Surface-Conjugated Galactose on Electrospun Polycaprolactone Nanofibers: An Innovative Scaffold for Uterine Tissue Engineering.

The uterus, a vital organ in the female reproductive system, nurtures and supports developing embryos until maturity. This study focuses on addressing uterine related problems by creating a nanofibrous scaffold to regenerate uterine myometrial tissue, closely resembling the native extracellular matrix (ECM) for enhanced efficacy. To achieve this, we utilized polycaprolactone (PCL) as a biomaterial and employed an electrospinning technique to generate PCL nanofibers in both random and aligned orientations. Due to the inherent hydrophobic nature of PCL nanofibers, a two-step wet chemistry surface modification technique is used, involving the conjugation of galactose onto them. Galactose, a lectin-binding sugar, was chosen to enhance the scaffold's hydrophilicity, thereby improving cell adhesion and fostering l-selectin-based interactions between the scaffold and uterine cells. These interactions, in turn, activated uterine fibroblasts, leading to ECM remodeling. The optimized electrospinning process successfully generated random and aligned nanofibers. Subsequent surface modification was carried out, and the modified scaffold was subjected to various physicochemical characterization, such as the ninhydrin assay, enzyme-linked lectin assay techniques that revealed successful galactose conjugation, and mechanical characterization to assess any changes in material bulk properties resulting from the modification. The tensile strength of random galactose-modified PCL fibers reached 0.041 ± 0.01 MPa, outperforming random unmodified PCL fibers (0.026 ± 0.01 MPa), aligned unmodified PCL fibers (0.011 ± 0.001 MPa), and aligned modified PCL fibers (0.016 ± 0.002 MPa). Cytocompatibility studies with human uterine fibroblast cells showed enhanced viability and proliferation on the modified scaffolds. Initial pilot studies were attempted in the current study involving subcutaneous implantation in the dorsal area of Wistar rats to assess biocompatibility and tissue response before proceeding to intrauterine implantation indicated that the modification did not induce adverse inflammation in vivo. In conclusion, our study introduces a surface-modified PCL nanofibrous material for myometrial tissue engineering, offering promise in addressing myometrial damage and advancing uterine health and reproductive well-being.

Dual Crosslinked Antioxidant Mixture of Poly(vinyl alcohol) and Cerium Oxide Nanoparticles as a Bioink for 3D Bioprinting.

Three-dimensional (3D) bioprinting has made it possible to fabricate structures with intricate morphologies and architectures, which is considered difficult to do when using other conventional techniques like electrospinning. Although the 3D printing of thermoplastics has seen a huge boom in the past few years, it has been challenging to translate this technology to cell-based printing. A major limitation in bioprinting is the lack of inks that allow for the printing of 3D structures that meet the biological requirements of a specific organ or tissue. A bioink is a viscous polymer solution that cells are incorporated into before printing. Therefore, a bioink must have specific characteristics to ensure both good printability and biocompatibility. Despite the progress that has been made in bioprinting, achieving a balance between these two properties has been difficult. In this work, we developed a multimodal bioink that serves as both a cell carrier and a free radical scavenger for treating peripheral nerve injury. This bioink comprises poly(vinyl alcohol) (PVA) and cerium oxide nanoparticles (also called nanoceria (NC)) and was developed with a dual crosslinking method that utilizes citric acid and sodium hydroxide. By employing this dual crosslinking method, good printability of the bioink and shape fidelity of the bioprinted structure were achieved. Additionally, a cell viability study demonstrated that the cells remained compatible and viable even after they underwent the printing process. The combination of this PVA/NC bioink and the dual crosslinking method proved to be effective in enhancing printability and cell biocompatibility for extrusion-based bioprinting applications.

Polycaprolactone/graphene oxide/acellular matrix nanofibrous scaffolds with antioxidant and promyelinating features for the treatment of peripheral demyelinating diseases.

Peripheral demyelinating diseases entail damage to axons and Schwann cells in the peripheral nervous system. Because of poor prognosis and lack of a cure, this group of diseases has a global impact. The primary underlying cause of these diseases involves the inability of Schwann cells to remyelinate the damaged insulating myelin around axons, resulting in neuronal death over time. In the past decade, extensive research has been directed in the direction of Schwann cells focusing on their physiological and neuroprotective effects on the neurons in the peripheral nervous system. One cause of dysregulation in the remyelinating function of Schwann cells has been associated with oxidative stress. Tissue-engineered biodegradable scaffolds that can stimulate remyelination response in Schwann cells have been proposed as a potential treatment strategy for peripheral demyelinating diseases. However, strategies developed to date primarily focussed on either remyelination or oxidative stress in isolation. Here, we have developed a multifunctional nanofibrous scaffold with material and biochemical cues to tackle both remyelination and oxidative stress in one matrix. We developed a nanofibrous scaffold using polycaprolactone (PCL) as a foundation loaded with antioxidant graphene oxide (GO) and coated this bioscaffold with Schwann cell acellular matrix. In vitro studies revealed both antioxidant and remyelination properties of the developed bioscaffold. Based on the results, the developed multifunctional bioscaffold approach can be a promising biomaterial approach for treating demyelinating diseases.

Current status and challenges in uterine myometrial tissue engineering.

The uterus undergoes significant modifications throughout pregnancy to support embryo development and fetal growth. However, conditions like fibroids, adenomyosis, cysts, and C-section scarring can cause myometrial damage. The importance of the uterus and the challenges associated with myometrial damage, and the need for alternative approaches are discussed in this review. The review also explores the recent studies in tissue engineering, which involve principles of combining cells, scaffolds, and signaling molecules to create functional uterine tissues. It focuses on two key approaches in uterine tissue engineering: scaffold technique using decellularized, natural, and synthetic polymer and 3D bioprinting. These techniques create supportive structures for cell growth and tissue formation. Current treatment options for myometrial damage have limitations, leading to the exploration of regenerative medicine and integrative therapies. The review emphasizes the potential benefits of tissue engineering, including more effective and less invasive treatment options for myometrial damage. The challenges of developing biocompatible materials and optimizing cell growth and differentiation are discussed. In conclusion, uterine tissue engineering holds promise for myometrial regeneration and the treatment of related conditions. This review highlights the scientific advancements in the field and underscores the potential of tissue engineering as a viable approach. By addressing the limitations of current treatments, tissue engineering offers new possibilities for improving reproductive health and restoring uterine functionality. Future research shall focus on overcoming challenges and refining tissue engineering strategies to advance the field and provide effective solutions for myometrial damage and associated disorders.

Schwann cell-matrix coated PCL-MWCNT multifunctional nanofibrous scaffolds for neural regeneration.

Nerve tissue engineering aims to create scaffolds that promote nerve regeneration in the damaged peripheral nervous system. However, there remain some challenges in the construction of scaffolds in terms of mechanical properties and cellular behaviour. The present work aims to develop multifunctional implantable nanofibrous scaffolds for nerve regeneration. Using electrospinning, nanofibrous neat polycaprolactone (PCL) and PCL/multiwalled carbon nanotubes (PCL-MWCNT) composite scaffolds were prepared in random and aligned morphology. Schwann cells and their secreted biochemical factors are responsible for neuronal survival in the peripheral nervous system. Therefore, the acellular matrix of Schwann cells was spin-coated on the PCL-MWCNT scaffolds to aid nerve regeneration. Physicochemical and mechanical properties, and the in vitro cellular response of the developed nanofibrous were investigated. We observed no significant change in fibre diameter between neat PCL and PCL-MWCNT scaffolds regardless of the morphology. However, the inclusion of MWCNT reduced the mechanical strength of nanocomposite scaffolds compared to neat PCL. In vitro study revealed biocompatibility of the developed scaffolds both with and without an acellular matrix. Gene expression study revealed a significant increase in peripheral myelin protein (PMP22) expression on acellular matrix-coated PCL-MWCNT scaffolds compared to neat PCL counterparts. Overall, the results suggested Schwann cell matrix-coated PCL-MWCNT nanofibers as a promising conduit for peripheral nerve regeneration.

Silica-Based Bioactive Glasses and Their Applications in Hard Tissue Regeneration: A Review.

Regenerative medicine is a field that aims to influence and improvise the processes of tissue repair and restoration and to assist the body to heal and recover. In the field of hard tissue regeneration, bio-inert materials are being predominantly used, and there is a necessity to use bioactive materials that can help in better tissue-implant interactions and facilitate the healing and regeneration process. One such bioactive material that is being focused upon and studied extensively in the past few decades is bioactive glass (BG). The original bioactive glass (45S5) is composed of silicon dioxide, sodium dioxide, calcium oxide, and phosphorus pentoxide and is mainly referred to by its commercial name Bioglass. BG is mainly used for bone tissue regeneration due to its osteoconductivity and osteostimulation properties. The bioactivity of BG, however, is highly dependent on the compositional ratio of certain glass-forming system content. The manipulation of content ratio and the element compositional flexibility of BG-forming network developed other types of bioactive glasses with controllable chemical durability and chemical affinity with bone and bioactivity. This review article mainly discusses the basic information about silica-based bioactive glasses, including their composition, processing, and properties, as well as their medical applications such as in bone regeneration, as bone grafts, and as dental implant coatings.

Antioxidant for Neurological Diseases and Neurotrauma and Bioengineering Approaches.

Antioxidants are a class of molecules with an innate affinity to neutralize reactive oxygen species (ROS), which are known to cause oxidative stress. Oxidative stress has been associated with a wide range of diseases mediated by physiological damage to the cells. ROS play both beneficial and detrimental roles in human physiology depending on their overall concentration. ROS are an inevitable byproduct of the normal functioning of cells, which are produced as a result of the mitochondrial respiration process. Since the establishment of the detrimental effect of oxidative stress in neurological disorders and neurotrauma, there has been growing interest in exploring antioxidants to rescue remaining or surviving cells and reverse the neurological damage. In this review, we present the survey of different antioxidants studied in neurological applications including neurotrauma. We also delve into bioengineering approaches developed to deliver antioxidants to improve their cellular uptake in neurological applications.

Peptide nanostructures on nanofibers for peripheral nerve regeneration.

Self-assembled peptide nanofibrous scaffolds with designer sequences, similar to neurite growth promoting molecules enhance the differentiation of neural stem cells. However, self-assembled peptide nanofibrous scaffolds lack the required mechanical strength to suffice to bridge long critical-sized peripheral nerve defects. Hence, there is a demand for a potential neural substrate, which could be biomimetic coupled with bioactive nanostructures to regrow the denuded axons towards the distal end. In the present study, we developed designer self-assembling peptide-based aligned poly(lactic-co-glycolic acid) (PLGA) nanofibrous scaffolds by simple surface coating of peptides or coelectrospinning. Retention of secondary structures of peptides in peptide-coated and cospun fibers was confirmed by circular dichroism spectroscopy. The rod-like peptide nanostructures enhance the typical bipolar morphology of Schwann cells. Although the peptide-coated PLGA scaffolds exhibited significant increase in Schwann cell proliferation than pristine PLGA and PLGA-peptide cospun scaffolds (p < .05), peptide cospun scaffolds demonstrated better cellular infiltration and significantly higher gene expression of neural cell adhesion molecule, glial fibrillary acidic protein, and peripheral myelin protein22 compared to the pristine PLGA and PLGA-peptide-coated scaffolds. Our results demonstrate the positive effects of aligned peptide coelectrospun scaffolds with biomimetic cell recognition motifs towards functional proliferation of Schwann cells. These scaffolds could subsequently repair peripheral nerve defects by augmenting axonal regeneration and functional nerve recovery.

Melanin incorporated electroactive and antioxidant silk fibroin nanofibrous scaffolds for nerve tissue engineering.

Nerve restoration and repair in the central nervous system is complicated and requires several factors to be considered while designing the scaffolds like being bioactive as well as having neuroinductive, neuroconductive and antioxidant properties. Aligned electrospun nanofibers provide necessary guidance and topographical cues required for directing the axonal and neurite outgrowth during regeneration. Conduction of nerve impulses is a mandatory feature of a typical nerve. The neuro-conductive property can be imparted by blending the biodegradable, bioactive polymers with conductive polymers. This will provide additional features, i.e., electrical cues to the already existing topographical and bioactive cues in order to make it a more multifaceted neuroregenerative approach. Hence in the present study, we used a combination of silk fibroin and melanin for the fabrication of random and aligned electrospun nanofibrous composite scaffolds. We performed the physico-chemical characterization and also assessed their antioxidant properties. We also evaluated their neurogenic potential using human neuroblastoma cells (SH-SY5Y) for their cellular viability, proliferation, adhesion and differentiation levels. Designed nanofibrous scaffolds had adequate physical properties suitable as neural substrates to promote neuronal growth and regeneration. They stimulated the neuroblastoma cell attachment and viability indicating their biocompatible nature. Silk/melanin composite scaffolds have specifically exhibited high antioxidant nature proven by the radical scavenging activity. Additionally, the melanin incorporated aligned silk fibroin scaffolds promoted the cell differentiation into neurons and orientation along their axis. Our results confirmed the potential of melanin incorporated aligned silk fibroin scaffolds as the promising candidates for effective nerve regeneration and recovery.

Self-assembling peptide nanostructures on aligned poly(lactide-co-glycolide) nanofibers for the functional regeneration of sciatic nerve.

AIM: Regeneration of functional peripheral nerve tissue at critical-sized defect requires extracellular matrix analogs impregnated with appropriate biosignals to regulate the cell fate process and subsequent tissue progression. The aim of the study was to develop electrospun aligned nanofibers as architectural analogs integrated with RADA16-I-BMHP1 as biofunctional peptides. MATERIALS & METHODS: Aligned poly(lactide-co-glycolide) (PLGA)-RADA16-I-BMHP1 nanofibers were fabricated and characterized for their in vitro potential using rat Schwann cell line and in vivo potential using a 10 mm sciatic nerve transection rat model. RESULTS: PLGA-peptide scaffolds significantly promoted higher expression of genotypic markers and bipolar extension of Schwann cells. Further, PLGA-peptide treated animals promoted the native collagen organization, remyelination and showed significantly higher recovery of sensorimotor and motor function than PLGA-treated groups (p < 0.05). CONCLUSION: Our results demonstrate that self-assembling peptide nanostructures on aligned PLGA nanofibers provided better cell-matrix communication with significant functional restoration of the sciatic nerve.

PLGA nanofibers blended with designer self-assembling peptides for peripheral neural regeneration.

Electrospun nanofibers are attractive candidates for neural regeneration due to similarity to the extracellular matrix. Several synthetic polymers have been used but they lack in providing the essential biorecognition motifs on their surfaces. Self-assembling peptide nanofiber scaffolds (SAPNFs) like RADA16 and recently, designer SAPs with functional motifs RADA16-I-BMHP1 areexamples, which showed successful spinal cord regeneration. But these peptide nanofiber scaffolds have poor mechanical properties and faster degradation rates that limit their use for larger nerve defects. Hence, we have developed a novel hybrid nanofiber scaffold of polymer poly(L-lactide-co-glycolide) (PLGA) and RADA16-I-BMHP1. The scaffolds were characterized for the presence of peptides both qualitatively and quantitatively using several techniques like SEM, EDX, FTIR, CHN analysis, Circular Dichroism analysis, Confocal and thermal analysis. Peptide self-assembly was retained post-electrospinning and formed rod-like nanostructures on PLGA nanofibers. In vitro cell compatibility was studied using rat Schwann cells and their adhesion, proliferation and gene expression levels on the designed scaffolds were evaluated. Our results have revealed the significant effects of the peptide blended scaffolds on promoting Schwann cell adhesion, extension and phenotypic expression. Neural development markers (SEM3F, NRP2 & PLX1) gene expression levels were significantly upregulated in peptide blended scaffolds compared to the PLGA scaffolds. Thus the hybrid blended novel designer scaffolds seem to be promising candidates for successful and functional regeneration of the peripheral nerve.

Self-assembling peptide nanofibrous scaffolds for tissue engineering: novel approaches and strategies for effective functional regeneration.

Tissue engineering requires an ideal scaffold that will aid in the regeneration of the damaged tissues both structurally and functionally. Conventionally, polymeric nanofibrous scaffolds have been extensively used due to their structural similarity to the native extracellular matrix. Thus far, top-down approaches like electrospinning and phase separation have been predominantly used for the nanofiber fabrication. Recently, self-assembling peptide nanofibers (SAPNF) have been identified as promising scaffolds for tissue engineering applications. Molecular self-assembly of peptides, which is a bottom-up approach has laid foundations for the development of such novel scaffolds. Designer self-assembling peptides provide functional support as well as bio-recognition due to the presence of bioactive motifs embedded in them. However, there are certain limitations to both electrospun and SAPNF scaffolds in terms of synthesis, cues presented to the biological system and applications. Design of composite, hybrid scaffolds by super-positioning possible cues may result in effective functional tissue regeneration at multiple levels.