Polyblend Nanofibers to Regenerate Gingival Tissue: A Preliminary In Vitro Study

Aim: The regeneration of small periodontal defects has been considered an important divide and challenging issue for dental practitioners. The aim of this preliminary in vitro study was to analyze the effects of polycaprolactone (PCL) nanofibers enriched with hyaluronic acid and vitamin E vs. nude nanofibers on gingival fibroblasts activity, an innovative graft for periodontal soft tissue regeneration purposes. Methods: Nanofibers were produced in PCL (NF) or PCL enriched with hyaluronic acid and vitamin E (NFE) by electrospinning technique. NF and NFE were stereologically and morphologically characterized by scanning electron microscope (SEM), and composition was analyzed by infrared spectroscopy. Human fibroblasts were obtained from one gingival tissue fragment (HGF) and then seeded on NF, NFE, and plastic (CT). Cell adhesion and morphology were evaluated using SEM at 24 h and cell viability after 24, 48, and 72 h by alamarBlue® assay. Gene expression for COL-I, LH2b, TIMP-1, PAX, and VNC was analyzed by real-time RT-PCR in samples run in triplicate and GAPDH was used as housekeeping gene. Slot blot analysis was performed and immunoreactive bands were revealed for MMP-1 and COL-I. YAP and p-YAP were analyzed by Western blot and membranes were reprobed by α-tubulin. Statistical analysis was performed. Results: IR spectrum revealed the presence of PCL in NF and PCL and vitamin E and hyaluronic acid in NFE. At 24 h, HGF adhered on NF and NFE conserving fibroblast like morphology. At 72 h from seeding, statistically significant differences were found in proliferation of HGF cultured on NF compared to NFE. Expression of genes (LH2b, TIMP-1, and MMP-1) and proteins (COL-I) related to collagen turnover revealed a reduction of COL-1 secretion in cells cultured on NF and NFE compared to CT; however, NFE stimulated cross-linked collagen deposition. Mechanosensor genes (PAX, VNC, and YAP) were upregulated in HGF on NF while they were decreased in cells grown on NFE. Conclusion: Preliminary data suggest that PCL-enriched nanofibers could represent a support to induce HGF proliferation, adhesion, collagen cross-linking, and to reduce collagen degradation, therefore favoring collagen deposition in gingival connective tissue

Synthesis of microsponges by spray drying TEMPO-oxidized cellulose nanofibers and characterization for controlled release

A Spray Drying process is presented for one-step production of nanostructured microsponges from TEMPO oxidized cellulose nanofibers (TOCNF). Different formulations were prepared by varying both the nanofibers oxidation degree and the relative ratios of branched polyethyleneimine and citric acid, introduced as crosslinkers. The best reaction conditions were achieved using TOCNF containing the highest content of carboxylic groups (1.5 mmolCOOH/gTOCNF), which produced microparticles with an average diameter below 5 μm, and a good colloidal stability in aqueous dispersion. Physical-chemical characterization was carried out to study the chemical bonds in the microsponges matrix, the swelling degree and drug loading and release. Tetracycline, a broad-spectrum drug with pH-dependent amphoteric feature, was chosen as model molecule. Chemical crosslinking was verified by spectroscopic characterisation, which revealed the formation of amide bonds. The crosslinked microsponges showed a high drug loading efficiency, as opposed to non-cross-linked ones, and a pHresponsivity in terms of swelling and drug release behaviour. Finally, microsponges derived from formulations without citric acid resulted biocompatible, as demonstrated by direct and indirect in vitro tests on gastric cancer cells

TEMPO-Nanocellulose/Ca2+ hydrogels: ibuprofen drug diffusion and In vitro cytocompatibility

Stable hydrogels with tunable rheological properties were prepared by adding Ca2+ ions to aqueous dispersions of 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO)-oxidized and ultra-sonicated cellulose nanofibers (TOUS-CNFs). The gelation occurred by interaction among polyvalent cations and the carboxylic units introduced on TOUS-CNFs during the oxidation process. Both dynamic viscosity values and pseudoplastic rheological behaviour increased by increasing the Ca2+ concentration, confirming the cross-linking action of the bivalent cation. The hydrogels were proved to be suitable controlled release systems by measuring the diffusion coefficient of a drug model (ibuprofen, IB) by high-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy. IB was used both as free molecule and as a 1:1 pre-formed complex with β-cyclodextrin (IB/β-CD), showing in this latter case a lower diffusion coefficient. Finally, the cytocompatibility of the TOUS-CNFs/Ca2+ hydrogels was demonstrated in vitro by indirect and direct tests conducted on a L929 murine fibroblast cell line, achieving a percentage number of viable cells after 7 days higher than 70%