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Highly efficient CRISPR-Cas9-mediated editing identifies novel mechanosensitive microRNA-140 targets in primary human articular chondrocytes.

OBJECTIVE: MicroRNA 140 (miR-140) is a chondrocyte-specific endogenous gene regulator implicated in osteoarthritis (OA). As mechanical injury is a primary aetiological factor in OA, we investigated miR-140-dependent mechanosensitive gene regulation using a novel CRISPR-Cas9 methodology in primary human chondrocytes. METHOD: Primary (passage 1/2) human OA chondrocytes were isolated from arthroplasty samples (six donors) and transfected with ribonuclear protein complexes or plasmids using single guide RNAs (sgRNAs) targeting miR-140, in combination with Cas9 endonuclease. Combinations of sgRNAs and single/double transfections were tested. Gene editing was measured by T7 endonuclease 1 (T7E1) assay. miRNA levels were confirmed by qPCR in chondrocytes and in wild type murine femoral head cartilage after acute injury. Predicted close match off-targets were examined. Mechanosensitive miR-140 target validation was assessed in 42 injury-associated genes using TaqMan Microfluidic cards in targeted and donor-matched control chondrocytes. Identified targets were examined in RNAseq data from costal chondrocytes from miR-140-/- mice. RESULTS: High efficiency gene editing of miR-140 (90-98%) was obtained when two sgRNAs were combined with double RNP-mediated CRISPR-Cas9 transfection. miR-140 levels fell rapidly after femoral cartilage injury. Of the top eight miR-140 gene targets identified (P 

THE CILIARY PROTEIN INTRAFLAGELLAR TRANSPORT 88 IS REQUIRED FOR THE MATURATION, HOMEOSTASIS AND MECHANOADAPTATION OF ARTICULAR CARTILAGE

IS THE CILIARY PROTEIN INTRAFLAGELLAR TRANSPORT 88 A DAMPENER OF MECHANICAL CUES IN ADOLESCENT EPIPHYSEAL PLATE CLOSURE?

RNA-SEQ ANALYSIS OF HAND OSTEOARTHRITIS CARTILAGE REVEALS A RECIPROCAL REGULATION BETWEEN RETINOIC ACID AND MARKERS OF CELL SENESCENCE, IDENTIFYING TALAROZOLE AS A NOVEL TARGETING STRATEGY IN HAND OSTEOARTHRITIS

RESTORING REDOX BALANCE IN CARTILAGE FOLLOWING INJURY RESTORES RETINOIC ACID LEVELS AND PREVENTS MECHANOFLAMMATION

INTRA-ARTICULAR INJECTION OF PHOSPHOLIPID-BASED LUBRICANT REDUCES SHEAR-RESPONSIVE INFLAMMATORY GENES IN THE SUPERFICIAL LAYER OF CARTILAGE POST MURINE JOINT DESTABILISATION

THE MICROBIOME DOES NOT INFLUENCE CARTILAGE DEGRADATION FOLLOWING JOINT DESTABILISATION IN MICE WHEN PERFORMED UNDER STRINGENTLY CONTROLLED CONDITIONS

TNF ALPHA-STIMULATED GENE 6 (TSG6) IS CHONDROPROTECTIVE IN VIVO BUT OVEREXPRESSION CANNOT COMPENSATE FOR LOSS OF FGF2

IS THE PARADOXICAL INCREASE OF CARTILAGE THICKNESS IN CTGF NULL MICE DUE TO COMPENSATION BY ACTIVIN A

Comparison of five-year clinical outcomes of 524 cemented and cementless medial unicompartmental knee replacements.

AIM: To compare the outcomes of cemented and cementless Unicompartmental Knee Replacements (UKR) at 5 years after surgery. METHODS: 262 cemented and 262 cementless medial mobile-bearing UKR, implanted by four high-volume surgeons using identical indications and surgical techniques, were reviewed by independent physiotherapists at 5 years. Survival, Oxford Knee Score (OKS), American Knee Society Score (AKSS), and EQ-5D-5L were assessed. The cementless cohort was mainly implanted after the cemented. Each cohort was divided into early and late sub-groups and compared, to assess if any differences were due to progressive improvement in surgical practice over time. RESULTS: There were no significant differences between the cohorts for demographics, pre-operative scores, and 5-year revision (0.8%), re-operation (1.5%), and complication rates (5%). The cementless cohort had significantly better 5-year OKS (43v41, p = 0.008), AKSS-Objective (94v90, p = 0.049) and EQ-5D-5L (0.81v0.87, p = 0.0001). Pain sub-scores within OKS, AKSS, and EQ-5D-5L were also significantly better in the cementless cohort, and the differences were proportionally much greater and more significant than differences in their respective overall scores. There was no significant improvement in scores between the early and late subgroups of the cohorts, whereas the 'early-cementless' cohort had significantly better scores than the contemporaneously implanted 'late-cemented' cohort. This suggests that differences found were due to implant type, instead of improved surgical practice over time. CONCLUSION: Cementless UKR is associated with better clinical outcomes than cemented UKR, which is primarily due to improved pain relief. Both cemented and cementless UKR are safe with low reoperation and complication rates, and a 5-year survival of 99%.

THE CILIARY PROTEIN INTRAFLAGELLAR TRANSPORT 88 IS REQUIRED FOR THE MATURATION, HOMEOSTASIS AND MECHANOADAPTATION OF ARTICULAR CARTILAGE

IDENTIFICATION OF PUTATIVE SEX-DEPENDENT PAIN PATHWAYS THAT MAY EXPLAIN WHY FEMALE MICE DEVLOP PAIN BEHAVIOUR DESPITE LOWER CHONDROPATHY SCORES

Targeting retinoic acid metabolism as an anti-inflammatory treatment for hand osteoarthritis

THE RELATIONSHIP OF HAND OSTEOARTHRITIS SYMPTOM ONSET WITH MENOPAUSE AND MENOPAUSAL HORMONAL THERAPY-RESULTS OF A RETROSPECTIVE SECONDARY CARE STUDY

Anterior knee pain from the evolutionary perspective.

BACKGROUND: This paper describes the evolutionary changes in morphology and orientation of the PFJ using species present through our ancestry over 340 million years. METHODS: 37 specimens from the Devonian period to modern day were scanned using a 64-slice CT scanner. 3D geometries were created following routine segmentation and anatomical measurements taken from standardised bony landmarks. RESULTS: Findings are described according to gait strategy and age. The adoption of an upright bi-pedal stance caused a dramatic change in the loading of the PFJ which has subsequently led to changes in the arrangement of the PFJ. From Devonian to Miocene periods, our sprawling and climbing ancestors possessed a broad knee with a shallow, centrally located trochlea. A more rounded knee was present from the Paleolithic period onwards in erect and bipedal gait types (aspect ratio 0.93 vs 1.2 in late Devonian), with the PFJ being placed lateral to the midline compared to the medial position in quadrapeds. The depth of the trochlea groove was maximal in the Miocene period of the African ground apes with associated acute sulcus angles in Gorilla (117°) becoming more flattened towards the modern human (138°). CONCLUSIONS: The evolving bipedal gait lead to anteriorisation of the patellofemoral joint, flattening of the trochlea sulcus, in a more lateral, dislocation prone arrangement. Ancestral developments might help explain the variety of presentations of anterior knee pain and patellofemoral instability.

FGF2 promotes regeneration of cartilage in vivo by promoting MSC chemokinesis

Molecular and skeletal characterization of mice with a fibrillin-1 mutation: insight into tissue bioavailability of TGF beta in Marfan syndrome

In a surgical model of OA female mice compared to males display pain at an early stage of disease and differential regulation of pain sensitizers

Tissue sodium controls the release of heparin-binding growth factors, including hepatoma-derived growth factor (HDGF), from cartilage upon injury

Novel target discovery of miR140 targets in human articular chondrocytes