Implantation Of Autologous Cells Research Articles

FRACTIONAL LASER TREATMENT COMBINED WITH DYNAMIC SELF-REGENERATING CARTILAGE FORMS A NEW PARADIGM FOR ARTICULAR CARTILAGE REJUVENATION

Lesions in the joint surface are commonly treated with osteoarticular autograft transfer system (OATS), autologous cell implantation (ACI/MACI), or microfracture. Tissue formed buy the latter commonly results in mechanically inferior fibrocartilage that fails to integrate with the surrounding native cartilage, rather than durable hyaline cartilage. Fractional laser treatment to make sub-millimeter (<500 µm) channels has been employed for tissue regeneration in the skin to facilitate rejuvenation without typical scarring. Additionally, we have pioneered a means to generate articular cartilage matrix from chondrocytes—dynamic Self-Regenerating Cartilage (dSRC). Combining these two approaches by performing fractional laser treatment of the joint cartilage and treating with dSRC is a new paradigm for joint surface restoration. This approach was refined in a series of in vitro experiments and tested in swine knee defects during a 6-month study in 12 swine.dSRC are generated by placing 107 swine knee chondrocytes into sealed 15-mL polypropylene tubes and cultured on a rocker at 40 cycles per minute for 14 days at 37°C. The chondrocytes aggregate and generate new extracellular matrix to form a pellet of dSRC. Channels of approximately 300-500 µm diameter were created by infrared laser ablation in swine cartilage (in vitro) and swine knees (in vivo). The diameter and depth of the ablated channel in the cartilage was controlled by the light delivery parameters (power, spot size, pulse duration) from a fractional 2.94 µm Erbium laser. The specimens were evaluated with histology (H&E, safranin O, toluidine blue) and polarized-sensitive optical coherence tomography for collagen orientation.We can consistently create laser-ablated channels in the swine knee and successfully implant new cartilage from dSRC to generate typical hyaline cartilage in terms of morphology and biochemical properties. The neocartilage integrates with host cartilage in vivo.These findings demonstrate our novel combinatorial approach for articular cartilage rejuvenation.

Mesh-tissue integration of synthetic and biologic meshes in wall surgery: brief state of art.

Many studies show that surgical hernia repair with the use of prosthetic meshes can result in pain, hernia recurrence, contraction and mesh rupture. Numerous experimental studies have been conducted to understand the effect of mesh stiffness, pore size and mesh patterns on mesh biocompatibility. The purpose of this mini review is to present an overview of the contracture, adhesion, tissue regrowth and histological response characteristics of permanent and absorbable mesh. Indeed, the mechanics of mesh-human tissue interaction is poorly understood in the literature. It has been shown that early integration of biological meshes is critical for sustained hernia repair. One of the emerging experimental approaches is to combine cell-based regenerative medicine with mesh materials. Studies in preclinical models show that the use of synthetic and biological meshes with autologous cell implantation improves the biocompatibility of biomaterials, promoting key tissue regeneration processes such as adhesion and vascularisation.

Acellular cartilage matrix biomimetic scaffold with immediate enrichment of autologous bone marrow mononuclear cells to repair articular cartilage defects.

Functional repair of articular cartilage defects is always a great challenge in joint surgery clinically. Tissue engineering strategies that combine autologous cell implantation with three-dimensional scaffolds have proven effective for repairing articular cartilage tissue. However, it faces the problem of cell sources and scaffold materials. Autologous chondrocytes and bone marrow are difficult to popularize clinically due to limited donor sources and low mononuclear cell (MNC) concentrations, respectively. The density gradient centrifugation method can increase the concentration of MNCs in fresh bone marrow by nearly a hundredfold and achieve immediate enrichment. In addition, acellular cartilage matrix (ACM), with good biocompatibility and a cartilage-specific microenvironment, is considered to be an ideal candidate scaffold for cartilage regeneration. In this study, hybrid pigs were used to establish articular cartilage defect models of different sizes to determine the feasibility and maximum scope of application of ACM-based biomimetic scaffolds combined with MNCs for inducing articular cartilage regeneration. Importantly, ACM-based biomimetic scaffolds instantly enriched MNCs could improve the repair effect of articular cartilage defects in situ, which established a new model of articular cartilage regeneration that could be applied immediately and suited for large-scale clinical promotion. The current study significantly improves the repair effect of articular cartilage defects, which provides scientific evidence and detailed insights for future clinical applications of ACM-based biomimetic scaffolds combined with MNCs.

Identification of Candidate Synovial Fluid Biomarkers for the Prediction of Patient Outcome After Microfracture or Osteotomy

Background: Biomarkers are needed to predict clinical outcomes for microfracture and osteotomy surgeries to ensure patients can be better stratified to receive the most appropriate treatment. Purpose: To identify novel biomarker candidates and to investigate the potential of a panel of protein biomarkers for the prediction of clinical outcome after treatment with microfracture or osteotomy. Study Design: Descriptive laboratory study. Methods: To identify novel candidate biomarker proteins, we used label-free quantitation after liquid chromatography–tandem mass spectrometry of dynamic range-compressed synovial fluids (SFs) from individuals who responded excellently or poorly (based on change in Lysholm score) to microfracture (n = 6) or osteotomy (n = 7). Biomarkers that were identified in this proteomic analysis or that relate to osteoarthritis (OA) severity or have predictive value in another early OA therapy (autologous cell implantation) were measured in the SF of 19 and 13 patients before microfracture or osteotomy, respectively, using commercial immunoassays, and were normalized to urea. These were aggrecanase-1 (ADAMTS-4), cartilage oligomeric matrix protein (COMP), hyaluronan (HA), lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), matrix metalloproteinase 1 and 3, soluble CD14, S100 calcium binding protein A13, and 14-3-3 protein theta (YWHAQ). Levels of COMP and HA were also measured in the plasma of these patients. To find predictors of postoperative function, multivariable regression analyses were performed. Results: Proteomic analyses highlighted YWHAQ and LYVE-1 as being differentially abundant between the clinical responders/improvers and nonresponders after microfracture. A linear regression model after backward variable selection could relate preoperative concentrations of SF proteins (HA, YWHAQ, LYVE-1), activity of ADAMTS-4, and patient demographic characteristics (smoker status and sex) with Lysholm score 12 months after microfracture. Further, a generalized linear model with elastic net penalization indicated that lower preoperative activity of ADAMTS-4 in SF, being a nonsmoker, and being younger at the time of operation were indicative of a higher postoperative Lysholm score (improved joint function) after osteotomy surgery. Conclusion: We have identified biomarkers and generated regression models with the potential to predict clinical outcome in patients treated with microfracture or osteotomy of the knee. Clinical Relevance: Candidate protein biomarkers identified in this study have the potential to help determine which patients will be best suited to treatment with microfracture or osteotomy.

Autologous cells derived from different sources and administered using different regimens for 'no-option' critical lower limb ischaemia patients.

Autologous cells derived from different sources and administered using different regimens for 'no-option' critical lower limb ischaemia patients.

Promising Biomolecules.

The osteochondral defect (OD) comprises the articular cartilage and its subchondral bone. The treatment of these lesions remains as one of the most problematic clinical issues, since these defects include different tissues, requiring distinct healing approaches. Among the growing applications of regenerative medicine, clinical articular cartilage repair has been used for two decades, and it is an effective example of translational medicine; one of the most used cell-based repair strategies includes implantation of autologous cells in degradable scaffolds such as alginate, agarose, collagen, chitosan, chondroitin sulfate, cellulose, silk fibroin, hyaluronic acid, and gelatin, among others. Concerning the repair of osteochondral defects, tissue engineering and regenerative medicine started to design single- or bi-phased scaffold constructs, often containing hydroxyapatite-collagen composites, usually used as a bone substitute. Biomolecules such as natural and synthetic have been explored to recreate the cartilage-bone interface through multilayered biomimetic scaffolds. In this chapter, a succinct description about the most relevant natural and synthetic biomolecules used on cartilage and bone repair, describing the procedures to obtain these biomolecules, their chemical structure, common modifications to improve its characteristics, and also their application in the biomedical fields, is given.

Cytoskeletal Priming of Mesenchymal Stem Cells to a Medicinal Phenotype

Mesenchymal stem cell (MSC) therapy is a promising approach for the treatment of cardiovascular disease, demonstrating pronounced trophic, immunomodulatory, and pro-angiogenic activity. However, clinical efficacy has suffered from broad variability, presumably due to cell death upon implantation, and the heterogeneous population of autologous cells. Micropatterning single cells in the same geometry can normalize the phenotype in a population, and variations in subcellular curvature will guide focal adhesion, cytoskeletal organization, and the regulation of distinct epigenetic marks to orchestrate a medicinal secretome. Within 2 days, activated cells show elevated expression of pericyte markers and will recapitulate functional pericyte activity through enhanced association with endothelial cell tubules in co-culture. MSCs are believed to undergo a temporary switch in vivo to an activated state in response to injury; thus, we propose engineering actomyosin contractility after isolation can similarly activate MSCs, which may serve as a general approach to prime a medicinal phenotype for cell-based therapies. Patient-derived mesenchymal stem cells will secrete molecules that promote new vasculature and have demonstrated clinical efficacy as a therapy for treating myocardial infarction. While autologous cell implantation is promising, there is considerable variability in current treatments and controversy over the underlying mechanisms. In this work, we reveal that careful control of actomyosin contractility in mesenchymal stem cells can prime a pericyte state with a medicinal secretome that shows sustained pro-angiogenic activity in vitro and in vivo. Activation of mesenchymal stem cells to a medicinal phenotype may increase the efficacy and reproducibility of cell-based therapies for cardiovascular disease

The Effect of Cell Dose on the Early Magnetic Resonance Morphological Outcomes of Autologous Cell Implantation for Articular Cartilage Defects in the Knee

Background: Although autologous chondrocyte implantation (ACI) has been established as a standard treatment for large full-thickness cartilage defects, the effect of different doses of autologous chondrocyte products on structural outcomes has never been examined. Hypothesis: In ACI, the dose level may have an influence on medium-term magnetic resonance morphological findings after treatment. Study Design: Randomized controlled trial; Level of evidence, 1. Methods: A total of 75 patients who underwent ACI using a pure, autologous, third-generation matrix-associated ACI product were divided into 3 groups representing different doses: 3 to 7 spheroids/cm2, 10 to 30 spheroids/cm2, and 40 to 70 spheroids/cm2. Magnetic resonance imaging was performed at 1.5, 3, 6, and 12 months after ACI and was evaluated by the magnetic resonance observation of cartilage repair tissue (MOCART) score and the Knee injury and Osteoarthritis Outcome Score (KOOS). Results: MOCART scores showed improvements after 3 months, with slight dose dependence, and further improvement after 12 months, although without significant dose dependence. The mean MOCART scores after 3 months (0 = worst, 100 = best) were 59.8, 64.5, and 64.7 for the low-, medium-, and high-dose groups, respectively, and 62.9 for all patients; at 12 months, these were 74.1, 74.5, and 68.8 for the respective dose groups and 72.4 for all patients. Several MOCART items (surface of repair tissue, structure of repair tissue, signal intensity of repair tissue, subchondral bone, and synovitis) showed a more rapid response with the medium and high doses than with the low dose, suggesting a potential dose relationship. No significant correlation between the MOCART (overall and subscores) with clinical outcomes as assessed by the overall KOOS was detected at 3- and 12-month assessments. Conclusion: This study reveals a trend toward earlier recovery after treatment with higher spheroid doses in terms of better defect filling for full-thickness cartilage defects of the knee, while outcomes after 12 months were similar in all dose groups. However, a correlation with clinical outcomes or the failure rate at 1 year after ACI was not found. A longer follow-up will be required for more definite conclusions on the clinical relevance of ACI cell density to be drawn. Registration: NCT01225575 (ClinicalTrials.gov identifier); 2009-016816-20 (EudraCT number).

Regeneration of degenerated urinary sphincter muscles: improved stem cell-based therapies and novel imaging technologies.

Stress urinary incontinence (SUI) is a largely ousted but significant medical, social, and economic problem. Surveys suggest that nowadays approximately 10% of the male and 15% of the female population suffer from urinary incontinence at some stage in their lifetime. In women, two major etiologies contribute to SUI: degeneration of the urethral sphincter muscle controlling the closing mechanism of the bladder outflow and changes in lower pelvic organ position associated with degeneration of connective tissue or with mechanical stress, including obesity and load and tissue injury during pregnancy and delivery. In males, the reduction of the sphincter muscle function is sometimes due to surgical interventions as a consequence of prostate cancer treatment, benign prostate hyperplasia, or of neuropathical origin. Accordingly, for women and men different therapies were developed. In some cases, SUI can be treated by physical exercise, electrophysiological stimulation, and pharmacological interventions. If this fails to improve the situation, surgical interventions are required. In standard procedures, endoprostheses for mechanical support of the weakened tissue or mechanical valves for a bladder outflow control are implanted. In 20% of cases treated, repeat procedures are required as implants yield all sorts of side effects in time. Based on preclinical studies, the application of an advanced therapy medicinal product (ATMP) such as implantation of autologous cells may be a curative and long-lasting therapy for SUI. Cellular therapy could also be an option for men suffering from incontinence caused by injury of the nerves controlling the muscular sphincter system. Here we briefly report on human progenitor cells, especially on mesenchymal stromal cells (MSCs), their expansion and differentiation to smooth muscle or striated muscle cells in vitro, labeling of cells for in vivo imaging, concepts of improved, precise, yet gentle application of cells in muscle tissue, and monitoring of injected cells in situ.

Synergistic effects of autologous cell and hepatocyte growth factor gene therapy for neovascularization in a murine model of hindlimb ischemia

Autologous cell implantation and angiogenic gene therapy have been evaluated in critical limb ischemic patients. Here, we compared the features of these strategies individually and in combination. C57BL/6J mice with ischemic hindlimbs were injected with adherent mononuclear cells (aMNCs) from bone marrow or adenovirus encoding the hepatocyte growth factor (HGF) gene (Ad-HGF). Under comparable angiogenic conditions, 10 x 10(5) aMNCs produced significantly higher amounts of VEGF and FGF-2 and stimulated the number of arterioles in ischemic muscle compared with 1 x 10(8) plaque-forming units (pfu) of Ad-HGF. Ad-HGF produced 10 times more HGF in ischemic muscle compared with aMNCs. Injection of 0.3 x 10(5) aMNCs previously transfected with Ad-HGF (aMNC/Ad-HGF) increased blood flow and elevated the numbers of capillaries and arterioles to levels comparable with that seen with 10 x 10(5) aMNCs or 1 x 10(8) pfu of Ad-HGF. Hypoxic conditions induced the apoptotic death of aMNCs. However, coincubation with HGF or aMNC/Ad-HGF protected cells against apoptosis. HGF stimulated the migration of aMNCs, and the migration capacity of the aMNC/Ad-HGF group was significantly higher than that in the aMNC/Ad-LacZ group. In conclusion, cell-based HGF gene therapy decreased the number of cells required for neovascularization. This strategy can be an effective angiogenic therapy.

Principles of cartilage tissue engineering in TMJ reconstruction.

Diseases and defects of the temporomandibular joint (TMJ), compromising the cartilaginous layer of the condyle, impose a significant treatment challenge. Different regeneration approaches, especially surgical interventions at the TMJ's cartilage surface, are established treatment methods in maxillofacial surgery but fail to induce a regeneration ad integrum. Cartilage tissue engineering, in contrast, is a newly introduced treatment option in cartilage reconstruction strategies aimed to heal cartilaginous defects. Because cartilage has a limited capacity for intrinsic repair, and even minor lesions or injuries may lead to progressive damage, biological oriented approaches have gained special interest in cartilage therapy. Cell based cartilage regeneration is suggested to improve cartilage repair or reconstruction therapies. Autologous cell implantation, for example, is the first step as a clinically used cell based regeneration option. More advanced or complex therapeutical options (extracorporeal cartilage engineering, genetic engineering, both under evaluation in pre-clinical investigations) have not reached the level of clinical trials but may be approached in the near future. In order to understand cartilage tissue engineering as a new treatment option, an overview of the biological, engineering, and clinical challenges as well as the inherent constraints of the different treatment modalities are given in this paper.

Marrow stimulation and chondrocyte transplantation using a collagen matrix for cartilage repair

The purpose of the study was to determine whether the implantation of a scaffold would facilitate cartilage repair after microfracture in sheep over a period of 12 months. Furthermore, we investigated the effect of additional autologous cell augmentation of the implanted constructs. Two chondral defects were produced in the medial femoral condyle of sheep without penetrating the subchondral bone. Twenty-seven sheep were divided into the following groups: seven served as untreated controls (Group 1), microfracture was created in 20 animals, seven of them without further treatment (Group 2), in six sheep the defects were additionally covered with a porcine collagen matrix (Group 3), and in seven animals the matrix was augmented with cultured autologous chondrocytes (Group 4). After 4 (11 sheep) and 12 months (16 sheep), the filling of the defects, tissue types, and semiquantitative scores were determined. The untreated defects revealed the least amount of defect fill. Defects treated with microfractures achieved better defect fill, while the additional use of the matrix did not increase the defect fill. The largest quantity of reparative tissue was found in the cell-augmented group. Semiquantitative scores were best in the cell-augmented group. Microfracture treatment was observed to enhance the healing response. The implantation of a cell-seeded matrix further improved the outcome. The implantation of a collagen matrix alone did not enhance repair. Autologous cell implantation appears to be a very important aspect of the tissue engineering approach to cartilage defects.

Current management of cartilage defects: a review

AbstractCartilage defects are a significant cause of morbidity and diminished quality of life, often leading to osteoarthritis (OA). Pain relief, restoration of joint function, and prevention or delay of the onset of OA are the aims of management, and initial treatment is usually conservative but temporary. Recent advances in understanding cartilage biology and the use of diagnostic imaging techniques have, however, allowed more aggressive approaches to cartilage repair which include: arthroscopic lavage with or without debridement; marrow stimulation procedures; implantation of autologous cells or tissue; and knee reconstruction. The role of these techniques in the clinician's effort to match treatment to a patient's needs and expectations is reviewed.

Implantation of autologous cells in minced and devitalized rat skeletal muscles.

Triceps surae of 3 week-old female Wistar rats were minced and orthotopically autografted. Thirty days later, the regenerates developed a force of 314 +/- 58 mN during an isometric maximal tetanus and the maximal area of the muscle tissue was 3.2 +/- 0.5 mm2 (n = 9) in a cross-section. At 60 days, their mean maximal isometric force was increased (1159 +/- 367 mN) as also were their maximal muscle cross-section area (6.8 +/- 0.7 mm2;n = 6). If the minces were devitalized by a freeze and thaw procedure, the regenerates neither contracted under electrical stimulation nor contained any muscle fibre. If devitalized minces were seeded with myogenic cells isolated from the contralateral triceps surae, orthotopically autografted and allowed to regenerate for 30 days, the regeneration was weak: 46 +/- 14 mN of force, 1.2 +/- 0.3 mm2 of muscle area in minces devitalized by cold (n = 7) and 23 +/- 8 mN of force, 0.2 +/- 0.1 mm2 of muscle area in minces devitalized by cold supplemented by heat (n = 8). If viable minces were seeded with myogenic cells, there was no improvement in the extent of regeneration: 275 +/- 83 mN of force, 2.0 +/- 0.6 mm2 of muscle area at 30 days (n = 7) and 738 +/- 191 mN of force, 5.6 +/- 0.9 mm2 of muscle area at 60 days (n = 5). Consequently, although it is possible to induce regeneration by grafting myogenic cells into a devitalized mince, this procedure has no effect when applied to a viable mince.