Biocompatibility and Application of Carbon Fibers in Heart Valve Tissue Engineering

The success of tissue-engineered heart valves rely on a balance between polymer degradation, appropriate cell repopulation, and extracellular matrix (ECM) deposition, in order for the valves to continue their vital function. However, the process of remodeling is highly dynamic and species dependent. The carbon fibers have been well used in the construction industry for their high tensile strength and flexibility and, therefore, might be relevant to support tissue-engineered hearts valve during this transition in the mechanically demanding environment of the circulation. The aim of this study was to assess the suitability of the carbon fibers to be incorporated into tissue-engineered heart valves, with respect to optimizing their cellular interaction and mechanical flexibility during valve opening and closure. The morphology and surface oxidation of the carbon fibers were characterized by scanning electron microscopy (SEM). Their ability to interact with human adipose-derived stem cells (hADSCs) was assessed with respect to cell attachment and phenotypic changes. hADSCs attached and maintained their expression of stem cell markers with negligible differentiation to other lineages. Incorporation of the carbon fibers into a stand-alone tissue-engineered aortic root, comprised of jet-sprayed polycaprolactone aligned carbon fibers, had no negative effects on the opening and closure characteristics of the valve when simulated in a pulsatile bioreactor. In conclusion, the carbon fibers were found to be conducive to hADSC attachment and maintaining their phenotype. The carbon fibers were sufficiently flexible for full motion of valvular opening and closure. This study provides a proof-of-concept for the incorporation of the carbon fibers into tissue-engineered heart valves to continue their vital function during scaffold degradation.

Wnt/β-Catenin Pathway Balances Scaffold Degradation and Bone Formation in Tissue-Engineered Laminae

Tissue engineering provides a promising way for the regeneration of artificial vertebral laminae. Previous studies have confirmed the feasibility of reconstructing vertebral laminae via hydroxyapatite-collagen I scaffolds and mesenchymal stromal cells. However, there were no studies exploring the degradation of hydroxyapatite-collagen I scaffolds and the function of Wnt/β-catenin pathway in the process. In this study, tissue-engineered laminae (TEL) were constructed by nanohydroxyapatite/collagen I scaffolds and umbilical cord Wharton’s Jelly mesenchymal stromal cells (WJ-MSCs). Cell attachment was observed by scanning electron microscopy, and cell viability was confirmed by Live/Dead staining. The rat models were randomly divided into control and β-catenin inhibition groups. Vertebral lamina defect rat models were made on the fifth lumbar vertebrate, and TEL was implanted into the defect site. After 14 weeks, the newborn laminae were harvested for microcomputed tomography, histology, or transcriptional profile analysis. We found that, for the control group, the newborn lamina formation matched with the scaffold degradation and complete newborn laminae formed at the 14th week; for the β-catenin inhibition group, the scaffold degradation rate overrated the lamina formation and no complete artificial laminae were formed at the 14th week. In addition, the osteoclastic genes, such as Cathepsin K or RANKL, in the control groups were significantly lower than the β-catenin inhibition group, and the antiosteoclastic gene, OPG, in the control group was significantly higher than the β-catenin inhibition group. In conclusion, inhibition of Wnt/β-catenin pathway led to speedy scaffold degradation and deferred artificial lamina formation. Wnt/β-catenin pathway played a critical role in maintaining the balance between scaffold degradation and bone formation in the process of vertebral lamina reconstruction.

Engineering Bone with Mesenchymal Stem Cells: Challenges and Obstacles

Repair and reconstruction of large bone defects remain a significant challenge. Cell construct, containing mesenchymal stem cells (MSCs) and scaffold, is a promising strategy for addressing and treating major orthopedic clinical conditions. However, the design of an ideal cell construct for engineering bone faces two critical challenges (i) matching the scaffold degradation rate to that of new bone formation and (ii) preventing the massive cell death post-implantation (caused by disruption of oxygen and nutrient supply). We will hereby primarily focus on the challenge of survival of MSCs post-implantation. Increasing evidence indicates that metabolic regulation plays a critical role in cell fate and functions. In cell metabolism, glucose is considered the major metabolic substrate to produce ATP via glycolysis when the availability of oxygen is limited. In this paper, we delineate the essential roles of glucose on MSC survival. We aim to provide a different perspective which highlights the importance of considering glucose in the development of tissue engineering strategies in order to improve the efficiency of MSC-based cell constructs in the repair of large bone defects.

Application of an ultrasound semi-quantitative assessment in the degradation of silk fibroin scaffolds in vivo

Background Research on the degradation of silk fibroin (SF) scaffolds in vivo lacks uniform and effective standards and experimental evaluation methods. This study aims to evaluate the application of ultrasound in assessing the degradation of SF scaffolds. Methods Two groups of three-dimensional regenerated SF scaffolds (3D RSFs) were implanted subcutaneously into the backs of Sprague-Dawley rats. B-mode ultrasound and hematoxylin and eosin (HE) staining were performed on days 3, 7, 14, 28, 56, 84, 112, 140, and 196. The cross-sectional areas for two groups of 3D RSFs that were obtained using these methods were semi-quantitatively analyzed and compared to evaluate the biodegradation of the implanted RSFs. Results The 3D RSFs in the SF-A group were wholly degraded at the 28th week after implantation. In contrast, the 3D RSFs in the SF-B group were completely degraded at the 16th week. Ultrasonic examination showed that the echoes of 3D RSFs in both groups gradually decreased with the increase of the implantation time. In the early stages of degradation, the echoes of the samples were higher than the echo of the muscle. In the middle of degeneration, the echoes were equal to the echo of the muscle. In the later stage, the echoes of the samples were lower than that of the muscle. The above changes in the SF-B group were earlier than those in the SF-A group. Semi-quantitative analysis of the cross-sectional areas detected using B-mode ultrasound revealed that the degradations of the two 3D RSF groups were significantly different. The degradation rate of the SF-B group was found to be higher than that of the SF-A group. This was consistent with the semi-quantitative detection results for HE staining. Regression analysis showed that the results of the B-mode ultrasound and HE staining were correlated in both groups, indicating that B-mode ultrasound is a reliable method to evaluate the SF scaffold degradation in vivo. Conclusions This study suggests that B-mode ultrasound can clearly display the implanted SF scaffolds non-invasively and monitor the degradation of the different SF scaffolds after implantation in living organisms in real-time.

Fabrication of Dexamethasone-Silver Nanoparticles Entrapped Dendrimer Collagen Matrix Nanoparticles for Dental Applications

A bioengineered tooth is a well-studied subject. However, it is a challenge when seamless integration of various functionalities is required. In the case of teeth, integrating stem cell differentiating agents, regenerating the dentin tissue, and arresting the biofilm formation are three aspects that call for attention. Dental pulp stem cell differentiation to odontoblasts can be facilitated by dexamethasone (Dex). The ideal scaffold for regeneration applications – collagen (Col), has several advantages but also has no inherent rigidity. Silver nanoparticles (AgNps) are found to be an ideal material for preventing biofilm formation, with challenges arising from their stability and cytotoxicity. The route was chosen based on the successful demonstration of various platforms is the PAMAM (Den) dendrimers. The platform encompassing all the ingredients – AgNp@Den@Dex@Col had a max at 622 nm. FTIR studies revealed the presence of all components, and scaffold degradation temperature improved. A linked morphology was observed under TEM, and the zone of inhibition (MIC) was 3.5 mm, indicating good antibacterial activity. There was also a 60% reduction in biofilm formation against Staphylococcus aureus and 40% against Klebsiella pneumoniae by the fabricated nanocomposite AgNp@Den@Dex@Col. In essence, this work reports a platform where three functions of stem cell differentiation, regeneration, and biofilm formation are integrated.

Application of an Ultrasound Semi-Quantitative Assessment in the Degradation of Silk Fibroin Scaffolds In Vivo

Background: Research on the degradation of silk fibroin (SF) scaffolds in vivo lacks uniform and effective standards and experimental evaluation methods. This study aims to evaluate the application of ultrasound in assessing the degradation of SF scaffolds.Methods: Two groups of three-dimensional regenerated SF scaffolds (3D RSFs) were implanted subcutaneously into the backs of Sprague-Dawley rats. B-mode ultrasound and hematoxylin and eosin (HE) staining were performed on days 3, 7, 14, 28, 56, 84, 112, 140, and 196. The cross-sectional areas for two groups of 3D RSFs that were obtained using these methods were semi-quantitatively analyzed and compared to evaluate the biodegradation of the implanted RSFs.Results: Semi-quantitative analysis of the cross-sectional areas detected using B-mode ultrasound revealed that the degradations of the two 3D RSF groups were significantly different. The degradation rate of the SF-B group was found to be higher than that of the SF-A group. This was consistent with the semi-quantitative detection results for HE staining. Regression analysis showed that the results of the B-mode ultrasound and HE staining were correlated in both groups, indicating that B-mode ultrasound is a reliable method to evaluate the SF scaffold degradation in vivo. As the SF scaffold degraded, its echo gradually decreased. In the early stages of degradation, the echo of the SF scaffold was higher than that of the muscle. In the middle stage of degradation, the echo was equal to that of the muscle. In the later stage, the echo was lower than that of the muscle.Conclusions: This study suggests that B-mode ultrasound can clearly display the implanted SF scaffolds non-invasively and monitor the degradation of the different SF scaffolds after implantation in living organisms in real-time.

Impact of Digestive Inflammatory Environment and Genipin Crosslinking on Immunomodulatory Capacity of Injectable Musculoskeletal Tissue Scaffold

The paracrine and autocrine processes of the host response play an integral role in the success of scaffold-based tissue regeneration. Recently, the immunomodulatory scaffolds have received huge attention for modulating inflammation around the host tissue through releasing anti-inflammatory cytokine. However, controlling the inflammation and providing a sustained release of anti-inflammatory cytokine from the scaffold in the digestive inflammatory environment are predicated upon a comprehensive understanding of three fundamental questions. (1) How does the release rate of cytokine from the scaffold change in the digestive inflammatory environment? (2) Can we prevent the premature scaffold degradation and burst release of the loaded cytokine in the digestive inflammatory environment? (3) How does the scaffold degradation prevention technique affect the immunomodulatory capacity of the scaffold? This study investigated the impacts of the digestive inflammatory environment on scaffold degradation and how pre-mature degradation can be prevented using genipin crosslinking and how genipin crosslinking affects the interleukin-4 (IL-4) release from the scaffold and differentiation of naïve macrophages (M0). Our results demonstrated that the digestive inflammatory environment (DIE) attenuates protein retention within the scaffold. Over 14 days, the encapsulated protein released 46% more in DIE than in phosphate buffer saline (PBS), which was improved through genipin crosslinking. We have identified the 0.5 (w/v) genipin concentration as an optimal concentration for improved IL-4 released from the scaffold, cell viability, mechanical strength, and scaffold porosity, and immunomodulation studies. The IL-4 released from the injectable scaffold could differentiate naïve macrophages to an anti-inflammatory (M2) lineage; however, upon genipin crosslinking, the immunomodulatory capacity of the scaffold diminished significantly, and pro-inflammatory markers were expressed dominantly.

Manipulating Air-Gap Electrospinning to Create Aligned Polymer Nanofiber-Wrapped Glass Microfibers for Cortical Bone Tissue Engineering

Osteons are the repeating unit throughout cortical bone, consisting of canals filled with blood and nerve vessels surrounded by concentric lamella of hydroxyapatite-containing collagen fibers, providing mechanical strength. Creating a biodegradable scaffold that mimics the osteon structure is crucial for optimizing cellular infiltration and ultimately the replacement of the scaffold with native cortical bone. In this study, a modified air-gap electrospinning setup was exploited to continuously wrap highly aligned polycaprolactone polymer nanofibers around individual 1393 bioactive glass microfibers, resulting in a synthetic structure similar to osteons. By varying the parameters of the device, scaffolds with polymer fibers wrapped at angles between 5–20° to the glass fiber were chosen. The scaffold indicated increased cell migration by demonstrating unidirectional cell orientation along the fibers, similar to recent work regarding aligned nerve and muscle regeneration. The wrapping decreased the porosity from 90% to 80%, which was sufficient for glass conversion through ion exchange validated by inductively coupled plasma. Scaffold degradation was not cytotoxic. Encapsulating the glass with polymer nanofibers caused viscoelastic deformation during three-point bending, preventing typical brittle glass fracture, while maintaining cell migration. This scaffold design structurally mimics the osteon, with the intent to replace its material compositions for better regeneration.