Continuous Production of Highly Tuned Silk/Calcium-Based Composites: Exploring New Pathways for Skin Regeneration.

Calcium plays an important role in barrier function repair and skin homeostasis. In particular, calcium phosphates (CaPs) are well established materials for biomedical engineering due to their biocompatibility. To generate biomaterials with a more complete set of biological properties, previously discarded silk sericin (SS) has been recovered and used as a template to grow CaPs. Crucial characteristics for skin applications, such as antibacterial activity, can be further enhanced by doping CaPs with cerium (Ce) ions. The effectiveness of cell attachment and growth on the materials highly depends on their morphology, particle size distribution, and chemical composition. These characteristics can be tailored through the application of oscillatory flow technology, which provides precise mixing control of the reaction medium. Thus, in the present work, CaP/SS and CaP/SS/Ce particles were fabricated for the first time using a modular oscillatory flow plate reactor (MOFPR) in a continuous mode. Furthermore, the biological behavior of both these composites and of previously produced pure CaPs was assessed using human dermal fibroblasts (HDFs). It was demonstrated that both CaP based with plate-shaped nanoparticles and CaP-SS-based composites significantly improved cell viability and proliferation over time. The results obtained represent a first step towards the reinvention of CaPs for skin engineering.

Chitosan and Hydroxyapatite Based Biomaterials to Circumvent Periprosthetic Joint Infections.

Every year, worldwide, millions of people suffering from joint pain undergo joint replacement. For most patients, joint arthroplasty reduces pain and improve function, though a small fraction will experience implant failure. One of the main reasons includes prosthetic joint infection (PJI), involving the prosthesis and adjacent tissues. Few microorganisms (MO) are required to inoculate the implant, resulting in the formation of a biofilm on its surface. Standard treatment includes not only removal of the infected prosthesis but also the elimination of necrotic bone fragments, local and/or systemic administration of antibiotics, and revision arthroplasty with a new prosthesis, immediately after the infection is cleared. Therefore, an alternative to the conventional therapeutics would be the incorporation of natural antimicrobial compounds into the prosthesis. Chitosan (Ch) is a potential valuable biomaterial presenting properties such as biocompatibility, biodegradability, low immunogenicity, wound healing ability, antimicrobial activity, and anti-inflammatory potential. Regarding its antimicrobial activity, Gram-negative and Gram-positive bacteria, as well as fungi are highly susceptible to chitosan. Calcium phosphate (CaP)-based materials are commonly utilized in orthopedic and dentistry for their excellent biocompatibility and bioactivity, particularly in the establishment of cohesive bone bonding that yields effective and rapid osteointegration. At present, the majority of CaP-based materials are synthetic, which conducts to the depletion of the natural resources of phosphorous in the future due to the extensive use of phosphate. CaP in the form of hydroxyapatite (HAp) may be extracted from natural sources as fish bones or scales, which are by-products of the fish food industry. Thus, this review aims to enlighten the fundamental characteristics of Ch and HAp biomaterials which makes them attractive to PJI prevention and bone regeneration, summarizing relevant studies with these biomaterials to the field.

A biocompatible and injectable hydrogel to boost the efficacy of stem cells in neurodegenerative diseases treatment.

AIMS: Stem cell therapies emerged as treatment modalities with potential to cure neurodegenerative diseases (NDs). However, despite high expectations, their clinical use is still limited. Critical issues in treatment outcomes may be related to stem cells formulation and administration route. We develop a hydrogel as a cell carrier, consisting of compounds (phospholipids and hyaluronic acid-HA) naturally present in the central nervous system (CNS). The HA-based hydrogel physically crosslinked with liposomes is designed for direct injection into the CNS to significantly increase the bone marrow mesenchymal stem cells (BMSCs) bioavailability. MATERIALS AND METHODS: Hydrogel compatibility is confirmed in vitro with BMSCs and in vivo through its intracerebroventricular injection in rats. To assess its efficacy, the main cause of chronic neurologic disability in young adults is selected, namely multiple sclerosis (MS). The efficacy of the developed formulation containing a lower number of cells than previously reported is demonstrated using an experimental autoimmune encephalomyelitis (EAE) rat model. KEY FINDINGS: The distribution of the engineered hydrogel into corpus callosum can be ideal for NDs treatment, since damage of this white matter structure is responsible for important neuronal deficits. Moreover, the BMSCs-laden hydrogel significantly decreases disease severity and maximum clinical score and eliminated the relapse. SIGNIFICANCE: The engineering of advanced therapies using this natural carrier can result in efficacious treatments for MS and related debilitating conditions.

In Situ Forming Silk Sericin-Based Hydrogel: A Novel Wound Healing Biomaterial.

In situ cross-linked hydrogels have the advantage of effectively fulfilling the wound in its shape and depth. Amongst the new generation of natural-based biopolymers being proposed for wound care and skin regeneration, silk sericin is particularly interesting due to its exceptional properties such as biocompatibility, biodegradability, and antioxidant behavior, among others. In this study, a new enzyme-mediated cross-linked hydrogel composed of silk sericin is proposed for the first time. The developed hydrogel cross-linking strategy was performed via horseradish peroxidase, under physiological conditions, and presented gelling kinetics under 3 min, as demonstrated by its rheological behavior. The hydrogels presented a high degree of transparency, mainly due to their amorphous conformation. Degradation studies revealed that the hydrogels were stable in phosphate buffer solution (PBS) (pH 7.4) for 17 days, while in the presence of protease XIV (3.5 U/mg) and under acute and chronic physiological pH values, the stability decreased to 7 and 4 days, respectively. During protease degradation, the present sericin hydrogels demonstrated antioxidant activity. In vitro studies using an L929 fibroblast cell line demonstrated that these hydrogels were noncytotoxic, promoting cell adhesion and massive cell colonization after 7 days of culture, demonstrating that cells maintained their viability and proliferation. In addition, the application of sericin-based hydrogel in an in vivo diabetic wound model validated the feasibility of the in situ methodology and demonstrated a local anti-inflammatory effect, promoting the healing process. This study presents a simple, fast, and practical in situ approach to produce a sericin-based hydrogel able to be applied in low exudative chronic wounds. Moreover, the study herein reported fosters the valorization of a textile industrial by-product by its integration in the biomedical field.

Roles of the HOX Proteins in Cancer Invasion and Metastasis.

Invasion and metastasis correspond to the foremost cause of cancer-related death, and the molecular networks behind these two processes are extremely complex and dependent on the intra- and extracellular conditions along with the prime of the premetastatic niche. Currently, several studies suggest an association between the levels of HOX genes expression and cancer cell invasion and metastasis, which favour the formation of novel tumour masses. The deregulation of HOX genes by HMGA2/TET1 signalling and the regulatory effect of noncoding RNAs generated by the HOX loci can also promote invasion and metastasis, interfering with the expression of HOX genes or other genes relevant to these processes. In this review, we present five molecular mechanisms of HOX deregulation by which the HOX clusters products may affect invasion and metastatic processes in solid tumours.

In situ Enabling Approaches for Tissue Regeneration: Current Challenges and New Developments.

In situ tissue regeneration can be defined as the implantation of tissue-specific biomaterials (by itself or in combination with cells and/or biomolecules) at the tissue defect, taking advantage of the surrounding microenvironment as a natural bioreactor. Up to now, the structures used were based on particles or gels. However, with the technological progress, the materials' manipulation and processing has become possible, mimicking the damaged tissue directly at the defect site. This paper presents a comprehensive review of current and advanced in situ strategies for tissue regeneration. Recent advances to put in practice the in situ regeneration concept have been mainly focused on bioinks and bioprinting techniques rather than the combination of different technologies to make the real in situ regeneration. The limitation of conventional approaches (e.g., stem cell recruitment) and their poor ability to mimic native tissue are discussed. Moreover, the way of advanced strategies such as 3D/4D bioprinting and hybrid approaches may contribute to overcome the limitations of conventional strategies are highlighted. Finally, the future trends and main research challenges of in situ enabling approaches are discussed considering in vitro and in vivo evidence.

Scaffolds based bone tissue engineering: the role of chitosan.

As life expectancy increases, malfunction or loss of tissue caused by injury or disease leads to reduced quality of life in many patients at significant socioeconomic cost. Even though major progress has been made in the field of bone tissue engineering, present therapies, such as bone grafts, still have limitations. Current research on biodegradable polymers is emerging, combining these structures with osteogenic cells, as an alternative to autologous bone grafts. Different types of biodegradable materials have been proposed for the preparation of three-dimensional porous scaffolds for bone tissue engineering. Among them, natural polymers are one of the most attractive options, mainly due to their similarities with extracellular matrix, chemical versatility, good biological performance, and inherent cellular interactions. In this review, special attention is given to chitosan as a biomaterial for bone tissue engineering applications. An extensive literature survey was performed on the preparation of chitosan scaffolds and their in vitro biological performance as well as their potential to facilitate in vivo bone regeneration. The present review also aims to offer the reader a general overview of all components needed to engineer new bone tissue. It gives a brief background on bone biology, followed by an explanation of all components in bone tissue engineering, as well as describing different tissue engineering strategies. Moreover, also discussed are the typical models used to evaluate in vitro functionality of a tissue-engineered construct and in vivo models to assess the potential to regenerate bone tissue are discussed.

Adhesion, proliferation, and osteogenic differentiation of a mouse mesenchymal stem cell line (BMC9) seeded on novel melt-based chitosan/polyester 3D porous scaffolds.

The aim of the present work was to study the biological behavior of a mouse mesenchymal stem cell line when seeded and cultured under osteogenic conditions onto novel processed melt-based chitosan scaffolds. Scaffolds were produced by compression molding, followed by salt leaching. Scanning electron microscopy (SEM) observations and microCT analysis showed the pore sizes ranging between 250 and 500 microm and the interconnectivity of the porous structure. The chitosan-poly(butylenes succinate) scaffolds presented high mechanical properties, similar to the ones of trabecular bone (E1% approximately 75 MPa). Cytotoxicity assays were carried out using standard tests (accordingly to ISO/EN 10993 part 5 guidelines), namely, MTS test with a 24 h extraction period, revealing that L929 cells had similar metabolic activities to that obtained for the negative control. Cell culture studies were conducted using a mouse mesenchymal stem cell line (BMC9). Cells were seeded onto the scaffold and allowed to proliferate for 3 weeks under osteogenic conditions. SEM observations demonstrated that cells were able to proliferate and massively colonize the scaffolds structure. The cell viability assay MTS demonstrated that BMC9 cells were viable after 3 weeks of culture. The cells clearly evidenced a positive differentiation toward the osteogenic lineage, as confirmed by the high ALP activity levels. Moreover, energy dispersive spectroscopy (EDS) analysis revealed the presence of Ca and P in the elaborated extracellular matrix (ECM). These combined results indicate that the novel melt-based chitosan/polyester scaffolds support the adhesion, proliferation, and osteogenic differentiation of the mouse MSCs and shows adequate physicochemical and biological properties for being used as scaffolds in bone tissue engineering-related strategies.