Enzymatic hydrolysis of collagen by pancreatin and thrombin as a step in the formation of scaffolds

Семенычева, Л. Л.; Egorikhina, Marfa N.; Chasova, Victoria; Valetova, N. B.; Podguzkova, M. V.; Astanina, M. V.; Kuznetsova, Yu. L.

doi:10.1007/s11172-020-2738-2

Cited by 7 publications

(8 citation statements)

References 14 publications

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“…The CC digest solution contains no initial polymer with Mw ~ 300 kDa but does contain macromolecules with Mw ~ 100 kDa [ 60 ]. This Mw corresponds to the length of one α-chain; apparently, these are the products of the CC denaturation [ 33 , 34 ].…”

Section: Resultsmentioning

confidence: 99%

Changes in the Molecular Characteristics of Bovine and Marine Collagen in the Presence of Proteolytic Enzymes as a Stage Used in Scaffold Formation

et al. 2021

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Biopolymers, in particular collagen and fibrinogen, are the leading materials for use in tissue engineering. When developing technology for scaffold formation, it is important to understand the properties of the source materials as well as the mechanisms that determine the formation of the scaffold structures. Both factors influence the properties of scaffolds to a great extent. Our present work aimed to identify the features of the molecular characteristics of collagens of different species origin and the changes they undergo during the enzymatic hydrolysis used for the process of scaffold formation. For this study, we used the methods of gel-penetrating chromatography, dynamic light scattering, reading IR spectra, and scanning electron microscopy. It was found that cod collagen (CC) and bovine collagen (BC) have different initial molecular weight parameters, and that, during hydrolysis, the majority of either type of protein is hydrolyzed by the proteolytic enzymes within the first minute. The differently sourced collagen samples were also hydrolyzed with the formation of two low molecular fractions: Mw ~ 10 kDa and ~20 kDa. In the case of CC, the microstructure of the final scaffolds contained denser, closely spaced fibrillar areas, while the BC-sourced scaffolds had narrow, short fibrils composed of unbound fibers of hydrolyzed collagen in their structure.

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Section: Resultsmentioning

confidence: 99%

Changes in the Molecular Characteristics of Bovine and Marine Collagen in the Presence of Proteolytic Enzymes as a Stage Used in Scaffold Formation

et al. 2021

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“…The scaffolds were formed under enzymatic hydrolysis, thrombin being added to the starting composite in order to deconjugate the polypeptide bonds in the main structure-forming proteins-the fibrin and collagen. As this results in the formation of products of enzymatic hydrolysis of the collagen and fibrinogen [53,54], the scaffold structure is formed from these. It has been shown that Type 1 collagens of different species origin differ in molecular weight and that this determines the differences in the products of these protein hydrolyses and, ultimately, therefore, the differences in the structure of the resulting scaffolds [55,56].…”

Section: Discussionmentioning

confidence: 99%

Aspects of In Vitro Biodegradation of Hybrid Fibrin–Collagen Scaffolds

et al. 2021

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The success of the regenerative process resulting from the implantation of a scaffold or a tissue-engineered structure into damaged tissues depends on a series of factors, including, crucially, the biodegradability of the implanted materials. The selection of a scaffold with appropriate biodegradation characteristics allows for synchronization of the degradation of the construct with the processes involved in new tissue formation. Thus, it is extremely important to characterize the biodegradation properties of potential scaffold materials at the stage of in vitro studies. We have analyzed the biodegradation of hybrid fibrin–collagen scaffolds in both PBS solution and in trypsin solution and this has enabled us to describe the processes of both their passive and enzymatic degradation. It was found that the specific origin of the collagen used to form part of the hybrid scaffolds could have a significant effect on the nature of the biodegradation process. It was also established, during comparative studies of acellular scaffolds and scaffolds containing stem cells, that the cells, too, make a significant contribution to changes in the biodegradation and structural properties of such scaffolds. The study results also provided evidence indicating the dependency between the pre-cultivation period for the cellular scaffolds and the speed and extent of their subsequent biodegradation. Our discussion of results includes an attempt to explain the mechanisms of the changes found. We hope that the said results will make a significant contribution to the understanding of the processes affecting the differences in the biodegradation properties of hybrid, biopolymer, and hydrogel scaffolds.

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“…Collagen dispersion in acetic acid (Mn = 244 kDa, Mw = 279 kDa, Mz = 304 kDa, and PDI = 1.14) was obtained using the process described in [ 37 ]. The methods used to purify organic solvents and MMA were outlined in [ 38 ].…”

Section: Methodsmentioning

confidence: 99%

Scaffold Chemical Model Based on Collagen—Methyl Methacrylate Graft Copolymers

Kuznetsova,

Gushchina,

Lobanova

et al. 2023

Polymers

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Polymerization of methyl methacrylate (MMA) in aqueous collagen (Col) dispersion was studied in the presence of tributylborane (TBB) and p-quinone: 2,5-di-tert-butyl-p-benzoquinone (2,5-DTBQ), p-benzoquinone (BQ), duroquinone (DQ), and p-naphthoquinone (NQ). It was found that this system leads to the formation of a grafted cross-linked copolymer. The inhibitory effect of p-quinone determines the amount of unreacted monomer, homopolymer, and percentage of grafted poly(methyl methacrylate) (PMMA). The synthesis combines two approaches to form a grafted copolymer with a cross-linked structure—“grafting to” and “grafting from”. The resulting products exhibit biodegradation under the action of enzymes, do not have toxicity, and demonstrate a stimulating effect on cell growth. At the same time, the denaturation of collagen occurring at elevated temperatures does not impair the characteristics of copolymers. These results allow us to present the research as a scaffold chemical model. Comparison of the properties of the obtained copolymers helps to determine the optimal method for the synthesis of scaffold precursors—synthesis of a collagen and poly(methyl methacrylate) copolymer at 60 °C in a 1% acetic acid dispersion of fish collagen with a mass ratio of the components collagen:MMA:TBB:2,5-DTBQ equal to 1:1:0.015:0.25.

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Enzymatic hydrolysis of collagen by pancreatin and thrombin as a step in the formation of scaffolds

Cited by 7 publications

References 14 publications

Changes in the Molecular Characteristics of Bovine and Marine Collagen in the Presence of Proteolytic Enzymes as a Stage Used in Scaffold Formation

Changes in the Molecular Characteristics of Bovine and Marine Collagen in the Presence of Proteolytic Enzymes as a Stage Used in Scaffold Formation

Aspects of In Vitro Biodegradation of Hybrid Fibrin–Collagen Scaffolds

Scaffold Chemical Model Based on Collagen—Methyl Methacrylate Graft Copolymers

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