Lars Lehmann

One-Year Clinical and Radiological Results of a Prospective, Investigator-Initiated Trial Examining a Novel, Purely Autologous 3-Dimensional Autologous Chondrocyte Transplantation Product in the Knee:

Background: The 3-dimensional autologous chondrocyte transplantation (ACT3D) comprises isolation of chondrocytes from cartilage biopsies, cultivation to spheroids, and transplantation into the cartilage defect. Objectives: To evaluate the patients’ general health and functionality and to assess the defect repair after ACT3D with spheroids by MRI and MOCART scoring. Methods: Thirty-seven patients with isolated chondral lesions of the knee underwent ACT3D with spheroids through medial arthrotomy. Patient-administered scores were assessed at baseline (day before transplantation), at 6 weeks, and at 3, 6, and 12 months. MRI and MOCART scoring were performed at 3 and 12 months after ACT3D. Results: Patients were diagnosed with full-thickness patellofemoral (n = 16), femoral condylar (n = 18), or both defect types (n = 3), International Cartilage Repair Society (ICRS) grade 3 or 4, with defect sizes between 1.0 and 12.0 cm2. On average, 59.5 spheroids/cm2 in defect size were transplanted. An overall statistically significant improvement from baseline to 12 months was observed for all assessment scores (Lysholm, International Knee Documentation Committee [IKDC], SF-36, Tegner) combined with a significant reduction in the visual analog scale (VAS) for pain and an advanced defect filling. Subgroup analyses revealed a positive clinical outcome independent on defect size, defect locations, spheroid dosage, age, duration of symptoms, and severity of complaints at baseline. Seven patients experienced in total 8 adverse events, of which knee joint effusion and blocking were assessed as possibly or probably related to ACT3D. Conclusions: The patient-administered assessment scores along with the fast defect filling with ACT3D using spheroids demonstrated an increase in activity level and quality of life after a 1-year follow-up.

Live-cell assays to identify regulators of ER-to-Golgi trafficking.

We applied fluorescence microscopy‐based quantitative assays to living cells to identify regulators of endoplasmic reticulum (ER)‐to‐Golgi trafficking and/or Golgi complex maintenance. We first validated an automated procedure to identify factors which influence Golgi‐to‐ER relocalization of GalT‐CFP (β1,4‐galactosyltransferase I‐cyan fluorescent protein) after brefeldin A (BFA) addition and/or wash‐out. We then tested 14 proteins that localize to the ER and/or Golgi complex when overexpressed for a role in ER‐to‐Golgi trafficking. Nine of them interfered with the rate of BFA‐induced redistribution of GalT‐CFP from the Golgi complex to the ER, six of them interfered with GalT‐CFP redistribution from the ER to a juxtanuclear region (i.e. the Golgi complex) after BFA wash‐out and six of them were positive effectors in both assays. Notably, our live‐cell approach captures regulator function in ER‐to‐Golgi trafficking, which was missed in previous fixed cell assays, as well as assigns putative roles for other less characterized proteins. Moreover, we show that our assays can be extended to RNAi and chemical screens.

Scintigraphic evaluation of rhBMP-2-biocoated implants reveals no ectopic bone formation

The main objectives of the study described below were of two-fold nature: (1) to examine if rhBMP-2-biocoated implants in a pig model could lead to ectopic bone formation and (2) if quantitative and/or qualitative differences could be found between adhesively and covalently bonded BMP II using the scintigraphic method. In order to examine these central questions, 26 Göttingen minipigs were allocated to three groups with a control group (n=7) and two study groups (n=9 each) receiving one of three implant types: (a) chromosulfuric acid treated titanium surface as control, (b) non-covalently bonded BMP-2, and (c) covalently bonded and immobilized rhBMP-2. Each animal received four barbell-shaped implants, one in the proximal and distal metaphysis of each femur. The scintigraphic analyses were conducted after four, eight, and 12 weeks postoperatively. The visual (qualitative) analysis failed to show ectopic bone formation in any of the three groups. The statistical analysis of the relative values for bone formation yielded no significant differences between the groups, although the limitation in the applied methods do not enable one to draw conclusions regarding the histomophometric results.