Rapidly improvement of corneal recovery by an injectable hydrogel from acellular porcine corneal stroma

Abstract

Purpose : Acellular extracellular matrix (ECM) derived hydrogels have been developed as injection scaffolds in the treatment of wound healing of dermis, spinal cord, and urinary bladders. In this research, we optimize a clinically feasible acellular porcine corneal stroma hydrogel (APCSH) and study its physical and biocompatibility properties in vitro and in vivo.

Methods : APCSH at different concentration were characterized by biochemical analysis, optical transmittance and gelation times at 37 °C, to optimize the APCSH production method. Furthermore, after observation of its surface ultrastructure by scanning electronic microscopy (SEM), its diffusion analysis and cellular infiltration were compared to fibril gel and Tisseel. Cells extract cytotoxicity assays, live/dead assays and epithelium reconstruction assays were performed using rabbit keratinocytes and keratocytes in vitro, and in vivo corneal wound healing experiments were studied in mice to further observe the biocompatibility and proliferation properties of APCSH.

Results : The APCSH were successfully prepared at concentrations ranging from 10-30 mg/ml after 48 hours. When compared with the fibril gel and Tisseel, APCSH produced by the 20mg/ml APCS showed the best transparency and acceptable gelation time at 37 °C for clinical application. SEM showed that APCSH also has highly fibrillary structures with bigger interconnecting pores than the fibrin gel and Tisseel. This may facilitate the permeability of nutrition during the cells growth, an effect that was also demonstrated by the cell infiltration quantification assay. The APCSH showed great biocompatibility as measured in the cell extract cytotoxicity assay and viability assay in vitro. Furthermore, in vitro the epithelium reconstruction assay and in vivo mice wound healing experiment indicated that APSCH was supportive of corneal epithelial cell proliferation and corneal wound healing.

Conclusions : APCSH shows good biocompatibility compared to fibril gel and Tisseel in vitro and in vivo. Its highly fibrillary structures with bigger interconnecting pores may improve cell infiltration and support corneal wound healing. The APCSH produced at 20 mg/ml showed acceptable gelation time for clinic application and may provide a potential therapeutic treatment to treat corneal diseases.

This is a 2020 ARVO Annual Meeting abstract.