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Fulltext Alteration of cell cytoskeleton and functions of cell recovery of normal human osteoblast cells caused by factors associated with real space flight

Kapitonova MY, Salim N, Othman S, Muhd Kamauzaman TM, Ali AM, Nawawi HM, et al.

Malays J Pathol, 2013 Dec;35(2):153-63.
PMID: 24362479 MyJurnal

Experiments involving short-term space flight have shown an adverse effect on the physiology, morphology and functions of cells investigated. The causes for this effect on cells are: microgravity, temperature fluctuations, mechanical stress, hypergravity, nutrient restriction and others. However, the extent to which these adverse effects can be repaired by short-term space flown cells when recultured in conditions of normal gravity remains unclear. Therefore this study aimed to investigate the effect of short-term spaceflight on cytoskeleton distribution and recovery of cell functions of normal human osteoblast cells. The ultrastructure was evaluated using ESEM. Fluorescent staining was done using Hoechst, Mito Tracker CMXRos and Tubulin Tracker Green for cytoskeleton. Gene expression of cell functions was quantified using qPCR. As a result, recovered cells did not show any apoptotic markers when compared with control. Tubulin volume density (p<0.001) was decreased significantly when compared to control, while mitochondria volume density was insignificantly elevated. Gene expression for IL-6 (p<0.05) and sVCAM-1 (p<0.001) was significantly decreased while alkaline phosphatase (p<0.001), osteocalcin and sICAM (p<0.05) were significantly increased in the recovered cells compared to the control ones. The changes in gene and protein expression of collagen 1A, osteonectin, osteoprotegerin and beta-actin, caused by short-term spaceflight, were statistically not significant. These data indicate that short term space flight causes morphological changes in osteoblast cells which are consistent with hypertrophy, reduced cell differentiation and increased release of monocyte attracting proteins. The long-term effect of these changes on bone density and remodeling requires more detailed studies.

Matched MeSH terms: Osteoblasts/ultrastructure*
Polydopamine-assisted chlorhexidine immobilization on medical grade stainless steel 316L: Apatite formation and in vitro osteoblastic evaluation

Mohd Daud N, Hussein Al-Ashwal R, Abdul Kadir MR, Saidin S

Ann. Anat., 2018 Nov;220:29-37.
PMID: 30048761 DOI: 10.1016/j.aanat.2018.06.009

Immobilization of chlorhexidine (CHX) on stainless steel 316L (SS316L), assisted by a polydopamine film as an intermediate layer is projected as an approach in combating infection while aiding bone regeneration for coating development on orthopedic and dental implants. This study aimed to investigate the ability of CHX coating to promote apatite layer, osteoblast cells viability, adhesion, osteogenic differentiation and mineralization. Stainless steel 316L disks were pre-treated, grafted with a polydopamine film and immobilized with different concentrations of CHX (10-30mM). The apatite layer formation was determined through an in vitro simulated body fluid (SBF) test by ATR-FTIR and SEM-EDX analyses. The osteoblastic evaluations including cells viability, cells adhesion, osteogenic differentiation and mineralization were assessed with human fetal osteoblast cells through MTT assay, morphology evaluation under FESEM, ALP enzyme activity and Alizarin Red S assay. The apatite layer was successfully formed on the CHX coated disks, demonstrating potential excellent bioactivity property. The CHX coatings were biocompatible with the osteoblast cells at low CHX concentration (<20mM) with good adhesion on the metal surfaces. The increment of ALP activity and calcium deposition testified that the CHX coated disks able to support osteoblastic maturation and mineralization. These capabilities give a promising value to the CHX coating to be implied in bone regeneration area.

Matched MeSH terms: Osteoblasts/ultrastructure*
Microwave-assisted fabrication of titanium implants with controlled surface topography for rapid bone healing

Kutty MG, De A, Bhaduri SB, Yaghoubi A

ACS Appl Mater Interfaces, 2014 Aug 27;6(16):13587-93.
PMID: 25095907 DOI: 10.1021/am502967n

Morphological surface modifications have been reported to enhance the performance of biomedical implants. However, current methods of introducing graded porosity involves postprocessing techniques that lead to formation of microcracks, delamination, loss of fatigue strength, and, overall, poor mechanical properties. To address these issues, we developed a microwave sintering procedure whereby pure titanium powder can be readily densified into implants with graded porosity in a single step. Using this approach, surface topography of implants can be closely controlled to have a distinctive combination of surface area, pore size, and surface roughness. In this study, the effect of various surface topographies on in vitro response of neonatal rat calvarial osteoblast in terms of attachment and proliferation is studied. Certain graded surfaces nearly double the chance of cell viability in early stages (∼one month) and are therefore expected to improve the rate of healing. On the other hand, while the osteoblast morphology significantly differs in each sample at different periods, there is no straightforward correlation between early proliferation and quantitative surface parameters such as average roughness or surface area. This indicates that the nature of cell-surface interactions likely depends on other factors, including spatial parameters.

Matched MeSH terms: Osteoblasts/ultrastructure
Eurycoma longifolia as a potential alternative to testosterone for the treatment of osteoporosis: Exploring time-mannered proliferative, differentiative and morphogenic modulation in osteoblasts

Thu HE, Mohamed IN, Hussain Z, Shuid AN

J Ethnopharmacol, 2017 Jan 04;195:143-158.
PMID: 27818256 DOI: 10.1016/j.jep.2016.10.085

ETHNOPHARMACOLOGICAL RELEVANCE: Eurycoma longifolia (EL) has been well-studied traditionally as a chief ingredient of many polyherbal formulations for the management of male osteoporosis. It has also been well-recognised to protect against bone calcium loss in orchidectomised rats.
AIM OF THE STUDY: To evaluate the effects of EL on the time-mannered sequential proliferative, differentiative, and morphogenic modulation in osteoblasts compared with testosterone.
MATERIALS AND METHODS: Cell proliferation was analysed using MTS assay and phase contrast microscopy. Osteogenic differentiation of MC3T3-E1 cells was assessed through a series of characteristic assays which include crystal violet staining, alkaline phosphatase (ALP) activity and Van Gieson staining. Taken together, the bone mineralization of extra cellular matrix (ECM) was estimated using alizarin red s (ARS) staining, von kossa staining, scanning electron microscopic (SEM) and energy dispersive x-ray (EDX) analysis.
RESULTS: The cell proliferation data clearly revealed the efficiency of EL particularly at a dose of 25µg/mL, in improving the growth of MC3T3-E1 cells compared with the untreated cells. Data also showed the prominence of EL in significantly promoting ALP activity throughout the entire duration of treatment compared with the testosterone-treated cells. The osteogenic differentiation potential of EL was further explored by analysing mineralization data which revealed that the calcified nodule formation (calcium deposition) and phosphate deposition was more pronounced in cells treated with 25µg/mL concentration of EL at various time points compared with the untreated and testosterone treated cells. The scanning electron microscopic (SEM) analysis also revealed highest globular masses of mineral deposits (identified as white colour crystals) in the ECM of cultured cells treated with 25µg/mL concentration of EL.
CONCLUSION: Compared to testosterone, greater potential of EL in promoting the proliferation and osteogenic differentiation of MC3T3-E1 cells provides an in vitro basis for the prevention of male osteoporosis. Thus, we anticipate that EL can be considered as an alternative approach to testosterone replacement therapy (TRT) for the treatment of male osteoporosis.

Matched MeSH terms: Osteoblasts/ultrastructure