Search Results (274)

Background/Objectives: Age estimation is of fundamental importance in forensic investigations. When traditional methods based on gross bone morphology or morphometric analysis cannot be applied, forensic experts must rely on multidisciplinary approaches. Histomorphometry has consistently proven to be reliable in cases of highly fragmented or incomplete skeletal remains, particularly in older individuals. Building on the foundational study of Amprino and Bairati, this study evaluated the correlations between bone microstructural features in femoral cross-sections and the age and sex of individuals. Methods: The sample comprised 95 femoral mid-diaphyseal thin sections obtained from autopsy specimens housed at the Institute of Legal Medicine, University of Bari (Italy), representing both male and female individuals aged 18 to 92 years. The numbers and densities of primary, intact secondary, and fragmentary secondary osteons, together with osteon circularity and the mean osteonal area, were measured to investigate age-related variation. Statistical analyses included t-tests, Mann–Whitney tests, Spearman’s rank correlation, and General Linear Models (GLMs). Results: No significant differences in histomorphometric variables were observed between males and females. However, the number of intact secondary osteons and osteon population density increased with age, while the mean osteonal area and osteon circularity decreased with age. Although some variables displayed significant correlations with age, residual analysis indicated a lack of heterogeneity in variance, which limited the development of a robust predictive model. Conclusions: The findings highlight both the potential and the limitations of histomorphometry in forensic age estimation. While certain microstructural variables correlate with age, inter-individual variability reduces predictive accuracy. Further research is needed to refine models that account for biological and biomechanical variability, particularly in older adults. Full article

Background: Traditional parameters such as bone-to-implant contact percentage (BIC%) provide only static insights into implant integration and do not reflect the temporal dynamics of bone regeneration. The concept of Dynamic Regenerative Balance (DRB) was introduced to represent the biological equilibrium between bone formation and graft resorption. The break-even point serves as a measurable approximation of this equilibrium. This study aimed to illustrate the usefulness of the break-even point in expressing the balance between graft resorption and new bone formation, rather than to define definitive values for specific biomaterials. Methods: Four preclinical studies on sinus floor elevation in rabbits were selected. Each reported histomorphometric data on new bone formation and graft resorption at two or more time points. Six biomaterials were analyzed: autogenous bone, Bio-Oss^®, Bio-Oss Collagen^®, Gen-Os^®, Maxresorb^®, and Maxresorb^® Inject. The break-even point was calculated by linear extrapolation as the time at which new bone equals residual graft percentage. Results: The break-even point varied significantly among biomaterials (expressed in days/area %): autogenous bone reached equilibrium fastest (18.4 days/13.5%), followed by Gen-Os^® (40.4 d/19.1%). Bio-Oss Collagen^® (62.3 d/28.3%), Maxresorb^® (73.9 d/36.4%), and Maxresorb^® Inject (96.1 d/34.1%). For Bio-Oss^®, it occurred at 81.8 days (33.6%) in one study, while in another, it was not reached within 6 months. These differences reflect distinct regenerative kinetics and resorption profiles among materials. Conclusions: The break-even point offers a simple and informative parameter to describe the balance between graft resorption and new bone formation, providing a useful complement to conventional histomorphometric measures and a framework for future studies. Full article

Orthopedic implants have a critical role in modern medical practice, being useful in bone regeneration, joint arthroplasty, and healing fractures. The success of osseointegration depends on implant properties (composition, stability, geometry, biocompatibility) and host factors (local reactivity, comorbidities). Preclinical evaluation in animal models is essential before clinical application. In orthopedic implantology, the selection and real utility of a range of animals are important, with an emphasis placed on bone–implant interface, biomechanical function, and long-term integration. Smaller animals such as rabbits and rats have widespread use in early biocompatibility and osseointegration testing, but larger animals such as pigs, sheep, and canines have a larger physiological bone similarity and can, therefore, be utilized for bearing loads in testing. Considering the utility and disadvantages of certain species—including suitability for new biomaterials, coatings, and biomechanical function—this article discusses testing methodologies such as push-out/pull-out tests, histomorphometry, and micro-CT and their utility in testing the integration of implants and regeneration of bone. Conclusions confirm a multi-species model in use in preclinical testing for the development of implants and improvements in clinical success. Unlike previous reviews, this article emphasizes translational strategies, integrates ethical perspectives in model selection, and discusses the synergistic use of imaging modalities with biomechanical tests for comprehensive assessment. Full article

Shilajit, a natural herbo-mineral compound with potent antioxidant, anti-inflammatory, and osteogenic properties, has been traditionally used to promote tissue repair. However, limited experimental data exist on its localized application in bone regeneration. This study aimed to evaluate the combined effect of Shilajit and bovine-derived xenograft on bone healing in a rat tibial defect model. Twenty-eight male Sprague–Dawley rats were randomly assigned to four groups (n = 7): Control (defect left to heal spontaneously), Graft-only, Graft + Shilajit 150 mg/kg, and Graft + Shilajit 250 mg/kg. Standardized 3 mm tibial defects were created and filled with xenograft in all groups except the Control. Shilajit was administered intraperitoneally on days 0–3 postoperatively. After 4 weeks, serum total oxidant status (TOS), total antioxidant status (TAS), and TNF-α levels were measured. Tibial specimens underwent histopathological, histomorphometric, and TNF-α immunohistochemical analysis. High-dose Shilajit significantly increased TAS and reduced TOS compared with the Control and Graft-only groups (p < 0.001). Median TNF-α concentrations decreased in a dose-dependent manner, with the lowest values in the high-dose group (15.7 [14.3–17.1] pg/mL, p < 0.001). Histomorphometry revealed the highest new bone area percentage (78.1% [74.9–81.2]) and lowest fibrous tissue content (9.8% [8.1–11.6]) in the high-dose group. Immunohistochemistry confirmed marked suppression of TNF-α expression in Shilajit-treated groups, particularly at high doses. The combination of Shilajit and bovine-derived xenograft significantly enhanced bone regeneration in a dose-dependent manner, likely through antioxidative, anti-inflammatory, and osteogenic mechanisms. These findings suggest that Shilajit may serve as a promising adjunct in bone grafting procedures. Full article

Background: Metabolic stress from amino acid (AA) insufficiency is increasingly linked to pathological angiogenesis, but specific essential AA (EAA) roles remain undefined. Neovascular age-related macular degeneration (AMD), a major cause of blindness driven by aberrant ocular neovascularization, has limited efficacy with current VEGFA-targeting therapies. We sought to identify specific EAAs that regulate pathological angiogenesis and dissect their mechanisms to propose new therapeutic strategies. Methods: Human retinal microvascular endothelial cells (HRMVECs) were used to identify angiogenesis-regulating amino acids through systematic EAA screening. The molecular mechanism was investigated using shRNA-mediated knockdown of key stress response regulators (HRI, PKR, PERK, GCN2) and ATF4. Angiogenesis was assessed via tubule formation and migration assays. Therapeutic potential was examined in a laser-induced choroidal neovascularization (CNV) mouse model, evaluated by fluorescein angiography and histomorphometry. Results: Deprivation of methionine, lysine, and threonine potently induced capillary-like tube formation (p < 0.01). Mechanistically, restriction of these three EAAs activated HRI and GCN2 kinases, converging on eIF2α phosphorylation to induce ATF4 and its target VEGFA. Dual, but not single, knockdown of HRI and GCN2 abolished eIF2α-ATF4 signaling and angiogenic responses. Restricting these EAAs exacerbated CNV area in mice. Conclusions: Our findings reveal a coordinated HRI/GCN2-ATF4-VEGFA axis linking EAA scarcity to vascular remodeling, establishing proof-of-concept for targeting this pathway in CNV. This work highlights the therapeutic potential of modulating specific AA availability or targeting the HRI/GCN2-ATF4 axis to treat CNV. Full article

Background: The aim of this experimental pilot study was to evaluate the effect of pore volume and material composition on bone ingrowth into a resorbable poly-L-lactide-CaCO₃/CaP scaffold. Methods: Cylindric scaffolds of 7 mm diameter and 5 mm height and two different degrees of porosity were produced using selective laser sintering of poly-L-lactide-powder containing 24% CaCO₃ spherulites with and without surface modification with 4% CaP. Six minipigs received the four types of macroporous cylindrical scaffolds, inserted press fit into trephine defects of the tibial metaphyses, and left to heal for 4 and 13 weeks in three animals each. The specimens were evaluated using µCT for pore volume fill, and histomorphometry for bone formation and immunohistochemistry for expression of osteocalcin. Results: After 4 weeks, newly formed bone ranged from 2.73 mm² to 5.28 mm² mean total area. Mean pore volume fill varied between 12.25% and 20.35% and the average level of osteocalcin expression ranged from 2.49 mm² to 4.48 mm² mean total area. No significant differences were found between the different scaffolds. After 13 weeks, bone formation and pore fill volume had significantly increased in all scaffold groups up to a mean value of 14.79 mm² and 96.04%, respectively. Again, differences between the groups were not significant. Conclusions: The tested SLS produced scaffolds allowed for bone ingrowth, almost completely filling the pore volume after 13 weeks. Newly formed bone was in direct contact with the scaffold walls. Differences in pore volume did not account for significant differences in bone formation inside the scaffolds. The addition of CaP likewise did not lead to increased bone formation, most likely due to low availability of CaP to the biological environment. Full article

This study focused on the effects of including a mixture of Hermetia illucens (HI) and Tenebrio molitor (TM) meals on feed colour, growth performance, carcass yield, somatic indexes, nutrient digestibility, intestinal microbiota, histomorphometry, and fillet quality of rainbow trout (Oncorhynchus mykiss). Seven diets were formulated: one control diet (CTRL) containing 15% fishmeal, two diets where the fishmeal was replaced with 100% HI (HI100) or TM (TM100) meal, and four diets containing increasing levels of a mix (1:1) of HI and TM meals, replacing 25% (MIX25), 50% (MIX50), 75% (MIX75) and 100% (MIX100) of the fishmeal. A total of 378 fish (average weight: 126 ± 1.71 g (g)) were randomly distributed in 21 experimental tanks (3 replicates/treatment and 18 fish/replicate) and fed for 84 days. A digestibility trial was also conducted to assess the apparent nutrient digestibility coefficients of the diets. Diet significantly affected pellet colour (L*, a*, b*; p < 0.001), with CTRL and MIX25 being the lightest and HI100 and MIX75 the darkest. Fillet colour, pH_24h, and drip loss were significantly affected by diet (p < 0.001). The pH_24h parameter, reflecting stress and freshness, was lowest in MIX25 and highest in MIX75, suggesting a synergistic effect and potential impact on shelf life. No significant differences were found in growth performance, carcass yield, somatic indexes, fillet chemical composition, intestinal microbiota, and the histopathology of internal organs among the experimental diets (p > 0.05), nor in nutrient digestibility. This study confirms that HI and TM meals, alone or combined, are viable alternatives in low-fishmeal diets for rainbow trout. Full article

Background/Objectives: Fructooligosaccharides (FOS) and dried tart cherry (TC) are examples of simple and complex (i.e., within a food matrix) prebiotics that have demonstrated promising osteoprotective activity. In this study, we examined how dietary supplementation with TC or FOS shapes the gut-bone axis to promote bone accrual in young adult mice, and the role of the gut microbiota in mediating these responses. Methods: Studies were performed using 10-wk-old female C57BL/6 mice (n = 10–12/group) fed a control diet or control diet supplemented with 10% TC or FOS for 10 wks alone or in combination with an antibiotic/anti-fungal cocktail to suppress the gut microbiota. The bone phenotype was characterized by dual-energy X-ray absorptiometry, micro-computed tomography and static and dynamic bone histomorphometry. The gut-microbiota was profiled and short chain fatty acids (SCFA) were assessed based on 16S rRNA profiling and gas chromatographic techniques, respectively. Results: FOS and TC enhanced bone structure, with FOS yielding more pronounced benefits across cortical and trabecular compartments. These skeletal improvements with FOS occurred in the absence of systemic changes in bone turnover markers but were accompanied by increases in local bone formation, osteoblast and osteocyte numbers, and bone mineralization in the femur. Both diets altered gut microbiota composition and increased fecal concentrations of the most abundant SCFAs (i.e., acetate, propionate and butyrate), but the response was greater with FOS. Suppression of the gut microbiota and fecal SCFAs with the antibiotic/anti-fungal cocktail inhibited the effects of FOS and TC on cortical bone, but induced unexpected improvements in the trabecular bone. Conclusions: These findings demonstrate differential effects of simple and complex prebiotics on the gut-bone axis in young adult female mice and support a role for SCFA in the cortical bone response, but not in the trabecular bone response with this model of gut microbiota suppression. Full article

The effects of varying dietary metabolizable energy (ME) and crude protein (CP) levels, along with their interactive effects, on the apparent nutrient metabolism, development of digestive organs, intestinal morphology, and microbiota composition in Jingyuan chickens during the growing phase were evaluated. A total of 540 seven-week-old male Jingyuan chickens were randomly assigned to nine groups, with six replicates per group and 10 chickens per replicate. The trial lasted for 11 weeks. A 3 × 3 factorial design was adopted, comprising three levels of ME, namely, low (11.28 MJ/kg, LE group), medium (11.70 MJ/kg, ME group), and high (12.12 MJ/kg, HE group) and three levels of CP, namely, low (14.00%, LP group), medium (15.50%, MP group), and high (17.00%, HP group). The levels of ME and CP, along with their interactions, had significant effects on the average daily gain (ADG), average daily feed intake, feed conversion ratio (FCR), apparent metabolizable rate of CP, gizzard weight, duodenal lengths, jejunal villus height (VH), crypt depth (CD), and muscle layer thickness (MLT) (p < 0.05). The combination of medium level ME (11.70 MJ/kg) and medium level CP (15.50%) (MEMP group) exhibited the best performance, with the highest ADG and the lowest FCR (p < 0.05). Moreover, this group showed significantly higher duodenal length, jejunal CD, VH/CD and MLT compared with the other groups (p < 0.05). Dietary ME and CP levels greatly influenced cecal microbiota composition. Chickens in the MEMP group exhibited an increased abundance of Erysipelotrichaceae, Syntrophomonadaceae, Akkermansia, and Clostridia_vadinBB60_group, and there was an obvious decrease in the relative abundance of Desulfobacterota (p < 0.05). This study demonstrated that dietary ME and CP levels, along with their interactions, could significantly influence the growth performance, apparent nutrient metabolism, and intestinal development of Jingyuan chickens during the growing phase. Dietary ME and CP levels modulated the cecal microbiota composition, potentially inhibiting the abundance of harmful bacteria Desulfobacterota, while enriching the abundance of beneficial bacteria, thereby enhancing gut development and nutrient absorption. The combination of medium-level ME and CP (11.70 MJ/kg ME, 15.50% CP) demonstrated the most favorable outcomes in our study. Full article

Given the restrictions on animal growth promoters, alternative plant-based additives—particularly those rich in phenolic compounds, such as agro-industrial by-products—have been explored. These additives help to mitigate heat stress, which negatively affects productivity by impacting intestinal health and antioxidant status. This study evaluated the effects of individual and combined supplementation of ferulic acid (FA) and grape pomace (GP) on antioxidant enzyme activity, as well as intestinal histomorphometry, in finishing pigs under heat stress. Forty Yorkshire × Duroc pigs were randomly assigned to four treatments: control, 25 mg/kg FA, 2.5% GP, and MIX (FA + GP). FA supplementation increased intestinal villus height, while GP increased villus width in the duodenum and jejunum (p < 0.05). Antioxidant enzyme activity (SOD, CAT, and GPx) increased in pigs supplemented with GP (p < 0.05). These results suggest that GP and FA have potential as functional additives in monogastric diets, improving intestinal health and muscle antioxidant status and contributing to growth modulation. Full article

Sheep have been widely used as a model for osteoporosis research. This study aimed to characterise changes in microstructure and composition in lumbar vertebrae L1–L7 and the proximal femur after implementation of a bone loss induction protocol (in this species). A sham control and experimental group (glucocorticoid-treated ovariectomized sheep) were used (n = 6/group), with a study duration up to the 24th postoperative week. Through micro-computed tomography, vertebrae and femoral head trabecular bones from the experimental group presented a consistent decrease in bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N) and an increase in trabecular separation (Tb.Sp) and total porosity (p > 0.05). The mineral density of the femoral heads from the experimental group showed a statistically significant decrease (p ˂ 0.05). The entire histomorphometric analysis of the vertebrae in the experimental group showed an increase in cortical porosity (Ct.Po) and a decrease in cortical thickness (Ct.Th) (p ˂ 0.0001 and p ˂ 0.001, respectively). Vertebrae L6 and L7 were the most affected, showing a significant increase in Ct.Po (p < 0.05) and a significant decrease in Ct.Th at the L6 level (p < 0.05). Regarding the trabecular bone at the vertebral level, only L4 showed a significant increase in Tb.Sp (p ˂ 0.05). In the femoral heads’ subchondral cortical layer, the Ct.Po increased significantly and Ct.Th decreased (p < 0.01), and at the trabecular level, the BV/TV, Tb.Th, and Tb.N decreased significantly, while Tb.Sp increased (p < 0.05). In conclusion, the L4, L6 and L7 vertebrae seem the most suitable for further preclinical and translational studies of vertebral augmentation or spinal fusion in this animal model. Full article

Osteosarcopenia, characterized by concurrent bone loss and muscle wasting, significantly impacts mobility and quality of life. While age-related primary osteosarcopenia is well-studied, secondary osteosarcopenia (SOS) caused by chronic diseases remains poorly understood, particularly in young individuals. The present study aimed to comprehensively characterize musculoskeletal alterations associated with SOS using a juvenile porcine model of cerulein-induced chronic pancreatitis. Femoral bone analysis included densitometry, mechanical testing, histomorphometry, and serum bone turnover markers. The quadriceps femoris muscle was evaluated through histological analysis and gene expression profiling of antioxidant enzymes and apoptotic regulators. Animals with SOS showed significantly reduced femoral BMD compared to controls, with altered cortical geometry and compromised mechanical properties. Trabecular bone analysis revealed classic osteoporotic changes with decreased bone volume fraction. Negative changes were also observed in the growth plate morphology, indicating impaired endochondral ossification. Bone turnover markers indicated elevated bone resorption and altered formation. Muscle analysis demonstrated sarcopenic changes with selective atrophy of fast-twitch type II fibers and increased fiber density. At the molecular level, SOS muscles exhibited downregulated expression of CAT and CASP3, suggesting muscle atrophy predominantly mediated by oxidative stress and caspase-independent proteolysis rather than classical apoptosis. In conclusion, chronic pancreatitis in young pigs induces coupled bone and muscle degeneration consistent with secondary osteosarcopenia, demonstrating that muscle–bone crosstalk dysfunction occurs early in chronic inflammatory disease. Full article

Platelet-rich plasma (PRP) has become an increasingly valuable biologic approach for personalized regenerative medicine because of its potent anti-inflammatory/healing effects. It is thought to be an excellent source of growth factors that can promote tissue healing and lessen fibrosis. Although this treatment has demonstrated effectiveness in numerous disease areas, its impact on pulmonary fibrosis (PF) caused by silver nanoparticles (AgNPs) via its antiapoptotic effects remains to be explored. AgNPs were synthesized biologically by Bacillus megaterium ATCC 55000. AgNP characterization was carried out via UV–Vis spectroscopy, X-ray diffraction (XRD), dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) imaging to reveal monodispersed spheres with a mean diameter of 45.17 nm. A total of 48 male Wistar rats divided into six groups, with 8 rats per group, were used in the current study on the basis of sample size and power. The groups used were the PRP donor, control, AgNP, AgNP + PRP, AgNP + dexamethasone (Dexa) rat groups, and a recovery group. Body weights, hydroxyproline (HP) levels, and CASP3 and TWIST1 gene expression levels were assessed. H&E and Sirius Red staining were performed. Immunohistochemical studies for inducible nitric oxide synthase (iNOS) and cluster of differentiation 68 (CD68) with histomorphometry were conducted. A significant reduction in body weight (BWt) was noted in the AgNP group compared with the AgNP + PRP group (p < 0.001). HP, CASP3, and TWIST1 expression levels were significantly increased by AgNPs but decreased upon PRP (p < 0.001) treatment. Compared with those in the control group, the adverse effects of AgNPs included PF, lung alveolar collapse, thickening of the interalveolar septa, widespread lymphocytic infiltration, increased alveolar macrophage CD68 expression, and iNOS positivity in the cells lining the alveoli. This work revealed that PRP treatment markedly improved the histopathological and immunohistochemical findings observed in the AgNP group in a manner comparable to that of the Dexa. In conclusion, these results demonstrated the therapeutic potential of PRP in a PF rat model induced via AgNPs. This study revealed that PRP treatment significantly improved the histopathological and immunohistochemical alterations observed in the AgNP-induced group, with effects comparable to those of the Dexa. In conclusion, these findings highlight the therapeutic potential of PRP in a rat model of AgNP-induced PF. Full article

Background: Numerous experimental studies aim to improve outcomes of peripheral nerve repair following trauma. This study evaluates the efficacy of the human epineural patch (hEP) compared to the human amniotic membrane (hAM) in promoting nerve regeneration following sciatic nerve crush injury. Methods: Thirty-six athymic nude rats were divided into three groups (n = 12 per group) following nerve crush: (1) an unprotected injury site; (2) crush injury wrapped with hEP; and (3) crush injury wrapped with hAM. Animals were assessed over 6 or 12 weeks post-injury. Evaluations included motor recovery (Toe-Spread test), sensory recovery (Pinprick test), muscle denervation atrophy (the gastrocnemius muscle index (GMI)), histomorphometry (myelin thickness, axonal density, fiber diameter, and percentage of myelinated fibers), and immunofluorescence (GFAP, Laminin B, NGF, S-100, VEGF, vWF, HLA-DR, and HLA-I) assessments. Results: The hEP group showed superior motor recovery, axonal density and higher GMI values compared to the hAM and control groups. The increased expression of neurogenic and angiogenic markers highlighted its neuroregenerative potential. Negligible HLA-DR and HLA-I expression confirmed the lack of hEP and hAM immunogenicity. Conclusions: The application of hEP following sciatic nerve crush injury facilitated nerve regeneration, improved functional outcomes, and offered a viable alternative to hAM. Structural stability and the regenerative capacity position hEP as a new, promising off-the-shelf product for nerve regeneration. Full article

Obesity is considered a metabolic disease, in which leptin is used as an indicator of energy in the body. This hormone, in turn, is related to the neuroendocrine regulation of the reproductive axis. However, leptin excess secretion due to obesity can have a negative effect on reproduction. Overweight and obesity were induced through high-calorie diets. Lee and gonadosomatic indices were determined to characterize the model and degree of reproductive development in the testis and epididymis. Sperm quality was analyzed using spermatobioscopy. Morphometry was analyzed through histological analysis. The changes described affect testicular function in hormone and sperm production. Exposure of 3-month-old male rats to diets with different fat contents (10% and 60%) induced both overweight and obesity. The animals showed morphological alterations, both testicular and epididymal, the latter being more sensitive to dietary changes by modifying the epididymal index, morphometric parameters (in both organs), and a decrease in cilia length. These changes induced a reduction in sperm viability, as well as an increase in malformed spermatozoa. In conclusion, both overweight and obesity have effects on male reproduction by modifying the morphology and physiology of reproductive organs. Full article