An on-chip wound healing assay fabricated by xurography for evaluation of dermal fibroblast cell migration and wound closure

Abstract

Dermal fibroblast cell migration is a key process in a physiological wound healing. Therefore, the analysis of cell migration is crucial for wound healing research. In this study, lab-on-a-chip technology was used to investigate the effects of basic fibroblast growth factor (bFGF), mitomycin C (MMC), MEK1/2 inhibitor (U0126) and fetal calf serum (FCS) on human dermal fibroblast cell migration. The microdevice was fabricated consisting of microchannels, pneumatic lines and pneumatically-activated actuators by xurographic rapid prototyping. In contrast to current approaches in in vitro wound healing such as scratch assays and silicone inserts in wellplate format, which show high variability and poor reproducibility, the current system aims to automate the wounding procedure at high precision and reproducibility using lab-on-a-chip. Traumatic wounding was simulated on-chip on fibroblast cell monolayers by applying air pressure on the flexible circular membrane actuator. Wound closure was monitored using light microscopy and cell migration was evaluated using image analysis. The pneumatically controlled system generates highly reproducible wound sizes compared to the conventional wound healing assay. As proof-of-principle study wound healing was investigated in the presence of several stimulatory and inhibitory substances and culture including bFGF, MMC, U0126 MEK1/2 inhibitor as well as serum starvation to demonstrate the broad applicability of the proposed miniaturized culture microsystem.

Establishment of a human three-dimensional chip-based chondro-synovial co-culture joint model for reciprocal cross-talk studies in arthritis research

“…They assembled layer by layer by xurography from two-dimensional CAD templates from 500 μm thick transparent silicone sheets (MVQ Silicone) as described previously. 26,27 The basic design principles were adapted from an earlier vascular organoid study 28 , and circular medium reservoirs were added to contain a total medium volume of 300 μl. For height variation, the layer numbers of the circular hydrogel compartment were adjusted in the range of 1-4 mm.…”

Microfluidic and Lab-on-a-Chip Systems for Cutaneous Wound Healing Studies

“…This technology platform created highly automated and reproducible wounds for both methods to show how TNF-α and mitomycin C decreased wound healing speed. A recent follow-up study by Shabestani Monfared et al [54] adapted this approach using PDMS rapid prototyping by xurography to automatically create more wounds with a single pneumatic actuation cycle.…”

“…(D) Effect of growth factor bFGF and inhibitory agents mitomycin C (MMC) and MEK-inhibitor U0126 on dermal fibroblast migration and proliferation dynamics. (ns, non-significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001) Adapted with permission from[54].…”

Microfluidic and Lab-on-a-Chip Systems for Cutaneous Wound Healing Studies

“…This technology platform created highly automated and reproducible wounds for both methods to show how TNF-α and Mitomycin C decreased the wound healing speed. A recent follow-up study by Shabestani Monfared et al [53] adapted this approach using PDMS rapid prototyping by xurography to create more wounds automatically with a single pneumatic actuation cycle.…”

“…D) Effect of growth factor bFGF and inhibitory agents Mitomycin C (MMC) and MEK-inhibitor (U0126) on dermal fibroblast migration and proliferation dynamics. Adapted with permissions from Ref [53]…”