Fig. 2.

Preparation process of ECM hydrogel derived from demineralized and decellularized human bone. (a) Human femoral heads from haematologically normal patients, both female and male, aged 65 ± 12 years old, were cut in half, and the trabecular bones collected and fragmented. (b) The fragments underwent demineralization in 0.5N HCl for 24 h, followed by lyophilization. The lyophilised powders were then sieved through stainless steel sieves to obtain powders in the ranges of 45–250 μm, 250–1000 μm, and 1000–2000 μm. (c) After decellularization in a Trypsin/EDTA solution, the powders were digested in a pepsin solution for 3, 5, and 7 days, respectively. The supernatant from the digestion solution was mixed with 0.1N NaOH, 10 × PBS, and 1 × PBS and incubated at 37 °C for 1 h to induce gelation.