ArticleMethods and protocols2020
Combined Fluorescence-Based in Vitro Assay for the Simultaneous Detection of Cell Viability and Alkaline Phosphatase Activity during Osteogenic Differentiation of Osteoblast Precursor Cells.
Article in Methods and protocols, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Hydrogel and Injectable Platelet-Rich Fibrin: A Synergistic Approach to Osteogenesis.European journal of dentistry · 2026Article
- Impact of 45S5-Bioactive Glass on Chondrocytes in Knee Osteoarthritis-In Vitro Study Exploring Cellular Responses.Journal of functional biomaterials · 2025Article
- Exploring the mechanical strength, antimicrobial performance, and bioactivity of 3D-printed silicon nitride-PEEK composites in cervical spinal cages.International journal of bioprinting · 2024Article
- The Impact of 45S5-Bioactive Glass on Synovial Cells in Knee Osteoarthritis-An In Vitro Study.Materials (Basel, Switzerland) · 2023Article
- In Vitro Evaluation of the Effects of Hyaluronic Acid and an Aminoacidic Pool on Human Osteoblasts.Biomedicines · 2023Article
- Strontium doped bioglass incorporated hydrogel-based scaffold for amplified bone tissue regeneration.Scientific reports · 2022Article
- Synthesis, Structure-Property Evaluation and Biological Assessment of Supramolecular Assemblies of Bioactive Glass with Glycyrrhizic Acid and Its Monoammonium Salt.Materials (Basel, Switzerland) · 2022Article
- In vitro and in ovo impact of the ionic dissolution products of boron-doped bioactive silicate glasses on cell viability, osteogenesis and angiogenesis.Scientific reports · 2022Article
- Inhibitory Effect ofMedicina (Kaunas, Lithuania) · 2022Article
- An In Vitro Evaluation of the Biological and Osteogenic Properties of Magnesium-Doped Bioactive Glasses for Application in Bone Tissue Engineering.International journal of molecular sciences · 2021Article
- Staphylococcus aureus internalization impairs osteoblastic activity and early differentiation process.Scientific reports · 2021Article
- Impact of Zinc- or Copper-Doped Mesoporous Bioactive Glass Nanoparticles on the Osteogenic Differentiation and Matrix Formation of Mesenchymal Stromal Cells.Materials (Basel, Switzerland) · 2021Article
- Preconditioning of Bioactive Glasses before Introduction to Static Cell Culture: What Is Really Necessary?Methods and protocols · 2020Article
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3 authors.
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Abstract
Novel bone substitute materials need to be evaluated in terms of their osteogenic differentiation capacity and possible unwanted cytotoxic effects in order to identify promising candidates for the therapy of bone defects. The activity of alkaline phosphatase (ALP) is frequently quantified as an osteogenic marker, while various colorimetric assays, like MTT assay, are used to monitor cell viability. In addition, the DNA or protein content of the samples needs to be quantified for normalization purposes. As this approach is time consuming and often requires the analysis of multiple samples, we aimed to simplify this process and established a protocol for the combined fluorescence-based quantification of ALP activity and cell viability within one single measurement. We demonstrate that the fluorogenic substrate 4-methylumbelliferone-phosphate (4-MUP) and the commonly used para-nitrophenylphosphate (p-NPP) produce comparable and highly correlating results. We further show that fluorescein-diacetate (FDA) can be used to quantify both cell viability and cell number without interfering with the quantification of ALP activity. The measurement of additional normalization parameters is, therefore, unnecessary. Therefore, the presented assay allows for a time-efficient, simple and reliable analysis of both ALP activity and cell viability from one sample and might facilitate experiments evaluating the osteogenic differentiation of osteoblast precursor cells.
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