Human SMILE-Derived Stromal Lenticule Scaffold for Regenerative Therapy: Review and Perspectives

A transparent cornea is paramount for vision. Corneal opacity is one of the leading causes of blindness. Although conventional corneal transplantation has been successful in recovering patients’ vision, the outcomes are challenged by a global lack of donor tissue availability. Bioengineered corneal tissues are gaining momentum as a new source for corneal wound healing and scar management. Extracellular matrix (ECM)-scaffold-based engineering offers a new perspective on corneal regenerative medicine. Ultrathin stromal laminar tissues obtained from lenticule-based refractive correction procedures, such as SMall Incision Lenticule Extraction (SMILE), are an accessible and novel source of collagen-rich ECM scaffolds with high mechanical strength, biocompatibility, and transparency. After customization (including decellularization), these lenticules can serve as an acellular scaffold niche to repopulate cells, including stromal keratocytes and stem cells, with functional phenotypes. The intrastromal transplantation of these cell/tissue composites can regenerate native-like corneal stromal tissue and restore corneal transparency. This review highlights the current status of ECM-scaffold-based engineering with cells, along with the development of drug and growth factor delivery systems, and elucidates the potential uses of stromal lenticule scaffolds in regenerative therapeutics.     

Chitosan/Alginate Hydrogel Dressing Loaded FGF/VE-Cadherin to Accelerate Full-Thickness Skin Regeneration and More Normal Skin Repairs

The process of full-thickness skin regeneration is complex and has many parameters involved, which makes it difficult to use a single dressing to meet the various requirements of the complete regeneration at the same time. Therefore, developing hydrogel dressings with multifunction, including tunable rheological properties and aperture, hemostatic, antibacterial and super cytocompatibility, is a desirable candidate in wound healing. In this study, a series of complex hydrogels were developed via the hydrogen bond and covalent bond between chitosan (CS) and alginate (SA). These hydrogels exhibited suitable pore size and tunable rheological properties for cell adhesion. Chitosan endowed hemostatic, antibacterial properties and great cytocompatibility and thus solved two primary problems in the early stage of the wound healing process. Moreover, the sustained cytocompatibility of the hydrogels was further investigated after adding FGF and VE-cadherin via the co-culture of L929 and EC for 12 days. The confocal 3D fluorescent images showed that the cells were spherical and tended to form multicellular spheroids, which distributed in about 40–60 μm thick hydrogels. Furthermore, the hydrogel dressings significantly accelerate defected skin turn to normal skin with proper epithelial thickness and new blood vessels and hair follicles through the histological analysis of in vivo wound healing. The findings mentioned above demonstrated that the CS/SA hydrogels with growth factors have great potential as multifunctional hydrogel dressings for full-thickness skin regeneration incorporated with hemostatic, antibacterial, sustained cytocompatibility for 3D cell culture and normal skin repairing.     

Boron-based bioactive glasses: Properties,processing, characterization and applications

The performance of many biological processes is thought to be affected by boron, and a deficiency is linked to delayed bone growth. Boron is therefore a bioactive element that is advantageous to both people and animals. Another well-known benefit of boron is that it promotes bone growth and wound healing. Glass structure, glass processing properties, biodegradability, biocompatibility, bioactivity, and cytotoxicity are all significantly impacted by the introduction of boron to bioactive glasses in a range of concentrations. According to research so far, boron based bioactive glasses (BBGs) frequently surpass silicate glasses in terms of bioactivity and potential for bone healing. Additionally, they could be employed as medication delivery systems for the treatment of infections and conditions like osteoporosis. By adding modifying ions, BBGs capacity to heal wounds or repair bone can be increased. Boron based bioactive glasses are typically synthesized via melt-quenching, although a more recent, more promising technique sol-gel processing is starting to attract interest. This review analyses the available literature to offer an in-depth overview of BBG properties, their real-world applications, challenges, and suggestions for future study.     

Multifunctional heteropolysaccharide hydrogel under photobiomodulation for accelerated wound regeneration

The present study evaluated the effect of fucoidan/alginate-polyethylene glycol-gellan gum (Fu/AL-PEG@GGH) hydrogel using low-level laser therapy (LLLT) on facilitating wound healing for wound care management. The hydrogel was fabricated and characterized to evaluate the wound healing potential. Cytotoxicity and apoptotic effects were evaluated with L929 and NIH3T3 cells. Uniform spherical sheets were observed with high thermal stability caused by porous matrixes with the increased cell viability and fast cell migration. Scar tissue was reduced by larger wound contraction with faster healing effects from the hydrogel + LLLT-treated group at day 14. The polysaccharides may promote wound healing due to the strong bonds by the physical cross-linking in hydrogel preparation. The results from hydrogel + LLLT-treated group confirmed an effective wound healing potential from the presence of high fibroblast and collagen deposition. Therefore, the combined practice of the hydrogel with LLLT may enhance a wound healing process for effective wound care applications.     

Advances in wound dressing based on electrospinning nanofibers

In recent years, there has been a significant focus on bioactive dressings suitable for treating chronic and acute wounds. Electrospinning nanofibers are considered advanced dressing options due to their high porosity and permeability to air and water, effective barrier properties against external pathogens, and excellent resemblance to the extracellular matrix for wound healing and skin regeneration. This article reviews the recent advancements in the application of electrospinning nanofibers for bioactive wound healing. The review begins with an overview of the wound healing process and electrospinning methods. It then explores the advantages and disadvantages of different synthetic and natural polymers used in the preparation of electrospinning wound dressings. The natural polymers discussed in this review include collagen, gelatin, silk fibroin, chitosan, hyaluronic acid, and sodium alginate. Additionally, the review delves into commonly used synthetic polymers like polyvinyl alcohol, polyvinyl chloride, polyethylene lactone, polylactide, and polyurethane for wound dressing applications. Furthermore, the review examines the blending of natural and synthetic polymers to create high-performance wound dressings. It also explores the incorporation of functional additives, such as antimicrobial agents, growth factors, and natural extracts, into electrospinning nanofibers to expedite wound healing and tissue repair. In conclusion, electrospinning is an emerging technology that provides unique opportunities for designing more effective wound dressings and care products.     

Evaluation of Wound Healing Activity of Salvianolic Acid B on In Vitro Experimental Model

Despite a wide range of bactericides and antiseptics, the treatment of chronic or complicated wounds is still a major challenge for modern medicine. Topical medications are the most sought-after new agents for use as treatment. The therapeutic concentration of their active substances is easy to achieve with the lowest possible burden on the patient’s body. This study assesses the effect of salvianolic acid B (Sal B) on the proliferation, migration, and production of collagen type III by fibroblasts, which are the most important processes in wound healing. The study was conducted on human gingival fibroblasts obtained from primary cell culture. The results showed that Sal B at a dose of 75 µg/mL increases the cell viability with significant stimulation of the cell migration as demonstrated in the wound healing assay, as well as an increase in the expression of collagen type III, which has great importance in the initial stages of wound scarring. The results obtained in the conducted studies and previous scientific reports on the antibacterial properties and low toxicity of Sal B indicate its high potential in wound healing.     

Cell migration and proliferation during monolayer formation and wound healing

The interaction of living cells with surfaces is important in applications of biomaterials, such as tissue engineering. Characterising and modelling the attachment, migration and proliferation of cells on materials used for tissue engineering provides valuable insight into their potential applications as well as a means of objective comparison. In this study, proliferation and migration of NIH-3T3 fibroblast cells on tissue culture plastic in vitro were quantified. The development of randomly scattered individual cells into confluent cell monolayers proceeded with cell density following a logistic growth pattern. Travelling cell wavefronts produced in a wound healing assay were modelled with a modified Fisher equation incorporating both diffusion and logistic growth. The diffusivity and growth rates thus determined could be used for comparison with cell behaviour on other surfaces or under different conditions. Cell tracking showed that the average effective velocity of cells varied inversely with cell density, supporting contact inhibition of cell movement.     

Sol–gel based fabrication and characterization of new bioactive glass–ceramic composites for dental applications

The fabrication of a new composite glass–ceramic with potential application in dental restoration was investigated. The developed material aims to modify the surface of dental ceramics creating bioactive surfaces able to improve material–cell interaction enhancing the bonding of the marginal gap between restoration and tooth. The application of the sol–gel method led to a microporous homogeneous glass–ceramic which can be applied as coating on commercial dental ceramic substrates. The microstructural, thermal, mechanical and biological properties of the fabricated coatings were studied and compared to the respective results of a previously investigated glass–ceramic composite. The material–cell interaction on these two sol–gel dental composites was studied in detail. The attachment and proliferation of both periodontal ligament and gingival fibroblast cells confirmed the bioactive behavior of the new materials and their ability to be potentially applied in dental restorations for soft tissue regeneration and sealing of the marginal gap.     

Skin Resident γδ T Cell Function and Regulation in Wound Repair

The skin is a critical barrier that protects against damage and infection. Within the epidermis and dermis reside γδ T cells that play a variety of key roles in wound healing and tissue homeostasis. Skin-resident γδ T cells require T cell receptor (TCR) ligation, costimulation, and cytokine reception to mediate keratinocyte activity and inflammatory responses at the wound site for proper wound repair. While both epidermal and dermal γδ T cells regulate inflammatory responses in wound healing, the timing and factors produced are distinct. In the absence of growth factors, cytokines, and chemokines produced by γδ T cells, wound repair is negatively impacted. This disruption in γδ T cell function is apparent in metabolic diseases such as obesity and type 2 diabetes. This review provides the current state of knowledge on skin γδ T cell activation, regulation, and function in skin homeostasis and repair in mice and humans. As we uncover more about the complex roles played by γδ T cells in wound healing, novel targets can be discovered for future clinical therapies.     

Fabrication of Arothron stellatus skin collagen film incorporated with Coccinia grandis as a durable wound construct

The wound dressing material has been fabricated with the novel Arothron stellatus fish skin collagen scaffold loaded with a bioactive extract obtained from Coccinia grandis (CPE) and drug Ciprofloxacin (D). The collagen scaffold was fabricated from the nonedible solid waste of marine origin (a cheaper source of raw material). The in vitro fluorescence staining of cells and in vivo evaluation of the fabricated scaffold exhibited enhanced cell adhesion and proliferation and indicated a more efficient wound healing paradigm than that of the control and other treated groups, respectively. The increased collagen synthesis and re-epithelialization of wound are proved to be potential wound constructs.     

Evaluation of scaffolding,inflammatory response,and wound healing support of a reverse thermal gel for myelomeningocele patching

A reverse thermal gel (RTG) is a promising patching material for in utero, minimally invasive coverage of myelomeningocele (MMC) defects. The injectable properties of the RTG brings the potential for significantly reduced surgical risks to the mother and fetus when compared to current open prenatal repair procedures. MMC patching materials require structural and wound healing support for tissue growth over the MMC defect area to allow a watertight seal to form and prevent further neural tissue exposure to the amniotic environment. Here, the previously described RTG is evaluated for the first time as a scaffold for skin cells, for long-term inflammation effects in neonatal mice, and for wound healing capability. Results show that the RTG can support the growth and survival of keratinocytes, dermal fibroblasts, and neuronal cells in vitro. Injections into neonatal mice demonstrate a regressing inflammatory response and support of normal wound healing. Together, these results demonstrate that the RTG has the necessary scaffolding and wound healing support necessary for MMC patching applications.     

10-Shogaol, an Antioxidant from Zingiber officinale for Skin Cell Proliferation and Migration Enhancer

In this work, one of Zingiber officinale components, 10-shogaol, was tested with 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, metal chelating ability, and reducing power to show antioxidant activity. 10-Shogaol promoted human normal epidermal keratinocytes and dermal fibroblasts cell growths. 10-Shogaol enhanced growth factor production in transforming growth factor-β (TGF-β), platelet derived growth factor-αβ (PDGF-αβ) and vascular endothelial growth factors (VEGF) of both cells. In the in vitro wound healing assay for 12 or 24 h, with 10-shogaol, the fibroblasts and keratinocytes migrated more rapidly than the vehicle control group. Thus, this study substantiates the target compound, 10-shogaol, as an antioxidant for human skin cell growth and a migration enhancer with potential to be a novel wound repair agent.     

Poly(L-lactide-co-caprolactone)/collagen electrospun mat: Potential for wound dressing and controlled drug delivery

Here we report a novel bioactive electrospun mat based on poly(L-lactide-co-caprolactone) (PLLC) and collagen for wound dressing and sustained drug delivery of gentamicin. PLLC/collagen electrospun mat loaded with 10% gentamicin showed bioactivity for 15 days against Gram-positive and Gram-negative bacteria. The in vitro cell culture of 3T3 fibroblasts confirmed that these electrospun mat provide an increased specific interface area and hydrophilicity to enhance cell attachment, proliferation, and migration. The modified PLLC/collagen mat provided an excellent enhancement in properties of antibacterial wound dressings with a minimum in vitro toxicity and high potency for promoting wound healing stages.     

Epidermal Stem Cells in Orthopaedic Regenerative Medicine

In the last decade, great advances have been made in epidermal stem cell studies at the cellular and molecular level. These studies reported various subpopulations and differentiations existing in the epidermal stem cell. Although controversies and unknown issues remain, epidermal stem cells possess an immune-privileged property in transplantation together with easy accessibility, which is favorable for future clinical application. In this review, we will summarize the biological characteristics of epidermal stem cells, and their potential in orthopedic regenerative medicine. Epidermal stem cells play a critical role via cell replacement, and demonstrate significant translational potential in the treatment of orthopedic injuries and diseases, including treatment for wound healing, peripheral nerve and spinal cord injury, and even muscle and bone remodeling.     

Fabrication of LaCl3‐containing nanofiber scaffolds and their application in skin wound healing

Poly(?‐caprolactone) (PCL)/gelatin (GE) nanofiber scaffolds with varying concentrations of lanthanum chloride (LaCl₃, from 0 to 25 mM) were fabricated by electrospinning. The scaffolds were characterized by scanning electron microscopy, contact angle and porosity measurements, mechanical strength tests, and in vitro degradation studies. In vitro cytotoxicity and cell adhesion and proliferation studies were performed to assess the biocompatibility of the scaffolds, and in vivo wound healing studies were conducted to assess scaffold applications in the clinic. All prepared scaffolds were noncytotoxic, and the growth of adipose tissue–derived stem cells on LaCl₃‐containing scaffolds was better than on the pure PCL/GE scaffold. Cell proliferation studies showed the greatest cell growth in the PCL/GE/LaCl₃ scaffolds. Further, in vivo studies proved that the PCL/GE/LaCl₃ scaffolds can promote wound healing. The results suggest that nanofiber scaffolds containing LaCl₃ promote cell proliferation and have good biocompatibility, and thus potential for application in the treatment of skin wounds. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 46672.     

Improving the performance of carbon/graphite composites through the synergistic effect of electrostatic self-assembled carbon nanotubes and nano carbon black

Superior performance fillers are considered as an effective means to enhance the performance of carbon/graphite composites. However, poor interfacial properties and incomplete filler networks limit the performance enhancement of the composites. In this study, a new method was proposed to weaken this impact through the synergistic effect of the electrostatic self-assembly of nano carbon black (NCB) onto carbon nanotubes (CNTs). The results showed that the synergistic effect between NCB and the CNTs significantly improved the mechanical and electrical properties of the composites. NCB reduces the porosity of the composites and increases the interaction between the CNTs and matrix. The compressive strength of the composite was 143.2 Mpa, and the flexural strength was 46.3 MPa, which is 210% higher than that of the pristine carbon/graphite composites. Moreover, NCB and CNTs form a globally connected synergistic network in the carbon skeleton. Composites filled with CNTs/NCB exhibited the lowest resistivity and highest thermal conductivity, with a resistance that was 42% lower than that of pristine carbon/graphite composites at 44.8 μΩ m. All of these results suggest that the synergistic effect of CNTs/NCB show great potential to improve the performance of carbon/graphite composites.     

Characterization of a Human Platelet Lysate-Loaded Keratin Hydrogel for Wound Healing Applications In Vitro

One of the promising approaches to facilitate healing and regenerative capacity includes the application of growth-factor-loaded biomaterials. Human platelet lysate (hPL) derived from platelet-rich plasma through a freeze-thaw process has been used as a growth factor rich therapeutic in many regenerative applications. To provide sustained local delivery of the hPL-derived growth factors such as epidermal growth factor (EGF), the hPL can be loaded into biomaterials that do not degrade rapidly in vivo. Keratin (KSO), a strong filamentous protein found in human hair, when formulated as a hydrogel, is shown to sustain the release of drugs and promote wound healing. In the current study, we created a KSO biomaterial that spontaneously forms a hydrogel when rehydrated with hPL that is capable of controlled and sustained release of pro-regenerative molecules. Our study demonstrates that the release of hPL is controlled by changing the KSO hydrogel and hPL-loading concentrations, with hPL loading concentrations having a greater effect in changing release profiles. In addition, the 15% KSO concentration proved to form a stable hydrogel, and supported cell proliferation over 3 days without cytotoxic effects in vitro. The hPL-loaded keratin hydrogels show promise in potential applications for wound healing with the sustained release of pro-regenerative growth factors with easy tailoring of hydrogel properties.     

Microstructure and thermophysical properties of graphite foam/glass composites

Mesophase pitch based graphite foams with different thermal properties and cell structures were infiltrated with glass by pressureless infiltration to prepare potential alternative composites for cooling electronics. Microstructure, thermal diffusivity and coefficient of thermal expansion (CTE) of the obtained composites were investigated. It was demonstrated that there was excellent wettability of the graphite foam by molten glass, and the foam framework was retained well after infiltration, which could facilitate good heat transfer throughout the composites. The highest thermal diffusivity of the composites reached 202.80 mm²/s with a density of 3.81 g/cm³. And its CTE value was 4.53 ppm/K, much lower than the corresponding calculated result (7.46 ppm/K) based on a simple “rule of mixtures” without considering the space limitations of the graphite foams. Thus, the mechanical interlocking within the space limitations of the graphite network played a crucial role in limiting the thermal expansion of the glass. The CTEs of the graphite foam/glass composites varied from 4.53 to 7.40 ppm/K depending on the graphite foam density which varied from 0.82 to 0.48 g/cm³. The CTEs were a good match to those of semiconductor chips and packaging materials.     

Silver-doped sol-gel borate glasses: Dose-dependent effect on Pseudomonas aeruginosa biofilms and keratinocyte function

Borate-based glasses have generated great interest for wound healing applications due to their ability to incorporate biologically therapeutic ions which can then be released at the site of repair attributable to their high dissolution rates. In this study, the anti-bacterial activity and cytocompatibility of sol-gel–derived silver-doped borate glasses (AgBGs) of the compositional range (60)B₂O₃–(36)CaO–(4–x)P₂O₅–(x)Ag₂O, where x = 0.0, 0.3, 0.5, and 1 (mol.%) were investigated, in vitro. The dose-dependent anti-bacterial activity of AgBGs was demonstrated against Pseudomonas aeruginosa under both planktonic conditions and pre-formed biofilms, with up to 99.7% reduction in bacterial cell counts. Lower concentrations of ionic dissolution products from AgBGs were non-toxic to keratinocytes, stimulating their growth and metabolic activity. Furthermore, compositions containing 0.3 and 0.5 mol.% Ag significantly accelerated the migration of keratinocytes at two different concentrations in an in vitro 2D wound healing model. In summary, these therapeutic AgBGs have demonstrated potential for accelerated wound healing.