The aim of the study was to establish electrical stimulation parameters in order to improve cell growth and viability of human adipose-derived stem cells (hADSC) when compared to non-stimulated cells in vitro. hADSC were exposed to continuous electrical stimulation with 1.7 V AC/20 Hz. After 24, 72 h and 7 days, cell number, cellular surface coverage and cell proliferation were assessed. In addition, cell cycle analysis was carried out after 3 and 7 days. After 24 h, no significant alterations were observed for stimulated cells. At day 3, stimulated cells showed a 4.5-fold increase in cell numbers, a 2.7-fold increase in cellular surface coverage and a significantly increased proliferation. Via cell cycle analysis, a significant increase in the G2/M phase was monitored for stimulated cells. Contrastingly, after 7 days, the non-stimulated group exhibited a 11-fold increase in cell numbers and a 4-fold increase in cellular surface coverage as well as a significant increase in cell proliferation. Moreover, the stimulated cells displayed a shift to the G1 and sub-G1 phase, indicating for metabolic arrest and apoptosis initiation. In accordance, continuous electrical stimulation of hADSC led to a significantly increased cell growth and proliferation after 3 days. However, longer stimulation periods such as 7 days caused an opposite result indicating initiation of apoptosis.

(1) Background: Electrical stimulation is a promising alternative to promote bone fracture healing but with the limitation of tracking the osteogenesis progress in vivo. To overcome this issue, we present an opportunity to combine the electrical stimulation of a commercial titanium implant, which promotes osteogenesis within the fracture, with a real-time readout of the osteogenic progress by impedance sensing. This makes it possible to adjust the electrical stimulation modalities to the individual patient’s fracture healing process. (2) Methods: In detail, osteogenic differentiation of several cell types was monitored under continuous or pulsatile electrical stimulation at 0.7 V AC/20 Hz for at least seven days on a titanium implant by electric cell-substrate impedance sensing (ECIS). For control, chemical induction of osteogenic differentiation was induced. (3) Results: The most significant challenge was to discriminate impedance changes caused by proliferation events from those initiated by osteogenic differentiation. This discrimination was achieved by remodeling the impedance parameter Alpha (α), which increases over time for pulsatile electrically stimulated stem cells. Boosted α-values were accompanied by an increased formation of actin stress fibers and a reduced expression of the focal adhesion kinase in the cell periphery; morphological alterations known to occur during osteogenesis. (4) Conclusions: This work provided the basis for developing an effective fracture therapy device, which can induce osteogenesis on the one hand, and would allow us to monitor the induction process on the other hand.

Metabolic endpoint assays enable rapid screening but provide limited information on treatment kinetics, heterogeneity, and recovery. Here, we established a multistep workflow combining MTS-based metabolic screening with electric cell–substrate impedance sensing (ECIS), a label-free cell-based biosensor, to functionally prioritize complex plant extracts in head and neck squamous cell carcinoma (HNSCC) models. Seventeen soluble extracts were screened at 50 µg/mL for 24 h in four HNSCC cell lines and human adipose-derived stem cells. Selected candidates were characterized by real-time impedance monitoring, multifrequency analysis, and model-derived barrier resistance. Extract 16 showed the most favorable metabolic selectivity profile. ECIS resolved no growth-inhibitory effects, sustained impedance suppression, heterogeneous responses, and transient suppression followed by partial recovery. Recovery was incomplete (CAL-33) or absent (Detroit 562); modeled barrier resistance became resolvable late (FaDu, PE/CA-PJ15) or not at all (CAL-33). Comparison with the PI3Kα inhibitor Inavolisib revealed partially overlapping but non-identical metabolic response profiles and no consistent enhancement by combination treatment. Extract 16 was associated with junctional redistribution, F-actin remodeling, and cell line-dependent PARP cleavage, whereas caspase 3/7 activation could not be detected. Integrating metabolic endpoint screening with multiparametric ECIS monitoring provides greater functional resolution and supports prioritizing complex bioactive samples for chemical and mechanistic investigation.

Metabolic endpoint assays enable rapid screening but provide limited information on treatment kinetics, heterogeneity, and recovery. Here, we established a multistep workflow combining MTS-based metabolic screening with electric cell–substrate impedance sensing (ECIS), a label-free cell-based biosensor, to functionally prioritize complex plant extracts in head and neck squamous cell carcinoma (HNSCC) models. Seventeen soluble extracts were screened at 50 µg/mL for 24 h in four HNSCC cell lines and human adipose-derived stem cells. Selected candidates were subsequently characterized by real-time impedance monitoring, multifrequency analysis, and model-derived barrier resistance. Extract 16 showed the most favorable metabolic selectivity profile. ECIS resolved no growth-inhibitory effects, sustained impedance suppression, heterogeneous responses, and transient suppression followed by partial recovery. Cross-model analysis identified distinct barrier-resistance dynamics, particularly in CAL-33 and Detroit 562. Comparison with the PI3Kα inhibitor Inavolisib revealed partially overlapping but non-identical metabolic response profiles and no consistent enhancement by combination treatment. Extract 16 was further associated with junctional redistribution, F-actin remodeling, and PARP processing, whereas reproducible caspase 3/7 activation was not detected. Thus, integrating metabolic endpoint screening with multiparametric ECIS monitoring provides greater functional resolution and supports the prioritization of complex bioactive samples for subsequent chemical and mechanistic investigation.