Demonstration of immunoreactive sites on cartilage after in vivo administration of biotinylated anti-type II collagen antibodies.

Administration of biotinylated monoclonal antibodies provides the basis of a simple technique for identifying immunoreactive sites in vivo. Biotinylated anti-type II collagen antibodies were injected intraperitoneally into normal DBA/1 mice. The mice were sacrificed after 96 hr and the front paws removed and decalcified to allow tissue sectioning before snap-freezing. Binding of antibodies in vivo was visualized with affinity cytochemical staining using avidin-biotin-peroxidase complexes. Specific binding of antibodies to cartilaginous structures was seen after injection of 20-500 micrograms biotinylated monoclonal or polyclonal anti-type II collagen antibodies, but not after injection of a biotinylated control antibody. This technique should further the detection and localization studies of tissue components involved in the dynamics of physiological and pathological processes.

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Prevention of type II collagen-induced arthritis by in vivo treatment with anti-L3T4.

Journal of Experimental Medicine ◽

10.1084/jem.162.3.1105 ◽

1985 ◽

Vol 162 (3) ◽

pp. 1105-1110 ◽

Cited By ~ 254

Author(s):

G E Ranges ◽

S Sriram ◽

S M Cooper

Keyword(s):

T Cells ◽

Lymph Nodes ◽

Humoral Response ◽

Type Ii Collagen ◽

Type Ii ◽

Collagen Induced Arthritis ◽

Delayed Onset ◽

Treatment Administration ◽

In Vivo Administration

The effect of in vivo administration of monoclonal anti-L3T4 antibody on the development of murine collagen-induced arthritis (CIA) was assessed. Treatment with anti-L3T4 resulted in a greater than 90% depletion of L3T4+ T cells in lymph nodes and spleen, an effect that appears entirely reversed 30 d after treatment. Administration of anti-L3T4 before immunization with type II collagen resulted in a significant decrease in arthritis incidence and delayed onset of the disease while treatment begun after a strong anticollagen IgG humoral response was underway was not effective in altering disease expression. These results suggest a prominent role for L3T4+ T cells in the pathogenesis of CIA.

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In vitro and in vivo inhibition of malaria parasite infection by monoclonal antibodies against Plasmodium falciparum circumsporozoite protein (CSP)

Scientific Reports ◽

10.1038/s41598-021-84622-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Merricka C. Livingstone ◽

Alexis A. Bitzer ◽

Alish Giri ◽

Kun Luo ◽

Rajeshwer S. Sankhala ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Monoclonal Antibodies ◽

Disease Burden ◽

Vaccine Candidate ◽

Specific Binding ◽

Circumsporozoite Protein ◽

Target Epitope ◽

Selection Of

AbstractPlasmodium falciparum malaria contributes to a significant global disease burden. Circumsporozoite protein (CSP), the most abundant sporozoite stage antigen, is a prime vaccine candidate. Inhibitory monoclonal antibodies (mAbs) against CSP map to either a short junctional sequence or the central (NPNA)n repeat region. We compared in vitro and in vivo activities of six CSP-specific mAbs derived from human recipients of a recombinant CSP vaccine RTS,S/AS01 (mAbs 317 and 311); an irradiated whole sporozoite vaccine PfSPZ (mAbs CIS43 and MGG4); or individuals exposed to malaria (mAbs 580 and 663). RTS,S mAb 317 that specifically binds the (NPNA)n epitope, had the highest affinity and it elicited the best sterile protection in mice. The most potent inhibitor of sporozoite invasion in vitro was mAb CIS43 which shows dual-specific binding to the junctional sequence and (NPNA)n. In vivo mouse protection was associated with the mAb reactivity to the NANPx6 peptide, the in vitro inhibition of sporozoite invasion activity, and kinetic parameters measured using intact mAbs or their Fab fragments. Buried surface area between mAb and its target epitope was also associated with in vivo protection. Association and disconnects between in vitro and in vivo readouts has important implications for the design and down-selection of the next generation of CSP based interventions.

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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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Monoclonal antibodies to connective tissue macromolecules: Type II collagen

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(80)90352-6 ◽

1980 ◽

Vol 92 (2) ◽

pp. 440-446 ◽

Cited By ~ 113

Author(s):

Thomas F. Linsenmayer ◽

Mary J.C. Hendrix

Keyword(s):

Monoclonal Antibodies ◽

Connective Tissue ◽

Type Ii Collagen ◽

Type Ii

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Induction of erosive arthritis in mice after passive transfer of anti-type II collagen antibodies

Agents and Actions ◽

10.1007/bf01967309 ◽

1989 ◽

Vol 26 (3-4) ◽

pp. 395-395

Author(s):

N. Loutis ◽

P. Bruckner ◽

A. Pataki

Keyword(s):

Type Ii Collagen ◽

Passive Transfer ◽

Type Ii ◽

Erosive Arthritis ◽

Collagen Antibodies

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Discovery and development of a type II collagen neoepitope (TIINE) biomarker for matrix metalloproteinase activity: From in vitro to in vivo

Analytical Biochemistry ◽

10.1016/j.ab.2006.10.034 ◽

2007 ◽

Vol 361 (1) ◽

pp. 93-101 ◽

Cited By ~ 59

Author(s):

O.V. Nemirovskiy ◽

D.R. Dufield ◽

T. Sunyer ◽

P. Aggarwal ◽

D.J. Welsch ◽

...

Keyword(s):

Matrix Metalloproteinase ◽

Type Ii Collagen ◽

Type Ii ◽

Metalloproteinase Activity

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Fucoidan from Ascophyllum nodosum Suppresses Postprandial Hyperglycemia by Inhibiting Na+/Glucose Cotransporter 1 Activity

Marine Drugs ◽

10.3390/md18090485 ◽

2020 ◽

Vol 18 (9) ◽

pp. 485

Author(s):

Xindi Shan ◽

Xueliang Wang ◽

Hao Jiang ◽

Chao Cai ◽

Jiejie Hao ◽

...

Keyword(s):

Glucose Transport ◽

Leptin Receptor ◽

Specific Binding ◽

Fasting Blood Glucose ◽

Postprandial Hyperglycemia ◽

Oral Glucose ◽

Type I ◽

Type Ii ◽

Everted Gut Sac

We previously demonstrated that fucoidan with a type II structure inhibited postprandial hyperglycemia by suppressing glucose uptake, but the mechanism remains elusive. Here, we aimed to assess whether the effect of glucose absorption inhibition was related to the basic structure of fucoidans and preliminarily clarified the underlying mechanism. Fucoidans with type II structure and type I structure were prepared from Ascophyllumnodosum (AnF) or Laminariajaponica (LjF) and Kjellmaniellacrassifolia (KcF), respectively. The effects of various fucoidans on suppressing postprandial hyperglycemia were investigated using in vitro (Caco-2 monolayer model), semi-in vivo (everted gut sac model), and in vivo (oral glucose tolerance test, OGTT) assays. The results showed that only AnF with a type II structure, but not LjF or KcF with type I structure, could inhibit the glucose transport in the Caco-2 monolayer and everted gut sac models. A similar result was seen in the OGTT of Kunming mice and leptin receptor-deficient (db/db) mice, where only AnF could effectively inhibit glucose transport into the bloodstream. Furthermore, AnF (400 mg/kg/d) treatment decreased the fasting blood glucose, HbA1c, and fasting insulin levels, while increasing the serum glucagon-like peptide-1 (GLP-1) level in obese leptin receptor-deficient (db/db) mice. Furthermore, surface plasmon resonance (SPR) analysis revealed the specific binding of AnF to Na+/glucose cotransporter 1 (SGLT1), which indicated the effect of AnF on postprandial hyperglycemia could be due to its suppression on SGLT1 activity. Taken together, this study suggests that AnF with a type II structure can be a promising candidate for hyperglycemia treatment.

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Effects of bio-mimic collagen II-g-hyaluronic acid copolymers on chondrocyte maintenance

Journal of Bioactive and Compatible Polymers ◽

10.1177/0883911519876069 ◽

2019 ◽

Vol 34 (4-5) ◽

pp. 373-385

Author(s):

Kuan Wei Lee ◽

Tang-Ching Kuan ◽

Ming Wei Lee ◽

Chen Show Yang ◽

Lain-Chyr Hwang ◽

...

Keyword(s):

Extracellular Matrix ◽

Cell Proliferation ◽

Hyaluronic Acid ◽

Specific Binding ◽

Type Ii Collagen ◽

Polymerase Chain Reaction Analysis ◽

Collagen Fibrils ◽

Type Ii ◽

Human Cartilage

Extracellular matrix has an important part of the role in tissue engineering and regenerative medicine, so it is necessary to understand the various interactions between cells and extracellular matrix. Type II collagen and hyaluronic acid are the major structural components of the extracellular matrix of articular cartilage, and they are involved in fibril formation, entanglement and binding. The aim of this study was to prepare type II collagen fibrils with surface grafted with hyaluronic acid modified at the reducing end. The topographic pattern of type II collagen fibrils showed a significant change after the surface coupling of hyaluronic acid according to atomic force microscopy scanning. The presence of hyaluronic acid on the type II collagen fibrillar surface was confirmed by the specific binding of nanogold labelled with lectin. No significant increase in cell proliferation was detected by a WST-1 assay. According to histochemical examination, the maintenance of the round shape of chondrocytes and increased glycosaminoglycan secretion revealed that these cell pellets with Col II- g-hyaluronic acid molecules contained un-dedifferentiated chondrocytes in vitro. In the mixture with the 220-kDa Col II- g-hyaluronic acid copolymer, the expression of type II collagen and aggrecan genes in chondrocytes increased as demonstrated by real-time polymerase chain reaction analysis. Experimental results show that the amount of hyaluronic acid added during culturing of chondrocytes can maintain the functionality of chondrocytes and thus allow for increased cell proliferation that is suitable for tissue repair of human cartilage.

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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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