Cellular organization of foreign grafts constructed from cultivated cells is critical to successful graft-host integration and tissue repair. This study described a novel human corneal endothelial cell (HCEC) therapeutic method, where cultivated adult HCEC sheet with uniform orientation was prepared and transplanted to a rabbit cornea. Having a correct morphology and intact barriers, the HCEC sheet was made by the temperature-modulated detachment of monolayered HCECs from thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm)-grafted surfaces and was delivered with proper polarity to the corneal posterior surface by a bioadhesive gelatin disc. Results of the in vivo studies, including the follow-up clinical observations and histological examinations, showed the laminated HCEC sheet was successfully integrated into rabbit cornea denuded with endothelial layer after the biodegradation of gelatin carrier. These data indicate the feasibility of the proposed procedure in cell therapy for corneal endothelial cell loss.
Multifunctional micelles for cancer cell targeting, distribution imaging, and anticancer drug delivery were prepared from an environmentally‐sensitive graft copolymer, poly(N‐isopropyl acrylamide‐co‐methacryl acid)‐g‐poly(D,L‐lactide) (P(NIPAAm‐co‐MAAc)‐g‐PLA), a diblock copolymer, methoxy poly(ethylene glycol)‐b‐poly(D,L‐lactide) (mPEG‐PLA) and two functionalized diblock copolymers, galactosamine‐PEG‐PLA (Gal‐PEG‐PLA) and fluorescein isothiocyanate‐PEG‐PLA (FITC‐PEG‐PLA). Anticancer drug, free base doxorubicin (Dox) was incorporated into the inner core of multifunctional micelles by dialysis. From the drug release study, a change in pH (from pH 7.4 to 5.0) deformed the structure of the inner core from that of aggregated P(NIPAAm‐co‐MAAc), causing the release of a significant quantity of doxorubicin (Dox) from multifunctional micelles. Multifunctional micelles target specific tumors by an asialoglycoprotein (HepG2 cells)‐Gal (multifunctional micelle) receptor‐mediated tumor targeting mechanism. This mechanism then causes intracellular pH changes which induce Dox release from multifunctional micelles and that micelles have strong effects on the viability of HepG2 cells and are abolished by galactose. Confocal laser scanning microscopy (CLSM) reveals a clear distribution of multifunctional micelles. With careful design and sophisticated manipulation, polymeric micelles can be widely used in cancer diagnosis, cancer targeting, and cancer therapy simultaneously.
Neural retinal transplantation has great potential for the alleviation of different degenerative and hereditary retinal disorders. However, because of the fragile and soft nature of retina, retinal-sheet transplantation is relatively difficult to achieve. To overcome this difficulty, we developed a technique for lamellar tissue transplantation. Biodegradable gelatin membranes were fabricated into a sandwich and encapsulated retinal grafts for transplantation. Before transplantation, we characterized the in vivo and in vitro properties of such membranes to determine the optimal sterilization procedure, that is, a sterile membrane with suitable degradability and good mechanical properties and without cytotoxicity. Three sterilization methods were conducted, with hydrogen peroxide gas plasma (HPGP), ethylene oxide (EO), and ␥-ray irradiation (␥). The results were compared with those of a control (no disinfection). Initial studies revealed that the gelatin membranes sterilized with HPGP or EO exhibited retinal pigment epithelium (RPE) cytotoxicity, whereas the membrane sterilized by 16.6-kGy ␥ ray irradiation had no RPE cytotoxicity and had enhanced mechanical properties. In the in vivo rabbit study, implanted gelatin membranes demonstrated satisfactory biocompatibility without any inflammation. Transplanted retinal sheets survived well and developed laminar structures. Such a method using gelatin membranes for tissue transportation has great potential for future routine retinal-sheet transplantation.
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