Higher ALP Levels Research Articles

Hydrothermally crystallized Sr-containing bioactive glass film and its cytocompatibility

Abstract Sr-containing bioactive glass film with the Sr/(Ca+Sr) atomic ratio of 20% was sol-gel coated on titanium substrate, followed by hydrothermal treatment in a mixed phosphate solution (8 mM CaHPO 4 and 2 mM SrHPO 4 ) and other media at 120 or 140 °C. X-ray diffraction (XRD) analysis confirms amorphous nature of the gel powder calcined at 610 °C, but small peaks of Ca 1.8 Sr 0.2 SiO 4 or Ca 1.5 Sr 0.5 SiO 4 are also present. The absorption bands of Si-O-Si, PO 4 and OH groups and a weak absorption band of NO 3 groups appear in FT-IR spectrum of the calcined gel powder. Hydroxyapatite (HA) is detected for the hydrothermally treated sample by XRD and Raman analyses. Sr-containing HA nanocrystallites are formed on the film through a dissolution and precipitation mechanism in the hydrothermal treatment. The appearant bonding strength is 21±1 MPa for the both samples. In the test with MC3T3-E1 cells, the two coated samples exhibit larger viability, higher ALP levels and better cell morphology than the polished sample.

Calcium phosphates and silicon: exploring methods of incorporation

BackgroundBioinorganics have been explored as additives to ceramic bone graft substitutes with the aim to improve their performance in repair and regeneration of large bone defects. Silicon (Si), an essential trace element involved in the processes related to bone formation and remodeling, was shown not only to enhance osteoblasts proliferation but also to stimulate the differentiation of mesenchymal stem cells (MSCs) and preosteoblasts into the osteogenic lineage. In this study, the added value of Si to calcium phosphate (CaP) coatings was evaluated.MethodsTissue culture plastic well plates were coated with a thin CaP layer to which traces amounts of Si were added, either by adsorption or by incorporation through coprecipitation. The physicochemical and structural properties of the coatings were characterized and the dissolution behavior was evaluated. The adsorption/incorporation of Si was successfully achieved and incorporated ions were released from the CaP coatings. Human MSCs were cultured on the coatings to examine the effects of Si on cell proliferation and osteogenic differentiation. For the statistical analysis, a one-way ANOVA with Bonferroni post-hoc test was performed.ResultsThe results showed that human MSCs (hMSCs) responded to the presence of Si in the CaP coatings, in a dose-dependent manner. An increase in the expression of markers of osteogenic differentiation by human MSCs was observed as a result of the increase in Si concentration.ConclusionsThe incorporation/adsorption of Si into CaP coatings was successfully achieved and hMSCs responded with an increase in osteogenic genes expression with the increase of Si concentration. Furthermore, hMSCs cultured on CaP-I coatings expressed higher levels of ALP and OP, indicating that this may be the preferred method of incorporation of bioinorganics into CaPs.

Clinical-laboratory findings of bone metabolism in healthy premature and full-term neonates: preliminary results.

Premature infants are a major risk group for bone metabolic disorders. The purpose of this study is to clarify certain aspects of bone metabolism in healthy preterm and full-term neonates. Forty neonates (20 preterm and 20 full-term) were the material of the study. For each neonate demographic data (gender, gestational week) and anthropometric data (body weight) were recorded. Blood samples were collected and biochemical markers of bone metabolism (serum ALP, Ca, P, Mg) were immediately estimated. According to the results there is a statistically significant difference in average ALP of preterm neonates compared to full term neonates. Slightly higher values of Ca, P, Mg occurred in premature neonates while there was a statistically significant difference in the weeks of gestation and body weights between the two groups. It is typical in premature neonates the decrease in levels of ALP by the weeks of gestation and the stable levels of Ca. Gestational week seems to positively affect P and Mg levels in preterm neonates. Conclusively from our study's results arises that the week of gestation and not so much the body weight influence the alterations of bone biochemical biomarkers in healthy premature newborns. It seems that very premature neonates have high levels of serum ALP in decompensation of lower levels of Mg and P from all the newborns in this study. Therefore in very premature neonates, it is recommended to estimate serum ALP, Mg and P for assessment of bone turnover.

Evaluation of preoperative ultrasonographic and biochemical features of patients with aggressive parathyroid disease: is there a reliable predictive marker?

Parathyroid cancer (PC) represents < 1% of cases of PHPT. Tumors demonstrating atypical histopathologic features and don't fulfill criteria for carcinoma are classified as atypical adenomas (APA). The purpose of this study was to determine a biochemical or ultrasonographic feature that can predict aggressive disease requiring more extensive surgery and closer follow-up. Twenty eight patients operated for PHPT and diagnosed with atypical adenoma (23 patients) or carcinoma (5 patients) were enrolled in this study. The control group consisted of 102 patients operated between the same dates and diagnosed with classical PA. Classical adenomas, atypical adenomas, and carcinomas were compared according to their biochemical and ultrasonographic parameters. Serum Ca levels were significantly higher in the PC group compared with the APA and classical PA groups. Serum median PTH, Serum ALP and UCa was significantly higher in the APA and carcinoma groups compared to the classical PA group. ROC analysis was made to determine the best cut off values for predicting aggressive disease were 12.45 mg/dL, 265.05 pg/mL, 154.5 IU/l, 348.5 mg/day and 21.5 mm for Ca, PTH, ALP, UCa and the adenoma diameter, respectively. Multivariate analysis showed that serum Ca, ALP and isoechoic/cystic appearance were independent predictors for aggressive disease. Preoperatively high PTH, ALP, and UCa levels and large lesions with isoechoic or cystic appearances may be predictive of atypical adenoma or carcinoma in patients being evaluated for PHPT. In such cases, surgeons may prefer en bloc parathyroidectomy to minimally invasive surgery.

Avaliação da frequência de efeitos colaterais e distúrbios metabólicos em cães tratados com prednisone e deflazacorte

The aim of this study was to evaluate the frequency of adverse and metabolic effects caused by glucocorticoids used as antipruritic therapy in dogs. Two groups were studied. Group 1 consisted of six dogs treated with 0.5 mg/kg prednisone every 24 hours for seven days, subsequently on alternate days for one week, and then twice a week until 30 days of treatment were completed. Group 2 consisted of six dogs treated with 0.2 mg/kg deflazacort every 72 hours for 30 days. Clinical and laboratory changes were already evident 15 days after treatment onset in both groups. However, deflazacort promoted more metabolical alterations compared to prednisone with respect to hyperlipidemia, hyperglycemia, and high levels of ALT and ALP. These values improved after the interval between prednisone administrations was increased, which reinforces the importance of protocols with spaced dosage in chronic therapies.

Increased Serum Alkaline Phosphatase in a FeLV-Positive Cat

Background: Feline Leukemia Virus (FeLV) is an oncogenic virus that usually invades bone marrow causing non-regenerative anemia and thrombocytopenia. In FeLV positive cats, the myeloid leukemia is characterized by intense proliferation of blast cells in a bone marrow with myeloid precursor predominance and several malignancies degree. The leukocyte alkaline phosphatase is found secondary granules of mature neutrophils of humans, horses and cows but not in dogs and cats,which demonstrate the enzyme activity in myeloblast and promyeloblasts cells. In this case is describing a FeLV-positive case with leukemic disorder accompanied by extremely high levels of serum alkaline phosphatase.Case: A 8-year-old castrated male Domestic Short Hair cat with a 3-month history of convulsions, pica, vomits, lethargy, hypodipsia and anorexia was examined and showed mild dehydration, severe jaundice and hyperpnoea. Laboratory results evidenced severe normocytic hypochromic non-regenerative anemia, moderated leukocytosis with severe degenerativeneutrophil left shift, moderated lymphocytosis and severe thrombocytopenia. Serum biochemical profle showed mild hyperalbuminemia, moderated increment in alanine transaminase and severe increment in alkaline phosphatese. The cat waspositive for ELISA point of care FeLV antigen and negative for FIV antibodies. Urinalysis revealed moderated bilirubinuria and proteinuria. The bone marrow cytology showed high cellularity, increased of M:E ratio, rare megakaryocyte lineage, erythroid hypoplasia, myeloid hyperplasia with blast excess and dysgranulopoiesis.Discussion: FeLV is a virus that usually causes cytopenias and in this case this was well evidenced through a severe non regenerative anemia and thrombocytopenia. Generally, anemia is a non-neoplastic complication of FeLV mainly related to suppressive effects hematopoietic and stromal cells of the bone marrow. Nevertheless, in this case, the anemia also maybe attributable to the observed acute myeloid leukemia. Neutrophilia is an uncommon fnding in FeLV-infected cats, even more with the severe degenerative left shift as in the present case. This fnding could be associated to a myeloid hyperplasia. Some biochemical alterations, as hyperalbuminemia and increment in values of serum ALT could be explained by clinical observations like dehydration and some degree of hepatic enlargement, respectively. However, the excessive value forserum alkaline phosphatase activity is an unpublished fnding in cats. In humans high levels of ALP has been associated with neutrophilia and it is quite likely that a great amount of degenerative neutrophils may contribute for high leakage of ALP than in other circumstances of neutrophilia. This is the frst report of a FeLV-positive cat exhibiting extremely high serum values of the enzyme ALP, possibly attributable to acute myeloid leukemia.Keywords: neutrophilia, cytopenia, non-regenerative anemia, acute leukemia.

Properties of Flavonoids Isolated from the Bark of Eysenhardtia polystachya and Their Effect on Oxidative Stress in Streptozotocin-Induced Diabetes Mellitus in Mice.

Six new flavonoids 2′,4′-dihydroxychalcone-6′-O-β-d-glucopyranoside (1), α,3,2′,4′-tetrahydroxy-4-methoxy-dihydrochalcone-3′-C-β-glucopyranosy-6′-O-β-d-glucopyranoside (2), 7-hydroxy-5,8′-dimethoxy-6′α-l-rhamnopyranosyl-8-(3-phenyl-trans-acryloyl)-1-benzopyran-2-one (3), 6′7-dihydroxy-5,8-dimethoxy-8(3-phenyl-trans-acryloyl)-1-benzopyran-2-one (4), 9-hydroxy-3,8-dimethoxy-4-prenylpterocarpan (5), and α,4,4′-trihydroxydihydrochalcone-2′-O-β-d-glucopyranoside (6) were isolated from bark of Eysenhardtia polystachya. Antidiabetic activity of compounds 1–5 in terms of their cellular antioxidant and free radical scavenging and also in streptozotocin- (STZ-) induced diabetic mice was evaluated on liver transaminases, lipid peroxidation, total bilirubin, total protein, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (CSH-Px), and glutathione reductase (GSH). Results indicated that 1–5 scavenged 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (∙OH), nitric oxide radicals (NO∙), superoxide anion radical (O2 ∙−), radical cation (ABTS∙+), and hydrogen peroxide (H2O2) radical, and protection against H2O2 induced BSA damage was also observed. Furthermore, 1–5 showed ability to decrease the oxidative stress in H9c2 cell. Diabetic mice present high levels of lipid peroxide, total protein, SGPT, SGOT, ALP, and TB. However, treatment of STZ-induced diabetes in mice with 1–5 reduced levels of these enzymes leading to protector effect of liver. In addition, with treatment with 1–5, increases in radical scavenging enzymes of CSH-Px, SOD, GSH, and CAT have also been observed in diabetic mice. The antioxidant properties of compounds 1–5 are a promising strategy for ameliorating therapeutic effects by avoiding disorders in the normal redox reactions in healthy cells which consequently could alleviate complications of diabetes.

High serum ALP level is associated with increased risk of denosumab-related hypocalcemia in patients with bone metastases from solid tumors.

Metastatic bone disease is one of the most common complications of advanced cancers. Pathological fractures, spinal cord compression, and radiotherapy or surgery to the bone are collectively called skeletal-related events (SREs), which cause severe pain, increase hospitalization rates, and impair the quality of life (QOL) of patients with bone metastases. The receptor activator of nuclear factor-kB ligand (RANKL)/RANK pathway is critical in the progression of bone metastases. Previous studies have demonstrated that an anti-RANKL antibody (denosumab) was superior to zoledronic acid in prolonging time to first SRE in patients with bone metastases from prostate and breast cancers. However, severe hypocalcemic events occur more frequently after treatment with denosumab compared with zoledronic acid. In this study, 368 administrations of denosumab in 219 patients with metastatic bone disease from solid tumors were analyzed to clarify the risk factors for developing hypocalcemia. The results showed that grade 2/3 hypocalcemia was observed in 10.4% of the total number of denosumab administrations. Patients with higher baseline serum ALP, higher performance status (PS), or gastric cancer were at higher risk for developing hypocalcemia. The cut-off value for ALP to predict denosumab-related hypocalcemia was 587 U/L with a sensitivity of 0.77 and a specificity of 0.81. Close monitoring of serum calcium, especially after the first treatment with denosumab, is strongly recommended in these patients.

RE1-Silencing Transcription Factor (Rest) is a Novel Regulator of Osteoblast Differentiation.

RE1-silencing transcription factor (Rest) has been identified as a master negative regulator of neuronal differentiation. Nothing is known about Rest function in bone cells. In this study, we examined the Rest expression levels and role during osteoblast differentiation. We found that Rest is abundantly expressed in bone marrow stromal cells, calvarial osteoblasts, and MC3T3-E1 osteoblasts. Treatment of primary osteoblasts with ascorbic acid (AA) down regulated Rest mRNA expression at an early stage, but not in later stages of differentiation. Consistent with treatment of primary cultures, AA treatment of MC3T3-E1 cells significantly reduced Rest protein expression at day 3 and at day 8 after initiation of osteoblast differentiation. Treatment of bone marrow stromal cells with BMP-2 and dexamethasone, but not IGF-I for 3 days greatly decreased Rest mRNA expression. To test the function of Rest during osteoblast differentiation, Rest expression was knocked down in MC3T3-E1 cell subclones segregated on the basis of ALP activity (differentiation status) using lentivirus expressing shRNA against Rest. An 80% knockdown of Rest expression decreased Osterix (Osx) expression by 52-57% and as a result, increased both basal and AA induced ALP expression and activity in the subclone that expresses low basal level of ALP (undifferentiated). By contrast, a 98% knockdown of Rest expression in cells that express high basal levels of ALP (differentiated cells) caused a significant reduction in Osx expression, basal and AA induced ALP expression and activity. These data suggest that Rest regulates early osteoblast differentiation via modulating Rest expression that is independent of Osx expression.

Zebrafish: A possible tool to evaluate bioactive ions

Zebrafish is a well-established model organism with a skeletal structure that highly resembles mammalian bone. Yet its use in the research field of biomaterials has been limited. One area that could benefit from this model system is the evaluation of ionic dissolution products from different materials. As a proof of concept we have evaluated the effect of silicate ions on the zebrafish larvae and compared it to a well-known osteblastic cell line, MC3T3-E1 subclone 14. We have shown that sodium metasilicate (125μM and 625μM) induces more mineralisation in a dose-dependent manner in zebrafish larvae, 9days post fertilisation as compared to the non-treated group. Moreover the same trends were seen when adding sodium metasilicate to MC3T3-E1 cultures, with more mineralisation and higher ALP levels with higher doses of silicate (25, 125 and 625μM). These results indicate the feasibility of zebrafish larvae for ionic dissolution studies. The zebrafish model is superior to isolated cell cultures in the aspect that it includes the whole bone remodelling system, with osteoblasts, osteoclasts and osteocytes. Zebrafish could thus provide a powerful in vivo tool and be a bridge between cell culture systems and mammalian models.

Age and sex specific variation in hematological and serum biochemical parameters of Beluga (Huso huso Linnaeus, 1758)

In the present study, the age- and sex-specific changes of various haematological and blood serum biochemical blood parameters of Beluga ( Huso huso ) were investigated. Blood samples were collected from 4-, 6-, 7-, and 8-year-old beluga (n = 7 for each sex and age). The specimens were fed at a rate of 0.5-3% body weight per day. AST and LDH levels in 7- and 8-year-old fish of both sexes were significantly higher ( P <0.05) than those in 4- and 6-year-old individuals. The mean ALT were significantly different ( P <0.05) in both sexes of 4-, 6-, and 7-year-old sturgeon. However, the 6-, 7-, and 8-year-old female sturgeon had higher ALP levels ( P <0.05). Additionally, mean RBC, PCV, and Hb values were significantly higher ( P <0.05) in 7- and 8-year-old females and males than the others. Two-tailed Pearson’s correlation between the biochemical and haematological parameters obtained for beluga sturgeon indicated significant positive correlations between AST and ALP, AST and LDH, ALP and LDH, RBC and Hb, RBC and PCV, Hb and PCV, MCH and MCHC, and MCV and MCH. However, significant negative correlations were found between RBC and MCV and MCH. These results suggest that the blood parameters of beluga are influenced by age- and sex-specific factors.

Normal Bone Turnover in Transient Hyperphosphatasemia

Transient hyperphosphatasemia of infancy and early childhood (THI) is characterized by a temporary isolated elevation of serum alkaline phosphatase activity (ALP), predominantly its bone or liver isoform, in either sick or healthy children under 5 years of age. Return to normal ALP levels usually occurs within four months. Spontaneous rise of ALP might concern the physician, especially when treating seriously ill children. However, THI is considered a benign biochemical disorder with no clinical consequences. Some existing reports support the hypothesis that THI is a result of increased bone turnover. We present evidence of normal bone turnover in two children with THI. In a one-year-old girl and a boy of the same age, high ALP levels (31 and 109 μkat/L, respectively) were accidentally detected. The children had no signs of metabolic bone disease or of liver disease. The high ALP levels returned to normal in two months, thus fulfilling the diagnosis of THI. In both patients, serum parathyroid hormone and bone turnover markers, serum CrossLaps, and serum osteocalcin were neither elevated, nor did these markers follow the ALP dynamics, thus reflecting normal bone turnover in THI. Children with THI should be spared from extensive investigations and unnecessary vitamin D treatment. Conflict of interest:None declared.

P2-49 Subclinical diastolic dysfunction is associated with biomarkers of hepatic function: results from the stop HF study

IntroductionThis analysis set to investigate the relationship between novel biomarkers of cardiovascular morbidity and mortality with diastolic dysfunction in a primary care cohort at heightened cardiovascular risk.MethodsThis is a cross-sectional...

Bone turnover markers for osteoporotic status assessment? A systematic review of their diagnosis value at baseline in osteoporosis

Objective Osteoporosis diagnosis is based on bone mineral density (BMD) but bone remodeling is also a crucial issue. It can be assessed by bone turnover markers (BTMs). Their interest for the positive and etiological diagnosis of osteoporosis at baseline, and their predictive value for past asymptomatic vertebral fractures, were evaluated by a systematic review of the literature. Methods Medline database was searched to identify all published reports analyzing BTMs and BMD or fractures. We conducted meta-analyses on BTMs levels according to osteoporotic status using random effects models. Results Moderate and negative correlations were found, mainly in postmenopausal women, between BTMs and BMD, especially with bone alkaline phosphatase (bone ALP), osteocalcin, serum C-terminal and urine N-terminal crosslinking telopeptides of type I collagen (sCTX and uNTX). Bone ALP and sCTX levels are higher in osteoporotic patients compared to controls. High levels of bone ALP in primary hyperparathyroidism and low levels of osteocalcin in endogenous hypercorticism are the most relevant data reported in endocrine diseases associated with osteoporosis. High levels of BTMs, especially osteocalcin, bone ALP or sCTX, may be associated with prevalent vertebral fractures. Conclusion The diagnosis value of BTMs at baseline in osteoporosis is very low. The interest of BTMs for the etiological diagnostic of secondary osteoporosis has not been demonstrated. Data are lacking to address the interest of BTMs assessment to screen for vertebral fractures in asymptomatic patients with high risk factors of fractures.

IN VITRO STUDY OF THE EFFECT OF SOME PREPARED OXADIAZOLE DERIVATIVES ON ALP, AST AND ALT ACTIVITIES IN SERA OF HEPATOCELLULAR CANCER PATIENTS AND HEALTHY CONTROL

The effect of some prepared oxadiazole compounds 5-[4-(p-methoxy phenyl) azo phenyl]-2thiol-1, 3, 4-Oxadiazole [ ], 5-[4-(methoxy phenyl) azo phenyl]-2-marcapto-alky-1, 3, 4-oxadiazole [ -V]( where alkyl group is ethyl, propyl, butyl and pentyl respectively) were tested on the activity of ALP, AST, and ALT in sera of 30 patients with hepatocellular cancer and 30 healthy individuals as control group. The results revealed significantly higher levels of ALP, AST, and ALT activity in patients group compared to control group. Different concentrations (10 -4 , 10 -5 , and 10 -6 M) of oxadiazole derivatives showed different percentage of inhibitions, but did not showed proportional with the concentration of derivatives. The elevated values of ALP, AST, and ALT activity in sera of patients were returned to about normal values in most cases after addition of oxadiazole compounds to sera of patients in vitro. Keyword: Oxadiazole compounds, Anticancer, Enzymatic Biomarker.

Anti-hyperlipidemic and Cardioprotective Effects of Ocimum sanctum L. Fixed Oil in Rats Fed a High Fat Diet

Ocimum sanctum (OS) has a lipid-lowering action in both normal and diabetic animals. Because OS leaves are rich in oil, the present study was conducted to explain the anti-hyperlipidemic and organ-protective effect of OS fixed oil in rats fed with a high fat (HF) diet. OS fixed oil was extracted by hexane and the fatty acids composition identified by GC-MS. Four groups of male Wistar rats included a normal control group, a high fat fed-diet (HF) group, a HF group treated with OS fixed oil, and a HF group treated with a reference drug simvastatin. The results show that OS fixed oil contains five kinds of fatty acids, of which alpha-linolenic acid was the major fatty acid. OS fixed oil depressed high serum levels of total cholesterol, triglyceride, LDL-C, and AI, whereas no significant effect on HDL-C was observed. OS fixed oil also suppressed high levels of liver cholesterol and triglyceride with no significant effect on both lipids in feces. In addition, OS fixed oil normalized the high serum levels of LDH and CK-MB but no significant effect on high serum levels of ALT, AST, and ALP was obtained. We conclude that treatment with OS fixed oil during the last three weeks of HF diet feeding decreased the high serum lipid profile and expressed antiartherogenic and cardioprotective actions against hyperlipidemia. The anti-hyperlipidemic action of OS fixed oil was mainly resulted from the suppression of liver lipid synthesis. Linolenic acid and linoleic acid contained in OS fixed oil were possibly responsible for both lipid-lowering and cardiac protective action against hyperlipidemia.

The significance of antimitochondrial IgA and IgG in the diagnosis of primary biliary cirrhosis

To evaluate the sensitivity and the specificity of Anti-M2-3E ELISA for the detection of IgG- and IgA-specific isotypes of antimitochondrial antibody (AMA), and to investigate the significance of antimitochondrial IgA and IgG in the diagnosis of primary biliary cirrhosis (PBC). Sera were collected from 107 PBC patients, 87 disease controls and 26 healthy controls, and the antimitochondrial antibodies (IgG and IgA) were detected using indirect immunofluorescence (IFL), Anti-PDC ELISA and Anti-M2-3E ELISA. The AMA IgG positive rate in PBC patients was 90.6% detected by Anti-M2-3E ELISA, which is significantly higher than that (81.3%) detected by IFL(t = 4.32, P < 0.05) and that (72.9%) detected by Anti- PDC ELISA (t = 6.03, P < 0.05). The AMA IgA was positive in 59 of the 107 PBC patients, and 99 of the 107 patients were positive for AMA IgG or/and IgA. 9 of the 20 IFL-negative patients were positive for AMA IgG as indicated by Anti-M2-3E ELISA, 11 of the 20 IFL-negative patients were positive for AMA IgG or/and IgA as indicated Anti-M2-3E ELISA. Compared to patients negative for IgG AMA, patients positive for IgG AMA had more severe histopathology and higher levels of ALP, IgG, and IgM. The IgG and IgA Anti- M2-3E ELISAs are more sensitive for the AMA detection than IFN and the Anti-PDC ELISA. The presence of AMA IgG is the characteristics of severe PBC.

Mesenchymal stem cell proliferation and differentiation on an injectable calcium phosphate – Chitosan composite scaffold

Calcium phosphate cement (CPC) can be molded or injected to form a scaffold in situ, has excellent osteoconductivity, and can be resorbed and replaced by new bone. However, its low strength limits CPC to non-stress-bearing repairs. Chitosan could be used to reinforce CPC, but mesenchymal stem cell (MSC) interactions with CPC-chitosan scaffold have not been examined. The objective of this study was to investigate MSC proliferation and osteogenic differentiation on high-strength CPC-chitosan scaffold. MSCs were harvested from rat bone marrow. At CPC powder/liquid (P/L) mass ratio of 2, flexural strength (mean ± sd; n = 5) was (10.0 ± 1.1) MPa for CPC-chitosan, higher than (3.7 ± 0.6) MPa for CPC ( p < 0.05). At P/L of 3, strength was (15.7 ± 1.7) MPa for CPC-chitosan, higher than (10.2 ± 1.8) MPa for CPC ( p < 0.05). Percentage of live MSCs attaching to scaffolds increased from 85% at 1 day to 99% at 14 days. There were (180 ± 37) cells/mm 2 on scaffold at 1 day; cells proliferated to (1808 ± 317) cells/mm 2 at 14 days. SEM showed MSCs with healthy spreading and anchored on nano-apatite crystals via cytoplasmic processes. Alkaline phosphatase activity (ALP) was (557 ± 171) (pNPP mM/min)/(μg DNA) for MSCs on CPC-chitosan, higher than (159 ± 47) on CPC ( p < 0.05). Both were higher than (35 ± 32) of baseline ALP for undifferentiated MSCs on tissue-culture plastic ( p < 0.05). In summary, CPC-chitosan scaffold had higher strength than CPC. MSC proliferation on CPC-chitosan matched that of the FDA-approved CPC control. MSCs on the scaffolds differentiated down the osteogenic lineage and expressed high levels of bone marker ALP. Hence, the stronger CPC-chitosan scaffold may be useful for stem cell-based bone regeneration in moderate load-bearing maxillofacial and orthopedic applications.

Osteogenic potential of the human bone morphogenetic protein 2 gene activated nanobone putty

Nanobone putty is an injectable and bioresorbable bone substitute. The neutral-pH putty resembles hard bone tissue, does not contain polymers or plasticizers, and is self-setting and nearly isothermic, properties which are helpful for the adhesion, proliferation, and function of bone cells. The aim of this study was to investigate the osteogenic potential of human bone morphogenetic protein 2 (hBMP2) gene activated nanobone putty in inducing ectopic bone formation, and the effects of the hBMP2 gene activated nanobone putty on repairing bone defects. Twenty four Kunming mice were randomly divided into two groups. The nanobone putty + hBMP2 plasmid was injected into the right thigh muscle pouches of the mice (experiment side). The nanobone putty + blank plasmid or nanobone putty was injected into the left thigh muscle pouches of the group 1 (control side 1) or group 2 (control side 2), respectively. The effects of ectopic bone formation were evaluated by radiography, histology, and molecular biology analysis at 2 and 4 weeks after operation. Bilateral 15 mm radial defects were made in forty-eight rabbits. These rabbits were randomly divided into three groups: Group A, nanobone putty + hBMP2 plasmid; Group B, putty + blank plasmid; Group C, nanobone putty only. Six rabbits with left radial defects served as blank controls. The effect of bone repairing was evaluated by radiography, histology, molecular biology, and biomechanical analysis at 4, 8, and 12 weeks after operation. The tissue from the experimental side of the mice expressed hBMP2. Obvious cartilage and island-distributed immature bone formation in implants of the experiment side were observed at 2 weeks after operation, and massive mature bone observed at 4 weeks. No bone formation was observed in the control side of the mice. The ALP activity in the experiment side of the mice was higher than that in the control side. The tissue of Group A rabbits expressed hBMP2 protein and higher ALP level. The new bone formation rate and antibending strength of group A was significantly higher than those of group B and C. The defects in blank control were not healed. The hBMP2 gene activated nanobone putty exhibited osteoinductive ability, and had a better bone defect repair capability than that of nanobone putty only.

Mucin-producing Bile Duct Carcinoma Arising from Primary Sclerosing Cholangitis: A Case Report

A 60-year-old woman with primary sclerosing cholangitis (PSC) and high levels of ALP, gamma-GTP, and DUPAN-2 was admitted to our institution for examination. The patient did not have ulcerative colitis or pancreatic intraductal papillary mucinous neoplasm. Imaging studies revealed atypical dilation of bile ducts in the left lobe of the liver. Repeated cytologic examinations of the bile showed atypical cells consistent with adenocarcinoma. The patient underwent extended resection of the left lobe of the liver and was found to have intraductal papillary carcinoma with associated mucin-producing bile duct carcinoma. This carcinoma fills dilated bile duct lumens with mucin. This tumor differs morphologically from typical cholangiocarcinoma, which is usually seen in the late stages of PSC. Just one case of mucin-producing bile duct carcinoma arising from PSC has been reported worldwide. The patient has had no signs of recurrence after 27 months. Patients with mucin-producing bile duct carcinoma, as in the case of its pancreatic counterpart, may have a better prognosis and a higher survival rate than patients with typical cholangiocarcinomas.