Articular cartilage repair with recombinant human type II collagen/polylactide scaffold in a preliminary porcine study

Background: Insufficient cell numbers still present a challenge for articular cartilage repair. Converting heterotopic auricular chondrocytes by extracellular matrix may be the solution. Hypothesis: Specific extracellular matrix may convert the phenotype of auricular chondrocytes toward articular cartilage for repair. Study Design: Controlled laboratory study. Methods: For in vitro study, rabbit auricular chondrocytes were cultured in monolayer for several passages until reaching status of dedifferentiation. Later, they were transferred to chondrogenic type II collagen (Col II)–coated plates for further cell conversion. Articular chondrogenic profiles, such as glycosaminoglycan deposition, articular chondrogenic gene, and protein expression, were evaluated after 14-day cultivation. Furthermore, 3-dimensional constructs were fabricated using Col II hydrogel-associated auricular chondrocytes, and their histological and biomechanical properties were analyzed. For in vivo study, focal osteochondral defects were created in the rabbit knee joints, and auricular Col II constructs were implanted for repair. Results: The auricular chondrocytes converted by a 2-step protocol expressed specific profiles of chondrogenic molecules associated with articular chondrocytes. The histological and biomechanical features of converted auricular chondrocytes became similar to those of articular chondrocytes when cultivated with Col II 3-dimensional scaffolds. In an in vivo animal model of osteochondral defects, the treated group (auricular Col II) showed better cartilage repair than did the control groups (sham, auricular cells, and Col II). Histological analyses revealed that cartilage repair was achieved in the treated groups with abundant type II collagen and glycosaminoglycans syntheses rather than elastin expression. Conclusion: The study confirmed the feasibility of applying heterotopic chondrocytes for cartilage repair via extracellular matrix–induced cell conversion. Clinical Relevance: This study proposes a feasible methodology to convert heterotopic auricular chondrocytes for articular cartilage repair, which may serve as potential alternative sources for cartilage repair.

Download Full-text

Natural Type II Collagen Hydrogel, Fibrin Sealant, and Adipose-Derived Stem Cells as a Promising Combination for Articular Cartilage Repair

Cartilage ◽

10.1177/1947603516675914 ◽

2016 ◽

Vol 8 (4) ◽

pp. 439-443 ◽

Cited By ~ 8

Author(s):

Mariana Lazarini ◽

Pedro Bordeaux-Rego ◽

Renata Giardini-Rosa ◽

Adriana S. S. Duarte ◽

Mariana Ozello Baratti ◽

...

Keyword(s):

Stem Cells ◽

Articular Cartilage ◽

Cartilage Repair ◽

Fibrin Sealant ◽

Type Ii Collagen ◽

Adipose Derived Stem Cells ◽

Type Ii ◽

Articular Cartilage Repair ◽

Collagen Hydrogel

Objective Articular cartilage is an avascular tissue with limited ability of self-regeneration and the current clinical treatments have restricted capacity to restore damages induced by trauma or diseases. Therefore, new techniques are being tested for cartilage repair, using scaffolds and/or stem cells. Although type II collagen hydrogel, fibrin sealant, and adipose-derived stem cells (ASCs) represent suitable alternatives for cartilage formation, their combination has not yet been investigated in vivo for focal articular cartilage defects. We performed a simple experimental procedure using the combination of these 3 compounds on cartilage lesions of rabbit knees. Design The hydrogel was developed in house and was first tested in vitro for chondrogenic differentiation. Next, implants were performed in chondral defects with or without ASCs and the degree of regeneration was macroscopically and microscopically evaluated. Results Production of proteoglycans and the increased expression of collagen type II (COL2α1), aggrecan (ACAN), and sex-determining region Y-box 9 (SOX9) confirmed the chondrogenic character of ASCs in the hydrogel in vitro. Importantly, the addition of ASC induced a higher overall repair of the chondral lesions and a better cellular organization and collagen fiber alignment compared with the same treatment without ASCs. This regenerating tissue also presented the expression of cartilage glycosaminoglycan and type II collagen. Conclusions Our results indicate that the combination of the 3 compounds is effective for articular cartilage repair and may be of future clinical interest.

Download Full-text

Comparative Effectiveness of Structural versus Regulatory Protein Gene Transfer on Articular Chondrocyte Matrix Gene Expression

Cartilage ◽

10.1177/1947603517719317 ◽

2017 ◽

Vol 10 (1) ◽

pp. 102-110 ◽

Cited By ~ 1

Author(s):

Shuiliang Shi ◽

Congrong Wang ◽

Albert Chan ◽

Kashif Kirmani ◽

George J. Eckert ◽

...

Keyword(s):

Articular Cartilage ◽

Gene Transfer ◽

Cartilage Repair ◽

Articular Chondrocytes ◽

Type Ii Collagen ◽

Matrix Proteins ◽

Type Ii ◽

Articular Cartilage Repair ◽

Regulatory Factors ◽

Igf I

Objective The production of extracellular matrix is a necessary component of articular cartilage repair. Gene transfer is a promising method to improve matrix biosynthesis by articular chondrocytes. Gene transfer may employ transgenes encoding regulatory factors that stimulate the production of matrix proteins, or may employ transgenes that encode the proteins themselves. The objective of this study was to determine which of these 2 approaches would be the better choice for further development. We compared these 2 approaches using the transgenes encoding the structural matrix proteins, aggrecan or type II collagen, and the transgene encoding the anabolic factor, insulin-like growth factor I (IGF-I). Methods We transfected adult bovine articular chondrocytes with constructs encoding type II collagen, aggrecan, or IGF-I, and measured the expression of type II collagen ( COL2A1) and aggrecan ( ACAN) from their native genes and from their transgenes. Results IGF-I gene ( IGF1) transfer increased the expression of the native chondrocyte COL2A1 and ACAN genes 2.4 and 2.9 times control, respectively. COL2A1 gene transfer did not significantly increase COL2A1 transcripts, even when the transgene included the genomic COL2A1 regulatory sequences stimulated by chondrogenic growth factors. In contrast, ACAN gene transfer increased ACAN transcripts up to 3.4 times control levels. IGF1, but not ACAN, gene transfer increased aggrecan protein production. Conclusion Taken together, these results suggest that the type II collagen and aggrecan production required for articular cartilage repair will be more effectively achieved by genes that encode anabolic regulatory factors than by genes that encode the matrix molecules themselves.

Download Full-text

Cross-Validation of Cyanogen Bromide-Peptide Ratios to Measure the Proportion of Type II Collagen in Pepsin Digests of Equine Articular Cartilage, Meniscus, and Cartilage Repair Tissue

Analytical Biochemistry ◽

10.1006/abio.1994.1025 ◽

1994 ◽

Vol 216 (1) ◽

pp. 195-204 ◽

Cited By ~ 6

Author(s):

R.J. Todhunter ◽

J.A.M. Wootton ◽

N. Altman ◽

G. Lust ◽

R.R. Minor

Keyword(s):

Articular Cartilage ◽

Cartilage Repair ◽

Cyanogen Bromide ◽

Cross Validation ◽

Type Ii Collagen ◽

Type Ii ◽

Repair Tissue ◽

And Cartilage

Download Full-text

PVA‐Based Hydrogels: Promising Candidates for Articular Cartilage Repair

Macromolecular Bioscience ◽

10.1002/mabi.202100147 ◽

2021 ◽

pp. 2100147

Author(s):

Yuru Chen ◽

Jian Song ◽

Song Wang ◽

Weiqiang Liu

Keyword(s):

Articular Cartilage ◽

Cartilage Repair ◽

Articular Cartilage Repair

Download Full-text

Boosting the Intra-Articular Efficacy of Low Dose Corticosteroid through a Biopolymeric Matrix: An In Vivo Model of Osteoarthritis

Cells ◽

10.3390/cells9071571 ◽

2020 ◽

Vol 9 (7) ◽

pp. 1571

Author(s):

Matilde Tschon ◽

Francesca Salamanna ◽

Lucia Martini ◽

Gianluca Giavaresi ◽

Luca Lorenzini ◽

...

Keyword(s):

Articular Cartilage ◽

Pain Sensitivity ◽

Type Ii Collagen ◽

Male Rats ◽

In Vivo Model ◽

Type Ii ◽

Chemical Induction ◽

Local Pain ◽

Gait Parameters

The purpose of this study was to verify the efficacy of a single intra-articular (i.a.) injection of a hyaluronic acid-chitlac (HY-CTL) enriched with two low dosages of triamcinolone acetonide (TA, 2.0 mg/mL and 4.5 mg/mL), in comparison with HY-CTL alone, with a clinical control (TA 40 mg/mL) and with saline solution (NaCl) in an in vivo osteoarthritis (OA) model. Seven days after chemical induction of OA, 80 Sprague Dawley male rats were grouped into five arms (n = 16) and received a single i.a. injection of: 40 mg/mL TA, HY-CTL alone, HY-CTL with 2.0 mg/mL TA (RV2), HY-CTL with 4.5 mg/mL TA (RV4.5) and 0.9% NaCl. Pain sensitivity and Catwalk were performed at baseline and at 7, 14 and 21 days after the i.a. treatments. The histopathology of the joint, meniscus and synovial reaction, type II collagen expression and aggrecan expression were assessed 21 days after treatments. RV4.5 improved the local pain sensitivity in comparison with TA and NaCl. RV4.5 and TA exerted similar beneficial effects in all gait parameters. Histopathological analyses, measured by Osteoarthritis Research Society International (OARSI) and Kumar scores and by immunohistochemistry, evidenced that RV4.5 and TA reduced OA features in the same manner and showed a stronger type II collagen and aggrecan expression; both treatments reduced synovitis, as measured by Krenn score and, at the meniscus level, RV4.5 improved degenerative signs as evaluated by Pauli score. TA or RV4.5 treatments limited the local articular cartilage deterioration in knee OA with an improvement of the physical structure of articular cartilage, gait parameters, the sensitivity to local pain and a reduction of the synovial inflammation.

Download Full-text

Characterization of human type II procollagen and collagen-specific antibodies and their application to the study of human type II collagen processing and ultrastructure

Matrix Biology ◽

10.1016/s0945-053x(97)90114-1 ◽

1997 ◽

Vol 16 (1) ◽

pp. 29-39 ◽

Cited By ~ 7

Author(s):

Sergio A. Jimenez ◽

Leena Ala-Kokko ◽

Darwin J. Prockop ◽

Carmen F. Merryman ◽

Nora Shepard ◽

...

Keyword(s):

Type Ii Collagen ◽

Type Ii ◽

Specific Antibodies ◽

Human Type

Download Full-text

Finite Element Modeling of Osmotic Swelling in Tissue-Engineered Cartilage

ASME 2008 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2008-192368 ◽

2008 ◽

Author(s):

Liming Bian ◽

Terri Ann N. Kelly ◽

Eric G. Lima ◽

Gerard A. Ateshian ◽

Clark T. Hung

Keyword(s):

Articular Cartilage ◽

Potential Role ◽

Swelling Pressure ◽

Type Ii Collagen ◽

Type Ii ◽

Osmotic Swelling ◽

Fixed Charges ◽

Central Regions ◽

Significant Research ◽

Engineered Cartilage

Proteoglycans and Type II collagen represent the two major biochemical constituents of articular cartilage. Collagen fibrils in cartilage resist the swelling pressure that arises from the fixed charges of the glycosaminoglycans (GAGs), and together they give rise to the tissue’s unique load bearing properties. As articular cartilage exhibits a poor intrinsic healing capacity, there is significant research in the development of cell-based therapies for cartilage repair. In some of our tissue engineering studies, we have observed a phenomenon where chondrocyte-seeded hydrogel constructs display cracking in their central regions after significant GAG content has been elaborated in culture. A theoretical analysis was performed to gain greater insights into the potential role that the spatial distribution of proteoglycan and collagen may play in this observed response.

Download Full-text