Clinical perspective.

<p>To determine the effect of mechanical stress on cell survival during suturing the metabolic activity of cell-filled sutures was determined before and after suturing in an <i>ex vivo</i> model using human skin, where no significant difference were observed (A). The potential to produce storable cell-filled sutures was evaluated. Therefore, the metabolic activity of ASCs in the suture was determined after freezing relative to unfrozen sutures. After freezing, 78% of the metabolic activity was preserved (B). p<0.001.</p

Cytokine release from ASC-filled sutures.

<p>A cytokine array detected a multitude of different cytokines released from ASC-filled sutures (A). VEGF was constantly released to cell culture medium for at least 16 days (B). VEGF and SDF-1α were detected in protein extracts of ASC-filled sutures 16 days after seeding (C).</p

Cellular distribution and attachment in the suture.

<p>ASCs were filled into biodegradable sutures. Laser scanning microscopy (LSM, A) shows attached cells (DAPI, blue) along the suture surface interacting with each other (phalloidin, green) and the suture itself (DAPI, blue/phalloidin, green). The suture material is autofluorescent. As observed by scanning electron microscopy (SEM, B), cells were distributed throughout the surface (left) and the inner filaments (middle, right) of the suture. Scale bars represent 50 (A and B left), 500 (B, middle), and 100 μm (B, right).</p

Biomechanical properties of ASC-filled sutures.

<p>A force/expansion profile (A) was recorded to evaluate stiffness (B), maximum force (C), and the elastic limit (D) of ASC-filled sutures. Results show that stiffness and maximum force were significantly reduced when compared to cell-free sutures (control), but the elastic limit remained similar. p<0.001.</p

Biofunctionality of ASC-filled sutures.

<p>Fresh media (control) and media conditioned with cell-filled sutures (conditioned) were tested in an <i>in vitro</i> wound healing assay (scratch assay) with ASCs. After 30 hours, a significant reduction in the open wound was observed when conditioned media was applied. Scale bars represent 500 μm. p<0.05.</p

Cell characterization.

<p>After isolation, cells adhered to tissue culture plastic showing a fibroblast-like morphology (A, DAPI (blue)/phalloidin (red) staining) and multipotency (B–D). Chondrogenic (B), adipogenic (C) and osteogenic (D) differentiation potential was confirmed by Alcian blue, Oil red O and von Kossa staining, respectively. Scale bars represent 100 μm in A, C, D and 1 mm in B.</p

Cell viability and proliferation in the suture.

<p>The metabolic activity of ASCs in the suture was determined by MTT assay. Metabolically active cells (dark precipitate) were observed on the surface (overview) and in the inner cavity (cross section) of the suture at day 1 and 8 after seeding. Moreover, a significant increase of metabolic activity was observed from day 1 to 8. Scale bar represents 2 mm. p<0.05.</p

Uncertainties and controversies in axillary management of patients with breast cancer

The aims of this Oncoplastic Breast Consortium and European Breast Cancer Research Association of Surgical Trialists initiative were to identify uncertainties and controversies in axillary management of early breast cancer and to recommend appropriate strategies to address them. By use of Delphi methods, 15 questions were prioritized by more than 250 breast surgeons, patient advocates and radiation oncologists from 60 countries. Subsequently, a global virtual consensus panel considered available data, ongoing studies and resource utilization. It agreed that research should no longer be prioritized for standardization of axillary imaging, de-escalation of axillary surgery in node-positive cancer and risk evaluation of modern surgery and radiotherapy. Instead, expert consensus recommendations for clinical practice should be based on current evidence and updated once results from ongoing studies become available. Research on de-escalation of radiotherapy and identification of the most relevant endpoints in axillary management should encompass a meta-analysis to identify knowledge gaps, followed by a Delphi process to prioritize and a consensus conference to refine recommendations for specific trial designs. Finally, treatment of residual nodal disease after surgery was recommended to be assessed in a prospective register.</p