Guided Tissue Regeneration Applications Research Articles

Physicochemical properties and cell proliferation and adhesive bioactivity of collagen-hyaluronate composite gradient membrane.

Membrane materials were widely used in guided tissue regeneration (GTR) to prevent fibroblast invasion and form a confined area for preferentially growing of osteoblast. A novel collagen-hyaluronate composite gradient membrane was prepared by Tilapia (Oreochromis mossambicus) skin collagen and sodium hyaluronate for potential GTR applications and their bioactivities were investigated by cellular viability. SEM results indicated the membrane showed a dense outer and a porous inner surface for effectively guiding the growth of bone tissue. Physicochemical and biosafety experiments showed the tensile strength of membrane was 466.57 ± 44.31KPa and contact angle was 74.11°, and the membrane showed perfect biocompatibility and cytocompatibility as well, which met the requirements of GTR material. Cell morphology revealed that the membrane could facilitate the adherence and proliferation of fibroblast and osteoblast. The results of qRT-PCR and ELISA demonstrated that the membrane could effectively activate TGF-β/Smad pathway in fibroblast, and promote the expressions of TGF-β1, FN1 and VEGF. Remarkably, RUNX2 was stimulated in BMP2 pathway by the membrane to regulate osteoblast differentiation. In summary, the collagen-hyaluronate composite gradient membrane not only fulfills the prerequisites for use as a GTR material but also demonstrates substantial potential for practical applications in the field.

Comparison of the Eggshell and the Porcine Pericardium Membranes for Guided Tissue Regeneration Applications.

Guided bone regeneration is frequently used to reconstruct the alveolar bone to rehabilitate the mastication using dental implants. The purpose of this article is to research the properties of eggshell membrane (ESM) and its potential application in tissue engineering. The study focuses on the structural, mechanical, and histological characteristics of ESM extracted from Gallus domesticus eggs and to compare them to a commercially available porcine pericardium membrane (Jason® membrane, botiss biomaterials GmbH, Zossen, Germany). Thus, histology was performed on the ESM, and a comparison of the microstructure through scanning electron microscopy and atomic force microscopy (AFM) was conducted. Also, mechanical tensile strength was evaluated. Samples of ESM were prepared and treated with alcohol for fixation and disinfection. Histological analysis revealed that the ESM architecture is constituted out of loose collagen fibers. However, due to the random arrangement of collagen fibers within the membrane, it might not be an effective barrier and occlusive barrier. Comparative analyses were performed between the ESM and the AFM examinations and demonstrated differences in the surface topography and mechanical properties between the two membranes. The ESM exhibited rougher surfaces and weaker mechanical cohesion attributed to its glycoprotein content. The study concludes that while the ESM displays favorable biocompatibility and resorb ability, its non-uniform collagen arrangement limits its suitability as a guided bone regeneration membrane in the current non-crosslinked native form. Crosslinking techniques may enhance its properties for such applications. Further research is needed to explore modifications and processing methods that could leverage the ESM's unique properties for tissue engineering purposes.

Fabrication and characterization of chlorhexidine gluconate loaded poly(vinyl alcohol)/45S5 nano-bioactive glass nanofibrous membrane for guided tissue regeneration applications.

Polymeric barrier membranes are used in periodontal applications to prevent fibroblastic cell migration into the cavities of bone tissue and to properly guide the proliferation of tissues. In this study, the fabrication, characterization, bioactivity, and in vitro biological properties of polyvinyl alcohol-based nanofibrous membranes containing nano-sized 45S5 bioactive glass (BG) loaded with chlorhexidine (CH) gluconate with biocompatible, bioactive, and antibacterial properties for using as dental barrier membranes were investigated. Nanofibrous membranes with an average fiber diameter, pore size, and porosity of 210 nm, 24.73 μm, and 12.42%, respectively, were loaded with 1% and 2% CH, and the release profile was investigated. The presence of BG in the membranes promoted fibroblastic proliferation and the presence of CH provided antibacterial properties. Nanofibrous membranes exhibit a high ability to restrict bacterial growth while fulfilling the necessary conditions for use as a dental barrier thanks to their low swelling rates, significant surface bioactivities, and appropriate degradation levels.

Surface Functionalization of Poly(l-lactide-co-glycolide) Membranes with RGD-Grafted Poly(2-oxazoline) for Periodontal Tissue Engineering.

Bone tissue defects resulting from periodontal disease are often treated using guided tissue regeneration (GTR). The barrier membranes utilized here should prevent soft tissue infiltration into the bony defect and simultaneously support bone regeneration. In this study, we designed a degradable poly(l-lactide-co-glycolide) (PLGA) membrane that was surface-modified with cell adhesive arginine-glycine-aspartic acid (RGD) motifs. For a novel method of membrane manufacture, the RGD motifs were coupled with the non-ionic amphiphilic polymer poly(2-oxazoline) (POx). The RGD-containing membranes were then prepared by solvent casting of PLGA, POx coupled with RGD (POx_RGD), and poly(ethylene glycol) (PEG) solution in methylene chloride (DCM), followed by DCM evaporation and PEG leaching. Successful coupling of RGD to POx was confirmed spectroscopically by Raman, Fourier transform infrared in attenuated reflection mode (FTIR-ATR), and X-ray photoelectron (XPS) spectroscopy, while successful immobilization of POx_RGD on the membrane surface was confirmed by XPS and FTIR-ATR. The resulting membranes had an asymmetric microstructure, as shown by scanning electron microscopy (SEM), where the glass-cured surface was more porous and had a higher surface area then the air-cured surface. The higher porosity should support bone tissue regeneration, while the air-cured side is more suited to preventing soft tissue infiltration. The behavior of osteoblast-like cells on PLGA membranes modified with POx_RGD was compared to cell behavior on PLGA foil, non-modified PLGA membranes, or PLGA membranes modified only with POx. For this, MG-63 cells were cultured for 4, 24, and 96 h on the membranes and analyzed by metabolic activity tests, live/dead staining, and fluorescent staining of actin fibers. The results showed bone cell adhesion, proliferation, and viability to be the highest on membranes modified with POx_RGD, making them possible candidates for GTR applications in periodontology and in bone tissue engineering.

Porous membranes of quaternized chitosan composited with strontium-based nanobioceramic for periodontal tissue regeneration.

This report demonstrates the development of a degradable quaternary ammonium derivative of chitosan (QC) composited with strontium-containing nanoapatite (SA) for bioactivity. The material was made as porous membrane by solution casting and freeze drying, for guided tissue regeneration (GTR) applications. The micromorphology, tensile strength, suture pull-out strength, degradation (in vitro, in phosphate buffered saline), and cytocompatibility (using human periodontal ligament cells) were tested to investigate the effect of derivatization and SA addition. The porosity of the membranes increased with increasing SA content and so did the tensile strength and the degradation. The suture pull-out strength, however, showed a decrease. The cell culture evaluation endorsed biocompatibility. The composite with 1.5mg SA per 1mL QC was found to have optimal qualities for GTR applications.

Controlled release of bFGF loaded into electrospun core–shell fibrous membranes for use in guided tissue regeneration

In this study, basic fibroblast growth factor (bFGF) was loaded into a poly(lactic-co-glycolic acid) (PLGA)/wool keratin composite membrane by emulsion electrospinning to prepare a composite membrane with a core–shell structure for guided tissue regeneration (GTR). The physicochemical properties, drug release performance in vitro and cytotoxicity of the composite membrane were evaluated to select the optimum concentration of bFGF. The fibers in the six groups of composite membranes were uniform in thickness, had a smooth surface, and did not exhibit a string bead structure. In addition, the fibers in the six groups of composite membranes had a stable core–shell structure, in which the core layer of the fibers was wrapped around bFGF and the shell layer of the fibers comprised the PLGA/wool keratin oil-phase component. Compared with the PLGA/wool keratin composite membrane prepared by traditional electrospinning, the composite membranes prepared by emulsion electrospinning had a higher water absorption rate and superior hydrophilicity. The bFGF encapsulation rate of the 10 μg bFGF composite membrane was the highest (97.3% ± 11.3%). Stable and sustained release of bFGF from the five groups of bFGF-loaded PLGA/wool keratin composite membranes can be maintained over 28 d. The five groups of PLGA/wool keratin composite membranes loaded with bFGF could promote the adhesion, proliferation and osteogenic differentiation of human periodontal ligament fibroblasts (hPLDFs) to varying degrees. Among the five groups, the cell morphology, proliferation ability and osteogenic differentiation ability of the 10 μg bFGF composite membrane group were the best. This study will serve as a foundation for further research on bFGF-loaded composite membranes for GTR applications.

Development and characterisation of bilayered periosteum-inspired composite membranes based on sodium alginate-hydroxyapatite nanoparticles

Background and aimMembranes for guided bone regeneration should have a mechanical structure and a chemical composition suitable for mimicking biological structures. In this work, we pursue the development of periosteum-inspired bilayered membranes obtained by crosslinking alginate with different amounts of nanohydroxyapatite. ExperimentsAlginate-nanohydroxyapatite interaction was studied by rheology and infrared spectroscopy measurements. The membranes were characterized regarding their tensile strength, degradation and surface morphology. Finally, cell cultures were performed on each side of the membranes. FindingsThe ionic bonding between alginate polysaccharide networks and nanohydroxyapatite was proven, and had a clear effect in the strength and microstructure of the hydrogels. Distinct surface characteristics were achieved on each side of the membranes, resulting in a highly porous fibrous side and a mineral-rich side with higher roughness and lower porosity. Moreover, the effect of amount of nanohydroxyapatite was reflected in a decrease of the membranes’ plasticity and an increment of degradation rate. Finally, it was proved that osteoblast-like cells proliferated and differentiated on the mineral-rich side, specially when a higher amount of nanohydroxyapatite was used, whereas fibroblasts-like cells were able to proliferate on the fibrous side. These periosteum-inspired membranes are promising biomaterials for guided tissue regeneration applications.

Bioinspired and osteopromotive polydopamine nanoparticle-incorporated fibrous membranes for robust bone regeneration

Due to their inherent hydrophobic and bioinert nature, synthetic degradable polymer-based membranes show inferior stem cell attachment, proliferation, and even differentiation. To overcome these limitations, bioinspired and osteopromotive polydopamine nanoparticle-incorporated fibrous membranes are developed via a two-step route: pH-induced polymerization of dopamine and co-electrospinning of polycaprolactone (PCL) with polydopamine nanoparticles (PDA NPs). Hybrid membranes with optimized PDA NP content exhibit high quantities of apatite deposition and prominent cytocompatibility (cell attachment, spreading and reproduction) and osteo-differentiation potential (alkaline phosphatase activity, calcium mineralization, and osteogenesis-related genes and protein expression) of human mesenchymal stem cells cultured without any growth factors. Importantly, in vivo assessments using a mouse calvarial critical-sized defect demonstrate that the engineered fibrous membranes remarkably boost bone reconstruction and regeneration. Accordingly, our bioinspired PCL-based hybrid fibrous membranes with robust osteoinductive ability can potentially be utilized as a clinically applicable candidate in guided tissue regeneration applications.

Extracellular Matrix Membrane Induces Cementoblastic/Osteogenic Properties of Human Periodontal Ligament Stem Cells.

Objective: Periodontitis affects nearly 90% of adults over the age of 70, resulting to periodontal tissue infection, destruction, and ultimately tooth loss. Guided tissue regeneration (GTR) is a method widely used to treat severe periodontal disease, and involves placement of an occlusive barrier to facilitate regeneration of the damaged area by periodontal ligament stem cells (PDLSCs). In this study, we evaluate natural extracellular matrix (ECM) as a scaffold material to provide a suitable microenvironment to support the proliferation, differentiation, and tissue-regenerating properties of PDLSCs.Design: The viability, proliferation, apoptosis, and migration of PDLSCs cultured on ECM membrane, that was isolated from porcine urinary bladders, were compared with those cultured on type I collagen membrane, a commonly used scaffold in GTR. To evaluate the effects of ECM vs. type I collagen on the tissue-regenerating properties of PDLSCs, the bio-attachment and cementoblastic/osteogenic differentiation of PDLSCs were evaluated.Results: Incubation of PDLSCs with ECM resulted in increased viability, proliferation, and reduced apoptosis, compared with type I collagen treated PDLSCs. Co-culture with ECM membrane also increased the migration and bio-attachment of PDLSCs. Incubation of PDLSCs with ECM membrane increased expression of the cementoblastic/osteogenic differentiation markers BSP, RUNX2, ALP, OPN, OCN, and periostin.Conclusion: ECM membrane enhances the proliferation and regenerative properties of PDLSCs, indicating that ECM membrane can serve as a suitable scaffold in the application of GTR to treat periodontal disease.

Biodegradable, Biomimetic Elastomeric, Photoluminescent, and Broad-Spectrum Antibacterial Polycitrate-Polypeptide-based Membrane toward Multifunctional Biomedical Implants.

Current biomedical membranes in guided tissue regeneration applications are almost nonbiodegradable or deficient in functionality. The development of biodegradable biomaterials with multifunctional properties including biomimetic elastomeric behavior, self-anti-infection, noninvasive monitoring, and good biocompatibility has attracted much attention. Here, we report a biodegradable and biocompatible polycitrate-(ε-polypeptide)-based (PCE) biomedical elastomer membrane with intrinsic broad-spectrum antibacterial activity and photoluminescent capacity for multifunctional guided tissue regenerative applications. PCE showed highly elastomeric mechanical behavior (∼300% elongation and ∼100% recovery) and biomimetic mechanical properties against several native tissues. PCE film also possessed highly efficient broad-spectrum antibacterial activity in vitro and in vivo. The inherent photoluminescent properties of PCE film endowed their real-time noninvasive monitoring capacity in vivo. Owing to the biocompatible structure (polycitrate and natural polypeptide), PCE film demonstrated significantly high cytocompatibility and hemocompatibility in vitro and low inflammatory response in vivo. Our study may provide a new strategy to design next generation multifunctional biodegradable biomedical implant membranes for smart guided tissue regenerative medicine applications.

3D-printed membrane for guided tissue regeneration

Three-dimensional (3D) printing is currently being intensely studied for a diverse set of applications, including the development of bioengineered tissues, as well as the production of functional biomedical materials and devices for dental and orthopedic applications. The aim of this study was to develop and characterize a 3D-printed hybrid construct that can be potentially suitable for guided tissue regeneration (GTR). For this purpose, the rheology analyses have been performed on different bioinks and a specific solution comprising 8% gelatin, 2% elastin and 0.5% sodium hyaluronate has been selected as the most suitable composition for printing a structured membrane for GTR application. Each membrane is composed of 6 layers with strand angles from the first layer to the last layer of 45, 135, 0, 90, 0 and 90°. Confirmed by 3D Laser Measuring imaging, the membrane has small pores on one side and large pores on the other to be able to accommodate different cells like osteoblasts, fibroblasts and keratinocytes on different sides. The ultimate cross-linked product is a 150μm thick flexible and bendable membrane with easy surgical handling. Static and dynamic mechanical testing revealed static tensile modules of 1.95±0.55MPa and a dynamic tensile storage modulus of 314±50kPa. Through seeding the membranes with fibroblast and keratinocyte cells, the results of in vitro tests, including histological analysis, tissue viability examinations and DAPI staining, indicated that the membrane has desirable in vitro biocompatibility. The membrane has demonstrated the barrier function of a GTR membrane by thorough separation of the oral epithelial layer from the underlying tissues. In conclusion, we have characterized a biocompatible and bio-resorbable 3D-printed structured gelatin/elastin/sodium hyaluronate membrane with optimal biostability, mechanical strength and surgical handling characteristics in terms of suturability for potential application in GTR procedures.

Evaluation of Epigallocatechin-3-gallate Modified Collagen Membrane and Concerns on Schwann Cells.

Collagen is an essential component of the extracellular matrix (ECM) and is a suitable material for nerve repair during tissue remodeling for fracture repair. Epigallocatechin-3-gallate (EGCG), an extract of green tea, shows various biological activities that are beneficial to nerve repair. Here, we developed modified collagen containing different concentrations of EGCG (0.0064%, 0.064%, and 0.64%, resp.) to induce Schwann cell proliferation and differentiation. Cell Counting Kit-8 test, live/dead assay, and SEM showed that collagen cross-linked by EGCG induced Schwann cell proliferation. Real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting revealed that EGCG-modified collagen induced Schwann cell differentiation and downregulated reactive oxygen species (ROS) levels by downregulating the MAPK P38 signaling pathway. Our results indicate that collagen cross-linked with an appropriate concentration of EGCG induces the proliferation and differentiation of Schwann cells. The EGCG-modified collagen membrane may be applicable for nerve repair and guided tissue regeneration applications.

Protein adsorption capability on polyurethane and modified-polyurethane membrane for periodontal guided tissue regeneration applications

Periodontal disease if left untreated can result in creation of defects within the alveolar ridge. Barrier membranes are frequently used with or without bone replacement graft materials for achieving periodontal guided tissue regeneration (GTR). Surface properties of barrier membranes play a vital role in their functionality and clinical success. In this study polyetherurethane (PEU) membranes were synthesized by using 4,4′-methylene-diphenyl diisocyanate (MDI), polytetramethylene oxide (PTMO) and 1,4-butane diol (BDO) as a chain extender via solution polymerization. Hydroxyl terminated polydimethylsiloxane (PDMS) due to having inherent surface orientation towards air was used for surface modification of PEU on one side of the membranes. This resulting membranes had one surface being PEU and the other being PDMS coated PEU. The prepared membranes were treated with solutions of bovine serum albumin (BSA) in de-ionized water at 37°C at a pH of 7.2. The surface protein adsorptive potential of PEU membranes was observed using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Raman spectroscopy and Confocal Raman spectroscopy. The contact angle measurement, tensile strength and modulus of prepared membranes were also evaluated. PEU membrane (89.86±1.62°) exhibited less hydrophobic behavior than PEU-PDMS (105.87±3.16°). The ultimate tensile strength and elastic modulus of PEU (27±1MPa and 14±2MPa) and PEU-PDMS (8±1MPa and 26±1MPa) membranes was in required range. The spectral analysis revealed adsorption of BSA proteins on the surface of non PDMS coated PEU surface. The PDMS modified PEU membranes demonstrated a lack of BSA adsorption. The non PDMS coated side of the membrane which adsorbs proteins could potentially be used facing towards the defect attracting growth factors for periodontal tissue regeneration. Whereas, the PDMS coated side could serve as an occlusive barrier for preventing gingival epithelial cells from proliferating and migrating into the defect space by facing the soft tissue flaps. This study demonstrates the potential of a dual natured PEU barrier membrane for use in periodontal tissue engineering applications and further investigations are required.

Bioactivity, biocompatibility and antimicrobial properties of a chitosan-mineral composite for periodontal tissue regeneration

A composite membrane of the polymer, chitosan, and the silver-exchanged mineral phase, tobermorite, was prepared by solvent casting and characterised by scanning electron microscopy and Fourier transform infrared spectroscopy. The in vitro bioactivity, cytocompatibility and antimicrobial activity of the composite were evaluated with respect to its potential application as a guided tissue regeneration (GTR) membrane. The in vitro bioactivity was verified by the formation of hydroxyapatite on the surface of the membrane in simulated body fluid and its cytocompatibility was established using MG63 human osteosarcoma cells. The presence of silver ions conferred significant antimicrobial activity against S. aureus, P. aeruginosa and E. coli. The findings of this investigation have indicated that the chitosan-silver-tobermorite composite is a prospective candidate for GTR applications.

Freeze gelated porous membranes for periodontal tissue regeneration

Guided tissue regeneration (GTR) membranes have been used for the management of destructive forms of periodontal disease as a means of aiding regeneration of lost supporting tissues, including the alveolar bone, cementum, gingiva and periodontal ligaments (PDL). Currently available GTR membranes are either non-biodegradable, requiring a second surgery for removal, or biodegradable. The mechanical and biofunctional limitations of currently available membranes result in a limited and unpredictable treatment outcome in terms of periodontal tissue regeneration. In this study, porous membranes of chitosan (CH) were fabricated with or without hydroxyapatite (HA) using the simple technique of freeze gelation (FG) via two different solvents systems, acetic acid (ACa) or ascorbic acid (ASa). The aim was to prepare porous membranes to be used for GTR to improve periodontal regeneration. FG membranes were characterized for ultra-structural morphology, physiochemical properties, water uptake, degradation, mechanical properties, and biocompatibility with mature and progenitor osteogenic cells. Fourier transform infrared (FTIR) spectroscopy confirmed the presence of hydroxyapatite and its interaction with chitosan. μCT analysis showed membranes had 85–77% porosity. Mechanical properties and degradation rate were affected by solvent type and the presence of hydroxyapatite. Culture of human osteosarcoma cells (MG63) and human embryonic stem cell-derived mesenchymal progenitors (hES-MPs) showed that all membranes supported cell proliferation and long term matrix deposition was supported by HA incorporated membranes. These CH and HA composite membranes show their potential use for GTR applications in periodontal lesions and in addition FG membranes could be further tuned to achieve characteristics desirable of a GTR membrane for periodontal regeneration.

Preparation and Bioactivity Properties of a Novel Composite Membrane of Fructose Mediatedβ-Tricalcium Pyrophosphate/(Polyethylene Glycol/Chitosan) for Guided Tissue Regeneration

A novel composite membrane ofβ-tricalcium pyrophosphate (β-TCP) and fructose- (F-) mediated chitosan/poly(ethylene glycol) (CS/PEG) was prepared by thermally induced phase separation technique. The prepared composite membranes were characterized using scanning electron microscopy (SEM) and X-ray diffraction (XRD). The mechanical property, swelling, degradation, and cytotoxicity of the composite membranes were evaluated in vitro with respect to its potential for use as biodegradable guided tissue regeneration (GTR) membrane. In vitro degradation tests showed the composite membrane with a controllable degradation rate when changing theβ-TCP content. The incorporation ofβ-TCP granules also caused a significant enhancement of tensile strength. Whenβ-TCP content is controlled to 50 wt%, homogeneous composite membranes with well mechanical property and enzymatic degradation rate can be obtained. Cytotoxicity assay demonstrates that the composite membranes were nontoxic and had very good cell compatibility. Most importantly, the release of calcium ions and glucosamine from the composite membranes was proved to increase the cell proliferation of NIH3T3. The results of this study have indicated that this novel F-β-TCP/CS/PEG composite can be a suitable material for GTR applications.

PCL-forsterite nanocomposite fibrous membranes for controlled release of dexamethasone.

The well-known treatment of the alveolar bone defects is guided tissue regeneration (GTR). Engineered membranes combined with osteo-differentiation factors have been offered a promising strategy for GTR application. Recently, poly(ε-caprolactone) (PCL)-forsterite (PCL-F) nanocomposite fibrous membranes have been developed. However, PCL-F membranes could not promote bone tissue regeneration. The aim of this research is to encapsulate an osteogenic factor [dexamethasone (DEX)] in PCL-F membranes and evaluate the effects of forsterite nanopowder (particle size = 25-45 nm) and fiber organization on DEX delivery for GTR application. The hypothesis is that the release kinetic and profile of DEX could be controlled through variation of forsterite content (0, 5 and 10 wt%) and fiber arrangement (aligned and random). Results demonstrated while DEX release was sustained over a period of 4 weeks, its kinetic was governed by the membrane architecture and composition. For example, aligned PCL-F nanocomposite fibrous membrane consisting of 10 %(w/v) forsterite nanopowder exhibited the least initial burst release (13 % release in the first 12 h) and allowed sustained release of DEX. Additionally, forsterite nanopowder inclusion changed the kinetic of DEX release from Fickian diffusion to an anomalous transport. The bioactivity of released DEX was estimated using culturing the stem cells from human exfoliated deciduous teeth (SHED) on the membranes. Results demonstrated that proliferation and osteogenic differentiation of SHED could be governed by DEX release process. While DEX release from the membranes decreased SHED proliferation, stimulated the matrix mineralization. Our finding indicated that aligned PCL-F/DEX membrane could be used as a carrier for the sustained release of drugs relevant for GTR trophy.

Antibacterial Activity of a Chitosan-PVA-Ag+-Tobermorite Composite for Periodontal Repair

A polymer-mineral composite was prepared by solvent casting a mixture of chitosan, poly(vinyl alcohol), and Ag+-exchanged tobermorite in dilute acetic acid and characterised by scanning electron microscopy and Fourier transform infrared spectroscopy. The in vitro bioactivity of the CPTAg membrane was confirmed by the formation of hydroxyapatite on its surface in simulated body fluid. The alkaline dissolution products of the tobermorite lattice buffered the acidic breakdown products of the chitosan polymer and the presence of silver ions resulted in marked antimicrobial action against S. aureus, P. aeruginosa, and E. coli. The in vitro cytocompatibility of the CPTAg membrane was confirmed using MG63 osteosarcoma cells. The findings of this preliminary study have indicated that chitosan-poly(vinyl alcohol)-Ag+-tobermorite composites may be suitable materials for guided tissue regeneration applications.

Bioactivity and biocompatibility of a chitosan-tobermorite composite membrane for guided tissue regeneration

A polymer-mineral composite membrane of the mucopolysaccharide derivative, chitosan, and calcium silicate hydrate phase, tobermorite, was prepared by solvent casting and characterised by scanning electron microscopy (SEM) and Fourier Transform infrared spectroscopy (FTIR). The bioactivity and biocompatibility of the chitosan-tobermorite composite were evaluated in vitro with respect to its potential for use as a biodegradable guided tissue regeneration (GTR) membrane. The in vitro bioactivity of the composite was confirmed by the formation of crystalline substituted hydroxyapatite on the surface of the embedded tobermorite particles in simulated body fluid. The presence of the composite membrane was found to enhance the growth of MG63 human osteosarcoma cells by up to 30%. The findings of this initial study have indicated that this novel chitosan-tobermorite composite may be a suitable material for GTR applications.

Suture pullout strength and in vitro fibroblast and RAW 264.7 monocyte biocompatibility of genipin crosslinked nanofibrous chitosan mats for guided tissue regeneration

Guided tissue regeneration (GTR) is a surgical technique used to direct the formation of bone in the graft space by protecting it with a barrier membrane used to exclude soft tissues during healing. Chitosan has been advocated for GTR applications because of its biocompatibility, degradability, wound healing, and osteogenic properties. In this study, electrospun chitosan membranes, crosslinked with 5 mM or 10 mM geinipin, a natural crosslinker extracted from the gardenia plant, were evaluated for suture pullout strength, crystallinity, and cytocompatibility with normal human dermal fibroblast and TIB 71™ RAW 264.7 monocyte cells. Ultimate suture pullout strength was significantly lower (51-67%) than that of commercially available collagen membranes. Crystallinity of the electrospun chitosan mats decreased upon crosslinking by 14-17% (p = 0.013). The molecular weight of the chitosan polymer was decreased by 75% during the electrospinning process. Uncrosslinked and genipin-crosslinked chitosan mats were cytocompatible and supported fibroblast cell proliferation for 9 days. Uncrosslinked and genipin-crosslinked membranes did not activate monocytes to produce nitric oxide (NO) in vitro in the absence of lipopolysaccharide (LPS). Finally, chitosan membranes inhibited LPS-induced NO production of RAW 264.7 cells by 59-67% as compared to tissue culture plastic and collagen membrane. Improvements are needed in the tear strength of electrospun chitosan membranes for clinical application.