Engineering Interfacial Environment of Epigallocatechin Gallate Coated Titanium for Next-Generation Bioactive Dental Implant Components.

In view of endowing the surface of abutments, a component of titanium dental implant systems, with antioxidant and antimicrobial properties, a surface layer coated with epigallocatechin gallate (EGCg), a polyphenol belonging to the class of flavonoids, was built on titanium samples. To modulate interfacial properties, EGCg was linked either directly to the surface, or after populating the surface with terminally linked polyethyleneglycol (PEG) chains, Mw ~1600 Da. The underlying assumption is that fouling-resistant, highly hydrated PEG chains could reduce non-specific bioadhesion and magnify intrinsic EGCg properties. Treated surfaces were investigated by a panel of surface/interfacial sensitive techniques, to provide chemico-physical characterization of the surface layer and its interfacial environment. Results show: (i) successful EGCg coupling for both approaches; (ii) that both approaches endow the Ti surface with the same antioxidant properties; (iii) that PEG-EGCg coated surfaces are more hydrophilic and show a significantly higher (>50%) interaction force with water. Obtained results build up a rationale basis for evaluation of the merits of finely tuning interfacial properties of polyphenols coated surfaces in biological tests.

Dual Rinse® HEDP increases the surface tension of NaOCl but may increase its dentin disinfection efficacy.

The aim of this study was to evaluate the surface tension and the antimicrobial activity in infected dentin of a NaOCl solution combined with an etidronate powder (Dual Rinse® HEDP), compared to pure NaOCl and the classic NaOCl + EDTA irrigating sequence, respectively. The surface tension of three irrigants was measured by Wilhelmy technique. To evaluate the antimicrobial activity of the solutions, 26 human teeth were contaminated for 5 days with E. faecalis. After bacterial contamination, ten samples were irrigated with NaOCl followed by EDTA, another ten with NaOCl/Dual Rinse® HEDP, and four were used as positive controls. Two specimens not contaminated were used as negative controls. After live/dead BacLight staining, samples were examined by CLSM for analyzing % of residual live and dead cells. Comparison of bacterial viability between and within groups was performed using the Mann-Whitney test for independent samples and the Wilcoxon signed-rank test, respectively. The mean surface tension of EDTA was significantly lower than that of the other irrigants tested (p < 0.001). Conversely, the surface tension of NaOCl/Dual Rinse® HEDP solution was significantly higher than that of all the other solutions (p < 0.001). Residual bacterial viability in the NaOCl/Dual Rinse® HEDP (1.71%) was significantly lower (p = 0.019) than in the NaOCl + EDTA group (3.77%). All of the experimental groups showed significantly lower proportion of viable bacterial cells than the positive control group (p < 0.01). Clinical relevance adding etidronate to NaOCl increases its antimicrobial effect in dentinal tubules even though increases its surface tension.

Biomimetic Surfaces Coated with Covalently Immobilized Collagen Type I: An X-Ray Photoelectron Spectroscopy, Atomic Force Microscopy, Micro-CT and Histomorphometrical Study in Rabbits.

BACKGROUND: The process of osseointegration of dental implants is characterized by healing phenomena at the level of the interface between the surface and the bone. Implant surface modification has been introduced in order to increase the level of osseointegration. The purpose of this study is to evaluate the influence of biofunctional coatings for dental implants and the bone healing response in a rabbit model. The implant surface coated with collagen type I was analyzed through X-ray Photoelectron Spectroscopy (XPS), Atomic Force Microscopy (AFM), micro-CT and histologically. METHODS: The sandblasted and double acid etched surface coated with collagen type I, and uncoated sandblasted and double acid etched surface were evaluated by X-ray Photoelectron Spectroscopy (XPS) and Atomic Force Microscopy (AFM) analysis in order evaluate the different morphology. In vivo, a total of 36 implants were positioned in rabbit articular femoral knee-joint, 18 fixtures for each surface. Micro-CT scans, histological and histomorphometrical analysis were conducted at 15, 30 and 60 days. RESULTS: A histological statistical differences were evident at 15, 30 and 60 days (p < 0.001). Both implant surfaces showed a close interaction with newly formed bone. Mature bone appeared in close contact with the surface of the fixture. The AFM outcome showed a similar roughness for both surfaces. CONCLUSION: However, the final results showed that a coating of collagen type I on the implant surface represents a promising procedure able to improve osseointegration, especially in regions with a low bone quality.

Covalently-Linked Hyaluronan versus Acid Etched Titanium Dental Implants: A Crossover RCT in Humans.

Biochemical modification of titanium surfaces (BMTiS) entails immobilization of biomolecules to implant surfaces in order to induce specific host responses. This crossover randomized clinical trial assesses clinical success and marginal bone resorption of dental implants bearing a surface molecular layer of covalently-linked hyaluronan in comparison with control implants up to 36 months after loading. Patients requiring bilateral implant rehabilitation received hyaluronan covered implants in one side of the mouth and traditional implants in the other side. Two months after the first surgery, a second surgery was undergone to uncover the screw and to place a healing abutment. After two weeks, the operator proceeded with prosthetic procedures. Implants were evaluated by periapical radiographs and the crestal bone level was recorded at mesial and distal sites-at baseline and up to 36 months. One hundred and six implants were positioned, 52 HY-coated, and 48 controls were followed up. No differences were observed in terms of insertion and stability, wound healing, implant success, and crestal bone resorption at any time considered. All interventions had an optimal healing, and no adverse events were recorded. This trial shows, for the first time, a successful use in humans of biochemical-modified implants in routine clinical practice and in healthy patients and tissues with satisfactory outcomes.

Surface chemistry and effects on bone regeneration of a novel biomimetic synthetic bone filler.

The paper presents results of physico-chemical and biological investigations of a surface-engineered synthetic bone filler. Surface analysis confirms that the ceramic phosphate granules present a collagen nanolayer to the surrounding environment. Cell cultures tests show that, in agreement with literature reports, surface-immobilized collagen molecular cues can stimulate progression along the osteogenic pathway of undifferentiated human mesenchymal cells. Finally, in vivo test in a rabbit model of critical bone defects shows statistically significant increase of bone volume and mineral apposition rate between the biomimetic bone filler and collagen-free control. All together, obtained data confirm that biomolecular surface engineering can upgrade the properties of implant device, by promoting more specific and targeted implant-host cells interactions.

Collagen type I coating stimulates bone regeneration and osteointegration of titanium implants in the osteopenic rat.

PURPOSE: To investigate the effects of titanium implants functionalised with collagen type I (TiColl) on bone regeneration and osteointegration in a healthy and osteopenic rat animal model. METHOD: TiColl screws were implanted into the femoral condyles of healthy and osteopenic rats and compared with acid-etched titanium (Ti) screws. The osteointegration process was evaluated by a complementary approach combining microtomographic, histological, histomorphometric and biomechanical investigations at four and 12 weeks. RESULTS: The TiColl screw also ensured a greater mechanical stability; the push-out values for TiColl screws increased from four to 12 weeks (+28 %). The energy necessary to detach the bone from the screw was significantly higher for TiColl-functionalised screws in comparison to Ti screws (+23 %) at 12 weeks. Histomorphometric investigation revealed that total bone-to-implant contact was higher in TiColl screws in comparison to Ti screws (P < 0.05) and at epiphyseal level, increased bone-to-implant contact was found with TiColl screws in comparison to Ti screws (P < 0.05) in an ovariectomy (OVX) condition. A significant increase in the measured total bone ingrowth from four to 12 weeks was detected for both materials, but more significant for the TiColl material (P < 0.0005). Finally, bone ingrowth in the TiColl group was significantly higher (P < 0.005) in comparison to that of Ti screws in the SHAM condition at metaphyseal level at 12 weeks. CONCLUSION: The present results showed that TiColl is effective in promoting implant osteointegration even in compromised bone.

Adherent endotoxin on dental implant surfaces: a reappraisal.

Osteoimmunology is the crosstalk between cells from the immune and skeletal systems, suggesting a role of pro-inflammatory cytokines in the stimulation of osteoclast activity. Endotoxin or bacterial challenges to inflammatory cells are directly relevant to dental implant pathologies involving bone resorption, such as osseointegration failure and peri-implantitis. While the endotoxin amount on implant devices is regulated by standards, it is unknown whether commercially available dental implants elicit different levels of adherent-endotoxin stimulated cytokines. The objective of this work is to develop a model system and evaluate endotoxin-induced expression of pro-inflammatory cytokine genes relevant to osteoclast activation on commercially available dental implants. Murine J774-A1 macrophages were cultured on Ti disks with different level of lipopolysaccharide (LPS) contamination to define the time-course of the inflammatory response to endotoxin, as evaluated by reverse transcription polymerase chain reaction analysis. The developed protocol was then used to measure adherent endotoxin on commercially available packaged and sterile dental implants in the "as-implanted" condition. Results show that tested dental implants induce variable expression of endotoxin-stimulated genes, sometimes above the level expected to promote bone resorption in vivo. Results are unaffected by the specific surface treatment; rather, they likely reflect care in cleaning and packaging protocols. In conclusion, expression of genes that enhance osteoclast activity through endotoxin stimulation of inflammatory cells is widely different on commercially available dental implants. A reappraisal of the clinical impact of adherent endotoxins on dental (and bone) implant devices is required in light of increasing knowledge on crosstalk between cells from the immune and skeletal systems.

Development of a clot-adhesive coating to improve the performance of thrombectomy devices.

BACKGROUND: The first-pass complete recanalization by mechanical thrombectomy (MT) for the treatment of stroke remains limited due to the poor integration of the clot within current devices. Aspiration can help retrieval of the main clot but fails to prevent secondary embolism in the distal arterial territory. The dense meshes of extracellular DNA, recently described in stroke-related clots, might serve as an anchoring platform for MT devices. We aimed to evaluate the potential of a DNA-reacting surface to aid the retention of both the main clot and small fragments within the thrombectomy device to improve the potential of MT procedures. METHODS: Device-suitable alloy samples were coated with 15 different compounds and put in contact with extracellular DNA or with human peripheral whole blood, to compare their binding to DNA versus blood elements in vitro. Clinical-grade MT devices were coated with two selected compounds and evaluated in functional bench tests to study clot retrieval efficacy and quantify distal emboli using an M1 occlusion model. RESULTS: Binding properties of samples coated with all compounds were increased for DNA (≈3-fold) and decreased (≈5-fold) for blood elements, as compared with the bare alloy samples in vitro. Functional testing showed that surface modification with DNA-binding compounds improved clot retrieval and significantly reduced distal emboli during experimental MT of large vessel occlusion in a three-dimensional model. CONCLUSION: Our results suggest that clot retrieval devices coated with DNA-binding compounds can considerably improve the outcome of the MT procedures in stroke patients.

Functionalization with a Polyphenol-Rich Pomace Extract Empowers a Ceramic Bone Filler with In Vitro Antioxidant, Anti-Inflammatory, and Pro-Osteogenic Properties.

Oral diseases and periodontitis in particular are a major health burden worldwide, because of their association with various systemic diseases and with conditions such as peri-implantitis. Attempts have been made over the years to reverse bone loss due to the host disproportionate inflammatory response and to prevent failure of dental implants. To this end, the use of biomaterials functionalized with molecules characterized by anti-inflammatory and antioxidant properties could represent a new frontier for regenerating functional periodontal tissues. In this study, a new ceramic granulated biomaterial, named Synergoss Red (SR), functionalized with a polyphenolic mixture extracted from pomace of the Croatina grape variety, is introduced. Following a preliminary in-depth characterization of the extract by HPLC analysis and of the biomaterial surface and composition, we performed evaluations of cytocompatibility and a biological response through in vitro assays. The anti-inflammatory and antioxidant properties of the identified phenolic molecules contained in SR were shown to downregulate inflammation in macrophages, to stimulate in osteoblast-like cells the expression of genes involved in deposition of the early bone matrix, and to mitigate bone remodeling by decreasing the RANKL/OPG ratio. Thanks to its cytocompatibility and assorted beneficial effects on bone regeneration, SR could be considered an innovative regenerative approach in periodontal therapy.

Polyphenols from grape pomace induce osteogenic differentiation in mesenchymal stem cells.

Polyphenols are increasingly investigated for the treatment of periodontitis and research on their use in dental biomaterials is currently being conducted. Grape pomace extracts are a rich source of polyphenols. In the present study, the polyphenols of two different types of grape pomace were characterized and identified by high­performance liquid chromatography­diode array detector, and the effect of polyphenol­rich grape pomace extracts on mesenchymal stem cell (MSC) osteogenic differentiation was investigated. Solid­liquid extraction was used to recover polyphenols from red and white grape pomace. The two extracts have been characterized through the phenolic content and antioxidant power. Human MSCs (hMSCs) from the bone marrow were cultured both with and without given amounts (10 or 20 µg/ml) of the obtained pomace extracts. Their effects on cell differentiation were evaluated by reverse transcription­quantitative polymerase chain reaction, compared with relevant controls. Results showed that both pomace extracts, albeit different in phenolic composition and concentration, induced multiple effects on hMSC gene expression, such as a decreased receptor activator of nuclear factor κ­Β ligand/osteoprotegerin ratio and an enhanced expression of genes involved in osteoblast differentiation, thus suggesting a shift of hMSCs towards osteoblast differentiation. The obtained results provided data in favor of the exploitation of polyphenol properties from grape pomace extracts as complementary active molecules for dental materials and devices for bone regeneration in periodontal defects.

Evaluation of Cytotoxicity and Antibacterial Activity of a New Class of Silver Citrate-Based Compounds as Endodontic Irrigants.

In the present study, the cytotoxicity and the antimicrobial activity of two silver citrate-based irrigant solutions were investigated. Cytotoxicity of various concentrations (0.25%, 0.5%, 1%, 2.5%, 5%) of both solutions (BioAKT and BioAKT Endo) was assessed on L-929 mouse fibroblasts using the MTT assay. For the quantitative analysis of components, an infrared (I.R.) spectroscopy was performed. The minimum inhibitory and minimal bactericidal concentrations (M.I.C. and M.B.C., respectively) were ascertained on Enterococcus faecalis strain ATCC 4083. For biofilm susceptibility after treatment with the irrigating agent, a minimum biofilm eradication concentration (M.B.E.C.) and confocal laser scanning microscope (C.L.S.M.) assays were performed. Quantification of E. faecalis cell biomass and percentage of live and dead cells in the biomass was appraised. Normality of data was analyzed using the D'Agostino & Pearson's test and the Shapiro-Wilk test. Statistical analysis was performed using one-way analysis of variance (ANOVA) and Tukey's test. Both silver citrate solutions showed mouse fibroblasts viability >70% when diluted to 0.25% and 0.5%. Conversely, at higher concentrations, they were extremely cytotoxic. F.T.-IR spectroscopy measurements of both liquids showed the same spectra, indicating similar chemical characteristics. No substantial contrast in antimicrobial activity was observed among the two silver citrate solutions by using broth microdilution methods, biofilm susceptibility (MBEC-HTP device), and biomass screening using confocal laser scanning microscopy (C.L.S.M.) technique. Both solutions, used as root canal irrigants, exhibited significant antimicrobial activity and low cytocompatibility at dilutions greater than 0.5%.

Development of the osteoblastic phenotype in human alveolar bone-derived cells grown on a collagen type I-coated titanium surface.

OBJECTIVE: The aim of this study was to evaluate the development of the osteoblastic phenotype in human alveolar bone-derived cells grown on collagen type I-coated titanium (Ti) surface (Col-Ti) obtained by plasma deposition acrylic acid grafting compared with machined Ti (M-Ti). MATERIAL AND METHODS: Osteoblastic cells were cultured until subconfluence and subcultured on Col-Ti and M-Ti for periods of up to 21 days. RESULTS: Cultures grown on Col-Ti and M-Ti exhibited similar cell morphology. Cell adhesion, total protein content, and alkaline phosphatase (ALP) activity were not affected by Ti surface modification in all evaluated periods. Growth analyses indicated that there were significantly more cells in cultures grown on Col-Ti at day 3. Runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteoprotegerin (OPG) mRNA expression of cells subcultured on Col-Ti was higher, whereas collagen type I (COL) was lower compared with M-Ti. Ti surface modification neither affected the osteocalcin (OC), ALP and receptor activator of NF-kappaB ligand (RANKL) mRNA expression nor the calcium content extracted from mineralized matrix. CONCLUSIONS: These results demonstrated that Col-Ti favours cell growth during the proliferative phase (day 3) and osteoblastic differentiation, as demonstrated by changes in mRNA expression profile during the matrix mineralization phase (day 14), suggesting that this Ti surface modification may affect the processes of bone healing and remodelling.

Bioactive calcium silicate ceramics and coatings.

CaO-SiO2 based ceramics have been regarded as potential candidates for artificial bone due to their excellent bone bioactivity and biocompatibility. However, they cannot be used as implants under a heavy load because of their poor mechanical properties, in particular low fracture toughness. Plasma spraying CaO-SiO2 based ceramic coatings onto titanium alloys can expand their application to the hard tissue replacement under a heavy load. Plasma sprayed wollastonite, dicalcium silicate and diopside coatings have excellent bone bioactivity and high bonding strength to titanium alloys. It is possible that these plasma sprayed CaO-SiO2 based ceramic coatings will be applied in clinic after they are widely and systematically researched.

Permanent wettability of a novel, nanoengineered, clinically available, hyaluronan-coated dental implant.

The objectives of this study are to evaluate long-term wettability of novel surface-engineered, clinically available dental implants, featuring a surface nanolayer of covalently linked hyaluronan, and to confirm the relationships between wetting properties and surface nanostructure and microstructure. Wettability measurements were performed on clinically available hyaluronan-coated Grade 4 titanium implants, packaged and sterile, that is, in the "on the shelf" condition, after 1 year from production. Wetting properties were measured by the Wilhelmy plate method. Analysis of the surface structure and chemistry was perfomed by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) analysis, atomic force microscopy (AFM), and ζ-potential measurement, either on implants or disks or plates subjected to the same surface-engineering process. Results show that hydrophilicity and ensuing capillary rise of the hyaluronan-coated implant surface is unaffected by aging and dry storage. Chemical analysis of the implant surface by XPS and evaluation of the ζ potential indicate that hyaluronan chemistry and not that of titanium dictates interfacial properties. Comparison between XPS versus EDX and SEM versus AFM data confirm that the thickness of the hyaluronan surface layer is within the nanometer range. Data show that nanoengineering of the implant surface by linking of the hydrophilic hyaluronan molecule endows tested titanium implants by permanent wettability, without need of wet storage as presently performed to keep long-term hydrophilic implant surfaces. From an analytical point of view, the introduction in routine clinical practice of nanoengineered implant surfaces requires upgrading of analytical methods to the nanoscale.

Biomolecular modification of implant surfaces.

In this review, surface modification of implant devices by immobilization of biological molecules is discussed. A brief introduction to the development of biomolecular surface science is presented, followed by a review of current activities in selected fields. Bone-contacting devices and some cardiovascular implant devices are reviewed as paradigmatic examples of research that is currently taking place. Advances in the basic fields of cell and tissue biology, in addition to concurrent developments in surface science tools, suggest that 'peri-implant biologics', or the control and direction of the host response at the implant-tissue interface by implant-surface-linked biomolecules, could be a major area of growth in the medical devices field in the next few years.

Cloning and Expression Analysis of Human Amelogenin in Nicotiana benthamiana Plants by Means of a Transient Expression System.

Enamel is the covering tissue of teeth, made of regularly arranged hydroxyapatite crystals deposited on an organic matrix composed of 90% amelogenin that is completely degraded at the end of the enamel formation process. Amelogenin has a biomineralizing activity, forming nanoparticles or nanoribbons that guide hydroxyapatite deposit, and regenerative functions in bone and vascular tissue and in wound healing. Biotechnological products containing amelogenin seem to facilitate these processes. Here, we describe the production of human amelogenin in plants by transient transformation of Nicotiana benthamiana with constructs carrying synthetic genes with optimized human or plant codons. Both genes yielded approximately 500 µg of total amelogenin per gram of fresh leaf tissue. Two purification procedures based on affinity chromatography or on intrinsic solubility properties of the protein were followed, yielding from 12 to 150 µg of amelogenin per gram of fresh leaf tissue, respectively, at different purity. The identity of the plant-made human amelogenin was confirmed by MALDI-TOF-MS analysis of peptides generated following chymotrypsin digestion. Using dynamic light scattering, we showed that plant extracts made in acetic acid containing human amelogenin have a bimodal distribution of agglomerates, with hydrodynamic diameters of 22.8 ± 3.8 and 389.5 ± 86.6 nm. To the best of our knowledge, this is the first report of expression of human amelogenin in plants, offering the possibility to use this plant-made protein for nanotechnological applications.

Low density polyethylene functionalized with antibiofilm compounds inhibits Escherichia coli cell adhesion.

The present work concerns an efficient strategy to obtain novel medical devices materials able to inhibit biofilm formation. The new materials were achieved by covalent grafting of p-aminocinnamic or p-aminosalicylic acids on low density polyethylene coupons. The polyethylene surface, previously activated by oxygen plasma treatment, was functionalized using 2-hydroxymethylmetacrylate as linker. The latter was reacted with succinic anhydride affording the carboxylic end useful for the immobilization of the antibiofilm molecules. The modified surface was characterized by scanning electron microscope, X-ray photoelectron spectroscopy, attenuated total reflectance Fourier transform infrared and fluorescence analyses. The antibiofilm activity of the modified materials were tested against Escherichia coli biofilm grown in the Center of Disease Control biofilm reactor. The results revealed that the grafted cinnamic and salicylic acid derivatives reduced biofilm biomass, in comparison with the control, by 73.7 ± 10.7% and 63.4 ± 7.1%, respectively. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3251-3261, 2017.

The effect of collagen coating on titanium with nanotopography on in vitro osteogenesis.

Several studies have shown the positive effects of Ti either with nanotopography or coated with collagen on osteoblast differentiation. Thus, we hypothesized that the association of nanotopography with collagen may increase the in vitro osteogenesis on Ti surface. Ti discs with nanotopography with or without collagen coating were characterized by scanning electron microscopy and atomic force microscopy. Rat calvaria-derived osteoblastic cells were cultured on both Ti surfaces for up to 14 days and the following parameters were evaluated: cell proliferation, alkaline phosphatase (ALP) activity, extracellular matrix mineralization, protein expression of bone sialoprotein (BSP) and osteopontin (OPN), and gene expression of collagen type 1a (Coll1a), runt-related transcription factor 2 (Runx2), osterix (OSX), osteocalcin (OC), Ki67, Survivin, and Bcl2-associated X protein (BAX). Surface characterization evidenced that collagen coating did not alter the nanotopography. Collagen coating increased cell proliferation, ALP activity, extracellular matrix mineralization, and Coll1a, OSX, OC, and BAX gene expression. Also, OPN and BSP proteins were strongly detected in cultures grown on both Ti surfaces. In conclusion, our results showed that the combination of nanotopography with collagen coating stimulates the early, intermediate, and final events of the in vitro osteogenesis and may be considered a potential approach to promote osseointegration of Ti implants. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2783-2788, 2017.

Surface tension comparison of four common root canal irrigants and two new irrigants containing antibiotic.

The aim of this study is to compare the surface tension of four common endodontic irrigants: Moltendo EDTA 17%, Cetrexidin, Smear Clear, Sodium hypochlorite 5.25%, with the surface tension of MTAD and Tetraclean. Freshly produced MilliQ water was used as a reference. All measurements were performed following the Wilhelmy plate technique, using a Cahn DCA-322 Dynamic Contact Angle Analyzer at the temperature of 22 degrees C. MilliQ water, sodium hypochlorite 5.25%, and EDTA 17% had the highest surface tension, whereas those of Cetrexedin and Tetraclean has shown the lowest surface tension value. Both new irrigants, MTAD and Tetraclean, are capable of removing the smear layer. Thanks to their low surface tension, increasing the intimate contact of irrigant solutions with the dentinal walls, they may permit deeper penetration.

In Vitro Cytokine Expression and In Vivo Healing and Inflammatory Response to a Collagen-Coated Synthetic Bone Filler.

The goal of the present work was to investigate the relationship between in vivo healing and inflammatory response and in vitro cytokine expression by macrophages of a synthetic bone filler (25% hydroxylapatite-75% ß-tricalcium phosphate) bearing a surface nanolayer of collagen. A clinically accepted, state-of-the-art xenograft material was used as a "negative control," that is, as a material that provides the correct clinical response for the intended use. In vitro data show that both materials exert a very low stimulation of proinflammatory cytokines by macrophages, and this was confirmed by the very mild inflammatory response detected in in vivo tests of local response in a rabbit model. Also, in vitro findings suggest a different mechanism of healing for the test and the control material, with a higher regenerative activity for the synthetic, resorbable filler, as confirmed by in vivo observation and literature reports. Thus, the simple in vitro model adopted provides a reasonable forecast of in vivo results, suggesting that new product development can be guided by in vitro tuning of cell-materials interactions.