Physiological dynamic strain amplitudes increase human osteoblast proliferation but decrease osteocalcin synthesis in vitro

AbstractExperimental methods for studying an enhancement of osteoblast proliferation in vitro provide tools for the research of biochemical processes involved in bone turnover in vivo. Some of the current methods used for this purpose are based on the ability of the osteoblasts to enhance proliferation by mechanical stimulation. We describe an experimental approach of biomechanical stimulation of cultured human osteoblast-like cells by vibration. This method is based on the specially designed controlled vibration setup that consists of an electric actuator, with horizontally mounted well plate containing cell cultures. Previously this method found to be effective to enhance cell proliferation, but the exact mechanical parameters of effective vibration were elusive. The current low friction system for mechanical stimulation of osteoblast-like cells in vitro provides recording of narrow range mechanical parameters in the infrasonic spectrum.We exposed human osteoblast-like cells in explant monolayer culture to mechanical vibration in the 10-70Hz range of frequencies and found that 50-70 Hz of vibration frequency is optimal for inducing osteoblast proliferation that was deduced from interrelation between unchanged cell number in culture samples with significant decrease in cell death rate (decreased LDH activity in culture media, p<.05) and with parallel decrease of their maturation level (p<.01).In this report we determined the optimal mechanical parameters and excitation protocol for induction of osteoblast proliferation in vitro by using a tunable and versatile mechanical platform, which can be used in the research of cell mechanotransduction.

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Optimal parameters for the enhancement of human osteoblast-like cell proliferation in vitro via shear stress induced by high-frequency mechanical vibration

Iberoamerican Journal of Medicine ◽

10.53986/ibjm.2021.0033 ◽

2021 ◽

Vol 3 (3) ◽

pp. 204-211

Author(s):

Nahum Rosenberg ◽

◽

Orit Rosenberg ◽

Jacob Halevi Politch ◽

Haim Abramovich ◽

...

Keyword(s):

Cell Proliferation ◽

Cell Death ◽

Monolayer Culture ◽

Mechanical Vibration ◽

Mechanical Parameters ◽

Human Osteoblast ◽

Osteoblast Proliferation ◽

Static Conditions ◽

Proliferation In Vitro

Introduction: Biomechanical stimulation of cultured human osteoblast-like cells, which is based on controlled mechanical vibration, has been previously indicated, but the exact mechanical parameters that are effective for cells' proliferation enhancement are still elusive due to the lack of direct data recordings from the stimulated cells in culture. Therefore, we developed a low friction tunable system that enables recording of a narrow range of mechanical parameters, above the infrasonic spectrum, that applied uniformly to human osteoblast-like cells in monolayer culture, aiming to identify a range of mechanical parameters that are effective to enhance osteoblast proliferation in vitro. Methods: Human osteoblast-like cells in explant monolayer culture samples were exposed to mechanical vibration in the 10-70Hz range of frequencies for two minutes, in four 24 hours intervals. Cell numbers in culture, cellular alkaline phosphatase activity (a marker of cell maturation), and lactate dehydrogenase activity in culture media (representing cell death) were measured after the mechanical stimulation protocol application and compared statistically to the control cell cultures kept in static conditions. The cell proliferation was deduced from cell number in culture and cell death measurements. Results: We found that 50-70 Hz of vibration frequency protocol (10-30 μm of maximal displacement amplitude, 0.03g of peak-to-peak acceleration) is optimal for enhancing cells' proliferation(p<0.05), with a parallel decrease of their maturation (p<0.01). Discussion: We detected the optimal mechanical parameters of excitation protocol for induction of osteoblast proliferation in vitro by a mechanical platform, which can be used as a standardized method in the research of mechanotransduction in human osteoblast.

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L-Carnitine and Isovaleryl L-Carnitine Fumarate Positively Affect Human Osteoblast Proliferation and Differentiation In Vitro

Calcified Tissue International ◽

10.1007/s00223-004-0147-4 ◽

2005 ◽

Vol 76 (6) ◽

pp. 458-465 ◽

Cited By ~ 31

Author(s):

S. Colucci ◽

G. Mori ◽

S. Vaira ◽

G. Brunetti ◽

G. Greco ◽

...

Keyword(s):

Human Osteoblast ◽

Osteoblast Proliferation ◽

Proliferation And Differentiation

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In vitro study of human osteoblast proliferation and morphology on orthodontic mini-implants

The Angle Orthodontist ◽

10.2319/100714-717.1 ◽

2015 ◽

Vol 85 (6) ◽

pp. 920-926 ◽

Cited By ~ 2

Author(s):

Ricardo Carvalho Bueno ◽

Roberta Tarkany Basting

Keyword(s):

Chemical Composition ◽

In Vitro Study ◽

Chemical Components ◽

Control Group ◽

Osteoblast Proliferation ◽

Mini Implants ◽

Significant Difference ◽

Implant Anchorage ◽

Positive Effect

ABSTRACT Objective: To evaluate the proliferation and morphology of human osteoblasts cultured on two brands of mini-implants after 24, 48, and 72 hours, in addition to the chemical composition found on their surface. Materials and Methods: Two brands of mini-implant (Morelli and Neodent) were evaluated; polystyrene was used as a control group (n = 3). Osteoblasts were cultured on the surface of sterilized mini-implants in a CO2 incubator at different time periods (24, 48, and 72 hours). Osteoblast proliferation was quantified by scanning electron microscopy using up to 5000× magnification, and cell morphology was analyzed by a single observer. For the chemical analysis, spectroscopy X-ray fluorescence was used to identify and quantify chemical components on the surface of the mini-implants. Results: Two-way ANOVA showed no significant interaction between the factors studied (P = 0.686). A Tukey test revealed no significant difference in osteoblast proliferation between the mini-implants at all studied periods; however, a difference in cell proliferation was detected between the Neodent and the control group (P = .025). For all groups, time had a direct and positive effect on osteoblast proliferation (P < .001). The significant elements present in both brands of mini-implants were titanium, aluminum, vanadium, and iron. Conclusions: Osteoblast proliferation was present on the mini-implants studied, which increased over time; however, no significant difference between brands was observed. No difference was seen between the mini-implants evaluated in terms of chemical composition. Cell adhesion after 72 hours suggests that areas of bone remodeling can be achieved, thus initiating the process of mini-implant anchorage.

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In vitro catabolic effect of protoporphyrin IX in human osteoblast-like cells: possible role of the 18 kDa mitochondrial translocator protein

Journal of Bioenergetics and Biomembranes ◽

10.1007/s10863-013-9501-4 ◽

2013 ◽

Vol 45 (4) ◽

pp. 333-341 ◽

Cited By ~ 10

Author(s):

Nahum Rosenberg ◽

Orit Rosenberg ◽

Abraham Weizman ◽

Leo Veenman ◽

Moshe Gavish

Keyword(s):

Protoporphyrin Ix ◽

Translocator Protein ◽

Human Osteoblast

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THE EFFECTS OF PARTICULATE COBALT, CHROMIUM AND COBALT-CHROMIUM ALLOY ON HUMAN OSTEOBLAST-LIKE CELLS IN VITRO

Journal of Bone and Joint Surgery - British Volume ◽

10.1302/0301-620x.79b3.0790475 ◽

1997 ◽

Vol 79-B (3) ◽

pp. 475-482 ◽

Cited By ~ 35

Author(s):

M. J. Allen ◽

B. J. Myer ◽

P. J. Millett ◽

N. Rushton

Keyword(s):

Chromium Alloy ◽

Human Osteoblast ◽

Cobalt Chromium ◽

Cobalt Chromium Alloy

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Antioxidative 1,4-Dihydropyridine Derivatives Modulate Oxidative Stress and Growth of Human Osteoblast-Like Cells In Vitro

Antioxidants ◽

10.3390/antiox7090123 ◽

2018 ◽

Vol 7 (9) ◽

pp. 123 ◽

Cited By ~ 8

Author(s):

Lidija Milkovic ◽

Tea Vukovic ◽

Neven Zarkovic ◽

Franz Tatzber ◽

Egils Bisenieks ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Growth ◽

Cell Model ◽

Second Messengers ◽

Human Osteoblast ◽

Tert Butyl Hydroperoxide ◽

Human Stress ◽

Set Up ◽

Dihydropyridine Derivatives

Oxidative stress has been implicated in pathophysiology of different human stress- and age-associated disorders, including osteoporosis for which antioxidants could be considered as therapeutic remedies as was suggested recently. The 1,4-dihydropyridine (DHP) derivatives are known for their pleiotropic activity, with some also acting as antioxidants. To find compounds with potential antioxidative activity, a group of 27 structurally diverse DHPs, as well as one pyridine compound, were studied. A group of 11 DHPs with 10-fold higher antioxidative potential than of uric acid, were further tested in cell model of human osteoblast-like cells. Short-term combined effects of DHPs and 50 µM H2O2 (1-h each), revealed better antioxidative potential of DHPs if administered before a stressor. Indirect 24-h effect of DHPs was evaluated in cells further exposed to mild oxidative stress conditions induced either by H2O2 or tert-butyl hydroperoxide (both 50 µM). Cell growth (viability and proliferation), generation of ROS and intracellular glutathione concentration were evaluated. The promotion of cell growth was highly dependent on the concentrations of DHPs used, type of stressor applied and treatment set-up. Thiocarbatone III-1, E2-134-1 III-4, Carbatone II-1, AV-153 IV-1, and Diethone I could be considered as therapeutic agents for osteoporosis although further research is needed to elucidate their bioactivity mechanisms, in particular in respect to signaling pathways involving 4-hydroxynoneal and related second messengers of free radicals.

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