Increased Alkaline Phosphatase Research Articles

Polycystic Ovary Syndrome Physiologic Pathways Implicated Through Clustering of Genetic Loci.

Polycystic ovary syndrome (PCOS) is a heterogeneous disorder, with disease loci identified from genome-wide association studies (GWAS) having largely unknown relationships to disease pathogenesis. This work aimed to group PCOS GWAS loci into genetic clusters associated with disease pathophysiology. Cluster analysis was performed for 60 PCOS-associated genetic variants and 49 traits using GWAS summary statistics. Cluster-specific PCOS partitioned polygenic scores (pPS) were generated and tested for association with clinical phenotypes in the Mass General Brigham Biobank (MGBB, N = 62 252). Associations with clinical outcomes (type 2 diabetes [T2D], coronary artery disease [CAD], and female reproductive traits) were assessed using both GWAS-based pPS (DIAMANTE, N = 898,130, CARDIOGRAM/UKBB, N = 547 261) and individual-level pPS in MGBB. Four PCOS genetic clusters were identified with top loci indicated as following: (i) cluster 1/obesity/insulin resistance (FTO); (ii) cluster 2/hormonal/menstrual cycle changes (FSHB); (iii) cluster 3/blood markers/inflammation (ATXN2/SH2B3); (iv) cluster 4/metabolic changes (MAF, SLC38A11). Cluster pPS were associated with distinct clinical traits: Cluster 1 with increased body mass index (P = 6.6 × 10-29); cluster 2 with increased age of menarche (P = 1.5 × 10-4); cluster 3 with multiple decreased blood markers, including mean platelet volume (P = 3.1 ×10-5); and cluster 4 with increased alkaline phosphatase (P = .007). PCOS genetic clusters GWAS-pPSs were also associated with disease outcomes: cluster 1 pPS with increased T2D (odds ratio [OR] 1.07; P = 7.3 × 10-50), with replication in MGBB all participants (OR 1.09, P = 2.7 × 10-7) and females only (OR 1.11, 4.8 × 10-5). Distinct genetic backgrounds in individuals with PCOS may underlie clinical heterogeneity and disease outcomes.

Development of miR-26a-activated scaffold to promote healing of critical-sized bone defects through angiogenic and osteogenic mechanisms

Very large bone defects significantly diminish the vascular, blood, and nutrient supply to the injured site, reducing the bone's ability to self-regenerate and complicating treatment. Delivering nanomedicines from biomaterial scaffolds that induce host cells to produce bone-healing proteins is emerging as an appealing solution for treating these challenging defects. In this context, microRNA-26a mimics (miR-26a) are particularly interesting as they target the two most relevant processes in bone regeneration-angiogenesis and osteogenesis. However, the main limitation of microRNAs is their poor stability and issues with cytosolic delivery. Thus, utilising a collagen-nanohydroxyapatite (coll-nHA) scaffold in combination with cell-penetrating peptide (RALA) nanoparticles, we aimed to develop an effective system to deliver miR-26a nanoparticles to regenerate bone defects in vivo. The microRNA-26a complexed RALA nanoparticles, which showed the highest transfection efficiency, were incorporated into collagen-nanohydroxyapatite scaffolds and in vitro assessment demonstrated the miR-26a-activated scaffolds effectively transfected human mesenchymal stem cells (hMSCs) resulting in enhanced production of vascular endothelial growth factor, increased alkaline phosphatase activity, and greater mineralisation. After implantation in critical-sized rat calvarial defects, micro CT and histomorphological analysis revealed that the miR-26a-activated scaffolds improved bone repair in vivo, producing new bone of superior quality, which was highly mineralised and vascularised compared to a miR-free scaffold. This innovative combination of osteogenic collagen-nanohydroxyapatite scaffolds with multifunctional microRNA-26a complexed nanoparticles provides an effective carrier delivering nanoparticles locally with high efficacy and minimal off-target effects and demonstrates the potential of targeting osteogenic-angiogenic coupling using scaffold-based nanomedicine delivery as a new "off-the-shelf" product capable of healing complex bone injuries.

Clinical features, diagnostics and treatment of FGF23 secreting tumors: series of 40 clinical cases

Tumor-induced osteomalacia is an acquired rare disease manifested by hypophosphatemic osteomalacia due to excessive secretion of fibroblast growth factor 23 (FGF23). FGF 23 is a non-classical hormone secreted by bone tissue (osteocytes) and regulates phosphorus metabolism.The aim of this work is to present clinical experience in the diagnosis, treatment and rehabilitation of patients with tumor-induced osteomalacia. 40 patients with clinically-confirmed tumor-induced osteomalacia were included in the study, 34 of whom had the tumor localized, 27 underwent surgical treatment and 21 achieved stable remission. The median age was 48 [41; 63] years, 43% were men, the time left from the the onset of the disease was 8 [4; 10] years. Biochemical findings were hypophosphatemia 0.47 [0.4; 0.53] mmol/l, a decrease in the tubular reabsorption phosphate 62 [52; 67]%, and an increase in alkaline phosphatase of 183 [112; 294] units/l. At the time of diagnosis, 100% had multiple pathological fractures, only 10% could move independently, and 77.5% classified the pain as unbearable (8-10 points according to the 10-point pain syndrome scale ). Among the methods used to detect tumors, the most sensitive were scintigraphy with tectrotide with SPECT/CT 71.4% (20/28) and MRI 90% (18/20). In 35% of cases, the tumor was localized in soft tissues and in 65% in bone tissue; The tumor was most often detected in the lower extremities, followed by the head in frequency of localization. 18 patients currently have no remission and they receive conservative treatment (phosphorus and alfacalcidol n=15 and burosumab n=3). In case of achieving remission (n=21), regression of clinical symptoms and restoration of bone and muscle mass was observed. Extensive excision of the tumor without prior biopsy resulted in the best percentage of remission - 87%. Tumor-induced osteomalacia is characterized by severe damage to bone and muscle tissue with the development of multiple fractures, muscle weakness and severe pain syndrome. In laboratory diagnostics, attention should be paid to hypophosphatemia, a decrease in the tubular reabsorption phosphate index and increased alkaline phosphatase. The use of functional diagnostic methods with a labeled somatostatin analogue to the subtype 2 receptor and MRI with contrast enhancement are the most accurate methods of topical diagnostics. In case of localization of the tumor, a wide excision without a preliminary biopsy is recommended.

CeO2 Nanoparticle Bioactive Materials Promote MG-63 Osteogenic Differentiation and Antioxidant Activity Through NRF2 Signaling.

The incidence of bone-related diseases is higher in the elderly population, which greatly affects the patients' quality of life. Throughout this research, we synthesized a biocomposite nanomaterial of CeO2. The unique structural characteristics of CeO2 nanoparticles (CeO2 NPs) were studied by means of XRD, TEM, and SEM. Nanoparticles of an osteosarcoma cell line (MG-63) were assayed for ALP enzyme levels, key proteins in osteoblasts, and stained with Alizarin Red S to assess the physical properties, bioactivity, and calcium deposition of the osteosarcoma cell line. Moreover, we used H2O2 to construct an oxidative stress model to evaluate the antioxidant activity of CeO2 NPs. Experimental data showed that the CeO2 NPs increased the antioxidant capacity of MG-63 cells and significantly increased alkaline phosphatase activity, calcium deposition, and bone growth as manifested by increased expression of bone differentiation proteins BMP2, OCN, OPN, and type I collagen. Interestingly, RNA interference and functional recovery experiments confirmed that CeO2 NPs enhanced the antioxidant activity of MG-63 cells related to NRF2 signaling. In conclusion, the material is expected to be a potential treatment for bone-related diseases.

Bioactive Self-Regulated Liquified Microcompartments to Bioengineer Bone-Like Microtissues.

Designing a microenvironment that drives autonomous stromal cell differentiation toward osteogenesis while recapitulating the complexity of bone tissue remains challenging. In the current study, bone-like microtissues are created using electrohydrodynamic atomization to form two distinct liquefied microcapsules (mCAPs): i) hydroxypyridinone (HOPO)-modified gelatin (GH mCAPs, 7.5% w/v), and ii) HOPO-modified gelatin and dopamine-modified gelatin (GH+GD mCAPs, 7.5%+1.5% w/v). The ability of HOPO to coordinate with iron ions at physiological pH allows the formation of a semipermeable micro-hydrogel shell. In turn, the dopamine affinity for calcium ions sets a bioactive milieu for bone-like microtissues. After 21 days post encapsulation, GH and GH+GD mCAPs potentiate autonomous osteogenic differentiation of mesenchymal stem cells accompanied by collagen type-I gene upregulation, increased alkaline phosphatase (ALP)expression, and formation of mineralized extracellular matrix. However, the GH+GD mCAPs show higher levels of osteogenic markers starting on day 14, translating into a more advanced and organized mineralized matrix. The GH+GD system also shows upregulation of the receptor activator of nuclear factor kappa-B ligand (RANK-L) gene, enabling the autonomous osteoclastic differentiation of monocytes. These catechol-based mCAPs offer a promising approach to designing multifunctional and autonomous bone-like microtissues to study in vitro bone-related processes at the cell-tissue interface, angiogenesis, and osteoclastogenesis.

Zuo Gui Wan Promotes Osteogenesis via PI3K/AKT Signaling Pathway: Network Pharmacology Analysis and Experimental Validation.

Osteogenesis is vitally important for bone defect repair, and Zuo Gui Wan (ZGW) is a classic prescription in traditional Chinese medicine (TCM) for strengthening bones. However, the specific mechanism by which ZGW regulates osteogenesis is still unclear. The current study is based on a network pharmacology analysis to explore the potential mechanism of ZGW in promoting osteogenesis. A network pharmacology analysis followed by experimental validation was applied to explore the potential mechanisms of ZGW in promoting the osteogenesis of bone marrow mesenchymal stem cells (BMSCs). In total, 487 no-repeat targets corresponding to the bioactive components of ZGW were screened, and 175 target genes in the intersection of ZGW and osteogenesis were obtained. And 28 core target genes were then obtained from a PPI network analysis. A GO functional enrichment analysis showed that the relevant biological processes mainly involve the cellular response to chemical stress, metal ions, and lipopolysaccharide. Additionally, KEGG pathway enrichment analysis revealed that multiple signaling pathways, including the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) signaling pathway, were associated with ZGW-promoted osteogensis. Further experimental validation showed that ZGW could increase alkaline phosphatase (ALP) activity as well as the mRNA and protein levels of ALP, osteocalcin (OCN), and runt related transcription factor 2 (Runx 2). What's more, Western blot analysis results showed that ZGW significantly increased the protein levels of p-PI3K and p-AKT, and the increases of these protein levels significantly receded after the addition of the PI3K inhibitor LY294002. Finally, the upregulated osteogenic-related indicators were also suppressed by the addition of LY294002. ZGW promotes the osteogenesis of BMSCs via PI3K/AKT signaling pathway.

THU462 Clinical Features & Management Of FGF23 Secreting Tumors: Series Of 40 Clinical Cases

Abstract Disclosure: S. Gronskaia: None. Z. Belaya: None. L. Rozhinskaya: None. T. Dubovitskaya: None. E. Mamedova: None. E.A. Pigarova: None. M. Degtyarev: None. S. Rodionova: None. Y. Buklemishev: None. D. Babaeva: None. V. Vladimirova: None. N. Tarbaeva: None. S. Serzhenko: None. A. Grigoriev: None. L. Dzeranova: None. V. Karpenko: None. A. Karasev: None. R. Fedotov: None. I. Ulyanova: None. D. Trukhina: None. N. Toroptsova: None. O. Lesnyak: None. N.G. Mokrysheva: None. G.A. Melnichenko: None. I. Dedov: None. Introduction: Tumor-induced osteomalacia is an acquired rare disease manifested by hypophosphatemic osteomalacia due to excessive secretion of fibroblast growth factor 23 (FGF23). FGF 23 is a non-classical hormone secreted by bone tissue (osteocytes) and regulates phosphorus metabolism.The aim of this work is to present clinical experience in the diagnosis, treatment and rehabilitation of patients with tumor-induced osteomalacia. Materials and methods: 40 patients with clinically-confirmed tumor-induced osteomalacia were included in the study, 34 of whom had the tumor localized, 27 underwent surgical treatment and 21 achieved stable remission. Results: The average age was 48 +/- 14 years, 43% were men, the time left from the the onset of the disease was 8 +/- 6 years. Biochemical fundings were hypophosphatemia 0.47 [0.4 ; 0.53] mmol/l, a decrease in the tubular reabsorption phosphate 62 [52 ; 67]%, and an increase in alkaline phosphatase of 183 [112 ; 294] units/l. At the time of diagnosis, 100% had multiple pathological fractures, only 10% could move independently, and 76.3% classified the pain as unbearable (8-10 points according to the 10-point pain syndrome scale ). Among the methods used to detect tumors, the most sensitive were scintigraphy with tectrotide with SPECT/CT 71.4% (20/28) and MRI 90% (18/20). In 55% of cases, the tumor was localized in soft tissues and in 45% in bone tissue; The tumor was most often detected in the lower extremities, followed by the head in frequency of localization. 18 patients currently have no remission and they receive conservative treatment (phosphorus and alfacalcidol n=15 and burozumab n=3). In case of achieving remission (n=21), regression of clinical symptoms and restoration of bone and muscle mass was observed. Extensive excision of the tumor without prior biopsy resulted in the best percentage of remission - 87%. Conclusion: Tumor-induced osteomalacia is most common in the middle age people. It is characterized by severe damage to bone and muscle tissue with the development of multiple fractures, muscle weakness and severe pain syndrome. In laboratory diagnostics, attention should be paid to hypophosphatemia, a decrease in the tubutar reabsorption phosphate index and increased alkaline phosphatase. The use of functional diagnostic methods with a labeled somatostatin analogue to the subtype 2 receptor and MRI with contrast enhancement are the most accurate methods of topical diagnostics. In case of localization of the tumor, a wide excision without a preliminary biopsy is recommended. Presentation: Thursday, June 15, 2023

Development of a Novel Marine-Derived Tricomposite Biomaterial for Bone Regeneration.

Bone tissue engineering is a promising treatment for bone loss that requires a combination of porous scaffold and osteogenic cells. The aim of this study was to evaluate and develop a tricomposite, biomimetic scaffold consisting of marine-derived biomaterials, namely, chitosan and fucoidan with hydroxyapatite (HA). The effects of chitosan, fucoidan and HA individually and in combination on the proliferation and differentiation of human mesenchymal stem cells (MSCs) were investigated. According to the SEM results, the tricomposite scaffold had a uniform porous structure, which is a key requirement for cell migration, proliferation and vascularisation. The presence of HA and fucoidan in the chitosan tricomposite scaffold was confirmed using FTIR, which showed a slight decrease in porosity and an increase in the density of the tricomposite scaffold compared to other formulations. Fucoidan was found to inhibit cell proliferation at higher concentrations and at earlier time points when applied as a single treatment, but this effect was lost at later time points. Similar results were observed with HA alone. However, both HA and fucoidan increased MSC mineralisation as measured by calcium deposition. Differentiation was significantly enhanced in MSCs cultured on the tricomposite, with increased alkaline phosphatase activity on days 17 and 25. In conclusion, the tricomposite is biocompatible, promotes osteogenesis, and has the structural and compositional properties required of a scaffold for bone tissue engineering. This biomaterial could provide an effective treatment for small bone defects as an alternative to autografts or be the basis for cell attachment and differentiation in ex vivo bone tissue engineering.

Intervention Effects of Deer-Tendon Collagen Hydrolysates on Osteoporosis In Vitro and In Vivo.

Deer tendon, a deer processing byproduct, is an excellent protein source for the preparation of peptides for improving osteoporosis by its high protein content and high nutritional value. The optimal process of collagen acid extraction was implemented and the results showed that the acid concentration was 7%, the material-liquid ratio was 1:25, and the soaking time was 48 h. DTCHs could promote MC3T3-E1 cell proliferation and increase alkaline phosphatase activities in vitro. In addition, compared with the model group, the DTCHs treatment groups with an oral dosage of 350, 750, and 1500 mg/kg rat/day could significantly improve the shape, weight, bone mechanics, and alkaline phosphatase activities of tail-suspended mice. Bone microstructure and mineralization also recovered significantly in vivo. This result is expected to provide the structural and biological information for DTCHs-based functional foods.

Hepatic adenoma in a 7-year-old girl: a case report and literature review

BackgroundHepatocellular adenomas (HCAs) are rare benign tumors of the liver that occur predominantly in women taking oral contraceptives. In children, HCAs comprise < 5% of hepatic tumors. We report a case of HCAs in a 7-year-old girl with estrogen and glucose imbalance.Case presentationA 7-year-old girl was presented to our hospital with bilateral breast enlargement for 2 months, polydipsia, polyuria, polyphagia, hyperglycemia, and significant weight gain. Computed tomography (CT) showed a 7.2 cm×6.9 cm×5.3 cm round-shaped mass in the left inner lobe of the liver, ovarian ultrasound showed multiple follicles in the ovaries bilaterally, and cranial magnetic resonance imaging (MRI) showed an enlarged superior pituitary. Hematological and biochemical results were as follows: fasting glucose was 19.7 mmol/L, estradiol was 122.9 pmol/L, follicle-stimulating hormone 10.81 IU/L, luteinizing hormone 10.99 IU/L, insulin-like growth factor 1,513 ng/mL, glutamine aminotransferase 86 U/L, and alkaline phosphatase 362 U/L. Thyroid functions, methemoglobin, fetal protein, carcinoembryonic antigen, and chorionic gonadotropin were normal. The patient had a complete surgical resection of the liver tumor, and the postoperative histopathological diagnosis was HCAs. After the surgery, insulin was injected and the glucose levels were stable. During the 36-month follow-up period, neither tumor recurrence nor significant abnormalities were detected using color Doppler ultrasound of the liver. The child’s precocious puberty is currently under control.ConclusionsHCAs are particularly rare in children with liver tumors, and risk factors for the development of HCAs in children include sex hormone imbalance, obesity, Fanconi anemia (FA), glycogen storage diseases (GSDs) type I, III, and IV, galactosemia, immunodeficiency, congenital portosystemic shunts (CPSS), cardiac hepatopathy status-post Fontan procedure, Hurler syndrome, familial adenomatous polyposis, germline HNF1A mutations, and maturity-onset diabetes of the young type 3. Most HCAs are detected during a physical examination without clinical symptoms, and some patients may present with symptoms such as abdominal pain, abdominal distension, and abdominal masse. Serum liver function tests can show increased alkaline phosphatase (ALP) and γ- glutamyl transferase (GT), whereas α-Fetoprofein (AFP) levels are normal. The definitive diagnosis relies mainly on histopathological examination. Because HCAs can rupture and bleed and become malignant. Early surgical treatment is recommended after detection.

Effect of N-phenylbenzenesulfonamide targeting CNR2 over human bone marrow mesenchymal stem cell population and osteoblast/adipocyte differentiation

Cannabinoids receptor type 2 (CNR2) signaling using selective receptor compounds encourages the expression of osteogenic genes and enhances mineralization. The effect of six newly synthesized Cannabinoid receptors type 2 (CB2) selective compounds over the viability of human bone marrow mesenchymal stem cells (h-BMMSCs) and their osteogenic and adipogenic potential study was carried out. Human mesenchymal stem cells (MSCs) were treated with 0.5 μM of compounds 1–6 to induce lineage differentiation (adipogenic, osteogenic). The cells were analyzed using differentiation assays, lipid droplets and mineralized matrix was stained with “Niel red and Alizarin Red, further performed reverse transcription-polymerase chain reaction (RT-PCR) for lineage-specific markers expression.” Treatment with synthesized compounds N-(4-(dimethyl amino) benzyl)− 4-methyl-N-phenylbenzenesulfonamide (1), N-(4-(dimethyl amino) benzyl)-N-phenyl benzamide (3) and N-(4-tert-butylbenzyl)− 4-methyl-N-phenylbenzenesulfonamide (5), increased both lipid droplets formation and Alkaline Phosphatase (ALP) activity. Treatment of N-(4-(dimethylamino) benzyl)-N-phenylbenzene sulfonamide (2), and 4-methyl-N-phenyl-N-(4-(trifluoromethyl) benzyl) benzene sulfonamide (4) was found to enhance the adipogenic differentiation of (h-BMSCs) and inhibit osteogenic differentiation. A Reduction in adipocyte differentiation upon exposure to N-(4-tert-butylbenzyl)-N-phenylbenzene sulfonamide (6) was noted. Conversely, enhanced mineral deposition was accompanied by increased alkaline phosphatase (ALP) activity in osteogenic differentiation. Taken together, these newly synthesized selective compounds work in favor of adipocyte formation except for 6 which reduces lipid droplets accumulation and enhanced calcium deposition. “In conclusion, compounds 1, 3, 5 and 6 demonstrated a positive effect over osteoblast formation compared to control.” As per molecular docking study, compounds 1, 2 and 3 are the most active derivative will serve as anti-osteoporosis drug discovery.

Reed restoration decreased nutrients in wetlands with dredged sediments: Microbial community assembly and function in rhizosphere

Using dredged sediments as substrate for aquatic plants is a low-cost and ecological friendly way for in situ aquatic ecological restoration. However, the limited information available about how aquatic plant restoration affects the microbial ecology and nutrients in dredged sediments. In this study, nutrient contents, enzyme activities, and bacterial and archaeal communities in vertical sediment layers were determined in bulk and reed zones of wetlands constructed with dredged sediments in west Lake Taihu for three years. Reed restoration significantly decreased total nitrogen, total phosphorus, and organic carbon contents and increased alkaline phosphatase, urease, and sucrase activities compared to bulk area. Bacterial communities in vertical sediment layers had higher similarity in reed zone in comparison to bulk zone, and many bacterial and archaeal genera were only detected in reed rhizosphere zones. Compared with the bulk zone, the reed restoration area has a higher abundance of phylum Actinobacteriota, Hydrothermarchaeota, and class α-proteobacteria. The assembly process of the bacterial and archaeal communities was primarily shaped by dispersal limitation (67.03% and 32.97%, respectively), and stochastic processes were enhanced in the reed recovery area. Network analysis show that there were more complicated interactions among bacteria and archaea and low-abundance taxa were crucial in maintaining the microbial community stability in rhizosphere of reed zone. PICRUST2 analysis demonstrate that reed restoration promotes metabolic pathways related to C and N cycle in dredged sediments. These data highlight that using dredged sediments as substrates for aquatic plants can transform waste material into a valuable resource, enhancing the benefits to the environment.

Enhanced osteogenic and antibacterial properties of polyetheretherketone by ultraviolet-initiated grafting polymerization of a gelatin methacryloyl/epsilon-poly-L-lysine/laponite hydrogel coating.

Polyetheretherketone (PEEK) is a promising material for use in orthopedic implants, but its bio-inert character and lack of antibacterial activity limit its applications in bone repair. In the present study, considering the advantages of PEEK in self-initiated graft polymerization and of hydrogels in bone tissue engineering, we constructed a hydrogel coating (GPL) consisting of Gelatin methacryloyl (GelMA), methacrylamide-modified ε-poly-l-lysine (ε-PLMA) and Laponite on PEEK through UV-initiated crosslinking. The coating improved the hydrophilicity of PEEK, and the coating degraded slowly so that approximately 80% was retained after incubation in PBS for 8 weeks. In vitro studies revealed that as compared to culturing on PEEK, culturing on PEEK-GPL led to enhanced viability and adhesion of cultured human umbilical cord Wharton's jelly-derived mesenchymal stem cells (hWJ-MSCs). Due to the synergistic effect of the micron-scale three-dimensional surface and Laponite, PEEK-GPL exhibited a significantly improved induction of osteogenic differentiation of hWJ-MSCs compared to PEEK, as demonstrated by increased alkaline phosphatase activity, matrix mineralization, and expression of osteogenesis-related genes. Furthermore, PEEK-GPL showed antibacterial activity upon contact with Staphylococcus aureus and Escherichia coli, and this activity would be maintained before complete degradation of the hydrogel because the ε-PLMA was cross-linked covalently into the coating. Thus, PEEK-GPL achieved both osteogenesis and infection prevention in a single simple step, providing a feasible approach for the extensive use of PEEK in bone implants.

Kaempferol-loaded bioactive glass-based scaffold for bone tissue engineering: in vitro and in vivo evaluation

Due to the increasing prevalence of bone disorders among people especially in average age, the future of treatments for osseous abnormalities has been illuminated by scaffold-based bone tissue engineering. In this study, in vitro and in vivo properties of 58S bioactive glass-based scaffolds for bone tissue engineering (bare (B.SC), Zein-coated (C.SC), and Zein-coated containing Kaempferol (KC.SC)) were evaluated. This is a follow-up study on our previously published paper, where we synthesized 58S bioactive glass-based scaffolds coated with Kaempferol-loaded Zein biopolymer, and characterized from mostly engineering points of view to find the optimum composition. For this aim, in vitro assessments were done to evaluate the osteogenic capacity and biological features of the scaffolds. In the in vivo section, all types of scaffolds with/without bone marrow-derived stem cells (BMSC) were implanted into rat calvaria bone defects, and potential of bone healing was assessed using imaging, staining, and histomorphometric analyses. It was shown that, Zein-coating covered surface cracks leading to better mechanical properties without negative effect on bioactivity and cell attachment. Also, BMSC differentiation proved that the presence of Kaempferol caused higher calcium deposition, increased alkaline phosphatase activity, bone-specific gene upregulation in vitro. Further, in vivo study confirmed positive effect of BMSC-loaded KC.SC on significant new bone formation resulting in complete bone regeneration. Combining physical properties of coated scaffolds with the osteogenic effect of Kaempferol and BMSCs could represent a new strategy for bone regeneration and provide a more effective approach to repairing critical-sized bone defects.

Effect of Subsurface Drainage Combined with Biochar on the Bacterial Community Composition of Coastal Saline Soil

Waterlogging and salinization are considered to be the main threats to agricultural productivity and land resources in coastal areas of China. Thus far, drainage and field soil improvement programs have been ineffective. In this article, we investigated the effect of subsurface drainage combined with biochar (B–S) on soil physicochemical properties and soil bacterial community structure in coastal saline soil. In this study, B–S significantly reduced soil electrical conductivity (EC) and soil water content (W) by 35% and 10.65% compared to no drainage (CK). Compared to CK and drainage alone (S), B–S significantly increased soil total nitrogen (TN) by 24.78% and 39.62%, soil available phosphorus (AP) by 28.29% and 69.82%, soil nitrate (NO3−-N) by 64.65% and 35.45%, and significantly increased soil organic matter (SOM) by 74.69% and 66.10%, respectively. It also significantly increased alkaline phosphatase (ALP) and urease activities. The results of redundancy analysis (RDA) showed that CAT and urease made the greatest response to changes in environmental factors, indicating that CAT is more sensitive to changes in environmental alterations than ALP. AP was the dominant factor in the change in enzyme activity (R2 = 53.0%, p &lt; 0.05), followed by NO3−-N (R2 = 14.8%). SOM was the dominant factor in the variation in microbial abundance content (R2 = 38.5%, p &lt; 0.05), followed by ALP (R2 = 20.0%, p &lt; 0.05). The results of the study can provide guidance for effective land use and sustainable development of agricultural soil ecology in coastal areas.

Effects of Dietary Vitamin D3 Over-Supplementation on Broiler Chickens' Health; Clinicopathological and Immunohistochemical Characteristics

Introduction: Vitamin D3 is used as a supplement in the feeds of livestock, pets, and human infants. However, the presence of excessive Vitamin D3 has been shown to cause toxicity in humans and animals. This study investigated the clinicopathological aspects of Vitamin D3 toxicity in broiler chickens. Materials and methods: The median lethal dose (LD50) of Vitamin D3 was estimated by the up-and-down method. To determine long term (21 days) toxic effects of oral Vitamin D3 supplementation, 90 (14-day-old) IBL-80 unsexed chicks were randomly divided into three groups as group A (control, received basal diet), B (basal diet + Vitamin D3 at 16.67 mg/kg body weight daily), and C (basal diet + Vitamin D3 at 33.33 mg/kg body weight daily). Results: The findings indicated that broiler chickens tolerated a single oral dose of Vitamin D3 up to 550 mg/kg body weight (22,000,000 IU/kg) without mortality. The results of long-term (21 days) oral supplementation of divided doses of Vitamin D3 in broiler chickens (groups B and C) showed progressive emaciation, elevated hemoglobin, hypercalcemia, hypophosphatemia, and increased alkaline phosphatase activity. At necropsy, pale liver and kidneys, congestion and hardening of lungs, mild congestion in the brain, and soft bones were observed in Vitamin D3 treated chicks (groups B and C). Microscopically, degeneration and metastatic calcification in lung parenchyma and peribronchiolar epithelium, coagulative necrosis and calcification in kidneys, and calcification with fibroplasia in proventriculus was detected. Lungs and kidneys showed a significant difference in calcification score between groups B and C. Broiler chickens from Vitamin D3 treated groups (B and C) showed strong immunohistochemical expression of Calbindin D28K in the intestine and kidneys but weak expression in the lungs. Conclusion: This study demonstrates that broiler chicks can tolerate very high levels of a single oral dose of Vitamin D3. Toxic effects of prolonged exposure to Vitamin D3 are due to over-expression of Calbindin D28k in the intestine and kidneys, disturbing the calcium and phosphorous homeostasis and leading to metastatic calcification of vital organs. This study supports that prolonged over-supplementation of Vitamin D3 causes toxic effects; hence, appropriate dietary Vitamin D3 supplementation limits should be set.

Combined effects of ocean deoxygenation, acidification, and phosphorus limitation on green tide macroalga, Ulva prolifera

Ocean deoxygenation, acidification, and decreased phosphorus availability are predicted to increase in coastal ecosystems under future climate change. However, little is known regarding the combined effects of such environmental variables on the green tide macroalga Ulva prolifera. Here, we provide quantitative and mechanistic understanding of the acclimation mechanisms of U. prolifera to ocean deoxygenation, acidification, and phosphorus limitation under both laboratory and semi-natural (mesocosms) conditions. We found that there were significant interactions between these global environmental conditions on algal physiological performance. Although algal growth rate and photosynthesis reduced when the nitrogen-to‑phosphorus (N/P) ratio increased from 16:1 to 35:1 under ambient CO2 and O2 condition, they remained constant with further increasing N/P ratios of 105:1, 350:1, and 1050:1. However, the increasing alkaline phosphatase activities at high N/P ratios suggests that U. prolifera could use organic P to support its growth under phosphorus limitation. Deoxygenation had no effect on specific growth rate (SGR) but decreased photosynthesis under low N/P ratios of 16:1, 35:1, and 105:1, with reduced activities of several enzymes involved in N assimilation pathway being observed. Elevated CO2 promoted algal growth and alleviated the negative effect of deoxygenation on algal photosynthesis. The patterns of responses to high CO2 and low O2 treatments in in situ experiments were generally consistent with those observed in laboratory experiments. Our results generally found that the strong physiological acclimation capacity to elevated CO2, low O2, and high N/P could contribute to its large-scale blooming in coastal ecosystem.

Effects of Gamma Irradiation and Supercritical Carbon Dioxide Sterilization on Methacrylated Gelatin/Hyaluronan Hydrogels.

Biopolymer hydrogels have become an important group of biomaterials in experimental and clinical use. However, unlike metallic or mineral materials, they are quite sensitive to sterilization. The aim of this study was to compare the effects of gamma irradiation and supercritical carbon dioxide (scCO2) treatment on the physicochemical properties of different hyaluronan (HA)- and/or gelatin (GEL)-based hydrogels and the cellular response of human bone marrow-derived mesenchymal stem cells (hBMSC). Hydrogels were photo-polymerized from methacrylated HA, methacrylated GEL, or a mixture of GEL/HA. The composition and sterilization methods altered the dissolution behavior of the biopolymeric hydrogels. There were no significant differences in methacrylated GEL release but increased methacrylated HA degradation of gamma-irradiated samples. Pore size/form remained unchanged, while gamma irradiation decreased the elastic modulus from about 29 kPa to 19 kPa compared to aseptic samples. HBMSC proliferated and increased alkaline phosphatase activity (ALP) particularly in aseptic and gamma-irradiated methacrylated GEL/HA hydrogels alike, while scCO2 treatment had a negative effect on both proliferation and osteogenic differentiation. Thus, gamma-irradiated methacrylated GEL/HA hydrogels are a promising base for multi-component bone substitute materials.

Dissecting specific Wnt components governing osteogenic differentiation potential by human periodontal ligament stem cells through interleukin-6

Periodontal ligament stem cells (PDLSCs) play a significant role on periodontal tissue and alveolar bone homeostasis. During inflammation, interleukin (IL)-6 serves as one of key cytokine players controlling tissue reaction as well as alveolar bone tissue remodeling. It is believed that periodontal tissue inflammation causes periodontium degradation, especially alveolar bone. However, in this study, we show that an inflammatory mediator, IL-6, may serve another direction on alveolar bone homeostasis during inflammatory condition. We found that, IL-6 at 10 and 20 ng/mL was not cytotoxic and dose-dependently exerted beneficial effects on osteogenic differentiation of human PDLSCs (hPDLSCs), as demonstrated by increased alkaline phosphatase activity, mRNA expression of osteogenic markers, and matrix mineralization. The presence of physiological and inflammatory level of IL-6, the osteogenic differentiation potential by hPDLSCs was enhanced by several possible mechanisms including transforming growth factor (TGF), Wnt, and Notch pathways. After in-depth and thorough exploration, we found that Wnt pathway serves as key regulator controlling osteogenic differentiation by hPDLSCs amid the IL-6 presentation. Surprisingly, apart from other mesenchymal stem cells, distinct Wnt components are employed by hPDLSCs, and both canonical and non-canonical Wnt pathways are triggered by different mechanisms. Further validation by gene silencing, treatment with recombinant Wnt ligands, and β-catenin stabilization/translocation confirmed that IL-6 governed the canonical Wnt/β-catenin pathway via either WNT2B or WNT10B and employed WNT5A to activate the non-canonical Wnt pathway. These findings fulfill the homeostasis pathway governing periodontal tissue and alveolar bone regeneration and may serve for further therapeutic regimen design for restoring the tissues.

Phase I Beamion Lung 1 trial of BI 1810631, a HER2 tyrosine kinase inhibitor (TKI), as monotherapy in patients (pts) with advanced/metastatic solid tumors with HER2 aberrations: Updated data.

8545 Background: There is an unmet need for effective TKIs against HER2 mutations in solid tumors, especially NSCLC. BI 1810631 is a highly potent and selective HER2 TKI that covalently binds to both wild-type and mutated HER2 receptors, including exon 20 insertions (ex20ins), while sparing EGFR. Beamion Lung 1 is an ongoing Phase Ia/Ib study to determine the safety, MTD, PK, PD, and preliminary efficacy of BI 1810631 in pts with HER2 aberration-positive solid tumors (NCT04886804). Preliminary results from Phase Ia are presented. Methods: In Phase Ia, pts with advanced/unresectable/metastatic solid tumors who were refractory or unsuitable for standard treatment (Tx) and had HER2 aberrations (e.g. overexpression, gene amplification, somatic mutation, or gene rearrangements) were enrolled. Pts received escalating doses of BI 1810631 BID starting from 15 mg or BI 1810631 QD starting from 60 mg via a Bayesian model. Phase Ib will initially include 30 pts with advanced HER2 tyrosine kinase domain mutation-positive, pre-treated NSCLC. Additional cohorts may be included. Primary endpoints: MTD based on DLTs; pts with DLTs (Phase Ia); ORR (Phase Ib). Secondary endpoints: number of pts with DLTs throughout entire Tx period and PK parameters (Phase Ia/Ib); DoR, DCR, duration of disease control, and PFS (Phase Ib). Results: As of 23 January 2023, 36 pts had been treated in the US, The Netherlands, Japan, and China. Pts had NSCLC (n = 22), colorectal cancer (n = 3), or other tumors (n = 11). Most pts (n = 26; ex20ins: n = 23) had a pathological HER2 mutation. Median (range) lines of prior systemic Tx: 2.5 (1–8). Prior HER2-targeted Tx: 12 (33%). Pts received BI 1810631 at 15, 30, 60, 100, 150 mg BID (n = 3/3/4/4/3) or 60, 120, 180, 240, 300 mg QD (n = 5/4/5/4/1). Median number of cycles was 4 (range 1–14). Tx is ongoing in 20 pts. To date, 3 DLTs have been observed (grade 2 edema [60 mg BID], grade 3 anemia [60 mg QD], grade 3 elevated ALT [180 mg QD]). The MTD has not been reached with either schedule. Tx-related adverse events (TRAEs) were reported in 25 pts (69%). The most common TRAEs were diarrhea (n = 10), anemia (n = 5), increased alkaline phosphatase, increased creatinine, increased ALT, hypoalbuminemia (all n = 4), hypocalcemia, increased AST, dry skin, increased GGT (all n = 3). Three pts had grade 3 TRAEs (anemia/increased GGT [n = 1], increased ALT [n = 2]). Strong preliminary efficacy signals were observed across all dose levels. Of 22 evaluable NSCLC patients, 10 responded (all PRs; all ex20ins) and 11 had SD. Two patients with other tumors (esophagus; cholangiocarcinoma) also achieved PR. Conclusions: These preliminary data indicate that BI 1810631 is well tolerated and shows encouraging anti-tumor activity in pts with HER2 aberration-positive solid tumors. Recruitment into Phase Ia is ongoing. Updated data, including durability endpoints, will be presented. Clinical trial information: NCT04886804 .