Control of crosslinking for tailoring collagen-based scaffolds stability and mechanics

Davidenko, Natalia; Schuster, CF; Bax, Daniel V.; Raynal, Nicolas; Farndale, Richard W.; Best, Serena M.; Cameron, Ruth E.

doi:10.1016/j.actbio.2015.07.034

Cited by 211 publications

(212 citation statements)

References 52 publications

(73 reference statements)

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“…These suspensions/solution were poured into silicone rubber trays (Lakeland, UK) and lyophilised in a VirTis adVantage bench-top freeze-drier (Biopharma Process Systems, UK) using a cycle adapted from our previous work [14, 15, 30]. Temperature of −26 °C for freezing and 0 °C for drying under vacuum (less than 100 mTorr) were applied.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry

Davidenko

Schuster

Bax

et al. 2016

J Mater Sci: Mater Med

335

280

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Studies of cell attachment to collagen-based materials often ignore details of the binding mechanisms—be they integrin-mediated or non-specific. In this work, we have used collagen and gelatin-based substrates with different dimensional characteristics (monolayers, thin films and porous scaffolds) in order to establish the influence of composition, crosslinking (using carbodiimide) treatment and 2D or 3D architecture on integrin-mediated cell adhesion. By varying receptor expression, using cells with collagen-binding integrins (HT1080 and C2C12 L3 cell lines, expressing α2β1, and Rugli expressing α1β1) and a parent cell line C2C12 with gelatin-binding receptors (αvβ3 and α5β1), the nature of integrin binding sites was studied in order to explain the bioactivity of different protein formulations. We have shown that alteration of the chemical identity, conformation and availability of free binding motifs (GxOGER and RGD), resulting from addition of gelatin to collagen and crosslinking, have a profound effect on the ability of cells to adhere to these formulations. Carbodiimide crosslinking ablates integrin-dependent cell activity on both two-dimensional and three-dimensional architectures while the three-dimensional scaffold structure also leads to a high level of non-specific interactions remaining on three-dimensional samples even after a rigorous washing regime. This phenomenon, promoted by crosslinking, and attributed to cell entrapment, should be considered in any assessment of the biological activity of three-dimensional substrates. Spreading data confirm the importance of integrin-mediated cell engagement for further cell activity on collagen-based compositions. In this work, we provide a simple, but effective, means of deconvoluting the effects of chemistry and dimensional characteristics of a substrate, on the cell activity of protein-derived materials, which should assist in tailoring their biological properties for specific tissue engineering applications.Graphical Abstract

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Section: Methodsmentioning

confidence: 99%

Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry

Davidenko

Schuster

Bax

et al. 2016

J Mater Sci: Mater Med

335

280

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show abstract

“…35,37 Thus, the material density of the collagen sample and UHANWs/Col porous nanocomposite with 66.7 wt% UHANWs was calculated to be 1.30 and 2.14 g mL À1 , respectively. The apparent density of the freeze-dried porous nanocomposite was calculated using the equation (r a ¼ m/V), where m and V are the mass and volume of the freeze-dried porous nanocomposite, respectively.…”

Section: Porosity Of the As-prepared Porous Nanocompositementioning

confidence: 99%

“…22,35,36 The relative density (r a /r m ) was determined from the apparent density (r a ) and material density (r m ) of the freezedried porous nanocomposite. The material density of solid collagen and HAP was assumed to be 1.30 and 3.15 g mL À1 ,…”

Section: Porosity Of the As-prepared Porous Nanocompositementioning

confidence: 99%

Hydroxyapatite nanowire/collagen elastic porous nanocomposite and its enhanced performance in bone defect repair

Sun

Chen

2018

RSC Adv.

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The synthetic bone grafts that mimic the composition and structure of human natural bone exhibit great potential for application in bone defect repair. In this study, a biomimetic porous nanocomposite consisting of ultralong hydroxyapatite nanowires (UHANWs) and collagen (Col) with 66.7 wt% UHANWs has been prepared by the freeze drying process and subsequent chemical crosslinking. Compared with the pure collagen as a control sample, the biomimetic UHANWs/Col porous nanocomposite exhibits significantly improved mechanical properties. More significantly, the rehydrated UHANWs/Col nanocomposite exhibits an excellent elastic behavior. Moreover, the biomimetic UHANWs/Col porous nanocomposite has a good degradable performance with a sustained release of Ca and P elements, and can promote the adhesion and spreading of mesenchymal stem cells. The in vivo evaluation reveals that the biomimetic UHANWs/Col porous nanocomposite can significantly enhance bone regeneration compared with the pure collagen sample. After 12 weeks implantation, the woven bone and lamellar bone are formed throughout the entire UHANWs/Col porous nanocomposite, and connect directly with the host bone to construct a relatively normal bone marrow cavity, leading to successful osteointegration and bone reconstruction. The as-prepared biomimetic UHANWs/Col porous nanocomposite is promising for applications in various fields such as bone defect repair.

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“…A UV spectrophotometric method with the use of 2,4,6-trinitrobenzenesulfonic acid (TNBS) as a derivatizing agent with a strong UV chromophore has been applied to evaluate the extent of gelatin cross-linking (30,(56)(57)(58)(59)(60)(61)(62). This method was originally developed by Okuyama (63) and Satake et al (64) to determine the free amino groups of amino acids and peptides in column eluates.…”

Section: Ultra Violet Spectrophotometrymentioning

confidence: 99%

Dissolution of Gelatin Capsules: Evidence and Confirmation of Cross-Linking

Lu¹,

Shah²

2017

Dissolution Technol.

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Cross-linking is a common problem in the dissolution of gelatin capsules. Cross-linking is characterized by a bridge across the peptide backbone of the gelatin molecule which creates water insoluble membranes or pellicles during dissolution testing. The chemical covalent bonding between gelatin chains is typically triggered by catalytic amounts of aldehyde and/or from exposure to high temperature and humidity. Cross-linking gelatin capsules results in a slower release of the drug or no release at all in common dissolution media. If the gelatin capsule dissolution fails acceptance criteria due to evidence of cross-linking, the US Pharmacopeia allows the use of enzymes in the dissolution medium and requires twotier dissolution testing. Studies have shown that drugs from cross-linked capsules are available in vivo to the patient, justifying the use of enzymes in the in vitro dissolution test. The literature contains several examples supporting evidence of cross-linking. However, confirming the degree of cross-linking continues to be a practical challenge due to a lack of quantitative procedures for measuring the extent of cross-linking and variability in options for formulations, excipients, level and types of stresses, and the experience level of the analysts. This article reviews the methodologies to confirm the evidence of cross-linking such as visual observation, capsule shell switch test, and spectroscopic determination of cross-linking, with an aim to facilitate the discussion and establishment of an acceptable standard approach.

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Control of crosslinking for tailoring collagen-based scaffolds stability and mechanics

Abstract: Graphical abstract

Cited by 211 publications

References 52 publications

Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry

Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry

Hydroxyapatite nanowire/collagen elastic porous nanocomposite and its enhanced performance in bone defect repair

Dissolution of Gelatin Capsules: Evidence and Confirmation of Cross-Linking

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