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Purposes: (1) Research phospholipase A2 the joint liquid nitrogen hypoxia the frozen prepared acellular porcine corneal stroma as a scaffold for the tissue engineering corneal feasibility. (2) To observe the biological function of autologous platelet-rich plasma on cultured corneal stromal cells. (3) the feasibility study of platelet-rich gel for tissue engineering corneal stroma build. Square - law: (1) use of phospholipase A2 and liquid nitrogen hypoxia freezing process normal pig corneal tissue morphology, thickness to prepare acellular porcine corneal stroma (Acellular porcine corneal stroma, APCS), and prepared / APCS water absorption, transmittance detection, detection of mechanical properties. (2) centrifugation rabbit whole blood platelet-rich plasma (platelet-rich plasma, PRP) and platelet poor plasma (platelet-poor plasma, PPP), using different concentrations of PRP role in corneal stromal cells, CCK-8 method detection PRP to promote the role of the corneal stromal cells. (3) take the PRP with thrombin activator preparation of platelet-rich gel in a 9:1 ratio, the preparation of PRP gel tissue morphology, the role of the corneal stromal cells, adhesion, biocompatibility and many other aspects of observation and evaluation. Results: (1) of PLA2 digestion and the joint liquid nitrogen hypoxia frozen prepared APCS is highly transparent, acellular completely, ultrastructure remained intact, in addition to water absorption trends over time and the NPC, the difference was statistically significant (P lt ; 0.05), corneal thickness, transmittance, mechanical properties compared with the NPC, there was no significant difference (P gt; 0.05). (2) PRP, when the volume fraction of 2.5% -10.0%, could promote proliferation of the stromal cells of the rabbit cornea, wherein the volume fraction of 5.0%, the strongest effect. (3) Preparation of platelet-rich gel was transparent; histological structure very similar to normal corneal stroma structure, was ordered structure of porous; observed under inverted microscope and HE staining results showed corneal stromal cells in PRP gel to good adhesion, growth, proliferation and activation;, PRP gel as adhesive with pig corneal stroma adhesion to form a tight connection between the HE staining showed. Conclusions: (1) phospholipase A2 joint liquid nitrogen hypoxia FREEZING METHOD of APCS, biological and physical characteristics similar to normal cornea, it is a good scaffold for tissue engineering corneal construct a suitable. (2) PRP is effective in promoting corneal stromal cell proliferation and activation, is conducive to the rapid repair of corneal injury, to prevent the occurrence of serious complications. (3) the PRP with the activating agent prepared in the appropriate ratio of the platelet-rich gel has a high degree of transparency, orderly tissue structure, good biocompatibility, low immunogenicity, good adhesion, suitable for Construction of tissue engineering corneal stroma.
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