Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Amine-reactive crosslinking enhances type 0 collagen hydrogel properties for regenerative medicine

doi: 10.3389/fbioe.2024.1391728

Figure Lengend Snippet: (A) Schematic representation of crosslinking of RpCol with selected chemical crosslinking agents; EDC with or without NHS or sNHS, multi-arm functionalised PEG-derivatives (8-arm shown), and genipin. Adapted from ( ; ; ). Created in BioRender.com . (B) FTIR-ATR spectra of RpCol before (uncrosslinked RpCol) and after crosslinking with chemical crosslinking agents (4-arm PEG-SCM, 8-arm PEG-SCM, EDC/NHS, EDC/sNHS, and genipin) to form hydrogels.

Article Snippet: Chemical crosslinking agents were solubilised in PBS to 100 mg/mL (w/v); and for 4%, 8%, and 16% (v/v) PEG-SCM (polyethylene glycol succinimidyl carboxyl methyl ester) hydrogels (CreativePEGWorks, 4-arm (PSB-486), 8-arm (PSB-841)), 10, 20, and 40 μL of PEG-SCM, respectively, was added to 250 μL of RpCol.

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Amine-reactive crosslinking enhances type 0 collagen hydrogel properties for regenerative medicine

doi: 10.3389/fbioe.2024.1391728

Figure Lengend Snippet: Hydrolytic degradation of RpCol hydrogels formed with chemical crosslinking agents. (A–C) Multi-arm functionalised PEG derivatives at concentrations of 4%, 8%, and 16%; (A) 20 kDa 4-arm PEG-GAS, (B) 10 kDa 4-arm PEG-SCM, (C) 10 kDa 8-arm PEG-SCM. (D–I) EDC/NHS or EDC/sNHS at ratios 1:1, 2:1, and 5:1 at concentrations of (D) 1% EDC/NHS, (E) 1% EDC/sNHS, (F) 5% EDC/NHS, (G) 5% EDC/sNHS, (H) 10% EDC/NHS, (I) 10% EDC/sNHS. (J) Different concentrations of genipin; 1, 2.5, and 5 mM. (K) Most stable formulation of each crosslinker; 20 kDa 16% 4-arm PEG-GAS, 10 kDa 16% 4-arm PEG-SCM, 10 kDa 16% 8-arm PEG-SCM, 10% EDC/NHS 5:1, 10% EDC/sNHS 1:1, and 5 mM genipin. Data presented as mean ± standard error of the mean of three independent experiments performed in triplicate.

Article Snippet: Chemical crosslinking agents were solubilised in PBS to 100 mg/mL (w/v); and for 4%, 8%, and 16% (v/v) PEG-SCM (polyethylene glycol succinimidyl carboxyl methyl ester) hydrogels (CreativePEGWorks, 4-arm (PSB-486), 8-arm (PSB-841)), 10, 20, and 40 μL of PEG-SCM, respectively, was added to 250 μL of RpCol.

Techniques: Formulation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Amine-reactive crosslinking enhances type 0 collagen hydrogel properties for regenerative medicine

doi: 10.3389/fbioe.2024.1391728

Figure Lengend Snippet: (A–F) Representative SEM images at × 100 magnification of the cross-sections of RpCol before and after the addition of chemical crosslinking agents; (A) uncrosslinked RpCol, (B) 4-arm PEG-SCM, (C) 8-arm PEG-SCM, (D) EDC/NHS, (E) EDC/sNHS, and (F) genipin, to form hydrogels. (G, H) Glucose uptake of RpCol hydrogels formed with selected chemical crosslinking agents; (G) 4%, 8%, and 16% 8-arm PEG-SCM and 1%, 5%, and 10% EDC/sNHS 1:1, and (H) 16% 8-arm PEG-SCM and 5% EDC/sNHS 1:1. Data presented as mean ± standard error of the mean of two independent experiments performed in triplicate.

Article Snippet: Chemical crosslinking agents were solubilised in PBS to 100 mg/mL (w/v); and for 4%, 8%, and 16% (v/v) PEG-SCM (polyethylene glycol succinimidyl carboxyl methyl ester) hydrogels (CreativePEGWorks, 4-arm (PSB-486), 8-arm (PSB-841)), 10, 20, and 40 μL of PEG-SCM, respectively, was added to 250 μL of RpCol.

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Amine-reactive crosslinking enhances type 0 collagen hydrogel properties for regenerative medicine

doi: 10.3389/fbioe.2024.1391728

Figure Lengend Snippet: Rheological analyses of RpCol hydrogels formed with selected chemical crosslinking agents. (A, B) amplitude sweeps of RpCol hydrogels crosslinked with; (A) 16% 8-arm PEG-SCM, and (B) EDC/sNHS 5% 1:1. (C, D) time sweeps of RpCol hydrogels crosslinked with; (C) 16% 8-arm PEG-SCM, and (D) EDC/sNHS 5% 1:1.

Article Snippet: Chemical crosslinking agents were solubilised in PBS to 100 mg/mL (w/v); and for 4%, 8%, and 16% (v/v) PEG-SCM (polyethylene glycol succinimidyl carboxyl methyl ester) hydrogels (CreativePEGWorks, 4-arm (PSB-486), 8-arm (PSB-841)), 10, 20, and 40 μL of PEG-SCM, respectively, was added to 250 μL of RpCol.

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Amine-reactive crosslinking enhances type 0 collagen hydrogel properties for regenerative medicine

doi: 10.3389/fbioe.2024.1391728

Figure Lengend Snippet: Immunostaining. Representative images on day 1 (A–F) and day 3 (G–L) of the morphology (A–C) and (G–I) and viability (D–F) and (J–L) of TERT-hMSCs encapsulated within RpCol hydrogels. (A, D, G, J) = controls (cells cultured on tissue culture plastic). (B, E, H, K) = 5% EDC/sNHS 1:1-crosslinked RpCol hydrogels. (C, F, I, L) = 16% 8-arm PEG-SCM-crosslinked RpCol hydrogels. Scale bars = 150 µm.

Article Snippet: Chemical crosslinking agents were solubilised in PBS to 100 mg/mL (w/v); and for 4%, 8%, and 16% (v/v) PEG-SCM (polyethylene glycol succinimidyl carboxyl methyl ester) hydrogels (CreativePEGWorks, 4-arm (PSB-486), 8-arm (PSB-841)), 10, 20, and 40 μL of PEG-SCM, respectively, was added to 250 μL of RpCol.

Techniques: Immunostaining, Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Amine-reactive crosslinking enhances type 0 collagen hydrogel properties for regenerative medicine

doi: 10.3389/fbioe.2024.1391728

Figure Lengend Snippet: Metabolic activity and anti-inflammatory cytokine production. (A) metabolic activity of TERT-hMSCs encapsulated within RpCol hydrogels formed with selected chemical crosslinking agents (EDC/sNHS 5% 1:1 and 8-arm PEG-SCM 16%). (B) IL-10 production of TERT-hMSCs encapsulated within RpCol hydrogels formed with selected chemical crosslinking agents (EDC/sNHS 5% 1:1 and 8-arm PEG-SCM 16%). Data presented as mean ± standard error of the mean of two independent experiments performed in triplicate. Asterix represent degree of significance; p < 0.05*, p < 0.01**, p < 0.001***, p < 0.0001****. Paired t-tests were used to analyse the same chemical crosslinking agent between different days. Unpaired t-tests were used to analyse different chemical crosslinking agents between the same day. RFU = relative fluorescence units.

Article Snippet: Chemical crosslinking agents were solubilised in PBS to 100 mg/mL (w/v); and for 4%, 8%, and 16% (v/v) PEG-SCM (polyethylene glycol succinimidyl carboxyl methyl ester) hydrogels (CreativePEGWorks, 4-arm (PSB-486), 8-arm (PSB-841)), 10, 20, and 40 μL of PEG-SCM, respectively, was added to 250 μL of RpCol.

Techniques: Activity Assay, Fluorescence