Biointerface-engineered trabecular scaffolds integrating chitosan/icariin self-assembly and mild photothermal regulation for enhanced osteogenesis.

High-yield cell-derived extracellular matrix bioink via macromolecular crowding for versatile 3D bioprinting.

Osteoporosis is a major health risk for postmenopausal women and is primarily driven by estrogen decline-induced bone loss. As critical post-transcriptional regulators of gene expression, dysregulated microRNAs (miRs) participate in the pathogenesis of osteoporosis by disrupting key signaling pathways that govern bone homeostasis and remodeling. This study investigated the functional role of the miR-33b-5p/ABCA1 axis in cellular and murine models of osteoporosis. Alizarin Red S staining and alkaline phosphatase (ALP) activity assays were performed to detect osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). ABCA1 mRNA and protein levels, as well as osteogenic proteins such as RUNX2, OPN, and OCN, were quantified by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. Dexamethasone (DEX)-stimulated BMSCs were employed as an in vitro osteoporosis model. To overexpress ABCA1 in BMSCs, the cells were transfected with pcDNA3.1 vectors carrying the full-length ABCA1 coding sequence. ABCA1 silencing was accomplished using short hairpin RNA targeting ABCA1 (sh-ABCA1), whereas miR-33b-5p inhibition was achieved using a specific miR-33b-5p inhibitor. The direct interaction between miR-33b-5p and its downstream target ABCA1 was validated using luciferase reporter assays. After an ovariectomized (OVX) mouse model was established, micro-CT was performed to detect bone density by assessing trabecular thickness and trabecular number in femurs. ABCA1 expression increased during osteogenic differentiation of BMSCs, suggesting an essential role in this process. In DEX-treated BMSCs, the ABCA1 level was downregulated. ABCA1 overexpression promoted osteoblast differentiation, as evidenced by increased mineralized nodule formation, upregulation of key osteogenic proteins, and elevated ALP activity. MiR-33b-5p directly interacted with the 3'UTR of ABCA1, with the binding region conserved across multiple species. MiR-33b-5p inhibitors promoted osteogenic differentiation, and this effect was abolished upon ABCA1 depletion via sh-ABCA1. Animal experiments indicated that miR-33b-5p inhibition promoted trabecular bone formation in the femurs of OVX mice, including increased trabecular number and thickness, along with elevated expression of osteoblastic markers. A significant negative correlation between miR-33b-5p and ABCA1 expression levels was observed in bone tissues of experimental mice. . Inhibition of miR-33b-5p promotes osteoblastic differentiation by regulating ABCA1 in experimental osteoporosis models.

Pulp necrosis in immature permanent teeth arrests root development and compromises long-term prognosis. This study aimed to develop a multifunctional scaffold integrating structural biomimicry, mechanical matching, and sustained growth factor release for orderly root regeneration. A poly(ε-caprolactone) (PCL) conical scaffold was fabricated via melt electrowriting (MEW) combined with mechanical winding. Bone morphogenetic protein‑2 (BMP‑2)-loaded microspheres were prepared and physically incorporated into the scaffold. The scaffold surface was modified with collagen. Human dental pulp stem cells (hDPSCs) were cultured on the scaffold to evaluate proliferation, adhesion, and osteogenic differentiation. The scaffold exhibited a trilayer "collagen-microsphere-PCL" architecture with mechanical compatibility (elastic modulus: 22.5MPa; fracture strength: 5.29MPa; elongation: 441.59%). Microspheres (2.86 ± 0.45μm) showed a gradient distribution and sustained release (70-75% over 90 days). In vitro, the scaffold promoted hDPSC adhesion and proliferation and significantly enhanced osteogenic differentiation with elevated alkaline phosphatase activity, upregulated the expression of osteogenic-related genes, and increased protein levels. The scaffold integrates structural support, controlled growth factor delivery, and a bioactive interface, offering a promising strategy for root development in immature permanent teeth. By enabling physiological root development, the scaffold addresses a critical unmet need, offering a viable alternative to conventional root canal therapy.

This study investigates anodic surface modification of the Ti 47 Cu 38 Zr 7.5 Fe 2.5 Sn 2 Si 1 Ag 2 bulk metallic glass in a nontoxic potassium pyrophosphate electrolyte and its impact on corrosion behavior and cytocompatibility. The treatment forms bilayered oxide films (20–90 nm) via selective Cu dealloying and Ti/Zr oxide formation. Analyses by transmission electron microscopy and glow discharge optical emission spectroscopy reveal a dense inner amorphous layer and a porous outer layer containing metallic Cu nanocrystals, formed through Cu mobilization and reduction. Electrochemical testing demonstrates that anodization at 1.3 V vs. saturated mercury sulfate electrode significantly enhances corrosion resistance in chloride‐containing phosphate‐buffered saline and reduces Cu ion release by approximately 50% compared to the untreated state. Contact angle measurements confirm increased surface hydrophilicity due to the Ti/Zr oxide matrix. Biological evaluation shows that this optimized surface promotes human bone marrow stromal cell spreading and focal adhesion formation. It results in a 4.5‐fold increase in cell proliferation and elevated activity of the osteogenic marker tissue nonspecific alkaline phosphatase, indicating enhanced osteogenic differentiation. These findings highlight that controlled anodization in a nontoxic pyrophosphate electrolyte can tailor surface oxide structure and composition, simultaneously improving corrosion resistance, cytocompatibility of Ti–Cu‐based metallic glasses for advanced biomedical implants.

Systemic inflammation impairs bone health by inhibiting the osteogenic differentiation of bone marrow stromal cells (BMSCs), thereby contributing to bone loss. Diallyl disulfide (DADS), a natural compound with anti-inflammatory properties, was investigated for its ability to mitigate inflammatory bone loss (IBL), reverse LPS-induced suppression of osteogenic differentiation, and elucidate the underlying mechanisms. An LPS-induced mouse model of IBL was used to evaluate the effects of DADS by micro-CT, histological and immunohistochemical staining, and serum ELISA. An in vitro model was established by exposing BMSCs to LPS. The optimal concentration of DADS was determined using a Cell Counting Kit-8 (CCK-8) assay. Osteogenic differentiation was evaluated by alkaline phosphatase (ALP) staining, Alizarin Red S (ARS) staining, immunofluorescence, and Western blot. Network pharmacology and transcriptome sequencing were employed to identify potential therapeutic mechanisms, and the PI3K/AKT pathway was verified using the inhibitor LY294002. Micro-CT and histological analyses confirmed that DADS attenuated bone loss in the IBL mouse model. Toluidine blue staining and immunohistochemical analysis demonstrated that DADS promoted osteogenic differentiation. Immunohistochemical detection and serum ELISA of inflammatory cytokines revealed that DADS significantly reduced inflammatory levels in IBL mice. In vitro, the LPS-induced inhibition of osteogenic differentiation in BMSCs was reversed by DADS. Following DADS treatment, elevated ALP activity, matrix mineralization, and osteogenic marker expression, and reducing inflammatory mediators were observed. Network pharmacology and transcriptome sequencing revealed that DADS may exert its effects through the PI3K/AKT signaling pathway. Subsequent experiments conclusively established that DADS activated the PI3K/AKT signaling pathway. The osteogenic differentiation induced by DADS was inhibited by the PI3K inhibitor LY294002, indicating that the PI3K/AKT pathway is crucial in the osteogenic differentiation promoted by DADS. DADS counteracts LPS-induced bone loss by promoting osteogenic differentiation via PI3K/AKT activation, highlighting its therapeutic potential for inflammatory bone diseases.

Serpinh1 promotes fracture healing by enhancing osteogenesis via activation of the Wnt/β-catenin signaling pathway.

Supercritical CO2-foamed hierarchically porous PLA/PBS-based scaffold for advanced bone regeneration.

The combination of structural-functional hepatic immaturity, increased permeability of biological barriers, and concomitant pathology in premature newborns with clinical signs of neonatal jaundice (NJ) gives rise to distinctive patterns of metabolic dysregulation, necessitating systematic monitoring of blood serum enzyme activity. Comprehensive evaluation of hepatobiliary system (HS) functional status indicators — in particular, enzyme activity markers — enables optimization of diagnostic and therapeutic strategies through clarification of the pathophysiological mechanisms underlying NJ development, thereby preventing irreversible metabolic and neurological sequelae. The aim: To perform a comprehensive analysis of blood serum enzyme activity as a marker of hepatobiliary system (HS) functional status in premature newborns presenting with clinical signs of neonatal jaundice (NJ). Materials and methods: A total of 85 premature newborns of 34–36 weeks of gestational age presenting with clinical signs of NJ in the context of perinatal pathology were examined. The newborns were assessed and allocated into two subgroups based on condition severity: group A comprised 28 newborns with perinatal pathology of moderate severity, and group B comprised 30 newborns with severe perinatal pathology. The comparison group comprised 27 newborns of appropriate gestational age with a satisfactory course of postnatal adaptation. The following blood serum indicators were assessed: total bilirubin (TB) and bilirubin fractions, as well as the activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH). The study was approved by the Biomedical Ethics Commission of Bukovinian State Medical University (BSMU) (Protocol No. 2, dated February 9, 2015). Prior to study initiation, written informed consent was obtained from the parents, with explanation of the purpose, objectives, and methods of the laboratory investigation. Statistical analysis was performed using STATISTICA (StatSoft Inc., USA; Version 10.0). Quantitative comparisons against reference values were conducted using Student's t-test (p < 0.05). Receiver operating characteristic (ROC) curve analysis was performed using MedCalc Statistical Software (Statistical Software Package for Biomedical Research; Version 16.1; 2023). The dissertation work was conducted as part of the research activity of the Department of Pediatrics, Neonatology, and Perinatal Medicine of BSMU on the topics "Improvement of Prognostication, Diagnostics and Treatment of Perinatal Pathology in Newborns and Early-Age Children, Optimization of Catamnestic Observation and Rehabilitation Schemes" (state registration No. 0115U002768) and "Chronobiological and Adaptation Aspects and Features of Vegetative Regulation of Pathological Conditions in Children of Different Ages" (state registration No. 0122U002245). Results and discussion. The most informative markers for confirming enzyme activity dysregulation in the context of HS dysfunction with concurrent clinical signs of NJ in perinatal pathology of moderate severity were elevated GGT activity exceeding 116.65 U/L and LDH activity exceeding 776.41 U/L. These threshold values, established by ROC analysis, demonstrated the highest sensitivity (SN) and specificity (SP) of 100.0%. Elevated ALP activity exceeding 376 U/L also demonstrated high diagnostic value (SN 93.33%, SP 100.0%). ALT and AST activity indicators (SN 63.33% and 53.33%, respectively) did not achieve sufficient discriminatory capacity in newborns with moderate-severity perinatal pathology presenting with NJ; however, both markers demonstrated maximal specificity for exclusion of the pathological process (SP 100.0%). In late preterm newborns with persistent NJ signs and severe perinatal pathology, LDH activity emerged as the defining diagnostic parameter. The conventionally accepted indicators — specifically ALT, AST, GGT, and ALP activities — did not demonstrate sufficient associative significance or prognostic value in NJ associated with severe perinatal pathology. To enhance the efficacy of clinical and diagnostic measures in NJ during the acute phase of perinatal pathology in late preterm newborns, further investigation of additional laboratory criteria characterizing enzyme activity — as a critical determinant of metabolic dysregulation arising from HS dysfunction — is warranted. Conclusions. 1. Assessment of enzyme activity markers — specifically ALT, AST, GGT, ALP, and LDH — constitutes an essential component of the recommended laboratory evaluation of blood serum in newborns presenting with clinical signs of NJ in the context of perinatal pathology. 2. The laboratory criteria of enzyme activity dysregulation in severe-condition premature newborns are elevated GGT activity exceeding 116.65 U/L and LDH activity exceeding 776.41 U/L, which confirm the presence of HS dysfunction. 3. The conventionally accepted indicators — specifically ALT, AST, GGT, and ALP activities — did not demonstrate sufficient associative significance or prognostic value in NJ associated with severe perinatal pathology by ROC analysis, thereby justifying the need for further scientific investigation aimed at identifying additional diagnostic criteria.

IntroductionPolyetheretherketone (PEEK) is a promising orthopedic implant material due to its bone-mimetic mechanical properties; however, its bioinert surface and susceptibility to bacterial colonization limit its clinical efficacy. Strategies to enhance osteointegration while providing antibacterial functionality are urgently needed.MethodsGraphene oxide (GO) was functionalized with polyethylene glycol (PEG) and complexed with a plasmid encoding bone morphogenetic protein-2 (pBMP-2) to form a GO-PEG/pBMP-2 complex. This complex was coated onto sulfonated PEEK (SPEEK) via freeze-drying, yielding SPEEK-(GO-PEG/pBMP-2) composites. The materials were systematically compared with pristine PEEK, SPEEK, and SPEEK-GO-PEG controls. Surface chemistry, morphology, hydrophilicity, gene loading efficiency, and coating uniformity were characterized. In vitro assessments included cell adhesion, proliferation, viability, alkaline phosphatase (ALP) activity, mineralized matrix deposition, osteogenic gene expression, and antibacterial activity against Staphylococcus aureus and Escherichia coli.ResultsSuccessful PEGylation of GO and efficient pBMP-2 loading were confirmed, with uniform coating and significantly improved hydrophilicity on SPEEK-(GO-PEG/pBMP-2). This composite markedly enhanced osteoblast adhesion, proliferation, and viability, along with elevated ALP activity, increased mineralized nodule formation, and upregulated expression of key osteogenic genes (e.g., Runx2, OPN, OCN). Both GO-containing coatings—SPEEK-GO-PEG and SPEEK-(GO-PEG/pBMP-2)—exhibited strong antibacterial effects against both Gram-positive and Gram-negative bacteria, with no significant difference attributable to BMP-2 loading.ConclusionThe SPEEK-(GO-PEG/pBMP-2) platform simultaneously promotes osteogenesis through localized BMP-2 gene delivery and provides robust antibacterial protection via GO. This dual-functional design addresses two major limitations of PEEK implants, offering a promising strategy for next-generation orthopedic biomaterials.

Serum biochemical parameters are reliable indicators of sub-lethal toxicity and organ dysfunction in fish exposed to environmental contaminants. This study evaluated the serum biochemical responses of Oreochromis niloticus juveniles exposed to aqueous extracts of Leptadania hastata, Nymphaea lotus, and urea fertilizer under a 96-hour static bioassay. Fish were subjected to graded concentrations of each test substance, while a control group was maintained without exposure. Serum total protein, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were analyzed using standard diagnostic procedures. Results showed significant (p<0.05) concentration-dependent alterations in all biochemical parameters. The control group consistently recorded higher total protein and albumin levels and more stable enzyme activities compared to exposed fish. Leptadania hastata induced the most pronounced biochemical disruptions, followed by urea fertilizer, while Nymphaea lotus exerted comparatively milder effects. Elevated ALP activity in urea fertilizer treatments suggests hepatobiliary stress associated with nitrogenous toxicity, whereas reductions in protein parameters across treatments indicate impaired hepatic synthetic function. These findings demonstrate that short-term exposure to plant extracts and urea fertilizer can compromise liver function and metabolic stability in Oreochromis niloticus juveniles. It is therefore recommended that the use of medicinal plant extracts and urea fertilizer in aquaculture systems be carefully regulated, with routine biochemical monitoring to prevent sub-lethal physiological damage in cultured fish.

Accelerating bone healing in femoral defect model using FBXO6-modified bone marrow-derived mesenchymal stem cells on a collagen scaffold.

Acute liver injury (ALI) remains a critical health challenge driven by oxidative stress, inflammation, and apoptosis. This study aimed to evaluate whether astaxanthin (ASX) and/or selenium (Se) can prevent carbon tetrachloride (CCl₄)-induced ALI in rats and to explore the transcriptional regulation of antioxidant, inflammatory, and apoptotic pathways, with particular focus on nuclear factor erythroid 2-related factor 2 (Nrf2), reactive oxygen species (ROS), and nuclear factor kappa-light-chain-enhancer of activated B cells (Nfkb1). Nrf2/ROS/Nfkb1 and Bcl-2-associated X protein (Bax) and B-cell lymphoma 2 (Bcl-2) signaling pathways. Sixty male Sprague-Dawley rats were randomly assigned to ten groups (n = 6): control, SIL (silymarin), ASX (astaxanthin), Se (selenium), ASX + Se (astaxanthin + selenium), CCl4 (carbon tetrachloride), and their corresponding pretreatment counterparts. ALI was induced with a single intraperitoneal CCl4 injection (1mL/kg). Serum biochemical markers of hepatic function, oxidative and inflammatory mediators, and apoptosis-related genes were quantified. Molecular expression of Nrf2, Nfkb1, Bax, Bcl-2, caspase-3, and related cytokines was assessed. Liver histology was examined to confirm tissue-level alterations. CCl4 administration significantly elevated serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) activities, while suppressing antioxidant and cytoprotective mediators, nuclear factor erythroid 2-related factor 2 (Nrf2), superoxide dismutase (sod), and interleukin-10 (IL-10), and upregulating pro-inflammatory and pro-apoptotic markers (nuclear factor kappa-light-chain-enhancer of activated B cells (Nfkb1), cyclooxygenase-2 (COX-2), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), inducible nitric oxide synthase (iNOS), Bcl-2-associated X protein (Bax), and caspase-3). Histopathology showed necrosis, inflammation, and steatosis. Pretreatment with ASX and/or Se markedly restored biochemical parameters, modulated gene expression toward antioxidant and anti-inflammatory profiles, and significantly improved hepatic architecture, with the combination therapy exerting the most pronounced effects. ASX and Se effectively mitigate CCl4-induced ALI by enhancing antioxidant defense, suppressing inflammation, and inhibiting apoptosis through modulation of key signaling pathways. These findings support their potential as complementary agents for hepatoprotection.

Osteoarthritis is a degenerative joint disorder characterized by progressive cartilage breakdown and altered bone metabolism. This study aimed to evaluate the correlation between serum alkaline phosphatase (ALP) activity and serum calcium levels in patients with osteoarthritis and to compare these parameters with those of healthy controls to identify deviations from normal biochemical values. A total of 100 osteoarthritis patients (61% male and 39% female), aged 45 years and above, attending the outpatient department of Teaching Hospital Shahdara were included in the study. Serum samples were analyzed for ALP activity and calcium concentration using standard biochemical methods. The results demonstrated significantly elevated serum ALP activity in osteoarthritis patients, while serum calcium levels were comparatively reduced. An inverse correlation was observed between serum ALP activity and serum calcium levels. These findings suggest that alterations in ALP and calcium metabolism are associated with osteoarthritis progression. Measurement of serum ALP and calcium may serve as simple, cost-effective biochemical biomarkers for assessing disease severity in patients with osteoarthritis.

The design of biomaterial scaffolds for bone tissue engineering requires a balance between bioactivity, porosity, mechanical stability, and osteoinductivity. Kappa- (KC) and iota-carrageenan (IC) have been explored for scaffold fabrication due to their biocompatibility and structural similarity to glycosaminoglycans. However, there are limited reports on how their distinct sulfation degree affects the osteogenic differentiation of cells cultured on them. While laponite has been reported as an osteoinductive nanoclay, its combined effect with different carrageenan types and its concentration-dependent effect on scaffold functionality remain unexplored. Therefore, we developed composite scaffolds comprising poly(vinyl alcohol) (PVA) and gelatin (GEL), reinforced with kappa- or iota-carrageenan (KC, IC) and functionalized with two different concentrations of laponite (LAP), 0.5 and 1% w/v, to monitor composition-structure-function relationships. The scaffolds were fabricated via lyophilization and dual crosslinking, and characterized for their physicochemical, structural, mechanical, and biological properties. The incorporation of both carrageenans into scaffolds, maintained high swelling ratios of 600% after 24 h, and increased porosity without altering their apparent density (0.09-0.11 g/cm3), whereas LAP preserved interconnectivity, densified pore walls, raised their compressive modulus at >220 kPa, and improved stability (>60% mass retained after 40 days). In vitro validation using MC3T3-E1 pre-osteoblastic cells demonstrated robust cytocompatibility, with the LAP-containing scaffolds significantly promoting cell adhesion, proliferation, and osteogenic differentiation, evidenced by elevated alkaline phosphatase activity, calcium production and collagen secretion. Direct comparison between KC and IC scaffolds confirmed that differences in sulfate substitution modulated scaffold stiffness, swelling, and degradation, while variation in LAP concentration affected the biological response, with the 0.5 wt% concentration favoring early cell proliferation, whereas the 1 wt% significantly promoted the osteogenic differentiation. This compositional strategy demonstrates how tuning the interplay between carrageenan and laponite can balance scaffold hydration, mechanical and biological properties, thereby guiding the design of scaffolds for bone repair.

ZIF-67-anchored halloysite nanotubes infused in chitosan hydrogel for enhanced bone tissue engineering.

Oral and maxillofacial surgeries or trauma (e.g. impacted tooth extraction, jaw fracture or tumor resection) often lead to concurrent peripheral nerve injury and bone defects, while current collagen/gelatin sponges offer limited therapeutic effects. To address this challenge, we developed innovative electrospun scaffolds (MOF2, MOF4 and MOF6) by in situ synthesis of 3,5-dihydroxybenzoic acid/zinc (DHBA/Zn-MOF) hybrids within a gelatin/polycaprolactone matrix. In vitro, Schwann cells treated with material extracts exhibited enhanced migration, regulated myelin-associated genes (Ngf/Pmp22 upregulated, Ncam downregulated) and increased NGF protein expression via the PI3K pathway. Co-cultured PC12 cells showed increased neurite outgrowth, confirming neural repair potential. Osteoblasts exposed to material extracts showed elevated alkaline phosphatase activity, enhanced mineralization and upregulated osteogenic genes (Runx2, Alp and Opg), verifying osteogenic capacity. In vivo, MOF6 scaffolds achieved superior motor function recovery in a rat sciatic nerve crush model (evidenced by increased compound muscle action potentials and reduced gastrocnemius muscle atrophy) and promoted trabecular bone formation in a rat skull defect model (validated by micro-CT and histological analyses). These findings underscore the dual-functional capability of DHBA/Zn-hybrid scaffolds to simultaneously promote nerve repair and bone regeneration, offering a promising therapeutic approach for complex neuro-bone composite injuries in clinical practice.

This study investigated the effects of a polycaprolactone (PCL)-polyethylene glycol (PEG) scaffold incorporated with concentrated growth factor (CGF) on the adhesion, proliferation, and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The PCL-PEG-CGF composite scaffold was fabricated using an immersion and freeze-drying technique. Its microstructure, mechanical properties, and biocompatibility were systematically characterized. The hPDLSCs were isolated through enzymatic digestion, and the hPDLSCs were identified through flow cytometry. Third-passage hPDLSCs were seeded onto the composite scaffolds, and their adhesion, proliferation and osteogenic differentiation were assessed using CCK-8 assays, 4',6-diamidino-2-phenylindole (DAPI) staining, alkaline phosphatase (ALP) staining, alizarin red staining, and Western blot analysis of osteogenesis-related proteins [Runt-related transcription factor 2 (Runx2), ALP, and morphogenetic protein 2 (BMP2)]. Scanning electron microscopy revealed that the PCL-PEG-CGF composite scaffold exhibited a honeycomb-like structure with heterogeneous pore sizes. The composite scaffold exhibited excellent hydrophilicity, as evidenced by a contact angle (θ) approaching 0° within 6 s. Its elastic modulus was measured at (4.590 0±0.149 3) MPa, with comparable hydrophilicity, fracture tensile strength, and fracture elongation to PCL-PEG scaffold. The hPDLSCs exhibited significantly improved adhesion to the PCL-PEG-CGF composite scaffold compared with the PCL-PEG scaffold (P<0.01). Additionally, cell proliferation was markedly improved in all the experimental groups on days 3, 5, and 7 (P<0.01), and statistically significant differences were found between the PCL-PEG-CGF group and other groups (P<0.01). The PCL-PEG-CGF group showed significantly elevated ALP activity (P<0.05), increased mineralization nodule formation, and upregulated expression of osteogenic-related proteins (Runx2, BMP2 and ALP; P<0.05). The PCL-PEG-CGF composite scaffold exhibited excellent mechanical properties and biocompatibility, enhancing the adhesion and proliferation of hPDLSCs and promoting their osteogenic differentiation by upregulating osteogenic-related proteins.

Gastroenteritis is one of the most common gastrointestinal disorders in dogs, recorded among representatives of all breeds and age groups, with such clinical signs as anorexia, depression, vomiting, and diarrhea. These symptoms are complex in nature, encompassing both local gastrointestinal disturbances and systemic changes caused by dehydration and intoxication. In this study, for the first time, the diagnostic informativeness of a methodological approach based on determining the rehabilitation potential of sick animals was analyzed. The criteria for this assessment include hematological and biochemical blood parameters. Rehabilitation potential is an indicator of the animal's potential ability to achieve complete recovery within a certain period of time. This method was first proposed and implemented in 2024–2025 at the Department of Internal Diseases and Clinical Diagnostics of Animals of the State Biotechnological University during the examination of clinically healthy dogs kept in an animal shelter. Determining the rehabilitation potential made it possible to qualitatively and quantitatively assess the adaptive capabilities of dogs’ organisms during long-term shelter maintenance. Previously, this methodological approach had not been applied to the examination of animals suffering from gastroenteritis. Analysis of laboratory test results revealed a significant decrease in hemoglobin, erythrocytes, and eosinophilic leukocytes, as well as an increase in the total number of leukocytes. Biochemical studies showed elevated activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), along with reduced levels of total protein, albumin, and potassium in the blood of affected dogs. These changes were caused by inflammatory processes of the gastric and intestinal mucosa, dehydration, hepatic hypoxia, and the action of toxic substances due to intestinal barrier dysfunction. Calculations of the degree of rehabilitation potential provided an integral assessment of the health status of sick dogs and the potential adaptive capacity of their organisms, which was determined to be low. The obtained results may be useful for developing effective treatment methods for dogs with gastroenteritis. The informativeness of the animals’ rehabilitation potential, based on hematological and biochemical serum parameters, is emphasized.

This study investigated the delivery of small interfering RNA (siRNA) silencing prolyl hydroxylase domain 2 (PHD2) via poly(lactic-co-glycolic acid) (PLGA)-bioactive glass scaffold to improve angiogenesis and osteogenesis for biomineralized tissue engineering. Polyethylenimine (PEI) was complexed with PHD2siRNA at 3:1, 6:1, and 8:1 ratios, and the resulting complexes were characterized by transmission electron microscopy, NanoSight analysis, and gel retardation assays. Their biological effects on human umbilical vein endothelial cells (HUVECs) were evaluated to determine the optimal ratio. PLGA-bioactive glass scaffolds were prepared and loaded with the most effective PEI/PHD2siRNA complex. HUVECs were used to confirm controlled RNA release from the scaffolds. The angiogenic effects of HUVECs and osteogenic differentiation of stem cells derived from human exfoliated deciduous teeth (SHED) were evaluated. The 8:1 ratio of PEI/PHD2siRNA demonstrated the highest efficiency, improving HUVEC tube formation. Incorporating these complexes into PLGA-bioactive glass scaffolds confirmed RNA release, as indicated by improved tube formation. The scaffolds promoted the proliferation and angiogenic differentiation of HUVECs. SHED cells cultured on the scaffolds demonstrated improved proliferation and osteogenic differentiation, evidenced by elevated alkaline phosphatase activity and mineral deposition. Coculturing SHED cells with HUVECs on the scaffolds exerted synergistic effects, producing more robust osteogenic differentiation than culturing SHED cells alone. Immunofluorescence staining revealed increased expression of both angiogenic and osteogenic markers. PHD2 siRNA delivery via PLGA-bioactive glass scaffolds enhanced angiogenesis and osteogenesis, supporting its potential in tissue engineering. This dual-function scaffold offers a promising strategy for dental and craniofacial regeneration.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-24808-9.