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A fibrin/hyaluronic acid hydrogel for the delivery of mesenchymal stem cells and potential for articular cartilage repair.

Snyder TN, Madhavan K, Intrator M, Dregalla RC, Park D - J Biol Eng (2014)

Bottom Line: This chondrogenic hydrogel system can be delivered in a minimally invasive manner through a small gauge needle, forming a three-dimensional (3D) network structure in situ.However, an ongoing problem with fibrin/HA-based biomaterials is poor mechanical strength.Quantitative polymerase chain reaction (qPCR) of BMSCs incubated in the fibrin/HA-MA hydrogel confirmed decreased expression of collagen type 1 alpha 1 mRNA with an increase in Sox9 mRNA expression especially in the presence of a platelet lysate, suggesting early chondrogenesis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Bioengineering Department, University of Colorado, Anschutz Medical Campus, Mail Stop 8607, 12700 East 19th Avenue, Aurora, CO 80045, USA ; Regenerative Sciences, 403 Summit Blvd, Suite 201, Broomfield, CO 80021, USA.

ABSTRACT

Background: Osteoarthritis (OA) is a degenerative joint disease affecting approximately 27 million Americans, and even more worldwide. OA is characterized by degeneration of subchondral bone and articular cartilage. In this study, a chondrogenic fibrin/hyaluronic acid (HA)-based hydrogel seeded with bone marrow-derived mesenchymal stem cells (BMSCs) was investigated as a method of regenerating these tissues for OA therapy. This chondrogenic hydrogel system can be delivered in a minimally invasive manner through a small gauge needle, forming a three-dimensional (3D) network structure in situ. However, an ongoing problem with fibrin/HA-based biomaterials is poor mechanical strength. This was addressed by modifying HA with methacrylic anhydride (MA) (HA-MA), which reinforces the fibrin gel, thereby improving mechanical properties. In this study, a range of fibrinogen (the fibrin precursor) and HA-MA concentrations were explored to determine optimal conditions for increased mechanical strength, BMSC proliferation, and chondrogenesis potential in vitro.

Results: Increased mechanical strength was achieved by HA-MA reinforcement within fibrin hydrogels, and was directly correlated with increasing HA-MA concentration. Live/dead staining and metabolic assays confirmed that the crosslinked fibrin/HA-MA hydrogels provided a suitable 3D environment for BMSC proliferation. Quantitative polymerase chain reaction (qPCR) of BMSCs incubated in the fibrin/HA-MA hydrogel confirmed decreased expression of collagen type 1 alpha 1 mRNA with an increase in Sox9 mRNA expression especially in the presence of a platelet lysate, suggesting early chondrogenesis.

Conclusion: Fibrin/HA-MA hydrogel may be a suitable delivery method for BMSCs, inducing BMSC differentiation into chondrocytes and potentially aiding in articular cartilage repair for OA therapy.

No MeSH data available.


Related in: MedlinePlus

Compressive modulus of fibrin/HA-MA hydrogels at 20% strain. *Statistically significant difference between successive HA-MA concentrations for a given fibrinogen concentration (p < 0.05). † – Statistically significant difference between different fibrinogen concentrations for a given HA-MA concentration (p < 0.05).
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Figure 5: Compressive modulus of fibrin/HA-MA hydrogels at 20% strain. *Statistically significant difference between successive HA-MA concentrations for a given fibrinogen concentration (p < 0.05). † – Statistically significant difference between different fibrinogen concentrations for a given HA-MA concentration (p < 0.05).

Mentions: To examine mechanical stiffness of the fibrin/HA-MA hydrogels, unconfined compression tests were performed to determine the compressive modulus at 20% strain (Figure 5), as described previously [47]. One-way ANOVA comparison of the data confirmed that the groups were statistically different (p < 0.05). Student’s t-Test was used to find the p value between two groups to determine any statistically significant difference (p < 0.05). Allowing HA-MA concentrations to vary from 0 to 1.5 mg/mL, the compressive modulus of the fibrin/HA-MA hydrogel with 4 mg/mL fibrinogen varied from 1.62 ± 0.6kPa to 4.19 ± 0.28kPa, and the compressive modulus of the fibrin/HA-MA hydrogel with 6 mg/mL fibrinogen varied from 3.39 ± 0.91kPa to 6.76 ± 0.52kPa. Since there was no statistically significant difference between the two conditions 4 mg/mL of fibrinogen with 0 mg/mL of HA-MA and 4 mg/mL of fibrinogen with 0.5 mg/mL of HA-MA (p = 0.1916), we decided to ignore the small apparent difference in the compressive moduli. The 6 mg/mL fibrinogen hydrogel possessed the higher compressive modulus at each HA-MA concentration. Increasing concentrations of both fibrinogen and HA-MA was directly correlated to an increased compressive modulus. Reinforcement with HA-MA crosslinking marginally improved mechanical strength over previously demonstrated pure fibrin hydrogels [42-45]. Since the gel was not stable below 3 mg/mL concentration, we could not measure the compressive strength at the lower concentrations.


A fibrin/hyaluronic acid hydrogel for the delivery of mesenchymal stem cells and potential for articular cartilage repair.

Snyder TN, Madhavan K, Intrator M, Dregalla RC, Park D - J Biol Eng (2014)

Compressive modulus of fibrin/HA-MA hydrogels at 20% strain. *Statistically significant difference between successive HA-MA concentrations for a given fibrinogen concentration (p < 0.05). † – Statistically significant difference between different fibrinogen concentrations for a given HA-MA concentration (p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4109069&req=5

Figure 5: Compressive modulus of fibrin/HA-MA hydrogels at 20% strain. *Statistically significant difference between successive HA-MA concentrations for a given fibrinogen concentration (p < 0.05). † – Statistically significant difference between different fibrinogen concentrations for a given HA-MA concentration (p < 0.05).
Mentions: To examine mechanical stiffness of the fibrin/HA-MA hydrogels, unconfined compression tests were performed to determine the compressive modulus at 20% strain (Figure 5), as described previously [47]. One-way ANOVA comparison of the data confirmed that the groups were statistically different (p < 0.05). Student’s t-Test was used to find the p value between two groups to determine any statistically significant difference (p < 0.05). Allowing HA-MA concentrations to vary from 0 to 1.5 mg/mL, the compressive modulus of the fibrin/HA-MA hydrogel with 4 mg/mL fibrinogen varied from 1.62 ± 0.6kPa to 4.19 ± 0.28kPa, and the compressive modulus of the fibrin/HA-MA hydrogel with 6 mg/mL fibrinogen varied from 3.39 ± 0.91kPa to 6.76 ± 0.52kPa. Since there was no statistically significant difference between the two conditions 4 mg/mL of fibrinogen with 0 mg/mL of HA-MA and 4 mg/mL of fibrinogen with 0.5 mg/mL of HA-MA (p = 0.1916), we decided to ignore the small apparent difference in the compressive moduli. The 6 mg/mL fibrinogen hydrogel possessed the higher compressive modulus at each HA-MA concentration. Increasing concentrations of both fibrinogen and HA-MA was directly correlated to an increased compressive modulus. Reinforcement with HA-MA crosslinking marginally improved mechanical strength over previously demonstrated pure fibrin hydrogels [42-45]. Since the gel was not stable below 3 mg/mL concentration, we could not measure the compressive strength at the lower concentrations.

Bottom Line: This chondrogenic hydrogel system can be delivered in a minimally invasive manner through a small gauge needle, forming a three-dimensional (3D) network structure in situ.However, an ongoing problem with fibrin/HA-based biomaterials is poor mechanical strength.Quantitative polymerase chain reaction (qPCR) of BMSCs incubated in the fibrin/HA-MA hydrogel confirmed decreased expression of collagen type 1 alpha 1 mRNA with an increase in Sox9 mRNA expression especially in the presence of a platelet lysate, suggesting early chondrogenesis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Bioengineering Department, University of Colorado, Anschutz Medical Campus, Mail Stop 8607, 12700 East 19th Avenue, Aurora, CO 80045, USA ; Regenerative Sciences, 403 Summit Blvd, Suite 201, Broomfield, CO 80021, USA.

ABSTRACT

Background: Osteoarthritis (OA) is a degenerative joint disease affecting approximately 27 million Americans, and even more worldwide. OA is characterized by degeneration of subchondral bone and articular cartilage. In this study, a chondrogenic fibrin/hyaluronic acid (HA)-based hydrogel seeded with bone marrow-derived mesenchymal stem cells (BMSCs) was investigated as a method of regenerating these tissues for OA therapy. This chondrogenic hydrogel system can be delivered in a minimally invasive manner through a small gauge needle, forming a three-dimensional (3D) network structure in situ. However, an ongoing problem with fibrin/HA-based biomaterials is poor mechanical strength. This was addressed by modifying HA with methacrylic anhydride (MA) (HA-MA), which reinforces the fibrin gel, thereby improving mechanical properties. In this study, a range of fibrinogen (the fibrin precursor) and HA-MA concentrations were explored to determine optimal conditions for increased mechanical strength, BMSC proliferation, and chondrogenesis potential in vitro.

Results: Increased mechanical strength was achieved by HA-MA reinforcement within fibrin hydrogels, and was directly correlated with increasing HA-MA concentration. Live/dead staining and metabolic assays confirmed that the crosslinked fibrin/HA-MA hydrogels provided a suitable 3D environment for BMSC proliferation. Quantitative polymerase chain reaction (qPCR) of BMSCs incubated in the fibrin/HA-MA hydrogel confirmed decreased expression of collagen type 1 alpha 1 mRNA with an increase in Sox9 mRNA expression especially in the presence of a platelet lysate, suggesting early chondrogenesis.

Conclusion: Fibrin/HA-MA hydrogel may be a suitable delivery method for BMSCs, inducing BMSC differentiation into chondrocytes and potentially aiding in articular cartilage repair for OA therapy.

No MeSH data available.


Related in: MedlinePlus