Biology subjects

Gorgels, A.

Publications and source records attributed to Gorgels, A..

3 recordsLinked to original sources

bioRxiv2026

Single-cell characterization of human periodontal ligament (PDL) cells reveals uniformly mesenchymal populations in matched maxilla and mandible pairs from four donors

The periodontal ligament (PDL) is a mechanically active connective tissue whose fibroblasts and resident progenitor cells drive tooth support, remodeling, and regeneration. Although PDL cell populations are known to be heterogeneous, the composition of expanded primary PDL cultures and the extent to which anatomical origin shapes their transcriptome remain incompletely defined. Here we applied single-cell RNA sequencing to cultured primary PDL cells isolated from matched upper (maxilla) and lower (mandible) jaw sites of four donors, yielding 43,423 cells that were integrated across donors and analyzed by unsupervised clustering. The expanded cultures were uniformly mesenchymal and fibroblastic, essentially devoid of endothelial and immune signatures, and resolved into ten functionally distinct fibroblast substates rather than discrete cell types, including proliferating, collagen- and extracellular-matrix-high, contractile, and progenitor-like populations. Donor identity was the dominant source of transcriptional variation, with several substates restricted to individual donors. A donor-controlled comparison of maxilla versus mandible revealed only a small set of differentially expressed genes, headed by craniofacial positional-identity transcription factors (PITX1 higher in the mandible; BARX1, ALX1, and NKX6-1 higher in the maxilla), indicating that cultured PDL cells retain embryonic positional memory. In contrast, curated wound-healing gene modules did not differ significantly between jaws, and the transcriptional data therefore did not support the clinical impression of faster healing in the maxilla, suggesting that the healing asymmetry more likely reflects vascular, mechanical, and inflammatory signaling factors acting beyond the steady-state transcriptome. These findings characterize the cellular composition of expanded PDL cultures and identify positional identity as the principal jaw-dependent transcriptional feature.

genomics↗

bioRxiv2025

Cell-based calcification assays - magnesium overrules calcium phosphate saturation in overall outcome

Soft tissue calcification, a common cause of cardiovascular mortality in conditions like chronic kidney disease (CKD), is influenced by magnesium (Mg). However, the exact mechanism, whether extracellular chemistry-mediated or cell-mediated, remains unclear. Here we focused on the extracellular milieu. Using fluorescence-labelled fetuin-A and a live imaging platform, we found that Mg reduced spontaneous mineral precipitation by stabilizing calcium and phosphate as colloidal protein-mineral particles (CPP) in calcification media containing up to 10 mM calcium and phosphate. Addition of 1.25-5 mM Mg progressively stabilized the protein-mineral particles in the fluid phase, increasing calcification. Dynamic light scattering revealed smaller and more numerous CPP at up to 4 mM Mg. At 10 mM Mg, no calcification occurred even with 10 mM added calcium and phosphate. This suggested a critical Mg concentration range in supersaturated calcification media, where Mg paradoxically enhanced calcification by stabilizing mineral precursors and increasing their availability. These findings demonstrate Mgs dual role: an inhibitor of protein-mineral complex aggregation at high concentrations and a facilitator of mineral availability from colloidal protein-mineral phases at concentrations up to about 5 mM Mg. This highlights the role of Mg in providing highly dynamic, mineral-rich, stable protein-mineral complexes driving matrix calcification, yet preventing mineral precipitation.

pathology↗

bioRxiv2025

Miniaturized Device for Assessing Calcification Propensity of Biohybrid Implants Under Continuous Flow

Biohybrid implants are a promising development for cardiovascular disease treatment but suffer from problems like thrombogenesis and calcification. However, testing and validating biohybrid implants can be difficult and expensive due to material handling, fabrication methods and specialty medium components. The devices used to test potential samples can be large and expensive, requiring significant amounts of cell culture medium to operate. Additional, conventional static cell culture conditions do not accurately represent the vascular environment as shear and mechanical forces play key roles in the development of calcification. To address these challenges, a miniaturized, dual-channel flow chamber was designed and validated that allowed for real-time visualization of biohybrid calcification in a physiological environment. Computational Fluid Dynamics simulations were performed to determine the flow characteristics that generated physiological shear stress homogeneously across the sample surface. Micro particle tracking velocimetry measurements validated the simulated shear stresses near the sample surface. Two implant materials used for biohybrid construction, bovine pericardium and polycarbonate urethane, were inserted in the device and exposed to a flowing calcification medium for 14 days. Fluorescent fetuin-A was introduced into the calcification medium for real-time calcification monitoring. The two materials were compared with matched samples calcified in a large fatigue tester for 14 days. Our results showed similar material calcification for bovine pericardium and no calcification for polycarbonate urethane in the large fatigue tester and in our newly developed device. Biohybrid textile-reinforced fibrin-based scaffold populated with vascular smooth muscle cells started to calcify over 7 days in calcification medium. We conclude that this platform will provide novel insights into the origin and progression of pathological calcification and its potentially harmful health effects, which can occur as a result of tissue or metabolic abnormalities, disease, or implantation of certain biomaterials, by providing the ability to monitor the progression of calcification in biohybrid implants in real time, while also minimizing the cost and size of samples and reagents required for testing.

bioengineering↗