ALP Activity Research Articles

Material extrusion additive manufacturing of poly(lactic acid)/Ti6Al4V@calcium phosphate core-shell nanocomposite scaffolds for bone tissue applications

The use of porous scaffolds with appropriate mechanical and biological features for the host tissue is one of the challenges in repairing critical-size bone defects. With today's three-dimensional (3D) printing technology, scaffolds can be customized and personalized, thereby eliminating the problems associated with conventional methods. In this work, after preparing Ti6Al4V/Calcium phosphate (Ti64@CaP) core-shell nanocomposite via a solution-based process, by taking advantage of fused deposition modeling (FDM), porous poly(lactic acid) (PLA)-Ti64@CaP nanocomposite scaffolds were fabricated. Scanning electron microscope (SEM) showed that nanostructured calcium phosphate was distributed uniformly on the surface of Ti64 particles. Also, X-ray diffraction (XRD) indicated that calcium phosphate forms an octacalcium phosphate (OCP) phase. As a result of incorporating 6 wt% Ti64@CaP into the PLA, the compressive modulus and ultimate compressive strength values increased from 1.4 GPa and 29.5 MPa to 2.0 GPa and 53.5 MPa, respectively. Furthermore, the differential scanning calorimetry results revealed an increase in the glass transition temperature of PLA, rising from 57.0 to 62.4 °C, due to the addition of 6 wt% Ti64@CaP. However, it is worth noting that there was a moderate decrease in the crystallization and melting temperatures of the nanocomposite filament, which dropped from 97.0 to 89.5 °C and 167 to 162.9 °C, respectively. The scaffolds were seeded with human adipose tissue-derived mesenchymal stem cells (hADSCs) to investigate their biocompatibility and cell proliferation. Calcium deposition, ALP activity, and bone-related proteins and genes were also used to evaluate the bone differentiation potential of hADSCs. The obtained results showed that introducing Ti64@CaP considerably improved in vitro biocompatibility, facilitating the attachment, differentiation, and proliferation of hADSCs. Considering the findings of this study, the 3D-printed nanocomposite scaffold could be considered a promising candidate for bone tissue engineering applications.

Association between dietary selenium intake and the prevalence of osteoporosis and its role in the treatment of glucocorticoid-induced osteoporosis

BackgroundLong-term glucocorticoid therapy may lead to osteoporosis (OP). Selenium (Se) is an essential microelement for human health and bone health. This study evaluated the association between dietary Se intake and the prevalence of OP and further explored the potential therapeutic effect of Se on glucocorticoid-induced OP (GIOP) in vivo and in vitro.MethodsData were collected from a population-based cross-sectional study conducted in our hospital. OP is diagnosed based on bone mineral density (BMD) measurements using compact radiographic absorptiometry. Dietary Se intake was assessed using a semi-quantitative food frequency questionnaire. The association between dietary Se intake and OP prevalence was analyzed by multivariable logistic regression. In animal experiments, male Sprague–Dawley rats were intramuscularly injected with dexamethasone (1 mg/kg) daily to induce GIOP, while different doses of Se were supplemented in rat drinking water for 60 d. BMD and biomechanical parameters of rat femur were measured. The histopathological changes of the femur were observed by HE staining, the number of osteoclasts was observed by TRAP staining, and OCN positive expression was detected by immunohistochemical staining. OPG, RANKL, Runx2, and BMP2 in rat femur were detected by Western blot. Bone turnover markers and oxidative stress markers were measured using commercial kits. MC3T3-E1 cells were induced to osteogenic differentiation, stimulated with DXM (100 μM), and/or treated with Se at different doses. Cell proliferation and apoptosis were assessed by CCK-8 and flow cytometry. ALP activity was detected by ALP staining and cell mineralization was observed by alizarin red staining.ResultsParticipants with lower dietary Se intake had higher OP prevalence. Se supplementation improved BMD, biomechanical parameters, and histopathological changes of the femur in GIOP rats. Se supplementation also suppressed DXM-induced changes in bone turnover- and oxidative stress-related markers. Under DXM conditions, Se treatment induced MC3T3-E1 cell proliferation, ALP activity, and mineralization.ConclusionLower Dietary Se intake is associated with OP prevalence. Moreover, Se takes a position in bone protection and anti-oxidative stress in GIOP models. Therefore, Se may be a complementary potential treatment for GIOP.

Enhancement of in vitro antibacterial activity and bioactivity of iodine-loaded titanium by micro-scale regulation using mixed-acid treatment.

Postoperative infection and subsequent device loss are serious complications in the use of titanium dental implants and plates for jawbone reconstruction. We have previously reported that NaOH-CaCl2 -thermal-ICl3 -treated titanium (NaCaThIo) has a nano-scale surface and exhibits antibacterial activity against Staphylococcus aureus. The present study examined the surface properties of mixed-acid treated and then iodine-treated titanium (MA-NaCaThIo), and evaluated oral antibacterial activity and cytotoxicity compared with the results obtained with NaCaThIo. MA-NaCaThIo formed a surface layer with a nano-scale network structure having microscale irregularities, and both the thickness of the surface layer (1.49 ± 0.16 μm) and the average surface roughness (0.35 ± 0.03 μm) were significantly higher than those of NaCaThIo. Furthermore, MA-NaCaThIo maintained high hydrophilicity with a contact angle of 7.5 ± 1.7° even after 4 weeks, as well as improved apatite formation, iodine ion release, and antibacterial activity against Prevotella intermedia compared to NaCaThIo. Cell culture test revealed that MA-NaCaThIo exhibited no cytotoxicity against MG-63 and Vero cells, while increased cell proliferation, ALP activity and mineralization of MG-63 compared to NaCaThIo. This treated titanium is expected to be useful for the development of next-generation titanium devices having both bone-bonding and antibacterial properties.

Gallic acid induces osteoblast differentiation and alleviates inflammatory response through GPR35/GSK3β/β-catenin signaling pathway in human periodontal ligament cells.

Gallic acid (GA) possesses various beneficial functions including antioxidant, anticancer, anti-inflammatory as well as inhibiting osteoclastogeneis. However, effects on osteogenic differentiation, especially in human ligament periodontal (hPDL) cells, remain unclear. Thus, the aim of this study was to evaluate the function of GA on osteogenesis and anti-inflammation in hPDL cells and to explore the involved underlying mechanism. Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) treatment was used as a model for periodontitis. ROS production was determined by H2DCFDA staining. Trans-well and wound healing assays were performed for checking the migration effect of GA. Alizarin red and alkaline phosphatase activity (ALP) assays were performed to evaluate osteogenic differentiation. Osteogenesis and inflammatory-related genes and proteins were measured by real-time PCR and western blot. Our results showed that GA-treated hPDL cells had higher proliferation and migration effect. GA inhibited ROS production-induced by Pg-LPS. Besides, GA abolished Pg-LPS-induced inflammation cytokines (il-6, il-1β) and inflammasome targets (Caspase-1, NLRP3). In addition, GA promoted ALP activity and mineralization in hPDL cells, lead to enhance osteoblast differentiation process. The effect of GA is related to G-protein-coupled receptor 35 (GPR35)/GSK3β/β-catenin signaling pathway. GA attenuated Pg-LPS-induced inflammatory responses and periodontitis in hPDL cells. Taken together, GA may be targeted for therapeutic interventions in periodontal diseases.

LncRNA JHDM1D-AS1 promotes osteogenic differentiation of periodontal ligament cells by targeting miR-532-5p to activate IGF1R signaling.

The aim of this study was to explore the mechanism underlying periodontal ligament cells (PDLCs) osteogenic differentiation. Periodontitis causes damage to tooth-supporting apparatus and eventually leads to tooth loss. PDLCs hold great promise in periodontal regeneration due to their osteogenic features. The expression of osteogenic markers, lncRNA JHDM1D-AS1, miR-532-5p and IGF1R was examined. For osteogenic differentiation, primary human PDLCs (hPDLCs) were cultured in an osteogenic medium, and it was assessed by ALP activity and Alizarin Red staining. The interaction between JHDM1D-AS1, miR-532-5p and IGF1R was analyzed via dual luciferase, RIP and RNA pull-down assays. JHDM1D-AS1 was up-regulated during osteogenic differentiation and its silencing inhibited hPDLC osteogenic differentiation. JHDM1D-AS1 worked as a miR-532-5p sponge in hPDLCs. miR-532-5p directly targeted IGF1R to suppress its expression, and miR-532-5p knockdown facilitated osteogenic differentiation of hPDLCs. Overexpression of IGF1R promoted osteogenic differentiation of hPDLCs via activating Notch/HES1 signaling in hPDLCs. JHDM1D-AS1 promotes osteogenic differentiation of hPDLCs via sponging miR-532-5p to facilitate IGF1R expression and activate Notch/HES1 signaling.

A novel technique to harvest bone autografts with mild local hyperthermia and enhanced osteogenic bone quality: a preclinical study in dogs

BackgroundGuided bone regeneration (GBR) involves collecting bone autografts with high bio-quality and efficiency. The current non-irrigated low-speed drilling has been limited for broader application in bone autograft harvest due to its low efficiency, inability to conduct buccal cortical perforation, and dependence on simultaneous implant placement. Increasing the drilling speed helps improve the efficiency but may incur thermal-mechanical bone damage. Most studies have addressed thermal reactions during bone drilling on non-vital models, which is irrelevant to clinical scenarios. Little has been known about bone’s in vivo thermal profiles under non-irrigated higher-speed drilling and its influences on the resulting bone chips.AimA novel technique for bone harvest and cortical perforation via in-situ non-irrigated higher-speed drilling was proposed and investigated for the first time.MethodsThe third mandible premolars of eight beagles were extracted and healed for three months. Sixteen partial edentulous sites (left and right) were randomized into four groups for bone autograft harvest without irrigation: chisel, 50 rpm drilling, 500 rpm drilling, and 1000 rpm drilling. Bone chips were harvested on the buccal plates of the missing tooth. An infrared camera and an implantable thermocouple collaboratively monitored in vivo real-time bone temperature at the drilling sites. In vitro performances of cells from bone chips, including cell number, viability, proliferation, migration, ALP activity, in vitro mineralization, mRNA transcriptional level of osteogenic genes and heat shock protein 70 (HSP-70), and HSP-70 expression at the protein level were also studied.Results500 rpm produced mild local hyperthermia with a 2–6 °C temperature rise both on the cortical surface and inside the cortical bone. It also held comparable or enhanced cell performances such as cell number, viability, proliferation, migration, ALP activity, in vitro mineralization, and osteogenic genes expression.ConclusionsIn-situ non-irrigated higher-speed drilling at 500 rpm using a screw drill is versatile, efficient, and thermal friendly and improves the bio-quality of bone chips. Our novel technique holds clinical translational potential in GBR application.

Zizyphus mauritiana seed extract: Paving the way for next-generation bone constructs with nano-fluorohydroxyapatite/carboxymethyl chitosan nanocomposite scaffold

This is the first study that explored the potential use of Zizyphus mauritiana seed extract (ZSE) to synthesize nano-fluorohydroxyapatite/carboxymethyl chitosan nanocomposite scaffolds at different concentrations (CFZ1, CFZ2 and CFZ3) using co-precipitation method. The proposed scaffolds showed presence of intermolecular H bonding interactions between the constituents, according to the FTIR. The mechanical studies revealed shore hardness of 72 ± 4.6 and optimal compressive modulus in case of CFZ3 [1654.48 ± 1.6 MPa], that was comparable with that of human cortical bone. The SEM, TEM and platelet adhesion images corroborated uniformly distributed needle like particles in case of CFZ3 with an average size ranging from 22 to 26 nm, linked rough morphology, and appropriate hemocompatibility. The markedly up regulation in the ALP activity and protein adsorption upon increasing ZSE concentration demonstrated that CFZ nanocomposite scaffolds were compatible with osteoblastic cells relative to CF nanocomposite. The cytotoxicity study indicated that CFZ nanocomposite do not induce toxicity over MG-63 and did not aggravate LDH leakage in contrast to CF. The histopathological investigations on albino rats confirmed significantly improved regeneration of bone in the repair of a critical-size [8 mm] calvarium defect. Therefore, CFZ3 nanocomposite scaffold represents a simple, off-the-shelf solution to the combined challenges associated with bone defects.

Self-assembled chitosan/gelatin nanofibrous aggregates incorporated thermosensitive nanocomposite bioink for bone tissue engineering

Chitosan-based thermosensitive bioink can be a potential option as bioinks for bone tissue engineering because of their excellent biocompatibility and crosslinker-free gelation at physiological temperature. However, their low mechanical strength, poor printability, and low post-printing cell viability are some of their limitations. In this work, self-assembled nanofibrous aggregates of chitosan and gelatin were prepared and incorporated in chitosan-based bioinks to enhance printability, mechanical properties, post-printing cell viability, and proliferation. Subsequently, the optimal concentration of nanohydroxyapatite was determined, and the potential of the nanocomposite bioink was evaluated. Physiochemical, mechanical, and in vitro characterizations were carried out for the developed nanocomposite bioink. The bioink had optimum printability at 10 % nanohydroxyapatite and cell viability >88 %. The composite bioink had a low water uptake capacity (2.5 %) and degraded within 3 weeks in the presence of lysozyme. Mechanical characterization revealed an elastic modulus of about 15.5 kPa. Rheological analysis indicated a higher storage modulus of the bioink samples at 37 °C. ALP activity of 36.8 units/ml after 14 days of scaffold culture in osteogenic media indicated high cellular activity. These results suggested that the incorporation of osteogenic nanohydroxyapatite and nanofibrous aggregates improved the overall osteogenic and physiochemical potential of the thermosensitive bioink.

Preliminary evaluation of human hair follicle-derived dermal papilla cell characteristics

Dermal Papilla Cells (DPCs) are mesenchymal cells residing at the base of the hair follicles. They play an important role in the hair cycle remodeling and hair follicle regeneration. In this study, DPCs were isolated, and cultured from human hair follicles and evaluated the expression of -SMA, versican by immunocytochemistry, and alkaline phosphatase activity. The human fibroblasts were used as a control to confirm the non-fibroblast DPCs. The results showed that the cells grew from the hair bulb and had fibroblast morphology. These bulb cells strongly expressed alkaline phosphatase activity -SMA, and versican while the human fibroblast lacked ALP activity and less expression of -SMA and versican. In conclusion, the protocol is established to isolate and culture DPCs from hair follicle bulbs, and ALP activity is suggested as the major marker for distinguishing DPCs and human fibroblasts.

Effect of ethanolic leaf extracts of Medicago sativa, Gongronema latifolium and Pterocarpus sanalinoides on the liver integrity of alloxan-induced diabetic albino rats

The study investigated the effects of ethanolic leaf extracts of Medicago sativa (MS), Pterocarpus santalinoides (PS) and Gongronema latifolium (GL) on the liver function profile and organ histology integrity of alloxan-induced diabetic albino rats. Sixty male rats were divided into 10 groups, made up of 1 group each of normal, diabetic, and positive controls, 3 treatment groups each of the single and pairwise extract combinations, and 1 group of the ternary extracts combination. Hyperglycaemia was induced in the treatment groups by a single intraperitoneal injection of alloxan at 150 mg/kg body weight. The positive control was treated with metformin (100 mg/kg), while the extracts were administered via oral intubation to the single (200 mg/kg of each extract), double (combination of 2 extracts at 100 mg/kg each) and ternary (combination of the 3 extracts at 66.7 mg/kg each) treatment groups for a total period of 4 weeks. Induction of diabetes significantly (p<0.05) increased AST, ALT and ALP activities and total bilirubin concentration, but treatment with metformin and the plants’ extracts significantly (p<0.05) reduced the enzymes activities and total bilirubin concentration to levels comparable to those of the normal control. Generally, the combination therapies had non-significantly stronger effects than the single extract treatments. Histological evaluation confirmed that combinatorial compared to individual extracts treatment reverted more the damage to the liver organ caused by alloxan induction. Thus, Medicago sativa, Pterocarpus santalinoides and Gongronema latifolium are possible lead plants for potential production of hepatoprotective agents.

Microplastics and plastic additives as contaminants of emerging concern: A multi-biomarker approach using Rhinella arenarum tadpoles

Polyethylene microplastics (PE-MPs), a whitish thermoplastic polymer with numerous applications, is one of the materials most widely used in the industrial sector, whereas tetrabromobisphenol A (TBBPA) is a brominated flame retardant. The aim of this study was to analyze the effects of PE-MPs and TBBPA on Rhinella arenarum tadpoles at the laboratory scale. Tadpoles were chronically exposed (30 days) to four treatments: PE-MPs (60 mg L−1), TBBPA (10 µg L−1), their mixture (PE-MPs + TBBPA), and dechlorinated water as negative control (CO). Biomarkers of enzymatic activity (acetylcholinesterase, AChE; carboxylesterase, CbE; glutathione reductase, GR; and glutathione-S-transferase, GST), hepatic physiological alteration (alkaline phosphatase; ALP activity, and cholesterol; CHOL level), and endocrine disruption through thyroid hormone (T4) levels were assessed. In addition, intestine and liver were histomorphologically evaluated. AChE activity in tadpoles was significantly inhibited after exposure to PE-MPs and TBBPA with respect to CO. In addition, CbE, GR, and ALP activities showed higher values in the mixture of PE-MPs+ TBBPA treatment than in CO, whereas CHOL level was higher in TBBPA and PE-MPs+ TBBPA treatments than in CO. GST activity did not show significant differences between treatments and CO. T4 levels increased significantly in all treatments with respect to CO. The intestinal structure of tadpoles exposed to PE-MPs and PE-MPs+ TBBPA showed signs of mechanical damage. The intestinal wall of tadpoles under PE-MPs, TBBPA and PE-MPs+ TBBPA treatments was thicker than that of CO individuals. The analysis of liver histology demonstrated the hepatotoxicity caused by PE-MPs+ TBBPA. This study provides quantitative evidence of the harmful effects of PE-MPs, TBBPA and their mixture on enzymatic and hormonal activities, and histological evidence of intestinal wall hypertrophy and liver damage of R. arenarum tadpoles.

Mineralized Microgels via Electrohydrodynamic Atomization: Optimization and In Vitro Model for Dentin-Pulp Complex.

There is growing interest in the use of micro-sized hydrogels, including bioactive signals, as efficient platforms for tissue regeneration because they are able to mimic cell niche structure and selected functionalities. Herein, it is proposed to optimize bioactive composite microgels via electrohydrodynamic atomization (EHDA) to regenerate the dentin-pulp complex. The addition of disodium phosphate (Na2HPO4) salts as mineral precursors triggered an in situ reaction with divalent ions in solution, thus promoting the encapsulation of different amounts of apatite-like phases. Morphological analysis via image analysis of optical images confirmed a narrow distribution of perfectly rounded particles, with an average diameter ranging from 223 ± 18 μm to 502 ± 64 μm as a function of mineral content and process parameters used. FTIR, TEM, and EDAX analyses confirmed the formation of calcium phosphates with a characteristic Ca/P ratio close to 1.67 and a needle-like crystal shape. In vitro studies-using dental pulp stem cells (DPSCs) in crown sections of natural teeth slices-showed an increase in cell viability until 14 days, recording a decay of proliferation at 21 days, independent on the mineral amount, suggesting that differentiation is started, as confirmed by the increase of ALP activity at 14 days. In this view, mineralized microgels could be successfully used to support in vitro osteogenesis, working as an interesting model to study dental tissue regeneration.

Mechanical stress-induced FGF-2 promotes proliferation and consequently induces osteoblast differentiation in mesenchymal stem cells

Mechanical stimuli serve as crucial regulators of bone mass, promoting bone formation. However, the molecular mechanisms governing how mesenchymal stem cells (MSCs) respond to mechanical cues during their differentiation into osteogenic cells remain elusive. In this study, we found that cyclic stretching enhances MSC proliferation but does not increase the expression of osteoblast-related genes. We further revealed that this proliferative effect is mediated by fibroblast growth factor 2 (FGF-2), synthesized by MSCs in response to mechanical stress. Cell proliferation induced by cyclic stretching was inhibited upon the addition of either U0126, an inhibitor of mitogen-activated protein kinase kinase (MEK), or early growth response 1 (EGR1)-targeting small-hairpin RNA (shRNA), indicating the involvement of the extracellular signal-regulated kinase (ERK)/EGR1 signaling pathway. Osteoblast differentiation, evaluated through ALP activity, osteoblast-related gene expression, and mineralization, was stimulated by recombinant human FGF-2 (rhFGF-2) when applied during the proliferation phase, but not when applied during the differentiation stage alone. Our results suggest that FGF-2 indirectly promotes osteoblast differentiation as a downstream effect of stimulating cell proliferation. For the first time, we demonstrate that cyclic stretching induces MSCs to produce FGF-2, which in turn encourages cell proliferation through an autocrine/paracrine mechanism, consequently leading to osteoblast differentiation.

Evaluating osteogenic effects associated with the incorporation of ascorbic acid in mineralized collagen scaffolds.

Current treatments for craniomaxillofacial (CMF) defects motivate the design of instructive biomaterials that can promote osteogenic healing of complex bone defects. We report methods to promote in vitro osteogenesis of human mesenchymal stem cells (hMSCs) within a model mineralized collagen scaffold via the incorporation of ascorbic acid (vitamin C), a key factor in collagen biosynthesis and bone mineralization. An addition of 5 w/v% ascorbic acid into the base mineralized collagen scaffold significantly changes key morphology characteristics including porosity, macrostructure, and microstructure. This modification promotes hMSC metabolic activity, ALP activity, and hMSC-mediated deposition of calcium and phosphorous. Additionally, the incorporation of ascorbic acid influences osteogenic gene expression (BMP-2, RUNX2, COL1A2) and delays the expression of genes associated with osteoclast activity and bone resorption (OPN, CTSK), though it reduces the secretion of OPG. Together, these findings highlight ascorbic acid as a relevant component for mineralized collagen scaffold design to promote osteogenic differentiation and new bone formation for improved CMF outcomes.

Preparation, properties and in vitro osteogensis of self-reinforcing injectable hydrogel

As an attractive biomaterial for bone reconstruction, injectable biomaterials have many prominent characteristics such as good biocompatibility and bone-filling ability. However, there are weak as load-bearing scaffolds. In this study, polyvinyl alcohol (PVA) and bioactive glass (BAG) were interpenetrated into sodium alginate (SA) network to obtain self-enhanced injectable hydrogel. The optimum ratio of PVA/SA/BAG hydrogel was determined based on injectability, gelation time and chemical characterization. Results showed that the selected ratio had the shortest gelation time of 3.5min, and the hydrogel had a rough surface and good coagulation property. The hydrogel was capable of carrying 1kg of weight by mineralization for 14 d The compressive strength, compressive modulus, and fracture energy of the hydrogel reached 0.12MPa, 0.376MPa and 17.750kJ m−2, respectively. Meanwhile, the hydrogel had high moisture content and dissolution rate, and it was sensitive to temperature and ionic strength. Hydroxyapatite was generated on the hydrogel surface, and the hydrogel pores increased, and the pore size enlarged. The biocompatibility of PVA/SA/BAG hydrogel was analyzed using hemolysis and cytotoxicity assays. Results revealed its good biocompatibility with low hemolysis rate and no cytotoxicity to MC3T3-E1 cells. The hydrogel was also found to promote the differentiation of MC3T3-E1 cells with significantly increased in ALP activity and expression of relevant differentiation factors. In vitro mineralization assay showed an increase in calcium nodules and calcification area, indicating the ability of hydrogel to promote mineralization MC3T3-E1 cells. These findings indicated that PVA/SA/BAG hydrogel had potential uses in the field of irregular bone-defect repair due to its injectability, cytocompatibility, and tailorable functionality.

Serum Bile Acids Concentrations and Liver Enzyme Activities after Low-Dose Trilostane in Dogs with Hyperadrenocorticism.

Hyperadrenocorticism (HAC) often leads to vacuolar hepatopathy. The impact of trilostane treatment on serum total bile acids (SBAs) concentrations in dogs with HAC remains unknown. This study investigated SBAs concentrations in healthy dogs and those with HAC following trilostane therapy. Ten healthy dogs and fifteen dogs with HAC were prospectively enrolled. A biochemistry profile and pre- and post-prandial SBAs concentrations were determined in each dog. Dogs with HAC were reassessed at 1 and 3 months after the initiation of trilostane treatment. Dogs with HAC had significantly higher serum ALT, ALP, and GGT activities, and cholesterol, triglyceride, and pre-prandial SBAs concentrations compared to healthy dogs. After 3 months of trilostane treatment, polyuria/polydipsia and polyphagia were completely resolved in 42.8% and 35.7%, respectively. Significant improvements in serum ALT and ALP activities and cholesterol concentrations were observed within 1-3 months of trilostane treatment. However, pre- and post-prandial SBAs concentrations did not significantly decrease. These findings suggest that treatment with low-dose trilostane for 3 months appears to reduce serum liver enzyme activities, but not SBAs concentrations. Further investigation is warranted to explore the effects of low-dose trilostane treatment on SBAs concentrations for a longer duration or after achieving appropriate post-ACTH cortisol levels.

Hepato-Protective Activities of Ethanol Extracts of African Eggplant and Bitter Leaf in Alloxan Induced Diabetic Rats

Diabetes is a metabolic disorder, which affect all system of the body including the liver. This study focused on the effect of the ethanol extract of African eggplant and bitter leaf and their combination on the liver structure, antioxidant enzymes activities and biochemical parameters in the liver of alloxan induced diabetic rats. Twenty-five male wistar rats were grouped into normal control group A, diabetic control group B, 100 mg/kg African eggplant extract treated group C, 100 mg/kg bitter leaf extract treated group D and combination of 100 mg/kg of African eggplant and 100 mg/kg bitter leaf treated group E. The results study showed that group E rats showed significant reduction in the activity of AST, ALP and AST. In the antioxidant enzyme activity, the administered extracts increased the antioxidant enzyme activities and reduced lipid peroxidation, thereby protecting the cells from oxidative stress. The micro- histo-architecture of the liver showed improvement and regenerative changes across the treated groups. The study reveal that combination of the ethanol leaf extract of bitter leaf and African eggplant can manage liver malfunction caused by diabetes.

AuNP-Loaded Electrospinning Membrane Cooperated with CDs for Periodontal Tissue Engineering.

Guided bone regeneration (GBR) is commonly used to regenerate periodontal tissue. However, the bone inductivity and antibacterial properties of the GBR membranes currently in use are severely limited. This issue can be resolved by loading growth factors and antibiotics. Bioactive substitutes, such as Aunanoparticles (AuNPs) and carbon quantum dots (CDs), were proposed to prevent the denaturation of osteogenic growth factors and the induction of antibacterial drug resistance. Ornidazole was initially used as the raw material to prepare the CDs, followed by the incorporation of an optimal ratio of nanoparticles to produce the electrospun membrane doped with AuNPs and novel traceable antibacterial CDs. The morphology of the membrane was characterized. The adhesion, proliferation, and osteogenic differentiation of cells on the membrane were evaluated in vitro. The antimicrobial characteristics of the membrane were also investigated. The electrospun membrane was implanted into a rat skull defect model in vivo to investigate its osteogenic potential. The blending of nanomaterials did not affect the micro morphology of the fiber, resulting in enhanced mechanical properties. Membranes dopedwith AuNPs and CDs exhibited excellent biocompatibility, increased ALP activity, improved calcified nodules, and increased expression of osteogenic-associated proteins, in addition to pronounced antibacterial effects. The membrane also demonstrated excellent osteogenic characteristics in rat models. The synergistic effect of loaded AuNPs electrospun fiber membrane with CDs can promote periodontal bone regeneration and exert antibacterial activity.

Icariin attenuates the calcification of vascular smooth muscle cells through ERα - p38MAPK pathway.

To investigate the relationship between icariin and the osteoblastic differentiation of vascular smooth muscle cells (VSMCs) and the signal pathway involved. We applied a universally accepted calcification model of VSMCs induced by β glycerophosphate. Then the VSMCs calcification was observed by treatment with icariin and/or inhibitors of estrogen receptors (ERs) and p38-mitogen-activated protein kinase (MAPK) signaling. Icariin inhibited osteoblastic differentiation and mineralization of VSMCs due to decreased ALP activity and Runx2 expression. Further study demonstrated that icariin exerted this suppression effect through activating p38-MAPK but not extracellular-regulated kinase, JNK or Akt. An inhibitor of p38-MAPK partially reversed the inhibitory effects of icariin on osteoblastic differentiation. Interestingly, treatment of VSMCs with an ER antagonist ICI182780 and a selective ERα receptor antagonist PPT attenuated icariin-mediated inhibition effect of VSMCs calcification, associated with suppression of p38-MAPK phosphorylation. Icariin inhibited the osteoblastic differentiation of VSMCs, and that the inhibitory effects were mediated by p38-MAPK pathways through ERα.

Influence of METTL3 knockdown on PDLSC osteogenesis in E. coli LPS-induced inflammation.

This study aimed to investigate the effect of METTL3 knockdown on osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) in the weak inflammation microenvironments, as well as the underlying mechanisms. PDLSCs were stimulated by lipopolysaccharide from Escherichia coli (E. coli LPS), followed by quantification of METTL3. METTL3 expression was assessed using RT-qPCR and Western blot analysis in periodontitis. METTL3 knockdown PDLSCs were stimulated with or without E. coli LPS. The evaluation included proinflammatory cytokines, osteogenic markers, ALP activity, and mineralized nodules. Bioinformatics analysis and Western blot determined the association between METTL3 and the PI3K/Akt pathway. METTL3 was overexpressed in periodontitis. METTL3 knockdown in PDLSCs reduced proinflammatory cytokines, osteogenic markers, ALP activity, and mineralized nodules in both environments. Bioinformatics analysis suggested a link between METTL3 and the PI3K/Akt pathway. METTL3 knockdown inhibited PI3K/Akt signaling pathway activation. METTL3 knockdown might inhibit osteogenesis in PDLSCs through the inactivation of PI3K/Akt signaling pathway. Concomitant findings might shed novel light on the roles and potential mechanisms of METTL3 in the LPS-stimulated inflammatory microenvironments of PDLSCs.