scholarly journals Evaluation of a Cell-Free Collagen Type I-Based Scaffold for Articular Cartilage Regeneration in an Orthotopic Rat Model

Materials ◽  
2020 ◽  
Vol 13 (10) ◽  
pp. 2369 ◽  
Author(s):  
Marta Anna Szychlinska ◽  
Giovanna Calabrese ◽  
Silvia Ravalli ◽  
Anna Dolcimascolo ◽  
Paola Castrogiovanni ◽  
...  

The management of chondral defects represents a big challenge because of the limited self-healing capacity of cartilage. Many approaches in this field obtained partial satisfactory results. Cartilage tissue engineering, combining innovative scaffolds and stem cells from different sources, emerges as a promising strategy for cartilage regeneration. The aim of this study was to evaluate the capability of a cell-free collagen I-based scaffold to promote cartilaginous repair after orthotopic implantation in vivo. Articular cartilage lesions (ACL) were created at the femoropatellar groove in rat knees and cell free collagen I-based scaffolds (S) were then implanted into right knee defect for the ACL-S group. No scaffold was implanted for the ACL group. At 4-, 8- and 16-weeks post-transplantation, degrees of cartilage repair were evaluated by morphological, histochemical and gene expression analyses. Histological analysis shows the formation of fibrous tissue, at 4-weeks replaced by a tissue resembling the calcified one at 16-weeks in the ACL group. In the ACL-S group, progressive replacement of the scaffold with the newly formed cartilage-like tissue is shown, as confirmed by Alcian Blue staining. Immunohistochemical and quantitative real-time PCR (qRT-PCR) analyses display the expression of typical cartilage markers, such as collagen type I and II (ColI and ColII), Aggrecan and Sox9. The results of this study display that the collagen I-based scaffold is highly biocompatible and able to recruit host cells from the surrounding joint tissues to promote cartilaginous repair of articular defects, suggesting its use as a potential approach for cartilage tissue regeneration.

2016 ◽  
Vol 17 (10) ◽  
pp. 3145-3152 ◽  
Author(s):  
Nelda Vázquez-Portalatı́n ◽  
Claire E. Kilmer ◽  
Alyssa Panitch ◽  
Julie C. Liu

2020 ◽  
Vol 21 (3) ◽  
pp. 1004 ◽  
Author(s):  
Veronica Zubillaga ◽  
Ana Alonso-Varona ◽  
Susana C. M. Fernandes ◽  
Asier M. Salaberria ◽  
Teodoro Palomares

Articular cartilage degeneration is one of the most common causes of pain and disability in middle-aged and older people. Tissue engineering (TE) has shown great therapeutic promise for this condition. The design of cartilage regeneration constructs must take into account the specific characteristics of the cartilaginous matrix, as well as the avascular nature of cartilage and its cells’ peculiar arrangement in isogenic groups. Keeping these factors in mind, we have designed a 3D porous scaffold based on genipin-crosslinked chitosan/chitin nanocrystals for spheroid chondral differentiation of human adipose tissue-derived mesenchymal stem cells (hASCs) induced in hypoxic conditions. First, we demonstrated that, under low oxygen conditions, the chondrospheroids obtained express cartilage-specific markers including collagen type II (COL2A1) and aggrecan, lacking expression of osteogenic differentiation marker collagen type I (COL1A2). These results were associated with an increased expression of hypoxia-inducible factor 1α, which positively directs COL2A1 and aggrecan expression. Finally, we determined the most suitable chondrogenic differentiation pattern when hASC spheroids were seeded in the 3D porous scaffold under hypoxia and obtained a chondral extracellular matrix with a high sulphated glycosaminoglycan content, which is characteristic of articular cartilage. These findings highlight the potential use of such templates in cartilage tissue engineering.


2012 ◽  
Vol 24 (03) ◽  
pp. 185-195 ◽  
Author(s):  
Ji Hye Hwang ◽  
On You Kim ◽  
A Ram Kim ◽  
Ji Yeon Bae ◽  
Su Mi Jeong ◽  
...  

Adult articular cartilage tissue has poor capability of self-repair. Therefore, a variety of tissue engineering approaches are motivated by the clinical need for articular repair. Alginate has been used as a biomaterial for cartilage regeneration. The alginate is a natural polymer that is extracted from seaweeds and purification. However, the main drawback is the immune rejection in vivo. To overcome this problem, we have developed the biocompability of alginate using modified Korbutt method. After alginate was purified, purified alginate microcapsules were used in cartilage regeneration. Chondrocytes were seeded in purified and nonpurified alginate microcapsules, and then cell viability, proliferation and phenotype were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to confirm mRNA expression on collagen type I and collagen type II for chondrocytes phenotype. Hematoxylin and eosin (H&E) and Safranin-O histological staining showed tissue growth at the interface during the first 10 days. In this study, chondrocytes in purified alginate microcapsules had higher cell viability, proliferation and more phenotype expression than those in nonpurified alginate microcapsules. The results suggest that the purified alginate microcapsule is useful for cartilage regeneration.


2002 ◽  
Vol 50 (8) ◽  
pp. 1049-1058 ◽  
Author(s):  
Andreas Naumann ◽  
James E. Dennis ◽  
Amad Awadallah ◽  
David A. Carrino ◽  
Joseph M. Mansour ◽  
...  

Cartilage is categorized into three general subgroups, hyaline, elastic, and fibrocartilage, based primarily on morphologic criteria and secondarily on collagen (Types I and II) and elastin content. To more precisely define the different cartilage subtypes, rabbit cartilage isolated from joint, nose, auricle, epiglottis, and meniscus was characterized by immunohistochemical (IHC) localization of elastin and of collagen Types I, II, V, VI, and X, by biochemical analysis of total glycosaminoglycan (GAG) content, and by biomechanical indentation assay. Toluidine blue staining and safranin-O staining were used for morphological assessment of the cartilage subtypes. IHC staining of the cartilage samples showed a characteristic pattern of staining for the collagen antibodies that varied in both location and intensity. Auricular cartilage is discriminated from other subtypes by interterritorial elastin staining and no staining for Type VI collagen. Epiglottal cartilage is characterized by positive elastin staining and intense staining for Type VI collagen. The unique pattern for nasal cartilage is intense staining for Type V collagen and collagen X, whereas articular cartilage is negative for elastin (interterritorially) and only weakly positive for collagen Types V and VI. Meniscal cartilage shows the greatest intensity of staining for Type I collagen, weak staining for collagens V and VI, and no staining with antibody to collagen Type X. Matching cartilage samples were categorized by total GAG content, which showed increasing total GAG content from elastic cartilage (auricle, epiglottis) to fibrocartilage (meniscus) to hyaline cartilage (nose, knee joint). Analysis of aggregate modulus showed nasal and auricular cartilage to have the greatest stiffness, epiglottal and meniscal tissue the lowest, and articular cartilage intermediate. This study illustrates the differences and identifies unique characteristics of the different cartilage subtypes in rabbits. The results provide a baseline of data for generating and evaluating engineered repair cartilage tissue synthesized in vitro or for post-implantation analysis.


2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Rodrigo Mardones ◽  
Alessio Giai Via ◽  
Gennaro Pipino ◽  
Claudio M. Jofre ◽  
Sara Muñoz ◽  
...  

Abstract Background Full-thickness articular cartilage injury of the knee is a major cause of disability. The aim of this study is to assess the outcome of patients treated with differentiated to chondrocytes bone marrow mesenchymal stem cells (BM-MSCs) cultured on a collagen type I/III (Chondro-Gide®) scaffold. The secondary aim was to confirm the absence of adverse events. Methods Fifteen patients (19 knees) with symptomatic full-thickness cartilage defects of the knee were enrolled. Bone marrow was harvested from the iliac crest, BM-MSCs were prepared, and expanded cells were grown in a standard medium or in a standard culture medium containing TGF-β. BM-MSCs differentiated to chondrocytes were seeded in a porcine collagen type I/III scaffold (Chondro-Gide®) and cultured in TGF-β containing media. After 4 weeks, the membrane was sutured on the cartilage defect. All patients underwent plain radiographs (antero-posterior, lateral, and axial view of the patella) and MRI of the affected knee. The Oxford knee score, the Lyhsolm scale, and the VAS score were administered to all patients. At final follow-up a MRI for the study of articular cartilage was undertaken. Results The mean size of the cartilage lesions was 20 × 17 mm (range, 15 × 10 mm–30 × 30 mm). At final follow-up, the median Oxford knee score and Lyhsolm scale scores significantly improved from 29 (range 12–39; SD 7.39) to 45 (range 24–48; SD 5.6) and from 55.5 (range 25–81; SD 17.7) to 94.5 (58–100; SD 10.8), respectively. Pain, according to the VAS score, significantly improved. Sixty percent of patients reported their satisfaction as excellent, 20% as good, 14% as fair, and 1 patient as poor. Conclusion The treatment of full-thickness chondral injuries of the knee with differentiated to chondrocytes BM-MSCs and Chondro-Gide® scaffold showed encouraging outcomes. Further studies involving more patients, and with longer follow-up, are required to evaluate the effectiveness of the treatment and the long-term results.


2010 ◽  
Vol 34 (1) ◽  
pp. 79-83 ◽  
Author(s):  
Karsten Gavenis ◽  
Bernhard Schmidt-Rohlfing ◽  
Stefan Andereya ◽  
Torsten Mumme ◽  
Ulrich Schneider ◽  
...  

2020 ◽  
Vol 133 (23) ◽  
pp. jcs248278 ◽  
Author(s):  
Ana M. Diaz-Espinosa ◽  
Patrick A. Link ◽  
Delphine Sicard ◽  
Ignasi Jorba ◽  
Daniel J. Tschumperlin ◽  
...  

ABSTRACTMatrix resorption is essential to the clearance of the extracellular matrix (ECM) after normal wound healing. A disruption in these processes constitutes a main component of fibrotic diseases, characterized by excess deposition and diminished clearance of fibrillar ECM proteins, such as collagen type I. The mechanisms and stimuli regulating ECM resorption in the lung remain poorly understood. Recently, agonism of dopamine receptor D1 (DRD1), which is predominantly expressed on fibroblasts in the lung, has been shown to accelerate tissue repair and clearance of ECM following bleomycin injury in mice. Therefore, we investigated whether DRD1 receptor signaling promotes the degradation of collagen type I by lung fibroblasts. For cultured fibroblasts, we found that DRD1 agonism enhances extracellular cleavage, internalization and lysosomal degradation of collagen I mediated by cathepsin K, which results in reduced stiffness of cell-derived matrices, as measured by atomic force microscopy. In vivo agonism of DRD1 similarly enhanced fibrillar collagen degradation by fibroblasts, as assessed by tissue labeling with a collagen-hybridizing peptide. Together, these results implicate DRD1 agonism in fibroblast-mediated collagen clearance, suggesting an important role for this mechanism in fibrosis resolution.This article has an associated First Person interview with the first author of the paper.


2011 ◽  
Vol 300 (4) ◽  
pp. C907-C918 ◽  
Author(s):  
Matilde Alique ◽  
Laura Calleros ◽  
Alicia Luengo ◽  
Mercedes Griera ◽  
Miguel Ángel Iñiguez ◽  
...  

Glomerular diseases are characterized by a sustained synthesis and accumulation of abnormal extracellular matrix proteins, such as collagen type I. The extracellular matrix transmits information to cells through interactions with membrane components, which directly activate many intracellular signaling events. Moreover, accumulating evidence suggests that eicosanoids derived from cyclooxygenase (COX)-2 participate in a number of pathological processes in immune-mediated renal diseases, and it is known that protein kinase B (AKT) may act through different transcription factors in the regulation of the COX-2 promoter. The present results show that progressive accumulation of collagen I in the extracellular medium induces a significant increase of COX-2 expression in human mesangial cells, resulting in an enhancement in PGE2 production. COX-2 overexpression is due to increased COX-2 mRNA levels. The study of the mechanism implicated in COX-2 upregulation by collagen I showed focal adhesion kinase (FAK) activation. Furthermore, we observed that the activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway by collagen I and collagen I-induced COX-2 overexpression was abolished by PI3K and AKT inhibitors. Additionally, we showed that the cAMP response element (CRE) transcription factor is implicated. Finally, we studied COX-2 expression in an animal model, NG-nitro-l-arginine methyl ester hypertensive rats. In renal tissue and vascular walls, COX-2 and collagen type I content were upregulated. In summary, our results provide evidence that collagen type I increases COX-2 expression via the FAK/PI3K/AKT/cAMP response element binding protein signaling pathway.


2020 ◽  
Author(s):  
Rodrigo Mardones ◽  
Alessio Giai Via ◽  
Gennaro Pipino ◽  
Claudio Jofrè ◽  
Sara Muñoz ◽  
...  

Abstract Background Full-thickness articular cartilage injury of the knee is a major cause of disability. The aim of this study is to assess the results of patients treated with differentiated to chondrocytes bone marrow Mesenchymal Stem Cells (BM-MSCs) cultured on a collagen Type I/III (Chondro-Gide®) scaffold. The secondary aim was to confirm the absence of adverse events. Methods Fifteen patients (19 knees) with symptomatic full-thickness cartilage defects of the knee have been enrolled for the study. Bone marrow was harvested from the iliac crest, BM-MSCs were prepared, and expanded cells were grown in a standard medium or in a standard culture medium containing TGF-β. BM-MSCs differentiated to chondrocytes were seeded in a porcine collagen Type I/III scaffold (Chondro-Gide®), and cultured in TGF- β containing media. After 4 weeks, the membrane was sutured on the cartilage defect. All patients underwent plain radiographs of the knee (antero-posterior, lateral, and axial view of the patella), and MRI of the affected knee. The Oxford knee score, the Lyhsolm scale, and the VAS score were administered to all patients. At final follow-up a MRI for the study of articular cartilage was undertaken. Results The mean size of the cartilage lesions was 20 × 17 mm (range, 15 × 10 mm − 30 × 30 mm). At final follow-up, the median Oxford knee score and Lyhsolm scale scores significantly improved from 29 (range 12–39; SD 7,39) to 45 (range 24–48; SD 5,6) and from 55.5 (range 25–81; SD 17,7) to 94.5 (58–100; SD 10,8) respectively. Pain, according the VAS score, significantly improved. 60% of patients reported their satisfaction as excellent, 20% as good, 14% as fair, and 1 patient as poor. Conclusion The treatment of full-thickness chondral injuries of the knee with differentiated to chondrocytes BM-MSCs and Chondro-Gide® scaffold showed encouraging outcomes. Further studies involving more patients, and with longer follow-up, are required in order to evaluate the effectiveness of the treatment and the long-term results.


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