Electrospun zein nanofibers loaded with curcumin as a wound dressing: enhancing properties with PSS and PDADMAC layers.

Development of wound dressings with enhanced therapeutic properties is of great interest in the modern healthcare. In this study, a zein-based nanofibrous wound dressing containing curcumin as a therapeutic agent was fabricated through electrospinning technique. In order to achieve desirable properties, such as antibacterial characteristics, reduced contact angle, and enhanced mechanical properties, the layer-by-layer technique was used for coating the surfaces of drug-loaded nanofibers by sequentially incorporating poly (sodium 4-styrene sulfonate) as a polyanion and poly (diallyldimethylammonium chloride) (PDADMAC) as a polycation. Various analyses, including scanning electron microscopy, Fourier transform infrared spectroscopy, drug release assessment., and mechanical tests were employed to assess the characteristics of the prepared wound dressings. Based on the results, coating with polyelectrolytes enhanced the Young's modulus and tensile strength of the electrospun mat from 1.34 MPa and 4.21 MPa to 1.88 MPa and 8.83 MPa, respectively. The coating also improved the controlled release of curcumin and antioxidant activity, while the outer layer, PDADMAC, exhibited antibacterial properties. The cell viability tests proved the appropriate biocompatibility of the prepared wound dressings. Moreover, our findings show that incorporation of the coating layers enhances cell migration and provides a favorable surface for cell attachment. According to the findings of this study, the fabricated nanofibrous wound dressing can be considered a promising and effective therapeutic intervention for wound management, facilitating the healing process.

Enhanced spinal cord regeneration by gelatin/alginate hydrogel scaffolds containing human endometrial stem cells and curcumin-loaded PLGA nanoparticles in rat.

Spinal cord injury (SCI) is a serious problem with a high prevalence worldwide. The weak capability of the spinal cord for regeneration in association with upregulation of inflammatory factors is two key obstacles against a full SCI repair. Curcumin is a natural substance with anti-inflammatory and neuroprotective effects. Here, we have used a combined strategy using stem cells and hybrid hydrogel scaffolds loaded with curcumin for SCI repair. Curcumin-loaded PLGA nanoparticles were prepared, characterized, and encapsulated into gelatin/alginate hydrogel scaffolds, which were then seeded by human endometrial stem cells (hEnSCs). The resulting construct was studied using in vitro and in vivo experiments on rat models. DLS, SEM, Zeta potential, and FTIR data confirmed the successful addition of curcumin to PLGA nanoparticles. SEM analyses indicated the successful addition of curcumin-loaded nanoparticles into the gelatin/alginate scaffold, as well as the adherence of the seeded EnSCs. Based on the results, the prepared constructs not only allowed the controlled release of curcumin but also could support the survival and growth of hEnSCs. Based on the results of BBB and histological experiments, the highest BBB score was related to the combined strategy, consistent with histological outcomes, in which our hEnSC-seeded gelatin/alginate scaffold containing curcumin-loaded nanoparticles led to improved structures of the white and gray matters in the SCI site, being indicative of the superior nerve fiber regeneration, compared to other studied groups. These results indicate the efficiency of the proposed method for SCI repair and broaden the scope for subsequent studies on spinal cord regeneration.

Nonlinear viscoelastic properties of human dentin under uniaxial tension.

OBJECTIVE: Dentin is a viscoelastic tissue that contributes to the load distribution in human teeth and leads to their fracture resistance. Despite previous researches on the time-dependent behavior of dentin, it is not very clear whether the viscoelastic behavior of this tissue is linear or nonlinear, and what viscoelastic constitutive equations mechanically characterize it. Therefore, the aim of this study was to describe the viscoelastic behavior of human dentin and determine the best-fitting viscoelastic model for this tissue. METHODS: After preparation of human dentin specimens from 50 subjects, tensile stress relaxation tests were performed at 1%, 3%, 5% and 7% strain amplitudes. We first evaluated the viscoelastic linearity of this tissue and then fitted the experimental data using different constitutive models, namely, 2-, 3- and 4-term Prony series for linear viscoelasticity, Fung's quasilinear viscoelastic model, and also Schapery and modified superposition models for nonlinear viscoelasticity. RESULTS: Despite an almost linear trend at small strains up to 5%, the relaxation rate generally depended on strain amplitude, indicating some degree of nonlinearity in dentine viscoelasticity. According to the results of data fitting using different models, the modified superposition formulation could best capture the viscoelastic behavior of human dentin. SIGNIFICANCE: In this study, we have quantitatively examined the viscoelastic behavior of human dentin, using a large number of samples. We have obtained the coefficients of various viscoelastic formulations, which can be utilized in subsequent researches on human dentin assuming linear, quasilinear or nonlinear viscoelasticity for this tissue.

In-vivo characterization of a 3D hybrid scaffold based on PCL/decellularized aorta for tracheal tissue engineering.

INTRODUCTION: As common treatments for long tracheal stenosis are associated with several limitations, tracheal tissue engineering is considered as an alternative treatment. AIM OF STUDY: This study aimed at preparing a hybrid scaffold, based on biologic and synthetic materials for tracheal tissue engineering. MATERIALS AND METHODS: Three electrospun polycaprolactone (PCL) scaffolds, namely E1 (pure PCL), E2 (collagen-coated PCL) and E3 (PCL blended with collagen) were prepared. Allogeneic aorta was harvested and decellularized. A biodegradable PCL stent was fabricated and inserted into the aorta to prevent its collapse. RESULT: Scaffold characterization results revealed that the 2-h swelling ratio of E2 was significantly higher than those of E1 and E3. In the first 3months, E2 and E3 exhibited almost equal degradabilities (significantly higher than that of E1). Moreover, tensile strengths of all samples were comparable with those of human trachea. Using rabbit's adipose-derived mesenchymal stem cells (AMSCs) and primary chondrocytes, E3 exhibited the highest levels of GAG release within 21days as well as collagen II and aggrecan expression. Fot the next step, AMSC-chondrocyte co-culture seeded scaffold was sutured to the acellular aorta, implanted into rabbits' muscle, and finally harvested after 4weeks of follow up. CONCLUSION: Harvested structures were totally viable due to the angiogenesis created by the muscle. H&E and alcian blue staining results revealed the presence of chondrocytes in the structure and GAG in the produced extracellular matrix. Since tracheal replacement using biologic and synthetic scaffolds usually results in tracheal collapse or granulation formation, a hybrid construct may provide the required rigidity and biocompatibility for the substitute.

Investigation of the Mechanical Properties of the Human Tracheal Cartilage.

BACKGROUND: The tracheal cartilage plays an important role in maintaining the mechanical stability of the trachea, as it keeps the trachea open and prevents its collapse under the negative pressures of the respiratory cycle. This study aimed to evaluate and compare the mechanical properties of cartilage specimens from the cranial and caudal regions of the human trachea and compare the results with respect to age and sex of the subjects. MATERIALS AND METHODS: After obtaining human trachea samples from brain-dead, organ-donating patients and storing them in appropriate conditions, the prepared cartilage samples from the cranial and caudal regions of the trachea were subjected to uniaxial tension and stress relaxation experiments to obtain the corresponding Young's modulus and relaxation percentage values, respectively. The results were compared in terms of the position (cranial or caudal) in the trachea, and age and sex of the patients. RESULTS: Based on the results, no statistically significant effect of the position in the trachea on the Young's modulus of the human tracheal cartilage samples was observed, despite the generally stiffer behavior of cartilage samples from the cranial region compared to those from the caudal region of the trachea. For both the cranial and caudal regions, no significant effect of sex on the stiffness of the tracheal cartilage was observed; further, the cartilage samples of the human trachea (from both cranial and caudal regions) of the old subjects were significantly stiffer than those of the young subjects. Based on the stress relaxation data, no significant effect of age, sex, or position on the relaxation percentage was observed. CONCLUSION: The tracheal cartilage samples of the old patients are significantly stiffer than those of the young patients. Sex and position in the trachea (cranial vs caudal) do not significantly influence the mechanical properties of the human tracheal cartilage samples. The results of this study can be useful in designing tracheal tissue-engineered scaffolds, which should be mechanically compatible with the native trachea.

Viscoelastic Properties of Human Tracheal Tissues.

The physiological performance of trachea is highly dependent on its mechanical behavior, and therefore, the mechanical properties of its components. Mechanical characterization of trachea is key to succeed in new treatments such as tissue engineering, which requires the utilization of scaffolds which are mechanically compatible with the native human trachea. In this study, after isolating human trachea samples from brain-dead cases and proper storage, we assessed the viscoelastic properties of tracheal cartilage, smooth muscle, and connective tissue based on stress relaxation tests (at 5% and 10% strains for cartilage and 20%, 30%, and 40% for smooth muscle and connective tissue). After investigation of viscoelastic linearity, constitutive models including Prony series for linear viscoelasticity and quasi-linear viscoelastic, modified superposition, and Schapery models for nonlinear viscoelasticity were fitted to the experimental data to find the best model for each tissue. We also investigated the effect of age on the viscoelastic behavior of tracheal tissues. Based on the results, all three tissues exhibited a (nonsignificant) decrease in relaxation rate with increasing the strain, indicating viscoelastic nonlinearity which was most evident for cartilage and with the least effect for connective tissue. The three-term Prony model was selected for describing the linear viscoelasticity. Among different models, the modified superposition model was best able to capture the relaxation behavior of the three tracheal components. We observed a general (but not significant) stiffening of tracheal cartilage and connective tissue with aging. No change in the stress relaxation percentage with aging was observed. The results of this study may be useful in the design and fabrication of tracheal tissue engineering scaffolds.

Mechanical Characterization and Constitutive Modeling of Human Trachea: Age and Gender Dependency.

Tracheal disorders can usually reduce the free lumen diameter or wall stiffness, and hence limit airflow. Trachea tissue engineering seems a promising treatment for such disorders. The required mechanical compatibility of the prepared scaffold with native trachea necessitates investigation of the mechanical behavior of the human trachea. This study aimed at mechanical characterization of human tracheas and comparing the results based on age and gender. After isolating 30 human tracheas, samples of tracheal cartilage, smooth muscle, and connective tissue were subjected to uniaxial tension to obtain force-displacement curves and calculate stress-stretch data. Among several models, the Yeoh and Mooney-Rivlin hyperelastic functions were best able to describe hyperelastic behavior of all three tracheal components. The mean value of the elastic modulus of human tracheal cartilage was calculated to be 16.92 ± 8.76 MPa. An overall tracheal stiffening with age was observed, with the most considerable difference in the case of cartilage. Consistently, we noticed some histological alterations in cartilage and connective tissue with aging, which may play a role in age-related tracheal stiffening. No considerable effect of gender on the mechanical behavior of tracheal components was observed. The results of this study can be applied in the design and fabrication of trachea tissue engineering scaffolds.

Regulation of Endothelial Cell Adherence and Elastic Modulus by Substrate Stiffness.

Although substrate stiffness has been previously reported to affect various cellular aspects, such as morphology, migration, viability, growth, and cytoskeletal structure, its influence on cell adherence has not been well examined. Here, we prepared three soft, medium, and hard polyacrylamide (PAAM) substrates and utilized AFM to study substrate elasticity and also the adhesion and mechanical properties of endothelial cells in response to changing substrate stiffness. Maximum detachment force and cell stiffness were increased with increasing substrate stiffness. Maximum detachment force values were 0.28 ± 0.14, 0.94 ± 0.27, and 1.99 ± 0.59 nN while Young's moduli of cells were 218.85 ± 38.73, 385.58 ± 131.67, and 933.20 ± 428.92 Pa for soft, medium, and hard substrates, respectively. Human umbilical vein endothelial cells (HUVECs) showed round to more spread shapes on soft to hard substrates, with the most organized and elongated actin structure on the hard hydrogel. Our results confirm the importance of substrate stiffness in regulating cell mechanics and adhesion for a successful cell therapy.

Intermittent hydrostatic pressure enhances growth factor-induced chondroinduction of human adipose-derived mesenchymal stem cells.

Hydrostatic pressure (HP) plays an essential role in regulating function of chondrocytes and chondrogenic differentiation. The objective of this study was to examine effects of intermittent HP on chondrogenic differentiation of human adipose-derived mesenchymal stem cells (hASCs) in the presence or absence of chemical chondrogenic medium. Cells were isolated from abdominal fat tissue and confirmed for expression of ASC surface proteins and differentiation potential. Passage 3 pellets were treated with chemical (growth factor), mechanical (HP of 5 MPa and 0.5 Hz with duration of 4 h/day for 7 consecutive days), and combined chemical-mechanical stimuli. Using real-time polymerase chain reaction, the expression of Sox9, collagen II, and aggrecan as three major chondrogenic markers were quantified among three experimental groups and compared to those of stem cells and human cartilage tissue. In comparison to the chemical and mechanical groups, the chemical-mechanical group showed the highest expression for all three chondrogenic genes close to that of cartilage tissue. Results show the beneficial role of intermittent HP on chondrogenic differentiation of hASCs, and that this loading regime in combination with chondrogenic medium can be used in cartilage tissue engineering.