micro-CT Analysis Research Articles

Fibroblast growth factor 7 (FGF7) causes cartilage destruction, subchondral bone remodeling, and the premature growth plate closure in mice

ObjectiveFibroblast growth factor (FGF) signaling plays a significant role in osteoarthritis (OA) pathogenesis, though the OA-related functions of only a few FGFs have been fully elucidated. This study investigates the specific roles of FGF7 in OA development. MethodsFGF7 expression was analyzed in human (n=6) and mouse (n=10) cartilage. Experimental OA was induced by destabilization of the medial meniscus (DMM). The roles of FGF7 were explored using intra-articular (IA) injection of recombinant FGF7 (rFGF7) and whole-body Fgf7 knockout mice (Fgf7-/-). Subchondral bone remodeling and growth plate morphology were assessed microCT and histological analysis. ResultsFGF7 was upregulated in OA cartilage. IA injection of rFGF7 led to OA cartilage destruction (OARSI grade; 0.61 [95% CI 0.00–5.33]), while Fgf7-/- mice showed reduced DMM-induced cartilage erosion (OARSI grade; 1.89 [95% CI 1.08–3.00]) compared to WT mice (4.92 [95% CI 3.83–5.33]). These effects were associated with changes in matrix-degrading enzyme expression in chondrocytes. Mice receiving IA injection of rFGF7 (20 μg) exhibited increased subchondral bone thickness (68.01 μm [95% CI 61.55–74.46]) and decreased osteoclastogenesis (TRAP positivity; 1.94% [95% CI 1.41–2.47]) compared to controls (38.33 μm [95% CI 33.71–42.96]) and (4.23% [95% CI 3.28–5.19]), respectively. Additionally, rFGF7 treatment caused premature closure of growth plates, whereas Fgf7-/- mice exhibited significantly increased growth plate thickness. ConclusionsFGF7 exerts multiple functions in various joint tissues, including promoting cartilage destruction, inducing subchondral bone remodeling (SBP thickening), and triggering premature growth plate closure.

Bone regeneration in rabbit cranial defects: 3D printed polylactic acid scaffolds gradually enriched with marine bioderived calcium phosphate

ObjectiveThis study aimed to evaluate the in vivo biocompatibility, mechanical performance and osteoconductive potential of 3D-printed polylactic acid (PLA) scaffolds enriched with marine bioderived calcium phosphate (bioCaP) for bone tissue engineering. Materials and MethodsPLA-bioCaP composite scaffolds were specifically designed for the rabbit cranial defect model by 3D printing, with a uniform distribution of open square-shaped pores and contributions in bioCaP. Physicochemical and mechanical characterization and the evaluation of biological response are presented. ResultsThe scaffolds demonstrated mechanical properties comparable to human bones, integration with the host bone, and osteoconductive behavior promoting cell ingrowth from the defect edge. Strong mineralized tissue ingrowth through the scaffolds’ pores was observed, providing notable support to the host bone. In quantitative terms, micro-CT and histomorphometry analysis post-implantation revealed no significant differences in bone regeneration across all groups. ConclusionThe 3D-printed scaffolds with perpendicular patterning, open porosity, and proposed composition displayed satisfactory mechanical properties, biocompatibility, and osteoconductive response. The scaffolds promoted bone regeneration at similar levels as the PLA. The highest contribution of bioCaP promoted a positive influence in certain histomorphometric parameters; however, it did not significantly improve their osteogenic capability. Further research is required to optimize scaffold composition and enhance their osteogenic potential. Clinical RelevanceThis study presents a significant advancement in bone tissue engineering through the development of personalized composite scaffolds for bone-related applications. The clinical implications of this research are profound, especially considering the increasing demand for functional bone regeneration technologies capable of producing cost-effective producing cost-effective customized scaffolds.

Isoginkgetin Inhibits RANKL-induced Osteoclastogenesis and Alleviates Bone Loss

Osteoporosis is characterized by excessive osteoclast activity leading to bone loss, decreased bone mineral density, and increased susceptibility to fractures. Through in vivo/vitro experiments, along with network pharmacology analysis, we aimed to explore the underlying mechanisms of Isoginkgetin (IGG) in inhibiting osteoclastogenesis, providing valuable insights for further research in the future. Firstly, we ascertained the safe concentration of IGG stimulation on BMMs, followed by a systematic exploration of the concentration gradient at which IGG inhibited osteoclastogenesis using TRAP analysis. An osteoporosis model was established to further validate the in vitro experimental findings by combining Micro-CT and immunohistochemical analysis. The results show that IGG did not exhibit cytotoxicity or proliferative effects on BMMs at concentrations equal to or less than 10 μM. Additionally, IGG inhibited the activity of osteoclastogenesis and bone resorption function at lower concentrations. RT-PCR and Western Blot results demonstrated that IGG could downregulate genes and proteins associated with osteoclastogenesis. The Western Blot results also showed that IGG inhibited the phosphorylation expression of P38, ERK, and P65 in the MAPK and NF-κB pathways. At the same time, it rescued the degradation of IκB-α at 15 and 60 min. IGG can also impact the relative expression levels of oxidative proteins such as SOD-1, HO-1, and catalase, thereby influencing cellular equilibrium and stress levels, ultimately inhibiting the formation of mature OC. In vivo experiments demonstrated that IGG alleviated bone loss caused by osteoclasts and improved relevant parameters of trabecular bone. So, IGG effectively attenuated osteoclastogenesis, and improved bone density, thereby portraying its role in osteoporosis management.

Microstructural Evaluation of Dental Implant Success Using Micro-CT: A Comprehensive Review

Micro-computed tomography (micro-CT) is an invaluable tool for the evaluation of dental implant success, whereby the assessment of bone microstructure is conducted. This review examines the role of micro-CT in evaluating bone microstructure in dental implants. A review of the current literature reveals that micro-CT enables the accurate measurement of bone volume, trabecular morphology, and connectivity density, all of which play a crucial role in implant stability. The high-resolution three-dimensional visualization capabilities of micro-CT are also beneficial for the analysis of osseointegration and the evaluation of bone augmentation biomaterials. Despite the existence of challenges such as imaging artifacts and limitations in in vivo applications, advancements in sub-micron resolution and artificial intelligence integration offer promise for improving diagnostic capabilities. Micro-CT provides valuable insights into bone microarchitecture and osseointegration dynamics, which have the potential to enhance pre-operative planning and clinical outcomes in dental implantology. Future research should prioritize the standardization of micro-CT analysis protocols and the exploration of direct clinical applications of this technology.

Comparison of Biocompatibility of 3D-Printed Ceramic and Titanium in Micropig Ankle Hemiarthroplasty.

Ankle arthritis is a common degenerative disease that progresses as cartilage damage in the lower tibia and upper talus progresses, resulting in loss of joint function. In addition to typical arthritis, there is also structural bone loss in the talus due to diseases such as talar avascular necrosis. Total talus replacement surgery is the procedure of choice in end-stage ankle arthritis and consists of a tibial, talar component and an insert. However, in cases of severe cartilage and bone damage to the talar bone with less damage to the tibial cartilage, a talar component hemiarthroplasty may be considered. Although the application of total talus replacement surgery using ceramics has been studied, reports on the application of metal 3D printing technology are limited. We aimed to investigate the feasibility of partial talar components using ceramic and titanium 3D printing technology in terms of biocompatibility and stability through animal experiments. Preoperative 3D CT was acquired and converted to STL files to fabricate a partial talus component for ankle hemiarthroplasty using ceramic and titanium. Six minipigs with an average age of 17 months were implanted with three ceramic (C-group) and three titanium talar components (T-group) in the hind limb ankle joint. The surgery was performed under anesthesia in a sterile operating room and was performed by two experienced foot and ankle specialist orthopedic surgeons. Blood analysis and CT were performed before surgery and every month for 3 months after surgery to assess the extent of inflammatory response and physical stability, sacrifices were performed 3 months after surgery, and H&E staining and micro-CT analysis were performed to compare histological biocompatibility. A grading score was calculated to semi-quantitative assess and compare the two groups. In the postsurgical evaluation, blood analysis revealed that both groups had increased white blood cell counts on the postoperative day after surgery. The white blood cell count increased more in the titanium group (1.85-fold) than in the ceramic group (1.45-fold). After 3 months, all values normalized. During the study, CT analysis confirmed that all artificial samples were displaced from their initial positions. In micro-CT analysis, the adhesive tissue score of the ceramic artificial sample was better than that of the titanium sample (average threshold = 3027.18 ± 405.92). In histologic and grading scores for the inflammatory reactions, the average inflammation indices of the ceramic and titanium groups were 2.0 and 1.21, respectively. Also, the average grade score confirmed based on the results of fibrous tissue proliferation and new blood vessels was 18.4 in the ceramic application group and 12.3 in the titanium application group. In conclusion, both titanium and ceramics have excellent biocompatibility for artificial joints, and ceramic materials can be used as novel artificial joints. Further research on the strength and availability of these ceramics is required.

Regeneration of Chronic Alveolar Vertical Defects Using a Micro Dosage of rhBMP-2. An Experimental InVivo Study.

The objective of this study is to compare the effect of the location of recombinant human bone morphogenetic protein 2 (rhBMP-2) from the native bone and the periosteum for vertical alveolar bone augmentation. Mandibular, chronic, standardized, bilateral, and vertical defects in 12 beagle dogs were evaluated using four modalities: a xenograft alone (XENO; n = 6); rhBMP-2 alone (BMP; n = 6); a technique with rhBMP-2 close to the host bone covered by xenograft (SAN; n = 6); and a technique with rhBMP-2 close to the flap on top of the xenograft (LAS; n = 6). After 8 weeks, a series of invivo inspections, fluorescence microscopy, histologic and histomorphometric evaluations, and micro-CT analyses. After 8 weeks of healing, new bone formation correlated with proximity of rhBMP to the perforated membrane with BMP and LAS (p = 0.024). The highest total bone volume was found in the LAS group (45.1% ± 13.3%), followed by the SAN group (35.2% ± 6.7%), BMP group (33.1% ± 11.8%), followed by the XENO group (23.1% ± 6.5%). The SAN group demonstrated frequent seroma formation. Blood vessel formation was more pronounced in the LAS + rhBMP group, with a significant increase of 27.1% compared to the XENO group (p = 0.02). Micro-CT revealed a strong trend for higher bone volume in the BMP group (34.7%) compared to the XENO group (13.6%) (p = 0.06). Only rhBMP-2 groups demonstrated bone formation above the perforated membrane. The location of rhBMP-2 in relation to the biomaterial and periosteum influenced the effectiveness of vertical bone regeneration.

Effects of Polydeoxyribonucleotide (PDRN) on Endosinus Bone Regeneration Following Sinus Floor Elevation: An Experimental InVivo Pilot Study.

To determine the effect of polydeoxyribonucleotide (PDRN) on endosinus bone regeneration in a rabbit sinus model at sequential healing time points. Eighteen New Zealand white rabbits were used. Bilateral sinus floor elevation (SFE) was performed. Two groups were randomly assigned to each sinus: (1) test group, in which SFE was performed using collagenated bone substitute material soaked with PDRN (concentration 2.0 mg/mL, dose 0.5 mL), and (2) control group, in which SFE was performed using collagenated bone substitute material only. The experimental animals were sacrificed at 2, 4, and 8 weeks (n = 6 at each healing time point). Microcomputed tomography (micro-CT), histologic, and histomorphometric analyses were performed. The micro-CT analysis revealed statistically significant increases in the mineralized tissue volume between 4 and 8 weeks (p < 0.05). Histologically, no specific intergroup difference was found in the pattern of new bone formation. Histomorphometrically, the area of newly formed bone (NB) was larger in the test group than in the control group at all healing time points (1.4 vs. 1.2 mm2 at 2 weeks, 3.4 vs. 1.9 mm2 at 4 weeks, and 5.7 vs. 4.5 mm2 at 8 weeks; median value), but the difference was statistically significant only at 4 weeks (p < 0.05). NB in set regions of interest (ROI_C, ROI_W, and ROI_M) tended to be greater in the test group than in the control group without statistical significance (p > 0.05). PDRN appeared to enhance new bone formation at all healing time points, but the improvement was statistically significant only at 4 weeks.

Multilayered Cryogel Enriched with Exosomes Regenerates and Maintains Cartilage Architecture and Phenotype in Goat Osteochondral Injuries.

Treatment of critical-size osteochondral (OC) injuries at load-bearing sites has remained a major clinical challenge in orthopedic surgery. This is due to the anisotropic characteristics of OC tissue and the stratified structure of the cartilage. Here, we developed a multilayered OC scaffold by employing cryogelation technology. Gelatin, chitosan, and chondroitin sulfate were utilized for designing three distinct, 2425 ± 120 μm thick layers of cartilage having different alignments, while nanohydroxyapatite and gelatin were used for the subchondral bone layer. Exosomes derived from articular chondrocytes in the range of 60-110 nm were used to promote chondrogenesis. The biocompatibility and cartilage formation potential of the scaffold and exosomes were initially evaluated in rat OC defects. The application of exosome-loaded scaffolds was then investigated in a critical-size OC injury (8 × 10 mm) created in the goat knee. Artificial synovial fluid was designed and utilized as a carrier for exosomes for a booster dose administered as an intra-articular injection. X-ray imaging and micro-CT analysis revealed that the treatment resulted in improved subchondral bone regeneration. The defect region exhibited healthy hyaline cartilage formation, as detected by MRI imaging. Moreover, histological examination revealed that the treatment group showed augmented cell proliferation, matrix deposition, secretion of proteoglycans, and the formation of stratified hyaline cartilage over a long-term (6 and 12 months), whereas the control group demonstrated the formation of fibrocartilage. Treatment-induced upregulation of collagen II, aggrecan, and SOX 9 genes (∼10 fold) further provided evidence that the cartilage phenotype was well preserved. Hence, the proposed treatment has significant translational potential for treating adverse OC clinical injuries.

In Vivo Study of Organ and Tissue Stability According to the Types of Bioresorbable Bone Screws.

Biodegradable material, such as magnesium alloy or polylactic acid (PLA), is a promising candidate for orthopedic surgery. The alloying of metals and the addition of rare earths to increase mechanical strength are still questionable in terms of biosafety as absorbent materials. Therefore, the purpose of this study is to understand the effect of substances due to the degradation of various biodegradable substances on organs in the body or surrounding tissues. A total of eighty male Sprague-Dawley rats were selected for this study, and the animals were divided into four groups. Each of the three experimental groups was implanted with magnesium alloy, polymer, and titanium implants; the control group only drilled into the cortical bone. Serum assay, micro-CT, hematoxylin and eosin staining, immunoblotting, and real-time PCR were evaluated. There was no significant difference between the two groups of magnesium alloy and polymer in serum assay, but micro-CT analysis confirmed that magnesium alloy degrades faster than polymer, and histological examination showed a strong inflammatory response in the early stages, which was similarly observed in immunoblotting and real-time PCR. Our findings show that there was no toxicity due to the degradation of the biodegradable material, and the difference in each inflammatory response is thought to be determined by the rate of degradation in the body.

TiNbSn alloy plates with low Young's modulus modulates interfragmentary movement and promote osteosynthesis in rat femur

Orthopedic implants such as arthroplasty prostheses, fracture plates, and intramedullary nails often use materials like Ti6Al4V alloy and commercially pure titanium (CP-Ti), which have Young's modulus significantly higher than that of human cortical bone, potentially causing stress shielding and inhibiting effective fracture healing. TiNbSn alloy, a β-type titanium alloy with a lower Young's modulus (40–49 GPa), has shown promise in reducing stress shielding and enhancing bone healing by promoting effective load sharing with bone. This study used 5-hole plates made from TiNbSn alloy and CP-Ti to investigate their effects on bone healing in a rat femoral fracture model. Micro-CT analysis and mechanical testing were performed six weeks postoperatively to assess bone healing. Additionally, Finite element method (FEM) analysis was employed to evaluate stress shielding and interfragmentary movement (IFM) at the fracture site. Micro-CT analysis revealed significantly higher bone volume and mineral density in the TiNbSn group than in the CP-Ti group. Mechanical testing showed increased maximum load and stiffness in the TiNbSn group (77.2 ± 10.0 N for the TiNbSn alloy plate group versus 53.3 ± 8.5 N for the CP-Ti group (p = 0.002)). FEM analysis indicated that TiNbSn plates reduced stress shielding and allowed for greater displacement and strain, promoting IFM conducive to bone healing. The findings suggest that TiNbSn alloy plates are more effective than CP-Ti plates in promoting bone healing by reducing stress shielding and enhancing IFM. The lower Young's modulus of TiNbSn allows better load distribution, facilitating bone regeneration and strengthening at the fracture site.

Prenatal Exposure to a Human Relevant Mixture of Endocrine-Disrupting Chemicals Affects Mandibular Development in Mice.

Mandible is a bony structure of neuroectodermal origin with unique characteristics that support dentition and jaw movements. In the present study, we investigated the effects of gestational exposure to a mixture of endocrine-disrupting chemicals (EDCs) on mandibular growth in mice. The mixture under study (Mixture N1) has been associated with neurodevelopmental effects in both a human cohort and animal studies. Pregnant mice were exposed throughout gestation to 0.5× (times of pregnant women's exposure levels), 10×, 100× and 500× of Mixture N1, or the vehicle, and the mandibles of the male offspring were studied in adulthood. Micro-CT analysis showed non-monotonic effects of Mixture N1 in the distances between specific mandibular landmarks and in the crown width of M1 molar, as well as changes in the mandibular bone characteristics. The alveolar bone volume was reduced, and the trabecular separation was increased in the 500× exposed mice. Bone volume in the condyle head was increased in all treated groups. Τhe Safranin-O-stained area of mature hypertrophic chondrocytes and the width of their zones were reduced in 0.5×, 10× and 100× exposed groups. This is the first indication that prenatal exposure to an epidemiologically defined EDC mixture, associated with neurodevelopmental impacts, can also affect mandibular growth in mammals.

Revisiting the scorpion central nervous system using microCT

The central nervous system (CNS) of Chelicerata has remained conserved since the Cambrian, yet few studies have examined its variability within chelicerate orders including Scorpiones. The scorpion CNS comprises the prosomal ganglion and opisthosomal ventral nerve cord. We visualize the scorpion CNS with microCT, explore morphological variation across taxa, compare the scorpion CNS to other arachnids, and create a terminology glossary and literature review to assist future studies. Six scorpion species were microCT scanned. Scan quality varied and most structures in the prosomal ganglion could only be observed in Paruroctonus becki (Vaejovidae). Major nerves and the first opisthosomal ganglion were visible in nearly all taxa. We present the most detailed 3D-rendering of the scorpion prosomal ganglion to date. Our results corroborate existing research and find the scorpion CNS to be conserved. Nearly all structures reported previously in the prosomal ganglion were located in similar positions in P. becki, and nerve morphology was conserved across examined families. Despite similarities, we report differences from the literature, observe taxonomic variation in prosomal ganglion shape, and confirm positional variation for the first opisthosomal ganglion. This study serves as a starting point for microCT analysis of the scorpion CNS, and future work should include more distantly related, size variable taxa to better elucidate these findings.

Micro-CT Analysis of Pore Structure in Upland Red Soil Under Different Long-Term Fertilization Regimes

This study hypothesized that long-term fertilization alters the pore structure and aggregate stability in upland red soil. A long-term fertilization experiment in Qiyang, Hunan, was conducted with three treatments: no fertilization (CK), nitrogen–phosphorus–potassium fertilization (NPK), and nitrogen–phosphorus–potassium combined with pig manure (NPKOM). X-ray computed tomography (CT) scanning technology was used to assess three-dimensional pore structures at both the soil column (50 mm diameter and 50 mm height) and aggregate scales (diameter 3–5 mm), alongside the evaluation of the soil’s physical and chemical properties. Results showed that the soil organic carbon content (SOC) increased by 44.8% in NPK and 112.5% in NPKOM compared to CK. NPKOM improved the aggregate stability by 51.6%, whereas NPK had no significant effect. At the soil column scale, NPK increased the total porosity by 13.7% but reduced larger pores (&gt;0.06 mm), whereas NPKOM decreased the total porosity by 7.8% and increased larger pores. At the aggregate scale, NPKOM increased the porosity for pores &gt;0.098 mm by 7.6 times compared to CK and 9.5 times compared to NPK. In conclusion, long-term NPKOM significantly enhances the SOC and aggregate stability and promotes larger pore formation, unlike NPK, which mainly increases SOC but does not improve the soil structure.

Sex hormone deficiency in male and female mice expressing the Alzheimer's disease-associated risk-factor TREM2 R47H variant impacts the musculoskeletal system in a sex- and genotype-dependent manner.

The R47H variant of the triggering receptor expressed on myeloid cells 2 (TREM2) is a risk factor for Alzheimer's disease in humans and leads to lower bone mass accrual in female but not male 12-mo-old mice. To determine whether, as with aging, gonadectomy results in sex-specific musculoskeletal effects, gonad removal or SHAM surgery was performed in 4-mo-old TREM2R47H/+ mice and WT male and female littermates (n = 10-12/group), with sexes analyzed separately. Body weight was lower in males, but higher in females after gonadectomy, independently of their genotype. Gonadectomy also leads to decreased BMD in males at all sites and in the whole body (total) and spine in female mice for both genotypes. Total and femur BMD was lower in gonadectomized male mice 6-wk post-surgery, independently of the genotype. On the other hand, BMD was only lower in ovariectomized WT but not TREM2R47H/+ mice in all sites measured at this time point. Bone formation and resorption marker levels were not affected by orchiectomy, whereas CTX was higher 3wk after surgery and P1NP showed a tendency toward lower values at the 6-wk time point only in ovariectomized WT mice. Micro-CT analyses showed no differences resulting from gonadectomy in structural parameters in femoral cortical bone for either sex, but lower tissue mineral density in males of either genotype 6-wk post-surgery. Nevertheless, biomechanical properties were overall lower in gonadectomized males of either genotype, and only for WT ovariectomized mice. Distal femur cancellous bone structure was also affected by gonadectomy in a genotype- and sex-dependent manner, with genotype-independent changes in males, and only in WT female mice. Thus, expression of the TREM2 R47H variant minimally alters the impact of gonadectomy in the musculoskeletal system in males, whereas it partially ameliorates the consequences of ovariectomy in female mice.

Histomorphometric and Micro-CT Evaluation of Cerabone and Bio-Oss in Maxillary Sinus Lifting: A Randomized Clinical Trial.

Background and Objectives: The loss of teeth in the posterior maxillary region often leads to significant alveolar bone resorption and maxillary sinus pneumatization, complicating dental implant placement. Maxillary sinus grafting, typically using autogenous bone, is a common solution. However, autogenous bone grafts require additional surgical procedures, leading to increased morbidity. This study aims to compare the efficacy of two xenografts, Bio-Oss and Cerabone, in promoting new bone formation in maxillary sinus grafting through histomorphometric analysis and micro-computed tomography (micro-CT). Materials and Methods: A total of 22 maxillary sinuses (12 right and 10 left) were grafted, with 12 using Cerabone and 10 using Bio-Oss. Six months post-grafting, biopsies were collected for histomorphometric analysis to measure new bone formation, connective tissue, and residual biomaterial. Additionally, micro-CT analysis was performed to assess bone volume fraction, trabecular thickness, number, and separation. Results: Histomorphometric analysis showed that the Cerabone group had a higher average new bone formation (25.94% ± 10.55) compared to the Bio-Oss group (17.29% ± 4.61), with a statistically significant difference (p = 0.02). Micro-CT analysis revealed that the bone volume fraction in the Cerabone group was significantly higher compared to the Bio-Oss group, with significant differences in trabecular thickness (p = 0.02) but not in trabecular number or separation. Conclusions: The study demonstrates that both xenografts are effective in promoting new bone formation in maxillary sinus grafting. However, Cerabone showed superior performance in terms of new bone formation and bone volume fraction, suggesting it may be a more effective option for maxillary sinus augmentation procedures.

Abnormally accumulated GM2 ganglioside contributes to skeletal deformity in Tay-Sachs mice.

Tay-Sachs Disease is a rare lysosomal storage disorder caused by mutations in the HEXA gene, responsible for the degradation of ganglioside GM2. In addition to progressive neurodegeneration, Tay-Sachs patients display bone anomalies, including kyphosis. Tay-Sachs disease mouse model (Hexa-/-Neu3-/-) shows both neuropathological and clinical abnormalities of the infantile-onset disease phenotype. In this study, we investigated the effects of GM2 accumulation on bone remodeling activity. Here, we evaluated the bone phenotype of 5-month-old Hexa-/-Neu3-/- mice with age-matched control groups using gene expression analysis, bone plasma biomarker analysis, and micro-computed tomography. We demonstrated lower plasma alkaline phosphatase activity and calcium levels with increased tartrate-resistant acid phosphatase levels, indicating reduced bone remodeling activity in mice. Consistently, gene expression analysis confirmed osteoblast reduction and osteoclast induction in the femur of mice. Micro-computed tomography and analysis show reduced trabecular bone volume, mineral density, number, and thickness in Hexa-/-Neu3-/- mice. In conclusion, we demonstrated that abnormal GM2 ganglioside accumulation significantly triggers skeletal abnormality in Tay-Sachs mice. We suggest that further investigation of the molecular basis of bone structure anomalies is necessary to elucidate new therapeutic targets that preventthe progression ofbone symptoms and improve the life standards of Tay-Sachs patients. KEY MESSAGES: We detected the markers of bone loss-associated disorders such as osteopenia and osteoporosis in the Tay-Sachs disease mice model Hexa-/-Neu3-/-. We also demonstrated for the first time there is an increase in trabecular spacing and a reduction in trabecular thickness and number indicating skeletal abnormalities in mice model using micro-CT analysis.

Adipose-derived stem cell exosomal miR-21-5p enhances angiogenesis in endothelial progenitor cells to promote bone repair via the NOTCH1/DLL4/VEGFA signaling pathway

BackgroundAngiogenesis is essential for repairing critical-sized bone defects. Although adipose-derived stem cell (ADSC)-derived exosomes have been shown to enhance the angiogenesis of endothelial progenitor cells (EPCs), the underlying mechanisms remain unclear. This study aims to explore the effects and mechanisms of ADSC-derived exosomes in enhancing bone repair by promoting EPC angiogenesis.MethodsTransmission electron microscopy, nanoparticle tracking analysis, and Dil reagent kit were employed to identify ADSC-derived exosomes and their internalization by EPCs. Micro-CT analysis, H&E staining, and Masson staining were used to assess bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N), as well as the pathological changes and fibrosis at defect sites. Cell viability, migration, invasion, and tube formation of EPCs were evaluated using CCK-8, wound healing, Transwell, and tube formation assays. Immunohistochemical staining, RT-PCR, and Western blotting were utilized to measure the gene and protein expression of markers such as CD31, VEGFA, OCN, RUNX2, NOTCH1, and DLL4. Gene sequencing and bioinformatics analyses were conducted to identify the most highly expressed miRNA in exosomes, while miRDB and dual-luciferase reporter assays were used to explore the interaction between miR-21-5p and NOTCH1.ResultsThe ADSC-derived exosomes, averaging 126 nm in diameter, were internalized by EPCs. In vivo, these exosomes promoted new bone formation, increased BMD, BV/TV, Tb.Th, and Tb.N, reduced pathological damage to cranial defect tissues, enhanced vascular and bone tissue regeneration, and upregulated OCN and RUNX2 expression. In vitro, ADSC-derived exosomes enhanced EPC viability, migration, invasion, and tube formation. Both in vivo and in vitro experiments demonstrated that ADSC-derived exosomes upregulated CD31 and VEGFA expression. miR-21-5p, the most highly expressed miRNA in ADSC-derived exosomes, was found to target NOTCH1. Overexpression of miR-21-5p in these exosomes facilitated EPC migration, tube formation, and VEGFA expression while downregulating NOTCH1 and DLL4 expression. Inhibition of miR-21-5p produced opposite effects on EPCs.ConclusionsThese findings indicate that miR-21-5p in ADSC-derived exosomes promotes angiogenesis in EPCs to accelerate bone repair by targeting the NOTCH1/DLL4/VEGFA signaling pathway, offering a potential therapeutic strategy for bone defect treatment.Graphical

Microarchitectural Study of the Augmented Bone Following a Modified Ridge Splitting Technique: Histological and Micro-Computed Tomography Analyses.

Objectives: The aim of this matched prospective cohort study was to examine the microarchitecture of the augmented bone following a modified alveolar ridge splitting procedure and compare it to that of native bone. Methods: In the test group, patients underwent a modified ridge split osteotomy procedure to restore the width of the posterior segment of the mandible. Patients with sufficient bone width for dental implant placement in the posterior region of the mandible following 3-month-long spontaneous healing after tooth removal were included in the control group. In both study groups, bone biopsy samples were harvested and dental implants were placed. Histomorphometry and micro-CT analysis were performed. Results: Altogether, 15 patients were included in this study (7 patients in the test group, with 14 bone core biopsies harvested, and 8 patients in the control group, with 13 bone core biopsies harvested). Percentage bone volume (BV/TV) in the micro-CT analysis (22.088 ± 8.094% and 12.075 ± 4.009% for the test and control group, respectively) showed statistically significant differences between study groups. Conclusions: Based on histological and micro-CT analyses, the modified ridge splitting procedure with autologous bone block harvested from the retromolar area results in a dental implant recipient bone microarchitecture superior to that of the extraction sockets left to heal undisturbed for a 3-month-long healing period.

Growth of a Tessellation: Geometric rules for the Development of Stingray Skeletal Patterns.

The skeletons of sharks and rays, fashioned from cartilage, and armored by a veneer of mineralized tiles (tesserae) present a mathematical challenge: How can the continuous covering be maintained as the skeleton expands? This study, using microCT and custom visual data analyses of growing stingray skeletons, systematically examines tessellation patterns and morphologies of the many thousand interacting tesserae covering the hyomandibula (a skeletal element critical to feeding), over a two-fold developmental change in hyomandibula length. The number of tesserae remains surprisingly constant, even as the hyomandibula expands isometrically, with all hyomandibulae displaying self-similar distributions of tesserae shapes/sizes. Although the distribution of tesserae geometries largely agrees with the rules for polyhedra tiling of complex surfaces-dominated by hexagons and a minor fraction of pentagons and heptagons, but very few other polygons-the agreement with Euler's classic mathematical laws is not perfect. Contrary to the assumed uniform growth rate (which is shown would create geometric incompatibilities), larger tesserae grow faster to accommodate skeletal expansion. It is hypothesized that this local regulation of global system complexity is driven by tension (from cartilaginous core expansion) in the fibers connecting tesserae, with strain-responsive cells orchestrating local mineral apposition.

Fabrication of hierarchically porous trabecular bone replicas via 3D printing with high internal phase emulsions (HIPEs)

Combining emulsion templating with additive manufacturing enables the production of inherently porous scaffolds with multiscale porosity. This approach incorporates interconnected porous materials, providing a structure that supports cell ingrowth. However, 3D printing hierarchical porous structures that combine semi-micropores and micropores remains a challenging task. Previous studies have demonstrated that using a carefully adjusted combination of light absorbers and photoinitiators in the resin can produce open surface porosity, sponge-like internal structures, and a printing resolution of about 150µm. In this study, we explored how varying concentrations of tartrazine (0, 0.02, 0.04, and 0.08 wt%) as a light absorber affect the porous structure of acrylate-based polymerized medium internal phase emulsions fabricated via vat photopolymerization. Given the importance of a porous and interconnected structure for tissue engineering and regenerative medicine, we tested cell behavior on these 3D-printed disk samples using MG-63 cells, examining metabolic activity, adhesion, and morphology. The 0.08 wt% tartrazine-containing 3D-printed sample (008 T) demonstrated the best cell proliferation and adhesion. To show that this high internal phase emulsion (HIPE) resin can be used to create complex structures for biomedical applications, we 3D-printed trabecular bone structures based on microCT imaging. These structures were further evaluated for cell behavior and migration, followed by microCT analysis after 60 days of cell culture. This research demonstrates that HIPEs can be used as a resin to print trabecular bone mimics using additive manufacturing, which could be further developed for lab-on-a-chip models of healthy and diseased bone.