'Pre-prosthetic use of poly(lactic-co-glycolic acid) membranes treated with oxygen plasma and TiO2 nanocomposite particles for guided bone regeneration processes'.

OBJECTIVES: Guided bone regeneration (GBR) processes are frequently necessary to achieve appropriate substrates before the restoration of edentulous areas. This study aimed to evaluate the bone regeneration reliability of a new poly-lactic-co-glycolic acid (PLGA) membrane after treatment with oxygen plasma (PO2) and titanium dioxide (TiO2) composite nanoparticles. METHODS: Circumferential bone defects (diameter: 10mm; depth: 3mm) were created on the parietal bones of eight experimentation rabbits and were randomly covered with control membranes (Group 1: PLGA) or experimental membranes (Group 2: PLGA/PO2/TiO2). The animals were euthanized two months afterwards, and a morphologic study was then performed under microscope using ROI (region of interest) colour analysis. Percentage of new bone formation, length of mineralised bone formed in the grown defects, concentration of osteoclasts, and intensity of osteosynthetic activity were assessed. Comparisons among the groups and with the original bone tissue were made using the Kruskal-Wallis test. The level of significance was set in advance at a=0.05. RESULTS: The experimental group recorded higher values for new bone formation, mineralised bone length, and osteoclast concentration; this group also registered the highest osteosynthetic activity. Bone layers in advanced formation stages and low proportions of immature tissue were observed in the study group. CONCLUSIONS: The functionalised membranes showed the best efficacy for bone regeneration. CLINICAL SIGNIFICANCE: The addition of TiO2 nanoparticles onto PLGA/PO2 membranes for GBR processes may be a promising technique to restore bone dimensions and anatomic contours as a prerequisite to well-supported and natural-appearing prosthetic rehabilitations.

Osteoconductive potential of barrier nanoSiO2 PLGA membranes functionalized by plasma enhanced chemical vapour deposition.

The possibility of tailoring membrane surfaces with osteoconductive potential, in particular in biodegradable devices, to create modified biomaterials that stimulate osteoblast response should make them more suitable for clinical use, hopefully enhancing bone regeneration. Bioactive inorganic materials, such as silica, have been suggested to improve the bioactivity of synthetic biopolymers. An in vitro study on HOB human osteoblasts was performed to assess biocompatibility and bioactivity of SiO2 functionalized poly(lactide-co-glycolide) (PLGA) membranes, prior to clinical use. A 15 nm SiO2 layer was deposited by plasma enhanced chemical vapour deposition (PECVD), onto a resorbable PLGA membrane. Samples were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, scanning electron microscopy, and infrared spectroscopy (FT-IR). HOB cells were seeded on sterilized test surfaces where cell morphology, spreading, actin cytoskeletal organization, and focal adhesion expression were assessed. As proved by the FT-IR analysis of samples, the deposition by PECVD of the SiO2 onto the PLGA membrane did not alter the composition and other characteristics of the organic membrane. A temporal and spatial reorganization of cytoskeleton and focal adhesions and morphological changes in response to SiO2 nanolayer were identified in our model. The novedous SiO2 deposition method is compatible with the standard sterilization protocols and reveals as a valuable tool to increase bioactivity of resorbable PLGA membranes.

In vivo comparative model of oxygen plasma and nanocomposite particles on PLGA membranes for guided bone regeneration processes to be applied in pre-prosthetic surgery: a pilot study.

OBJECTIVES: To evaluate the bone regeneration potential of a new membrane fabricated with polyglycolide acid (PLGA) after being treated with oxygen plasma (PO2), and/or being functionalized with silicon dioxide (SiO2) or titanium dioxide (TiO2) nanoparticles. METHODS: Bone defects (5 mm 3 mm) were produced on the top of 3 experimentation rabbits' skulls and were covered with variously modified PLGA scaffolds. After the animals were sacrificed, neoformed bone (%), mineralized bone (mm), bone resorption (%), osteoclasts/mm2, and intensity of osteosynthetic activity, were assessed under microscope. RESULTS: The following groups were formed depending on the type of membrane: PLGA (control); PLGA/PO2; PLGA/SiO2; PLGA/TiO2; PLGA/PO2/SiO2; and PLGA/PO2/TiO2. The histological sections showed bone layers in advanced stages of formation. The highest percentages of neoformed bone corresponded to PLGA/PO2/SiO2 membranes (59.07%; p = 0.31) followed by PLGA/PO2 barriers (50.27%). The controls showed the lowest mineralization (13.89 mm; p = 0.24). PLGA/TiO2 scaffolds exhibited the least bone resorption (4.45%; p = 0.77) and osteoclasts/ mm2 (1.58; p = 0.86). PLGA/SiO2 and PLGA/TiO2 membranes stimulated the maximum osteosynthetic activity. CONCLUSIONS: The treatment of PLGA barriers with PO2 increased bone regeneration in rabbits. When comparing the effect of PO2/SiO2 and PO2/TiO2, higher percentages of neoformed bone were encountered after silicon-dioxide coating. CLINICAL SIGNIFICANCE: The incorporation of SiO2 nanoparticles onto PO2-treated PLGA membranes was the most promising technique out of those investigated to promote bone formation in rabbits. The addition of SiO2 or TiO2 layers to PLGA substrates may stimulate the osteosynthetic activity, which might be useful to restore bone dimensions in preparation for naturally appearing dental prostheses.

Osteoblasts Interaction with PLGA Membranes Functionalized with Titanium Film Nanolayer by PECVD. In vitro Assessment of Surface Influence on Cell Adhesion during Initial Cell to Material Interaction.

New biomaterials for Guided Bone Regeneration (GBR), both resorbable and non-resorbable, are being developed to stimulate bone tissue formation. Thus, the in vitro study of cell behavior towards material surface properties turns a prerequisite to assess both biocompatibility and bioactivity of any material intended to be used for clinical purposes. For this purpose, we have developed in vitro studies on normal human osteoblasts (HOB®) HOB® osteoblasts grown on a resorbable Poly (lactide-co-glycolide) (PLGA) membrane foil functionalized by a very thin film (around 15 nm) of TiO2 (i.e., TiO2/PLGA membranes), designed to be used as barrier membrane. To avoid any alteration of the membranes, the titanium films were deposited at room temperature in one step by plasma enhanced chemical vapour deposition. Characterization of the functionalized membranes proved that the thin titanium layer completely covers the PLGA foils that remains practically unmodified in their interior after the deposition process and stands the standard sterilization protocols. Both morphological changes and cytoskeletal reorganization, together with the focal adhesion development observed in HOB osteoblasts, significantly related to TiO2 treated PLGA in which the Ti deposition method described has revealed to be a valuable tool to increase bioactivity of PLGA membranes, by combining cell nanotopography cues with the incorporation of bioactive factors.

Light induced hydrophilicity and osteoblast adhesion promotion on amorphous TiO2.

We have studied the effect of the UV induced superhydrophilic wetting of TiO(2) thin films on the osteoblasts cell adhesion and cytoskeletal organization on its surface. To assess any effect of the photo-catalytic removal of adventitious carbon as a factor for the enhancement of the osteoblast development, 100 nm amorphous TiO(2) thin layers were deposited on polyethylene terephthalate (PET), a substrate well known for its poor adhesion and limited wettability and biocompatibility. The TiO(2) /PET materials were characterized by X-ray photoelectron spectroscopy, and atomic force microscopy and their wetting behavior under light illumination studied by the sessile drop method. The amorphous TiO(2) thin films showed a very poor photo-catalytic activity even if becoming superhydrophilic after illumination. The illuminated samples recovered partially its initial hydrophobic state only after their storage in the dark for more than 20 days. Osteoblasts (HOB) were seeded both on bare PET and on TiO(2) /PET samples immediately after illumination and also after four weeks storage in darkness. Cell attachment was much more efficient on the immediately illuminated TiO(2)/PET samples, with development of focal adhesions and cell traction forces. Although we cannot completely discard some photo-catalytic carbon removal as a factor contributing to this cell enhanced attachment, our photodegradation experiments on amorphous TiO(2) are conclusive to dismiss this effect as the major cause for this behavior.

Roughness assessment and wetting behavior of fluorocarbon surfaces.

The wetting behavior of fluorocarbon materials has been studied with the aim of assessing the influence of the surface chemical composition and surface roughness on the water advancing and receding contact angles. Diamond like carbon and two fluorocarbon materials with different fluorine content have been prepared by plasma enhanced chemical vapor deposition and characterized by X-ray photoemission, Raman and FT-IR spectroscopies. Very rough surfaces have been obtained by deposition of thin films of these materials on polymer substrates previously subjected to plasma etching to increase their roughness. A direct correlation has been found between roughness and water contact angles while a superhydrophobic behavior (i.e., water contact angles higher than 150° and relatively low adhesion energy) was found for the films with the highest fluorine content deposited on very rough substrates. A critical evaluation of the methods currently used to assess the roughness of these surfaces by atomic force microscopy (AFM) has evidenced that calculated RMS roughness values and actual surface areas are quite dependent on both the scale of observation and image resolution. A critical discussion is carried out about the application of the Wenzel model to account for the wetting behavior of this type of surfaces.