Bone-related Markers Research Articles

Osteoking inhibits apoptosis of BMSCs in osteoporotic rats via PI3K/AKT signaling pathway.

This study aims to investigate whether Osteoking inhibits apoptosis of BMSCs in osteoporotic rats via the PI3K/AKT signaling pathway and to conduct a detailed exploration of this mechanism. The goal is to provide a theoretical basis for the clinical application of Osteoking in osteoporosis treatment. A rat model of osteoporosis was established through bilateral ovariectomy (OVX), followed by treatment with Osteoking. After ten weeks of therapy, BMD was evaluated. The biomechanics of the left tibia were measured, the left femur was sequenced, and the right tibia was stained using histomorphometric and Masson's staining methods. Peripheral serum was collected to measure bone-related markers, including E2, PINP, and CTX. RNA-Seq results were verified using the remaining bone samples. Comparative analysis demonstrated the efficacy of Osteoking in treating osteoporosis and provided preliminary insights into the underlying mechanisms. Primary BMSCs were cultured using bone marrow apposition. CCK8 assays were conducted to screen the intervention conditions of Osteoking and LY294002. Various concentrations of Osteoking-containing serum and LY294002 were tested separately to determine the optimal intervention concentration for drug delivery. The impact of Osteoking on lipid formation was also evaluated. Following treatment of BMSCs from OVX rats with Sham serum, OVX serum, OVX+LY294002 serum, and Osteoking+LY294002 serum, the expression of PI3K/AKT/mTOR, osteogenesis-related regulatory factors, and apoptosis-related regulatory factors was assessed. Flow cytometry was employed to evaluate apoptosis in BMSCs. Osteoking significantly improved whole-body BMD and bone biomechanical indices in OVX rats. It also significantly elevated the serum levels of E2 and PINP while reducing the level of CTX, which significantly improved bone microstructure and promoted new bone formation. RNA-seq analysis indicated that the therapeutic mechanism involved the PI3K/AKT signaling pathway. Osteoking increased the expression of RUNX2 and decreased the expression of PPAR-γ, a marker of lipogenesis, in OVX rats. Extraction of BMSCs for subsequent studies revealed a significant reduction in proliferation and osteogenic differentiation, along with an increase in lipogenic differentiation, in the OVX group. Osteoking treatment inhibited the expression of PPAR-γ and increased the expression of RUNX2 in BMSCs. Additionally, Osteoking reversed the LY294002-mediated inhibition of PI3K/AKT/mTOR signaling pathway activation, increased the expression of the apoptosis-protecting protein Bcl2, and decreased the expression of apoptosis-associated proteins Caspase3 and Bax. Osteoking markedly improved bone microstructure, biomechanics, and bone density in OVX rats. Osteoking-containing serum reversed the imbalance in lineage differentiation in OVX rats, characterized by reduced osteogenic differentiation and increased lipid differentiation of BMSCs. Furthermore, Osteoking-containing serum significantly increased BMSC proliferation and prevented apoptosis in OVX rats through the PI3K/AKT signaling pathway.

Comparative analysis of bone health and glycemic control in postmenopausal women: insights from clinical data

Abstract Background Menopause leads to significant physiological and biochemical alterations that impact different aspects of health, such as bone mineral metabolism and glycemic control. It is imperative to comprehend these alterations in order to identify potential health hazards and develop preventative measures. This comparative cross-sectional study aimed to evaluate bone-associated markers and glycemic control indices in postmenopausal women (PMP). Methods A sample of 100 women aged 30 years or older was randomly selected and categorized into three groups: PMP (n = 60), premenopausal (PRM) (n = 20), and women in the reproductive age group (RWA) (n = 20). Venous blood samples were collected, and fasting blood glucose (FBG), calcium, phosphorus, uric acid, and alkaline phosphatase (ALP) levels were analyzed using standard laboratory techniques. The insulin and estrogen levels were evaluated using enzyme-linked immunosorbent assay (ELISA), and Homeostasis model assessment of insulin resistance (HOMA-IR), Quantitative insulin sensitivity check index (QUICKI), and Fasting insulin resistance index (FIRI) were calculated. A significance level of p < 0.05 was used in the statistical analysis conducted using SPSS version 25.0. Results The PMP and PRM groups showed significantly greater mean values of FBG, insulin, and HOMA-IR when compared to RWA (p < 0.05), while these groups had lower QUICKI levels. Similar trends were noted for bone-associated markers, with significant differences among the groups (p < 0.05). The correlation results showed that QUICKI had positive correlations with both ALP and phosphorus, while FBG and FIRI had negative connections with both. Conclusion Postmenopausal women demonstrated higher levels of glycemic control indices and bone-related markers compared to premenopausal and reproductive-age women, indicating potential risks for osteoporosis and obesity.

Pilot study of the effect of surgical menopause on bone mineral density and quality in patients with gynecological malignancies.

To investigate the effects of surgical menopause on bone mineral density and bone quality because bilateral salpingo-oophorectomy for the treatment of gynecological malignancies is common even in premenopausal patients. This study is prospective one of bone mineral density and quality measurements after surgery for perimenopausal gynecologic malignancies. In 50 women who underwent surgical menopause for a diagnosis of gynecological malignancies, bone mineral density (BMD), blood levels of tartrate-resistant acid phosphatase 5b (TRACP-5b) and bone-specific alkaline phosphatase (BAP) as bone metabolism markers, and urinary pentosidine level as bone quality marker were measured before surgery and at multiple points up to 24 months after surgery. In a group of 22 patients who did not undergo hormone replacement therapy (HRT) (HRT- group), BMD of the lumbar spine and total hip continued to decrease significantly from 6 months postoperatively. Percentages of changes in BMD progressively increased over time after surgery. TRACP-5b and urinary pentosidine levels significantly increased 6 months postoperatively compared with preoperative levels. Comparisons between 10 patients who underwent HRT (HRT+ group) and the HRT- group revealed significant reductions in the percentage of change in lumbar spine BMD only and TRACP-5b and urinary pentosidine levels 12 months postoperatively in the former group. In this pilot study, we showed that BMD and bone-related markers are altered in patients with surgical menopause. It also suggested that HRT may reduce these influences on bone metabolism.

Hormone Replacement Therapy Relieves Periodontitis by Inhibiting Alveolar Bone Loss and Inflammation.

Hormone replacement therapy (HRT) is a commonly used strategy for treating menopausal symptoms, while its relation with periodontitis remains unclear. This study aimed to explore the potential effects of HRT on periodontitis, mainly in aspects of bone loss and inflammation. The alveolar bone height (ABH), alveolar bone thickness (ABT), and bone mineral density (BMD) were measured in menopausal women with periodontitis who had received HRT or had not received HRT by cone beam computed tomography. Based on a rat model of periodontitis, the alveolar bone loss was evaluated by micro-computed tomography and bone-related biochemical markers. The expression/levels of inflammatory markers were measured to reflect periodontal inflammation. Although the differences were not all significant in each premolars/molars, the mesial/distal ABH and buccal/lingual ABT were lower, and the mesial/distal BMD was higher in patients in the HRT group than those in the control group. In a rat model of periodontitis, the alveolar bone loss was relieved by HRT. Additionally, HRT significantly weakened the elevation of inflammatory markers, including TNF-α, IL-1β, and IL-6 in periodontitis rats. HRT contributes to the remission of periodontitis by inhibiting alveolar bone loss and inflammation.

The association of vitamin D insufficiency with the prevalence of obesity in children: implications for serum calcium levels, alkaline phosphatase activity, and bone maturation.

Vitamin D deficiency has been identified as a potential risk factor for various adverse health outcomes. However, its specific role in metabolic regulation and skeletal development in school-aged children is not fully understood. This study aimed to explore the correlation between vitamin D deficiency and childhood obesity rates, and its impact on serum calcium, alkaline phosphatase, and bone age in children. The study analyzed clinical data from 159 school-aged children who underwent medical examinations. Participants were divided into the 25-hydroxyvitamin D3 (25(OH)D3) deficiency group and the 25(OH)D3 normal group based on their serum levels. We compared body mass index (BMI), total cholesterol (TC), triglycerides (TG), Ca, ALP, bone age, fasting blood glucose (FBG), and hemoglobin A1c (HbA1c) between the two groups. Logistic regression and Spearman correlation analyses were performed to further investigate relationships between 25(OH)D3 levels and metabolic and bone-related markers. This study showed that the 25(OH)D3 deficiency cohort exhibited significantly higher BMI, TC, TG, and ALP levels, with lower Ca levels and delayed bone age compared to the normal group. Logistic regression analysis identified Ca, ALP, and bone age as significant predictors of 25(OH)D3 deficiency. Subgroup analysis showed that in the 25(OH)D3 deficient group, children with higher BMI had elevated TC, ALP levels, and delayed bone age, while Ca levels were lower. Correlation analysis confirmed the predictive value of these markers for 25(OH)D3 deficiency. Our findings demonstrate that 25(OH)D3 deficiency is strongly associated with obesity in school-aged children and may negatively affect normal skeletal development. Regular monitoring of 25(OH)D3 levels in school-aged children is essential for ensuring proper growth and development, especially in those at risk for obesity.

Relationship Between Serum 25-Hydroxyvitamin D and Bone Mineral Density, Fracture Risk, and Bone Metabolism in Adults With Osteoporosis/Fractures

ObjectiveTo evaluate the association of serum 25-hydroxyvitamin D (25(OH) D) levels with bone mineral density (BMD), fracture risk, and bone metabolism. MethodsThis multicenter cross-sectional study recruited menopausal females and males greater than or equal to 50 year old with osteoporosis/fractures between September 2016 and September 2021. Assessment included clinical data, 25(OH)D, intact parathyroid hormone (iPTH), procollagen type 1 amino-terminal propeptide (P1NP), carboxy-terminal collagen crosslinks (CTX), lateral thoracolumbar spine x-rays, and BMD. ResultsA total of 3003 individuals were stratified by 25(OH) D levels: 720 individuals (24%) <20 ng/mL, 1338 individuals (44.5%) 20 to 29 ng/mL, and 945 individuals (31.5%) ≥30 ng/mL. In unadjusted and multivariable models, BMD T-score, except spine, was significantly and positively associated with 25(OH)D levels. 25(OH) D levels were inversely associated with Fracture Risk Assessment Tool scores. Patients with 25(OH)D <20 ng/mL had significantly higher iPTH and bone turnover markers (P1NP and CTX) than patients with 25(OH)D ≧20 ng/mL in all models. When analyzing bone-related markers and BMD, total hip and femoral neck BMD T-scores were positively correlated with 25(OH)D concentrations and BMI but negatively correlated with iPTH, P1NP, CTX, and age. In multivariate models with all bone-related markers, only 25(OH)D levels were significantly associated with total hip and femoral neck BMD. ConclusionVitamin D deficiency is significantly associated with decreased total hip and femoral neck BMD and increased fracture risk as assessed by Fracture Risk Assessment Tool. In those with osteoporosis/fractures, vitamin D is implicated in the causal relationship between bone remodeling and BMD. Assessing vitamin D status is imperative for those at risk for osteoporosis/fractures.

Branched Channels in Porous β-Tricalcium Phosphate Scaffold Promote Vascularization.

Rapid and efficient vascularization is still considerably challenging for a porous β-tricalcium phosphate (β-TCP) scaffold to achieve. To overcome this challenge, branched channels were created in the porous β-TCP scaffold by using 3D printing and a template-casting method to facilitate the instant flow of blood supply. Human bone mesenchymal stem cells (hBMSCs) and human umbilical vein endothelial cells (HUVECs) were seeded in the channeled porous scaffolds and characterized through a double-stranded DNA (dsDNA) assay, alkaline phosphatase (ALP) assay, and cell migration. Channeled porous β-TCP scaffolds were then implanted in the subcutaneous pockets of mice. Histological staining and immunohistochemical staining on vascularization and bone-related markers were carried out on the embedded paraffin sections. Results from in vitro experiments showed that branched channels significantly promoted HUVECs' infiltration, migration, proliferation, and angiogenesis, and also promoted the proliferation and osteogenesis differentiation of hBMSCs. In vivo implantation results showed that, in the early stage after implantation, cells significantly migrated into branched channeled scaffolds. More matured blood vessels formed in the branched channeled scaffolds compared to that in nonchanneled and straight channeled scaffolds. Beside promoting vascularization, the branched channels also stimulated the infiltration of bone-related cells into the scaffolds. These results suggested that the geometric design of branched channels in the porous β-TCP scaffold promoted rapid vascularization and potentially stimulated bone cells recruitment.

Conducting biointerface of spider-net-like chitosan-adorned polyurethane/SPIONs@SrO2–fMWCNTs for bone tissue engineering and antibacterial efficacy

In pursuit of enhancing bone cell proliferation, this study delves into the fabrication of porous scaffolds through the integration of nanomaterials. Specifically, we present the development of highly conductive chitosan (CS) nanonets on fibro-porous polyurethane (PU) bio-membranes. These nanofibers comprise functionalized multiwall carbon nanotubes (fMWCNTs), well-dispersed superparamagnetic iron oxide (SPIONs), and strontium oxide (SrO2) nanoparticles. The resulting porous scaffold exhibits remarkable interfacial biocompatibility, antibacterial properties, and load-bearing capability. Through meticulous in vitro investigations, the CS-PU/SPIONs/SrO2-fMWCNTs nanofibrous scaffolds have demonstrated a propensity to promote bone cell regeneration. Notably, the integration of these nanomaterials has been found to upregulate crucial bone-related markers, including ALP, ARS, COL-I, RUNX2, and SPP–I. The evaluation of these markers, conducted through quantitative real-time polymerase chain reaction (qRT-PCR) and immunocytochemistry, substantiates the improved cell survival and enhanced osteogenic differentiation facilitated by the integrated nanomaterials. This comprehensive analysis underscores the efficacy of CS-PU/SPIONs/SrO2-fMWCNTs bioscaffolds in promoting MC3T3-E1 cell regeneration within, thereby holding promise for advancements in bone tissue engineering and regenerative medicine.

Electrochemical technique to develop surface-controlled polyaniline nano-tulips (PANINTs) on PCL-reinforced chitosan functionalized (CS-f-Fe2O3) scaffolds for stimulating osteoporotic bone regeneration

Bone defects pose significant challenges in orthopedic surgery, often leading to suboptimal outcomes and complications. Addressing these challenges, we employed a three-electrode electrochemical system to fabricate surface-controlled polyaniline nano-tulips (PANINTs) decorated polycaprolactone (PCL) reinforced chitosan functionalized iron oxide nanoparticles (CS-f-Fe2O3) scaffolds. These structures were designed to emulate the natural extracellular matrix (ECM) and promote enhanced osseointegration by establishing a continuous interface between host bone and graft, thereby improving both biological processes and mechanical stability. In vitro experiments demonstrated that PANINTs-PCL/CS-f-Fe2O3 substrates significantly promoted the proliferation, differentiation, and spontaneous outgrowth and extension of MC3T3-E1 cell activity. The nanomaterials exhibited increased cell viability and osteogenic differentiation, as evidenced by elevated expression of bone-related markers such as ALP, ARS, COL-I, RUNX2, and SPP-I, as determined by qRT-PCR. Our findings underscore the regenerative potential of in situ cell culture systems for bone defects, emphasizing the targeted stimulation of essential cell subpopulations to facilitate rapid bone tissue regeneration.

The Phenotype of Mesenchymal Stromal Cell and Articular Chondrocyte Cocultures on Highly Porous Bilayer Poly-L-Lactic Acid Scaffolds Produced by Thermally Induced Phase Separation and Supplemented with Hydroxyapatite.

Bilayer scaffolds could provide a suitable topology for osteochondral defect repair mimicking cartilage and subchondral bone architecture. Hence, they could facilitate the chondro- and osteogenic lineage commitment of multipotent mesenchymal stromal cells (MSCs) with hydroxyapatite, the major inorganic component of bone, stimulating osteogenesis. Highly porous poly-L-lactic acid (PLLA) scaffolds with two layers of different pore sizes (100 and 250 µm) and hydroxyapatite (HA) supplementation were established by thermally induced phase separation (TIPS) to study growth and osteogenesis of human (h) MSCs. The topology of the scaffold prepared via TIPS was characterized using scanning electron microscopy (SEM), a microCT scan, pycnometry and gravimetric analysis. HMSCs and porcine articular chondrocytes (pACs) were seeded on the PLLA scaffolds without/with 5% HA for 1 and 7 days, and the cell attachment, survival, morphology, proliferation and gene expression of cartilage- and bone-related markers as well as sulfated glycosaminoglycan (sGAG) synthesis were monitored. All scaffold variants were cytocompatible, and hMSCs survived for the whole culture period. Cross-sections revealed living cells that also colonized inner scaffold areas, producing an extracellular matrix (ECM) containing sGAGs. The gene expression of cartilage and bone markers could be detected. HA represents a cytocompatible supplement in PLLA composite scaffolds intended for osteochondral defects.

Physiologic Effects of Isolated or Synthetic Dietary Fiber in Children: A Scoping Review

BackgroundFiber is an integral part of a healthy diet. Studies have shown that the fiber intake in children is below adequate amounts, leading to adverse health outcomes. ObjectivesThis study aimed to perform a scoping review to assess the available evidence for the impact of isolated and synthetic dietary fiber on children’s health outcomes. MethodsA systematic literature search was conducted in Ovid Medline, Ovid Global Health, Embase, and Cochrane Library via Wiley to identify randomized controlled trials (RCTs) in healthy children aged 1–18 y at baseline who consumed added, isolated, or synthetic dietary fiber. The outcomes of interest were categorized based on the Food and Drug Administration’s guidance for industry on nondigestible carbohydrates and the Vahouny Fiber Symposium criteria, which included reduced fasting blood, glucose, total and/or LDL cholesterol concentrations, attenuation of postprandial glycemia/insulinemia, increased fecal bulk/laxation, reduced transit time, weight loss/reduction in adiposity, reduced energy intake from food consumption, increased satiety, bone health/enhanced mineral absorption, and blood pressure. We also cataloged additional reported outcomes. ResultsOf 3837 randomized controlled parallel or crossover trials screened at the abstract level, 160 were eligible for full-text review, and 32 included for data extraction. This scoping review presents analysis of data from 32 RCTs in children who were healthy, overweight/obese or had mild hypercholesterolemia. Inulin-type fructans (41%) and psyllium (22%) were the most frequently administered fiber types, with weight/adiposity, markers of lipid metabolism (41%), and bone-related markers (38%) being the most frequently reported health outcomes. Only a few RCTs have investigated the effects of laxation (9%), and none specifically studied the impact of fiber on reducing postprandial glycemia/insulinemia. ConclusionsThis scoping review demonstrates sufficient evidence for conducting systematic reviews and meta-analyses for several outcomes. Evidence gaps remain on the impact of isolated fibers on outcomes such as laxation, colonic transit time, and postprandial glycemia/insulinemia in children.

Laser-Modified Ti Surface Improves Paracrine Osteogenesis by Modulating the Expression of DKK1 in Osteoblasts

Titanium surface modifications are widely used to modulate cellular behavior by recognition of topographical cues. However, how those modifications affect the expression of mediators that will influence neighboring cells is still elusive. This study aimed to evaluate the effects of conditioned media from osteoblasts cultured on laser-modified titanium surfaces on the differentiation of bone marrow cells in a paracrine manner and to analyze the expression of Wnt pathway inhibitors. Mice calvarial osteoblasts were seeded on polished (P) and Yb:YAG laser-irradiated (L) Ti surfaces. Osteoblast culture media were collected and filtered on alternate days to stimulate mice BMCs. Resazurin assay was performed every other day for 20 days to check BMC viability and proliferation. After 7 and 14 days of BMCs maintained with osteoblasts P and L-conditioned media, alkaline phosphatase activity, Alizarin Red staining, and RT-qPCR were performed. ELISA of conditioned media was conducted to investigate the expression of Wnt inhibitors Dickkopf-1 (DKK1) and Sclerostin (SOST). BMCs showed increased mineralized nodule formation and alkaline phosphatase activity. The L-conditioned media enhanced the BMC mRNA expression of bone-related markers Bglap, Alpl, and Sp7. L-conditioned media decreased the expression of DKK1 compared with P-conditioned media. The contact of osteoblasts with Yb:YAG laser-modified Ti surfaces induces the regulation of the expression of mediators that affect the osteoblastic differentiation of neighboring cells. DKK1 is among these regulated mediators.

Ethnic determinants of skeletal health in female patients with fragility fracture in a culturally diverse population

BackgroundLow bone density leads to fragility fracture, with significant impact on morbidity and mortality. While ethnic differences in bone density have been observed in healthy subjects, this has not yet been explored in fragility fracture patients. AimsTo assess if ethnicity is associated with bone mineral density and serum markers of bone health in female patients who experience fragility fractures. Methods219 female patients presenting with at least one fragility fracture at a major tertiary hospital in Western Sydney Australia were studied. Western Sydney is a region with great cultural diversity, comprising people from over 170 ethnicities. Within this cohort, the three largest broad ethnic groups were Caucasians (62.1 %), Asians (22.8 %), and Middle Eastern patients (15.1 %). Location and nature of the presenting fracture and other relevant past medical history were obtained. Bone mineral density, measured by dual-energy X-ray absorptiometry, and bone-related serum markers were compared between ethnicities. Covariates (age, height, weight, diabetes, smoking, and at-risk drinking) were adjusted in multiple linear regression model. ResultsAlthough Asian ethnicity was associated with lower bone mineral density at the lumbar spine in fragility fracture patients, this association was no longer significant after adjustment for weight. Ethnicity (Asian or Middle Eastern) was not a determinant of bone mineral density at any other skeletal site. Caucasians had lower estimated glomerular filtration rate compared to Asian and Middle Eastern subjects. Serum parathyroid hormone concentrations were significantly lower in Asians compared to other ethnicities. ConclusionAsian ethnicity and Middle Eastern ethnicity were not major determinants of bone mineral density at the lumbar spine, femoral neck, or total hip.

Influence of partially exposed nonabsorbable membrane for alveolar ridge preservation: A randomized controlled trial.

This randomized controlled trial evaluated the impact of a partially exposed non-absorbable membrane (dPTFE) in Alveolar Ridge Preservation (ARP) procedures on clinical, tomographic, immunoenzymatic, implant-related, and patient-centered outcomes. Patients with a hopeless maxillary single-rooted tooth demanding rehabilitation with implants were included. Patients were randomized into two groups: dPTFE (n=22)-tooth extraction followed by ARP using a partially exposed dPTFE membrane; USH (n=22)-unassisted socket healing. Clinical and tomographic analyses were performed at baseline and after 3 months. After 3 months, patients received one dental implant. Implant stability quotient was obtained following implant placement. Bone-related markers were analyzed in bone biopsies using an immunoenzymatic assay. Greater gain in Keratinized Mucosa Width (KMW) was observed in the dPTFE (1.33 ± 0.98 mm) compared to USH (0.59± 0.98 mm) (Mann-Whitney test, Z=2,28, p < 0.05). USH showed a reduction of pain/discomfort, edema, and interference with daily life from the seventh day (Friedman/Wilcoxon test, maxT=7.48, 8.00, and 5.92, respectively, p < 0.05). dPTFE presents a reduction of edema and interference with daily life from the 7th day and pain/discomfort from the 14th day (Friedman/Wilcoxon test, maxT=5.40, 5.26, and 4.78, respectively, p < 0.05). The dPTFE group presented higher pain/discomfort in the 35 and 42 days and higher edema from 7 to 42 days postoperatively than USH group (Mann-Whitney test, p < 0.05). No differences between groups were observed in the tomographic measures, immunoenzymatic analysis, and implant stability (p > 0.05). dPTFE was superior to USH by increasing KMW gain. However, dPTFE without bone graft presented similar bone loss compared to USH. This clinical trial was not registered prior to participant recruitment and randomization (NCT04329351).

Sika deer velvet antler protein extract modulater bone metabolism and the structure of gut microbiota in ovariectomized mice.

Osteoporosis is a systemic osteopathy characterized by bone metabolism disorders that become more serious with age increases in postmenopausal women. Recent studies have found that antler protein is the main bioactive component of cervus pantotrichum, and it has a positive regulatory effect on bone metabolism and can improve estrogen level. This study aimed to investigate the effect of velvet antler extract (VAE) on the prevention of osteoporosis and the modulation of gut microbiota in ovariectomized (OVX) mice. OVX mice treated with 12 weeks of VAE exhibited higher levels of serum BGP, Ca2+, CT, and HyP (p < .05). Micro-CT scans showed that VAE significantly elevated bone volume fraction (BV/TV), trabecular bone number (Tb.N), trabecular bone thickness (Tb.Th), trabecular bone connection density (Conn.D), decreased trabecular separation (Tb.Sp), and structural modality index (SMI) than untreated OVX mice. The right tibial retinaculum in the VAE group was clearer, with a clearer reticular structure, smaller gaps, a tighter distribution, and a more orderly arrangement. The gut microbiota of the cecal contents was analyzed by 16 s rDNA amplicon sequencing. The data indicated that VAE modulated the species, numbers, and diversity of the gut microbiota in OVX mice. Ovariectomy caused dysbiosis of the intestinal microbiota by increasing the ratio of Firmicutes to Bacteroidetes in mice, but the ratio decreased after treatment with VAE. These results suggest that VAE has a therapeutic effect on OVX mice via modulate bone-related biochemical markers in serum and structure of gut microbiota.

Impact of serum vitamin D level on selected bone-related markers in obese- type 2 diabetes patients

Background: Type 2 diabetes mellitus (T2DM) is accompanied by an increased risk for skeletal fractures. The causes are probably a mix of factors, including poor glycemic control, a higher risk of falling due to hypoglycemia, osteopenia, bone quality deterioration, and drug side effects related to bone fragility. Undercarboxylated osteocalcin (ucOC) is a protein generated by osteoblasts that affects insulin secretion and sensitivity. Methods: A total number of 47 obese (BMI ˃ 30) patients with confirmed type 2 diabetes were selected depending on the American Diabetes Association ADA criteria. The control participants were similar to the patients in age and gender, with a total number of 43 obese (BMI ˃ 30) and healthy subjects chosen from the general population. The selected subjects were grouped according to their serum vitamin D levels. Their blood specimen was used for assaying ucOC, parathyroid hormone (PTH), and vitamin D3 levels by specific ELISA kits, and to estimate calcium (Ca2+) levels and inorganic phosphate (PO43−) via enzymatic colorimetric methods. Results: Serum parathyroid hormone and inorganic phosphate median (IQR) values were markedly increased in patients with T2DM when compared to healthy controls, whereas serum calcium and ucOC levels were lowered significantly in diabetic patients when compared to healthy controls. This was irrespective of serum vitamin D levels. Conclusions: Elevated serum levels of PTH and PO43− values in obese type 2 diabetic patients compared to obese non-diabetic controls were accompanied by a significant decrease in ucOC and Ca2+ levels, irrespective of serum vitamin D levels. Hence, serum vitamin D3 levels had no significant impact on levels of ucOC, PTH, Ca2+, and PO43− in obese patients with type 2 diabetes.

Impact of a modified implant macrogeometry on biomechanical parameters and bone-related markers in rats.

This study investigated the impact of a modified implant macrogeometry on peri-implant healing and its effect on bone-related molecules in rats. Eighteen rats received one implant in each tibia: the control group received implants with conventional macrogeometry and the test group received implants with modified macrogeometry. After 30 days, the implants were removed for biomechanical analysis and the bone tissue around them was collected for quantifying gene expression of OPN, Runx2, β-catenin, BMP-2, Dkk1, and RANKL/OPG. Calcein and tetracycline fluorescent markers were used for analyzing newly formed bone at undecalcified sections of the tibial implants. These fluorescent markers showed continuous bone formation at cortical bone width and sparse new bone formed along the medullary implant surface in both groups. However, higher counter-torque values and upregulation of OPN expression were achieved by test implants when compared to controls. The modified macrogeometry of implants optimized peri-implant healing, favoring the modulation of OPN expression in the osseous tissue around the implants.

Does resveratrol favor peri-implant bone repair in rats with ovariectomy-induced osteoporosis? Gene expression, counter-torque and micro-CT analysis

This study investigated the influence of resveratrol on peri-implant repair and its effects on bone-related markers in ovariectomy-induced osteoporosis in rats. Animals were divided into: OVX+PLAC (n = 10): ovariectomized animals treated with placebo; OVX+RESV (n = 10): OVX treated with resveratrol; OVX+PLAC+ZOL (n = 10): OVX treated with PLAC and zoledronate; OVX+RESV+ZOL (n = 10): OVX treated with RESV and ZOL; and SHOVX+PLAC (n = 10): sham ovariectomy treated with PLAC. RESV and PLAC were administrated after ovariectomy and ZOL after six weeks after OVX, until the end of experiment. One implant was inserted in each tibiae of animals 18 weeks after ovariectomy. After 4 weeks, one implant was removed for counter-torque, and peri-implant tissue was collected for mRNA quantification of several osteogenic markers by PCR. The other tibia was submitted to micro-computed tomography analysis. Reduced counter-torque values, bone-implant contact (BIC) and bone volume fraction (BV/TV), and higher bone porosity (BP) were detected in OVX+PLAC group when compared to SHOVX+PLAC (p < 0.05). OVX+RESV rats presented lower BIC, BV/TV, and trabecular number (Tb.N), and augmented BP and trabecular spacing (Tb.Sp) when compared to SHOVX+PLAC (p < 0.05). Higher Tb.N and connectivity density (Conn.Dn) and reduced Tb.Sp were observed in OVX rats treated with ZOL, independently of RESV, when compared to OVX+PLAC and OVX+RESV groups (p < 0.05), whereas the combination ZOL+RESV promoted lower BP when compared to OVT+PLAC and OVX+RESV (p < 0.05). Gene expression was not influenced by RESV (p > 0.05), whereas ZOL promoted up-regulation of BMP-2 (p<0.05). RESV did not improve peri-implant bone repair in rats with ovariectomy-induced osteoporosis.

Repairing a critical cranial defect using WISP1-pretreated chondrocyte scaffolds

In cranial flat bone fractures, spontaneous bone repair will occur only when the fracture ends are in close contact. However, in cases wherein bone discontinuity is extensive, surgical interventions are often required. To this end, autologous bone is harvested and surgically integrated into the site of fracture. Here we propose to use cartilage, as an alternative autologous source, to promote cranial fracture repair. The advantage of this approach is the potential reduction in donor site morbidity, likely due to the avascular and aneural nature of cartilage. As a first step we attempted to induce cartilage mineralization in vitro, using micromass primary chondrocyte cultures, incubated with BMP2 and/or WISP1, which were examined histologically following a 3-week culture period. Next, chondrocyte seeded collagen scaffolds were evaluated in vitro for expression profiles and ALP activity. Finally, chondrocyte-seeded collagen scaffolds were implanted in a Lewis rats 8 mm critical calvaria defect model, which was imaged via live CT for 12 weeks until sacrifice. End points were analyzed for microCT, histology, and serum levels of bone related markers. Micromass cultures exhibited an osseous inducing trend following WISP1 administration, which was maintained in chondrocyte seeded scaffolds. Accordingly, in vivo analysis was carried out to assess the impact of WISP1-pretreated chondrocytes (WCS) versus untreated chondrocytes (UCS) in calvaria defect model and compared to untreated control comprised of a defect-associated blood clot (BC) or empty collagen scaffold (CS) implant. Live CT and microCT exhibited higher mineralization volumes in critical defect implanted with UCS, with some structural improvements in WCS. Histological analysis exhibited higher anabolic bone formation in WCS and trabecular bone was detected in WCS and UCS groups. Chondrocytes implanted into critical cranial defect expedite the formation of native-like osseous tissue, especially after WISP1 priming in culture. Ultimately, these data support the use of autologous chondrocytes to repair critical maxillofacial defects.

Micro/nanometer-sized porous structure of zinc phosphate incorporated Ti(HPO4)2 hydrate bioceramic induces osteogenic gene expression and enhances osteoporotic bone regeneration

In this work, we describe the synthesis of an ideal bioceramic to which cells can easily respond that has geometry and composition that are similar to those of bone-like apatite. The successful preparation of bone-inspired titanium hydrate phosphate with an optimized weight percentage (∼1.1 wt%) of zinc in (Ti(HPO4)2/Zn3(PO4)2·nH2O) bioceramic could represent a potential regenerative framework for bone defect repair and bone void filler that would be useful for various clinical approaches. We have for the first time demonstrated the detailed physicochemical and biological characterizations of the bioceramic required to achieve a biocompatible and porous architecture that is similar to hydroxyapatite. In vitro studies have shown that the phosphate-rich titanium and zinc nanocomposite promote bone regeneration after stimulating MC3T3 − E1 and hBM-MSCs cells activity. The metallic ions and phosphate had the effects of enhancing the cell viability and osteogenic differentiation of the cells, as confirmed by the expression of bone-related markers that included ALP, Col1a1, RUNX2, OPN/Spp1, and OCN through qRT-PCR, Western blotting, and immunocytochemistry. Moreover, the bioceramic has been shown to significantly improves new bone growth in critical size calvarial defect in rat model. The findings of the present work are the first that show the bone regeneration effect of bioceramic, which could be crucial for subsidies of Ca–P-based artificial bone implant materials. Our results provide insight into the Ti(HPO4)2/Zn3(PO4)2(0.1)·nH2O bioceramic in osteogenesis which can help to establish a therapeutic strategy in bone tissue regenerative medicine.