Biological Materials Analysis

In-vitro inflammation models for drug development

The group is developing macrophage-based, standardized inflammation models for drug development, which enable the investigation of various inflammatory and immunomodulatory mechanisms. The models are adapted according to the intended use of the drugs.

The in-vitro test scenarios to be developed for the preclinical testing of active pharmaceutical ingredients are 1) drug screening assays, 2) mode-of-action assays, 3) bioassays (potency assays) and 4) safety analyses.

Assay development for medical products for service and transfer

• Test development for proof of the efficacy, the immunomodulatory potential, and off-target effects of APIs or substances
• Test development for proof of the immunomodulatory potential of material extracts or in direct cell-material contact
• Test development for an immunotoxicological investigation (analogous to ISO 10993-10, -20 and -22), i.a. acute inflammation assay as part of the immunotoxicity
• Testing of the cytotoxicity/cell viability of substances, material extracts or in direct cell-material contact

Established testing system (ClicKit-Well) for biological material testing in direct cell-material contact

Conventionally, materials are examined in vitro in cell culture plates, which makes quantitative comparisons between materials (test specimens), e.g., with different geometries, difficult.

The patented in-vitro test system ClicKit-Well was developed for the reliable analysis of surfaces (DE 102018221415 B3). With its surface standardized areas (e.g., 96-well format) on material test specimens, it allows quantitative comparisons between materials and coatings.

Further information incl. a video showing the working principle can be found at the end of the page at “Biomaterial testing 2.0 – standardized, resource-efficient: ClicKit-Well”.

Topic-adressing projects:

• BMWi – WIPANO project (knowledge and technology transfer through patents and standards) with the aim of developing a quantitative direct cytotoxicity test (as a standard test method) for medical devices – cooperation project with Eurofins BioPharma Product Testing Munich GmbH and DIN
• EU-HORIZON cooperation project with the topic “Next Generation BiOactiVe NAnocoatings“ (project acronym: NOVA)
• VDE/VDI BMBF VIP+ cooperation project with the topic “Universally usable in-vitro testing system for standardized, resource-saving implementation of biointerface studies” (project acronym: UniBioface)
• Fraunhofer cooperation: IAP, IFAM, ICT within Simpromat (BMBF-funded; funding code: FKZ 01IO1902): Further development of the in-house developed in-vitro test system with focus on the serial production of individual components and the application in the field of anti-viral testing.
• Fraunhofer Anti-Corona research project with the topic “Effective and robust biocompatibility testing of 3D printed components for the prevention and therapy of COVID-19” (project acronym: BioKomp)

Competencies of the group »Biological Material Analysis«

Biological assessment of materials (in the context of research and development) analogous to DIN EN ISO 10993

• In vitro cytotoxicity tests analogous to ISO 10993 for the assessment of biocompatibility (extract tests, cytotoxicity/adhesion test in direct cell contact)

Method development

• Development of suitable methods and analytical procedures for the biological evaluation of biomaterials/medical products as well as APIs/drugs for industrially transferable screening assays
• Development of novel immuno- and cell biological assays, e.g., for direct application on material surfaces
• Feasibility and validation studies
• Establishment and validation of marker-free/non-destructive analysis methods
• Development of suitable (depending on the problem) test procedures, incl. controls, and preparation of corresponding protocols (SOPs) 

Analytics

• Cell secretome (e.g., ELISA, Luminex)
• Surface markers (e.g., immunofluorescence microscopy, FACS)
• Gene expression (qPCR)
• Enzyme activity determination
• Biochemical assays (e.g., protein, calcium determination)
• MSC characterization according to ISCT criteria (incl. immune potential)

Further competencies of the group »Biological Material Analysis«:

• Regulatory matters for medical devices (Medical device regulation, EU 745/2017)
• Active participation in ISO 10993 (member of DIN committee NA 027-07-12 AA / expert in ISO TC194, contact person: Dr. Susanne Kurz)
• LEAN principles/process optimization 

Selected current projects and research cooperation

• Trilateral transfer project DFG-Fraunhofer with the topic (immune-)biological evaluation “Bioinspired hybrid scaffolds for the bone-tissue-engineering” (Project acronym: SpongeBone)
• VDE/VDI BMBF VIP+ cooperation project with the topic “Universally usable in vitro testing system for standardized, resource-saving implementation of biointerface studies” (Project acronym: UniBioface)
• EU-HORIZON cooperation project with the topic “Next Generation BiOactiVe NAnocoatings“ (Project acronym: NOVA)
• German Korean MOTIE/KIAT cooperation project for the development of a 3D printed medical device containing proteins – cooperation project with L&C Bio and Yonsei University Hospital (both South Korea) and Fraunhofer IZI (Project acronym: ImmuBoost)
• BMWi - WIPANO project (knowledge and technology transfer through patents and standards) with the aim of developing a quantitative direct cytotoxicity test (as a standard test method) for medical devices - cooperation project with Eurofins BioPharma Product Testing Munich GmbH and DIN (Project acronym: ToxQuant)
• German Korean AiF/ZIM cooperation project to establish a marker-free and specific analysis method for the standardized evaluation of non-transparent biomaterials in macrophage activation (cell holography/laser-assisted cell dissection) - Cooperation project with HICS Company and Pixel Inc. (South Korea), LLS ROWIAK LaserLabSolutions GmbH (Germany) (Project acronym: ImplantHoloCut)
• Fraunhofer SME cooperation project (IKTS and IMWS) Process optimization for the evaluation of desensitizing dental care products via the production of an artificial dentin structure based on ceramics (Project acronym: ArtDentin)
• Dissertations within a Fraunhofer Attract project (2016-2021) with the topics »Development of a combined in vitro test scenarios for the evaluation of osteo-immunological properties of medical products«, »Investigation of the influence of experimental conditions on the differentiation of THP-1 cells to polarized macrophages«
• Current dissertation with the topic »Development of a macrophage-based in vitro assay as a predictive model of a foreign body reaction on implantable medical devices«

Publications

Reference | Patent

• Ißleib, C.; Spohn, J.; Kurz, S. (2018). Anordnung zur Durchführung von in vitro Biokompatibilitätsprüfungen. DE102018221415, Espacenet 2020/03/13 [LINK].

Publications

• Ißleib, C.; Kurz, S.; Scholl, S.; Amber, A.; Spohn, J. Plasticity of proinflammatory macrophages depends on their polarization stage during human MSC immunomodulation – an in vitro study using THP-1 and human primary macrophages. Immuno 1(4) (2021) 518-528.
• Blaschke, D.; Pahlow, S.; Fremberg, T.; Weber, K.; Müller, A. D.; Kurz, S.; Spohn, J.; Dhandapani, V.; Rebohle, L.; Skorupa, I.; Schmidt, H. Functionalized silicon substrates with stripe-patterned surface-near electrostatic forces for the self-organized, stable immobilization of biomolecules. Applied Surface Science 545 (2021) 148729.
• Balakin, S.; Yun, Y.-S.; Lee, J.; Kang, E.-H.; Spohn, J.; Yun, I.-S.; Opitz, J.; Cuniberti, G.; Yeo, J.-S. In vitro characterization of osteoblast cells on polyelectrolyte multilayers containing detonation nanodiamonds. Biomed. Mater. 15 (2020) 055026.
• Balakin, S.; Dennison, N. R.; Klemmed, B; Spohn, J.; Cuniberti, G; Romhildt, L.; Opitz, J. Immobilization of Detonation Nanodiamonds on Macroscopic Surfaces. Appl. Sei. 9 (2019) 1064.
• Spohn, J.; Markhoff, J.; Tillmann, J.; Krogull, M.; Pisowocki, P.; Bader, R. Direct Influence of titanium and zirconia particles on the morphology and functionality of mature human osteoclasts. J Biomed Mater Res A. 105 (2017) 36114.

This link (ResearchGate) leads to publications on biological materials analysis by Dr. Juliane Spohn.