Hemocompatibility Testing (ISO 10993-4)
Medical devices that come into direct or indirect contact with the blood-circulating system require a specialized set of tests to ensure they do not induce adverse hematological reactions. These interactions can lead to life-threatening complications, such as thrombosis, localized inflammation, or the destruction of red blood cells.
At EBI, we provide comprehensive hemocompatibility evaluations in accordance with ISO 10993-4. Our laboratory utilizes fresh human blood to simulate clinical conditions as closely as possible, ensuring the highest predictive value for patient safety.
Our Hemocompatibility Assays
Depending on your device’s contact duration and category (e.g., blood-path communicating, external communicating, or implants), we offer the following validated assays:
1. Hemolysis (Direct & Indirect)
This test evaluates whether a device or its leachables cause the rupture of red blood cells (erythrocytes), leading to the release of free hemoglobin.
- Methodology: We perform both Direct Contact (blood in contact with the device) and Indirect Contact (blood in contact with the device extract). Diluted human blood with a hemoglobin concentration of 10 ± 1 mg/mL is exposed to the test item. The samples are incubated for 3 hours and subsequently centrifuged.
- Analysis & Evaluation: After exposure, we measure the concentration of free hemoglobin in the supernatant using spectrophotometric methods. Based on free hemoglobin measurements, the blank-corrected hemolytic index (%) is calculated and the hemolytic classification of the material is determined in accordance with ASTM F756.
- Turnaround Time: 6 weeks.
2. Coagulation (PTT test)
This assay evaluates whether the test material activates or interferes with the blood coagulation cascade, which could lead to emboli, thrombus formation or vessel occlusion.
- Methodology: Human plasma is exposed to the test item. The partial thromboplastin time (PTT) is then measured and compared to control samples.
- Analysis & Evaluation: The coagulation response of the test item is evaluated by comparing PTT values to those of a negative control and a blank, in order to determine any shortening or prolongation of clotting time indicative of procoagulant or anticoagulant effects.
- Turnaround Time: 6 weeks.
3. Platelet Activation
Activated platelets are pro-thrombogenic and can lead to the formation of blood clots on the surface of the device.
- Methodology: Human full blood is exposed to the test item. The concentration of CXCL7/PBP (β-thromboglobulin), a marker of platelet activation, is measured in human plasma using ELISA.
- Analysis & Evaluation: Absorbance results are converted into CXCL7/PBP concentrations and compared with the blank and negative control to assess platelet activation.
- Turnaround Time: 6 weeks.
4. Complement Activation
The complement system is a part of the immune system that, if over-activated by a foreign material, can cause a severe inflammatory reaction.
- Methodology: The test item is exposed to human serum. The concentration of the SC5b-9 terminal complement complex (or the marker C3a) is measured using ELISA.
- Analysis & Evaluation: Absorbance results are converted into SC5b-9 concentrations and compared with the blank and negative control to assess complement activation.
- Turnaround Time: 6 weeks.
5. Hematology
A general hematological evaluation is performed to assess the impact of the medical device on blood cell populations, including white blood cells (WBC), red blood cells (RBC), and platelets (PLT), and hematological parameters such as hemoglobin concentration (HGB) and hematocrit (HCT)
- Methodology: Human full blood is exposed to the test item. After completion of the incubation of the test materials with whole human blood, hematological parameters are analyzed using a hematology analyzer.
- Analysis & Evaluation: The results obtained for the hematological parameters WBC, RBC, PLT, HCT, and HGB are compared with the negative control and the blank to assess potential hematological changes induced by contact of blood with the test item.
- Turnaround Time: 6 weeks.
Sample Requirements
Samples must be prepared according to ISO 10993-12 and handled with care to avoid any surface contamination that could influence blood-material interactions.
| Test Type | Material Thickness | Surface Area Required* | Weight / Volume | Minimum Quantity |
|---|---|---|---|---|
| Hemolysis | < 0.5 mm | 30 cm² | - | 6 items |
| ≥ 0.5 mm | 15 cm² | - | 6 items | |
| Liquid / Powder | - | 2 g / 10 ml | - | |
| Coagulation / Platelet activation / Complement activation / Hematology | < 0.5 mm | 30 cm² | - | 3 items |
| ≥ 0.5 mm | 15 cm² | - | 3 items | |
| Liquid / Powder | - | 2 g / 10 ml | - |
*If a volume greater than 30 mL of serum/plasma/blood per test item is required (< 0.5 mm – 180 cm², ≥ 0.5 mm – 90 cm²), representative samples will be taken in accordance with Clause 9.1 of ISO 10993-12:2021. To obtain these representative samples, the test item may need to be cut.
Compliance and Expert Consultation
ISO 10993-4 is one of the most complex parts of the biological evaluation series. Selecting the wrong test category can lead to regulatory rejection. Our team helps you navigate:
- Selection of Blood Donors: Blood is collected from healthy, qualified volunteers in accordance with standardized collection procedures.
- Static vs. Dynamic Testing: Advice on whether your device requires flow-based (dynamic) testing to better simulate vascular conditions.
- Regulatory Submissions: Reports designed for seamless review by Notified Bodies and the FDA.
How We Work
Discovery & Insight
We begin with an in-depth consultation to understand your needs, challenges, and objectives. This helps us define the right strategy and scientific approach for your project.
Design & Development
Our experts translate your vision into actionable plans — combining innovative research, data, and technology to build precise and reliable solutions.
Delivery & Support
We implement the solution with full transparency and ongoing communication. After delivery, we stay involved — supporting you with optimization, updates, and future improvements.
FAQ: Hemocompatibility Best Practices
Why do you use human blood instead of animal blood?
Human blood is the most relevant medium for predicting human clinical outcomes. Significant differences exist in the coagulation and complement systems between species; therefore, ISO 10993-4 recommends using human blood for high-risk devices.
What is the difference between direct and indirect hemolysis?
Direct hemolysis measures the effect of the physical surface of the device on blood cells. Indirect hemolysis measures the effect of chemical substances that might leach out of the device into a solution. Both are usually required for a complete profile.
Which devices require hemocompatibility testing?
Any device that has contact with blood, either directly (heart valves, catheters, stents) or indirectly (infusion sets, blood bags, dialysis equipment), must undergo hemocompatibility assessment.
Why do you use human blood instead of animal blood?
Human blood is the most relevant medium for predicting human clinical outcomes. Significant differences exist in the coagulation and complement systems between species; therefore, ISO 10993-4 recommends using human blood for high-risk devices.
What is the difference between direct and indirect hemolysis?
Direct hemolysis measures the effect of the physical surface of the device on blood cells. Indirect hemolysis measures the effect of chemical substances that might leach out of the device into a solution. Both are usually required for a complete profile.
Which devices require hemocompatibility testing?
Any device that has contact with blood, either directly (heart valves, catheters, stents) or indirectly (infusion sets, blood bags, dialysis equipment), must undergo hemocompatibility assessment.
Contact with us
We’re open to collaboration and research partnerships. Fill out the form — our team will respond as soon as possible.
Location:
Europejski Instytut Biomedyczny sp. z o.o.
ul. Bartycka 63A lok. 6, 00-716 Warszawa
Mail:
contact@ebi.bio
Phone:
+48 22 780 06 32
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