Thymosin Beta4Chitosan Nanoparticle Loaded Fibrin-binding Amniotic Membrane For Ophthalmic Delivery: Preparation, Characterization And Effect On Corneal Wound Healing

Purpose: to produce one new bio-composite materials, double releaselayer, Thymosin β4/chitosan nanoparticles crosslinking amnioticmembrane (Tβ4/CS-FBAM). Here we study the release properties of rabbitcorneal trauma in vitro via regulate the expression of the nucleartranscription factor NF-κB role in repair of the cornea.Method: Four stepsFirst: By means of the technological of ionic cross-linking toprepared the thymosin β4/chitosan nanoparticles, while to study a seriousof traits including: particle size, zeta potential, encapsulation efficiencycharacteristics.Second: ocular surface toxicity test via eye drops of thymosinβ4/chitosan nanoparticles suspension.Third: to prepared of thymosin β4/chitosan nanoparticles glueassociated amnion (Tβ4/CS-FBAM), and to investigated the traits ofthymosin β4/chitosan nanoparticles and the release performance ofthymosin β4/chitosan via nanoparticles cross-linking amniotic.Fourth: to produced rabbit corneal trauma model, by virtue ofreal-time fluorescence quantitative PCR method to detect thymosin β4/the chitosan nanoparticles plastic linked amniotic, and to explore the role ofpostoperative rabbit cornea of NF-κB its cornea-repair mechanism.Result:First of all, The thymosin β4/chitosan nanoparticles are presented asspherical, smooth surface, uniform size, particle size of354±1.6nm, thezeta potential of22.5±0.7mv, encapsulation efficiency was (76.15±2.80)%, stable system, could be released. Second, there are not anyoculatoxicity effects observed on the eye drops of Thymosin β4/chitosannanoparticles suspension. Third, we also observed that fibrin gluecontaining the nano drug were connect with amniotic membrane closely,not easy to curl. Moreover, the its surface presented as reticular structure,nano-medicine is full of them. Tβ4/CS-FBAM drug release up to14d, invitro. Finally, the observation within rabbit corneal trauma model, weobserved that the Tβ4/CS-FBAM the group, FBAM group, and Tβ4/CS ofgroup in reducing inflammation reaction, reduce the expression of NF-κBaspects are significantly better than the control group, and at each timepoint, the difference was statistically significance (P <0.05) after operation;Meanwhile, the expression of NF-κB in Tβ4/CS-FBAM group wassignificantly below Tβ4/CS group during14days,21days, and28dayspost operation(P <0.05).Conclusion: This advanced biological graft materials, Tβ4/CS-FBAM,could be employed in local slow release of thymosin β4, which act viadown regulating the expression of NF-κB in the wounds in the cornea ofrabbits to promote the corneal reparation.