Nanofiber Scaffolds Research Articles

Adipose Derived-Stem Cells and their Cell-Free Supernatant Combined with a Nanofiber Scaffold for Skin Rejuvenation: The Avancell Stem Cell SecretomeTM - A Technical Note

The skin is the largest body organ and acts as the first defense line against various external agents and stimuli such as toxins, ultraviolet radiation or environmental agents

Multifunctional silk fibroin and cellulose acetate composite nanofibers incorporated with palladium and platinum nanoparticles for enhanced wound healing: Comprehensive characterization and in vivo assessment

Wound healing is a complex and dynamic process that relies on various factors for optimal tissue regeneration. Current wound management methods often lack the efficiency needed to address these challenges. In this study, we prepared and conducted a comprehensive analysis of silk fibroin and cellulose acetate composite nanofibers incorporated with palladium (Pd) and platinum (Pt) nanoparticles, emphasizing their potential as advanced materials for wound healing. Electron microscopy reveals nanofibrous scaffolds with interconnected fibers, an average diameter of 168.95 ± 135.79 nm, and Pd and Pt nanoparticles of 10.64 nm and 9.21 nm, with thicknesses ranging from 1.48 nm to 2.13 nm. The analysis of swelling, degradation, porosity, and mechanical strength highlights their capacity to absorb exudates from wound surfaces, promote cell adhesion, and support tissue regeneration. In vitro evaluation demonstrates significant antibacterial activity against Escherichia coli and Staphylococcus aureus, with inhibition zone values of 16 mm and 18 mm, respectively. Cell migration, proliferation, and biocompatibility are assessed using the L929 fibroblast cell line with the ethidium bromide (EtBr) dye. In vivo studies on Sprague-Dawley rats indicate SF/CA/Pd-Pt composite nanofibers have exceptional wound healing efficiency, reaching 99.31% compared to the SF/CA matrix. High hemocompatibility, with erythrocyte percentages ranging from 1.6% to 4.1%, further supports these nanofibers wound healing properties. Histopathological analysis, employing hematoxylin and eosin staining, provides valuable insights into tissue regeneration, assessing reepithelialization, granulation, angiogenesis, and reduced inflammation. These findings emphasize the potential of Pd and Pt nanoparticles incorporated into SF/CA composite nanofiber scaffolds as advanced biomedical materials, particularly for wound healing applications.

Catalysis-based approaches with biopolymers and violet LED to improve in-office dental bleaching.

This in vitro experimental investigation aimed to evaluate the impact of the combined application of a nanofiber scaffold (NS), a polymeric catalyst primer (PCP) containing 10mg/mL of heme peroxidase enzyme, and violet LED (LEDv) on the esthetic efficacy (EE), trans-amelodentinal cytotoxicity (TC), and procedural duration of conventional in-office bleaching therapy. To achieve this, 96 standardized enamel/dentin discs were individually placed in artificial pulp chambers. A 35% hydrogen peroxide (H2O2) bleaching gel was administered for 45, 30, or 15min to the enamel, either previously coated with NS + PCP or left uncoated, followed by irradiation with LEDv for 15min or no irradiation. The established groups were as follows: G1, negative control (no treatment); G2, 35% H2O2/45min; G3, NS + PCP + LEDv; G4, NS + PCP + 35%H2O2/45min + LEDv; G5, NS + PCP + 35%H2O2/30min + LEDv; and G6, NS + PCP + 35%H2O2/15min + LEDv. Extracts (culture medium + gel components diffused through the discs) were collected and applied to odontoblast-like MDPC-23 cells. EE (ΔE00 and ΔWI) and TC were assessed using ANOVA/Tukey analysis (p < 0.05). The EE analysis revealed no statistical differences between G6 and G2 (p > 0.05). Cells in G6 exhibited higher viability and lower oxidative stress compared to other bleached groups (p < 0.05). In conclusion, employing NS + PCP + LEDv to catalyze a 35%H2O2 bleaching gel applied for 15min to the enamel resulted in successful esthetic improvements and reduced the cytotoxicity commonly linked with traditional in-office bleaching procedures.

PLATELET-DERIVED EXTRACELLULAR VESICLES PROMOTE STEM CELL TENOGENIC COMMITMENT IN A BIOENGINEERED TENDON 3D MODEL

Worldwide, tendon disorders are one of the main causes of disability that decrease the quality of life of individuals and represent a substantial economic burden on society. Currently, the main therapies used for tendon injuries are not able to restore tendon functionality, and due to tendons' hypovascular and hypocellular nature, they present a reduced healing capacity, which also limits the success of the available therapies. In order to discover new therapies, extracellular vesicles (EVs), key players in cell-cell communication, have been widely explored for tissue engineering and regenerative medicine applications. Thus, the aim of this study is to assess the role of EVs derived from platelets in stem cell tenogenic commitment using a bioengineered tendon in vitro model for potential use as tendon therapeutic agents. Biomimetic platelet-derived EVs were produced by freeze-thaw cycles of platelets and isolation at different centrifugation speed. To recreate the architecture of tendons, a 3D system consisting of electrospun anisotropic nanofiber scaffolds coated with collagen encapsulating human adipose stem cells (hASCs) and different types of platelet-derived EVs, were produced. Then, the influence of the tendon-mimetic constructs and the distinct EVs populations in the hASCs tenogenic differentiation were assessed over culture time. We observed that the hASCs on the nanofibrous tendon scaffolds, show high cytoskeleton anisotropic organization that is characteristic of tenocytes. Moreover, acting as biological cues, platelet-derived EVs boosted hASCs tenogenic commitment, supported by the increased gene expression of tendon-related markers (SCX and TNMD). Additionally, EVs enhanced the deposition of tendon like extracellular matrix (ECM), as evidenced by the increased gene expression of ECM-related markers such as COL1, COL3, DCN, TNC, and MMP-3, which are fundamental for ECM synthesis and degradation balance. Moreover, EVs induced lower collagen matrix contraction on hASCs, which has been related with lower myofibroblast differentiation. Overall, the results revealed that EVs are capable of modulating stem cells' behavior boosting their tenogenic commitment, through the increased expression of healthy tendon cell markers, potentiating ECM deposition and decreasing cell contractility. Therefore, platelet EVs are a promising biochemical tool, worthy to be further explored, as paracrine signaling that might potentiate tendon repair and regeneration.

Biobran-loaded core/shell nanofibrous scaffold: a promising wound dressing candidate.

This research examined the effectiveness of Biobran as a bioactive substance that could potentially improve wound healing. It also looked at how Biobran affects the properties of a nanofibrous scaffold made through coaxial electrospinning. This is the first study exploring the use of Biobran in this context and its interaction with nanofibrous scaffolds. The scaffolds were composed of poly(ε-caprolactone) (PCL) in the shell and various concentrations of Biobran blended with polyvinyl alcohol (PVA) in the core. The properties of the scaffolds were characterized by SEM, TEM, FTIR, XRD, TGA, DSC, stress-strain test, WCA, release test, MTT cytotoxicity assay, wound scratching assay, and the dye exclusion method using trypan blue. The scaffolds loaded with Biobran exhibited a more compact and smooth morphology compared with the scaffold without Biobran. The physical interaction and crystallinity of the polymers in the scaffolds were also affected by Biobran in a concentration-dependent manner. This positively influenced their tensile strength, elongation at break, thermal stability, and hydrophilicity. The porosity, water uptake capacity, and WVTR of the nanofibrous scaffolds are within the optimal ranges for wound healing. The release rate of Biobran, which revealed a biphasic release pattern, decreased with increasing Biobran concentration, resulting in controlled and sustained delivery of Biobran from the nanofiber scaffolds. The cell viability assays showed a dose-dependent effect of Biobran on WISH cells, which might be attributed to the positive effect of Biobran on the physicochemical properties of the nanofibrous scaffolds. These findings suggest that Biobran-loaded core/shell nanofiber scaffolds have a potential application in wound healing as an ideal multifunctional wound dressing.

Fabrication and biocompatibility evaluation of hydroxyapatite-polycaprolactone-gelatin composite nanofibers as a bone scaffold.

One approach to addressing bone defects involves the field of bone tissue engineering, with scaffolds playing an important role. The properties of the scaffold must be similar to those of natural bone, including pore size, porosity, interconnectivity, mechanical attributes, degradation rate, non-toxicity, non-immunogenicity, and biocompatibility. The primary goals of this study are as follows: first, to evaluate hydroxyapatite (HA)/polycaprolactone (PCL)/gelatin nanofiber scaffolds based on functional groups, fibre diameter, porosity, and degradation rate; second, to investigate the interaction between HA/PCL/gelatin scaffolds and osteoblast cells (specifically, the ATCC 7F2 cell line) using in vitro assays, including cell viability and adhesion levels. The fibre samples were fabricated using an electrospinning technique with a 15 kV voltage, a spinneret-collector distance of 10 cm, and a flow rate of 0.3 mL hour-1. The process was applied to five different HA/PCL/gelatin concentration ratios: 50 : 40 : 10; 50 : 30 : 20; 50 : 25 : 25; 50 : 20 : 30; 50 : 35 : 15 (in %wt). Fourier Transform Infrared (FTIR) spectrum analysis and tests revealed no differences in functional groups across the five compositions. The identified functional groups include PO4 3-, OH-, CO3 2- and C[double bond, length as m-dash]O stretching. Notably, an increase in PCL concentrations resulted in larger fiber diameters, ranging from 369-1403 nm with an average value of 929 ± 175 nm. The highest porosity percentage was (77.27 ± 11.57) %, and a sufficient degradation rate of up to 3.5 months facilitated the proliferation process of osteoblast cells. Tensile strength assessments revealed a significant increase in tensile strength with the addition of PCL, reaching a peak of 1.93 MPa. The MTT assay demonstrated a discernible increase in cell proliferation, as evidenced by increased cell viability percentages on days 1, 3, and 5. Concurrently, the fluorescence microscopy examination indicated an increase in cell numbers, which was especially noticeable on days 1 and 5. The SEM analysis confirmed the biocompatibility of the HA/PCL/gelatin nanofiber scaffold, as osteoblast cells attached and dispersed successfully five days after seeding. Based on these findings, the HA/PCL/gelatin nanofiber scaffold emerges as a very promising candidate for treating bone damage.

Polyvinyl alcohol/Polyvinylpyrrolidone/Chitosan Nanofiber Scaffold with Hydroxyapatite from Sand Lobster Shells (Panulirus homarus) for Bone Tissue Engineering

In this work, nanofiber scaffold membrane polyvinyl alcohol (PVA)/polyvinylpyrrolidone (PVP)/Chitosan (CS)/hydroxyapatite (HAp) from sand lobster (SL; Panulirus homarus) shells have successfully synthesized to mimic the extracellular matrix (ECM) nanoscale in the native bone. HAp was synthesized by co-precipitation method with Ca/P was 1.67, then nanofiber membrane PVA/PVP/CS/HAp was synthesized by electrospinning method. Nanofiber solution was prepared from PVA 10% (w/v) polymer solution that dissolved in the distilled water, then the PVP/CS 15% (w/v) polymer solution was dissolved in acetic acid 1% (v/v) separately. The PVA polymer solution and PVP/CS solution were mixed with a ratio of 8.5: 1.5 (v/v). HAp dispersed into mixture solution with variation concentration 0, 1, 3, and 5 wt%. The composite solution was put into a 10 ml syringe with a hole diameter = 0.5 mm. Electrospinning was carried out at a 10 kV voltage, the flow rate at 0.1 ml/h, and the distance between the collector to the tip was 12 cm. Nanofiber scaffold membrane was characterized using SEM, FTIR, and XRD. The addition of HAp into the fiber showed incorporation into nanofiber with small agglomeration in the concentration of HAp at 1, 3, and 5 wt%. Based on the physicochemical analysis, the nanofiber scaffold PVA/PVP/CS/HAp 5 wt% with a fiber diameter of 0.328 ± 0.049 μm has the most potential to be used for bone tissue engineering.

Synthesis and characterization of new electrospun medical scaffold-based modified cellulose nanofiber and bioactive natural propolis for potential wound dressing applications.

Recently, the design of polymer nanofibers using the electrospinning process has attracted much interest. Particularly the use of natural polymers has promoted many advantages in their biomedical applications. However, the combination of multiple natural polymers remains a great challenge in terms of electrospun production and applied performance. From this perspective, the current investigation highlights the study of the preparation of electrospun nanomaterial scaffolds based on combined natural polymers for improved wound healing performance. First, we have synthesized a crosslinked polymer by reacting microcrystalline cellulose (MC) and chitosan (CS) biopolymer via the intermediate of citric acid as a crosslinking agent. Then a natural propolis biomolecule was incorporated into the polymer network. Different MC/CS blend ratios of 90/10 and 70/30 were then used and various machine parameters were optimized to obtain nanofiber scaffolds with excellent strength and structures. SEM, IR, physicochemical, mechanical, and morpho-logical characterization were then performed. SEM evaluation revealed homogeneous and bead-free nanofibrous structures, with well-defined morphology and a random deposition that could accurately mimic the extracellular matrix of native skin. The calculated average nanofiber diameters for the MC/CS blend ratios at 90/10 and 70/30 were 431.4 and 441.2 nm, respectively. The results showed that when the chitosan amount increased, larger nanofibers with narrow diameter distribution appeared. The prepared nanomaterials had a significant and close water vapor permeability of about 1735.12 and 1698.52 g per m per day for the two blend ratios of 90/10 and 70/30, respectively. The examination of swelling behavior revealed a noteworthy enhancement in hydrophilicity, a necessary attribute for improved healing efficacy. FT-IR analysis confirmed the success and the stability of the chemical crosslinking reaction between the two biopolymers before nanofiber conception. Excellent mechanical properties were acquired, based on the chitosan content. Both developed nanofiber scaffolds exhibited high tensile strength and Young's modulus values. The incorporation of 30% chitosan versus 10% results in an increase in tensile strength of 11% and 14% in Young's modulus. Therefore, we could adjust the different mechanical properties simply by varying the mixing rate of the electrospun polymers. Using epithelial HepG2 cells, viability and kinetic cell adhesion assays were assessed to obtain biological evaluation. No cytotoxicity was observed and good cytocompatibility was confirmed. Functionalized nanofiber biomaterials with different MC/CS ratios substantiated significant bactericidal effectiveness against Gram-positive and Gram-negative bacterial culture strains. The novel functional electrospun wound dressing scaffold demonstrated effective and promising biomedical performance, healing both acute and chronic wounds.

Electrospun nanofibers: building blocks for the repair of bone tissue.

Bone, one of the hardest structures of the body, is the basic constituent of the skeletal system, which gives the shape to the body, provides mechanical support for muscles and soft tissues, and provides movement. Even if there is no damage, bone remodeling is a permanent process to preserve and renew the structural, biochemical, and biomechanical integrity of bone tissue. Apart from the remodeling, bone healing is the highly complicated process of repairing deficiencies of bone tissue by the harmonious operation of osteoblasts, osteocytes, osteoclasts, and bone lining cells. Various materials can be used to both trigger the bone healing process and to provide mechanical support to damaged bone. Nanofiber scaffolds are at the forefront of these types of systems because of their extremely large surface area-to-volume ratio, small pore size, and high porosity. Nanofibers are known to be highly functional systems with the ability to mimic the structure and function of the natural bone matrix, facilitating osteogenesis for cell proliferation and bone regeneration. Electrospinning is an easy and fast method to produce non-woven structures consisting of continuous ultrafine fibers with diameters ranging from micrometers down to nanometers. The simplicity and cost-effectiveness of the electrospinning technique, its ability to use a wide variety of synthetic, natural, and mixed polymers, and the formation of highly porous and continuous fibers are the remarkable features of this method. The importance of nanofiber-based scaffolds in bone tissue regeneration is increasing because of suitable pore size, high porosity, osteoinduction, induction of bone growth with osteoconduction, adaptability to the target area, biodegradation, and appropriate mechanical properties, which are among the main parameters that are important in the design of polymeric bone grafts. The aim of this review is to cast light on the increasing use of nanofiber-based scaffolds in bone tissue regeneration and give an insight about bone regeneration, production techniques of the electrospun nanofibers, and varying formulation parameters in order to reach different drug delivery goals. This review also provides an extensive market research of electrospun nanofibers and an overview on scientific research and patents in the field.

Evaluation of Random and Aligned Polycaprolactone Nanofibrous Electrospun Scaffold for Human Periodontal Ligament Engineering in Biohybrid Titanium Implants.

Background: Stem cells are introduced to regenerate some living tissue to restore function and longevity. The study aims to isolate in vitro human periodontal ligament stem cells (hPDLSCs) and investigate their proliferation rate on plasma-treated aligned and random polycaprolactone (PCL) nanofibrous scaffolds made via an electrospinning technique to attempt periodontal-like tissue in dental implants. Materials and Methods: hPDLSCs were isolated from extracted human premolars and cultured on plasma-treated or untreated PCL-aligned and random scaffolds to enhance adhesion of periodontal ligament (PDL) cells as well as interaction and proliferation. Cell morphology, adhesion, and proliferation rate were evaluated using field emission scanning electron microscopy (FESEM) and the methyl tetrazolium (MTT) assay. The wettability of PCL scaffolds was tested using a goniometer. Results: The hydrophilicity of plasma-treated scaffolds was significantly increased (p ≤ 0.05) in both aligned and random nanofibers compared to the nontreated nanofibrous scaffold. Cells arranged in different directions on the random nanofiber scaffold, while for aligned scaffold nanofibers, the cells were arranged in a pattern that followed the direction of the aligned electrospun nanofibres. The rate of hPDLSC proliferation on an aligned PCL nanofiber scaffold was significantly higher than on a random PCL nanofibrous scaffold with a continuous, well-arranged monolayer of cells, as shown in FESEM. Conclusion: The aligned PCL nanofiber scaffold is superior to random PCL when used as an artificial scaffold for hPDLSC regeneration in PDL tissue engineering applications.

Evaluation of osteoconductive effect of polycaprolactone (PCL) scaffold treated with fibronectin on adipose-derived mesenchymal stem cells (AD-MSCs).

Replacing damaged organs or tissues and repairing damage by tissue engineering are attracting great interest today. A potentially effective method for bone remodeling involves combining nanofiber scaffolds with extracellular matrix (ECM), and growth factors. Today, electrospun PCL-based scaffolds are widely used for tissue engineering applications. In this study, we used an electrospun polycaprolactone (PCL) scaffold coated with fibronectin (Fn), a ubiquitous ECM glycoprotein, to investigate the induction potential of this scaffold in osteogenesis with adipose-derived mesenchymal stem cells (AD-MSCs). Scanning electron microscopy (SEM) analysis showed that fibronectin, by binding to the membrane receptors of mesenchymal stem cells (MSCs), leads to their attachment and proliferation on the PCL scaffold and provides a suitable environment for osteogenesis. In addition, biochemical tests showed that fibronectin leads to increased calcium deposition. The results also showed that alkaline phosphatase activity was significantly higher in the PCL scaffold coated with fibronectin than in the control groups (PCL scaffold group and tissue culture polystyrene (TCPS) group) (P<0.05). Also, the analysis of quantitative reverse transcription PCR (qRT-PCR) data showed that the relative expression of bone marker genes such as osteonectin (ON), osteocalcin (OC), RUNX family transcription factor 2 (RUNX2), and collagen type I alpha 1 (COL1) was much higher in the cells seeded on the PCL/Fn scaffold than in the other groups (P<0.05). The results show that fibronectin has an increasing effect in accelerating bone formation and promising potential for use in bone tissue engineering.

Desferrioxamine-Laden Nanofibrous Scaffolds with Efficient Angiogenesis for Accelerating Diabetic Wound Healing.

Delayed diabetic wound healing is one of the clinical difficulties, the main reason is the limited angiogenesis ability. Deferriamine (DFO) is an iron chelating agent that can induce angiogenesis, but its application is limited due to its short half-life. Increasing the load and slow release performance of desferriamine is beneficial to accelerate diabetic wound healing. In this study, we developed collagen (Col)-graphene oxide (GO) and (1% w/w) DFO-loaded nanofiber electrospinning scaffolds (DCG) using the electrospinning technique. We tested the physicochemical properties, drug release performance, and vascularization biological function of the scaffolds, and finally evaluated the promotion of full-thickness wound healing in the diabetic rat models. The results showed that DCG scaffolds have good mechanical properties and water-holding capacity and can release DFO continuously for 14 days. In vitro, the novel DCG scaffold exhibited good biocompatibility, with the up-regulation at the gene level of VEGF and its regulator HIF-1α, promoters of angiogenesis. This was verified in vivo, as the scaffold enhanced granulation tissue formation and improved neovascularization, thereby accelerating wound healing when applied to full-thickness defects on the back of diabetic rats. The DCG nanofiber scaffold prepared in this study has good biocompatibility and vascularization ability, and improves the microenvironment in vivo, and has a good application prospect in diabetic wound repair.

Carboxymethyl chitosan-based hydrogel-Janus nanofiber scaffolds with unidirectional storage-drainage of biofluid for accelerating full-thickness wound healing

Self-pumping wound scaffolds designed for directional biofluid transport are extensively investigated. They efficiently extract excessive biofluids from wounds, while maintaining an optimally humid wound environment, thus facilitating rapid wound healing. However, the existing designed scaffolds are insufficiently focused on stimulating the hydrophobic layer at the wound site, thereby exacerbating inflammation and impeding the wound healing process. Herein, we engineered and fabricated a hydrophilic-hydrophobic-hydrophilic sandwich-structured hydrogel-Janus nanofiber scaffold (NFS) employing a Layer-by-Layer (LbL) method. This scaffold comprises a hydrophilic carboxymethyl chitosan/silver (CMCS-Ag) hydrogel component in conjunction with a poly(caprolactone)/poly(caprolactone)-poly(citric acid)-co-ε-polylysine (PCL/PCL-PCE) Janus NFS. It is noteworthy that the hydrogel-Janus nanofiber scaffold not only demonstrates outstanding water absorption (202.2 %) and unidirectional biofluid transport capability but also possesses high breathability (308.663 m3/m2 h kPa), appropriate pore size (6.7–7.5 μm), excellent tensile performance (270 ± 10 %), and superior mechanical strength (26.36 ± 1.77 MPa). Moreover, in vitro experimentation has convincingly demonstrated the impeccable biocompatibility of hydrogel-Janus NFS. The inherent dual-antibacterial properties in CMCS-Ag and PCE significantly augment fibroblast proliferation and migration. In vivo studies further underscore its capability to expedite wound healing by absorption and expulsion of wound exudates, thereby fostering collagen deposition and vascularization. As such, this work potentially provides fresh insights into the design and fabrication of multifunctional biomimetic scaffolds, holding immense potential in the medical field for efficient wound healing.

Synthesis of bone biocompatible implants using human adipose-derived mesenchymal stem cells (hADMSCs) and PCL/laminin scaffold substrate.

Bone tissue engineering is considered a new method in the treatment of bone defects and can be an effective alternative to surgery and bone grafting. The use of adipose tissue mesenchymal stem cells (ADMSCs) on synthetic polymer scaffolds is one of the new approaches in bone tissue engineering. In this study, we aimed to investigate the effect of laminin coating on biocompatibility and osteogenic differentiation of ADMSCs seeded on polycaprolactone (PCL) scaffolds. The morphology of the electrospun scaffold was evaluated using a scanning electron microscope (SEM). Cell proliferation and cytotoxicity were determined by MTT assay. The adipogenic and osteogenic differentiation potential of the cells was evaluated. The osteogenic differentiation of ADMSCs cultured on the PCL scaffold coated with laminin was assessed by evaluating the level of alkaline phosphatase (ALP) activity, intracellular calcium content, and expression of bone-specific genes. The results showed that the ADMSCs cultured on PCL/laminin showed enhanced osteogenic differentiation compared to those cultured on non-coated PCL or control medium (P<0.05). It seems that laminin enhances the physicochemical properties and biocompatibility of PCL nanofiber scaffolds; and by modifying the surface of the scaffold, improves the differentiation of ADMSCs into osteogenic cells.

FGF-2 enriched nanofiber scaffold for advancing achilles tendon healing: a comparative experimental investigation.

Achilles tendon rupture is a common and debilitating injury that significantly impacts mobility and quality of life. Effective treatment options that promote faster and more complete healing are needed. Fibroblast growth factor-2 (FGF-2) has shown potential in enhancing tendon repair. This study aims to investigate the efficacy of FGF-2 in promoting tendon healing in a rat model of Achilles tendon rupture, providing insights into its potential as a therapeutic option. Forty-eight rat hind legs with complete Achilles tendon ruptures were divided into four equal groups: the Sham (S) group (tendon repair only), the Polymer (P) group (tendon repair with scaffold wrapping), the Produced FGF-2 (PF) group (scaffold coated with lab-produced FGF-2), and the Commercial FGF-2 (CF) group (scaffold coated with commercially sourced FGF-2). Histological analyses at two and four weeks post-surgery evaluated healing based on nuclear morphology, vascularity, fibril organization, inflammation, and adipogenesis. At the end of the second week, no macroscopic healing was observed in one rat each from the S and P groups. By the end of the fourth week, macroscopic healing was observed in all groups. The S and P groups exhibited similarly severe fibril disorganization, pathological adipogenesis, and sustained inflammation, particularly at the fourth week. In contrast, the CF group demonstrated improved tendon healing with increased vascularity and extracellular matrix, lower inflammatory cell infiltration, and better fibril organization. Pathological adipogenesis was absent in the CF group, especially at the fourth week. The PF group showed comparable improvements at the second week but experienced a relapse by the 4th week, with increased inflammation and adipogenesis. FGF-2 coated scaffolds significantly enhanced tendon healing in a rat Achilles tendon rupture model by improving fibril organization, increasing vascularity, and reducing inflammation and pathological adipogenesis. These findings suggest that FGF-2 could be a promising therapeutic option for accelerating tendon repair. Future perspectives on tendon repair will focus on enhancing FGF-2 delivery using innovative scaffolds, paving the way for more effective therapies and improved patient outcomes.

Nanofibrous electrospun scaffold doped with hydroxyapatite derived from sand lobster shell (Panulirus homarus) for bone tissue engineering.

Healing of significant segmental bone defects remains a challenge, and various studies attempt to make materials that mimic bone structures and have biocompatibility, bioactivity, biodegradability, and osteoconductivity to native bone tissues. In this work, a nanofiber scaffold membrane of polyvinyl alcohol (PVA)/polyvinylpyrrolidone (PVP)/chitosan (CS) combined with hydroxyapatite (HAp) from sand lobster (SL; Panulirus homarus) shells, as a calcium source, was successfully synthesized to mimic the nanoscale extracellular matrix (ECM) in the native bone. The HAp from SL shells was synthesized by co-precipitation method with Ca/P of 1.67 and incorporated into the nanofiber membrane PVA/PVP/CS synthesized by the electrospinning method with varying concentrations, i.e. 0, 1, 3, and 5% (w/v). Based on the morphological and physicochemical analysis, the addition of HAp into the nanofiber successfully showed incorporation into the nanofiber with small agglomeration at HAp concentrations of 1, 3, and 5% (w/v). This led to a smaller fiber diameter with higher concentration of Hap, and incorporating HAp into the nanofiber could improve the mechanical properties of the nanofiber closer to the trabecula bone. Moreover, in general, swelling due to water absorption increases due to higher hydrophilicity at higher HAp concentrations and leads to the improvement of the degradation process and protein adsorption of the nanofiber. Biomineralization in a simulated body fluid (SBF) solution confirms that the HAp in the nanofiber increases bioactivity, and it can be seen that more apatite is formed during longer immersion in the SBF solution. The nanofiber PVA/PVP/CS HAp 5% has the most potential for osteoblast (MC3T3E1) cell viability after being incubated for 24 h, and it allowed the cell to attach and proliferate. Additionally, the higher HAp concentration in the nanofiber scaffold membrane can significantly promote the osteogenic differentiation of MC3T3E1 cells. Overall, the PVA/PVP/CS/HAp 5% nanofiber scaffold membrane has the most potential for bone tissue engineering.

Utilizing Proteomic Analysis to Assess the Biocompatibility and Growth Behavior of Human Immunodeficiency Virus-Associated Colorectal Cancer Cells on Polylactic Acid/Polyglycolic Acid Composite Scaffold

This research aimed to analyze the biocompatibility and growth behavior of human immunodeficiency virus (HIV)-associated colorectal cancer (CRC) cells on a polylactic acid/polyglycolic acid (PLA-PCL) composite nanofiber scaffold (CNS) using proteomic analysis. The PLA-PCL CNS was prepared using a gel extraction phase separation (GEPS) method in a dioxane/ethanol solvent system, and the effects of gel temperature, PLA-PCL mass ratio (MR), solvent-to-polymer MR on the structure and properties of the PLA-PCL CNS were analyzed. Subsequently, Lappaol F (LAF) was incorporated into the PLA-PCL CNS using electrospinning (ES) technology, and tissue specimens were obtained from HIV-associated CRC patients to investigate the impacts of LAF-PLA-PCL CNS on the growth behavior of HIV-associated CRC cells through mass spectrometry and bioinformatics analysis. Field emission scanning electron microscopy (FE-SEM) revealed that the CNS could be obtained at gel temperatures between −20 °C and −10 °C, PLA-to-PCL MRs of 1:1, 3:2, 7:3, and ethanol content of 5%–15%. X-ray diffraction (XRD) and differential scanning calorimetry (DSC) showed that the PLA-to-PCL MR of 70:30 exhibited compatibility and crystallinity of the PLA-PCL CNS, while the porosity increased with an increase in PCL content. Fourier-transform infrared spectroscopy (FTIR) analysis indicated a good biological activity of the PLA-PCL CNS. The relative cell proliferation (RCP) and cytotoxicity grades exhibited no marked differences (P &gt;0.05) between the cancer cells cultured with DMEM and those with PLA-PCL scaffold extract over time. Proteomic analysis identified 127 differentially expressed proteins (DEPs) in HIV-associated CRC cells co-cultured with LAF-PLA-PCL CNS, and Gene Ontology (GO) protein and KEGG pathway enrichment analysis (KEGG analysis) revealed that the LAF-PLA-PCL NCS could affect the cell cycle (CC) of HIV-associated CRC cells. In conclusion, the PLA-PCL NCS exhibited good compatibility, crystallinity, and biological activity, while the LAF-PLA-PCL NCS showed the potential to inhibit cell proliferation by affecting the CC.

Fluocinolone Acetonide Loaded Chitosan Nanofiber Scaffolds for Treatment of Ocular Disorders: In Vitro Characterization, Ex-Vivo Corneal and Ex-Vivo Scleral Evaluation

Purpose Drugs administered in the ocular region need to overcome ocular barriers without permanently damaging the ocular tissues. Moreover, ocular disorders of the posterior segment are more difficult to treat due to invasive procedures required to reach the posterior segment. Hence, to treat posterior disorders of the eye an attempt was made to develop nanofiber (NF) scaffolds for effective management of chronic posterior uveitis. Nanofibers (NFs) were formulated using the electrospinning technique. Methods NF scaffolds were formulated using the electrospinning technique. The effect of different concentrations of chitosan on NF production was studied by considering different ratios of chitosan (CS) and polyvinyl alcohol (PVA). Physicochemical characterization of NFs was performed to evaluate developed NFs. Results The optimized NF scaffold had a diameter of 129 ± 3 nm. NF scaffolds were found to have a tensile strength of 0.2882 ± 0.078 N/m2, thickness of 0.16 ± 0.05 mm, and drug entrapment of 95 ± 2.0%. The bioadhesive strength of the NF was found to be 257.3 ± 0.04 g/cm2 indicating high bioadhesion of NFs to the ocular tissues. The in-vitro, ex-vivo corneal and ex-vivo scleral drug release after 12 h was found to be 78.4 ± 1.0%, 65.33 ± 0.2% and 78.41 ± 1.0%, respectively. Ex-vivo whole eye model experiment indicated a concentration of about 40 ± 1.75% of drug permeated from corneal layer to the vitreous humor after 12 h. The Hen’s egg test-chorioallantoic membrane study (HET-CAM) study and in-vitro cytotoxicity study on Statens Seruminstitut Rabbit Cornea (SIRC) cell lines indicated that the developed drug-loaded NF scaffolds were found to be non-toxic as compared to pure drug, thus suggesting cytocompatibility. Conclusion Results of HET-CAM, sterility and ex-vivo studies indicate that the developed formulation is non-toxic, sterile, and effective for the ocular delivery of fluocinolone acetonide to the posterior segment of eye.

Transforming layered 2D mats into multiphasic 3D nanofiber scaffolds with tailored gradient features for tissue regeneration.

Multiphasic scaffolds with tailored gradient features hold significant promise for tissue regeneration applications. Herein, this work reports the transformation of two-dimensional (2D) layered fiber mats into three dimensional (3D) multiphasic scaffolds using a 'solids-of-revolution' inspired gas-foaming expansion technology. These scaffolds feature precise control over fiber alignment, pore size, and regional structure. Manipulating nanofiber mat layers and Pluronic F127 concentrations allows further customization of pore size and fiber alignment within different scaffold regions. The cellular response to multiphasic scaffolds demonstrates the number of cells migrated and proliferated onto the scaffolds are mainly dependent on the pore size rather than fiber alignment. In vivo subcutaneous implantation of multiphasic scaffolds to rats reveals substantial cell infiltration, neo tissue formation, collagen deposition, and new vessel formation within scaffolds, greatly surpassing the capabilities of traditional nanofiber mats. Histological examination indicates the importance of optimizing pore size and fiber alignment for promotion of cell infiltration and tissue regeneration. Overall, these scaffolds have potential applications in tissue modeling, studying tissue-tissue interactions, interface tissue engineering, and high-throughput screening for optimized tissue regeneration.

Electrospun Nanofiber Scaffolds Loaded with Metal-Based Nanoparticles for Wound Healing.

Failures of wound healing have been a focus of research worldwide. With the continuous development of materials science, electrospun nanofiber scaffolds loaded with metal-based nanoparticles provide new ideas and methods for research into new tissue engineering materials due to their excellent antibacterial, anti-inflammatory, and wound healing abilities. In this review, the stages of extracellular matrix and wound healing, electrospun nanofiber scaffolds, metal-based nanoparticles, and metal-based nanoparticles supported by electrospun nanofiber scaffolds are reviewed, and their characteristics and applications are introduced. We discuss in detail the current research on wound healing of metal-based nanoparticles and electrospun nanofiber scaffolds loaded with metal-based nanoparticles, and we highlight the potential mechanisms and promising applications of these scaffolds for promoting wound healing.