Small Diameter, High Surface Energy Carbon Nanofiber Formulations that Selectively Increase Osteoblast Function

2001 ◽  
Vol 711 ◽  
Author(s):  
Rachel L. Price ◽  
Kathy L. Elias ◽  
Karen M. Haberstroh ◽  
Thomas J. Webster

ABSTRACTThe objective of the present in vitro study was to investigate the potential of carbon nanofibers, which have nanometer dimensions similar to hydroxyapatite crystals in physiological bone, for orthopedic applications. Studies of alkaline phosphatase activity and calcium deposition by osteoblasts (the bone-synthesizing cells) were performed on both nanophase (less than 100 nm) and conventional (greater than 100 nm) diameter carbon nanofibers. Results provided the first evidence of a strong correlation between decreased carbon fiber diameter and both increased alkaline phosphatase activity and increased calcium deposition by osteoblasts at early time points (specifically, 7 days), but not at later time points (specifically, 14 and 21 days). Results of early calcium deposition by osteoblasts on carbon nanofibers are promising and consistent with the desired rapid formation of natural bone at the implant interface.

Author(s):  
Kari Hanson ◽  
Carly Isder ◽  
Kristen Shogren ◽  
Anthony L. Mikula ◽  
Lichun Lu ◽  
...  

OBJECTIVE The use of intrawound vancomycin powder in spine surgery has been shown to decrease the rate of surgical site infections; however, the optimal dose is unknown. High-dose vancomycin inhibits osteoblast proliferation in vitro and may decrease the rate of solid arthrodesis. Bone marrow–derived mesenchymal stem cells (BMSCs) are multipotent cells that are a source of osteogenesis in spine fusions. The purpose of this study was to determine the effects of vancomycin on rat BMSC viability and differentiation in vitro. METHODS BMSCs were isolated from the femurs of immature female rats, cultured, and then split into two equal groups; half were treated to stimulate osteoblastic differentiation and half were not. Osteogenesis was stimulated by the addition of 50 µg/mL l-ascorbic acid, 10 mM β-glycerol phosphate, and 0.1 µM dexamethasone. Vancomycin was added to cell culture medium at concentrations of 0, 0.04, 0.4, or 4 mg/mL. Early differentiation was determined by alkaline phosphatase activity (4 days posttreatment) and late differentiation by alizarin red staining for mineralization (9 days posttreatment). Cell viability was determined at both the early and late time points by measurement of formazan colorimetric product. RESULTS Viability within the first 4 days decreased with high-dose vancomycin treatment, with cells receiving 4 mg/mL vancomycin having 40%–60% viability compared to the control. A gradual decrease in alizarin red staining and nodule formation was observed with increasing vancomycin doses. In the presence of the osteogenic factors, vancomycin did not have deleterious effects on alkaline phosphatase activity, whereas a trend toward reduced activity was seen in the absence of osteogenic factors when compared to osteogenically treated cells. CONCLUSIONS Vancomycin reduced BMSC viability and impaired late osteogenic differentiation with high-dose treatment. Therefore, the inhibitory effects of high-dose vancomycin on spinal fusion may result from both reduced BMSC viability and some impairment of osteogenic differentiation.


RSC Advances ◽  
2015 ◽  
Vol 5 (99) ◽  
pp. 81378-81387 ◽  
Author(s):  
Ting Ma ◽  
Xi-Yuan Ge ◽  
Sheng-Nan Jia ◽  
Xi Jiang ◽  
Yu Zhang ◽  
...  

The effect of alkali-treated titanium surfaces on inflammation-related gene expression of macrophages and alkaline phosphatase activity of osteoblast-like cells.


1992 ◽  
Vol 262 (3) ◽  
pp. F442-F448 ◽  
Author(s):  
N. S. Krieger ◽  
N. E. Sessler ◽  
D. A. Bushinsky

Metabolic acidosis induces net calcium flux (JCa) from cultured neonatal mouse calvariae through physicochemical and cell-mediated mechanisms. To determine the role of osteoblasts in acid-induced JCa, collagen synthesis and alkaline phosphatase activity were assessed in calvariae incubated in reduced pH and bicarbonate medium, a model of metabolic acidosis (Met), and compared with controls (Ctl). Collagen synthesis fell from 30.5 +/- 1.1 in Ctl to 25.1 +/- 0.4% with Met, and alkaline phosphatase decreased from 403 +/- 25 in Ctl to 298 +/- 21 nmol Pi.min-1.mg protein-1 with Met. During acidosis JCa was correlated inversely with percent collagen synthesis (r = -0.743, n = 11, P = 0.009) and with alkaline phosphatase activity (r = -0.453, n = 22, P = 0.034). To determine the role of osteoclasts in acid-induced JCa, osteoclastic beta-glucuronidase activity was determined in Ctl and Met in the absence or presence of the osteoclastic inhibitor calcitonin (CT, 3 x 10(-9) M). Met increased beta-glucuronidase (5.9 +/- 0.2) compared with Ctl (4.6 +/- 0.3 micrograms phenolphthalein released.bone-1.h-1), whereas CT inhibited beta-glucuronidase in both Ctl and Met (3.1 +/- 0.2 and 3.5 +/- 0.3, respectively). During acidosis JCa was correlated directly with beta-glucuronidase activity (r = 0.683, n = 42, P less than 0.001). Thus the cell-mediated component of JCa during acidosis in vitro appears to result from a combination of inhibited osteoblastic and stimulated osteoclastic activity.


Blood ◽  
1965 ◽  
Vol 25 (3) ◽  
pp. 356-369 ◽  
Author(s):  
FRED ROSNER ◽  
STANLEY L. LEE

Abstract Leukocyte alkaline phosphatase activity has been noted to be different in men and women. The mean leukocyte alkaline phosphatase activity for 74 normal men, aged 19 to 60 years, was 23 mg. of phosphorus per 1010 polvmorphonuclear leukocytes per hour. The corresponding mean value for 75 normal young women, age 19-48 years, was 35 (p < .001). No significant differences between boys and girls occurred until the time of puberty. After the menopause, the values for women approached the values for men. Women treated with androgens had lower leukocyte alkaline phosphatase activity than did control women. These results suggest that androgenic hormones inhibit this enzyme, and that other, as yet undefined endocrine influences, also affect its level of activity. In vitro tests with various concentrations of androgens and estrogens failed to provide conclusive evidence of direct effect on leukocytes although some degree of direct inhibition by androgens was suggested. Studies using saponin to effect enzyme release from leukocyte granules did not demonstrate whether the differences between men and women are differences of enzyme release or of content of leukocyte alkaline phosphatase.


2007 ◽  
Vol 2 (11) ◽  
pp. 1934578X0700201 ◽  
Author(s):  
Chau Van Minh ◽  
Nguyen Xuan Cuong ◽  
Tran Anh Tuan ◽  
Eun Mi Choi ◽  
Young Ho Kim ◽  
...  

Chemical investigation of the Vietnamese soft coral Sarcophyton mililatensis resulted in the isolation of a new 9,11-secosterol, sarcomilasterol (1), along with two known polyhydroxylated sterols, sarcoaldesterol B (2) and ergosta-1β,3β,5α,6β-tetraol (3). Their structures were established on the basis of NMR spectroscopic and MS experiments. Compound 1 (3 μM) significantly increased the growth of MC3T3-E1 cells and caused a significant elevation of alkaline phosphatase activity and nodules mineralization compared to those of the control (P<0.05). These results suggest that compound 1 has a direct stimulatory effect on bone formation in vitro and may contribute to the prevention of osteoporosis.


Cerâmica ◽  
2012 ◽  
Vol 58 (348) ◽  
pp. 481-488 ◽  
Author(s):  
F. A. Macedo ◽  
E. H. M. Nunes ◽  
W. L. Vasconcelos ◽  
R. A. Santos ◽  
R. D. Sinisterra ◽  
...  

Highly porous three-dimensional biodegradable scaffolds was obtained from beta-tricalcium phosphate-hydroxyapatite bioceramic (BCP), PCL, and Angiotensin-(1-7). We used the solvent casting and particulate leaching methods (SC/PL). The processed scaffolds were characterized by X-ray microtomography (µ-CT). Biocompatibility tests in vitro were performed during three and seven days using MTT and Alkaline Phosphatase Activity (APA) assays. Both the MTT activity and APA were evaluated using a one-way ANOVA test. The µ-CT results showed that the increase of the PCL:BCP weight ratio leads to structures with lower pore sizes. The pore interconnectivity of the processed scaffolds was evaluated in terms of the fragmentation index (FI). We observed that the obtained composites present poorly connected structures, with close values of FI. However, as the polymer phase is almost transparent to the X-rays, it was not taken into consideration in the µ-CT tests. The MTT activity assay revealed that scaffolds obtained with and without Angiotensin-(1-7) present mild and moderate cytotoxic effects, respectively. The APA assay showed that the rat osteoblasts, when in contact for three days with the PCL composites, presented an APA similar to that observed for the control cells. Nevertheless, for an incubation time of seven days we observed a remarkable decrease in the alkaline phosphatase activity. In conclusion, using the solvent casting and salt leaching method we obtained 3D porous that are composites of PCL, BC and Ang-(1-7), which have suitable shapes for the bone defects, a high porosity and interconnect pores. Furthermore, the viability in vitro showed that the scaffolds have potential for drug delivery system and could be used in future in vivo tests.


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