Roles of Silk Fibroin on Characteristics of Hyaluronic Acid/Silk Fibroin Hydrogels for Tissue Engineering of Nucleus Pulposus

Silk fibroin (SF) and hyaluronic acid (HA) were crosslinked by horseradish peroxidase (HRP)/H2O2, and 1,4-Butanediol di-glycidyl ether (BDDE), respectively, to produce HA/SF-IPN (interpenetration network) (HS-IPN) hydrogels. HS-IPN hydrogels consisted of a SF strain with a high content of tyrosine (e.g., strain A) increased viscoelastic modules compared with those with low contents (e.g., strain B and C). Increasing the quantities of SF in HS-IPN hydrogels (e.g., HS7-IPN hydrogels with weight ratio of HA/SF, 5:7) increased viscoelastic modules of the hydrogels. In addition, the mean pores size of scaffolds of the model hydrogels were around 38.96 ± 5.05 μm which was between those of scaffolds H and S hydrogels. Since the viscoelastic modulus of the HS7-IPN hydrogel were similar to those of human nucleus pulposus (NP), it was chosen as the model hydrogel for examining the differentiation of human bone marrow-derived mesenchymal stem cell (hBMSC) to NP. The differentiation of hBMSC induced by transforming growth factor β3 (TGF-β3) in the model hydrogels to NP cells for 7 d significantly enhanced the expressions of glycosaminoglycan (GAG) and collagen type II, and gene expressions of aggrecan and collagen type II while decreased collagen type I compared with those in cultural wells. In summary, the model hydrogels consisted of SF of strain A, and high concentrations of SF showed the highest viscoelastic modulus than those of others produced in this study, and the model hydrogels promoted the differentiation of hBMSC to NP cells.

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Limb bud mesenchyme cultured under tensile strain remodel collagen type I tubes to produce fibrillar collagen type II

Biorheology ◽

10.3233/bir-2009-0553 ◽

2009 ◽

Vol 46 (6) ◽

pp. 439-450 ◽

Cited By ~ 1

Author(s):

Jennifer R. Amos ◽

Shigeng Li ◽

Michael Yost ◽

Harry Phloen ◽

Jay D. Potts

Keyword(s):

Tensile Strain ◽

Collagen Type ◽

Collagen Type I ◽

Limb Bud ◽

Type I ◽

Fibrillar Collagen ◽

Type Ii ◽

Collagen Type Ii

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Maintenance of chondrocyte phenotype during expansion on PLLA microtopographies

Journal of Tissue Engineering ◽

10.1177/2041731418789829 ◽

2018 ◽

Vol 9 ◽

pp. 204173141878982 ◽

Cited By ~ 6

Author(s):

Elisa Costa ◽

Cristina González-García ◽

José Luis Gómez Ribelles ◽

Manuel Salmerón-Sánchez

Keyword(s):

Lactic Acid ◽

Type I Collagen ◽

Collagen Type ◽

Articular Chondrocytes ◽

Collagen Type I ◽

Cell Phenotype ◽

Type I ◽

Type Ii ◽

Collagen Type Ii ◽

Chondrocyte Phenotype

Articular chondrocytes are difficult to grow, as they lose their characteristic phenotype following expansion on standard tissue culture plates. Here, we show that culturing them on surfaces of poly(L-lactic acid) of well-defined microtopography allows expansion and maintenance of characteristic chondrogenic markers. We investigated the dynamics of human chondrocyte dedifferentiation on the different poly(L-lactic acid) microtopographies by the expression of collagen type I, collagen type II and aggrecan at different culture times. When seeded on poly(L-lactic acid), chondrocytes maintained their characteristic hyaline phenotype up to 7 days, which allowed to expand the initial cell population approximately six times without cell dedifferentiation. Maintenance of cell phenotype was afterwards correlated to cell adhesion on the different substrates. Chondrocytes adhesion occurs via the α5 β1 integrin on poly(L-lactic acid), suggesting cell–fibronectin interactions. However, α2 β1 integrin is mainly expressed on the control substrate after 1 day of culture, and the characteristic chondrocytic markers are lost (collagen type II expression is overcome by the synthesis of collagen type I). Expanding chondrocytes on poly(L-lactic acid) might be an effective solution to prevent dedifferentiation and improving the number of cells needed for autologous chondrocyte transplantation.

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Limb bud mesenchyme cultured under tensile strain remodel collagen type I tubes to produce fibrillar collagen type II

Biorheology ◽

10.3233/bir-2010-0571 ◽

2010 ◽

Vol 47 (2) ◽

pp. 163-163

Author(s):

Jennifer R. Amos ◽

Shigeng Li ◽

Michael Yost ◽

Harry Phloen ◽

Jay D. Potts

Keyword(s):

Tensile Strain ◽

Collagen Type ◽

Collagen Type I ◽

Limb Bud ◽

Type I ◽

Fibrillar Collagen ◽

Type Ii ◽

Collagen Type Ii

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Enhanced Early Tissue Regeneration after Matrix-Assisted Autologous Mesenchymal Stem Cell Transplantation in Full Thickness Chondral Defects in a Minipig Model

Cell Transplantation ◽

10.3727/096368909x471297 ◽

2009 ◽

Vol 18 (8) ◽

pp. 923-932 ◽

Cited By ~ 59

Author(s):

Martin Jung ◽

Balazs Kaszap ◽

Anna Redöhl ◽

Eric Steck ◽

Steffen Breusch ◽

...

Keyword(s):

Articular Cartilage ◽

Collagen Type ◽

Type I ◽

Type Ii ◽

Collagen Type Ii ◽

Histomorphometric Analysis ◽

Tissue Quality ◽

Repair Tissue ◽

Adult Mesenchymal Stem Cells ◽

Defect Area

Adult mesenchymal stem cells (MSCs) are an attractive cell source for new treatment strategies in regenerative medicine. This study investigated the potential effect of matrix assisted MSC transplantation for articular cartilage regeneration in a large-animal model 8 weeks postoperatively. MSCs from bone marrow aspirates of eight Goettingen minipigs were isolated and expanded prior to surgery. Articular cartilage defects of 5.4 mm were created bilaterally in the medial patellar groove without penetrating the subchondral bone plate. Defects were either left empty ( n = 4), covered with a collagen type I/III membrane ( n = 6) or additionally treated with autologous MSC transplantation (2 × 106; n = 6). After 8 weeks animals were euthanized and the defect area was assessed for its gross appearance. Histomorphological analysis of the repair tissue included semiquantitative scoring (O'Driscoll score) and quantitative histomorphometric analysis for its glycosaminoglycan (GAG) and collagen type II content. All membranes were found to cover the defect area 8 weeks postoperatively. Median defect filling was 115.8% (membrane), 117.8% (empty), and 100.4% (MSC), respectively (not significant). Histomorphological scoring revealed significantly higher values in MSC-treated defects (median 16.5) when compared to membrane treatment (median 9.5) or empty defects (median 11.5; p = 0.015 and p = 0.038). Histomorphometric analysis showed larger GAG/collagen type II-positive areas in the MSC-treated group (median 24.6%/29.5% of regeneration tissue) compared to 13.6%/33.1% (empty defects) and 1.7%/6.2% (membrane group; p = 0.066). Cell distribution was more homogeneous in MSC compared to membrane-only group, where cells were found mainly near the subchondral zone. In conclusion, autologous matrix-assisted MSC transplantation significantly increased the histomorphological repair tissue quality during early articular cartilage defect repair and resulted in higher GAG/collagen type II-positive cross-sectional areas of the regenerated tissue.

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