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. 2023;15(2):5-16.
doi: 10.17691/stm2023.15.2.01. Epub 2023 Mar 29.

Effect of Collagen and GelMA on Preservation of the Costal Chondrocytes' Phenotype in a Scaffold in vivo

E V Isaeva  1 , A A Kisel  2 , E E Beketov  3 , G A Demyashkin  4 , N D Yakovleva  5 , T S Lagoda  6 , N V Arguchinskaya  7 , D S Baranovsky  8 , S A Ivanov  9 , P V Shegay  10 , A D Kaprin  11
Affiliations

Effect of Collagen and GelMA on Preservation of the Costal Chondrocytes' Phenotype in a Scaffold in vivo

E V Isaeva et al. Sovrem Tekhnologii Med. 2023.

Abstract

The aim of the study was to compare type I collagen-based and methacryloyl gelatin-based (GelMA) hydrogels by their ability to form hyaline cartilage in animals after subcutaneous implantation of scaffolds.

Materials and methods: Chondrocytes were isolated from the costal cartilage of newborn rats using 0.15% collagenase solution in DMEM. The cells was characterized by glycosaminoglycan staining with alcian blue. Chondrocyte scaffolds were obtained from 4% type I porcine atelocollagen and 10% GelMA by micromolding and then implanted subcutaneously into the withers of two groups of Wistar rats. Histological and immunohistochemical studies were performed on days 12 and 26 after implantation. Tissue samples were stained with hematoxylin and eosin, alcian blue; type I and type II collagens were identified by the corresponding antibodies.

Results: The implanted scaffolds induced a moderate inflammatory response in both groups when implanted in animals. By day 26 after implantation, both collagen and GelMA had almost completely resorbed. Cartilage tissue formation was observed in both animal groups. The newly formed tissue was stained intensively with alcian blue, and the cells were positive for both types of collagen. Cartilage tissue was formed among muscle fibers.

Conclusion: The ability of collagen type I and GelMA hydrogels to form hyaline cartilage in animals after subcutaneous implantation of scaffolds was studied. Both collagen and GelMA contributed to formation of hyaline-like cartilage tissue type in animals, but the chondrocyte phenotype is characterized as mixed. Additional detailed studies of possible mechanisms of chondrogenesis under the influence of each of the hydrogels are needed.

Keywords: GelMA; cartilage tissue; chondrocyte phenotype; micromolding; scaffold; type I atelocollagen.

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Conflict of interest statement

Conflicts of interest. The authors claim that there are no conflicts of interest.

Figures

Figure 1.
Figure 1.. PVA mold appearance
Figure 2.
Figure 2.. Appearance of the scaffolds after the PVA mold dissolution
Figure 3.
Figure 3.. Chondrocytes of the 0th passage after 7 days of cultivation in a Petri dish
Alcian blue staining, glycosaminoglycans; vol. ×25
Figure 4.
Figure 4.. Group 1 (atelocollagen-based scaffolds); day 12 after implantation
Necrotic tissues in the implant area; staining with hematoxylin and eosin; vol. ×20. Asterisks mark the remains of the suture thread that point to the implantation site
Figure 5.
Figure 5.. Group 1 (atelocollagen-based scaffolds); day 12 after implantation
Scaffold is surrounded by a connective tissue capsule — Sc; staining with hematoxylin and eosin; vol. ×20
Figure 6.
Figure 6.. Group 1 (atelocollagen-based scaffolds); day 12 after implantation
Connective tissue capsule around the scaffold: (a) and (b) connective tissue with multinuclear cells of resorbed foreign bodies (marked with arrows on figure a–c); an asterisk indicates the remains of the suture thread at the implantation site. Staining with hematoxylin and eosin; vol. ×20; (c) is a part of figure (b); vol. ×40
Figure 7.
Figure 7.. Group 1 (atelocollagen-based scaffolds); day 12 after implantation
Mast cells in the connective tissue capsule around the implant (shown by arrows). staining with alcian blue; vol. ×40
Figure 8.
Figure 8.. Group 1 (atelocollagen-based scaffolds); day 26 after implantation
Fragments of the implant lattice (shown with arrows). Staining with hematoxylin and eosin; vol. ×20
Figure 9.
Figure 9.. Group 1 (atelocollagen-based scaffolds); day 26 after implantation
An island of cartilage tissue in the vicinity of the scaffold: (a) staining with hematoxylin and eosin; (b) staining with alcian blue; (c) immunohistochemical reaction to type II collagen; (d) immunohistochemical reaction for type I collagen; vol. ×20
Figure 10.
Figure 10.. Group 1 (atelocollagen-based scaffolds); day 26 after implantation
Brown fat (multidroplet lipocytes) in the rat subcutaneous tissue. Staining with hematoxylin and eosin; vol. ×20
Figure 11.
Figure 11.. Group 2 (GelMA-based scaffolds); day 12 after implantation
Fragments of the implant lattice in the rat subcutaneous tissue (shown with arrows). Staining with hematoxylin and eosin; vol. ×40
Figure 12.
Figure 12.. Group 2 (GelMA-based scaffolds); day 12 after implantation
Multinuclear cells of foreign bodies resorption in the connective tissue capsule (shown with arrows). Staining with hematoxylin and eosin; vol. ×40
Figure 13.
Figure 13.. Group 2 (GelMA-based scaffolds); day 12 after implantation
Cartilage tissue at the implant site: (a) staining with hematoxylin and eosin; (b) staining with alcian blue; vol. ×20
Figure 14.
Figure 14.. Group 2 (GelMA-based scaffolds); day 26 after implantation
Cartilage tissue at the implant site: (a) staining with hematoxylin and eosin; (b) staining with alcian blue; (c) immunohistochemical reaction to type II collagen; (d) immunohistochemical reaction for type I collagen; vol. ×20
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