scholarly journals Progenitor Cells Activated by Platelet Lysate in Human Articular Cartilage as a Tool for Future Cartilage Engineering and Reparative Strategies

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 1052 ◽  
Author(s):  
Simonetta Carluccio ◽  
Daniela Martinelli ◽  
Maria Elisabetta Federica Palamà ◽  
Rui Cruz Pereira ◽  
Roberto Benelli ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Progenitor Cells ◽  
Articular Chondrocytes ◽  
Ex Vivo ◽  
Primary Cultures ◽  
Platelet Lysate ◽  
Cartilage Explants ◽  
Cartilage Tissue ◽  
Human Articular Cartilage

Regenerative strategies for human articular cartilage are still challenging despite the presence of resident progenitor cell population. Today, many efforts in the field of regenerative medicine focus on the use of platelet derivatives due to their ability to reactivate endogenous mechanisms supporting tissue repair. While their use in orthopedics continues, mechanisms of action and efficacy need further characterization. We describe that the platelet lysate (PL) is able to activate chondro-progenitor cells in a terminally differentiated cartilage tissue. Primary cultures of human articular chondrocytes (ACs) and cartilage explants were set up from donor hip joint biopsies and were treated in vitro with PL. PL recruited a chondro-progenitors (CPCs)-enriched population from ex vivo cartilage culture, that showed high proliferation rate, clonogenicity and nestin expression. CPCs were positive for in vitro tri-lineage differentiation and formed hyaline cartilage-like tissue in vivo without hypertrophic fate. Moreover, the secretory profile of CPCs was analyzed, together with their migratory capabilities. Some CPC-features were also induced in PL-treated ACs compared to fetal bovine serum (FBS)-control ACs. PL treatment of human articular cartilage activates a stem cell niche responsive to injury. These facts can improve the PL therapeutic efficacy in cartilage applications.

scholarly journals The Releasate of Avascular Cartilage Demonstrates Inherent Pro-Angiogenic Properties In Vitro and In Vivo

Cartilage ◽  
2021 ◽  
pp. 194760352110476
Author(s):  
Yannick Nossin ◽  
Eric Farrell ◽  
Wendy J.L.M. Koevoet ◽  
Frank Datema ◽  
Rodrigo A. Somoza ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Nasal Septum ◽  
Healthy Adult ◽  
Cartilage Explants ◽  
Cartilage Tissue ◽  
Human Articular Cartilage ◽  
Cam Assay ◽  
Related Factors

Objective Cartilage is avascular and numerous studies have identified the presence of single anti- and pro-angiogenic factors in cartilage. To better understand the maintenance hyaline cartilage, we assessed the angiogenic potential of complete cartilage releasate with functional assays in vitro and in vivo. Design We evaluated the gene expression profile of angiogenesis-related factors in healthy adult human articular cartilage with a transcriptome-wide analysis generated by next-generation RNAseq. The effect on angiogenesis of the releasate of cartilage tissue was assessed with a chick chorioallantoic membrane (CAM) assay as well as human umbilical vein endothelial cell (HUVEC) migration and proliferation assays using conditioned media generated from tissue-engineered cartilage derived from human articular and nasal septum chondrocytes as well as explants from bovine articular cartilage and human nasal septum. Experiments were done with triplicate samples of cartilage from 3 different donors. Results RNAseq data of 3 healthy human articular cartilage donors revealed that the majority of known angiogenesis-related factors expressed in healthy adult articular cartilage are pro-angiogenic. The releasate from generated cartilage as well as from tissue explants, demonstrated at least a 3.1-fold increase in HUVEC proliferation and migration indicating a pro-angiogenic effect of cartilage. Finally, the CAM assay demonstrated that cartilage explants can indeed attract vessels; however, their ingrowth was not observed. Conclusion Using multiple approaches, we show that cartilage releasate has an inherent pro-angiogenic capacity. It remains vessel free due to anti-invasive properties associated with the tissue itself.

scholarly journals eNAMPT Is Localised to Areas of Cartilage Damage in Patients with Hip Osteoarthritis and Promotes Cartilage Catabolism and Inflammation

2021 ◽  
Vol 22 (13) ◽  
pp. 6719
Author(s):  
Ashleigh M. Philp ◽  
Sam Butterworth ◽  
Edward T. Davis ◽  
Simon W. Jones
Keyword(s):  
Articular Cartilage ◽  
Inflammatory Cytokines ◽  
Cartilage Damage ◽  
Hip Osteoarthritis ◽  
Matrix Metalloproteases ◽  
Cartilage Explants ◽  
Cartilage Tissue ◽  
Human Articular Cartilage ◽  
Nicotinamide Phosphoribosyltransferase ◽  
Pro Inflammatory Cytokines

Obesity increases the risk of hip osteoarthritis (OA). Recent studies have shown that adipokine extracellular nicotinamide phosphoribosyltransferase (eNAMPT or visfatin) induces the production of IL-6 and matrix metalloproteases (MMPs) in chondrocytes, suggesting it may promote articular cartilage degradation. However, neither the functional effects of extracellular visfatin on human articular cartilage tissue, nor its expression in the joint of hip OA patients of varying BMI, have been reported. Hip OA joint tissues were collected from patients undergoing joint replacement surgery. Cartilage explants were stimulated with recombinant human visfatin. Pro-inflammatory cytokines and MMPs were measured by ELISA and Luminex. Localisation of visfatin expression in cartilage tissue was determined by immunohistochemistry. Cartilage matrix degradation was determined by quantifying proteoglycan release. Expression of visfatin was elevated in the synovial tissue of hip OA patients who were obese, and was co-localised with MMP-13 in areas of cartilage damage. Visfatin promoted the degradation of hip OA cartilage proteoglycan and induced the production of pro-inflammatory cytokines (IL-6, MCP-1, CCL20, and CCL4) and MMPs. The elevated expression of visfatin in the obese hip OA joint, and its functional effects on hip cartilage tissue, suggests it plays a central role in the loss of cartilage integrity in obese patients with hip OA.

scholarly journals Repair of full-thickness articular cartilage defects using IEIK13 self-assembling peptide hydrogel in a non-human primate model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexandre Dufour ◽  
Jérôme E. Lafont ◽  
Marie Buffier ◽  
Michaël Verset ◽  
Angéline Cohendet ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Articular Chondrocytes ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Full Thickness ◽  
Primate Model ◽  
Self Assembling ◽  
Articular Cartilage Defects ◽  
Human Primate

AbstractArticular cartilage is built by chondrocytes which become less active with age. This declining function of the chondrocytes, together with the avascular nature of the cartilage, impedes the spontaneous healing of chondral injuries. These lesions can progress to more serious degenerative articular conditions as in the case of osteoarthritis. As no efficient cure for cartilage lesions exist yet, cartilage tissue engineering has emerged as a promising method aiming at repairing joint defects and restoring articular function. In the present work, we investigated if a new self-assembling peptide (referred as IEIK13), combined with articular chondrocytes treated with a chondrogenic cocktail (BMP-2, insulin and T3, designated BIT) could be efficient to restore full-thickness cartilage defects induced in the femoral condyles of a non-human primate model, the cynomolgus monkey. First, in vitro molecular studies indicated that IEIK13 was efficient to support production of cartilage by monkey articular chondrocytes treated with BIT. In vivo, cartilage implant integration was monitored non-invasively by contrast-enhanced micro-computed tomography, and then by post-mortem histological analysis and immunohistochemical staining of the condyles collected 3 months post-implantation. Our results revealed that the full-thickness cartilage injuries treated with either IEIK13 implants loaded with or devoid of chondrocytes showed similar cartilage-characteristic regeneration. This pilot study demonstrates that IEIK13 can be used as a valuable scaffold to support the in vitro activity of articular chondrocytes and the repair of articular cartilage defects, when implanted alone or with chondrocytes.

scholarly journals A scaffold-free approach to cartilage tissue generation using human embryonic stem cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Lauren A. Griffith ◽  
Katherine M. Arnold ◽  
Bram G. Sengers ◽  
Rahul S. Tare ◽  
Franchesca D. Houghton
Keyword(s):  
Stem Cells ◽  
Articular Cartilage ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Articular Chondrocytes ◽  
Embryonic Stem ◽  
Cartilage Tissue ◽  
Human Articular Cartilage ◽  
3D Scaffold ◽  
Tissue Constructs

AbstractArticular cartilage functions as a shock absorber and facilitates the free movement of joints. Currently, there are no therapeutic drugs that promote the healing of damaged articular cartilage. Limitations associated with the two clinically relevant cell populations, human articular chondrocytes and mesenchymal stem cells, necessitate finding an alternative cell source for cartilage repair. Human embryonic stem cells (hESCs) provide a readily accessible population of self-renewing, pluripotent cells with perceived immunoprivileged properties for cartilage generation. We have developed a robust method to generate 3D, scaffold-free, hyaline cartilage tissue constructs from hESCs that are composed of numerous chondrocytes in lacunae, embedded in an extracellular matrix containing Type II collagen, sulphated glycosaminoglycans and Aggrecan. The elastic (Young’s) modulus of the hESC-derived cartilage tissue constructs (0.91 ± 0.08 MPa) was comparable to full-thickness human articular cartilage (0.87 ± 0.09 MPa). Moreover, we have successfully scaled up the size of the scaffold-free, 3D hESC-derived cartilage tissue constructs to between 4.5 mm and 6 mm, thus enhancing their suitability for clinical application.

Genipin Cross-Linked Fibrin Hydrogels for in vitro Human Articular Cartilage Tissue-Engineered Regeneration

2009 ◽  
Vol 190 (6) ◽  
pp. 313-325 ◽  
Author(s):  
Emma V. Dare ◽  
May Griffith ◽  
Philippe Poitras ◽  
James A. Kaupp ◽  
Stephen D. Waldman ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cartilage Tissue ◽  
Human Articular Cartilage

scholarly journals Regenerative Medicine: where their origin makes species meet

2019 ◽  
Vol 37 (3) ◽  
pp. 6-7
Author(s):  
Jos Malda
Keyword(s):  
Articular Cartilage ◽  
Ex Vivo ◽  
Mammalian Species ◽  
Cartilage Tissue ◽  
In Vitro Assays ◽  
Cartilage Defects ◽  
In Vivo Models ◽  
Societal Pressure

The articular cartilage of joints serves diverse functions, including absorbing shock, transmitting force, and enabling low-friction joint motion. Regeneration of articular cartilage defects remains, however, a significant challenge in both human and veterinary orthopaedic practice. Ex vivo and in vivo models play a crucial role in translating novel potential regenerative treatments from bench to bedside. However, in view of the predictive power of these models and the One Medicine concept that proclaim that there should be no dividing lines between human and animal medicine to learn, it is important to understand the similarities as well as differences in the cartilage tissue between species. To this aim, osteochondral cores of the femoral condyles were studied in 58 different mammalian species ranging from mouse to elephant. Interestingly, while biochemical composition remained relatively constant, cartilage thickness and cellularity were similar underscoring the importance of the equine species as a model for human orthopaedic interventions. Nevertheless, political ambition and societal pressure are now asking for a drastically reduction of animal experimentation and have further spiked the development of more predictive in vitro and ex vivo models. In addition to a range of more sophisticated in vitro assays, this has now also provided an ex vivo osteochondral defect model that can be generated based on equine donor tissue. Although such models better represent the situation in the native tissue and can be used to assess (osteo)chondral repair strategies, they still lack some important aspects of the in vivo (patho)physiological (inflammatory) environment, as well as the exposure to mechanical loading. This illustrates the challenges that we still face in translating these novel approaches from bench to bedside.

Human Tibiofemoral Joint Displacements Determined by Displacement-Encoded MRI

2011 ◽  
Author(s):  
D. D. Chan ◽  
C. P. Neu
Keyword(s):  
Articular Cartilage ◽  
Soft Tissues ◽  
The United States ◽  
Tissue Level ◽  
Quantitative Mri ◽  
Cartilage Explants ◽  
Cartilage Tissue ◽  
Human Articular Cartilage ◽  
Tibiofemoral Joint ◽  
Displacement Patterns

Articular cartilage and surrounding soft tissues in the knee are important to normal joint function. Osteoarthritis (OA) is highly prevalent in the United States [1] and features precocious degeneration of articular cartilage. Effective OA treatments require the ability to detect early degeneration, including mechanical and biochemical changes. Magnetic resonance imaging has shown promise for the detection of early degenerative changes, including various quantitative MRI techniques [2]. Displacement-encoded MRI has the ability to detect changes in mechanical behavior, and such techniques have previously been used in cartilage explants [3] and intact juvenile animal joints [4]. However, the authors are aware of no studies with displacement-encoded MRI of human articular cartilage. Tissue-level displacement patterns could be key to revealing early degeneration in articular cartilage. This study demonstrates for the first time displacement encoding with stimulated echoes (DENSE) in an adult human tibiofemoral joint.

scholarly journals In Vitro Effects of Bupivacaine on the Viability and Mechanics of Native and Engineered Cartilage Grafts

2021 ◽  
pp. 036354652199518
Author(s):  
Sarah Oyadomari ◽  
Wendy E. Brown ◽  
Heenam Kwon ◽  
Gaston Otarola ◽  
Jarrett M. Link ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Articular Cartilage ◽  
Cartilage Explants ◽  
Toxic Effects ◽  
Tissue Type ◽  
Cartilage Tissue ◽  
Cartilage Grafts ◽  
Engineer Cartilage ◽  
In Vitro Effects

Background: Although the toxic effects of bupivacaine on chondrocyte monolayer culture have been well described, its cellular and mechanical effects on native and engineered articular cartilage remain unclear. For the repair of articular cartilage defects, fresh autologous and allogenic cartilage grafts are commonly used, and engineered cell-based therapies are emerging. The outcome of grafting therapies aimed at repairing damaged cartilage relies largely on maintaining proper viability and mechanical suitability of the donor tissues. Purpose: To investigate the in vitro effects of single bupivacaine exposure on the viability and mechanics of 2 cartilage graft types: native articular cartilage and engineered neocartilage. Study Design: Controlled laboratory study. Methods: Articular cartilage explants were harvested from the bovine stifle femoral condyles, and neocartilage constructs were engineered from bovine stifle chondrocytes using the self-assembling process, a scaffold-free approach to engineer cartilage tissue. Both explants and neocartilage were exposed to chondrogenic medium containing a clinically applicable bolus of 0.5%, 0.25%, or 0% (control) bupivacaine for 1 hour, followed by fresh medium wash and exchange. Cell viability and matrix content (collagen and glycosaminoglycan) were assessed at t = 24 hours after treatment, and compressive mechanical properties were assessed with creep indentation testing at t = 5 to 6 days after treatment. Results: Single bupivacaine exposure was chondrotoxic in both explants and neocartilage, with 0.5% bupivacaine causing a significant decrease in chondrocyte viability compared with the control condition (55.0% ± 13.4% vs 71.9% ± 13.5%; P < .001). Bupivacaine had no significant effect on matrix content for either tissue type. There was significant weakening of the mechanical properties in the neocartilage when treated with 0.5% bupivacaine compared with control, with decreased aggregate modulus (415.8 ± 155.1 vs 660.3 ± 145.8 kPa; P = .003), decreased shear modulus (143.2 ± 14.0 vs 266.5 ± 89.2 kPa; P = .002), and increased permeability (14.7 ± 8.1 vs 6.6 ± 1.7 × 10−15 m4/Ns; P = .009). Bupivacaine exposure did not have a significant effect on the mechanical properties of native cartilage explants. Conclusion: Single bupivacaine exposure resulted in significant chondrotoxicity in native explants and neocartilage and significant weakening of mechanical properties of neocartilage. The presence of abundant extracellular matrix does not appear to confer any additional resistance to the toxic effects of bupivacaine. Clinical Relevance: Clinicians should be judicious regarding the use of intra-articular bupivacaine in the setting of articular cartilage repair.

Anti-Inflammatory Effects of Novel Standardized Platelet Rich Plasma Releasates on Knee Osteoarthritic Chondrocytes and Cartilage in vitro

2019 ◽  
Vol 20 (11) ◽  
pp. 920-933 ◽  
Author(s):  
Lucía Gato-Calvo ◽  
Tamara Hermida-Gómez ◽  
Cristina R. Romero ◽  
Elena F. Burguera ◽  
Francisco J. Blanco
Keyword(s):  
Gene Expression ◽  
Ex Vivo ◽  
Platelet Rich Plasma ◽  
Cartilage Explants ◽  
Ex Vivo Model ◽  
Chondrocyte Viability ◽  
Platelet Concentration ◽  
The Absolute ◽  
Anti Inflammatory

Background: Platelet Rich Plasma (PRP) has recently emerged as a potential treatment for osteoarthritis (OA), but composition heterogeneity hampers comparison among studies, with the result that definite conclusions on its efficacy have not been reached. Objective: 1) To develop a novel methodology to prepare a series of standardized PRP releasates (PRP-Rs) with known absolute platelet concentrations, and 2) To evaluate the influence of this standardization parameter on the anti-inflammatory properties of these PRP-Rs in an in vitro and an ex vivo model of OA. Methods: A series of PRPs was prepared using the absolute platelet concentration as the standardization parameter. Doses of platelets ranged from 0% (platelet poor plasma, PPP) to 1.5·105 platelets/µl. PRPs were then activated with CaCl2 to obtain releasates (PRP-R). Chondrocytes were stimulated with 10% of each PRP-R in serum-free culture medium for 72 h to assess proliferation and viability. Cells were co-stimulated with interleukin (IL)-1β (5 ng/ml) and 10% of each PRP-R for 48 h to determine the effects on gene expression, secretion and intra-cellular content of common markers associated with inflammation, catabolism and oxidative stress in OA. OA cartilage explants were co-stimulated with IL-1β (5 ng/ml) and 10% of either PRP-R with 0.75·105 platelets/µl or PRP-R with 1.5·105 platelets/µl for 21 days to assess matrix inflammatory degradation. Results: Chondrocyte viability was not affected, and proliferation was dose-dependently increased. The gene expression of all pro-inflammatory mediators was significantly and dose-independently reduced, except for that of IL-1β and IL-8. Immunoblotting corroborated this effect for inducible NO synthase (NOS2). Secreted matrix metalloproteinase-13 (MMP-13) was reduced to almost basal levels by the PRP-R from PPP. Increasing platelet dosage led to progressive loss to this anti-catabolic ability. Safranin O and toluidine blue stains supported the beneficial effect of low platelet dosage on cartilage matrix preservation. Conclusion: We have developed a methodology to prepare PRP releasates using the absolute platelet concentration as the standardization parameter. Using this approach, the composition of the resulting PRP derived product is independent of the donor initial basal platelet count, thereby allowing the evaluation of its effects objectively and reproducibly. In our OA models, PRP-Rs showed antiinflammatory, anti-oxidant and anti-catabolic properties. Platelet enrichment could favor chondrocyte proliferation but is not necessary for the above effects and could even be counter-productive.

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