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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

Scanning electron micrographs of (A) 6 mg/mL fibrinogen with no HA-MA at 5000×, (B) 6 mg/mL fibrinogen with no HA-MA at 10,000×, (C) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 200x, and (D) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 5000×. Scale bar: 1 μm.
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Figure 6: Scanning electron micrographs of (A) 6 mg/mL fibrinogen with no HA-MA at 5000×, (B) 6 mg/mL fibrinogen with no HA-MA at 10,000×, (C) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 200x, and (D) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 5000×. Scale bar: 1 μm.

Mentions: Fibrinogen (fibrin-precursor) and HA-MA form a gel with an interpenetrating network of fibers through ionic and chemical interactions [34,51,52]. The hydrogel was then reinforced using UV photocrosslinking, which was achieved by photocrosslinking through the polymerizable vinyl group (–C = C–) present on the methacrylate side chain. The fibrin/HA-MA hydrogel even after crosslinking would still be completely biodegradable since the degradation of the hydrogel could be achieved through the hydrolysis of the ester bond present in the methacrylate side chain and the ether group present in the main hyaluronic acid backbone. To gain further insight into the structure of the hydrogels, field emission scanning electron microscopy (FESEM) was performed (Figure 6). The fibrin gels without HA-MA (Figures 6A and B) showed fibrous structure with pore sizes on the order of 1 μm. Addition of HA-MA resulted in a more sheet-like morphology with pore sizes in the range of 10-100 μm (Figure 6C). Magnifying further into the fibrin/HA-MA construct showed that fibrin fibers were still embedded within the structure (Figure 6D), indicating that the gelation occurred as intended by design: formation of fibrin gel followed by HA-MA reinforcement and crosslinking. Since 6 mg/mL fibrinogen with 1 mg/mL HA-MA led to well-controlled proliferation and an increase in mechanical strength, this fibrin/HA-MA hydrogel formulation was used for further study in vitro to determine mRNA expression of key chondrogenic markers.


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)

Scanning electron micrographs of (A) 6 mg/mL fibrinogen with no HA-MA at 5000×, (B) 6 mg/mL fibrinogen with no HA-MA at 10,000×, (C) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 200x, and (D) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 5000×. Scale bar: 1 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 6: Scanning electron micrographs of (A) 6 mg/mL fibrinogen with no HA-MA at 5000×, (B) 6 mg/mL fibrinogen with no HA-MA at 10,000×, (C) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 200x, and (D) 6 mg/mL fibrinogen with 1 mg/mL HA-MA at 5000×. Scale bar: 1 μm.
Mentions: Fibrinogen (fibrin-precursor) and HA-MA form a gel with an interpenetrating network of fibers through ionic and chemical interactions [34,51,52]. The hydrogel was then reinforced using UV photocrosslinking, which was achieved by photocrosslinking through the polymerizable vinyl group (–C = C–) present on the methacrylate side chain. The fibrin/HA-MA hydrogel even after crosslinking would still be completely biodegradable since the degradation of the hydrogel could be achieved through the hydrolysis of the ester bond present in the methacrylate side chain and the ether group present in the main hyaluronic acid backbone. To gain further insight into the structure of the hydrogels, field emission scanning electron microscopy (FESEM) was performed (Figure 6). The fibrin gels without HA-MA (Figures 6A and B) showed fibrous structure with pore sizes on the order of 1 μm. Addition of HA-MA resulted in a more sheet-like morphology with pore sizes in the range of 10-100 μm (Figure 6C). Magnifying further into the fibrin/HA-MA construct showed that fibrin fibers were still embedded within the structure (Figure 6D), indicating that the gelation occurred as intended by design: formation of fibrin gel followed by HA-MA reinforcement and crosslinking. Since 6 mg/mL fibrinogen with 1 mg/mL HA-MA led to well-controlled proliferation and an increase in mechanical strength, this fibrin/HA-MA hydrogel formulation was used for further study in vitro to determine mRNA expression of key chondrogenic markers.

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