Hydrophilic, Porous, Fiber-Reinforced Collagen-Based Membrane for Corneal Repair.

Collagen membrane with outstanding biocompatibility exhibits immense potential in the field of corneal repair and reconstruction, but the poor mechanical properties limit its clinical application. Polycaprolactone (PCL) is a biodegradable polymer widely explored for application in corneal reconstruction due to its excellent mechanical properties, biocompatibility, easy processability, and flexibility. In this study, a PCL/collagen composite membrane with reinforced mechanical properties is developed. The membrane has a strong composite structure with collagen by utilizing a porous and hydrophilic PCL scaffold, maintaining its integrity even after immersion. The suture retention and mechanical tests demonstrate that compared with the pure collagen membrane, the prepared membrane has a greater tensile strength and twice the modulus of elasticity. Further, the suture retention strength is improved by almost two times. In addition, the membrane remains fully intact on the implant bed in an in vitro corneal defect model. Moreover, the membrane can be tightly sutured to a rabbit corneal defect, progressively achieve epithelialization, and remain unchanged during observation. Overall, the PCL/collagen composite membrane is a promising candidate as a suturable corneal restoration material in clinical keratoplasty.

Collagen membrane loaded with doxycycline through hydroxypropyl chitosan microspheres for the early reconstruction of alkali-burned cornea.

Corneal alkali burn is one of the most devastating ophthalmic emergencies correlated with remarkable morbidity resulting in severe visual impairment. Appropriate intervention in the acute phase determines the eventual outcome for later corneal restoration treatment. Since the epithelium plays an essential role in inhibiting inflammation and promoting tissue repair, sustained anti-matrix metalloproteinases (MMPs) and pro-epithelialization are the prior remedies during the first week. In this study, a drug-loaded collagen membrane (Dox-HCM/Col) that could be sutured to overlay the burned cornea was developed to accelerate the early reconstruction. Doxycycline (Dox), a specific inhibitor of MMPs, was encapsulated in collagen membrane (Col) through hydroxypropyl chitosan microspheres (HCM) to develop Dox-HCM/Col, affording a preferable pro-epithelialization microenvironment and an in-situ controlled release. Results showed that loading HCM into Col prolonged the release time to 7 days, and Dox-HCM/Col could significantly suppress the expression of MMP-9 and -13 in vitro and in vivo. Furthermore, the membrane accelerated the corneal complete re-epithelialization and promoted early reconstruction within the first week. Overall, Dox-HCM/Col was a promising biomaterial membrane for treating alkali-burned cornea in the early stage, and our attempt may provide a clinically feasible method for the ocular surface reconstruction.

Self-Assembled Sulfated Hyaluronan Coating Modulates Transforming Growth Factor-Beta1 Penetration for Corneal Scarring Alleviation.

Corneal scarring caused by epithelial-stromal injury impairs corneal transparency and visual acuity. Excess secretion of transforming growth factor-beta 1 (TGF-ß1), which promotes wound closure, penetrates the corneal stroma via defects in the epithelial basement membrane and induces the differentiation of corneal fibroblasts to myofibroblasts, leading to scar formation. Modulating TGF-ß1 penetration might alleviate corneal scar formation and restore transparency. In this study, sulfated hyaluronan (sHA) coatings were self-assembled above wounded corneal stroma to modulate TGF-ß1 penetration, and their ability to alleviate corneal scarring was investigated. The formation of sHA coatings was rapid (within 30 s), and the high-sulfated hyaluronan coating efficiently blocked penetration by TGF-ß1 and reduced the concentration of TGF-ß1 in the corneal stroma. Further investigation showed that the ability of TGF-ß1 to induce differentiation of corneal fibroblasts into myofibroblasts was inhibited by sHA binding. Evaluation of corneal scarring with sHA coating in a rabbit model of lamellar resection indicated that a sHA (high sulfation) coating effectively reduced scar formation. Immunohistochemical staining of α-smooth muscle actin and optical coherence tomography of the anterior segment showed minimal scar tissue formation in the sHA group. This work presents a promising alternative to alleviate scarring in corneal epithelial-stromal injury.

MiRNA 24-3p-rich exosomes functionalized DEGMA-modified hyaluronic acid hydrogels for corneal epithelial healing.

The damage of corneal epithelium may lead to the formation of irreversible corneal opacities and even blindness. The migration rate of corneal epithelial cells directly affects corneal repair. Here, we explored ocu-microRNA 24-3p (miRNA 24-3p) that can promote rabbit corneal epithelial cells migration and cornea repair. Exosomes, an excellent transport carrier, were exacted from adipose derived mesenchymal stem cells for loading with miRNA 24-3p to prepare miRNA 24-3p-rich exosomes (Exos-miRNA 24-3p). It can accelerate corneal epithelial migration in vitro and in vivo. For application in cornea alkali burns, we further modified hyaluronic acid with di(ethylene glycol) monomethyl ether methacrylate (DEGMA) to obtain a thermosensitive hydrogel, also reported a thermosensitive DEGMA-modified hyaluronic acid hydrogel (THH) for the controlled release of Exos-miRNA 24-3p. It formed a highly uniform and clear thin layer on the ocular surface to resist clearance from blinking and extended the drug-ocular-epithelium contact time. The use of THH-3/Exos-miRNA 24-3p for 28 days after alkali burn injury accelerated corneal epithelial defect healing and epithelial maturation. It also reduced corneal stromal fibrosis and macrophage activation. MiRNA 24-3p-rich exosomes functionalized DEGMA-modified hyaluronic acid hydrogel as a multilevel delivery strategy has a potential use for cell-free therapy of corneal epithelial regeneration.

Biomimetic Convex Implant for Corneal Regeneration Through 3D Printing.

Blindness caused by corneal damage affects millions of people worldwide, and this number continues to rise. However, rapid epithelization and a stable epithelium process are the two biggest challenges for traditional corneal materials. These processes are related to corneal curvature, which is an important factor in determination of the corneal healing process and epithelial behavior during corneal damage. In this study, smooth 3D-printed convex corneal implants based on gelatin methacrylate and collagen are generated. As epithelium distribution and adhesion vary in different regions of the natural cornea, this work separates the surfaces into four regions and studies how cells sense topological cues on curvature. It is found that rabbit corneal epithelial cells (RCECs) seeded on steeper slope gradient surfaces on convex structures result in more aligned cell organization and tighter cell-substrate adhesion, which can also be verified through finite element simulation and signaling pathway analysis. In vivo transplantation of convex implants result in a better fit with adjacent tissue and stronger cell adhesion than flat implants, thereby accelerating corneal epithelialization and promoting collagen fibers and neural regeneration within 180 days. Taken together, printed convex corneal implants that facilitate corneal regeneration may offer a translational strategy for the treatment of corneal damage.

Sutureless transplantation using a semi-interpenetrating polymer network bioadhesive for ocular surface reconstruction.

The conjunctiva covers the largest area of ocular surface and is responsible for tear balance and clear vision. After trauma or surgery, the conjunctiva is prone to scarring and contracture. Transplantation with suture often implies numerous complications, such as inflammation, suture erosion, granuloma. And the suture needs to be removed, which means a secondary trauma. In this study, a bioadhesive hydrogel (GMO) for sutureless conjunctival transplantation was developed based on a semi-interpenetrating polymer network (sIPN) consisting of gelatin methacrylate (GelMA) and oxidized hyaluronic acid (OHA). The maximum adhesion strength was 157 ± 17 kPa, and the burst pressure was 357 ± 29 kPa, which was 15 times higher than the human intraocular pressure (IOP). GMO bioadhesive hydrogel significantly improved surgical efficiency and secured the collagen scaffold firmly to a rabbit conjunctival defect. The sutureless transplantation approach revealed the promoted tissue repair without scar. In conclusion, GMO bioadhesive may be an attractive alternative to suture for ocular surface reconstruction by avoiding suture-related complications and improving clinical outcome. STATEMENT OF SIGNIFICANCE: Conjunctival tissue is prone to scarring and contracture after trauma, and surgery with sutures often implies numerous complications. In this study, the ocular surface reconstruction was achieved by sutureless transplantation of conjunctival scaffold using bioadhesive hydrogel. The prepared GMO bioadhesive based on the semi-interpenetrating network of gelatin methacrylate (GelMA) and oxidized hyaluronic acid (OHA) had favorable adhesion and mechanical properties. The sutureless transplantation approach significantly improved the operation efficiency, avoided suture-related complications, and promoted the regeneration of conjunctiva. This study highlights the great potential of the sutureless repair strategy for clinical application in ocular surface reconstruction.

Photo-crosslinked GelMA/collagen membrane loaded with lysozyme as an antibacterial corneal implant.

Corneal transplantation is an effective treatment for corneal blindness. However, it brings risk factors for the occurrence of bacterial keratitis, which can affect the repair effect and even lead to transplantation failure. The difficulty in re-epithelialization is also a main problem faced by corneal transplantation. Herein, a collagen-GelMA composite membrane containing lysozyme (CGL) was developed as an antibacterial corneal implant to fill stromal defect and support re-epithelialization. Characterizations of physicochemical properties and in vitro biocompatibility revealed that the composite membranes have proper water content, light transmittance and mechanical strength as well as good biocompatibility. Particularly, the cell adhesion force and adhesion-related genes expression were evaluated and exhibited an improvement after the addition of GelMA. Furthermore, the formed CGL membrane could continuously release lysozyme and exhibited a bactericidal rate of 96% and 64% after 2 h and 72 h, respectively. The results demonstrated that this CGL membrane has promising application in corneal repair.

A carrageenan/agarose composite sponge and its immunomodulatory activities toward RAW264.7.

A kind of commercial hybrid carrageenan (HC)/agarose composite sponge containing κ-, µ-, ι-, and ν-carrageenan, which could turn into hydrogel and release carrageenan at human body temperature was fabricated for immune stimulation and modulation. Release behavior demonstrated that the hybrid carrageenan contained sponge was mechanically stable and could release carrageenan constantly. RT-PCR and ELISA experiments showed that the leaching liquor of the sponge could stimulate RAW264.7 from M0 state to a polarized state by secreting more anti-inflammatory factor IL-10 than pro-inflammatory ones, such as, IL-6 and TNF-α. Transwell experiments also indicated that the leaching liquor could promote the proliferation of NIH-3T3 by stimulating RAW264.7 of M0 state after 7 days. Results of particle size and intracellular concentration analyses suggested that the released carrageenan might enter into the cellular interior of RAW264.7 in the form of microgels or protein complexes. The sponge would be a promising candidate for skin wound dressing.

Fusion of Facial Expressions and EEG for Multimodal Emotion Recognition.

This paper proposes two multimodal fusion methods between brain and peripheral signals for emotion recognition. The input signals are electroencephalogram and facial expression. The stimuli are based on a subset of movie clips that correspond to four specific areas of valance-arousal emotional space (happiness, neutral, sadness, and fear). For facial expression detection, four basic emotion states (happiness, neutral, sadness, and fear) are detected by a neural network classifier. For EEG detection, four basic emotion states and three emotion intensity levels (strong, ordinary, and weak) are detected by two support vector machines (SVM) classifiers, respectively. Emotion recognition is based on two decision-level fusion methods of both EEG and facial expression detections by using a sum rule or a production rule. Twenty healthy subjects attended two experiments. The results show that the accuracies of two multimodal fusion detections are 81.25% and 82.75%, respectively, which are both higher than that of facial expression (74.38%) or EEG detection (66.88%). The combination of facial expressions and EEG information for emotion recognition compensates for their defects as single information sources.