In search of an osteoblast cell model for in vitro research

Czekanska, Ewa M.; Stoddart, M J; Richards, Rebecca; Hayes, J. S.

doi:10.22203/ecm.v024a01

Cited by 478 publications

(449 citation statements)

References 144 publications

(148 reference statements)

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“…However, comparability can only be achieved to a limited extent. Isolated human tissue is always subject to donor-related individual variations regarding age, gender and hormone levels [37]. Additionally, the in vitro osteoblast cell culture does not offer all functions of a human bone tissue.…”

Section: Discussionmentioning

confidence: 99%

Human osteoblast damage after antiseptic treatment

Vörös

Dobrindt

Perka

et al. 2013

International Orthopaedics (SICOT)

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Purpose Antiseptics are powerful medical agents used for wound treatment and decontamination and have a high potential for defeating joint infections in septic surgery. Both chlorhexidine and polyhexanide are frequently used in clinical practice and have a broad antimicrobial range, but their effect on human osteoblasts has not been sufficiently studied. Our objective was to investigate the toxic effects of polyhexanide and chlorhexidine on human osteoblasts in vitro to evaluate their clinical applicability in septic surgery. Methods We isolated and cultivated human osteoblasts in vitro and assayed the toxic effects of chlorhexidine 0.1 % and polyhexanide 0.04 %, concentrations commonly applied in clinical practice. Toxicity analysis was performed by visualisation of cell structure, lactate dehydrogenase (LDH) activity and evaluation of vital cells. Toxicity was evaluated by microscopic inspection of cell morphology, trypan blue staining and determination of LDH release. Results Damaged cell structure could be shown by microscopy. Both antiseptics promoted LDH activity after incubation with osteoblasts. The evaluation of vital osteoblasts showed a significant decrease of vital cells. Conclusions Both antiseptics induced significant cell death of osteoblasts at optimum exposure. We therefore recommend cautious use of polyhexanide and chlorhexidine in septic surgery to avoid severe osteoblast toxicity.

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Section: Discussionmentioning

confidence: 99%

Human osteoblast damage after antiseptic treatment

Vörös

Dobrindt

Perka

et al. 2013

International Orthopaedics (SICOT)

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“…hOB and hMSC were seeded at 10,000 cells/ cm 2 in growth media (ScienCell) and proliferation was measured at 48 hours per the manufacturer's instructions. Osteogenic differentiation was evaluated on each surface using clonal mouse preosteoblast cells (MC3T3-E1; ATCC, Manassas, VA, USA) as a result of their homogeneity, availability, and differentiation profile that is more similar to human osteoblasts than other in vitro models [7]. MC3T3 cells were seeded at 20,000 cells/cm 2 in growth media composed of a-MEM (Life Technologies, Carlsbad, CA, USA) supplemented with 16.7% fetal bovine serum (Atlanta Biologicals, Lawrenceville, GA, USA) and 1% penicillin-streptomycin-L-glutamine (Life Technologies).…”

Section: Methodsmentioning

confidence: 99%

Do Surface Porosity and Pore Size Influence Mechanical Properties and Cellular Response to PEEK?

Torstrick

Evans

Stevens

et al. 2016

Clinical Orthopaedics &Amp; Related Research

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Background Despite its widespread use in orthopaedic implants such as soft tissue fasteners and spinal intervertebral implants, polyetheretherketone (PEEK) often suffers from poor osseointegration. Introducing porosity can overcome this limitation by encouraging bone ingrowth; however, the corresponding decrease in implant strength can potentially reduce the implant's ability to bear physiologic loads. We have previously shown, using a single pore size, that limiting porosity to the surface of PEEK implants preserves strength while supporting in vivo osseointegration. However, additional work is needed to investigate the effect of pore size on both the mechanical properties and cellular response to PEEK. Questions/purposes (1) Can surface porous PEEK (PEEK-SP) microstructure be reliably controlled? (2) What is the effect of pore size on the mechanical properties of PEEK-SP? (3) Do surface porosity and pore size influence the cellular response to PEEK? Methods PEEK-SP was created by extruding PEEK through NaCl crystals of three controlled ranges: 200 to 312, 312 to 425, and 425 to 508 lm. Micro-CT was used to characterize the microstructure of PEEK-SP. Tensile, fatigue, and interfacial shear tests were performed to compare the mechanical properties of PEEK-SP with injectionmolded PEEK (PEEK-IM). The cellular response to PEEK-SP, assessed by proliferation, alkaline phosphatase activity, vascular endothelial growth factor production, and calcium content of osteoblast, mesenchymal stem cell, and preosteoblast (MC3T3-E1) cultures, was compared with that of machined smooth PEEK and Ti6Al4V. Results Micro-CT analysis showed that PEEK-SP layers possessed pores that were 284 ± 35 lm, 341 ± 49 lm, and 416 ± 54 lm for each pore size group. Porosity and

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“…Specifically, these cells produce type I collagen with no or low basal osteocalcin (OC) and ALP. However, when MG63's are treated with 1,25D, OC expression increases [4][5] and, when the same cells are co-treated with 1,25D and selected growth factors, e.g. lysophosphatidic acid, the levels of ALP markedly increase [6], a feature of the mature osteoblast phenotype.…”

Section: Human Osteoblastsmentioning

confidence: 99%

Alkaline phosphatase binds tenaciously to titanium; implications for biological surface evaluation following bone implant retrieval

Mansell

Shiel

Harwood³

et al. 2017

Materials Science and Engineering: C

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Enhancing the performance and longevity of titanium (Ti) implants continues to be a significant developmental theme in contemporary biomaterials design. Our specific focus pertains to the surface functionalisation of Ti using the bioactive lipid, lysophosphatidic acid (LPA) and certain phosphatase-resistant analogues of LPA. Coating survivorship to a plethora of testing regimens is required to align with due regulatory process before novel biomaterials can enter clinical trials. One of the key acceptance criteria is coating retention to the physical stresses experienced during implantation. In assessing coating stability to insertion into porcine bone we found that a subsequent in vitro assessment to confirm coating persistence was masked by abundant alkaline phosphatase (ALP) contamination adsorbed to the metal surface. Herein we report that ALP can bind to Ti in a matter of minutes by simply immersing Ti samples in aqueous solutions of the enzyme. We strongly discourage the in vitro monitoring of osteoblast and stromal cell ALP expression when assessing bioactive coating survivorship following Ti implant retrieval form native bone tissue.

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In search of an osteoblast cell model for in vitro research

Cited by 478 publications

References 144 publications

Human osteoblast damage after antiseptic treatment

Human osteoblast damage after antiseptic treatment

Do Surface Porosity and Pore Size Influence Mechanical Properties and Cellular Response to PEEK?

Alkaline phosphatase binds tenaciously to titanium; implications for biological surface evaluation following bone implant retrieval

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