Evaluation of in vitro anti-inflammatory effects of crude ginger and rosemary extracts obtained through supercritical CO2 extraction on macrophage and tumor cell line: the influence of vehicle type.

BACKGROUND: Numerous plants from have been investigated due to their anti-inflammatory activity and, among then, extracts or components of ginger (Zingiber officinale Roscoe) and rosemary (Rosmarinus officinalis L.), sources of polyphenolic compounds. 6-gingerol from ginger rhizome and carnosic acid and carnosol from rosemary leaves present anti-tumor, anti-inflammatory and antioxidant activities. However, the evaluation of the mechanisms of action of these and other plant extracts is limited due to their high hydrophobicity. Dimethylsulfoxide (DMSO) is commonly used as a vehicle of liposoluble materials to mammalian cells in vitro, presenting enhanced cell penetration. Liposomes are also able to efficiently deliver agents to mammalian cells, being capable to incorporate in their structure not only hydrophobic molecules, but also hydrophilic and amphiphilic compounds. Another strategy is based on the use of Pluronic F-68, a biocompatible low-foaming, non-ionic surfactant, to disperse hydrophobic components. Here, these three delivery approaches were compared to analyze their influence on the in vitro anti-inflammatory effects of ginger and rosemary extracts, at different concentrations, on primary mammalian cells and on a tumor cell line. METHODS: Ginger and rosemary extracts free of organic solvents were obtained by supercritical fluid extraction and dispersed in DMSO, Pluronic F-68 or liposomes, in variable concentrations. Cell viability, production of inflammatory mediators and nitric oxide (NO) release were measured in vitro on J774 cell line and murine macrophages primary culture stimulated with bacterial lipopolysaccharide and interferon-γ after being exposed or not to these extracts. RESULTS: Ginger and rosemary extracts obtained by supercritical CO2 extraction inhibited the production of pro-inflammatory cytokines and the release of NO by peritoneal macrophages and J774 cells. The delivery vehicles influenced the anti-inflammatory effects. Comparatively, the ginger extract showed the highest anti-inflammatory activity on the tumor cell line. Controversially, rosemary extract dispersed on DMSO induced a more significant IL-1 and TNF-α reduction than ginger extract in primary macrophages. CONCLUSIONS: Amongst the tested delivery vehicles, DMSO was the most suitable, presenting reduced cytotoxicity, followed by Pluronic F-68 and liposomes, provably due to differences in their form of absorption, distribution and cellular metabolism. Co-administration of liposomes and plant extracts may cause death of macrophages cells and induction of NO production. It can be concluded that some of the beneficial effects attributed to extracts of ginger and rosemary may be associated with the inhibition of inflammatory mediators due to their high antioxidant activity. However, these effects were influenced by the type of delivery vehicle.

Analysis of Key Factors for Evaluating Mucosal Adhesion Using Swine Buccal Tissue.

The assessment of the mucoadhesive properties peak mucoadhesive force (Fmax) and work of mucoadhesion (Wmuc) with texture analyzers is a common in vitro method for analyzing formulation capabilities. Challenges arise in selecting and standardizing experimental conditions due to various variables influencing mucoadhesion. This complexity hampers direct product performance comparisons. In our study, we explored factors (contact force and time, probe speed and mucin in artificial saliva) impacting a model formulation's mucoadhesive capacity. Using Omcilon-A®Orabase on porcine buccal mucosa, we systematically varied experimental conditions, employing a statistical approach (Central Composite Design - CCD). Three variables (contact force, contact time, probe speed) and their interactions were assessed for their impact on Fmax and Wmuc. Results showed that contact time and force positively affected Fmax, while only contact time influenced Wmuc. In the mucin artificial saliva test, a force of 0.5 N, time of 600 s, and speed of 1 mm/s yielded optimal Fmax (0.587 N) and Wmuc (0.468 N.s). These conditions serve as a reference for comparing mucoadhesive properties of formulations for topical oral use.

Cell-Friendly Chitosan-Xanthan Gum Membranes Incorporating Hydroxyapatite Designed for Periodontal Tissue Regeneration.

In this work, a simple method was proposed to produce dense composite polysaccharide-based membranes to be used for guided tissue and guided bone regeneration. The mucoadhesive polysaccharides chitosan (C) and xanthan gum (X) were used to produce polyelectrolyte-based complex membranes. Hydroxyapatite (HA) was added to the formulation as a potential drug carrier, in C:X:HA mass proportions equal to 1:1:0.4, 1:1:2, and 1:1:10, and also to improve membranes bioactivity and biomimetic properties. FTIR analysis indicated successful incorporation of HA in the membranes and XRD analysis showed that no changes in the HA crystalline structure were observed after incorporation. The residual mass evaluated by TGA was higher for the formulation produced at the proportion 1:1:10. The membranes produced showed asymmetrical surfaces, with distinct roughness. Increasing the HA concentration increased the surface roughness. Greater in vitro proliferation of dental pulp mesenchymal stem cells was observed on the surface of the membrane with 1:1:10 C:X:HA proportion. However, the 1:1:2 formulation showed the most adequate balance of mechanical and biological properties. These results suggest that adding HA to the membranes can influence mechanical parameters as well as cell adhesion and proliferation, supporting the potential application of these materials in regenerative techniques and the treatment of periodontal lesions.

Human dental pulp stem cell monolayer and spheroid therapy after spinal motor root avulsion in adult rats.

Spinal cord injuries result in severe neurological deficits and neuronal loss, with poor functional recovery. Mesenchymal stem cells have shown promising results; therefore the present objective of this work was to compare motor recovery after treatment with human dental pulp stem cells (hDPSC) cultivated in monolayer (2D) or as spheroids (3D), following avulsion and reimplantation of spinal motor roots in adult rats. Thus, 72 adult female Lewis rats were divided into 4 groups: avulsion (AV); avulsion followed by reimplantation (AR); avulsion associated with reimplant and 2D cell therapy (AR + 2D), and avulsion associated with reimplant and 3D cell therapy (AR + 3D). The application of the cells in 2D and 3D was performed by microsurgery, with subsequent functional assessment using a walking track test (Catwalk system), immunohistochemistry, neuronal survival, and qRT-PCR in 1-, 4-, and 12-weeks post-injury. The animals in the AR + 2D and AR + 3D groups showed the highest neuronal survival rates, and immunofluorescence revealed downregulation of GFAP, and Iba-1, with preservation of synaptophysin, indicating a reduction in glial reactivity, combined with the maintenance of pre-synaptic inputs. There was an increase in anti-inflammatory (IL-4, TGFß) and a reduction of pro-inflammatory factors (IL-6, TNFα) in animals treated with reimplantation and hDPSC. As for the functional recovery, in all analyzed parameters, the AR + 2D group performed better and was superior to the avulsion alone. Overall, our results indicate that the 2D and 3D cell therapy approaches provide successful immunomodulation and motor recovery, consistent with advanced therapies after spinal cord injury.

Polymeric Biomaterials for Topical Drug Delivery in the Oral Cavity: Advances on Devices and Manufacturing Technologies.

There are several routes of drug administration, and each one has advantages and limitations. In the case of the topical application in the oral cavity, comprising the buccal, sublingual, palatal, and gingival regions, the advantage is that it is painless, non-invasive, allows easy application of the formulation, and it is capable of avoiding the need of drug swallowing by the patient, a matter of relevance for children and the elderly. Another advantage is the high permeability of the oral mucosa, which may deliver very high amounts of medication rapidly to the bloodstream without significant damage to the stomach. This route also allows the local treatment of lesions that affect the oral cavity, as an alternative to systemic approaches involving injection-based methods and oral medications that require drug swallowing. Thus, this drug delivery route has been arousing great interest in the pharmaceutical industry. This review aims to condense information on the types of biomaterials and polymers used for this functionality, as well as on production methods and market perspectives of this topical drug delivery route.

Development of a device useful to reproducibly produce large quantities of viable and uniform stem cell spheroids with controlled diameters.

Three-dimensional cellular aggregates can mimic the natural microenvironment of tissues and organs and obtaining them through controlled and reproducible processes is mandatory for scaling up and implementing drug cytotoxicity and efficacy tests, as well as tissue engineering protocols. The purpose of this work was to develop and evaluate the performance of a device with two different geometries fabricated by additive manufacturing. The methodology was based on casting a microwell array insert using a non-adhesive hydrogel to obtain highly regular microcavities to standardize spheroid formation and morphology. Spheroids of dental pulp stem cells, bone marrow stromal cells and embryonic stem cells showing high cell viability and average diameters of around 253, 220, and 500 µm, respectively, were produced using the device with the geometry considered most adequate. The cell aggregates showed sphericity indexes above 0.9 and regular surfaces (solidity index higher than 0.96). Around 1000 spheroids could be produced in a standard six-well plate. Overall, these results show that this method facilitates obtaining a large number of uniform, viable spheroids with pre-specified average diameters and through a low-cost and reproducible process for a myriad of applications.

Comparative study on complexes formed by chitosan and different polyanions: Potential of chitosan-pectin biomaterials as scaffolds in tissue engineering.

Polyelectrolyte complexes of chitosan (Ch) and pectin (Pc) or alginate (Alg) were produced in the presence or absence of the silicone gel Silpuran® 2130 A/B (Sil) and the surfactant Kolliphor® P188 (Kol). Ch-Pc-Kol-based formulations presented higher porosity (up to 83.3%) and thickness (maximum of 2273.5 µm in PBS). Lower water contact angle was observed for Ch-Alg formulations (minimum of 36.8°) and these formulations presented higher swelling and mass loss in PBS (reaching up to 21.7 g/g and 80.4%, respectively). The addition of Sil to the matrices improved their elastic moduli, reaching a maximum of 4-fold change at 40% strain. The use of pectin instead of alginate augmented the elastic moduli, reaching 66 and 4-fold changes for dense and porous formulations, respectively. Pectin-containing scaffolds presented poroviscoelasticity, a typical mechanical feature of many soft tissues. The suitability of the materials for tissue engineering applications was demonstrated in terms of stability upon degradation in culture medium or lysozyme solution, as well as lack of cytotoxicity. This study evidences the potential of Ch-Pc-based materials to be further explored for this purpose, especially to improve the mechanical properties of chitosan-based scaffolds aiming medical applications.

Coated electrospun bioactive wound dressings: Mechanical properties and ability to control lesion microenvironment.

Advanced wound dressings capable of interacting with lesions and changing the wound microenvironment to improve healing are promising to increase the therapeutic efficacy of this class of biomaterials. Aiming at the production of bioactive wound dressings with the ability to control the wound microenvironment, biomaterials of three different chemical compositions, but with the same architecture, were produced and compared. Electrospinning was employed to build up a biomimetic extracellular matrix (ECM) layer consisting of poly(caprolactone) (PCL), 50/50 dl-lactide/glycolide copolymer (PDLG) and poly(l-lactide) (PLLA). As a post-treatment to broaden the bioactivity of the dressings, an alginate coating was applied to sheathe and functionalize the surface of the hydrophobic electrospun wound dressings, in combination with the extract of the plant Arrabidaea chica Verlot, known for its anti-inflammatory and healing promotion properties. Wettable bioactive structures capable to interact with media simulating lesion microenvironments, with tensile strength and elongation at break ranging respectively from 155 to 273 MPa and from 0.94 to 1.39% were obtained. In simulated exudative microenvironment, water vapor transmission rate (WVTR) values around 700 g/m2/day were observed, while water vapor permeability rates (WVPR) reached about 300 g/m2/day. In simulated dehydrated microenvironment, values of WVTR around 200 g/m2/day and WVPR around 175 g/m2/day were attained.

Polysaccharide-based tissue-engineered vascular patches.

Coronary artery and peripheral vascular diseases are the leading cause of morbidity and mortality worldwide and often require surgical intervention to replace damaged blood vessels, including the use of vascular patches in endarterectomy procedures. Tissue engineering approaches can be used to obtain biocompatible and biodegradable materials directed to this application. In this work, dense or porous scaffolds constituted of chitosan (Ch) complexed with alginate (A) or pectin (P) were fabricated and characterized considering their application as tissue-engineered vascular patches. Scaffolds fabricated with alginate presented higher culture medium uptake capacity (up to 17 g/g) than materials produced with pectin. A degradation study of the patches in the presence of lysozyme showed longer-term stability for Ch-P-based scaffolds. Pectin-containing matrices presented higher elastic modulus (around 280 kPa) and ability to withstand larger deformations. Moreover, these materials demonstrated better performance when tested for hemocompatibility, with lower levels of platelet adhesion and activation. Human smooth muscle cells (HSMC) adhered, spread and proliferated better on matrices produced with pectin, probably as a consequence of cell response to higher stiffness of this material. Thus, the outcomes of this study demonstrate that Ch-P-based scaffolds present superior characteristics for the application as vascular patches. Despite polysaccharides are yet underrated in this field, this work shows that biocompatible tridimensional structures based on these polymers present high potential to be applied for the reconstruction and regeneration of vascular tissues.

Influence of the incorporation of the antimicrobial agent polyhexamethylene biguanide on the properties of dense and porous chitosan-alginate membranes.

This work is a continuation of a previous study which described the development of dense and porous chitosan-alginate polyelectrolyte complexes through the addition of different amounts of Pluronic F68 to the polymeric mixture. The present study consisted in the incorporation of an antimicrobial agent, polyhexamethylene biguanide (PHMB), to the previously developed system. PHMB was incorporated at 1 and 10% (w/w) with high incorporation efficiencies, varying from 72 to 86%. Release profiles in phosphate buffered saline were evaluated using the Korsmeyer-Peppas equation, which suggested a quasi-Fickian diffusion mechanism for all obtained formulations. The maximum release percentage was approximately 15% as a result from the high affinity between PHMB and the polysaccharides. The obtained polyelectrolyte complexes were able to prevent the growth of both Staphylococcus aureus and Pseudomonas aeruginosa on their surfaces, being considered potentially effective wound dressings.

Effects of chitosan solution concentration and incorporation of chitin and glycerol on dense chitosan membrane properties.

The aim of this work was to perform a systematic study about the effects induced by chitosan solution concentration and by chitin or glycerol incorporation on dense chitosan membranes with potential use as burn dressings. The membrane properties analyzed were total raw material cost, thickness, morphology, swelling ratio, tensile strength, percentage of strain at break, crystallinity, in vitro enzymatic degradation with lysozyme, and in vitro Vero cells adhesion. While the use of the most concentrated chitosan solution (2.5% w/w) increased membrane cost, it also improved the biomaterial mechanical resistance and ductility, as well as reduced membrane degradation when exposed for 2 months to lysozyme. The remaining evaluated properties were not affected by initial chitosan solution concentration. Chitin incorporation, on the other hand, reduced the membranes cost, swelling ratio, mechanical properties, and crystallinity, resulting in thicker biomaterials with irregular surface more easily degradable when exposed to lysozyme. Glycerol incorporation also reduced the membranes cost and crystallinity and increased membranes degradability after exposure to lysozyme. Strong Vero cells adhesion was not observed in any of the tested membrane formulations. The overall results indicate that the majority of the prepared membranes meet the performance requirements of temporary nonbiodegradable burn dressings (e.g. adequate values of mechanical resistance and ductility, low values of in vitro cellular adhesion on their surfaces, low extent of degradation when exposed to lysozyme solution, and high stability in aqueous solutions).

Composite membranes of alginate and chitosan reinforced with cotton or linen fibers incorporating epidermal growth factor.

Suture threads of cotton or linen, in crossed and random orientation, were added to alginate-chitosan membranes intended to wound coatings application to improve the mechanical properties. The elongation at break increased to about 5 and 8 times for membranes with linen and cotton, respectively, both in the crossed orientation. The addition of the threads increased roughness and opacity of the membranes and reduced the liquid absorption capacity and water vapor transmission rate. The lowest toxicity to human fibroblasts was observed for extracts of membranes produced with linen, and incorporation in them of epidermal growth factor was able to slightly increase cell proliferation.

Comparison of the properties of membranes produced with alginate and chitosan from mushroom and from shrimp.

Dense and porous chitosan-alginate membranes (1:1 in mass) useful as coverages of skin wounds treated through cell therapy were produced using chitosan of different chain sizes from fungal (white mushrooms) and animal (shrimp shells) sources. Porous materials were obtained by adding the surfactant Poloxamer 188 to the formulations. The influence of chitosan type on membranes physicochemical properties and toxicity to fibroblasts was evaluated. Porosity was noticed to be more pronounced in membranes obtained with fungal chitosan and increased with its molecular mass. These formulations showed the highest values of thickness, roughness, opacity, liquid uptake and water vapor permeability. The membranes were not toxic to fibroblasts, but the lowest cytotoxicity values (0.16-0.21%) were observed for membranes prepared with fungal chitosan in the presence of surfactant. In conclusion, it is possible to replace chitosan from animal sources by chitosan of fungal origin to produce membranes with negligible cytotoxicity while maintaining appropriate physicochemical properties.

Properties of PLA/PCL particles as vehicles for oral delivery of the androgen hormone 17α-methyltestosterone.

The aim of this study was to produce PLA (poly(lactic acid)) and PCL (polycaprolactone) oral carriers through the precipitation of the polymer solutions using supercritical CO2 as an antisolvent for the controlled release of the hydrophobic model drug 17α-methyltestosterone (MT). Such drug is a steroidal hormone used orally to develop and sustain primary and secondary male sex characteristics, e.g. for female Nile tilapia sex reversal in aquaculture. The influence of hormone, PLA and PCL concentrations on particle formation was analyzed, showing that high PCL concentrations produced particles with rougher surfaces and greater mean diameters. The incorporation efficiency of MT ranged from 20 to 51%, and its addition resulted in increases in particle mean diameter from 23 to 54 µm. Aggregation was observed for particles incorporating or not MT and high concentrations of MT led to the formation of more amorphous structures, changing the thermal behavior of the particles. The exposure of the PLA/PCL particles to pH conditions simulating gastrointestinal fish conditions showed that hormone release fraction at acidic pH ranged from 8 to 63% (over 2h), while in the basic pH the proportion released varied from 23 to 60% (over 10h), reaching levels adequate for the desired in vivo activity.

Kanamycin incorporation in lipid vesicles prepared by ethanol injection designed for tuberculosis treatment.

The primary goal of this study was the production of liposomes encapsulating kanamycin for drug administration by inhalation. The selected drug is indicated for multiresistant tuberculosis, and administration through inhalation allows both local delivery of the drug to the lungs and systemic therapy. The ethanol injection method used for the liposome production is easily scaled up and is characterized by simplicity and low cost. Vesicles were prepared using different lipid compositions, including hydrogenated soybean phosphatidylcholine and cholesterol (SPC/Chol), egg phosphatidylcholine and cholesterol (EPC/Chol), distearoyl phosphatidylcholine and cholesterol (DSPC/Chol), distearoyl phosphatidylcholine, dimyristoyl phosphatidylethanolamine and cholesterol (DSPC/DMPE/Chol), dipalmitoyl phosphatidylcholine and cholesterol (DPPC/Chol) and dipalmitoyl phosphatidylcholine, dipalmitoyl phosphatidylglycerol and cholesterol (DPPC/DPPG/Chol). The effects of different operational conditions for vesicle production and drug encapsulation were evaluated, aiming at a compromise between final process cost and suitable vesicle characteristics. The best performance concerning drug incorporation was achieved with the DSPC/Chol system, although its production cost was considerably larger than that of the natural lipids formulations. Encapsulation efficiencies up to 63% and final drug to lipid molar ratios up to 0.1 were obtained for SPC/Chol vesicles presenting mean diameters of 132 nm incubated at 60 degrees C with the drug for 60 min at an initial drug-to-lipid molar ratio of 0.16.

Chitosan-alginate membranes accelerate wound healing.

The purpose of this study was to evaluate the efficacy of chitosan-alginate membrane to accelerate wound healing in experimental cutaneous wounds. Two wounds were performed in Wistar rats by punching (1.5 cm diameter), treated with membranes moistened with saline solution (CAM group) or with saline only (SL group). After 2, 7, 14, and 21 days of surgery, five rats of each group were euthanized and reepithelialization was evaluated. The wounds/scars were harvested for histological, flow cytometry, neutrophil infiltrate, and hydroxyproline analysis. CAM group presented higher inflammatory cells recruitment as compared to SL group on 2(nd) day. On the 7(th) day, CAM group showed higher CD11b(+) level and lower of neutrophils than SL group. The CAM group presented higher CD4(+) cells influx than SL group on 2(nd) day, but it decreased during the follow up and became lower on 14(th) and 21(st) days. Higher fibroplasia was noticed on days 7 and 14 as well as higher collagenesis on 21(st) in the CAM group in comparison to SL group. CAM group showed faster reepithelialization on 7(th) day than SL group, although similar in other days. In conclusion, chitosan-alginate membrane modulated the inflammatory phase, stimulated fibroplasia and collagenesis, accelerating wound healing process in rats.

Effects of different sterilization methods on the morphology, mechanical properties, and cytotoxicity of chitosan membranes used as wound dressings.

The aim of this work was to compare the effects induced by two different sterilization methods (exposure to gamma radiation or ethylene oxide) and an antiseptic technique (immersion in 70% ethanol aqueous solution) on the morphology, tensile strength, percentage of strain at break, and in vitro cytotoxicity to Vero cells on chitosan membranes designed for wound healing. Four distinct membrane compositions were evaluated, with chitosan, glycerol, and chitin used as components. Gamma radiation, in spite of being one of the most commonly employed sterilizing agents, negatively affected the morphology of membranes composed solely by chitosan as well as the percentage of strain at break of the chitosan-membranes containing glycerol on their composition. Moreover, its use affected the color of the chitosan membranes. The use of 70% ethanol aqueous solution did not change the chitosan-membrane characteristics significantly, but its use has limitations concerning the process scale up. With ethylene oxide (EtO), chitosan-membrane morphology, percentage of strain at break, and in vitro cytotoxicity to Vero cells were not significantly affected. The tensile strength of the membranes containing chitin were reduced after the treatment with ethylene oxide; however, the obtained values were comprised in the range verified for normal human skin. Therefore, because the final characteristics of the membranes treated with ethylene oxide are appropriate when considering their use as wound healing devices, and because this sterilization process is easily adjusted to use on an industrial scale, EtO can be considered the most adequate sterilizing agent for chitosan membranes. However, it should be considered that this chemical is associated with toxicity, flammability, and environmental risks, as well as with possible material contamination with ethylene oxide residues.

Analysis of cellular morphology, adhesion, and proliferation on uncoated and differently coated PVC tubes used in extracorporeal circulation (ECC).

The biggest challenge to improve extracorporeal circulation (ECC) circuits lays on avoiding platelet adhesion to their surfaces, because this contributes to thrombus formation, resulting in the activation of blood coagulation. One approach to minimize this effect is to improve the biocompatibility of ECC circuits by modifying their surfaces. This can be achieved by coating them with heparin or phospholipids. The present study investigated the adhesion and morphology characteristics of fibroblastic and blood cells cultured on uncoated poly (vinyl) chloride PVC tubes as well as on heparin, phosphatidylcholine (DMPC), and phosphatidylethanolamine (DMPE) -coated tubing. The results showed the importance of uniform coating regardless of the substance used, because the coatings cover the grooves on PVC surfaces, which favor cell adhesion. The comparison among the three different coatings showed the best biocompatibility results for the PVC tubes coated with heparin, followed by the coating with DMPE and with DMPC. For all coated tubes, cells did not spread on the PVC surfaces and, consequently, did not adhere to their surfaces, increasing the overall biocompatibility of PVC tubes. However, possible DMPE's alkylation, caused by sterilization, resulted in increased material hydrophobicity, which explains the decrease in fibroblastic adhesion. Furthermore, sterilization of DMPC-PVC improves its hydrophilic character, also decreasing adhesion. Based on these results, coating PVC with the phospholipids DMPC and DMPE seems to be a promising technique to improve the biocompatibility of PVC tubes, and is worthy of further investigation.

Control of the properties of porous chitosan-alginate membranes through the addition of different proportions of Pluronic F68.

This work addresses the development and characterization of porous chitosan-alginate based polyelectrolyte complexes, obtained by using two different proportions of the biocompatible surfactant Pluronic F68. These biomaterials are proposed for applications as biodegradable and biocompatible wound dressing and/or scaffolds. The results indicate that thickness, roughness, porosity and liquid uptake of the membranes increase with the amount of surfactant used, while their mechanical properties and stability in aqueous media decrease. Other important properties such as color and surface hydrophilicity (water contact angle) are not significantly altered or did not present a clear tendency of variation with the increase of the amount of surfactant added to the polyelectrolyte complexes, such as real density, average pore diameter, total pore volume and surface area. The prepared biomaterials were not cytotoxic to L929 cells. In conclusion, it is possible to tune the physicochemical properties of chitosan-alginate polyelectrolyte complexes, through the variation of the proportion of surfactant (Pluronic F68) added to the mixture, so as to enable the desired application of these biomaterials.

Use of chitosan membrane associated with polypropylene mesh to prevent peritoneal adhesion in rats.

The correction of wall abdominal defects often requires the use of implants such as polypropylene meshes. In spite of presenting good tissue acceptance, these biomaterials can migrate to adjacent viscera, promote enterocutaneos fistulas, tissue adherence and visceral erosions. In this work, the barrier effect of chitosan films associated with polypropylene meshes on adhesion formation experimentally induced in Wistar rats was evaluated. The animals were divided into two groups with 10 animals each. Animals in the CPP group were implanted with chitosan films associated with polypropylene meshes, whereas the ones in the PP group received only polypropylene meshes. After 8 days, the animals were submitted to euthanasia using CO(2) and a descriptive study focusing adhesion formation, visceral involvement with sutures and mesh peritonization was performed. Also, subimplanted material was collected for histopathology analysis. The results showed that the CPP group presented weak adhesions to the omentum over the stitch knots in eight animals. In all animals, the meshes were peritonized, not allowing their visualization after removing the chitosan films. In the PP group, six animals presented intestinal adhesions to the meshes and, in one of them, hepatic adhesion to the mesh was observed, besides omentum adhesion on more than 50% of the mesh area. The protective effect of chitosan films when sutured over polypropylene meshes, as well as no exacerbation of inflammation associated to the peritoneal lesions was statistically demonstrated. Therefore, chitosan films can indeed minimize the formation of peritoneal adhesions induced by polypropylene meshes in rats.