Comprehensive in vitro services to characterize nanoparticle–biological interactions and accelerate selection.
Bridging the gap between nanoparticle synthesis and in vivo studies requires rigorous in vitro assessment. Understanding how nanomaterials interact with cellular environments, blood components, and immune systems is crucial for optimizing formulation and reducing attrition rates. BOC Sciences offers a suite of specialized cell-based and cell-free assays to evaluate the biocompatibility, efficacy, and stability of your nanocarriers. From standard cytotoxicity screening to complex mechanistic studies involving intracellular trafficking and protein corona analysis, we provide high-quality data to support your research and development decisions.
In vitro safety profiling of nanoparticle formulationsWe provide a versatile platform for evaluating lipid nanoparticles, polymeric nanoparticles, inorganic particles, and exosomes. Our assays are optimized to minimize interference often caused by nanomaterial optical properties.
Assessment of acute and metabolic toxicity across a wide library of cancer and normal cell lines. We employ multiple assay formats to avoid false positives common with nanomaterials.
Quantitative and qualitative analysis of how nanoparticles enter cells and where they localize. Crucial for confirming payload delivery to the cytoplasm or nucleus.
Critical evaluation for nanoparticles intended for intravenous administration. We test interactions with blood components to predict circulation stability and safety.
Investigation of inflammatory responses and cellular stress mechanisms induced by nanomaterials, ensuring the material does not trigger unwanted immune reactions.
Characterization of payload retention and release kinetics under simulated physiological conditions, including serum-containing environments.
Determining the mechanism of cell death and the impact of treatment on cell division, essential for validating anti-cancer nanotherapeutics.
Secure the biological proof-of-concept for your nanomaterials. Our comprehensive evaluation reports provide the insight needed to optimize formulations before moving to animal studies.
Our facility is equipped with industry-standard biological instrumentation tailored for high-sensitivity detection of nanoparticle effects in complex biological matrices.
| Instrument / Platform | Key Capabilities & Applications |
|---|---|
| I. Cellular Analysis & Imaging | |
| Confocal Laser Scanning Microscope (CLSM) | Z-stack imaging for 3D localization of nanoparticles; Co-localization analysis with organelle markers; Live-cell imaging. |
| Flow Cytometer | Quantitative cellular uptake analysis; Cell cycle distribution; Apoptosis detection (Annexin V); Surface marker expression. |
| High-Content Screening System | Automated imaging and analysis for multi-parametric cytotoxicity and morphological changes in high-throughput formats. |
| II. Spectroscopic & Plate-Based Assays | |
| Multimode Microplate Reader | Fluorescence, Luminescence, and Absorbance modes for ELISA, cytotoxicity (CCK-8/MTT), and ROS assays. |
| UV-Vis Spectrophotometer | Hemolysis quantification; Drug release monitoring; Turbidity measurements for aggregation studies. |
| III. Molecular Biology & Biochemistry | |
| Real-Time PCR (qPCR) | Gene expression analysis to evaluate inflammatory responses or gene silencing efficiency (for siRNA/mRNA NPs). |
| Gel Electrophoresis & Western Blot | Protein analysis for apoptosis markers (Caspases, PARP) and signaling pathway activation; siRNA integrity analysis. |
Standard biological assays often fail when applied to nanomaterials due to unique physical properties. We employ specialized protocols to ensure data accuracy.
✔ Mitigating Optical Interference
Many nanoparticles absorb at detection wavelengths (e.g., Gold NPs, Carbon nanotubes). We use appropriate background controls and alternative non-optical assays (e.g., LDH vs MTT) to prevent false toxicity readings.
✔ Analyzing Hydrophobic Drugs
Evaluating the release of poorly soluble drugs is challenging. We utilize "sink conditions" with surfactants or serum proteins to realistically simulate in vivo release profiles without saturation artifacts.
✔ Serum Stability & Aggregation
Nanoparticles stable in water may aggregate in culture media. We characterize size distribution (DLS) in relevant biological media before dosing to ensure cells are exposed to dispersed particles, not aggregates.
✔ Distinguishing Uptake vs. Adsorption
Particles stuck to the cell surface can mimic uptake. We use Trypan Blue quenching or acid washing techniques during FACS analysis to differentiate internalized particles from surface-bound ones.
✔ Endotoxin Control
Contamination with LPS (endotoxin) can trigger immune responses falsely attributed to the nanoparticle. We offer endotoxin screening (LAL assay) to ensure observed immunotoxicity is material-intrinsic.
✔ Relevant Control Selection
We include appropriate positive (e.g., known toxic compounds) and negative controls, as well as vehicle controls, to statistically validate the biological effects observed in your study.
We define the cell lines, endpoints (toxicity, uptake, release), and controls based on your nanoparticle type and therapeutic indication.
Nanoparticles are sterilized (if possible) and dispersed in culture media. Cells are treated according to dose-response and time-course protocols.
Use FACS, microscopy, and plate readers to measure cytotoxicity, cellular uptake, intracellular localization, and release kinetics; statistically analyze IC50, efficiency, and functional effects.
Delivery of a comprehensive research report including raw data, images, graphs, and a summary of biological findings suitable for publication or internal review.
Client: A biotech startup developing mRNA vaccines.
Challenge: High cellular uptake was observed, yet protein expression remained unexpectedly low, suggesting that the mRNA cargo was being trapped and degraded within the endo-lysosomal pathway.
Solution: A rigorous co-localization study was implemented using high-resolution CLSM to track the intracellular fate of the particles. Cells were treated with fluorescently labeled mRNA-LNPs, while lysosomes were precisely demarcated with LysoTracker dyes. The evaluation involved a side-by-side comparison between the client's standard formulation and a newly engineered pH-sensitive lipid variant, utilizing advanced image analysis software to quantify the degree of overlap between the carrier and the degradative compartments over a 24-hour time course.
Outcome: Quantitative image analysis revealed a significant reduction in the Pearson correlation coefficient for the new formulation, confirming successful endosomal escape. This breakthrough directly correlated with a 5-fold increase in reporter protein expression.
Client: Academic group researching silver nanoparticles for antimicrobial blood therapies.
Challenge: Detailed safety data regarding potential red blood cell (RBC) lysis was required to mitigate risks before proceeding to systemic administration in rodent models.
Solution: A comprehensive hemocompatibility assessment was executed, featuring a full panel of blood-interaction assays. This involved testing hemolysis rates across an extensive concentration gradient (1 to 100 μg/mL) using fresh human blood components to ensure physiological relevance. Furthermore, the study integrated coagulation cascade analysis, measuring Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT), alongside platelet aggregation studies to identify any subtle thrombogenic potential or interference with hematological homeostasis.
Outcome: The evaluation successfully defined a safe therapeutic window. While high concentrations showed 15% hemolysis, levels below 10 μg/mL maintained hemolysis at <2%, ensuring no adverse impact on coagulation during subsequent in vivo trials.
Customized Assay DesignWe do not rely on "one-size-fits-all" protocols. We tailor cell models (2D vs 3D, specific cell lines) and assay conditions to mimic the specific biological barriers your nanoparticle must overcome.
Nanomaterial ExpertiseUnlike general CROs, we specialize in nanomaterials. We understand colloidal stability, protein corona effects, and optical interference, ensuring our data reflects true biological interactions.
Comprehensive Data ReportingOur reports serve as standalone research documents, providing detailed methodology, raw data, statistical analysis, and high-resolution microscopy images suitable for grant applications and publications.
Extensive Cell BankAccess to a wide variety of human and animal cell lines, including cancer lines (HeLa, MCF-7, A549), immune cells (RAW264.7, THP-1), and normal fibroblasts/epithelial cells for toxicity comparisons.
Integrated Service PipelineSeamlessly transition from our nanoparticle synthesis and conjugation services directly to in vitro evaluation, minimizing logistics and sample degradation risks.
Oxidative stress is a common response to nanoparticle exposure. We implement ROS detection assays, glutathione depletion measurements, and antioxidant enzyme activity tests to evaluate cellular oxidative responses. By integrating these assays, BOC Sciences provides detailed mechanistic insights, enabling clients to identify potentially reactive nanoparticle formulations and optimize safety profiles at the in vitro stage.
Cellular internalization is a key determinant of nanoparticle performance. We utilize fluorescence microscopy, flow cytometry, and confocal imaging to track nanoparticle entry, localization, and accumulation within cells. Our team can customize labeling strategies and time-course studies to deliver quantitative and qualitative insights, helping clients understand mechanisms and improve nanoparticle design.
Cytotoxicity evaluation is critical for understanding nanoparticle-cell interactions. We employ a range of assays, including MTT, LDH, and live/dead staining, to quantify cell viability and membrane integrity. Our platform allows flexible selection of cell lines and exposure conditions, enabling precise assessment of dose-dependent effects and biocompatibility, providing clients with robust data to guide nanoparticle optimization.
Nanoparticle-membrane interactions influence uptake and toxicity. Techniques such as hemolysis assays, membrane permeability studies, and lipid bilayer modeling are applied to quantify these effects. Our team can simulate various cellular environments and nanoparticle concentrations, delivering actionable data that guide structural modifications to enhance compatibility and functional performance.
Inflammatory responses can impact nanoparticle application. We assess cytokine secretion, NF-κB activation, and immune cell signaling in relevant in vitro models to detect pro-inflammatory effects. BOC Sciences’ integrated evaluation provides a comprehensive profile of immune interactions, supporting clients in selecting formulations that minimize adverse cellular responses while maintaining desired functionality.
