Olga García-Martínez

Effects of Indomethacin, Nimesulide, and Diclofenac on Human MG-63 Osteosarcoma Cell Line

Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most widely prescribed drugs worldwide and serve as treatment of some degenerative inflammatory joint diseases. The aim of the present study was to investigate the influence of different concentrations of three NSAIDs on cell proliferation, differentiation, antigenic profile, and cell cycle in the human MG-63 osteosarcoma cell line, incubated for 24 hr. All NSAIDs had an inhibiting effect on osteoblastic proliferation. Treatments for 24 hr had small but significant effects on the antigenic profile. No treatment altered osteocalcin synthesis. Indomethacin and nimesulide treatments arrested the cell cycle at G0/G1. These results suggest that indomethacin, nimesulide, and diclofenac appear to have no effects on osteocalcin synthesis and a slight effect on the antigenic profile. They may delay bone regeneration due to their inhibiting effect on osteoblast growth. Therefore, these drugs should only be used in situations that do not require rapid bone healing.

Effect of Platelet-Rich Plasma on Growth and Antigenic Profile of Human Osteoblasts and Its Clinical Impact

PURPOSEnIn recent years, there has been widespread clinical use of platelet-rich plasma (PRP) to facilitate the regeneration of different tissues. However, few data are available on the effect of PRP on parameters other than cell growth. The aim of the present study was to evaluate the effect of PRP on the cell cycle, antigenic profile, and proliferation of primary cultured human osteoblasts.nnnMATERIALS AND METHODSnThe cells in the present study were derived from human bone sections obtained from healthy volunteers during third molar surgery. PRP was prepared from human venous blood and used to culture the cell line obtained from the same patient. Flow cytometry was used to study the cell cycle, antigenic profile, and proliferation.nnnRESULTSnThe treatment of osteoblasts with PRP modified the expression of CD54, CD80, CD86, and HLA-DR antigens. PRP treatment increased cell proliferation in the short term, but the cell proliferation capacity diminished in the long term, perhaps owing to cell exhaustion. No change in the cell cycle profile was observed in the PRP-cultured cells.nnnCONCLUSIONSnThese results suggest that PRP treatment accelerates bone neoformation with no cell cycle changes that might carry a risk of malignant transformation.

Effect of acetaminophen (paracetamol) on human osteosarcoma cell line MG63

AbstractAim:To examine the effects of acetaminophen (paracetamol), a nonsteroidal anti-inflammatory drug (NSAID), on different cellular and functional parameters of the human osteosarcoma cell line MG63.Methods:Flow cytometry was used to study proliferation, antigenic profile, and phagocytic activity, and radioimmunoassay was used to determine osteocalcin synthesis as a cell differentiation marker.Results:Short-term treatment with therapeutic doses of paracetamol(5 or 25 μmol/L) reduced cell proliferation, osteocalcin synthesis, and phagocyte activity, and increased the expression of antigens involved in antigen presentation to T lymphocytes (CD80, CD86, HLA-DR).Conclusion:These findings suggest that paracetamol activates the osteoblast, inducing its immunogenic action to the detriment of its bone formation capacity.

Phenolic Compounds in Extra Virgin Olive Oil Stimulate Human Osteoblastic Cell Proliferation

In this study, we aimed to clarify the effects of phenolic compounds and extracts from different extra virgin olive oil (EVOO) varieties obtained from fruits of different ripening stages on osteoblast cells (MG-63) proliferation. Cell proliferation was increased by hydroxytyrosol, luteolin, apigenin, p-coumaric, caffeic, and ferulic acids by approximately 11–16%, as compared with controls that were treated with one vehicle alone, while (+)-pinoresinol, oleuropein, sinapic, vanillic acid and derivative (vanillin) did not affect cell proliferation. All phenolic extracts stimulated MG-63 cell growth, and they induced higher cell proliferation rates than individual compounds. The most effective EVOO phenolic extracts were those obtained from the Picual variety, as they significantly increased cell proliferation by 18–22%. Conversely, Arbequina phenolic extracts increased cell proliferation by 9–13%. A decline in osteoblast proliferation was observed in oils obtained from olive fruits collected at the end of the harvest period, as their total phenolic content decreases at this late stage. Further research on the signaling pathways of olive oil phenolic compounds involved in the processes and their metabolism should be carried out to develop new interventions and adjuvant therapies using EVOO for bone health (i.e.osteoporosis) in adulthood and the elderly.

Effects on Growth of Human Osteoblast-Like Cells of Three Nonsteroidal Anti-Inflammatory Drugs Metamizole, Dexketoprofen, and Ketorolac

Some nonsteroidal anti-inflammatory drugs (NSAIDs) have adverse effects on bone tissue. The objective of this study was to determine the effect of different doses of dexketoprofen, ketorolac, and metamizole on growth of the osteoblast MG63 cell line. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide spectrophotometry results showed that MG63 cell growth was significantly inhibited after 24 hr of culture with doses of 10, 20, 100, or 1,000 µM of each NSAID and with doses of 0.1, 1, or 5 µM of dexketoprofen and ketorolac but not metamizole. Cell-cycle studies revealed that dexketoprofen and ketorolac treatments significantly arrested the cell cycle in phase G0/G1, increasing the percentage of cells in this phase. Apoptosis/necrosis studies showed significant changes versus control cells, with an increased percentage of cells in apoptosis after treatment with 10, 100, or 1,000 µM of metamizole and after treatment with 1, 10, 100, or 1,000 µM of dexketoprofen or ketorolac. In conclusion, treatment of osteoblast-like cells with high doses of the NSAIDs tested increased not only the percentage of cells in apoptosis but also the percentage of necrotic cells.

Wettability and osteoblastic cell adhesion on ultrapolished commercially pure titanium surfaces: the role of the oxidation and pollution states

The oxidation state of the surfaces of titanium-based biomaterials strongly depends on their previous history. This factor affects the titanium wettability and it probably conditions the success of the implanted biomaterials. However, the separate role of the pollution and oxidation states of metallic titanium surfaces remains still controversial. To elucidate this, it is required to standardize the initial surface state of titanium in terms of roughness and surface chemistry, and then, to monitor its wettability after the corresponding treatment. In this work, we studied finely polished surfaces of commercially pure titanium (cpTi) which were subjected to cleaning surface treatments. X-Photoelectron spectroscopy was used to characterize the surface chemistry and the oxide film thickness. The contact angle hysteresis in underwater conditions was measured with the growing/shrinking captive bubble method, which allowed for mimicking the real conditions of implantable devices. The water wettability of smooth cpTi surfaces was stabilized with weak thermal oxidation (230u2009°C, 30u2009min). The osteoblastic cell response of the stabilized and non-stabilized cpTi surfaces was analyzed. Although the oxidation and pollution states were also stabilized and normalized, no correlation was observed between the stable response in wettability of titanium and its cell adhesion.

Human fibroblast-like cultures in the presence of platelet-rich plasma as a single growth factor source: clinical implications.

OBJECTIVE: The purpose of this study was to compare the proliferation, morphology, and antigenic expression of human fibroblast–like cells between primary cultures treated with platelet-rich plasma (PRP) or fetal bovine serum (FBS) as the growth factor source. DESIGN: Cells from human gingival tissue samples obtained from healthy volunteers during oral surgery were studied. Isolated cells were cultured in media supplemented with 10% PRP or FBS. Platelet-rich plasma was prepared from the venous blood of each patient. The authors studied short- and long-term cell cultures in the presence of PRP or FBS as the sole growth factor source in order to determine (a) cell growth rate, by MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay; (b) cell morphology, by electronic microscopy; and (c) antigenic expression, by flow cytometry and confocal microscopy. RESULTS: In short-term cultures, the cell growth rate was higher with PRP versus FBS treatment. No differences in morphology or expression of vimentin, fibronectin, or &agr;-actin antigens were observed between PRP and FBS cultures. In long-term cultures, PRP and FBS did not significantly differ in cell growth rate but differed in morphology and in the expression of vimentin, fibronectin, and &agr;-actin. CONCLUSION: The PRP enhances cell proliferation over the short term and induces cell differentiation of fibroblast-like cells to myofibroblast-like cells over the long term, suggesting that fibroblast differentiation to myofibroblasts may underlie the action mechanism of PRP in soft tissue regeneration.

Bisphosphonate Modulation of the Gene Expression of Different Markers Involved in Osteoblast Physiology: Possible Implications in Bisphosphonate-Related Osteonecrosis of the Jaw

The aim of the present study was to elucidate the role of osteoblasts in bisphosphonates-related osteonecrosis of the jaw (BRONJ). The specific objective was to evaluate the effect on osteoblasts of two nitrogen-containing BPs (zoledronate and alendronate) and one non-nitrogen-containing BP (clodronate) by analyzing modulations in their expression of genes essential for osteoblast physiology. Real-time polymerase chain reaction (RT-PCR) was used to study the effects of zoledronate, alendronate, and clodronate at doses of 10-5, 10-7, or 10-9 M on the expression of Runx-2, OSX, ALP, OSC, OPG, RANKL, Col-I, BMP-2, BMP-7, TGF-β1, VEGF, TGF-βR1, TGF-βR2, and TGF-βR3 by primary human osteoblasts (HOBs) and MG-63 osteosarcoma cells. Expression of these markers was found to be dose-dependent, with no substantive differences between these cell lines. In general, results demonstrated a significant increase in TFG-β1, TGF-βR1, TGF-βR2, TGF-βR3, and VEGF expressions and a significant reduction in RUNX-2, Col-1, OSX, OSC, BMP-2, BMP-7, ALP, and RANKL expressions, while OPG expression varied according to the dose and cell line. The results of this in vitro study of HOBS and MG-63 cell lines indicate that low BP doses can significantly affect the expression of genes essential for osteoblast growth and differentiation and of genes involved in regulating osteoblast-osteoclast interaction, possibly by increasing TGF-β1 production. These findings suggest that osteoblasts may play an important role in BRONJ development, without ruling out other factors.

Cultured Human Fibroblast Biostimulation Using a 940 nm Diode Laser

Background: Fibroblasts are the main cells involved in regeneration during wound healing. The objective was to determine the effect of 940 nm diode laser on cultured human fibroblasts using different irradiation regimens. Methods: The CCD-1064Sk human epithelial fibroblast cell line was treated with a 940 nm diode laser at different energy doses (power: 0.2–1 W and energy density: 1–7 J/cm2) using different transmission modes (continuous or pulsed). The effect on cell growth at 24 and 72 h post-treatment was examined by measuring the proliferative capacity, the impact on the cell cycle, and the effect on cell differentiation. Results: fibroblast proliferative capacity was increased at 24 and 72 h post-treatment as a function of the energy dose. The greatest increase was observed with a power of 0.2 or 0.5 W and energy density between 1 and 4 J/cm2; no difference was observed between continuous and pulsed modes. There were no significant differences in cell cycle between treated groups and controls. α-actin expression was increased by treatment, indicating enhanced cell differentiation. Conclusion: The 940 nm diode laser has biostimulating effects on fibroblasts, stimulating proliferative capacity and cell differentiation without altering the cell cycle. Further researches are necessary to explore its potential clinical usefulness in wound healing.

Effect of phenolic extracts from different extra-virgin olive oil varieties on osteoblast-like cells

The reported incidence of osteoporosis is lower in countries in which the Mediterranean diet predominates, and this apparent relationship may be mediated by the phenolic compounds present in olive oil. The objective of this study was to determine the effect of phenolic extracts from different varieties of extra-virgin olive oil (Picual, Arbequina, Picudo, and Hojiblanca) on the differentiation, antigenic expression, and phagocytic capacity of osteoblast-like MG-63 cells. At 24 h of treatment a significant increase in phosphatase alkaline activity and significant reductions in CD54, CD80, and HLA-DR expression and in phagocytic activity were observed in comparison to untreated controls. The in vitro study performed has demonstrated that phenolic compounds from different extra virgin olive oil varieties can modulate different parameters related to osteoblast differentiation and function.