Case Study: microCT, HR-pQCT, qBEI and Raman micro-spectroscopy at UKE, Hamburg

The Institute of Osteology and Biomechanics, UKE uses XamFlow to analyse images from various modalities using a multitide of workflows, of which some are presented below.

“I’m really impressed!” – Dr. Dr. Felix von Brackel, Institute of Osteology and Biomechanics, UKE

Micro-computed tomography analysis of bone

The bone mass and microstructure for example of the femoral neck are evaluated using micro-computed tomography, and parameters such as the following are quantified in XamFlow:

  • Cortical porosity, Ct.Po

  • Cortical thickness, Ct.Th

  • Cortical tissue mineral density, Ct.TMD

  • Cortical pore diameter, Ct.Po.Dm

  • Cortical tissue mineral density, Ct.TMD

  • Bone volume per total volume, BV/TV

  • Trabecular thickness, Tb.Th

  • Trabecular separation, Tb.Sp

  • Trabecular bone mineral density Tb.BMD

Subchondral cyst analysis

For a specialized analysis of subchondral cysts in femoral heads, where multiple samples of femoral heads were measured with SCANCO XtremeCT II (17 GB per image, 6 images analysed initially), a four stage segmentation workflow was implemented:

  1. Bone mask and hull

  2. Cartilage mask and loss area

  3. Subchondral area mask

  4. Cysts

A curvelet filter was used to identify the subchondral area.

../_images/casestudy_femur_workflow.png
../_images/casestudy_femur_compare_blend.png

Quantification of vessel calcification

HR-pQCT image stacks are batch loaded and processed via a defined workflow to detect and quantify LLAC (lower leg arterial calcification) in the soft tissue surrounding the bones. Mineralized areas (excluding the bones) are evaluated with respect to density and volume.

Assessment of lacunar and porosity indices using μCT

Vertebral, trabecular and cortical lacunar morphology in the femur regions are segmented and analysed automatically. Lacunar shape parameters and the cortical vascular canal system are also evaluated. Image segmentation, analysis and visualization are performed in XamFlow.

  • Lacunar volume, Lc.V

  • Lacunar number, Lc.N or N.Lc

  • Lacunar number density, N.Lc/BV

  • Lacunar volume density, Lc.V/BV

  • Individual mean lacunar volume, Lc.V/N.Lc

  • Lacunar sphericity, Lc.Sph or Lc.Sr

  • Lacunar stretch, Lc.St

  • Lacunar oblateness, Lc.O or Lc.Ob

  • Lacunar orientation Lc.Θ

  • Canal porosity, Ca.V/Ca.TV

  • Canal number density, N.Ca/Ct.TV

  • Canal diameter, Ca.Dm

  • Canal separation, Ca.Sp

../_images/casestudy_uke_osteocyte_vascular.png

Quantitative backscattered electron imaging (qBEI)

The mineral distribution is assessed dependent of the distance to the osteocyte lacunar wall, by segmenting osteocyte lacunae in XamFlow.

Raman micro-spectroscopy

Spectral processing of high-resolution (HR) Raman maps of mouse tibiae is performed using XamFlow. Steps like cosmic ray removal and the subtraction of the fluorescent background are automatically applied to all locations in the map. The area of specific bands (like the Amide III band and MMA) are integrated and divided to calculate ratios (e.g. the MMA/Amide III ratio). Spectral peaks are calculated to acquire additional ratios (e.g. the MMA/mineral ratio).

References

Investigation of osseointegration in dental implants influenced by loading and drilling protocols A M Batouche, B Busse, J Krug, G Lotze, F Roosen-Runge, S Galli Osteologie, 2025, A43 https://doi.org/10.1055/s-0045-1804975

Fracture characteristics of human cortical bone influenced by the duration of in vitro glycation Mei-Chun Lin, Praveer Sihota, Sofie Dragoun Kolibová, Imke A.K. Fiedler, Johannes Krug, Eva M. Wölfel, Manuela Moritz, Maria Riedner, Benjamin Ondruschka, Mustafa Citak, Felix Klebig, Felix N. von Brackel, Mahan Qwamizadeh, Katharina Jähn-Rickert and Björn Busse JBMR Plus, ziae151, November 2024 https://doi.org/10.1093/jbmrpl/ziae151

Bone-seeking tumor cells alter bone material quality parameters on the nanoscale in mice Johannes Krug, Christine Plumeyer, Anton Davydok, Sofie Dragoun Kolibova, Nico Fischer, Xuan-Thanh Le-Phuoc, Martina Rauner, Praveer Sihota, Michaela Schweizer, Björn Busse, Imke A.K. Fiedler, Katharina Jähn-Rickert Biomaterials Advances, 2024, 214060, ISSN 2772-9508 https://doi.org/10.1016/j.bioadv.2024.214060

High-fiber diet reduces bone formation but does not affect bone microarchitecture in type 2 diabetes individuals Malak Faraj, Giulia Leanza, Johannes Krug, Francesca Cannata, Viola Viola, Biagio Zampogna, Fabrizio Russo, Giuseppe Banfi, Giovanni Lombardi, Gianluca Vadalà, Laura Mangiavini, Rocco Papalia, Vincenzo Denaro, Björn Busse, Nicola Napoli JBMR Plus, Volume 8, Issue 10, October 2024, ziae111 https://doi.org/10.1093/jbmrpl/ziae111

Sex-Specific Association Patterns of Bone Microstructure and Lower Leg Arterial Calcification Mikolaj Bartosik, Alexander Simon, Björn Busse, Florian Barvencik, Michael Amling, Ralf Oheim & Felix N. von Brackel Microstructure and Lower Leg Arterial Calcification. Calcif Tissue Int (2024). https://doi.org/10.1007/s00223-024-01299-w

Utility and Limitations of TALLYHO/JngJ as a Model for Type 2 Diabetes–Induced Bone Disease Lejla Emini, Juliane Salbach-Hirsch, Johannes Krug, Katharina Jähn-Rickert, Björn Busse, Martina Rauner, Lorenz C. Hofbauer JBMR Plus, e10843, 2023 https://doi.org/10.1002/jbm4.10843

When Cortical Bone Matrix Properties Are Indiscernible between Elderly Men with and without Type 2 Diabetes, Fracture Resistance Follows Suit. Wölfel, E.M., Bartsch, B., Koldehoff, J., Fiedler, I.A.K., Dragoun-Kolibova, S., Schmidt, F.N., Krug, J., Lin, M.-C., Püschel, K., Ondruschka, B., Zimmermann, E.A., Jelitto, H., Schneider, G., Gludovatz, B. and Busse, B. JBMR Plus, e10839, 2023 https://doi.org/10.1002/jbm4.10839

Human tibial cortical bone with high porosity in type 2 diabetes mellitus is accompanied by distinctive bone material properties Eva M. Wölfel, Imke A.K. Fiedler, Sofie Dragoun Kolibova, Johannes Krug, Mei-Chun Lin, Bashar Yazigi, Anna K. Siebels, Herbert Mushumba, Birgit Wulff, Benjamin Ondruschka, Klaus Püschel, Claus C. Glüer, Katharina Jähn-Rickert, Björn Busse Bone, Volume 165, 2022, 116546, ISSN 8756-3282 https://doi.org/10.1016/j.bone.2022.116546

Influence of X-rays and gamma-rays on the mechanical performance of human bone factoring out intraindividual bone structure and composition indices. Schmidt, Felix N; Hahn, Michael; Stockhausen, Kilian E; Rolvien, Tim; Schmidt, Constantin; Knopp, Tobias; Schulze, Christian; Püschel, Klaus; Amling, Michael; Busse, Björn. Materials today. Bio, Vol. 13, 01.2022, p. 100169. https://doi.org/10.1016/j.mtbio.2021.100169

Superior Bone Microarchitecture in Anatomic Versus Nonanatomic Fibular Drill Tunnels for Reconstruction of the Posterolateral Corner of the Knee Julian Stürznickel, Felix N. Schmidt, Conradin Schweizer, Herbert Mushumba, Matthias Krause, Klaus Püschel, and Tim Rolvien, Orthopaedic journal of sports medicine vol. 10,9 23259671221126475. 28 Sep. 2022 https://doi.org/10.1177/23259671221126475

Bone healing and reactivation of remodeling under asfotase alfa therapy in adult patients with pediatric-onset hypophosphatasia. Stürznickel, J., Schmidt, F. N., von Vopelius, E., Delsmann, M. M., Schmidt, C., Jandl, N. M., Oheim, R., & Barvencik, F. Bone, 143, 2021, 115794. https://doi.org/10.1016/j.bone.2020.115794

Subchondral cyst analysis:

Rossinelli, D. (2019).

Three-dimensional analysis of subchondral cysts in hip osteoarthritis: an ex vivo HR-pQCT study. Chiba, K., Burghardt, A. J., Osaki, M., & Majumdar, S. Bone, 66, 140–145. (2014) https://doi.org/10.1016/j.bone.2014.06.001