== Effect of corneal drying on vector absorption in mice, rabbit and human corneas. 8 (AAV8) vector (1.1108genomic copies/l) expressing marker gene was used to study the effect of corneal drying on gene transfer. Animals were sacrificed on day 14 and gene expression was analyzed using commercial staining kit. Morphological changes and infiltration of inflammatory cells were examined with H & E staining and immunocytochemistry. == Results == Mice, rabbit or human corneas subjected to no or 10 s drying showed 6%8% BSS absorption whereas 20, 30, or 50 s corneal drying showed significantly high 14%19% (p<0.001), 21%22% (p<0.001), and 25%27% (p<0.001) BSS absorption, respectively. The AAV8 application on mouse cornea after 50 s drying showed significantly higher PF-06700841 tosylate transgene delivery (p<0.05) in vivo with mild-to-moderate changes in corneal morphology. The 30 s of drying also showed significantly (p<0.05) high transgene delivery in mouse stroma in vivo without jeopardizing corneal morphology whereas 10 or 20 s drying showed moderate degree of gene transfer with no altered corneal morphology. Corneas that underwent 50 s drying showed high CD11b-positive cells (p<0.01) compared to control corneas whereas 20 or 30 s air-dried corneas showed insignificant CD11b-positive cells compared to control corneas. == Conclusions == Controlled corneal drying with hair dryer increases vector absorption significantly. The dispensing of efficacious AAV serotype into cornea with optimized minimally invasive topical application technique could provide high and targeted expression of therapeutic genes in the stroma in vivo without causing significant side effects. == Introduction == Gene therapy is an attractive approach to treat ocular diseases. Preclinical corneal gene therapy studies in animal models have shown promising results [1-5]. Nonetheless, untargeted and uncontrolled gene delivery remains a major challenge. The clinical application of gene therapy for corneal diseases is contingent on delivery of the therapeutic genes in a tissue selective manner. Our central hypothesis is that vector and vector-delivery techniques regulate transgene delivery in the cornea, and a suitable combination PF-06700841 tosylate of efficacious vector and vector-delivery technique can successfully deliver therapeutic genes into keratocytes or stroma of the cornea in vivo. Among various gene transfer vectors tested for corneal gene therapy, viral vectors have emerged as favored vectors as they exhibited high transgene expression for longer duration [5-7]. We found adeno-associated virus (AAV) vectors to be highly efficient and safe for delivering foreign genes in rodent, rabbit, equine and human cornea in vitro and in vivo [5,6,8-11]. A varied degree of tissue selective tropism among AAV serotypes has also been observed for the cornea like other PF-06700841 tosylate tissues [9-12]. The AAV serotypes 5, 8, and 9 showed superior transduction efficiency for transporting genes in the SNX25 rodent or rabbit cornea in vivo among various tested AAV serotypes [10,11,13-15]. A careful literature survey revealed that the efficacy of numerous PF-06700841 tosylate vectors has been examined for corneal gene therapy but no efforts have been made to study the role of vector-delivery techniques in corneal gene transfer. The stroma constitutes 90% of the corneal tissue and its cellular components play important role in maintaining corneal transparency, function and pathology. The stroma is affected in a variety of corneal disorders such as graft rejection, haze, neovascularization, herpes keratitis, fibrosis and scarring. Gene therapy treatments without any side effects to treat stromal corneal disorders require localized expression of therapeutic genes into keratocytes and/or the stroma. For this purpose, numerous recombinant PF-06700841 tosylate viruses and lipids were tested administering variable volume, concentration, strength, duration and frequency of vector in the cornea via microinjection, topical, electroporation, ultrasound or gene gun [16]. Most of these studies are focused on identifying an optimal vector and dose for delivering genes in the cornea in vivo. To the best of our knowledge, no study harvested vector delivery-techniques as a tool and studied its role for enhancing and targeting gene delivery in the corneal.