Effects of in vivo cyclic compressive loading on the distribution of local type II collagen and superficial lubricin in rat knee articular cartilage

Abstract Background This study aimed to examine the effects of a single episode of in vivo cyclic loading on rat knee articular cartilage (AC) in mid-term observation and investigate relevant factors associated with the progression of post-traumatic osteoarthritis (PTOA). Methods Twelve-week-old Wistar rats underwent one episode of 60 cycles of dynamic compression of 20 N or 50 N on their right knee joint. The spatiotemporal changes in the AC after loading were evaluated using histology and immunohistochemistry at 3 days, 1, 2, 4 and 8 weeks after loading (n=6 for each condition). The chondrocyte vitality was assessed at 1, 3, 6 and 12 hours after loading (n=2 for each condition). ResultsA localized AC lesion on lateral femoral condyle was confirmed in all subjects. The surface and intermediate cartilage in the affected area degenerated after loading, yet the calcified cartilage remained intact. The expression of type II collagen in the lesion cartilage was upregulated after loading, whereas the superficial lubricin layer was eroded in response to cyclic compression. However, the distribution of superficial lubricin gradually recovered to the normal level 4 weeks after loading-induced injury.ConclusionWe confirmed that 60 times cyclic loading exceeding 20 N could result in cartilage damage in rat knee. Endogenous repairs in well-structured joints work well with rebuilding protective layers on the lesion cartilage surface, which could be the latent factor in delaying the progression of PTOA.

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The Complexity of Joint Regeneration: How an Advanced Implant could Fail by Its In Vivo Proven Bone Component

Journal of Trial and Error ◽

10.36850/e3 ◽

2021 ◽

Author(s):

Paweena Diloksumpan ◽

Florencia Abinzano ◽

Mylène de Ruijter ◽

Anneloes Mensinga ◽

Saskia Plomp ◽

...

Keyword(s):

Articular Cartilage ◽

Cartilage Damage ◽

Clinical Signs ◽

Cartilage Regeneration ◽

Type Ii Collagen ◽

Type Ii ◽

Micro Computed Tomography ◽

Bone Anchor

Articular cartilage damage is a major challenge in healthcare due to the lack of long-term repair options. There are several promising regenerative implant-based approaches for the treatment, but the fixation of the implant remains a significant challenge. This study evaluated the potential for repair of an osteochondral implant produced through a novel combined bioprinting-based chondral-bone integration, with and without cells, in an equine model. Implants consisted of a melt electrowritten polycaprolactone (PCL) framework for the chondral compartment, which was firmly integrated with a bone anchor. The bone anchor was produced by extrusion-based printing of a low-temperature setting bioceramic material that had been proven to be effective for osteo-regeneration in an orthotopic, non-load bearing and non-articular site in the same species in an earlier in vivo study. Articular cartilage-derived progenitor cells were seeded into the PCL framework and cultured for 28 days in vitro in the presence of bone morphogenetic protein-9 (BMP-9), resulting in the formation of abundant extracellular matrix rich in glycosaminoglycans (GAGs) and type II collagen. The constructs were implanted in the stifle joints of Shetland ponies with cell-free scaffolds as controls. Clinical signs were monitored, and progression of healing was observed non-invasively through radiographic examinations and quantitative gait analysis. Biochemical and histological analyses 6 months after implantation revealed minimal deposition of GAGs and type II collagen in the chondral compartment of the defect site for both types of implants. Quantitative micro-computed tomography showed collapse of the bone anchor with low volume of mineralized neo-bone formation in both groups. Histology confirmed that the PCL framework within the chondral compartment was still present. It was concluded that the collapse of the osteal anchor, resulting in loss of the mechanical support of the chondral compartment, strongly affected overall outcome, precluding evaluation of the influence of BMP-9 stimulated cells on in vivo cartilage regeneration.

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Intermittent Dynamic Compression Confers Anabolic Effects in Articular Cartilage

Applied Sciences ◽

10.3390/app11167469 ◽

2021 ◽

Vol 11 (16) ◽

pp. 7469

Author(s):

Amalie Engstrøm ◽

Frederik S. Gillesberg ◽

Solveig S. Groen ◽

Peder Frederiksen ◽

Anne-Christine Bay-Jensen ◽

...

Keyword(s):

Articular Cartilage ◽

Mechanical Loading ◽

Kinase Inhibitor ◽

Dynamic Compression ◽

Compressive Loading ◽

Type Ii Collagen ◽

Cartilage Explants ◽

Type Ii ◽

Bovine Articular Cartilage ◽

Collagen Formation

(1) Background: Mechanical loading is an essential part of the function and maintenance of the joint. Despite the importance of intermittent mechanical loading, this factor is rarely considered in preclinical models of cartilage, limiting their translatability. The aim of this study was to investigate the effects of intermittent dynamic compression on the extracellular matrix during long-term culture of bovine cartilage explants. (2) Methods: Bovine articular cartilage explants were cultured for 21 days and subjected to 20 min of 1 Hz cyclic compressive loading five consecutive days each week. Cartilage remodeling was investigated in the presence of IGF-1 or TGF-β1, as well as a TGF-β receptor 1 (ALK5) kinase inhibitor and assessed with biomarkers for type II collagen formation (PRO-C2) and fibronectin degradation (FBN-C). (3) Results: Compression of cartilage explants increased the release of PRO-C2 and FBN-C to the conditioned media and, furthermore, IGF-1 and compression synergistically increased PRO-C2 release. Inhibition of ALK5 blocked PRO-C2 and FBN-C release in dynamically compressed explants. (4) Conclusions: Dynamic compression of cartilage explants increases both type II collagen formation and fibronectin degradation, and IGF-1 interacts synergistically with compression, increasing the overall impact on cartilage formation. These data show that mechanical loading is important to consider in translational cartilage models.

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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.

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Facilitating In Vivo Articular Cartilage Repair by Tissue-Engineered Cartilage Grafts Produced From Auricular Chondrocytes

The American Journal of Sports Medicine ◽

10.1177/0363546517741306 ◽

2017 ◽

Vol 46 (3) ◽

pp. 713-727 ◽

Cited By ~ 7

Author(s):

Chin-Chean Wong ◽

Chih-Hwa Chen ◽

Li-Hsuan Chiu ◽

Yang-Hwei Tsuang ◽

Meng-Yi Bai ◽

...

Keyword(s):

Extracellular Matrix ◽

Articular Cartilage ◽

Cartilage Repair ◽

Articular Chondrocytes ◽

Type Ii Collagen ◽

Type Ii ◽

Articular Cartilage Repair ◽

3 Dimensional ◽

Cell Conversion

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.

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1155. Long-Term Persistence In Vivo of Genetically Engineered Mouse CD4+ T Lymphocytes Targeted to the Articular Cartilage-Specific Protein, Type II Collagen

Molecular Therapy ◽

10.1016/s1525-0016(16)43985-7 ◽

2002 ◽

Vol 5 (5) ◽

pp. S375-S376

Keyword(s):

Articular Cartilage ◽

T Lymphocytes ◽

Type Ii Collagen ◽

Specific Protein ◽

Genetically Engineered ◽

Type Ii

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Method for in vitro production of cartilage microtissues from scaffold-free spheroids composed of human adipose-derived stem cells

Biomedical Research and Therapy ◽

10.15419/bmrat.v7i4.597 ◽

2020 ◽

Vol 7 (4) ◽

pp. 3697-3708

Author(s):

Vy Thi-Kieu Tu ◽

Ha Thi-Ngan Le ◽

Xuan Hoang-Viet To ◽

Phuc Dang-Ngoc Nguyen ◽

Phat Duc Huynh ◽

...

Keyword(s):

Stem Cells ◽

Cartilage Damage ◽

Cartilage Regeneration ◽

Type Ii Collagen ◽

Adipose Derived Stem Cells ◽

Type Ii ◽

In Vitro Production ◽

Engineered Cartilage

Introduction: Cartilage damage is one of the injuries that is difficult for the human body to self-repair due to the avascular and completely mature tissue with only few stem or progenitor cells present. Recently, some studies showed that engineered cartilage tissues could be used to treat or improve this injury. This study aimed to produce the cartilage microtissues from the differentiation of scaffold-free spheroids composed of human adipose-derived stem cells. Methods: Human adipose-derived stem cells (ADSCs) were isolated and expanded following the previously published study. They were then cultured in the non-adherent condition to produce spheroids. The spheroids of the ADSCs were collected and induced into cartilage microtissues in the inducible medium for 21 days. The cartilage microtissue was characterized by some cartilage phenotype markers, including the accumulation of extracellular matrix proteins (aggrecan, glycosaminoglycan, and type II collagen), and the expression of certain genes specific to chondrocytes (Sox9, Col2, Col1, and Acan). Results: The results showed that the expression of chondrocyte-specific genes gradually increased during the 21 days of culture for differentiation. On day 21, the microtissues expressed aggrecan, glycosaminoglycan, and type II collagen proteins. Conclusion: This study demonstrated that cartilage microtissues could easily be produced from scaffold-free spheroids from ADSCs. Thus, cartilage microtissues can be produced in this manner for in vivo transplantation to promote cartilage regeneration, or to produce cartilage tissues for in vitro studies.

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AB0086 Type ii collagen and glycosaminoglycan turnover in bovine articular cartilage is modulated by long-term dynamic compression

10.1136/annrheumdis-2018-eular.6731 ◽

2018 ◽

Author(s):

A. Engstrøm ◽

A.-C. Bay-Jensen ◽

M.A. Karsdal ◽

C.S. Thudium

Keyword(s):

Articular Cartilage ◽

Dynamic Compression ◽

Type Ii Collagen ◽

Type Ii ◽

Bovine Articular Cartilage

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Undenatured Type II Collagen Ameliorates Inflammatory Responses and Articular Cartilage Damage in the Rat Model of Osteoarthritis

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.617789 ◽

2021 ◽

Vol 8 ◽

Author(s):

Cemal Orhan ◽

Vijaya Juturu ◽

Emre Sahin ◽

Mehmet Tuzcu ◽

Ibrahim Hanifi Ozercan ◽

...

Keyword(s):

Articular Cartilage ◽

Inflammatory Responses ◽

Cartilage Damage ◽

Joint Inflammation ◽

Type Ii Collagen ◽

Serum Levels ◽

Joint Disease ◽

Type Ii ◽

Age Related ◽

Undenatured Type Ii Collagen

Osteoarthritis (OA) is an age-related joint disease that includes gradual disruption of the articular cartilage and the resulting pain. The present study was designed to test the effects of undenatured type II collagen (UC-II®) on joint inflammation in the monoiodoacetate (MIA) OA model. We also investigated possible mechanisms underlying these effects. Female Wistar rats were divided into three groups: (i) Control; (ii) MIA-induced rats treated with vehicle; (iii) MIA-induced rats treated with UC-II (4 mg/kg BW). OA was induced in rats by intra-articular injection of MIA (1 mg) after seven days of UC-II treatment. UC-II reduced MIA-induced Kellgren-Lawrence scoring (53.3%, P < 0.05). The serum levels of inflammatory cytokines [IL-1β (7.8%), IL-6 (18.0%), TNF-α (25.9%), COMP (16.4%), CRP (32.4%)] were reduced in UC-II supplemented group (P < 0.0001). In the articular cartilage, UC-II inhibited the production of PGE2 (19.6%) and the expression of IL-1β, IL-6, TNF-a, COX-2, MCP-1, NF-κB, MMP-3, RANKL (P < 0.001). The COL-1 and OPG levels were increased, and MDA decreased in UC-II supplemented rats (P < 0.001). UC-II could be useful to alleviate joint inflammation and pain in OA joints by reducing the expression of inflammatory mediators.

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