sources.

Ultra-high dose rate (UHDR) radiotherapy (RT) or FLASH-RT can potentially reduce normal tissue toxicity. A small animal irradiator that can deliver FLASH-RT treatments similar to clinical RT treatments is needed for pre-clinical studies of FLASH-RT. We designed and simulated a novel small animal FLASH irradiator (SAFI) based on distributed x-ray source technology. The SAFI system comprises a distributed x-ray source with 51 focal spots equally distributed on a 20 cm diameter ring, which are used for both FLASH-RT and onboard micro-CT imaging. Monte Carlo simulation was performed to estimate the dosimetric characteristics of the SAFI treatment beams. The maximum dose rate, which is limited by the power density of the tungsten target, was estimated based on finite-element analysis (FEA). The maximum DC electron beam current density is 2.6 mA/mm2, limited by the tungsten target's linear focal spot power density. At 160 kVp, 51 focal spots, each with a dimension of [Formula: see text] mm2 and 10° anode angle, can produce up to 120 Gy/s maximum DC irradiation at the center of a cylindrical water phantom. We further demonstrate forward and inverse FLASH-RT planning, as well as inverse-geometry micro-CT with circular source array imaging via numerical simulations.

Mice Lacking PLAP-1/Asporin Show Alteration of Periodontal Ligament Structures and Acceleration of Bone Loss in Periodontitis.

Periodontal ligament-associated protein 1 (PLAP-1), also known as Asporin, is an extracellular matrix protein expressed in the periodontal ligament and plays a crucial role in periodontal tissue homeostasis. Our previous research demonstrated that PLAP-1 may inhibit TLR2/4-mediated inflammatory responses, thereby exerting a protective function against periodontitis. However, the precise roles of PLAP-1 in the periodontal ligament (PDL) and its relationship to periodontitis have not been fully explored. In this study, we employed PLAP-1 knockout mice to investigate its roles and contributions to PDL tissue and function in a ligature-induced periodontitis model. Mandibular bone samples were collected from 10-week-old male C57BL/6 (WT) and PLAP-1 knockout (KO) mice. These samples were analyzed through micro-computed tomography (µCT) scanning, hematoxylin and eosin (HE) staining, picrosirius red staining, and fluorescence immunostaining using antibodies targeting extracellular matrix proteins. Additionally, the structure of the PDL collagen fibrils was examined using transmission electron microscopy (TEM). We also conducted tooth extraction and ligature-induced periodontitis models using both wild-type and PLAP-1 KO mice. PLAP-1 KO mice did not exhibit any changes in alveolar bone resorption up to the age of 10 weeks, but they did display an enlarged PDL space, as confirmed by µCT and histological analyses. Fluorescence immunostaining revealed increased expression of extracellular matrix proteins, including Col3, BGN, and DCN, in the PDL tissues of PLAP-1 KO mice. TEM analysis demonstrated an increase in collagen diameter within the PDL of PLAP-1 KO mice. In line with these findings, the maximum stress required for tooth extraction was significantly lower in PLAP-1 KO mice in the tooth extraction model compared to WT mice (13.89 N ± 1.34 and 16.51 N ± 1.31, respectively). In the ligature-induced periodontitis model, PLAP-1 knockout resulted in highly severe alveolar bone resorption, with a higher number of collagen fiber bundle tears and significantly more osteoclasts in the periodontium. Our results demonstrate that mice lacking PLAP-1/Asporin show alteration of periodontal ligament structures and acceleration of bone loss in periodontitis. This underscores the significant role of PLAP-1 in maintaining collagen fibrils in the PDL and suggests the potential of PLAP-1 as a therapeutic target for periodontal diseases.

Insertion methods and gap/void formation in atraumatic restorative technique: A micro-CT analysis.

Wetting the dentin is critical to atraumatic restorative treatment. The conventional insertion can be challenging when using high-viscosity glass-ionomer cement. This study evaluated the formation of gaps and voids after three insertion methods using micro-CT. Teeth underwent removal of occlusal and proximal caries through the atraumatic restorative treatment technique. Then, they were fixed in an artificial dental arch to simulate the clinical condition and were restored using three insertion methods: conventional, Centrix injection, and double-filling. Previous dentin conditioning procedures, steel matrix and wooden wedge application, and post-insertion procedures (digital compression and surface protection) were the same. The material was inserted using a manual instrument in the conventional technique and was inserted with a syringe in the Centrix injection group. In the double-filling technique, the tooth received a first layer of a flowable ionomer (through modifying the powder/liquid ratio), and a second layer (with standard ratio) was applied before the final set of the first one. A micro-CT unit scanned each tooth before and after restoration. Each cavity was defined as the volume of interest, and the volumes of gaps and voids were calculated. Data were analyzed using one-way ANOVA and Tukey posthoc test (p < .05). Double-filling had improved filling volume with lower values for gap volume, followed by Centrix injection. The conventional technique had the highest gap volume. No statistically significant difference was observed for void volume. Double-filling demonstrated fewer gaps, followed by Centrix injection, which is critical to obtain better adhesive, remineralizing, and antibacterial activities.

Parathyroid hormone stimulates bone regeneration in an atrophic non-union model in aged mice.

BACKGROUND: Non-union formation still represents a major burden in trauma and orthopedic surgery. Moreover, aged patients are at an increased risk for bone healing failure. Parathyroid hormone (PTH) has been shown to accelerate fracture healing in young adult animals. However, there is no information whether PTH also stimulates bone regeneration in atrophic non-unions in the aged. Therefore, the aim of the present study was to analyze the effect of PTH on bone regeneration in an atrophic non-union model in aged CD-1 mice. METHODS: After creation of a 1.8 mm segmental defect, mice femora were stabilized by pin-clip fixation. The animals were treated daily with either 200 mg/kg body weight PTH 1-34 (n = 17) or saline (control; n = 17) subcutaneously. Bone regeneration was analyzed by means of X-ray, biomechanics, micro-computed tomography (µCT) imaging as well as histological, immunohistochemical and Western blot analyses. RESULTS: In PTH-treated animals bone formation was markedly improved when compared to controls. This was associated with an increased bending stiffness as well as a higher number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts and CD31-positive microvessels within the callus tissue. Furthermore, PTH-treated aged animals showed a decreased inflammatory response, characterized by a lower number of MPO-positive granulocytes and CD68-positive macrophages within the bone defects when compared to controls. Additional Western blot analyses demonstrated a significantly higher expression of cyclooxygenase (COX)-2 and phosphoinositide 3-kinase (PI3K) in PTH-treated mice. CONCLUSION: Taken together, these findings indicate that PTH is an effective pharmacological compound for the treatment of non-union formation in aged animals.

Effect of intra-articular injection of a hyaluronic acid-alendronate conjugate on post-traumatic osteoarthritis induced by destabilization of the medial meniscus in rats.

Osteoarthritis (OA) is a chronic degenerative joint disease characterized by pain and cartilage damage. Intra-articular (i.a) viscosupplementation with hyaluronic acid (HA) is frequently used for the management of OA. Preclinical studies have reported that bisphosphonates (BPs) may have a therapeutic potential to slow down or reverse the progression of OA. Among these, alendronate (ALN) has demonstrated chondroprotective effects in both in vitro and vivo experiments. This study evaluated the effects of a novel alendronate-hyaluronic acid (ALN-HA) conjugate on an OA in vivo model induced by medial meniscus destabilization (DMM). DMM surgery was performed on the knees of Sprague Dawley rats that received, after four weeks, one intra-articular (i.a.) injection of: (1) ALN-HA; (2) HA; (3) sodium chloride (NaCl). Sham-operated rats were used as control. Allodynia was assessed by Von Frey test. Joint degeneration was evaluated eight weeks after treatment by micro-computed tomography (micro-CT), histology, and immunohistochemistry. Collagen cross-linked C-telopeptides (CTX-I and CTX-II) serum levels were determined by ELISA. Paw withdrawal threshold increased in ALN-HA group when compared to rats treated with NaCl or HA. Micro-CT did not show differences between ALN-HA, HA and NaCl groups. ALN-HA injection produced significant improvements in articular cartilage degeneration showing an OARSI score lower than those of HA and NaCl, and reduced matrix metalloproteinase (MMP)-13, MMP-3, interleukin-6, vascular endothelial growth factor and Caspase-3 expression. CTX-I was reduced after ALN-HA treatment when compared to NaCl. Our results indicate that i.a. use of ALN after conjugation with HA limits OA development and progression in the rat DMM model, and may lead to the development of novel therapeutic strategies in OA management.

Subcortical life, evolution of flattened body, and constrained mating posture in the earwig Platylabia major (Insecta: Dermaptera: "Anisolabididae").

Many animals take advantage of the shaded, humid, and protected environments in subcortical spaces, i.e., thin spaces under the loosened bark of dead trees. Permanent inhabitants of subcortical spaces often show specialized morphologies, such as a miniaturized or dorsoventrally flattened body. However, the evolutionary consequences of these specialized morphologies on behavioral, ecological, and life-history traits have been little studied. We studied the mating biology and anatomy of Platylabia major (usually placed in the family Anisolabididae), which is an obligate inhabitant of subcortical spaces with a paper-like flattened body, and compared them with those of two thicker, spongiphorid earwigs, Nesogaster amoenus and Paralabellula curvicauda. Mating trials in various settings showed that Pl. major requires thin spaces sandwiched by two planes to accomplish genital coupling and insemination. In contrast, the thicker species, although also frequently found in subcortical spaces, could mate on a single horizontal plane due to the ability of the male to twist its abdomen through approximately 180°. Examination by micro-computed tomography and a reagent-based clearing technique revealed no substantive differences in the configuration of mid-abdominal musculature between the species. The dorsal and lateral muscles of Pl. major, which are almost parallel to the antero-posterior body axis for accommodation within the thin abdomen, seemed incapable of producing the power to twist the abdomen. The abdominal musculature conforms to a simple pattern in both male and female earwigs, which is repeated in each of the pregenital segments. We conclude that small differences in the range of motion of each abdominal segment can result in large differences in possible mating postures and positions. Surgical experiments also demonstrated that both right and left penises of Pl. major are competent and used for insemination with no lateral bias, as in most other earwigs with twin penises studied to date.

A longitudinal rat model for assessing postoperative recovery and bone healing following tibial osteotomy and plate fixation.

BACKGROUND: Rodent models are commonly employed to validate preclinical disease models through the evaluation of postoperative behavior and allodynia. Our study investigates the dynamic interplay between pain and functional recovery in the context of traumatic osteotomy and surgical repair. Specifically, we established a rat model of tibial osteotomy, followed by internal fixation using a 5-hole Y-plate with 4 screws, to explore the hypothesis that histological bone healing is closely associated with functional recovery. OBJECTIVE: Our primary objective was to assess the correlation between bone healing and functional outcomes in a rat model of tibial osteotomy and plate fixation. METHODS: Seventeen male Sprague-Dawley rats underwent a metaphyseal transverse osteotomy of the proximal tibia, simulating a fracture-like injury. The resultant bone defect was meticulously repaired by realigning and stabilizing the bone surfaces with the Y-plate. To comprehensively assess recovery and healing, we performed quantitative and qualitative evaluations at 2, 4, 6, and 8 weeks post-surgery. Evaluation methods included micro-CT imaging, X-ray analysis, and histological examination to monitor bone defect healing. Concurrently, we employed video recording and gait analysis to evaluate functional recovery, encompassing parameters such as temporal symmetry, hindlimb duty factor imbalance, phase dispersion, and toe spread. RESULTS: Our findings revealed complete healing of the bone defect at 8 weeks, as confirmed by micro-CT and histological assessments. Specifically, micro-CT data showed a decline in fracture volume over time, indicating progressive healing. Histological examination demonstrated the formation of new trabecular bone and the resolution of inflammation. Importantly, specific gait analysis parameters exhibited longitudinal changes consistent with bone healing. Hindlimb duty factor imbalance, hindlimb temporal symmetry, and phase dispersion correlated strongly with the healing process, emphasizing the direct link between bone healing and functional outcomes. CONCLUSIONS: The establishment of this tibia osteotomy model underscores the association between bone healing and functional outcomes, emphasizing the feasibility of monitoring postoperative recovery using endpoint measurements. Our overarching objective is to employ this model for assessing the local efficacy of drug delivery devices in ameliorating post-surgical pain and enhancing functional recovery.