Fig. 5.

Ex vivo retention and biocompatibility of GELGYM. a) Application of GELGYM as glue to attach two human corneoscleral limbal pieces (FD of 175% and 22.5% w/v and 5 min crosslinking) and their retention rate (b) as a function of incubation time (T = 37 °C and 5% CO₂). c) Representative TEM images of the cross-sectional interface of tissue-glue after 1 and 6-month incubation in culture media (the red and blue arrows show the stratified HCEp and HCF respectively (the white and black arrows illustrate GELGYM and highly organized collagenous fibers of the tissue, respectively, separated by the interface [green arrow]. The violet arrow shows newly synthesized and partially organized collagen fibers as the GELGYM is biodegraded by HCF, indicating tissue regeneration). d) Fluorescent immunostaining images of the cross-sectional interface after 6-month incubation in culture media, showing the expression (green signal) of CK 3/12 in HCEp (top), ALDH3A1 in HCF (middle), and α-SMA in HCF (bottom); all cell nuclei were counterstained using DAPI (blue). e) Representative fluorescent immunostaining image of HCEn cultured on GELGYM, after 6 days, indicating the expression of ZO-1 (pink signal). f) Complement activation by GELGYM, noted by an increased level of C3bc and soluble terminal complement complex (sC5b-9) compared to the background activation after 30 min (T30). (T0 represents the status of complement activation immediately after drawing the blood from the donor (*p < 0.05)).