Background. This study was performed to investigate whether transplantation of bioengineered human corneal endothelial cell (HCEC) sheet grafts into corneas denuded of endothelium could restore corneal function and clarity in a rabbit model. Methods. After being labeled with PKH26 fluorescent dye, the adult HCECs derived from eye bank corneas were cultivated on the thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm)-grafted surfaces for 3 weeks at 37°C, and were harvested as transplantable cell sheets after incubation for 45 min at 20°C. Attached by gelatin hydrogel discs, the bioengineered cell monolayers were transplanted to rabbit corneas denuded of endothelium (HCEC sheet group). Traumatized rabbit corneas were served as controls. Postsurgical corneas underwent clinical observations and histological examinations for 6 months. Results. By transmission electron microscopy and Western blot analysis of zonula occludens-1 and Na⁺,K⁺-adenosine triphosphatase proteins, the structure and function of HCEC sheets resembled those of native corneal endothelium. After endothelial cells were removed, corneas of each group turned severe edematous and opaque. In the HCEC sheet groups, corneal clarity was gradually restored and corneal thickness was significantly less than that in the control groups (P<0.05). The attached PKH26-positive HCECs spread on rabbit Descemet’s membrane after receiving cell sheet grafts. Intraocular delivery of HCEC sheets by means of a minimally invasive technique (i.e., small-incision surgery using biodegradable hydrogels) demonstrated long-term graft integration with damaged corneas. Conclusions. These results indicate that using cultured HCECs and functional biomaterials, PNIPAAm and gelatin, an effective cell sheet-based therapy can be developed for the treatment of corneal endothelium deficiency.

Human corneal endothelial cells (HCECs) maintain corneal clarity by means of a barrier function and pump-leak mechanism.¹ Regarded as nonproliferative in vivo,² the number of HCECs decreases with aging and because of other factors such as trauma, inflammation, and the use of contact lenses. Full-thickness corneal transplantation (penetrating keratoplasty) is currently the most common way to treat corneas that are opacified owing to endothelial dysfunction. Because the supply of donor corneas is insufficient and because of the potential for complications in penetrating keratoplasty, use of cultured HCECs for the replacement of the endothelium alone would be a substantial advantage.

Background: Tissue-engineered solutions for promoting the tendon graft incorporation within the bone tunnel appear to be promising. Hypothesis: To determine the feasibility that conjugation of hyaluronic acid-tethered bone morphogenetic protein-2 can be used to stimulate periosteal progenitor cells direct fibrocartilagenous attachment and new bone formation in an extra-articular tendon-bone healing model. Study Design: Controlled laboratory study. Methods: A total of 42 mature New Zealand White rabbits were used. The long digitorum extensor tendon was transplanted into a bone tunnel of the proximal tibia. The tendon was pulled through a drill hole in the proximal tibia and attached to the medial aspect of the tibia. Photopolymerizable hydrogel based on poly (ethylene glycol) diacrylate with hyaluronic acid-tethered bone morphogenetic protein-2 was injected and photogelated in a bone tunnel. Histological and biomechanical examination of the tendon-bone interface was evaluated at postoperative weeks 3 and 6. Results: Histological analysis showed an interface fibrocartilage and new bone formed by photoencapsulation of bone morphogenetic protein-2 and periosteal progenitor cells at 6 weeks. Biomechanical testing revealed higher maximum pullout strength and stiffness in experimental groups with a statistically significant difference at 3 and 6 weeks after tendon transplantation. Conclusion: The healing tendon-bone interface undergoes a gradual remodeling process; it appears that photoencapsulation of bone morphogenetic protein-2 and periosteal progenitor cells possesses a powerful inductive ability between the tendon and the bone to incorporate the healing in a rabbit model. Clinical Relevance: Novel technologies, such as those described in this study, including photopolymerization and tissue engineering, may provide minimally invasive therapeutic procedures via arthroscopy to enhance biological healing after reconstruction of the anterior cruciate ligament.

The purpose of this study was to develop and evaluate thermally responsive copolymers, which contain temperature- and pH-sensitive segments that are either alternating in or grafted onto the main chain, and to exploit their temperature-sensitive properties for ophthalmic drug delivery. Accordingly, two types of thermoresponsive copolymers—a linear poly(acrylic acid-co-N-isopropylacrylamide) random copolymer (PAAc-co-PNIPAAm) and a poly(acrylic acid-g-N-isopropylacrylamide) graft copolymer (PAAc-g-PNIPAAm)—were investigated for their thermosensitive in situ gel formation and potential applications for ophthalmic drug delivery. All the PAAc-g-PNIPAAm graft copolymers, and the linear PAAc-co-PNIPAAm copolymer with low acrylic acid contents, have an LCST of 34 °C; this is close to the surface temperature of the eye and can therefore be utilized for ophthalmic drug delivery. In addition, the PAAc-g-PNIPAAm graft copolymers showed a higher water content than the linear random copolymer; this is due to the high water adsorption ability of PAAc. The drug release dynamics of [³H]-epinephrine as a model showed that the linear random copolymer has a faster drug release, while the graft copolymers showed a more sustained release profile. The Ritger–Peppas model was used to account for the release of the epinephrine diffusion exponent ‘n’ which was in between 0.5 and 0.6. The release of the drug is considered mainly dependent on diffusion but other factors cannot be excluded. We suspected that the dynamics of drug release are determined by the water adsorption ability because high water content results in the formation of a larger capillary network in the polymer matrix, which promotes drug diffusion into the copolymer. The results suggest that PAAc-g-PNIPAAm graft copolymers are potential thermosensitive in situ gel-forming materials for ophthalmic drug delivery.