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International audience; Systemic sclerosis (SSc) is a complex autoimmune disease characterized by a functional and structural alteration of the microvascular network associated with cutaneous and visceral fibrosis lesions. Conventional therapies are based on the use of immunomodulatory molecules and symptomatic management but often prove to be insufficient, particularly for patients suffering from severe and rapidly progressive forms of the disease. In this context, cellular therapy approaches could represent a credible solution with the goal to act on the different components of the disease: the immune system, the vascular system and the extracellular matrix. The purpose of this review is to provide an overview of the cellular therapies available for the management of SSc. The first part will focus on systemically injected therapies, whose primary effect is based on immunomodulatory properties and immune system resetting, including autologous hematopoietic stem cell transplantation and intravenous injection of mesenchymal stem cells. The second part will discuss locally administered regenerative cell therapies, mainly derived from adipose tissue, developed for the management of local complications as hand and face disabilities.

The stromal vascular fraction (SVF) is a heterogeneous population of stem/stromal cells isolated from perivascular and extracellular matrix (ECM) of adipose tissue complex (ATC). Administration of SVF holds a strong therapeutic potential for regenerative and wound healing medicine applications aimed at functional restoration of tissues damaged by injuries or chronic diseases. SVF is commonly divided into cellular stromal vascular fraction (cSVF) and tissue stromal vascular fraction (tSVF). Cellular SVF is obtained from ATC by collagenase digestion, incubation/isolation, and pelletized by centrifugation. Enzymatic disaggregation may alter the relevant biological characteristics of adipose tissue, while providing release of complex, multiattachment of cell‐to‐cell and cell‐to‐matrix, effectively eliminating the bioactive ECM and periadventitial attachments. In many countries, the isolation of cellular elements is considered as a “more than minimal” manipulation, and is most often limited to controlled clinical trials and subject to regulatory review. Several alternative, nonenzymatic methods of adipose tissue processing have been developed to obtain via minimal mechanical manipulation an autologous tSVF product intended for delivery, reducing the procedure duration, lowering production costs, decreasing regulatory burden, and shortening the translation into the clinical setting. Ideally, these procedures might allow for the integration of harvesting and processing of adipose tissue for ease of injection, in a single procedure utilizing a nonexpanded cellular product at the point of care, while permitting intraoperative autologous cellular and tissue‐based therapies. Here, we review and discuss the options, advantages, and limitations of the major strategies alternative to enzymatic processing currently developed for minimal manipulation of adipose tissue. stem cells translational medicine 2019;8:1265&1271, Main strategies for adipose tissue processing.

Background Even though the manufacturing processes of the stromal vascular fraction for clinical use are performed in compliance with the good manufacturing practices applying to advanced therapy medicinal products, specifications related to stromal vascular fraction quality remain poorly defined. We analyzed stromal vascular fraction clinical batches from two independent good manufacturing practices-compliant manufacturing facilities, the Swiss Stem Cell Foundation (SSCF) and Marseille University Hospitals (AP-HM), with the goal of defining appropriate and harmonized release acceptance criteria. Methods This retrospective analysis reviewed the biological characteristics of 364 batches of clinical-grade stromal vascular fraction. Collected data included cell viability, recovery yield, cell subset distribution of stromal vascular fraction, and microbiological quality. Results Stromal vascular fraction from SSCF cohort demonstrated a higher viability (89.33% ± 4.30%) and recovery yield (2.54 × 105 ± 1.22 × 105 viable nucleated cells (VNCs) per mL of adipose tissue) than stromal vascular fraction from AP-HM (84.20% ± 5.96% and 2.25 × 105 ± 1.11 × 105 VNCs per mL). AP-HM batches were significantly less contaminated (95.71% of sterile batches versus 74.15% for SSCF batches). The cell subset distribution was significantly different (higher proportion of endothelial cells and lower proportion of leukocytes and pericytes in SSCF cohort). Conclusions Both centers agreed that a good manufacturing practices-compliant stromal vascular fraction batch should exert a viability equal or superior to 80%, a minimum recovery yield of 1.50 × 105 VNCs per mL of adipose tissue, a proportion of adipose-derived stromal cells at least equal to 20%, and a proportion of leukocytes under 50%. In addition, a multiparameter gating strategy for stromal vascular fraction analysis is proposed.

Objective To assess the superiority of adipose tissue-derived stromal vascular fraction (AD-SVF) injection into the fingers vs placebo in reducing hand disability in systemic sclerosis (SSc) patients. Methods We performed a double-blind, multicentre, phase II trial from October 2015 to January 2018 in France. SSc patients with a Cochin Hand Function Scale (CHFS) ≥20/90 were randomized 1:1 to receive injection of AD-SVF or placebo. AD-SVF was obtained using the automated processing Celution 800/CRS system. The placebo was lactated Ringer’s solution. The primary efficacy end point was the change of the CHFS score from baseline to 3 months. Secondary efficacy endpoints included the CHFS score at 6 months, hand function, vasculopathy, hand pain, skin fibrosis, sensitivity of the finger pulps, Scleroderma Health Assessment Questionnaire, patients and physician satisfaction, and safety. Results Forty patients were randomized. The AD-SVF and placebo groups were comparable for age, sex ratio, disease duration, skin fibrosis of the hands and main cause of hand disability. After 3 months’ follow-up, hand function significantly improved in both groups with no between-group difference of CHFS (mean change of −9.2 [12.2] in the AD-SVF group vs −7.6 [13.2] in the placebo group). At 6 months, hand function improved in both groups. Conclusion This study showed an improvement of hand function in both groups over time, with no superiority of the AD-SVF. Considering the limits of this trial, studies on a larger population of patients with homogeneous phenotype and hand handicap should be encouraged to accurately assess the benefit of AD-SVF therapy. Trial registration ClinicalTrials.gov, https://clinicaltrials.gov, NCT02558543. Registered on September 24, 2015.

ObjectiveThe autologous stromal vascular fraction (SVF) from adipose tissue is an alternative to cultured adipose-derived stem cells for use in regenerative medicine and represents a promising therapy for vasculopathy and hand disability in systemic sclerosis (SSc). However, the bioactivity of autologous SVF is not documented in this disease context. This study aimed to compare the molecular and functional profiles of the SVF-based medicinal product obtained from SSc and healthy subjects.MethodsGood manufacturing practice (GMP)-grade SVF from 24 patients with SSc and 12 healthy donors (HD) was analysed by flow cytometry to compare the distribution of the CD45− and CD45+ haematopoietic cell subsets. The ability of SVF to form a vascular network was assessed using Matrigel in vivo assay. The transcriptomic and secretory profiles of the SSc-SVF were assessed by RNA sequencing and multiplex analysis, respectively, and were compared with the HD-SVF.ResultsThe distribution of the leucocyte, endothelial, stromal, pericyte and transitional cell subsets was similar for SSc-SVF and HD-SVF. SSc-SVF retained its vasculogenic capacity, but the density of neovessels formed in SVF-loaded Matrigel implanted in nude mice was slightly decreased compared with HD-SVF. SSc-SVF displayed a differential molecular signature reflecting deregulation of angiogenesis, endothelial activation and fibrosis.ConclusionsOur study provides the first evidence that SSc does not compromise the vascular repair capacity of SVF, supporting its use as an innovative autologous biotherapy. The characterisation of the specific SSc-SVF molecular profile provides new perspectives for delineating markers of the potency of SVF and its targets for the treatment of SSc.

Resume La therapie cellulaire est la discipline par laquelle on peut prevenir, traiter ou attenuer une maladie ou encore reparer un tissu lese par l’injection de cellules humaines. Ces dernieres annees, un nombre croissant de therapies cellulaires innovantes a progresse du stade d’essais cliniques vers la commercialisation et diffusion a grande echelle, et certaines d’entre elles representent des avancees majeures dans des situations en impasse therapeutique. Les outils permettant la mise en œuvre de ces nouvelles therapies sont de plus en plus nombreux et les applications de plus en plus prometteuses.

International audience; Purpose The mechanisms of action and disease-modifying potential of platelet-rich plasma (PRP) injection for osteoarthritis (OA) treatment are still not fully established. The aim of this systematic review of preclinical evidence was to determine if PRP injections can induce disease-modifying effects in OA joints. Methods A systematic review was performed on animal studies evaluating intra-articular PRP injections as treatment for OA joints. A synthesis of the results was performed investigating the disease-modifying effects of PRP by evaluating studies that compared PRP with OA controls or other injectable products, different PRP formulations or injection intervals, and the combination of PRP with other products. The risk of bias was assessed according to the SYRCLE's tool. Results Forty-four articles were included, for a total of 1251 animals. The publication trend remarkably increased over time. PRP injections showed clinical effects in 80% and disease-modifying effects in 68% of the studies, attenuating cartilage damage progression and reducing synovial inflammation, coupled with changes in biomarker levels. Evidence is limited on the best PRP formulation, injection intervals, and synergistic effect with other injectables. The risk of bias was low in 40%, unclear in 56%, and high in 4% of items. Conclusion Intra-articular PRP injections showed disease-modifying effects in most studies, both at the cartilage and synovial level. These findings in animal OA models can play a crucial role in understanding mechanism of action and structural effects of this biological approach. Nevertheless, the overall low quality of the published studies warrants further preclinical studies to confirm the positive findings, as well as high-level human trials to demonstrate if these results translate into disease-modifying effects when PRP is used in the clinical practice to treat OA.

There has been a recent emphasis on the filling capability of fat and also on its regenerative capacity. The stromal vascular fraction of processed fat grafts contains multipotent stem cells that express adipogenic, osteogenic, and chondrogenic genes. Many studies have demonstrated the angiogenic, immunomodulatory, and antiapoptotic capabilities of these cells, making them a valuable and effective tool in restoring devitalized tissue. In this chapter, the authors review the composition and function of adipose tissue in the organism, the isolation process of adipose stromal vascular fraction, its cell composition, and its important angiogenic, immunomodulatory, antifibrotic, and regenerative properties in fat grafting.

International audience; Introduction Platelet-rich plasma (PRP) is able to modulate the joint environment by reducing the inflammatory distress and promoting tissue anabolism. Therefore, it has gained increasing popularity among clinicians in the treatment of osteoarthritis (OA), and it is currently proposed beside consolidated options such as viscosupplementation. Areas covered A systematic review of all available meta-analyses evaluating intra-articular PRP injections in patients affected by knee OA was performed, to understand how this biologic treatment approach compares to the traditional injective therapies available in clinical practice. Moreover, a novel coding system and 'minimum reporting requirements' are proposed to improve future research in this field and promote a better understanding of the mechanisms of action and indications. Expert opinion The main limitation in the current literature is the extreme variability of PRP products used, with often paucity or even lack of data on the biologic features of PRP, which should not be considered as a simple substance, but rather a 'procedure' requiring accurate reporting of the characteristics of the product but also all preparation and application modalities. This approach will aid in matching the optimal PRP product to specific patient factors, leading to improved outcomes and the elucidation of the cost-effectiveness of this treatment.

International audience; Background External radiotherapy has become indispensable in oncological therapies. Unfortunately, radiation is responsible for serious side effects, such as radiodermatitis. The skin is weakened and ulcerated. Our study aimed to evaluate the subcutaneous transfer of microfat (MF) alone and two mixes: MF + Platelet-rich plasma (PRP) and MF + stromal vascular fraction (SVF) to treat radiation-induced skin lesions.Method We defined randomly five experimental groups of nine mice: 1 healthy control group and 4 irradiated (60 Grey) and treated groups. The skin lesions were treated 3 months after irradiation by MF, MF + PRP (50%–50%), MF + SVF (90%–10%) or Ringer-lactate subcutaneous injections. Wound healing was evaluated at 1, 2 and 3 months post-injection and histological wound analysis at 3 months, after euthanasia.Results All the irradiated mice presented with wounds. After sham-injection, the wound area increased by 91.1 ± 71.1% versus a decrease of 15.9 ± 23.1% after MF alone (NS), 27.3 ± 23.8% after MF + SVF (NS) and 76.4 ± 7.7% after MF + PRP (P = 0.032). A significative reduction of skin thickness in wound periphery was measured for the three treated groups compared to sham-injection (P < 0.05) but not in the healed wounds (NS). The most important subcutaneous neo-vessel density was shown after MF + SVF injection.Conclusion The MF + PRP mix was the most efficient product to increase healing. The MF + SVF mix showed the highest rate of neo-angiogenesis but was disappointing in terms of healing.

International audience

Background Vocal fold (VF) scarring, caused by surgery or inflammation, often results in severe voice problems or aphonia. Effective lasting treatment is lacking. Previous in vitro and in vivo animal studies reported positive effects on VF scar resolution with mesenchymal stromal cell (MSC) implantation. The principal aim of this study was to examine safety aspects and secondly treatment efficacy vocal fold function in patients with VF scarring and severe voice problems. Methods In this open-label phase I/II study, 16 patients were treated with surgical scar resection followed by injection of autologous MSCs (0.5–2 × 106 MSCs/patient). Patients were monitored 1 year for serious adverse events (SAE) or minor complications. Therapeutic efficacy on treated VFs was evaluated by measurement of VF vibrations using high-speed laryngoscopy (HSL) and phonation pressure threshold (PTP) for elasticity and VF function. Patients self-reported voice change using the Voice Handicap Index (VHI). Results No SAE or minor side effects were reported. Video ratings of VF vibrations and digitized analysis of HSL and PTP were significantly improved for 62–75% of the patients (depending on parameter). Two patients showed deteriorated VF vibrations, but improved PTP. VHI was significantly improved in 8 patients, with the remaining experiencing no significant change. Conclusions The results indicate that local injection of autologous MSC into scarred VFs with severe voice problems may offer a safe and feasible therapeutic option. VF vibration and elasticity were improved in approximately two thirds of treated patients. This clinical study is registered in clinicaltrials.gov (ID: NCT01981330). Retrospective registration of first patient (20130511). https//: register.clinicaltrials.gov/.

We would like to thank Di Benedetto et al 1 for their constructive comments on our recent publication in Annals of Rheumatic Disease entitled ‘Molecular profile and proangiogenic activity of the adipose-derived stromal vascular fraction used as an autologous innovative medicinal product in patients with systemic sclerosis’.2 Di Benedetto et al mentioned that the profibrotic signature of mesenchymal stem cells (MSCs) derived from patients with the diffuse cutaneous form of systemic sclerosis (dc-SSc) has been established from previous works. In line with this, they discussed the potential risk of increasing the fibrotic burden by using autologous adipose-derived stromal vascular fraction (ADVSF) in which MSCs are highly represented. Although this is a fair question in the context of developing MSCs-based therapy for SSc, we would like to emphasise the fact that the biological findings …

International audience; Purpose: The purpose of this retrospective study was to describe our preliminary results of intrameniscal administration of platelet rich plasma (PRP) in patients with degenerative meniscal tears of the knee. Material and method: Ten patients with degenerative meniscal tears according to the Stoller classification and without knee osteoarthritis were included. There were 7 men and 3 women with a mean age of 40.4 +/- 13.6 [SD] years (range: 18-59 years). Patients were prospectively assessed at baseline and 3- and 6-months after intra meniscal PRP administration. Evaluation included the knee injury and osteoarthritis outcome score (KOOS), pain visual analog scale, and return to competition and training. MRI follow-up was performed 6 months after PRP administration. Adverse events were recorded. Results: Volume of injected PRP was standardized to 4.0 mL. Adverse events during PRP administration was moderate pain in 8 patients (8/10; 80%). Mean KOOS total score significantly improved from 56.6 +/- 15.7 (SD) to 72.7 +/- 18.5 (SD) (P= 0.0007). All six patients practicing sports regularly were able to recover competition or training. In seven patients who underwent MRI follow-up at 6 months, MRI showed stability of the meniscal tears and similar Stoller grades. Conclusion: Intra-meniscal administration of PRP under ultrasound guidance directly into meniscal degenerative lesions is feasible and safe. Further randomized controlled studies are needed to definitely confirm the effectiveness of this procedure. (C) 2019 Societe francaise de radiologie. Published by Elsevier Masson SAS.

Importance Patients with scarred vocal folds, whether congenitally or after phonosurgery, often exhibit dysphonia that negatively affects daily life and is difficult to treat. The autologous adipose tissue–derived stromal vascular fraction (ADSVF) is a readily accessible source of cells with angiogenic, anti-inflammatory, immunomodulatory, and regenerative properties. Objective To evaluate the feasibility and tolerability of local injections of autologous ADSVF in patients with scarred vocal folds. Design, Setting, and Participants CELLCORDES (Innovative Treatment for Scarred Vocal Cords by Local Injection of Autologous Stromal Vascular Fraction) is a prospective, open-label, single-arm, single-center, nonrandomized controlled trial with a 12-month follow-up and patient enrollment from April 1, 2016, to June 30, 2017. Eight patients with severe dysphonia attributable to vocal fold scarring associated with a congenital malformation or resulting from microsurgical sequelae (voice handicap index score >60 of 120) completed the study. Data analysis was performed from September 1, 2018, to January 1, 2019. Interventions Injection of ADSVF into 1 or 2 vocal folds. Main Outcomes and Measures The primary outcomes were feasibility and the number and severity of adverse events associated with ADSVF-based therapy. The secondary outcomes were changes in vocal assessment, videolaryngostroboscopy, self-evaluation of dysphonia, and quality of life at 1, 6, and 12 months after cell therapy. Results Seven women and 1 man (mean [SD] age, 44.6 [10.4] years) were enrolled in this study. Adverse events associated with liposuction and ADSVF injection occurred; most of them resolved spontaneously. One patient received minor treatment to drain local bruising, and another experienced a minor contour defect at the liposuction site. At 12 months, the voice handicap index score was improved in all patients, with a mean (SD) improvement from baseline of 40.1 (21.5) points. Seven patients (88%) were considered to be responders, defined as improvement by 18 points or more in the voice handicap index score (the minimum clinically important difference). Conclusions and Relevance The findings suggest that autologous ADSVF injection in scarred vocal folds is feasible and tolerable. The findings require confirmation in a randomized clinical trial with a larger population. Trial Registration ClinicalTrials.gov Identifier:NCT02622464

International audience; Platelet-rich plasma (PRP) has seen increased interest and utilization over the past decade, particularly in the field of musculoskeletal disease. This growth has been accompanied by the development of medical devices to realize PRP preparation which includes blood collection, centrifugation, and PRP isolation. The final PRP composition is directly influenced by this preparation step and absence of biological quality control led to a lack of comparability between PRP products that could explain the large variability in the clinical benefit of PRP reported in literature. To circumvent this issue, the scientific community developed different PRP classifications but none of them have been adopted. The goal of this review is to furnish both technical and biological characteristics from PRP commercial systems. On review of 1379 studies, 105 studies were selected according to inclusion criteria for technical analysis and led to the identification of 50 commercial systems that have been classified in three technical categories based on the blood harvesting technique (tubes, syringes or bags). Twelve studies were selected and sufficiently describe biological characteristics from only 14 commercial systems from the 50 identified in the technical analysis. Inclusion of duplicates characterization from a same PRP system lead to the final analysis of 36 PRP preparations that met the inclusion criteria of the biological analysis. All these PRP preparations have been classified among the seven existing classifications. Comparison from all biological parameters and classifications revealed a large heterogeneity among the available current PRP commercial systems. Index of biological sensitivity of classifications to distinguish PRP preparations were also variable. Although these findings should help clinicians in selecting a system that meets their specific needs, this also raises the question to standardize the parameters to biologically define PRP preparation among users and to systematically performed PRP qualification when used.

Resume Introduction La secheresse oculaire est une pathologie plurifactorielle de la surface oculaire dont la frequence elevee, le retentissement significatif sur la qualite de vie et sur la vision, ainsi que le cout financier, en font un veritable probleme de sante publique. Si le traitement des formes minimes a moderees est generalement simple et efficace, celui des formes severes est souvent decevant. L’utilisation de collyres a base de serum autologue (CSA) represente une alternative therapeutique envisageable pour les cas les plus severes. L’objectif de notre etude est d’evaluer l’efficacite du traitement par CSA a long-terme chez des patients presentant des syndromes secs isoles refractaires aux traitements conventionnels ou secondaires a d’autres pathologies generales telles que le syndrome de Gougerot-Sjogren et la maladie du greffon contre l’hote (GVH), ou a des pathologies oculaires telles que les keratites neurotrophiques, les brulures chimiques et les pemphigoides cicatricielles oculaires. Patients et methodes Il s’agit d’une etude observationnelle retrospective monocentrique menee sur 47 patients, soit 83 yeux traites par collyre de serum autologue a l’Assistance Publique-Hopitaux de Marseille entre avril 2014 et avril 2017 pour syndrome sec oculaire severe isole ou secondaire. Les symptomes subjectifs des patients (score ocular surface disease index [OSDI]), leur degre de satisfaction ainsi que les effets secondaires ont ete recueillis a l’aide de questionnaires. Le break up time (BUT) et le score de Schirmer ont ete notes. Une evaluation clinique fondee sur un test de coloration corneenne a la fluoresceine (score Oxford) est realisee avant le traitement par CSA a P0 puis selon 5 periodicites : P1 (entre 1 et 3 mois), P2 (3 et 9 mois), P3 (9 a 15 mois) et P4 (15 mois a 24 mois), P5 (> 24 mois). Resultats Sur les 83 yeux traites, l’âge moyen etait de 54,39 ans ± 21,56 dont 20 hommes (42,55 %) et 27 femmes (57,44 %), les indications de traitement comprenaient principalement 25,53 % de GVH, 21,27 % de syndromes secs oculaires severes primitifs et 19,14 % de syndrome de Goujerot-Sjogren primaire. Le suivi moyen a ete de 9,82 mois ± 15,50. La variation du score OSDI entre P0 et P1 montre une diminution de 19,32 points ± 29,37 (p Discussion Des analyses biologiques complementaires sur la composition des CSA sont en cours afin de mettre en evidence des facteurs predictifs d’efficacite. Ainsi, les patients non repondeurs au traitement par CSA pourraient repondre aux collyres de serum allogenique, fabriques a partir de sang peripherique preleve chez un donneur sain a partir de sang de cordon. Conclusion Sur cette premiere serie de 83 yeux traites par CSA, une efficacite clinique a ete notee chez la majorite des patients. Aucune complication infectieuse n’a ete rapportee et le taux de satisfaction a ete tres eleve.

Dear Editor,We read with interest the study published by Paul Harrison and colleagues presenting the characteristics of the leukocytes platelet-rich plasma (L-PRP) preparations for treating Achille...

Resume La microstructure des cordes vocales est complexe et peut etre alteree par une microchirurgie laryngee. On peut alors observer un etat cicatriciel n’autorisant plus le decouplage mecanique entre epithelium et muscle, responsable de difficultes vibratoires a l’origine d’une dysphonie invalidante. Les possibilites therapeutiques actuelles sont reduites et souvent inefficaces sur la vibration : elles n’apportent qu’un effet volumateur limitant la fuite glottique. L’objectif de cette revue de la litterature etait double : i) decrire l’etat actuel de la litterature quant a l’interet de la therapie cellulaire dans la prise en charge des cordes vocales cicatricielles ; ii) analyser l’interet therapeutique de la fraction vasculaire stromale d’origine adipeuse parmi l’arsenal therapeutique preexistant. Notre recherche sur PubMed® en septembre 2016 a reference les articles originaux de langue anglaise ou francaise traitant de l’usage des cellules souches dans la prise en charge des cordes vocales cicatricielles. Vingt-sept articles publies entre 2003 et 2016 repondaient aux criteres de selection. Les cellules souches mesenchymateuses etaient les plus utilisees, majoritairement extraites de la moelle osseuse ou du tissu adipeux. Quatre etudes etaient realisees in vitro sur fibroblastes et dix-huit in vivo chez l’animal. Les criteres d’evaluation etaient : i) l’analyse de la cicatrisation (morphologie macroscopique et microscopique, proprietes viscoelastiques, matrice extracellulaire, fibroblastes) ; ii) l’evaluation de la survie et differenciation des cellules souches. Ces etudes demontrent l’action benefique des cellules souches mesenchymateuses, notamment d’origine adipeuse. Par ailleurs, la fraction vasculaire stromale possede des proprietes qui pourraient permettre d’ameliorer ces resultats, en facilitant la logistique de production.

Vocal fold microstructure is complex and can be affected by laryngeal microsurgery, inducing scarring that prevents mechanical uncoupling of epithelium and muscle, leading to vibration disorder and disabling dysphonia. Treatment options presently are few, and often without efficacy for vibration, having only an impact on volume to reduce glottal closure defect. The present review of the literature had two aims: (i) to report the current state of the literature on cell therapy in vocal fold scarring; and (ii) to analyze the therapeutic interest of the adipose-derived stromal vascular fraction in the existing therapeutic armamentarium. A PubMed® search conducted in September 2016 retrieved English or French-language original articles on the use of stem cells to treat vocal fold scarring. Twenty-seven articles published between 2003 and 2016 met the study selection criteria. Mesenchymal stem cells were most widely used, mainly derived from bone marrow or adipose tissue. Four studies were performed in vitro on fibroblasts, and 18 in vivo on animals. End-points comprised: (i) scar analysis (macro- and micro-scopic morphology, viscoelastic properties, extracellular matrix, fibroblasts); and (ii) assessment of stem cell survival and differentiation. The studies testified to the benefit of mesenchymal stem cells, and especially those of adipose derivation. The stromal vascular fraction exhibits properties that might improve results by facilitating production logistics.

Fat grafting has emerged as a reference procedure in daily plastic surgery practice. Unpredictable fat resorption is the main clinical problem. For this purpose, the addition of PRP to enhance fat revascularization is now an easy and popular procedure. However, no consensus exists regarding the respective volume of fat and PRP used to obtain the ideal mixture. This study investigated the rheological properties of microfat mixed with different proportions of PRP. Results obtained were compared with commercialized hyaluronic acid fillers. Microfat and PRP preparations were performed using standardized techniques. Lipoaspirate residue and blood were obtained from six patients undergoing aesthetic facial microlipofilling. Elastic modulus G’ and tan δ (proportion of elasticity versus fluidity) were obtained for the following conditions: microfat alone and microfat mixed with 10, 30 or 50% of PRP. An expected decrease in elastic modulus was observed by adding increase volumes of PRP. Two groups of products with different rheological properties were considered based on statistical differences highlighted regarding the value of G’. Mean tan δ varied from 0.20 ± 0.04 (microfat alone) to 0.28 ± 0.08 (50% microfat/50% PRP). Microfat mixed with 10% of PRP presents consistency comparable to stiffer fillers, whereas microfat mixed with 30 or 50% corresponds to softer fillers. Rheological differences were highlighted given the proportion of PRP added to the microfat. Further studies assessing the impact of increased doses of platelets in microfat/PRP mixtures on clinical outcomes should also be investigated. Our findings will help clinicians to choose a mixture that meets their specific needs for a given indication. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Introduction Systemic sclerosis (SSc) is a rare autoimmune disease characterized by fibrosis and microvascular damage. Facial signs are frequent and include perioral skin thickening, facial atrophy and microstomia. Facial handicap is overlooked by physicians but when patients are questioned, they frequently declare discomfort and distress due to their facial disability and appearance. As ideal autologous filler, fat graft has previously been considered in the treatment of facial handicap in SSc. To enhance fat graft survival, we combined fat graft and autologous platelet-rich plasma. Cases Description Two patients suffering from SSc with facial disability, facial skin thickening and aesthetic discomfort were treated with combined subcutaneous microinjection of autologous fat and platelet-rich plasma. Conclusions Regular follow-up until 12 months showed that facial disability decreased, skin texture appeared softer, and mouth opening increased. Patients declared to be very satisfied from a functional and aesthetic point of view until their last follow-up at 12 months. Fat grafting is beneficial in the improvement of facial symptoms of patients with SSc, and the addition of platelet-rich plasma is a feasible, simple and safe method. Further studies are required to better assess the advantage on fat graft retention, trophicity and the lasting effect offered by this combined therapy.

Background Platelet-rich plasma improves engraftment after fat transfer. However, the effects of platelet dose have never been investigated. The authors used magnetic resonance imaging to compare surviving graft volumes in mice after administration of four different formulations (microfat alone, and three platelet-rich plasma-enriched microfat mixes). Methods The authors used a random, double-blinded, fat transfer protocol using three different platelet levels: 1 million (low-dose), 500 million (medium-dose), and 1000 million (high-dose) platelets/ml, and fat alone (control). The authors grafted 0.4 ml of the 70/30 platelet-rich plasma-enriched microfat mixtures (0.4 million, 200 million, and 400 million platelets per 0.12 ml for the low-dose, medium-dose, and high-dose mixtures, respectively) or 0.4 ml of microfat alone into 22 nude mice and monitored surviving graft volumes every month for 3 months. Then, the authors histologically analyzed all grafts to assess neoangiogenesis status and fat integrity. Results Three-dimensional magnetic resonance imaging showed that the median surviving graft volumes at 3 months were 9.5 percent (interquartile range, 0 to 25 percent; p = 0.003) (high-dose), 4.1 percent (interquartile range, 0 to 18 percent; p = 0.001) (medium-dose), and 18 percent (interquartile range, 8 to 38 percent; p = 0.41) (low-dose) compared to 36 percent (interquartile range, 28 to 53 percent) for the control value. The histologic integrity of microfat-alone grafts was significantly better than those of the other grafts, although the high-dose and low-dose grafts exhibited higher levels of neoangiogenesis. Conclusion Higher platelet levels in microfat grafts were associated with poor graft survival in nude mice; a clinical review would be appropriate.

Although studies of mixed chimerism following hematopoietic stem cell transplantation in patients with sickle cell disease (SCD) may provide insights into the engraftment needed to correct the disease and into immunological reconstitution, an extensive multilineage analysis is lacking. We analyzed chimerism simultaneously in peripheral erythroid and granulomonocytic precursors/progenitors, highly purified B and T lymphocytes, monocytes, granulocytes and red blood cells (RBC). Thirty-four patients with mixed chimerism and ≥12 months of follow-up were included. A selective advantage of donor RBC and their progenitors/precursors led to full chimerism in mature RBC (despite partial engraftment of other lineages), and resulted in the clinical control of the disease. Six patients with donor chimerism 10 g/dL) and three with AS donors (hemoglobin

International audience

Wrist osteoarthritis (OA) is one of the most common conditions encountered by hand surgeons with limited efficacy of non-surgical treatments. The purpose of this study is to describe the Platelet-Rich Plasma (PRP) mixed-microfat biological characteristics of an experimental Advanced Therapy Medicinal Product (ATMP) needed for clinical trial authorization and describe the clinical results obtained from our first three patients 12 months after treatment (NCT03164122). Biological characterization of microfat, PRP and mixture were analysed in vitro according to validated methods. Patients with stage four OA according to the Kellgren Lawrence classification, with failure to conservative treatment and a persistent daily painful condition &gt, 40 mm according to the visual analog scale (VAS) were treated. Microfat-PRP ATMP is a product with high platelet purity, conserved viability of stromal vascular fraction cells, chondrogenic differentiation capacity in vitro and high secretion of IL-1Ra anti-inflammatory cytokine. For patients, the only side effect was pain at the adipose tissue harvesting sites. Potential efficacy was observed with a pain decrease of over 50% (per VAS score) and the achievement of minimal clinically important differences for DASH and PRWE functional scores at one year in all three patients. Microfat-PRP ATMP presented a good safety profile after an injection in wrist OA. Efficacy trials are necessary to assess whether this innovative strategy could delay the necessity to perform non-conservative surgery.

Developing cartilage constructs with injectability, appropriate matrix composition, and persistent cartilaginous phenotype remains an enduring challenge in cartilage repair. Fourteen patients with minor contour deformity were treated with fluid cartilage filler gently injected as autologous fluid graft in deep planes of defect of the nose that were close to the bone or the cartilage. A computerized tomographic scan control was performed after 12 months. Pearson&rsquo, s Chi-square test was used to investigate differences in cartilage density between native and newly formed cartilages. The endpoints were the possibility of using fluid cartilage as filler with aesthetic and functional improvement and versatility. Patients were followed up for two years. The constructs of fluid cartilage graft that were injected in the deep plane resulted in a persistent cartilage tissue with appropriate morphology, adequate central nutritional perfusion without central necrosis or ossification, and further augmented nasal dorsum without obvious contraction and deformation. This report demonstrated that fluid cartilage grafts are useful for cartilage regeneration in patients with outcomes of rhinoplasty, internal nasal valve collapse, and minor congenital nose aesthetics deformity.

Background/aimsPlatelet-rich plasma (PRP) injections are used in sports medicine and have been the subject of increased clinical interest. However, there have been very few reports of the composition of initial whole blood and the final PRP product. The objective of this study was to provide technical tools to perform a correct characterisation of platelets, leucocytes and red blood cells (RBCs) from whole blood and PRP.MethodsBlood and PRP were obtained from 26 healthy volunteers and prepared according to the varying parameters encountered within PRP process preparation and quantification (harvesting method, anticoagulant used, sampling method, counting method). Concentrations were measured at t=0, t=1, t=6 and t=24 hours.ResultsSampling of blood in Eppendorf tubes significantly decreased platelet concentration over time, whereas sampling in Microvette EDTA-coated tube kept platelet concentration stable until 24 hours. A non-significant difference was observed in platelet counts in PRP with impedance (median (IQR): 521.8 G/L (505.3–524.7)) and fluorescence (591.5 G/L (581.5–595.8)) methods. Other studied parameters did not influence platelet concentrations in blood or PRP samples. Leucocytes and RBC counts were similar whatever the anticoagulant, sampling, harvesting and counting methods used for both blood and PRP samples.ConclusionsSystematic sampling of blood and PRP in EDTA-coated tubes for quality control is recommended. The use of a validated counter for PRP sample should also be taken into account.

International audience; (1) Background: The emergence of injectable “biologic” medication creates a new approach to treat osteoarthritis (OA). Among them, the use of intra-articular injection of PRP became widespread despite the absence of consensus regarding its optimal composition. The aim of this study was to retrospectively correlate an extensive biological characterization of injected PRP to the clinical responses of patients presenting knee OA. (2) Methods: This retrospective study included 75 patients with knee OA. Cartilage lesions were assessed using magnetic resonance imaging and the International Cartilage Regeneration Society (ICRS) classification. PRP extensive biological characterization was performed and patients’ subjective symptoms were recorded before injection and 3 and 6 months after injection using the Knee injury and Osteoarthritis Outcome Score (KOOS). Responders were defined by an improvement of 10 points on KOOS. (3) Results: At 6 months, 63.0% of the patients were responders. Impairment was characterized by a significantly higher proportion of patients with three compartments altered at baseline MRI and receiving a significantly higher dose of platelets compared to responders. (4) Conclusions: Single injection of pure PRP resulted in significant clinical improvement in the management of knee OA. Both baseline MRI and PRP biological features may be predictive factors of the clinical response, highlighting that a better understanding of action mechanism of PRP is still required.

International audience; To evaluate the healing abilities of autologous stem cell therapy (stromal vascular fraction) prepared from adipose tissue we used an automated system without an ex vivo culture phase in a pig model of intrinsic sphincteric deficiency.A total of 15 pigs underwent endoscopic section of the urethral sphincter. Animals were then randomly assigned to 3 groups, including 1) controls without stromal vascular fraction injection, 2) early injection with stromal vascular fraction 2 to 3 days after section and 3) late stromal vascular fraction injection delivery 30 days after injury. Extraction and stromal vascular fraction injection were performed as a single procedure. The stromal vascular fraction was characterized by flow cytometry. Mesenchymal stem cell-like cells were enumerated by clonogenicity (cfu fibroblast) assay. Study end points included histological assessment of the urethral injury surface and urodynamics to determine maximum urethral pressure.Flow cytometry analysis revealed a mesenchymal stem cell-like phenotype in a mean ± SD of 47.3% ± 11.8% of stromal vascular fraction cells. The cfu fibroblast frequency was 1.3 to 6.6/100 stromal vascular fraction cells (1.3% to 6.6%). Stromal vascular fraction injection was associated with a reduction of the urethral injury surface in the early and late injection groups compared with the respective controls (7% vs 17% and 1% vs 13%, p = 0.050 and 0.029, respectively). On day 30 after injection maximum urethral pressure was significantly higher in the injected groups than in the control group, that is 64% vs 50% of maximum urethral pressure on day 0 (p = 0.04).; These data demonstrate the ability of an autologous stromal vascular fraction to improve the urethral healing process in a large animal model of intrinsic sphincteric deficiency.

International audience; Innovative therapies based on autologous adipose-derived stem/stromal cells (ASC) are currently being evaluated for treatment of systemic sclerosis (SSc). Although paracrine angiogenic and antifibrotic effects are considered the predominant mechanisms of ASC therapeutic potential, the impact of SSc on ASC paracrine functions remains controversial. In this study, phenotype, senescence, differentiation potential, and molecular profile were determined in ASC from SSc patients (SSc-ASC) (n = 7) and healthy donors (HD-ASC) (n = 7). ASC were co-cultured in indirect models with dermal fibroblasts (DF) from SSc patients or endothelial cells to assess their pro-angiogenic and antifibrotic paracrine effects. The angiogenic activity of endothelial cells was measured in vitro using tube formation and spheroid assays. DF collagen and alpha smooth muscle actin (αSMA) content were quantified after five days of co-culture with ASC. Differentiation capacity, senescence, and mRNA profiles did not differ significantly between SSc-ASC and HD-ASC. SSc-ASC retained the ability to stimulate angiogenesis through paracrine mechanisms; however, functional assays revealed reduced potential compared to HD-ASC. DF fibrosis markers were significantly decreased after co-culture with SSc-ASC. Together, these results indicate that SSc effects do not significantly compromise the angiogenic and the antifibrotic paracrine properties of ASC, thereby supporting further development of ASC-based autologous therapies for SSc treatment.

International audience; Patients with scarred vocal folds (congenitally or following phonosurgery) are currently difficult to treat and present a dysphonia, often disabling in daily life. Several therapies are available on the market but the results of these are often disappointing. The autologous adipose-derived stromal vascular fraction (ADSVF) is recognized as an easily accessible source of cells with angiogenic, anti-inflammatory, immunomodulatory, and regenerative properties. We present here the case of a 43-year-old woman who had a severe dysphonia associated with scarred vocal folds after a phonosurgery and was resistant to conventional medical and surgical treatments. She received a local administration of autologous ADSVF. The protocol involved, on the same day, adipose tissue extraction, ADSVF preparation, and then local injection (0.45ml of ADSVF in each vocal fold, for a total of 12.2 million ADSVF viable cells). No serious adverse events have been described. One year following the surgery, the laryngoscopic aspect and the majority of voice parameters had improved, in particular the Voice Handicap Index decreasing from 75 to 9. The perceptual analysis found the voice to be less hoarse and more stable, without breathiness. The case of this patient highlights the therapeutic potential of ADSVF for such applications (trial registration, ClinicalTrials.gov NCT02622464; registered 4 December 2015).