Chemical, Physical, and Biological Corneal Decellularization Methods: A Review of Literature

Table 2

The summary of physical corneal decellularization methods discussed in this review.

Physical decellularization methods	Advantages/disadvantages	Significance for future research	References
Freeze-thaw cycles	(i) Good corneal decellularization effect by lysing cells; (ii) Need to be used in combination with other decellularization techniques to remove cells; (iii) Disruption to ECM structure; (iv) Better optical properties of decellularized corneal tissue compared to the use of chemical detergents	(i) Medium	[8, 27, 59]
Hypertonic and hypotonic solutions (mostly NaCl)	(i) Efficient decellularization achieved when in combination with other decellularization method (for example, with nuclease or Triton X-100); (ii) Minimal disruption in the ECM structure; (iii) Good optical properties; (iv) Less sufficient compared to chemical detergents	(i) Medium	[33, 36, 37, 39, 42–44, 60, 61]
High or ultra-high hydrostatic pressure (HHP, UHHP)	(i) Efficient decellularization; (ii) Minimal changes in the ECM structure; (iii) Solitary application; (iv) Expensive equipment required	(i) Medium	[51, 62, 63]
ScCO2	(i) Effective corneal decellularization; (ii) Reduces decellularization time; (iii) Can cause changes in ECM structure (increase gap between collagen fibrils); (iv) Can be simultaneously used to sterilize corneal tissue; (v) Less expensive compared to HHP; (vi) Limitation is the need of the complex scCO2 reactor system	(i) Medium/high	[44, 64–66]