scholarly journals Immunochemical and Mechanical Characterization of Cartilage Subtypes in Rabbit

2002 ◽  
Vol 50 (8) ◽  
pp. 1049-1058 ◽  
Author(s):  
Andreas Naumann ◽  
James E. Dennis ◽  
Amad Awadallah ◽  
David A. Carrino ◽  
Joseph M. Mansour ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Joint Analysis ◽  
Type I ◽  
Auricular Cartilage ◽  
Collagen Types ◽  
Type Vi Collagen ◽  
Intense Staining ◽  
Nasal Cartilage

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.

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 ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Collagen Type ◽  
Cartilage Regeneration ◽  
Collagen Type I ◽  
Collagen I ◽  
Host Cells ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Type I ◽  
A Cell

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.

scholarly journals Characterization of Collagen Type I and II Blended Hydrogels for Articular Cartilage Tissue Engineering

2016 ◽  
Vol 17 (10) ◽  
pp. 3145-3152 ◽  
Author(s):  
Nelda Vázquez-Portalatı́n ◽  
Claire E. Kilmer ◽  
Alyssa Panitch ◽  
Julie C. Liu
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Collagen Type ◽  
Cartilage Tissue Engineering ◽  
Collagen Type I ◽  
Cartilage Tissue ◽  
Type I

scholarly journals Adipose-Derived Mesenchymal Stem Cell Chondrospheroids Cultured in Hypoxia and a 3D Porous Chitosan/Chitin Nanocrystal Scaffold as a Platform for Cartilage Tissue Engineering

2020 ◽  
Vol 21 (3) ◽  
pp. 1004 ◽  
Author(s):  
Veronica Zubillaga ◽  
Ana Alonso-Varona ◽  
Susana C. M. Fernandes ◽  
Asier M. Salaberria ◽  
Teodoro Palomares
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Collagen Type ◽  
Porous Scaffold ◽  
Cartilage Tissue Engineering ◽  
Cartilage Regeneration ◽  
Cartilage Tissue ◽  
Type I ◽  
Low Oxygen ◽  
Porous Chitosan

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.

scholarly journals Immunohistochemistry of types I and II collagen in undecalcified skeletal tissues.

1983 ◽  
Vol 31 (3) ◽  
pp. 417-425 ◽  
Author(s):  
W A Horton ◽  
C Dwyer ◽  
R Goering ◽  
D C Dean
Keyword(s):  
Articular Cartilage ◽  
Growth Plate ◽  
Subchondral Bone ◽  
Type I Collagen ◽  
Sodium Ethoxide ◽  
Cartilage Matrix ◽  
Antigenic Determinants ◽  
Immunoperoxidase Staining ◽  
Type I ◽  
Intense Staining

Types I and II collagen were demonstrated in semithin sections of undecalcified human endochondral growth plate, articular cartilage, and subchondral bone. The effects of several different methods for fixation, embedding, exposing of antigenic determinants, and immunoperoxidase staining were examined. Fixation in buffered formalin and paraformaldehyde-lysine-periodate solution gave more intense staining for collagens than fixation in paraformaldehyde-gluaraldehyde or Bouin's solution. Specimens embedded in Spurr epoxy resin yielded intense and uniform staining of areas known to contain the particular collagens after the resin had been removed by sodium ethoxide. The staining was enchanced following enzymatic digestion, especially with protease V (Sigma). Staining sensitivity and specificity were comparable with the indirect conjugate and double peroxidase-antiperoxidase (PAP) techniques; the PAP method was less sensitive. Embedment in methacrylate resins proved unsatisfactory because of exaggerated immunostaining of mineralized sites in comparison to unmineralized areas of the same tissues. In the growth plate specimens, type I collagen was identified in the matrices of bone, periosteum, perichondrium, and in the cytoplasm of hypertrophic and degenerative chondrocytes. Type II collagen was found uniformly throughout the cartilage matrix and in spicules of unresorbed cartilage matrix located in subchondral bone. A similar staining pattern was observed for the articular cartilage, except that type I collagen was not detected in chondrocytes.

scholarly journals Arthroscopic utilization of ChondroFiller gel for the treatment of hip articular cartilage defects: a cohort study with 12- to 60-month follow-up

2021 ◽  
Author(s):  
Jacek Mazek ◽  
Maciej Gnatowski ◽  
Antonio Porthos Salas ◽  
John M O’Donnell ◽  
Marcin Domżalski ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Long Term Results ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Type I ◽  
Treatment Technique ◽  
Acetabular Cartilage ◽  
Cartilage Lesions ◽  
Post Surgery

Abstract ChondroFiller gel is an absorbable collagen implant. It serves as a protective cover for the cartilage defects, allowing chondrocyte migration into the lesion. The implant consists of collagen (Type I) and is derived from veterinary monitored rats. This study evaluates the use of ChondroFiller gel in the treatment of cartilage lesions during hip joint arthroscopy. A prospective study was conducted on a group of 26 adult patients. All patients had an existing femoroacetabular impingement together with acetabular cartilage lesions >2 cm2. All patients underwent hip arthroscopic surgery and the lesions were treated using ChondroFiller gel. The cartilage tissue healing was evaluated postoperatively using MRI. A total of 26 patients, including 5 females and 21 males, all with articular cartilage lesions, were included in the study. Cartilage healing conditions were evaluated for all patients, and the difference between pre- and post-surgery conditions was statistically significant. The follow-up scores have been acquired from 21 out of initial 26 patients (2 were disqualified after receiving THR, 3 could not be reached by researchers) after 3, 4 and 5 years consecutively with 17/21 patients having good/excellent results. The use of ChondroFiller gel during arthroscopy of the hip for acetabular cartilage lesions is an effective treatment technique. Encouraging long-term results have been observed, but further research on larger group of patient is required to better assess the full value of this technique. Patients with pre-existing osteoarthritis (Tönnis 2–3) have poor results.

Blocking Wnt/β-Catenin Signaling Alleviates Osteoarthritis in Rats

2020 ◽  
Vol 10 (5) ◽  
pp. 704-708
Author(s):  
Ying Wang ◽  
Lili Sun ◽  
Junli Qin ◽  
Meng Wu ◽  
Jinfeng Zhang
Keyword(s):  
Articular Cartilage ◽  
Signaling Pathway ◽  
Cartilage Damage ◽  
Iodoacetic Acid ◽  
Protective Role ◽  
Joint Disease ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Real Time Quantitative Pcr ◽  
Safranin O

Osteoarthritis is a chronic multiple degenerative bone and joint disease. The most common manifestations are damaged articular cartilage, thickening and sclerosis of subchondral bone, and osteophytes formation at the joint edge. This study will block the Wnt/β -catenin signaling pathway to observe its role on a rat model of osteoarthritis. In this experiment, rat osteoarthritis model was established by intra-articular administration of iodoacetic acid. The LF-3 treatment was started on the second day after modeling. Two weeks later, rat knee articular cartilage was collected. The morphology of cartilage was observed by HE, Safranin O and toluidine blue staining. The expression levels of TIMP-3, MMP-9 protein and mRNA were detected by western blot and real-time quantitative PCR. After blocking the Wnt/ β-catenin signaling pathway, TIMP-3 expression level was increased, MMP-9 expression was decreased, and cartilage damage was ameliorated. Blocking the Wnt/ β-catenin signal can reduce the degradation of matrix in cartilage tissue, thereby playing a protective role and helping the recovery of osteoarthritic rats.

scholarly journals Interstitial flow upregulates matrix synthesis and attenuates NF-kB signaling in a novel perfusion bioreactor for articular cartilage tissue engineering

2021 ◽  
Author(s):  
Haneen Abusharkh ◽  
Terreill Robertson ◽  
Juana Mendenhall ◽  
Bulent Gozen ◽  
Edwin Tingstad ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Cell Viability ◽  
Fold Increase ◽  
Cartilage Tissue ◽  
Type I ◽  
Interstitial Flow ◽  
Perfusion System ◽  
Matrix Synthesis ◽  
Porous Chitosan

The present study is focused on designing an easy-to-use novel perfusion system for articular cartilage (AC) tissue engineering and using it to elucidate the mechanism by which interstitial shear upregulates matrix synthesis by articular chondrocytes (AChs). Porous chitosan-agarose (CHAG) scaffolds were synthesized, freeze-dried, and compared to bulk agarose (AG) scaffolds. Both scaffold types were seeded with osteoarthritic human AChs and cultured in a novel perfusion system for one week with a shear-inducing medium flow velocity of 0.33 mm/s corresponding to an average surficial shear of 0.4 mPa and a CHAG interstitial shear of 40 mPa. While there were no statistical differences in cell viability for perfusion vs. static cultures for either scaffold type, CHAG scaffold cultures exhibited 3.3-fold higher (p<0.005) cell viability compared to AG scaffold cultures. Effects of combined superficial and interstitial perfusion for CHAG showed 150- and 45-fold (p<0.0001) increases in total collagen (COL) and 13- and 2.2-fold (p<0.001) increases in glycosaminoglycans (GAGs) over AG’s scaffold non-perfusion and perfusion cultures, respectively, and a 1.5-fold and 3.6-fold (p<0.005) increase over non-perfusion CHAG cultures. Contrasting CHAG perfusion and static cultures, chondrogenic gene comparisons showed a 3.5-fold increase in collagen type II/type I (COL2A1/COL1A1) mRNA ratio (p<0.05), and a 1.3-fold increase in aggrecan mRNA. Observed effects are suggested to be the result of inhibiting the inflammatory NF-κB signal transduction pathway as confirmed by a further study that indicated a reduction by 3.2-fold (p<0.05) upon exposure to perfusion. Our results demonstrate that the presence of pores plays a critical role in improving cell viability and that interstitial flow caused by medium perfusion through the porous scaffolds enhances the expression of chondrogenic genes and ECM components through the downregulation of NF-κB1.

scholarly journals Main and Minor Types of Collagens in the Articular Cartilage: The Role of Collagens in Repair Tissue Evaluation in Chondral Defects

2021 ◽  
Vol 22 (24) ◽  
pp. 13329
Author(s):  
Lourdes Alcaide-Ruggiero ◽  
Verónica Molina-Hernández ◽  
M. M. Granados ◽  
J. M. Domínguez
Keyword(s):  
Articular Cartilage ◽  
Hyaline Cartilage ◽  
Cartilage Tissue ◽  
Type Ii ◽  
Collagen Types ◽  
Chondral Defects ◽  
Healthy Cartilage ◽  
Hyaline Articular Cartilage

Several collagen subtypes have been identified in hyaline articular cartilage. The main and most abundant collagens are type II, IX and XI collagens. The minor and less abundant collagens are type III, IV, V, VI, X, XII, XIV, XVI, XXII, and XXVII collagens. All these collagens have been found to play a key role in healthy cartilage, regardless of whether they are more or less abundant. Additionally, an exhaustive evaluation of collagen fibrils in a repaired cartilage tissue after a chondral lesion is necessary to determine the quality of the repaired tissue and even whether or not this repaired tissue is considered hyaline cartilage. Therefore, this review aims to describe in depth all the collagen types found in the normal articular cartilage structure, and based on this, establish the parameters that allow one to consider a repaired cartilage tissue as a hyaline cartilage.

scholarly journals Histochemistry of Equine Damaged Tendons, Ligaments and Articular Cartilage

2018 ◽  
Vol 46 (1) ◽  
pp. 8
Author(s):  
Gabriela De Bastiani ◽  
Flávio Desessards De La Corte ◽  
Karin Erica Brass ◽  
Camila Cantarelli ◽  
Stefano Dau ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Connective Tissue ◽  
Alcian Blue ◽  
Collagen Type ◽  
Repair Process ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type Iii Collagen ◽  
Type I ◽  
Loose Connective Tissue

Background: The injury repair process in tendons and ligaments includes different phases such as inflammation, neovascularization, fibroblast proliferation and fibrosis. Collagen type and tissue characteristics of tendon and ligament repair are described such as type collagen differentiation and properties of the scars tissue. The degeneration of articular cartilage when, characterized by loss of the articular layers associated of the decreased of proteoglycans. The aim of this study is to describe by histochemistry techniques the characteristics of tissue scar, collagen type in the repair process of tendons and ligaments, as well as articular cartilage degeneration.Materials, Methods & Results: Tissue samples of equine tendons, ligaments and articular cartilage of the metacarpophalangeal joint region were evaluated by ultrasonography, macroscopically and prepared for routine histopathology (H&E staining). The inclusion criterion of the samples in this study was based on the presence of lesions characterized in H&E stain as fibroplasia, neovascularization, collagenolysis, chondroid metaplasia in tendons and ligaments and fibrillation and cartilaginous eburnation lesions in the articular cartilage samples. The Masson’s trichrome, Picrosirius red and Alcian blue staining techniques were also performed in addition to H&E. Pathologic findings in the tendons and ligaments included fibroplasia, collagenolysis, chondroid metaplasia and lymphohistioplasmacytic inflammation. Tendons and ligaments scars were composed of type III collagen but there was also some type I collagen. Fiber alignment of tendons and ligaments in the reorganization tissue was not flawless and the fiber appearance was characterized by a lack of the fiber crimp and parallelism. The fibroplasia was characterized by endotendinous tickening areas associated with the presence of loose connective tissue. In the areas of loose connective tissue substitution, collagen type fibers are intercalated to a lesser extent by type-III collagen fibers. In the Alcian blue stained samples of articular cartilage observed the surface layer and the matrix zone of calcified cartilage were weakly stained in blue.Discussion: Three special stains were utilized in this study along with the H&E evaluation elucidating the behavior tendons, ligaments and articular cartilage injury. The important observation in this study was fibroplasia in tendons and ligaments seems to be composed by abundant of loose connective tissue, chondrocytes and intermingled collagen type I and III fibers associated with lack of crimps alignment of the fibers. The fragile structure suggested by the Masson’s trichrome stain results (presence of the loose connective tissue) in this study perhaps make the tendons and ligaments receptive to other lesions. The characteristic blue discoloration of collagen fibers was only observed in the loose connective tissue may be because the dye penetration becomes easier when compared to the dense connective tissue (stained in red). The Masson’s trichrome made possible the differentiated the dense connective tissue of the loose connective tissue. The combined histochemistry staining technics allowed an improved characterization of fiber alignment, collagen type, inflammatory cell infiltration and neovascularization, which happens during the repair process of tendons and ligaments. The fibrillation and eburnation of the articular cartilage were associated with the decrease Alcian Blue staining characterized by degeneration process of articular cartilage.

scholarly journals Oral Administration of Salmon Cartilage Proteoglycan Attenuates Osteoarthritis in a Monosodium Iodoacetate-Induced Rat Model

2020 ◽  
Vol 15 (12) ◽  
pp. 1934578X2098211
Author(s):  
Tuyen Danh Le ◽  
Hien Thi Thu Vu ◽  
Iddamalgoda Arunasiri ◽  
Kenichi Ito ◽  
Tadahiro Makise ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Knee Osteoarthritis ◽  
Rat Model ◽  
Interleukin 1Β ◽  
Cartilage Tissue ◽  
Necrosis Factor Alpha ◽  
Nasal Cartilage ◽  
Tnf Α ◽  
Monosodium Iodoacetate

Proteoglycan (PG) is a type of glycoprotein which forms an extracellular matrix with collagen and hyaluronic acid to maintain articular cartilage, synovial membrane, and synovial fluid. This study aimed to evaluate the antiosteoarthritis effects of salmon nasal cartilage-derived PG in alleviating knee osteoarthritis in an osteoarthritis rat model. Knee osteoarthritis was induced in rats by intra-articular injection of monosodium iodoacetate (MIA), 3 mg/knee, to the right knee. Animals were then administered either diclofenac (3 mg/kg body weight [b.w]/day) or proteoglycan F (PGF; 40 mg/kg and 120 mg/kg b.w/day) by oral gavage for 6 consecutive weeks. Knee diameters were measured throughout the experimental period; serum interleukin-1β and tumor necrosis factor-alpha (TNF-α) levels, and histological analysis of the ligament were carried out at the end of the experiment. Salmon cartilage PG considerably alleviated the osteoarthritis symptoms in the model and lowered the serum concentrations of interleukin-1β and TNF-α. Diclofenac 3 mg/kg/day and PGF at doses of 40 mg/kg/day and 120 mg/kg/day also improved articular cartilage structure on further histological studies. This study demonstrated the in vivo effect of salmon cartilage PG in attenuating symptoms in an MIA-induced rat model, including reduction of inflammatory markers and histological improvement of cartilage tissue.

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