Chitosan/Hyaluronan and Alginate-Nanohydroxyapatite Biphasic Scaffold as a Promising Matrix for Osteoarthritis Disorders.

Purpose: Regenerative medicine offers new techniques for osteoarthritis (OA) disorders, especially while considering simultaneous chondral and subchondral regenerations. Methods: Chitosan and hyaluronan were chemically bound as the chondral phase and the osteogenic layer was prepared with alginate and nano-hydroxyapatite (nHAP). These scaffolds were fixed by fibrin glue as a biphasic scaffold and then examined. Results: Scanning electron microscopy (SEM) confirmed the porosity of 61.45±4.51 and 44.145±2.81 % for the subchondral and chondral layers, respectively. The composition analysis by energy dispersive X-ray (EDAX) indicated the various elements of both hydrogels. Also, their mechanical properties indicated that the highest modulus and resistance values corresponded to the biphasic hydrogel as 108.33±5.56 and 721.135±8.21 kPa, despite the same strain value as other groups. Their individual examinations demonstrated the proteoglycan synthesis of the chondral layer and also, the alkaline phosphatase (ALP) activity of the subchondral layer as 13.3±2.2 ng. After 21 days, the cells showed a mineralized surface and a polygonal phenotype, confirming their commitment to bone and cartilage tissues, respectively. Immunostaining of collagen I and II represented greater extracellular matrix (ECM) secretion in the biphasic composite group due to the paracrine effect of the two cell types on each other. Conclusion: For the first time, the ability of this biphasic scaffold to regenerate both tissue types was evaluated and the results showed satisfactory cellular commitment to bone and cartilage tissues. Thus, this scaffold can be considered a new strategy for the preparation of implants for OA.

Physical, mechanical, and biological performance of chitosan-based nanocomposite coating deposited on the polycaprolactone-based 3D printed scaffold: Potential application in bone tissue engineering.

Recently, coating on composite scaffolds has attracted many researchers' attention to improve scaffolds' properties. In this research, a 3D printed scaffold was fabricated from polycaprolactone (PCL)/magnetic mesoporous bioactive glass (MMBG)/alumina nanowire (Al2O3, Optimal percentage 5 %) (PMA) and then coated with chitosan (Cs)/multi-walled carbon nanotubes (MWCNTs) by an immersion coating method. Structural analyses such as XRD and ATR-FTIR confirmed the presence of Cs and MWCNTs in the coated scaffolds. The SEM results of the coated scaffolds showed homogeneous three-dimensional structures with interconnected pores compared to the uncoated scaffolds. The coated scaffolds exhibited an increase in compression strength (up to 16.1 MPa) and compressive modulus (up to 40.83 MPa), improved surface hydrophilicity (up to 32.69°), and decrease in degradation rate (68 % remaining weight) compared to the uncoated scaffolds. The increase in apatite formation in the scaffold coated with Cs/MWCNTs was confirmed by SEM, EDAX, and XRD tests. Coating the PMA scaffold with Cs/MWCNTs leads to the viability and proliferation of MG-63 cells and more secretion of alkaline phosphatase and Ca activity, which can be introduced as a suitable candidate for use in bone tissue engineering.

The role of three-dimensional scaffolds based on polyglycerol sebacate/ polycaprolactone/ gelatin in the presence of Nanohydroxyapatite in promoting chondrogenic differentiation of human adipose-derived mesenchymal stem cells.

BACKGROUND: Tissue engineering for cartilage regeneration has made great advances in recent years, although there are still challenges to overcome. This study aimed to evaluate the chondrogenic differentiation of human adipose-derived mesenchymal stem cells (hADSCs) on three-dimensional scaffolds based on polyglycerol sebacate (PGS) / polycaprolactone (PCL) / gelatin(Gel) in the presence of Nanohydroxyapatite (nHA). MATERIALS AND METHODS: In this study, a series of nHA-nanocomposite scaffolds were fabricated using 100:0:0, 60:40:0, and 60:20:20 weight ratios of PGS to PCL: Gel copolymers through salt leaching method. The morphology and porosity of prepared samples was characterized by SEM and EDX mapping analysis. Also, the dynamic contact angle and PBS adsorption tests are used to identify the effect of copolymerization and nanoparticles on scaffolds' hydrophilicity. The hydrolytic degradation properties were also analyzed. Furthermore, cell viability and proliferation as well as cell adhesion are evaluated to find out the biocompatibility. To determine the potential ability of nHA-nanocomposite scaffolds in chondrogenic differentiation, RT-PCR assay was performed to monitor the expression of collagen II, aggrecan, and Sox9 genes as markers of cartilage differentiation. RESULTS: The nanocomposites had an elastic modulus within a range of 0.71-1.30 MPa and 0.65-0.43 MPa, in dry and wet states, respectively. The PGS/PCL sample showed a water contact angle of 72.44 ± 2.2°, while the hydrophilicity significantly improved by adding HA nanoparticles. It was found from the hydrolytic degradation study that HA incorporation can accelerate the degradation rate compared with PGS and PGS/PCL samples. Furthermore, the in vitro biocompatibility tests showed significant cell attachment, proliferation, and viability of adipose-derived mesenchymal stem cells (ADMSCs). RT-PCR also indicated a significant increase in collagen II, aggrecan and Sox9 mRNA levels. CONCLUSIONS: Our findings demonstrated that these nanocomposite scaffolds promote the differentiation of hADSCs into chondrocytes possibly by the increase in mRNA levels of collagen II, aggrecan, and Sox9 as markers of chondrogenic differentiation. In conclusion, the addition of PCL, Gelatin, and HA into PGS is a practical approach to adjust the general features of PGS to prepare a promising scaffold for cartilage tissue engineering.

Biomimetic biphasic scaffolds in osteochondral tissue engineering: Their composition, structure and consequences.

Approaches to the design and construction of biomimetic scaffolds for osteochondral tissue, show increasing advances. Considering the limitations of this tissue in terms of repair and regeneration, there is a need to develop appropriately designed scaffolds. A combination of biodegradable polymers especially natural polymers and bioactive ceramics, shows promise in this field. Due to the complicated architecture of this tissue, biphasic and multiphasic scaffolds containing two or more different layers, could mimic the physiology and function of this tissue with a higher degree of similarity. The purpose of this review article is to discuss the approaches focused on the application of biphasic scaffolds for osteochondral tissue engineering, common methods of combining layers and the ultimate consequences of their use in patients were discussed.

Sustained release of valproic acid loaded on chitosan nanoparticles within hybrid of alginate/chitosan hydrogel with/without stem cells in regeneration of spinal cord injury.

Hydrogels have been increasingly applied in tissue regeneration and drug delivery systems (DDS). In this study, the capacity of valproic acid (Val) encapsulated within hybrid of alginate (Alg)-chitosan (Cs) (Alg-Cs) hydrogel containing Cs nanoparticle (Npch) with/without human endometrial stem cells (hEnSC) was initially examined for regeneration of spinal cord injury (SCI). To evaluate the stability of the synthesized hydrogels zeta potential necessary measurements were made. Physicochemically, the developed hydrogels were evaluated using Fourier-transform infrared (FTIR) spectroscopy. The physical properties including degradation rate, swelling ability, and tunability of the synthesized hydrogels were studied. To evaluate the nerve regeneration ability of the synthesized hydrogels, 35 Sprague-Dawley rats were undergone SCI. The spinal cords were exposed using laminectomy in T9-T10 area and the hemi-section SCI model was made. The rats were then randomly divided into 5 groups (n = 7) including, Alg-Cs/Npch, Alg-Cs/Npch/hEnSCs, Alg-Cs/Npch/Val, and Alg-Cs/Npch/hEnScs/Val, and the control groups without any intervention. The FTIR spectra showed band frequencies and assignments of Val, Alg-Cs, and alginate. Nanoparticles were formulated with a mean diameter of 187 and 210 nm, for Val/Alg-Cs and Alg-Cs, respectively. The loading of Val into Alg-Cs led to its reduced size by about 40 nm. The Cs-Npch/Val hydrogels degraded faster than the Alg-Cs-/Npch/Val hydrogel specifically in extended time of incubation. A higher swelling capacity of Alg-Cs/Npch hydrogel, compared to Cs/Npch/Val and Alg-Cs/Npch/Val hydrogels, was found. The Cs-Npch/Val hydrogels degraded faster than Alg-Cs-/Npch/Val hydrogel. The Alg-Cs/Npch/hEnSCs/Val could regenerate the damaged nerve fibers and histologically prevent the SCI-induced vacuolization spaces. The prepared Alg-Cs/Npch/Val could be a suitable polymeric carrier for taurine drugs as bioactive substrate in nerve tissue engineering (NTE) and DDS.

PCL-based 3D nanofibrous structure with well-designed morphology and enhanced specific surface area for tissue engineering application.

Tissue engineering opens a new horizon for biological tissue replacement applications. Scaffolds, appropriate cells, and signaling induction are the main three determinant parameters in any tissue engineering applications. Designing a suitable scaffold which can mimic the cellular inherent and natural habitation is of great importance for cellular growth and proliferation. Just like a natural extracellular matrix (ECM), scaffolds provide the cells with an environment for performing biological functions. Accordingly, vast surface area and three-dimensional nanofibrous structures are among the pivotal characteristics of functional scaffolds in tissue engineering, and enhancement of their properties is the main purpose of the present research. In our previous study, a patterned structure composed of continuous nanofibers and microparticles was introduced. In this work, a new modification is applied for adjustment of the surface area of an electrospun/electrosprayed scaffold. For this purpose, at predetermined stages during electrospinning/electrospraying, the nitrogen gas is flushed through the mesh holes of the collector in the opposite direction of the jet movement. This method has led to the formation of very thin nanofibrous layers at nitrogen flush intervals by providing a cooling effect of the sweeping nitrogen. As a consequence, a straticulated structure has been fabricated which possesses extremely high surface/volume ratio. The porosity, water absorption, and morphological analysis were conducted on the obtained scaffold. In vitro cytocompatibility assessments as well as histological analysis demonstrated that the fabricated scaffold provides a proper substrate for cellular attachment, proliferation and infiltration. These findings can be advantageous in three-dimensional tissue engineering such as bone tissue engineering applications. Furthermore, according to the advanced microstructure and vast surface area of the fabricated samples, they can be applied in many other applications, such as membrane, filtration, etc.

Isolation and characterization of apical papilla cells from root end of human third molar and their differentiation into cementoblast cells: an in vitro study.

BACKGROUND: Periodontal regeneration, treatment of periodontal-related diseases and improving the function of implants are global therapeutic challenges. The differentiation of human stem cells from apical papilla into cementoblasts may provide a strategy for periodontitis treatment. This study aimed to evaluate the differentiation of primary human stem cells apical papilla (hSCAPs) to cementoblast cells. MATERIAL AND METHODS: SCAPs cells were isolated from human third molar and then incubated for 21 days in a differentiation microenvironment. Alkaline phosphatase (ALP) and Alizarin red S staining assays were performed to evaluate the calcium deposition and formation of hydroxyapatite in the cultured hSCAPs microenvironment. Real-time polymerase chain reaction (RT-PCR) assay was performed for cementum protein 1 (CEMP1), collagen type I (COL1), F-Spondin (SPON1), osteocalcin (OCN), and osteopontin (OPN) as specific markers of cementoblasts and their progenitors. RESULTS: ALP phosphatase activity in day 21 of treatment demonstrated a significant increase in ALP compared to the control. Alizarin red S staining assay showed that the differentiated hSCAPs offered a great amount of calcium deposition nodules compared to the control. The increased expression level of CEMP1, OCN, OPN, COL1 and Spon1 was observed in days 7, 14 and 21 compared to the control, while greatest expression level was observed in day 21. CONCLUSION: In conclusion, the differentiation microenviroment is convenient and useful for promoting the differentiation of hSCAPs into cementoblast.

Chitosan-Placental ECM Composite Thermos-Responsive Hydrogel as a Biomimetic Wound Dressing with Angiogenic Property.

Attempts are being made to develop an ideal wound dressing with excellent biomechanical and biological properties. Here, a thermos-responsive hydrogel is fabricated using chitosan (CTS) with various concentrations (1%, 2.5%, and 5% w/v) of solubilized placental extracellular matrix (ECM) and 20% ß-glycerophosphate to optimize a smart wound dressing hydrogel with improved biological behavior. The thermo-responsive CTS (TCTS) alone or loaded with ECMs (ECM-TCTS) demonstrate uniform morphology using SEM. TCTS and ECM1%-TCTS and ECM2.5%-TCTS show a gelation time of 5 min at 37 °C, while no gel formation is observed at 4 and 25 °C. ECM5%-TCTS forms gel at both 25 and 37 °C. The degradation and swelling ratios increase as the ECM content of the hydrogel increase. All the constructs show excellent biocompatibility in vitro and in vivo, however, the hydrogels with a higher concentration of ECM demonstrate better cell adhesion for fibroblast cells and induce expression of angiogenic factors (VEGF and VEGFR) from HUVEC. Only the ECM5%-TCTS has antibacterial activity against Acinetobacter baumannii ATCC 19606. The data obtained from the current study suggest the ECM2.5%-TCTS as an optimized smart biomimetic wound dressing with improved angiogenic properties now promises to proceed with pre-clinical and clinical investigations.

An investigation of the effect of brain atrophy on brain injury in multiple sclerosis.

Multiple sclerosis (MS) is a disease of the central nervous system (CNS) that affects the brain and spinal cord. It is estimated that the average prevalence of MS is 35.9 cases per 100,000 and a total of 2.8 million people worldwide have MS. Brain atrophy is usually seen in the early stages of MS, and its progress is faster than healthy people. The present study was a numerical study that uses the Fluid-structure interaction (FSI) model to investigate the effect of brain atrophy on brain injury in MS. Firstly, a healthy model was constructed from MRI images and validated by experimental data. Then three models with different degrees of brain atrophy, which showed the rate of brain atrophy in different years in MS patients, were developed to model the brain atrophy in MS. The models were subjected to two different types of impact conditions. Type I, which only produced a translational motion and the HIC value of 744, was applied to each model. Type II produced both translational and rotational motion. In this type of impact, the experimental kinematics, with peaks of 450 g for the translational acceleration and 26.2 krad/s2 for the rotational acceleration, were applied to the nodes that located in the center of gravity of the head models and the results were extracted from each one. According to the results of impact type I, the pressure of the frontal lobe of the brain is 149,647 Pa in the health model and 137,690 Pa in the model with severe atrophy.

An investigation of cerebral bridging veins rupture due to head trauma.

Subdural hematoma (SDH) is common abnormality that is caused by the rupture of cerebral bridge veins (BVs). It occurs in more than 30% of severe head injuries. The purpose of this research was to develop a numerical model to examine the effects of brain atrophy and age on the rupture of bridging veins in subdural hematoma. Three types of models were developed to simulate subdural hematoma, namely global solid, global FSI, and local solid models. In the next step, a head impact with the head injury criterion (HIC) value of 744 was applied as a loading condition to global models. For the global solid models, we measured the relative displacement between the skull and brain. We extracted the pressure distribution from the global FSI models. The data were used as boundary conditions on the local models to evaluate the damage to the cerebral bridge veins precisely The results showed that the relative displacement was greater in the atrophied model compared to the healthy one (2.64 and 2.20 mm, respectively). In addition, the pressure value was higher in atrophied models. In the healthy local model, the maximum strain on BVs was around 1.38, while in the atrophied model, it was 2.77. The head impact, which had a HIC value of 744, did not cause serious injury to a human with a healthy brain, but it caused severe damage to an atrophied brain. The degeneration of the brain and intracranial space changes are two important factors for the movement of the brain and its vulnerability to impact.

Electro-conductive 3D printed polycaprolactone/gold nanoparticles nanocomposite scaffolds for myocardial tissue engineering.

The human's heart cannot regenerate after a wound by itself. So myocardial tissue can be damaged, leading to acute inflammation and scar. To overcome this issue, three dimensional (3D) scaffolds with appropriate properties have been proposed. In this study, Poly ε-caprolactone (PCL)/Gold nanoparticles (GNPs) nanocomposite scaffolds containing 0, 0.25 and 0.5 wt% GNPs were prepared by 3-D printing by using Fused Deposition Modeling (FDM) technique. Scanning electron microscopy (SEM), Transmission electron microscopy (TEM), Fourier transform-infrared spectroscopy (FTIR-ATR), and X-ray diffraction (XRD) were then used to characterize the scaffolds. Also, mechanical properties, electrical conductivity, contact angle, and thermal behavior of the scaffolds were measured. According to the results, the scaffold containing 0.5 wt% GNPs corroborated optimal properties including appropriate mechanical properties, adequate wettability and suitable electrical conductivity for cardiovascular application, as compressive strength and electrical conductivity were increased approximately by 9.1% and 25%, respectively. In contrast, contact angle was decreased about 38%, which caused the scaffolds' hydrophilicity. Overall the electrocunductive 3D PCL/GNPs 0.5 wt% scaffold could be developed with the control of some parameters that could be well implemented by this fabrication method; also, the addition of GNPs to improve some properties can be regarded as a promising candidate for myocardial tissue engineering.

Preparation and characterization of a new bio nanocomposites based poly(glycerol sebacic-urethane) containing nano-clay (Cloisite Na+ ) and its potential application for tissue engineering.

Nanocomposites containing clay nanoparticles often present favorable properties such as good mechanical and thermal properties. They frequently have been studied for tissue engineering (TE) and regenerative medicine applications. On the other hand, poly(glycerol sebacate) (PGS), a revolutionary bioelastomer, has exhibited substantial potential as a promising candidate for biomedical application. Here, we present a facile approach to synthesizing stiff, elastomeric nanocomposites from sodium-montmorillonite nano-clay (MMT) in the commercial name of Cloisite Na+ and poly(glycerol sebacate urethane) (PGSU). The strong physical interaction between the intercalated Cloisite Na+ platelets and PGSU chains resulted in desirable property combinations for TE application to follow. The addition of 5% MMT nano-clay resulted in an over two-fold increase in the tensile modulus, increased the onset thermal decomposition temperature of PGSU matrix by 18°C, and noticeably improved storage modulus of the prepared scaffolds, compared with pure PGSU. As well, Cloisite Na+ enhanced the hydrophilicity and water uptake ability of the samples and accelerated the in-vitro biodegradation rate. Finally, in-vitro cell viability assay using L929 mouse fibroblast cells indicated that incorporating Cloisite Na+ nanoparticles into the PGSU network could improve the cell attachment and proliferation, rendering the synthesized bioelastomers potentially suitable for TE and regenerative medicine applications.

Introducing a flexible drug delivery system based on poly(glycerol sebacate)-urethane and its nanocomposite: potential application in the prevention and treatment of oral diseases.

In this study, a novel biopolymer based on poly(glycerol sebacic)-urethane (PGS-U) and its nanocomposites containing Cloisite@30B were synthesized by facile approach in which the crosslinking was created by aliphatic hexamethylene diisocyanate (HDI) at room temperature and 80 °C. Moreover, metronidazole and tetracycline drugs were selected as target drugs and loaded into PGSU based nanocomposites. A uniform and continuous microstructure with smooth surface is observed in the case of pristine PGS-U sample. The continuity of microstructure is observed in the case of all bionanocomposites. XRD result confirmed an intercalated morphology for PGSU containing 5 wt% of clay nanoparticles with a d-spacing 3.4 nm. The increment of nanoclay content up to 5%, the ultimate tensile stress and elastic modulus were obtained nearly 0.32 and 0.83 MPa, which the latter was more than eight-fold than that of pristine PGS-U. A sustained release for both dugs was observed by 200 h. The slowest and controlled drug release rate was determined in the case of PGSU containing 5 wt% clay and cured at 80 °C. A non-Fickian diffusion can be concluded in the case of tetracycline release via PGS-U/nanoclay bionanocomposites, while a Fickian process was detected in the case of metronidazole release by PGS-U/nanoclay bionanocomposites. As a result, the designed scaffold showed high flexibility, which makes it an appropriate option for utilization in the treatment of periodontal disease.

Fabrication of Silk Scaffold Containing Simvastatin-Loaded Silk Fibroin Nanoparticles for Regenerating Bone Defects

Background: In the present study, a tissue engineered silk fibroin (SF) scaffold containing simvastatin-loaded silk fibroin nanoparticles (SFNPs) were used to stimulate the regeneration of the defected bone. Methods: At first, the porous SF scaffold was prepared using freeze-drying. Then simvastatin-loaded SFNPs were made by dissolvation method and embedded in the SF scaffold. Afterwards, the scaffold and the NPs were characterized in terms of physicochemical properties and the ability to release the simvastatin small molecule. Results: The results exhibited that the SF scaffold had a porous structure suitable for releasing the small molecule and inducing the proliferation and attachment of osteoblast cells. SFNPs containing simvastatin had spherical morphology and were 174 ± 4 nm in size with -24.5 zeta potential. Simvastatin was also successfully encapsulated within the SFNPs with 68% encapsulation efficiency. Moreover, the small molecule revealed a sustained release profile from the NPs during 35 days. The results obtained from the in vitro cell-based studies indicated that simvastatin-loaded SFNPs embedded in the scaffold had acceptable capacity to promote the proliferation and alkaline phosphatase production of osteoblast cells while inducing osteogenic matrix precipitation. Conclusion: The SF scaffold containing simvastatin-loaded SFNPs could have a good potential to be used as a bone tissue-engineered construct.

Review of Bioprinting in Regenerative Medicine: Naturally Derived Bioinks and Stem Cells.

Regenerative medicine offers the potential to repair or substitute defective tissues by constructing active tissues to address the scarcity and demands for transplantation. The method of forming 3D constructs made up of biomaterials, cells, and biomolecules is called bioprinting. Bioprinting of stem cells provides the ability to reliably recreate tissues, organs, and microenvironments to be used in regenerative medicine. 3D bioprinting is a technique that uses several biomaterials and cells to tailor a structure with clinically relevant geometries and sizes. This technique's promise is demonstrated by 3D bioprinted tissues, including skin, bone, cartilage, and cardiovascular, corneal, hepatic, and adipose tissues. Several bioprinting methods have been combined with stem cells to effectively produce tissue models, including adult stem cells, embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), and differentiation techniques. In this review, technological challenges of printed stem cells using prevalent naturally derived bioinks (e.g., carbohydrate polymers and protein-based polymers, peptides, and decellularized extracellular matrix), recent advancements, leading companies, and clinical trials in the field of 3D bioprinting are delineated.

Conductive Biomaterials as Substrates for Neural Stem Cells Differentiation towards Neuronal Lineage Cells.

The injuries and defects in the central nervous system are the causes of disability and death of an affected person. As of now, there are no clinically available methods to enhance neural structural regeneration and functional recovery of nerve injuries. Recently, some experimental studies claimed that the injuries in brain can be repaired by progenitor or neural stem cells located in the neurogenic sites of adult mammalian brain. Various attempts have been made to construct biomimetic physiological microenvironment for neural stem cells to control their ultimate fate. Conductive materials have been considered as one the best choices for nerve regeneration due to the capacity to mimic the microenvironment of stem cells and regulate the alignment, growth, and differentiation of neural stem cells. The review highlights the use of conductive biomaterials, e.g., polypyrrole, polyaniline, poly(3,4-ethylenedioxythiophene), multi-walled carbon nanotubes, single-wall carbon nanotubes, graphene, and graphite oxide, for controlling the neural stem cells activities in terms of proliferation and neuronal differentiation. The effects of conductive biomaterials in axon elongation and synapse formation for optimal repair of central nervous system injuries are also discussed.

Nanocontainers for drug delivery systems: A review of Halloysite nanotubes and their properties.

Halloysite nanotubes (HNTs) are known as inexpensive and available nanomaterials that are rich in functionality, environmentally benign, and also safe and easy to process. As well, good particle size (i.e. nanoscale) and perfect tubular microstructures of these materials make them to be used extensively as drug carriers. Also, the unique physical and chemical properties of their internal and external surfaces are the greatest priority for the drug encapsulation controlling and releasing. In this review, is tried to emphasis on the main properties of HNTs to manage and develop effective drug delivery tools in the biomedical and pharmaceutical fields.