Leukocytes Cause Inflammatory Response to Traumatized Articular Cartilage in Acute Phase of Joint Injury

Painful and inflamed joints result from joint trauma involving disruption of the cartilage [1]. The pathogenesis of post-traumatic osteoarthritis is not well understood but is most likely multifactorial. Other factors, such as inflammation, may be a critical precursor for post-traumatic arthritis. Transient acute synovitis and inflammation following a traumatic event can persist for months and may be representative of a serious joint injury [2]. Joint effusion aspirates from patients in the acute phase of injury have a higher level of activated leukocytes and an increased rate of reactive oxygen species (ROS) production relative to autologous peripheral blood [3]. In an in vitro study, the presence of inflammatory leukocytes caused more chondrocyte death isolated from traumatized matrix region relative to impacted cartilage alone [4]. Our previous study showed that severe trauma may not be a good predictor for the development of post-traumatic arthritis since chondrocyte death and matrix loss was minimal up to seven days post-trauma [5].

Download Full-text

Cartilage Trauma Induces Necroptotic Chondrocyte Death and Expulsion of Cellular Contents

International Journal of Molecular Sciences ◽

10.3390/ijms21124204 ◽

2020 ◽

Vol 21 (12) ◽

pp. 4204

Author(s):

Josef Stolberg-Stolberg ◽

Meike Sambale ◽

Uwe Hansen ◽

Alexandra Schäfer ◽

Michael Raschke ◽

...

Keyword(s):

Significant Loss ◽

Caspase Inhibitor ◽

Akt Inhibitor ◽

Chondrocyte Death ◽

Transmission Electron ◽

High Prevalence ◽

Post Traumatic ◽

Plasma Membrane Permeabilization ◽

Traumatic Arthritis

Necroptotic cell death is characterized by an activation of RIPK3 and MLKL that leads to plasma membrane permeabilization and the release of immunostimulatory cellular contents. High levels of chondrocyte death occur following intra-articular trauma, which frequently leads to post-traumatic osteoarthritis development. The aim of this study is to assess necroptosis levels in cartilage post-trauma and to examine whether chondrocyte necroptotic mechanisms may be investigated and modified in vitro. Fractured human and murine cartilage, analysed immunohistochemically for necroptosis marker expression, demonstrated significantly higher levels of RIPK3 and phospho-MLKL than uninjured controls. Primary murine chondrocytes stimulated in vitro with the TNFα and AKT-inhibitor alongside the pan-caspase inhibitor Z-VAD-fmk exhibited a significant loss of metabolic activity and viability, accompanied by an increase in MLKL phosphorylation, which was rescued by further treatment of chondrocytes with necrostatin-1. Transmission electron microscopy demonstrated morphological features of necroptosis in chondrocytes following TNFα and Z-VAD-fmk treatment. Release of dsDNA from necroptotic chondrocytes was found to be significantly increased compared to controls. This study demonstrates that cartilage trauma leads to a high prevalence of necroptotic chondrocyte death, which can be induced and inhibited in vitro, indicating that both necroptosis and its consequential release of immunostimulatory cellular contents are potential therapeutic targets in post-traumatic arthritis treatment.

Download Full-text

Acute phase injury of vascular endothelial cells under extra-cellular talaporfin sodium existence: In vitro study

Photodiagnosis and Photodynamic Therapy ◽

10.1016/j.pdpdt.2015.07.104 ◽

2015 ◽

Vol 12 (3) ◽

pp. 351

Author(s):

Risa Hamada ◽

Ryota Matsuzaki ◽

Hiromi Takenoya ◽

Mariko Kurotsu ◽

Emiyu Ogawa ◽

...

Keyword(s):

Endothelial Cells ◽

Acute Phase ◽

Vascular Endothelial Cells ◽

In Vitro Study ◽

Vitro Study ◽

Vascular Endothelial ◽

Talaporfin Sodium

Download Full-text

Targeting pro-inflammatory cytokines following joint injury: acute intra-articular inhibition of interleukin-1 following knee injury prevents post-traumatic arthritis

Arthritis Research & Therapy ◽

10.1186/ar4591 ◽

2014 ◽

Vol 16 (3) ◽

pp. R134 ◽

Cited By ~ 85

Author(s):

Bridgette D Furman ◽

Daniel S Mangiapani ◽

Evan Zeitler ◽

Karsyn N Bailey ◽

Phillip H Horne ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Knee Injury ◽

Interleukin 1 ◽

Joint Injury ◽

Post Traumatic ◽

Traumatic Arthritis ◽

Pro Inflammatory Cytokines

Download Full-text

Overexpression of miR-24 Is Involved in the Formation of Hypocoagulation State after Severe Trauma by Inhibiting the Synthesis of Coagulation Factor X

Disease Markers ◽

10.1155/2017/3649693 ◽

2017 ◽

Vol 2017 ◽

pp. 1-7 ◽

Cited By ~ 8

Author(s):

Lu-Jia Chen ◽

Lian Yang ◽

Xing Cheng ◽

Yin-Kai Xue ◽

Li-Bo Chen

Keyword(s):

Negative Correlation ◽

Major Trauma ◽

Injury Severity ◽

Coagulation Factor ◽

In Vitro Study ◽

Severe Trauma ◽

Trauma Patients ◽

Factor Xii ◽

Factor X

Background. Dysregulation of microRNAs may contribute to the progression of trauma-induced coagulopathy (TIC). We aimed to explore the biological function that miRNA-24-3p (miR-24) might have in coagulation factor deficiency after major trauma and TIC. Methods. 15 healthy volunteers and 36 severe trauma patients (Injury Severity Score ≥ 16 were enrolled. TIC was determined as the initial international normalized ratio >1.5. The miR-24 expression and concentrations of factor X (FX) and factor XII in plasma were measured. In vitro study was conducted on L02 cell line. Results. The plasma miR-24 expression was significantly elevated by 3.17-fold (P=0.043) in major trauma patients and reduced after 3 days (P<0.01). The expression level was significantly higher in TIC than in non-TIC patients (P=0.040). Multivariate analysis showed that the higher miR-24 expression was associated with TIC. The plasma concentration of FX in TIC patients was significantly lower than in the non-TIC ones (P=0.030) and controls (P<0.01). A negative correlation was observed between miR-24 and FX. miR-24 transduction significantly reduced the FX level in the supernatant of L02 cells (P=0.030). Conclusions. miR-24 was overexpressed in major trauma and TIC patients. The negative correlation of miR-24 with FX suggested the possibility that miR-24 might inhibit the synthesis of FX during TIC.

Download Full-text

A VCP modulator, KUS121, as a promising therapeutic agent for post-traumatic osteoarthritis

Scientific Reports ◽

10.1038/s41598-020-77735-2 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Motoo Saito ◽

Kohei Nishitani ◽

Hanako O. Ikeda ◽

Shigeo Yoshida ◽

Sachiko Iwai ◽

...

Keyword(s):

Cell Death ◽

Er Stress ◽

Therapeutic Agent ◽

Articular Chondrocytes ◽

Cartilage Explants ◽

Chondrocyte Death ◽

Post Traumatic ◽

Promising Therapeutic Agent

AbstractPost-traumatic osteoarthritis (PTOA) is a major cause which hinders patients from the recovery after intra-articular injuries or surgeries. Currently, no effective treatment is available. In this study, we showed that inhibition of the acute stage chondrocyte death is a promising strategy to mitigate the development of PTOA. Namely, we examined efficacies of Kyoto University Substance (KUS) 121, a valosin-containing protein modulator, for PTOA as well as its therapeutic mechanisms. In vivo, in a rat PTOA model by cyclic compressive loading, intra-articular treatments of KUS121 significantly improved the modified Mankin scores and reduced damaged-cartilage volumes, as compared to vehicle treatment. Moreover, KUS121 markedly reduced the numbers of TUNEL-, CHOP-, MMP-13-, and ADAMTS-5-positive chondrocytes in the damaged knees. In vitro, KUS121 rescued human articular chondrocytes from tunicamycin-induced cell death, in both monolayer culture and cartilage explants. It also significantly downregulated the protein or gene expression of ER stress markers, proinflammatory cytokines, and extracellular-matrix-degrading enzymes induced by tunicamycin or IL-1β. Collectively, these results demonstrated that KUS121 protected chondrocytes from cell death through the inhibition of excessive ER stress. Therefore, KUS121 would be a new, promising therapeutic agent with a protective effect on the progression of PTOA.

Download Full-text

The effects of radiofrequency energy probe speed and application force on chondrocyte viability

Veterinary and Comparative Orthopaedics and Traumatology ◽

10.1055/s-0037-1616585 ◽

2007 ◽

Vol 20 (01) ◽

pp. 34-37 ◽

Cited By ~ 5

Author(s):

M. L. Meyer ◽

J. J Bogdanske ◽

M. D. Markel ◽

Y. Lu

Keyword(s):

Articular Cartilage ◽

In Vitro Study ◽

Confocal Laser ◽

Radiofrequency Energy ◽

Chondrocyte Viability ◽

Bovine Articular Cartilage ◽

Cell Staining ◽

Chondrocyte Death ◽

Improper Use

Summary Objective: To determine the thermal effects of monopolar radiofrequency energy (mRFE) on bovine articular cartilage when it was moved at different speeds and using varying application forces. Methods: Thirty-six fresh osteochondral sections divided into two groups (18 sections/group) were used in this study. The first group was tested at three speed rates of mRFE probe (1 mm/sec, 5 mm/sec and 10 mm/sec) at a constant force (50 g) applied to the probe tip. In the second group, three application forces of the probe tip were tested (25 g, 50 g and 75 g) at a constant speed (5 mm/sec) (n=6/test). All tests were performed using a custom-built jig to control the mRFE (Vulcan EAS™) probe during a 20-mm pass on each section. After treatment, viability of osteochondral sections was determined by confocal laser microscopy (CLM) combined with vital cell staining. Results: There were not any significant differences in cartilage thickness of tested osteochondral sections among the different speeds or forces. During the mRFE probe treatments at different speeds, CLM demonstrated that probe application at the speed of 1 mm/ sec caused significantly greater chondrocyte death than at the speeds of 5 and 10 mm/sec, whereas there were no significant differences in chondrocyte death among the variable application forces (p>0.05). Discussion: This in vitro study demonstrated that RFE thermal penetration correlated most closely with probe application speed than application force for this mRFE probe. Clinical relevance: Improper use of mRFE may cause thermal injury on articular cartilage.

Download Full-text

Modeling the Early Events of Severe Acute Respiratory Syndrome Coronavirus Infection In Vitro

Journal of Virology ◽

10.1128/jvi.80.6.2684-2693.2006 ◽

2006 ◽

Vol 80 (6) ◽

pp. 2684-2693 ◽

Cited By ~ 81

Author(s):

Yu-Ting Yen ◽

Fang Liao ◽

Cheng-Hsiang Hsiao ◽

Chuan-Liang Kao ◽

Yee-Chun Chen ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Acute Phase ◽

Cellular Response ◽

Immune Cell ◽

A549 Cells ◽

In Vitro Study ◽

Lung Epithelial Cells ◽

Activated T Cells ◽

Monocyte Chemoattractant Protein 1

ABSTRACT The clinical picture of severe acute respiratory syndrome (SARS) is characterized by pulmonary inflammation and respiratory failure, resembling that of acute respiratory distress syndrome. However, the events that lead to the recruitment of leukocytes are poorly understood. To study the cellular response in the acute phase of SARS coronavirus (SARS-CoV)-host cell interaction, we investigated the induction of chemokines, adhesion molecules, and DC-SIGN (dendritic cell-specific ICAM-3-grabbing nonintegrin) by SARS-CoV. Immunohistochemistry revealed neutrophil, macrophage, and CD8 T-cell infiltration in the lung autopsy of a SARS patient who died during the acute phase of illness. Additionally, pneumocytes and macrophages in the patient's lung expressed P-selectin and DC-SIGN. In in vitro study, we showed that the A549 and THP-1 cell lines were susceptible to SARS-CoV. A549 cells produced CCL2/monocyte chemoattractant protein 1 (MCP-1) and CXCL8/interleukin-8 (IL-8) after interaction with SARS-CoV and expressed P-selectin and VCAM-1. Moreover, SARS-CoV induced THP-1 cells to express CCL2/MCP-1, CXCL8/IL-8, CCL3/MIP-1α, CXCL10/IP-10, CCL4/MIP-1β, and CCL5/RANTES, which attracted neutrophils, monocytes, and activated T cells in a chemotaxis assay. We also demonstrated that DC-SIGN was inducible in THP-1 as well as A549 cells after SARS-CoV infection. Our in vitro experiments modeling infection in humans together with the study of a lung biopsy of a patient who died during the early phase of infection demonstrated that SARS-CoV, through a dynamic interaction with lung epithelial cells and monocytic cells, creates an environment conducive for immune cell migration and accumulation that eventually leads to lung injury.

Download Full-text