Lifeink® 400 Kit

Methacrylated Hyaluronic Acid

Catalog Number: #5221-1KIT

Lifeink® 400 contains 100 mg of methacrylated hyaluronic acid which has been highly purified, lyophilized and sterility tested. 

Lifeink® 400 can be solubilized at various concentrations, and UV-crosslinked to provide increasing degrees of gel stiffness.

Price: $230

Kit Contents:

  1. 100 mg of lyophilized methacrylated hyaluronic acid bioink (1/kit)

  2. Syringe couplers (2/kit)

  3. Syringe caps (2/kit)

  4. Sterile 10 mL syringe (2/kit)

  5. Irgacure 2959

Not bioprinting? Lifeink® 400 is also available as a reagent (PhotoHA®) for 3D hydrogels and cell culture. Click here to visit the PhotoHA® page.

Lifeink® 400 is a low viscosity, yet tunable biomaterial that is highly crosslinkable, and enhances shear thinning when mixed with other proteins. 

Methacrylated HA

UV Crosslinked Methacrylated Hyaluronic Acid

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Lifeink® 200 Mixed with Lifeink® 400 (PhotoHA®)

Lifeink® 400 Cell Culture

Cell viability: 95.5 ± 1.8%

Two representative images of encapsulated hMSCs in 1 wt% PhotoHA® hydrogels after 24 hours

Human mesenchymal stem cells (20 x 10^6 / mL) were encapsulated in 50 μL hydrogels (~ 4.7 mm x 2 mm). Hydrogels (1 wt% PhotoHA® were fabricated with 0.05 wt% Irgacure 2959 and exposure to 2 mW/cm2 light (320-390 nm) for 10 minutes.  After 24 hours, encapsulated cells were stained with calcein AM and ethidium homodimer and subsequently imaged on a Leica SP5 confocal microscope (using FITC/TRITC sequential scans).

Two representative images of encapsulated neural stem cells in 1 wt% PhotoHA® hydrogels after 7 days

Neural stem cells with a gfp knock-in were encapsulated in 1% Lifeink® 400 methacrylated hyaluronic acid. 0.25% LAP was added, and then the HA was polymerized by exposure to 365nm light for 1 minute, forming hydrogels ~100-200μm thick.  Images taken after day 7 at 10X magnification. The left image is phase contrast and the right is fluorescence.
Neural stem cell images courtesy of Jonathan Ramirez from Dr. Deepak Lamba’s Lab at Buck Institute.

Rheological Properties

Reaction Behavior

Rheological time sweeps (AR2000 stress controlled rheometer, TA Instruments; 0.5% strain, 1 Hz, 25°C) of PhotoHA® crosslinking with exposure to UV light (=320-390 nm) and in the presence of 0.05 wt% Irgacure 2959 (I2959). After 1 minute, the macromer solution (i.e. PhotoHA® and I2959) was exposed to UV light, resulting in the plateau of moduli before 5 minutes.

Compressive Modulus

Dynamic mechanical analysis (Q800, TA Instruments) was performed on 50 μL hydrogels fabricated in 4.7 mm diameter molds. Hydrogels were secured within a fluid cup via a 0.01 N pre-load and compressed to 30% strain at a rate of 0.5 N min-1. The Young’s modulus of each hydrogel was calculated as the slope of generated stress-strain curves between 10% and 20% strain.

PhotoHA® Concentration (wt%)

Swelling Test

50 μL hydrogels fabricated in 4.7 mm diameter molds were imaged before and after incubation in phosphate buffered saline at 25°C for 24 hours. The diameter of hydrogels were quantified using ImageJ software. 

Print Properties of Lifeink® 400

  • Native extracellular matrix

  • Printable between 4-25°C

  • Mechanical construct strength

  • Physiological pH and salt concentration

  • Sterility tested

  • UV-crosslinkable

  • Solubilize in a wide range of concentrations

  • Easily mixes with other inks to improve shear thinning and crosslinking

Product Specifications

Our product specifications for Lifeink® 400 ensure lot-to-lot consistency, and reliable and reproducible results. 

  • Appearance                              lyophilized powder

  • Package Size                            100 mg

  • pH                                               6.8 - 7.4 when solubilized in                                                         1X PBS

  • Osmolality                                 200-400 mOsmo H20/kg when                                                      solubilized in 1X PBS

  • Sterility                                      No growth

  • Crosslinking                              UV Light

  • Molecular Weight                      100-150 kDa

  • Grafting Efficiency                    ~50-70%

Want to modify Lifeink® 400?

Here are some helpful application notes:

Mixing Lifeink® 200 with Lifeink® 400

This protocol teaches how to mix our type I collagen with methacrylated hyaluronic acid in order to create tissues that more closely resemble the human body, and can be UV/Visible light crosslinked for additional strength.

For additional information such as directions for use, SDS, material origin, or certificate of analysis, click here.

Hyaluronic acid (HA) methacrylate can be used to form hydrogels for ex vivo engineering of autologous cartilage tissue[1] or as a mesenchymal stem cell carrier in cartilage repair[2]. Because the stiffness can be widely adjusted by altering concentration or UV-light exposure, methacrylated HA has been used to measure the effects of matrix stiffness on cell phenotype and function[3][4].

 

Methacrylated HA can be used for 3D bioprinting (extrusion[5], inkjet[5] and photolithographic[6]) to create structures that promote osteogenic differentiation of MSC’s[7]. The high tunability of hyaluronic acid methacrylate allows it to be mixed with, and reinforce other types of hydrogels (such as collagen, or gelatin methacrylate)[8].

 

  1. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5717235/

  2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627486/

  3. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5541838/

  4. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5447944/

  5. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615317/

  6. https://www.ncbi.nlm.nih.gov/pubmed/21773726/

  7. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460858/

  8. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5748291/

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