Peculiarities of Cell Seeding on Electroformed Polycaprolactone Scaffolds Modified with Surface-Active Agents Triton X-100 and Polyvinylpyrrolidone.

We compared the capability of human fibroblasts to populate porous polycaprolactone (PCL) scaffolds modified during fabrication with surface-active agents Triton Ð¥-100 (type 1 scaffold) and polyvinylpyrrolidone (type 2 scaffold). The mean fiber diameter in both scaffolds was almost the same: 3.90±2.19 and 2.46±2.15 µ, respectively. Type 1 scaffold had higher surface density and hydrophilicity, when type 2 scaffold was 1.6 times thicker. The cells were seeded on the scaffolds by the dynamic seeding technique and then cultured in Petri dishes with nutrient medium in a humid atmosphere. During 3-day culturing, no cell release from the matrix was noted. DAPI staining proved the presence of cells in both scaffolds. However, in type 1 scaffold the cells populated the whole thickness, while in type 2 scaffold, the cells were present only in the superficial layer. These findings suggest that PCL scaffolds modified with Triton Ð¥-100 or polyvinylpyrrolidone are not cytotoxic, but the structure of the scaffold treated with Triton Ð¥-100 is more favorable for population with cells.

Study of the Osteoindictive Properties of Protein-Modified Polylactide Scaffolds.

Bone marrow mesenchymal stromal cells are multipotent and can differentiate into cells of various tissues, which determines their high importance for clinical application. We performed an in vitro study of the osteogenic potential of mesenchymal stromal cells cultured on intact polylactide scaffolds or scaffolds modified with collagen I or fibrin. Scanning electron microscopy showed that the cells formed osteogenic nodules or osteogenic nodules on both intact and fibrin-modified polylactide scaffolds. Spectrophotometric detection of alkaline phosphatase activity on days 7 and 11 showed that mesenchymal stromal cell grown on intact polylactide scaffolds and on scaffolds modified with collagen type I or fibrin more intensively synthesized alkaline phosphatase than in the control (culture plastic). This dependence increases in the presence of osteogenic differentiation factors in the medium. After long-term culturing (4 weeks), the presence of calcium deposits detected by alizarin red staining confirmed the osteoinductive properties of intact and protein-modified polylactide scaffolds. These findings suggest that polylactide scaffolds and collagen I increase the osteogenic differentiation potential of mesenchymal stromal cells.

Noninvasive penetration of 5 nm hyaluronic acid molecules across the epidermal barrier (in vitro) and its interaction with human skin cells.

OBJECTIVE: Hyaluronic acid represents one of the major components of the extracellular environment. The main challenge remains in the ability to deliver these molecules noninvasively across the skin barrier, which can be overcome by the reduction in size to an extent that allows these molecules to pass across the skin barrier. The aim of this study was to measure the penetration and bioavailability of low molecular weight hyaluronic acid to cross an epidermal barrier model. METHODS: Determining the quantity of hyaluronic acid in the test solutions was carried with method of photocolorimetry analysis. Investigation of the interaction of cells with LMWHA was studied with a confocal microscope. RESULTS: The study showed that LMWHA is able to cross the epidermis. Most effective penetration level is during the first 6 hours reaching 75%, and then the concentration started to decline and reached the equilibrium state within the following 2 hours. Confocal laser microscopy demonstrated different distribution and behavior of these molecules among the keratinocytes and fibroblasts. CONCLUSION: Reducing the size of hyaluronic acid to 5 nm enhance their transport across the epidermal layer. The concentration of hyaluronic acid molecules was higher on the fibroblast surface in comparison to their extracellular environment.

Peculiarities of Cell Seeding on Polylactic Acid-Based Scaffolds Fabricated Using Electrospinning and Solution Blow Spinning Technologies.

We studied the possibility of seeding bone marrow-derived stromal cells onto polylactic acid-based scaffolds fabricated by electrospinning and solution blow spinning technologies. The cells were applied to the scaffolds by dynamic seeding and scaffolds were then cultured in Petri dishes in culture medium for 3 days. Cell migration to the Petri dish surface was noted only for scaffolds fabricated by electrospinning technology, but DAPI staining confirmed the presence of cells in both scaffolds. The mean number of cells in scaffolds fabricated by electrospinning and solution blow spinning was 56±9 and 81±6, respectively. The scaffold fabricated by solution blow spinning was more effectively (p<0.05) colonized by cells due to its more optimal spatial structure.

Effect of Concentration of Collagen Gel on Functional Activity of Bone Marrow Mesenchymal Stromal Cells.

Collagen I gels with protein concentrations of 1, 2, and 3.5 mg/ml were prepared and embedded in a porous polylactide scaffold to reduce their contraction. Concentration of the gel did not affect its degradation. Collagen gels promoted the formation of cell networks. The cells in the collagen gel with a concentration of 1 mg/ml embedded in polylactide scaffold had elongated spindle-like shape, in contrast to flattened cells in collagen gel of the same concentration not embedded in the scaffold. Stabilization of the collagen gel in the polylactide scaffold promoted active synthesis of laminin and fibronectin by cells as soon as on day 5 of culturing in comparison with that in free collagen substrate.