Experimental Study On The Treatment Of The Corneal Scar By The Combination Of Minimal Invasive Surgery And Tissue Engineering

Objective: The purpose of this study was to evaluate the outcome of the part corneal scar resection via minimal invasive surgery and to determine the feasibility of the application of the keratocytes cultured in vitro to repaire the tissue defection. Method: 1. The model of rabbits corneal scar was established by alkali burn. Then the scar was cut partly by minimal invasive surgery. The outcome was recorded; 2. The primary culture and the subculture of the keratocytes were performed. The growth curve of keratocytes was made by MTT assay and the influence of the medium with or without serum on keratocytes was synchronously compared; 3. The cell suspension(1.0 X 106cell/ml) of the keratocytes was prepared and then transplanted into the normal rabbit corneal stromal. The appearance of the host corneal was observed. After 2 weeks the viability of the transplanted keratocytes was determined with simultaneous double- staining procedure using fluorescein diacetate(FDA) and propidium iodide(PI). 4. The TGF- a polybutylcyanoacrylate nanoparticles(TGF a -PBCA-NP) were produced by the method of emulsion polymerization. Both the TGF a -PBCA-NP and TGF- a were separately added into the keratocytes culture system and the influence of them was compared, as assessed by MTT assay. After 3 month's preservation, the ability of the TGF a -PBCA-NP and TGF- a were identified by the same way. Result: 1. The corneal scar was decreased after being partly cut by minimal invasive surgery. 2 The improved culture technique for keratocytes is convenient and effective. Serum can influence some characteristic of keratocytes. 3. The white spot induced by the transplantedkeratocytes in the host corneal was significantly decreased or disappeared. FDA-PI staining showed that most transplanted cells are alive but the dead cells are more than that in normal corneal(P<0.05). 4. The promoting proliferation effect of the TGF a -PBCA-NP was higher than that of TGF- a alone (P<0.05). The 3 months storing did not affect the inductive ability of the TGF a -PBCA-NP(P<0.05). Result: The rabbit corneal scar could be decreased by the minimal invasive surgery. The keratocytes to repaire the tissue defection could be expand in vitro. These cells could be alive after being transplanted into normal rabbits corneal stromal and their cellular arragement might be turned to transparent type arrangement owing to the host envirment. The TGF a -PBCA-NP was stable, workable, injectable, biocompatibie and was worth using with the cultured keratocytes to repair the corneal stromal defection,...