Page 400 - IJB-9-5

International Journal of Bioprinting A sturgeon cartilage extracellular matrix-derived bioactive bioink

Figure 2. Preparation and characterization of methacrylate-modified decellularized sturgeon cartilage ECM (dSC-ECMMA). (A) Preparation of dSC-
ECM by decellularization and methacrylate-modified procedures; (B) histological analyses of native and decellularized sturgeon cartilage ECM by H&E,
SO, and Masson. Abbreviations: H&E, hematoxylin and eosin; Masson, Masson’s trichrome; SO, safranin-O.

differences between groups were calculated by one-way the decellularization process effectively eliminated resident
analysis of variance (ANOVA) at a confidence interval of chondrocytes (Figure 2B). However, obtained dSC-ECM
95% via GraphPad Prism 7.00 software. The differences required further modification, by which it could crosslink
were considered significant at p<0.05(*), p<0.01(**), and with other components of bioinks and solidify into
[23]
p<0.001(***). hydrogel during 3D bioprinting procedure . To achieve
this purpose, methacrylate (MA) was used to modify
3. Results dSC-ECM and generated methacrylate-modified dSC-
ECM (dSC-ECMMA) particles, which were theoretically
3.1. Characterization of dSC-ECM hydrogel samples photocurable and capable of photo crosslinking. After
It has been suggested that the advantages of decellularization 10 s of blue light (405 nm,10mw/cm ) exposure, the dSC-
2
include maximal clearance of cellular and genetic molecules ECMMA solutions transformed to crosslinked hydrogels.
and minimal loss of ECM components. Decellularized
cartilage extracellular matrix (Dc-ECM) seems to be an To confirm the methacrylation of dSC-ECM and
ideal natural material with bioactivity for cartilage tissue sericin, HNMR spectra tests were made to assess the
1
regeneration compared with synthetic biomaterials. In prepared samples. According to the results of H NMR
1
this study, sturgeon cartilage was prepared into dSC- spectra, it confirmed that MA was successfully conjugated
ECMMA by the procedures including tissue preparation, onto sericin as the new peaks occur at “C=C” (δ 5.6 ppm
decellularization, solubilization, and methacrylation and 6.1 ppm) and “-CH ” (δ 1.8 ppm). Furthermore, these
3
(Figure 2A). Decellularized sturgeon cartilage fragments 1 H NMR spectra results displayed that MA was successfully
and sturgeon cartilage tissues were sectioned and evaluated conjugated onto dSC-ECM as the new peaks occur at
using HE and SO staining, confirming the elimination of “C=C” (δ 5.7 ppm and 6.2 ppm) and “-CH ” (δ 1.8 ppm).
3
cellular and genetic molecules. No positive cell nuclei image Hence, the C=C bond was introduced to the dSC-ECM and
was observed in decellularized sturgeon cartilage fragments sericin respectively, by grafting MA to them and obtaining
after a series of decellularization procedures, showing that photocurable dSC-ECMMA and SerMA (Figure 3A).

Volume 9 Issue 5 (2023) 392 https://doi.org/10.18063/ijb.768