Tissue engineering application combining epoxidized natural rubber blend and mesenchymal stem cells in in vivo response.

This study aimed to investigate biocompatibility, integration, and tissue host response of the Poly (Lactic-co-Glycolic acid) (PLGA)/Poly (isoprene) (PI) epoxidized (PLGA/PIepox) innovative scaffold combined with adipose derived mesenchymal stem cells (ADSC). We implanted the scaffold subcutaneously on the back of 18 female rats and monitored them for up to 14 days. When compared to controls, PLGA/PIepox + ADSC demonstrated an earlier vascularization, a tendency of inflammation reduction, an adequate tissue integration, higher cell proliferation, and a tendency of expression of collagen decreasing. However, 14 days post-implantation we found similar levels of CD31, Ki67 and AE1/AE3 in PLGA/PIepox when compared to control groups. The interesting results, lead us to the assumption that PLGA/PIepox is able to provide an effective delivery system for ADSC on tissue host. This animal study assesses PLGA/PIepox + ADSC in in vivo tissue functionality and validation of use, serving as a proof of concept for future clinical translation as it presents an innovative and promising tissue engineering opportunity for the use in tissue reconstruction.

Mechanical properties, in vitro and in vivo biocompatibility analysis of pure iron porous implant produced by metal injection molding: A new eco-friendly feedstock from natural rubber (Hevea brasiliensis).

Metal injection molding (MIM) has become an important manufacturing technology for biodegradable medical devices. As a biodegradable metal, pure iron is a promising biomaterial due to its mechanical properties and biocompatibility. In light of this, we performed the first study that manufactured and evaluated the in vitro and in vivo biocompatibility of samples of iron porous implants produced by MIM with a new eco-friendly feedstock from natural rubber (Hevea brasiliensis), a promisor binder that provides elastic property in the green parts. The iron samples were submitted to tests to determine density, microhardness, hardness, yield strength, and stretching. The biocompatibility of the samples was studied in vitro with adipose-derived mesenchymal stromal cells (ADSCs) and erythrocytes, and in vivo on a preclinical model with Wistar rats, testing the iron samples after subcutaneous implant. Results showed that the manufactured samples have adequate physical, and mechanical characteristics to biomedical devices and they are cytocompatible with ADSCs, hemocompatible and biocompatible with Wistars rats. Therefore, pure iron produced by MIM can be considered a promising material for biomedical applications.

In situ drug release measuring in α-TCP cement by electrochemical impedance spectroscopy.

The use of drug delivery systems is a good technique to leave the right quantity of medicine in the patient's body in a suitable dose, because the drug application is delivered directly to the affected region. The current techniques such as HPLC and UV-Vis for the drug delivery calculation has some disadvantages, as the accuracy and the loss of the sample after characterization. With the aim of reducing the amount of material used during the characterization and have a non-destructive test with instantaneous results, the present paper shows the possibility of using electrochemical impedance spectroscopy (EIS) to have a drug delivery measurement during the release phenomena for a calcium phosphate cement (CFC) delivery system with gentamicin sulfate (GS) and lidocaine hydrochloride (LH), at a ratio of 1% and 2%, respectively. The equivalent circuit and the chemical mechanism involved during the measurements have been proposed as a tool to determine the drug delivery profile. The method has been compared with the UV-Vis technique. XRD was realized to verify conditions, before and after release. It was possible to verify the potential for using EIS as an instant technique to quantify drug delivery.

Tissue-engineered solution containing cells and biomaterials-an in vitro study: A perspective as a novel therapeutic application.

Some biomaterial scaffolds can positively interfere with tissue regeneration and are being developed to successfully repair the tissue function. The possibility of using epithelial cells combined with biomaterials appears to be a new option as therapeutic application. This combination emerges as a possibility for patients with Mayer-Rokitansky-Kuster-Hauser syndrome which requires vaginal repair and can be performed with tissue-engineered solution containing cells and biomaterials. It is expected that tissue-engineered solution containing cells and biomaterials would promote tissue repair in a more efficient, modern, and safe way. This study tested the efficiency of tissue-engineered solution containing human malignant melanoma cell line (HMV-II) and different biomaterials, including Cellprene®, Membracel®, and poly lactic-co-glycolic acid/epoxidized polyisoprene. The cells adhered better on poly lactic-co-glycolic acid/epoxidized polyisoprene, and it was found that tissue-engineered solution may also contain mesenchymal stem cells cultivated on poly lactic-co-glycolic acid/epoxidized polyisoprene. Histological, immunofluorescence, and scanning electron microscopy analyses were performed. These initial in vitro results suggest that tissue-engineered solution containing cells and poly lactic-co-glycolic acid/epoxidized polyisoprene is a potential for tissue reconstruction.

Polydimethylsiloxane/nano calcium phosphate composite tracheal stents: Mechanical and physiological properties.

In this study, we report the production and characterization of tracheal stents composed of polydimethylsiloxane/nanostructured calcium phosphate composites obtained by reactive synthesis. Tracheal stents were produced by transfer molding, and in vivo tests were carried out. PDMS was combined with H3 PO4 and Ca(OH)2 via an in situ reaction to obtain nanoparticles of calcium phosphate dispersed within the polymeric matrix. The incorporation of bioactive inorganic substances, such as calcium phosphates, improved biological properties, and the in situ reaction allowed tight coupling of particles to the matrix. Results showed the presence of the nanoparticles of DCPA and CDHA. The porosity generated during mixing decreased the tensile strength and tear properties. Composites presented higher values of cell viability compared with those for PDMS. In vivo tests indicated the presence of inflammatory tissue 30 days after implantation in both cases. Thus, the present biomaterial shows potential for application in tracheal disease, however further evaluation is needed. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2018. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 545-553, 2019.

Fiber-reinforced silicone for tracheobronchial stents: An experimental study.

A trachea is a tubular structure composed of smooth muscle that is reinforced with cartilage rings. Some diseases can cause sagging in smooth muscle and cartilaginous tissue. The end result is reduction (narrowing) of the trachea diameter. A solution to this problem is the use of tracheal stents, which are small tubular devices made of silicone. One is inserted into the trachea to prevent or correct its constriction. The purpose of tracheal stent use is to maintain cartilage support that would otherwise be lost in the airway. Current tracheal stent models present limitations in terms of shape and characteristics of the silicone used in their production. One of the most important is the large thickness of the wall, which makes its placement difficult; this mainly applies to pediatric patients. The wall thickness of the stent is closely related to the mechanical properties of the material. This study aims to test the reinforcement of silicone with three kinds of fibers, and then stents that were produced using fiber with the best compressive strength characteristics. Silicone samples were reinforced with polypropylene (PP), polyamide (PA), and carbon fiber (CF) at concentrations of 2% and 4% (vol%), which then underwent tensile strength and Shore A hardness testing. Samples with fiber showed good characteristics; surface analyses were carried out and they were used to produce stents with an internal diameter of 11 or 13mm and a length of 50mm. Stents underwent compression tests for qualitative evaluation. Samples with 2% and 4% CF blends showed the best mechanical performance, and they were used to produce stents. These samples presented similar compressive strengths at low deformation, but stents with a 4% CF blend exhibited improved compressive strength at deformations greater than 30-50% of their diameter (P ≤ 0.05). The addition of 2% and 4% CF blends conferred greater mechanical strength and resistance to the silicone matrix. This is particularly true at low deformation, which is the condition where the stent is used when implanted. In the finite element compression strength tests, the stent composite showed greater compression strength with the addition of fiber, and the results were in accordance with mechanical compression tests performed on the stents. In vivo tests showed that, after 30 days of post-implantation in sheep trachea, an inflammatory process occurred in the region of the trachea in contact with the stent composite and with the stent without fiber (WF). This response is a common process during the first few days of implantation.

Methacrylate-based root canal sealer containing chlorexidine and α-tricalcium phosphate.

The aim of this study was to develop and to characterize a methacrylate-based root canal sealer containing chlorhexidine (CHX) and α-tricalcium phosphate (α-TCP). Experimental dual cure methacrylate-based sealer was produced containing 0, 2.5, or 5 wt% of CHX and 0, 25, or 50 wt% of α-TCP. Experimental sealers were evaluated based on flow, film thickness, radiopacity, degree of conversion (DC), degradation in water, pH and antibacterial activity. Flow ranged from 15.09 ± 0.11 to 17.47 ± 0.42 mm. All groups presented mean film thickness lower than 50 µm and had radiopacity equivalent to 3 mmAl. DC was higher than 60% for all compositions. The weight loss (WL) ranged 0.12-3.47%. The groups containing 5% of CHX presented the highest WL and the lower pH values after 28 days of water immersion. All chlorexidine-compositions exhibited antibacterial efficacy against Enterococcus faecalis on direct contact and agar diffusion tests. CHX and α-TCP addition at an experimental methacrylate-based root canal sealer influenced the physicochemical properties and provided antibacterial properties. The incorporation of CHX and α-TCP could be an alternative to antibacterial sealers with potential to improve periapical healing in endodontic treatments. © 2017 Wiley Periodicals, Inc. J Biomater Res Part B: 106B: 1439-1443, 2018.

Different post-processing conditions for 3D bioprinted α-tricalcium phosphate scaffolds.

The development of 3D printing hardware, software and materials has enabled the production of bone substitute scaffolds for tissue engineering. Calcium phosphates cements, such as those based on α-tricalcium phosphate (α-TCP), have recognized properties of osteoinductivity, osteoconductivity and resorbability and can be used to 3D print scaffolds to support and induce tissue formation and be replaced by natural bone. At present, however, the mechanical properties found for 3D printed bone scaffolds are only satisfactory for non-load bearing applications. This study varied the post-processing conditions of the 3D powder printing process of α-TCP cement scaffolds by either immersing the parts into binder, Ringer's solution or phosphoric acid, or by sintering in temperatures ranging from 800 to 1500 °C. The porosity, composition (phase changes), morphology, shrinkage and compressive strength were evaluated. The mechanical strength of the post-processed 3D printed scaffolds increased compared to the green parts and was in the range of the trabecular bone. Although the mechanical properties achieved are still low, the high porosity presented by the scaffolds can potentially result in greater bone ingrowth. The phases present in the scaffolds after the post-processing treatments were calcium-deficient hydroxyapatite, brushite, monetite, and unreacted α-TCP. Due to their chemical composition, the 3D printed scaffolds are expected to be resorbable, osteoinductive, and osteoconductive.

Dimensional evaluation of patient-specific 3D printing using calcium phosphate cement for craniofacial bone reconstruction.

The 3D printing process is highlighted nowadays as a possibility to generate individual parts with complex geometries. Moreover, the development of 3D printing hardware, software and parameters permits the manufacture of parts that can be not only used as prototypes, but are also made from materials that are suitable for implantation. In this way, this study investigates the process involved in the production of patient-specific craniofacial implants using calcium phosphate cement, and its dimensional accuracy. The implants were previously generated in a computer-aided design environment based on the patient's tomographic data. The fabrication of the implants was carried out in a commercial 3D powder printing system using alfa-tricalcium phosphate powder and an aqueous solution of Na2HPO4 as a binder. The fit of the 3D printed implants was measured by three-dimensional laser scanning and by checking the right adjustment to the patient's anatomical biomodel. The printed parts presented a good degree of fitting and accuracy.

Development and testing of an absorbable spring for cranial expansion in rabbits.

OBJECTIVE: Use of metal springs for treatment of craniosynostosis is gaining ground in the surgical armamentarium, as these springs simplify operative technique, help to avoid extended approaches, and thus minimize morbidity. Nevertheless, these devices have to be removed eventually. The purpose of this study was to perform cranial expansion with a fully integrated, biodegradable polymer spring in an animal model and to assess the efficacy of and histological reaction to this device. MATERIAL AND METHODS: This was an experimental, unblinded, prospective study. Twelve female New Zealand rabbits (Oryctolagus cuniculus) aged 6 weeks were randomly allocated to two groups. Control animals underwent linear craniectomy alone. Intervention animals underwent craniectomy with placement of a poly(lactic-co-glycolic acid)/polyisoprene (PLGA/PI) copolymer blend spring for cranial expansion transverse to the ostectomy. Expansion was measured radiographically over 12 weeks with amalgam markers. At the end of the experiment period, histological analysis was performed to quantify inflammatory reaction. RESULTS: The copolymer blend springs had a mean strength of 4.2N. In the intervention group, cranial expansion at the frontal markers was 9.6-11.67 mm (significantly greater than in controls). Histological analysis showed minor inflammatory reactions. CONCLUSION: In this animal model, cranial expansion by linear craniectomy followed by bioabsorbable spring placement was feasible and well tolerated by adjacent tissues.

α-Tricalcium phosphate cements modified with ß-dicalcium silicate and tricalcium aluminate: physicochemical characterization, in vitro bioactivity and cytotoxicity.

Biocompatibility, injectability and in situ self-setting are characteristics of calcium phosphate cements which make them promising materials for a wide range of clinical applications in traumatology and maxillo-facial surgery. One of the main disadvantages is their relatively low strength which restricts their use to nonload-bearing applications. α-Tricalcium phosphate (α-C3P) cement sets into calcium-deficient hydroxyapatite (CDHA), which is biocompatible and plays an essential role in the formation, growth and maintenance of tissue-biomaterial interface. ß-Dicalcium silicate (ß-C2S) and tricalcium aluminate (C3A) are Portland cement components, these compounds react with water to form hydrated phases that enhance mechanical strength of the end products. In this study, setting time, compressive strength (CS) and in vitro bioactivity and biocompatibility were evaluated to determine the influence of addition of ß-C2S and C3A to α-C3P-based cement. X-ray diffraction and scanning electron microscopy were used to investigate phase composition and morphological changes in cement samples. Addition of C3A resulted in cements having suitable setting times, but low CS, only partial conversion into CDHA and cytotoxicity. However, addition of ß-C2S delayed the setting times but promoted total conversion into CDHA by soaking in simulated body fluid and strengthened the set cement over the limit strength of cancellous bone. The best properties were obtained for cement added with 10 wt % of ß-C2S, which showed in vitro bioactivity and cytocompatibility, making it a suitable candidate as bone substitute.

ß-Dicalcium silicate-based cement: synthesis, characterization and in vitro bioactivity and biocompatibility studies.

ß-dicalcium silicate (ß-Ca2 SiO4, ß-C2 S) is one of the main constituents in Portland cement clinker and many refractory materials, itself is a hydraulic cement that reacts with water or aqueous solution at room/body temperature to form a hydrated phase (C-S-H), which provides mechanical strength to the end product. In the present investigation, ß-C2 S was synthesized by sol-gel process and it was used as powder to cement preparation, named CSiC. In vitro bioactivity and biocompatibility studies were assessed by soaking the cement samples in simulated body fluid solutions and human osteoblast cell cultures for various time periods, respectively. The results showed that the sol-gel process is an available synthesis method in order to obtain a pure powder of ß-C2 S at relatively low temperatures without chemical stabilizers. A bone-like apatite layer covered the material surface after soaking in SBF and its compressive strength (CSiC cement) was comparable with that of the human trabecular bone. The extracts of this cement were not cytotoxic and the cell growth and relative cell viability were comparable to negative control.

Development of dual-setting calcium phosphate cement using absorbable polymer.

Calcium phosphate cements used as bone substitutes generally have low mechanical strength compared with the bones of the human body. To solve these needs, we have incorporated hydrogels in the manufacture of samples made of alpha-tricalcium phosphate (α-TCP) cement, developing a system of dual-setting cement. This study aimed to produce composite materials by combining α-TCP powder and hydrogels. The composites were prepared using the synthesized powder and four different formulations of hydrogels, using either poly(N-vinyl-2-pyrrolidone) or poly(N-vinyl-2-pyrrolidone-co-acrylic acid), with either azobisisobutyronitrile or ammonium persulfate as initiator. The properties of all composites were evaluated through measuring compressive strength and apparent density and through X-ray diffraction and scanning electron microscopy. The composites showed compressive strengths of around 24 MPa. Soaking the samples in simulated body fluid formed a layer of hydroxyapatite-like crystals on the surface of some samples, showing the bioactivity of the newly developed cements and their potential use as biomaterial.

Calcium phosphate cement scaffolds with PLGA fibers.

The use of calcium phosphate-based biomaterials has revolutionized current orthopedics and dentistry in repairing damaged parts of the skeletal system. Among those biomaterials, the cement made of hydraulic grip calcium phosphate has attracted great interest due to its biocompatibility and hardening "in situ". However, these cements have low mechanical strength compared with the bones of the human body. In the present work, we have studied the attainment of calcium phosphate cement powders and their addition to poly (co-glycolide) (PLGA) fibers to increase mechanical properties of those cements. We have used a new method that obtains fibers by dripping different reagents. PLGA fibers were frozen after lyophilized. With this new method, which was patented, it was possible to obtain fibers and reinforcing matrix which furthered the increase of mechanical properties, thus allowing the attainment of more resistant materials. The obtained materials were used in the construction of composites and scaffolds for tissue growth, keeping a higher mechanical integrity.

Bioactive composite bone cement based on α-tricalcium phosphate/tricalcium silicate.

Silicon compounds are known as bioactive materials that are able to bond to the living bone tissue by inducing an osteogenic response through the stimulation and activation of osteoblasts. To improve the bioactive and mechanical properties of an α-Ca(3)PO(4)-based cement, the effects of the addition of Ca(3 SiO(5) (C(3)S) on physical, chemical, mechanical, and biological properties after soaking in simulated body fluid (SBF) were studied. The morphological and structural changes of the material during immersion were analyzed by X-ray diffraction and scanning electron microscopy. The results showed that it is possible to increase the compressive strength of the cement by adding 5% of C(3)S. Higher C(3)S contents enhance bioactivity and biocompatibility by the formation of a dense and homogeneous hydroxyapatite layer within 7 days; however, compressive strength decreases drastically as a consequence of delayed hydrolysis of α-Ca(3)(PO(4) (2). An increment in setting times and degradation rate of composites containing C(3)S was also observed.

Chemical synthesis and in vitro biocompatibility tests of poly (L-lactic acid).

Polylactic acid is a polymer of great technological interest, whose excellent mechanical properties, thermal plasticity, and bioresorbability render it potentially useful for environmental applications, as a biodegradable plastic and as a biocompatible material in biomedicine. This article discusses the synthesis and characterization of poly-L-lactic acid, obtained through two synthetic routes: direct polycondensation reactions without organic solvents, and in a supercritical medium. Tin complexes were used as catalysts in both polymerization reactions. The polymers were characterized by (1)HNMR, IR, GPC, DSC, and TGA techniques. In vitro biocompatibility tests were performed with human alveolar bone osteoblasts and there were assessed cell adhesion, proliferation and viability. The poly condensation reaction proved to be an excellent synthetic route to produce PLA polymers with different molar mass. The formation of polymers from lactic acid monomer was confirmed through techniques utilized. It was observed that cell adhesion and viability was not disturbed by the presence of the polymer, although the proliferation rate was decreased when compared to control.

Dual-setting calcium phosphate cement modified with ammonium polyacrylate.

alpha-Tricalcium phosphate bone cement, as formerly designed and developed by Driessens et al., consists of a powder composed by alpha-tricalcium phosphate (alpha-TCP) and hydroxyapatite (HA) seeds, and an aqueous solution of Na2HPO4 as mixing liquid. After mixing powder and liquid, alpha-TCP dissolves into the liquid and calcium deficient hydroxyapatite (CDHA), more insoluble than the former, precipitates as an entanglement of crystals, which causes the setting and hardening of the cement. alpha-TCP bone cement offers several advantages in comparison to calcium phosphate bioceramics and acrylic bone cements as bone graft and repairing material, like perfect adaptability to the defect size and shape, osteotransductibility, and absence of thermal effect during setting. The main handicap is its low mechanical strength. Therefore, approaching its mechanical strength to that of human bone could considerably extend its applications. In the present work, an in situ polymerization system based on acrylamide (AA) and ammonium polyacrylate (PA) as liquid reducer was added to alpha-TCP cement to increase its mechanical strength. The results showed that the addition of 20 wt% of acrylamide and 1 wt% AP to the liquid increased the compressive and tensile strength of alpha-TCP bone cement by 149 and 69% (55 and 21 MPa), respectively. The improvement in mechanical strength seems to be caused by a decrease of porosity and the reinforcing effect of a polyacrylamide network coexisting with the entanglement of CDHA crystals. The studied additives do not affect the nature of the final product of the setting reaction, CDHA, but promote the reduction of its crystal size.

Fiber-enriched double-setting calcium phosphate bone cement.

Calcium phosphate bone cements are useful in orthopedics and traumatology, their main advantages being their biocompatibility and bioactivity, which render bone tissue osteoconductive, providing in situ hardening and easy handling. However, their low mechanical strength, which, in the best of cases, is equal to the trabecular bone, and their very low toughness are disadvantages. Calcium phosphate cement compositions with mechanical properties more closely resembling those of human bone would broaden the range of applications, which is currently limited to sites subjected to low loads. This study investigated the influence of added polypropylene, nylon, and carbon fibers on the mechanical properties of double setting alpha-tricalcium phosphate-based cement, using calcium phosphate cement added to an in situ polymerizable acrylamide-based system recently developed by the authors. Although the addition of fibers was found to reduce the compression strength of the double-setting calcium phosphate cement because of increased porosity, it strongly increased the cement's toughness (J(IC)) and tensile strength. The composites developed in this work, therefore, have a potential application in shapes subjected to flexure.