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Copyright
Mesenchymal stem cell encapsulation in alginate
microparticles for intra-articular injection in
osteoarthritis
A. Smith, A. Des Rieux, Mélanie Marquis, Denis Renard, C. Vinatier, J.
Guicheux, C. Le Visage
To cite this version:
A. Smith, A. Des Rieux, Mélanie Marquis, Denis Renard, C. Vinatier, et al.. Mesenchymal stem
cell encapsulation in alginate microparticles for intra-articular injection in osteoarthritis. OASI world
congress on osteoarthritis. Promoting clinical and basic research in osteoarthritis, Apr 2018, Liverpool,
United Kingdom. �10.1016/j.joca.2018.02.588�. �hal-02735209�
547
SAFETY ASSESSMENT OF INTRA-ARTICULAR 2.5% POLYACRYLAMIDE HYDROGEL FOR TREATMENT OF KNEE OSTEOARTHRITIS
A. Overgaard, Sr., M. Henriksen, Sr., H. Bliddal. Frederiksberg-Bispebjerg Hosp., Copenhagen, Denmark
Purpose: There is a drought of available effective treatments of knee osteoarthritis (OA). However, new therapeutics and devices are being introduced, such as polyacrylamide gels. This study was conducted to retrospectively assess the safety and patient-reported treatment effect of intra-articular (IA) treatment of knee OA with a proprietary 2.5% cross-linked polyacrylamide gel (PAAG) in patients with knee OA. Methods: The safety evaluation encompassed a sample of patients who had been treated intra-articularly for knee OA symptoms with 2.5 % pol-yacrylamide hydrogel between March 2010 and February 2017. Safety assessments included the retrospective self-reported adverse events (AEs), medical record reviews, and a clinical examination of the treated knee. If any AEs or discomforts were reported, the patient was asked to describe its duration (“Days”, “Weeks”, “Months”) and severity (“Mild”, “Moderate”, “Severe”). For patients who since treatment had received knee arthroplasty surgery, the surgical records were reviewed for any description of abnormal observations. Patient-reported treatment effect vas measured via a transition questionnaire (Trans-Q), in which the patients scored the current knee status regarding‘pain’, ‘function’, and ‘global’ in relation to pre-treatment status on a 15 points scale from 7 (‘Much worse’) to 7 (‘Much better’), with 0 being ‘No change’. The assessment was done at average 24 months afterfirst treatment (range 4 to 87 months).
Results: Of 126 patients invited to the safety assessment, 91 (72%) accepted the invitation. A majority of the study population (n¼ 66; 73%) reported no adverse events or discomfort after receiving single or mul-tiple treatments with PAAG. Of the 25 patients who reported discomfort or adverse events, 15 reported a sensation of distention immediately after treatment. Most (n¼ 14 93%) described this as passing over a few days to weeks. 10 patients (67%) described this as mild. Two patients sought medical assistance (at the study clinic) after treatment for exacerbations of pain and inflammation, which were treated by oral analgesics and arthrocentesis resolving the symptoms within weeks.
From the clinical examinations, the investigator found no signs of reduced knee ROM or other abnormalities than what is to be expected in a population of knee OA patients. 15 patients had received knee arthroplasty surgery since treatment of which 14 surgical reports were retrieved. In these we found no unexpected descriptions of abnormal-ities that could be associated with prior treatment with PAAG. In fact, none of the reports mention the presence of PAAG, unidentified foreign bodies or otherfindings not expected when inspecting an OA knee. The transition questionnaire describes an overall improvement on pain function and global improvement after treatment with PAAG corre-sponding to a score of 3 (“somewhat better”) for pain, 2 (“A little bet-ter”) for function, and 3 (“somewhat better”) for global. No adverse events causing hospitalizations (serious adverse events), intra-articular infections, or allergic reactions were reported.
Conclusions: This retrospective safety assessment found no significant incidence of adverse events or serious adverse events related to the intra-articular treatment with a proprietary 2.5 % cross-linked poly-acrylamide gel for the relief of knee osteoarthritis pain and disability. The safety of PAAG must be assessed further in a larger prospective study. However, these initial results suggest that the treatment may be regarded as safe.
548
IL4-10 FUSION PROTEIN AS A DISEASE MODIFYING THERAPY FOR OSTEOARTHRITIS EVALUATED IN A RAT GROOVE MODEL IN VIVO J. Popov-Celeketicy, H.M. de Vissery, K. Coeleveldy, C. Steen-Louwsz, C.E. Hackz, H. Weinansx, F.P. Lafebery, S.C. Mastbergeny.
yRheumatology& Clinical Immunology, Univ. Med. Ctr. Utrecht, Utrecht,
Netherlands;zLab. of Translational Immunology, Univ. Med. Ctr. Utrecht, Utrecht, Netherlands; xDept.s of Orthopedics and Rheumatology, Univ. Med. Ctr. Utrecht, Utrecht, Netherlands
Purpose: Osteoarthritis (OA) is a slowly progressive degenerative joint disease. Successful treatment requires the development of a therapy, which would combine the treatments of pain, tissue damage and inflammation, all in one molecule. Fusion protein composed of two anti-inflammatory/regulatory cytokines IL4 and IL10 (IL4-10 FP) was reported
to have chondroprotective, anti-inflammatory and analgesic effects sug-gesting its potential as a disease-modifying OA drug (DMOAD). In order to evaluate efficacy of IL4-10 FP as DMOAD in vivo, we have developed and characterized rat IL4-10 FP and tested it in the rat groove model. Methods: Protein production: rat IL4-10 FP was produced by transient transfection of HEK293F cells and purified by Ni-NTA affinity chroma-tography via hexa-histidine affinity tag, cloned on the N-terminus of the protein. The bioactivity of purified protein was tested in rat whole blood assay. Schematic representation of the in vivo study is shown in Fig. 1. Sixty male Wistar rats were randomly allocated to a standard (n¼ 30) or high-fat diet (n¼ 30). After 12 weeks, OA was induced in 20 rats in each group according to the groove model. Non-grooved rats (n¼ 10 in each dietary group) served as controls. Two weeks after OA induction, rats were treated with weekly intra-articular injections of rat IL4-10 FP (n¼ 20) or PBS (n ¼ 40). IL4-10 FP was administered at a dose of 500 ng per injection per rat. Mechanical hypersensitivity as a measure of pain was evaluated according to von Frey method 24 h before and after intra-articular injections (Fig. 2). Joint degeneration is currently being eval-uated using the OARSI histopathology score for rats.
Results: Intra-articular injections of rat IL4-10 FP significantly reduced pain in rats with induced OA. The transient analgesic effect was observed after intra-articular injections in animals from high-fat diet and stand-ard diet group. In the high-fat diet group, the significant decrease in pain measurement compared to PBS control was observed after the fourth injection and that effect remained till the tenth (end of the experiment) intra-articular injection. In the standard diet group, the significant effect was observed after eight, ninth and tenth intra-articular injection. Conclusions: IL4-10 FP significantly decreased pain in rats with induced OA. The effect was observed after repeated intra-articular injections, however, it was transient. This warrants a more clinically relevant application in order to maintain the beneficial effect for prolonged period. To that end, the rat IL4-10 FP slow release formulations are currently being developed and will be evaluated in the same OA model.
549
MESENCHYMAL STEM CELL ENCAPSULATION IN ALGINATE MICRO-PARTICLES FOR INTRA-ARTICULAR INJECTION IN OSTEOARTHRITIS A. Smithy,z, A. Des Rieuxz,x, M. Marquisk, D. Renardk, C. Vinatiery,
J. Guicheuxy,
¶, C. Le Visagey.yRMeS, Regenerative Med. and Skeleton, Universite de Nantes, ONIRIS, Nantes, France; zLDRI, ADDB, Universite
Catholique de Louvain, Bruxelles, Belgium; xInst. of Condensed Matter
Abstracts / Osteoarthritis and Cartilage 26 (2018) S60eS474 S292
and NanoSci.s, Universite Catholique de Louvain, Louvain-la-Neuve, Belgium; kUR1268 BIA (Biopolymeres Interactions Assemblages), INRA, Nantes, France;¶CHU Nantes, PHU 4 OTONN, Nantes, France
Purpose: Osteoarthritis (OA) is a degenerative and inflammatory joint disease that affects cartilage, subchondral bone and joint tissues. Mes-enchymal Stromal Cells (MSCs) ability to secrete anti-inflammatory and immuno-modulatory factors represents an attractive tool in the treat-ment of osteoarthritis. Considering cell death and the risk of cell leakage upon intra-articular injection, MSCs encapsulation therefore could protect cell from death, avoid cell effusion outside the articular space, and supply a suitable 3D microenvironment supporting the biological activity of MSCs. Previously, we have demonstrated that alginate ticles support MSCs viability and bioactivity. Nevertheless, these par-ticles obtained through a dropwise method were too large to be injected into joints of small animals (1.5± 0.2 mm of diameter). In the present work, we developed a method of cell encapsulation compatible with intra-articular injection through a 26G needle. We also demon-strated cell survival 24 hours after encapsulation and injection. Methods: Human adipose-derived stem cells (hASCs) were encapsu-lated via a micromolding method. We first manufactured poly-dimethylsiloxane (PDMS) chips containing 1600 micromolds with a circular shape, 150mm of diameter and 100mm of depth. The chips were hydrophilized by plasma treatment before their use. For cell encapsu-lation, a solution of 2% alginate (w/v) containing 3 millions of hASCs per mL polymer was deposited onto the chips. The cell loading into the micromolds was done either by sedimentation (10 minutes) or by centrifugation (300 g, 2 min). Alginate particles were then crosslinked with an agarose gel charged with CaCl2 and the encapsulated cells were cultured into 1 mL of complete medium (DMEM, 10% (v/v) FBS, 1% Penicillin/streptomycin). The number of encapsulated cells was eval-uated by a CyQUANT assay immediately and 24 h after encapsulation. To determine the impact of encapsulation on metabolic activity, a presto blue assay was performed 24 hours after encapsulation. We also studied encapsulated cell viability 24 hours after their injection through a 26G needle.
Results: We successfully obtained cylindrical alginate microparticles presenting a diameter of 103± 0.7mm (n¼ 3) and loaded with cells. Using optical microscopy, we noticed that alginate particles contained more cells when loaded by centrifugation than those loaded by sed-imentation. Using cell quantification, we determined that the cen-trifugation method allowed the encapsulation of 30333 (±5552) cells within one chip versus 6056 (±2862) by sedimentation (N ¼ 2; n ¼ 3). This represents 22 (±3) and 4 (±2) cells per particle with centrifugation and sedimentation, respectively. Cell number and metabolic activity remained stable for 24 hours after encapsulation. We also demon-strated that injection through a 26G needle had no impact on the via-bility of encapsulated cells.
Conclusions: Our results show that we are able to encapsulate hASCs into alginate particles by micromolding method. We demonstrated that neither encapsulation by itself nor the method of encapsulation had an impact on cell viability 1 day after encapsulation. Moreover, our par-ticles containing hASCs are injectable through a 26 G needle without affecting cell viability 24 h after injection. Our work will now focus on evaluating in vitro long-term encapsulated cell survival and function-ality in vitro. In case of success, we will then consider intra-articular injection in an animal model of osteoarthritis.
550
RESULTS FROM A 52-WEEK RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED, PHASE 2 STUDY OF A NOVEL, INTRA-ARTICULAR WNT PATHWAY INHIBITOR (SM04690) FOR THE TREATMENT OF KNEE OSTEOARTHRITIS
Y. Yaziciy, T.E. McAlindonz, A. Gibofskyx, N.E. Lanek, D.J. Clauw¶, M.H. Jones#, J. Bergfeld#, C.J. Swearingeny, A. DiFrancescoy, J.R. Tambiahy, M. Hochbergyy.ySamumed, San Diego, CA, USA;zTufts Med. Ctr., Boston, MA, USA;xWeill Cornell Med. and Hosp. for Special Surgery, New York, NY, USA;kUC Davis Med. Ctr., Davis, CA, USA;¶Univ. of Michigan, Ann Arbor, MI, USA;#Cleveland Clinic, Cleveland, OH, USA; yyUniv. of Maryland Sch. of Med., Baltimore, MD, USA
Purpose: Knee osteoarthritis (OA) is characterized by pain, disability and joint deformity due to articular cartilage degradation and bone
remodeling. Wnt signaling is involved in these cellular processes and inflammation. SM04690, a small molecule Wnt pathway inhibitor, is in development as a potential disease modifying OA drug (DMOAD) for knee OA. A phase 2, multicenter, 52-week, randomized, double-blind, placebo-controlled (PBO) trial was conducted to identify a target pop-ulation, determine optimal dose and assess safety of SM04690. The primary endpoint was change from baseline in the target knee in Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) Pain subscore at Week 13.
Methods: Knee OA subjects with Kellgren-Lawrence (KL) grades 2e3, received a single 2 mL injection of 0.03 mg, 0.07 mg, 0.23 mg SM04690 or PBO in target (most painful) knee. WOMAC Pain [0-50] and Function [0e170] were assessed at Weeks 0, 4, 13, 26, 39 and 52, and fixed flexion radiographs (PA, weight-bearing, QuAP™ positioned) were taken at Weeks 0, 26 and 52 for medial joint space width (mJSW). Analysis of covariance adjusted for baseline was conducted with multiple impu-tation in the intention-to-treat (ITT) population. Two subgroups were explored: 1) unilateral symptomatic knee OA subjects as determined by investigator through history and examination (pre-specified) and 2) unilateral symptomatic knee OA subjects without widespread pain (Widespread Pain Index4 and Symptom Severity 2 [WP], post-hoc). Results: 455 subjects (mean age 60.3 [ ± 8.7] years, BMI 29.9 [ ± 4.6] kg/ m2, female 58.9%, KL grade 3 [64.4%], unilateral symptomatic OA
[36.0%]) were enrolled; 91% of patients had bilateral OA on X-ray at Week 0. Serious adverse events, all deemed unrelated to SM04690, were reported in 17 (3.7%) subjects (5 [4.5%, 0.03 mg], 4 [3.5%, 0.07 mg], 4 [3.8%, 0.23 mg], 3 [2.8%, PBO] 1 [6.7%, other]). The primary endpoint was not met. At all timepoints, in the ITT population, clinically mean-ingful improvements in WOMAC Pain and Function (>10% full range) compared to baseline were seen in all groups. At 52 weeks, in the pre-specified unilateral symptomatic subgroup, the 0.07 mg dose group showed significant improvements in WOMAC Pain (P ¼ 0.049) and clinically meaningful and significant improvements in WOMAC Func-tion (P¼ 0.035) compared to PBO.In the post-hoc unilateral sympto-matic without WP subgroup, the 0.07 mg dose group showed both clinically meaningful and significant improvements in WOMAC Pain (P¼ 0.042; P ¼ 0.025) and Function (P ¼ 0.035; P ¼ 0.017) compared to PBO at Weeks 39 and 52, respectively (Fig. 1). Changes in mJSW by treatment group in the total ITT population and the subgroups are shown in Table 1. At 52 weeks, the 0.07 mg unilateral symptomatic (P¼ 0.021) and 0.07 mg unilateral symptomatic without WP (P ¼ 0.032) demonstrated significant increase from baseline in mJSW compared to PBO.
Conclusions: A target population of subjects with unilateral symptoms and potential optimal dose (0.07 mg) of SM04690 were identified. These subjects, especially those without WP, reported significant symptomatic improvements compared to PBO subjects. Significant mJSW improvements compared to PBO were also observed at both 26 and 52 weeks with the 0.07 mg dose group in unilateral symptomatic patients without WP. Therefore, clinical and radiographic outcomes suggest that SM04690 has potential as a DMOAD for knee OA treatment, especially in patients with unilateral symptomatic knee OA without WP. Further clinical studies are ongoing.
