Microfabricated Gaps Reveal the Effect of Geometrical Control in Wound Healing

The geometry (size and shape) of gaps is a key determinant in controlling gap closure during wound healing. However, conventional methods for creating gaps result in un‐defined geometries and poorly characterized conditions (cell death factors and cell debris), which can influence the gap closure process. To overcome these limitations, a novel method to create well‐defined geometrical gaps is developed. First, smooth muscle cells (SMCs) are seeded in variously shaped micro‐containers made out of hyaluronic acid hydrogels. Cell proliferation and cell tension induce fibrous collagen production by SMCs predominantly around the edges of the micro‐containers. Upon removal of SMCs, the selectively deposited collagen results in micro‐containers with cell‐adhesive regions along the edges and walls. Fibroblasts are seeded in these micro‐containers, and upon attaching and spreading, they naturally form gaps with different geometries. The rapid proliferation of fibroblasts from the edge results in filling and closure of the gaps. It is demonstrated that gap closure rate as well as closure mechanism is strongly influenced by geometrical features, which points to an important role for cellular tension and cell proliferation in gap closure

Microfabricated Gaps Reveal the Effect of Geometrical Control in Wound Healing

The geometry (size and shape) of gaps is a key determinant in controlling gap closure during wound healing. However, conventional methods for creating gaps result in un‐defined geometries and poorly characterized conditions (cell death factors and cell debris), which can influence the gap closure process. To overcome these limitations, a novel method to create well‐defined geometrical gaps is developed. First, smooth muscle cells (SMCs) are seeded in variously shaped micro‐containers made out of hyaluronic acid hydrogels. Cell proliferation and cell tension induce fibrous collagen production by SMCs predominantly around the edges of the micro‐containers. Upon removal of SMCs, the selectively deposited collagen results in micro‐containers with cell‐adhesive regions along the edges and walls. Fibroblasts are seeded in these micro‐containers, and upon attaching and spreading, they naturally form gaps with different geometries. The rapid proliferation of fibroblasts from the edge results in filling and closure of the gaps. It is demonstrated that gap closure rate as well as closure mechanism is strongly influenced by geometrical features, which points to an important role for cellular tension and cell proliferation in gap closure

Hot embossing for fabrication of a microfluidic 3D cell culture

Clinically relevant studies of cell function in vitro require a physiologically-representative microenvironment possessing aspects such as a 3D extracellular matrix (ECM) and controlled biochemical and biophysical parameters. A polydimethylsiloxane (PDMS) microfluidic system with a 3D collagen gel has previously served for analysis of factors inducing different responses of cells in a 3D microenvironment under controlled biochemical and biophysical parameters. In the present study, applying the known commercially-viable manufacturing methods to a cyclic olefin copolymer (COC) material resulted in a microfluidic device with enhanced 3D gel capabilities, controlled surface properties, and improved potential to serve high-volume applications. Hot embossing and roller lamination molded and sealed the microfluidic device. A combination of oxygen plasma and thermal treatments enhanced the sealing, ensured proper placement of the 3D gel, and created controlled and stable surface properties within the device. Culture of cells in the new device indicated no adverse effects of the COC material or processing as compared to previous PDMS devices. The results demonstrate a methodology to transition microfludic devices for 3D cell culture from scientific research to high-volume applications with broad clinical impact.National Cancer Institute (U.S.) (award R21CA140096)Charles Stark Draper Laboratory (IR&D Grant

Dysmetabolic circulating tumor cells are prognostic in metastatic breast cancer

Circulating tumor cells (CTCs) belong to a heterogeneous pool of rare cells, and a unequivocal phenotypic definition of CTC is lacking. Here, we present a definition of metabolically-altered CTC (MBA-CTCs) as CD45-negative cells with an increased extracellular acidification rate, detected with a single-cell droplet microfluidic technique. We tested the prognostic value of MBA-CTCs in 31 metastatic breast cancer patients before starting a new systemic therapy (T0) and 3\u20134 weeks after (T1), comparing results with a parallel FDA-approved CellSearch (CS) approach. An increased level of MBA-CTCs was associated with: I) a shorter median PFS pre-therapy (123 days vs. 306; p < 0.0001) and during therapy (139 vs. 266 days; p = 0.0009); ii) a worse OS pre-therapy (p = 0.0003, 82% survival vs. 20%) and during therapy (p = 0.0301, 67% survival vs. 38%); iii) good agreement with therapy response (kappa = 0.685). The trend of MBA-CTCs over time (combining data at T0 and T1) added information with respect to separate evaluation of T0 and T1. The combined results of the two assays (MBA and CS) increased stratification accuracy, while correlation between MBA and CS was not significant, suggesting that the two assays are detecting different CTC subsets. In conclusion, this study suggests that MBA allows detection of both EpCAM-negative and EpCAM-positive, viable and label-free CTCs, which provide clinical information apparently equivalent and complementary to CS. A further validation of proposed method and cut-offs is needed in a larger, separate study

3D microniches reveal the importance of cell size and shape

Contains fulltext : 187738.pdf (publisher's version ) (Open Access)12 p

Salud y seguridad laboral del obrero de la construcción: La ideología empresarial

No pose

Microfabricated Gaps Reveal the Effect of Geometrical Control in Wound Healing

Contains fulltext : 231314.pdf (Publisher’s version ) (Open Access)19 oktober 20208 p