Autologous Mesenchymal Cells Research Articles

Regulation of Immune Checkpoint Antigen CD276 (B7-H3) on Human Placenta-Derived Mesenchymal Stromal Cells in GMP-Compliant Cell Culture Media.

Therapies utilizing autologous mesenchymal cell delivery are being investigated as anti-inflammatory and regenerative treatments for a broad spectrum of age-related diseases, as well as various chronic and acute pathological conditions. Easily available allogeneic full-term human placenta mesenchymal stromal cells (pMSCs) were used as a potential pro-regenerative, cell-based therapy in degenerative diseases, which could be applied also to elderly individuals. To explore the potential of allogeneic pMSCs transplantation for pro-regenerative applications, such cells were isolated from five different term-placentas, obtained from the dissected maternal, endometrial (mpMSCs), and fetal chorion tissues (fpMSCs), respectively. The proliferation rate of the cells in the culture, as well as their shape, in vitro differentiation potential, and the expression of mesenchymal lineage and stem cell markers, were investigated. Moreover, we studied the expression of immune checkpoint antigen CD276 as a possible modulation of the rejection of transplanted non-HLA-matched homologous or even xeno-transplanted pMSCs. The expression of the cell surface markers was also explored in parallel in the cryosections of the relevant intact placenta tissue samples. The expansion of pMSCs in a clinical-grade medium complemented with 5% human platelet lysate and 5% human serum induced a significant expression of CD276 when compared to mpMSCs expanded in a commercial medium. We suggest that the expansion of mpMSCs, especially in a medium containing platelet lysate, elevated the expression of the immune-regulatory cell surface marker CD276. This may contribute to the immune tolerance towards allogeneic pMSC transplantations in clinical situations and even in xenogenic animal models of human diseases. The endurance of the injected comparably young human-term pMSCs may promote prolonged effects in clinical applications employing non-HLA-matched allogeneic cell therapy for various degenerative disorders, especially in aged adults.

One-stage cartilage repair using the autologous matrix-induced chondrogenesis combined with simultaneous use of autologous adipose tissue graft and adipose tissue mesenchymal cells technique: clinical results and magnetic resonance imaging evaluation at five-year follow-up

PurposeTo evaluate medium-term outcomes of knee cartilage defects repair by autologous matrix-induced chondrogenesis combined with simultaneous use of autologous adipose tissue graft and adipose tissue mesenchymal cells, defined as LIPO-AMIC technique.MethodsThe LIPO-AMIC technique has been used in ICRS degree III–IV knee defects. Eighteen patients have been prospectively evaluated during two and five years both clinically and by MRI.ResultsPatients showed progressive significant improvement of all scores starting early at six months, and further increased values were noted till the last follow-up at 60 months. Mean subjective pre-operative IKDC score of 36.1 significantly increased to 86.4 at 24 months and to 87.2 at 60 months. Mean pre-operative Lysholm score of 44.4 reached 93.5 at two years and 93.5 at five years. MRI examination showed early subchondral lamina regrowth and progressive maturation of repair tissue and filling of defects. The mean total MOCART score showed that a significative improvement from two year follow-up (69.1 points) to last follow-up was 81.9 points (range, 30–100 points, SD 24). Complete filling of the defect at the level of the surrounding cartilage was found in 77.8%.ConclusionsAdipose tissue can represent ideal source of MSCs since easiness of withdrawal and definite chondrogenic capacity. This study clearly demonstrated the LIPO-AMIC technique to be feasible for treatment of knee cartilage defects and to result in statistically significant progressive clinical, functional and pain improvement in all treated patients better than what reported for the AMIC standard technique, starting very early from the 6-month follow-up and maintaining the good clinical results more durably with stable results at mid-term follow-up.

MP42-19 BILAYERED AUTOLOGOUS ADIPOSE-DERIVED MESENCHYMAL CELL-GELATIN SHEETS RECONSTRUCT URETERAL TISSUES IN RABBITS

You have accessJournal of UrologyCME1 Apr 2023MP42-19 BILAYERED AUTOLOGOUS ADIPOSE-DERIVED MESENCHYMAL CELL-GELATIN SHEETS RECONSTRUCT URETERAL TISSUES IN RABBITS Noriyuki Ogawa, Tetsuya Imamura, Tomonori Minagawa, Teruyuki Ogawa, and Osamu Ishizuka Noriyuki OgawaNoriyuki Ogawa More articles by this author , Tetsuya ImamuraTetsuya Imamura More articles by this author , Tomonori MinagawaTomonori Minagawa More articles by this author , Teruyuki OgawaTeruyuki Ogawa More articles by this author , and Osamu IshizukaOsamu Ishizuka More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003280.19AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Treatments of ureteral defects or strictures due to disease or trauma require reconstruction by urological surgery. However, those surgeries have some limitations, such as reoperation, or depending on ureteral length. Tissue engineering techniques would provide novel approaches to avoid the limitations. In this study, we fabricated bilayered autologous adipose-derived mesenchymal cell (AMC)-gelatin sheets and transplanted them into rabbits to replace surgically excised ureteral segments. METHODS: AMCs harvested from abdominal adipose tissues of female New Zealand White rabbits were cultured on collagen-coated dishes and labeled with a red fluorescent dye, PKH26, for later identification. Monolayers of AMCs that were cultured on temperature-responsive dishes were harvested by a gelatin hydrogel sheet. Two AMC-gelatin sheets were then overlaid together with the cell sides juxtaposed, forming a bilayered AMC-gelatin sheet. Following each partial ureterectomy of approximately 1 cm, the bilayered autologous AMC-gelatin sheet was transplanted, joining the proximal and distal ends of the remaining the ureter (experimental group; n=9). Control animals were conducted with a bilayered acellular-gelatin sheet (control group; n=9). RESULTS: At 4 and 8 weeks after transplantation, the proximal regions of ureters of control groups exhibited flexures and dilations compared with experimental groups. Both the control and experimental reconstructed ureteral walls had smooth muscle layers; however, those in the experimental reconstructed ureteral walls were significantly thicker and better organized than those in the control groups. In addition, the lumina of the 8-week reconstructed ureteral tissues in experimental groups did not show histological strictures as seen in the control ureters. Some AMCs differentiated into smooth muscle marker-positive cells. The experimental ureteral walls contained smooth muscle cells derived from the PKH26-labeled AMCs and others that were derived through migration and differentiation of cells from the remaining proximal and distal ends of the original ureter. CONCLUSIONS: These results indicated that the bilayered AMC-gelatin sheets could replace and/or reconstruct ureter tissues. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e575 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Noriyuki Ogawa More articles by this author Tetsuya Imamura More articles by this author Tomonori Minagawa More articles by this author Teruyuki Ogawa More articles by this author Osamu Ishizuka More articles by this author Expand All Advertisement PDF downloadLoading ...

Cell Therapy Based on Gingiva-Derived Mesenchymal Stem Cells Seeded in a Xenogeneic Collagen Matrix for Root Coverage of RT1 Gingival Lesions: An In Vivo Experimental Study

(1) Background: To investigate the effect of a xenogeneic collagen matrix (CMX) seeded with autologous gingiva-derived mesenchymal cells (GMSCs) when combined with a coronally advanced flap (CAF) in the treatment of localized gingival recession type 1 (RT1). (2) Methods: Dehiscence-type defects were created in seven dogs. GMSCs were isolated, transfected with a vector carrying green fluorescent protein (GFP) and expanded. Once chronified, the defects were randomly treated with (1) CAF plus the combination of CMX and GFP+ GMSCs, (2) CAF plus CMX with autologous fibroblasts, (3) CAF plus CMX and (4) CAF alone. Histological and clinical outcomes at 2- and 6-week healing periods were analyzed and compared among groups. (3) Results: Histologically, the addition of autologous cells to the CMX resulted in reduced inflammation and a variable degree of new cementum/bone formation. CMX plus GMSCs resulted in greater mean recession reduction (1.42; SD = 1.88 mm) and percentage of teeth with recession reduction of ≥2 mm (57%) when compared to the other groups, although these differences were not statistically significant. (4) Conclusions: The histometric and clinical results indicated a positive trend favouring the combination of CMX and GMSCs with the CAF when compared to the groups without cells, although these differences were not statistically significant.

Administration of Autologous Mesenchymal Cells for the Treatment of Arthritis.

Injection of autologous mesenchymal stem cells (AMSCs) as stromal vascular fraction, culture expanded adipose derived stem cells, minimally manipulated fat graft, bone marrow aspirate or cultured bone marrow MSCs, for osteo- and inflammatory arthritis have shown good clinical efficacy in many studies. Questions have been raised as to their safety despite no evidence known to us that they are unsafe when used this way. We hypothesized that AMSC injections are completely safe for the treatment of arthritis. A PubMed literature search was performed to identify adverse events specifically related to the injection of autologous mesenchymal or hematopoietic stem cells into arthritic joints or intravenously. 2,011 reported injections were found. No stem cell specific adverse events were identified. Specifically no infections, tumorigenesis, or chondrolysis from collagenase were found. Intra-articular injection of autologous mesenchymal stem cells for the treatment of arthritis is completely safe with no stem cell specific adverse events yet documented, and no increased risk compared with other traditional treatments for arthritis.

No Detectable Alteration of Inorganic Allogeneic Bone Matrix Colonizing Mesenchymal Cells: A Step Towards Personalized Bone Grafts.

BackgroundDuring major bone substance loss, secured allogeneic bone matrix (ABM) is normally utilized for bone repair. Here, we propose a method to colonize ABM using autologous mesenchymal cells (MCs) to improve their integration. Moreover, in this study, the consequences of in vitro colonization on MCs have been evaluated.MethodsAfter in vitro propagation of MCs, their proliferation kinetics on ABM pre-coated with gelatin, fibronectin, collagen IV and human serum (HS) was monitored, and they were compared with cells cultured without ABM for 8 weeks. The effect of ABM on cell phenotype was also assessed. Lastly, the ability of ABM-colonizing MCs to perform hematopoiesis, a function normally preserved in selected culture conditions, and their differentiation towards osteoblastic lineage were evaluated.ResultsMC and colony-forming unit-fibroblast proliferated 930- and 590-fold, respectively. The proliferation rate of the expanded MCs was higher, forming a 3-dimensional structure in all ABMs. Pre-coating with HS was the most efficient treatment of ABMs to increase the initial adherence of MCs, and it partly explains the reason for the higher propagation of MCs. Flow cytometry analyses revealed subtle alterations in ABM-colonizing cells; however, the ability of MCs to maintain long-term culture initiating cells proliferation and differentiate into osteoblastic lineage was preserved.ConclusionsIn this study, the in vitro biocompatibility of bone marrow (BM) MCs with ABMs, the role of HS in scaffold coating, and the possibility of initially using a small BM sample for this approach were demonstrated.

Assessment of immune-alternations and their correlations with therapeutic outcomes of transplantation of autologous Mesenchymal and Allogenic fetal stem cells in patients with type 1 diabetes: a study protocol.

Stem-cell therapy, which has recently emerged as a potentially therapeutic option for diabetes, is demonstrated to significantly alter both cellular and non-cellular elements of the immune system. In addition, it is demonstrated that allogenic stem-cells, once considered immune-privileged, can be rejected by the host immune system almost similar to any other somatic cell. To date, nonetheless, details of these intricate interactions remain obscure. The current study is designed to illuminate both aforementioned favorable and unfavorable stem cell-mediated immune reactions. Findings of this study may shed some light on how stem cells may exert their therapeutic effect in type 1 diabetes through immune system-mediated mechanisms and illuminate the partially-obscure immune-caused rejection of these cells. For the purpose of this study, frozen whole blood samples obtained from patients with type 1 diabetes who received stem cells at the Endocrinology and Metabolism Research Institute of Tehran University of Medical Sciences in two different clinical trials will be thawed and analyzed. These clinical trials were carried out using two different sources of stem cells, namely allogenic fetal and autologous mesenchymal cells. The samples we aim to analyze were obtained from the patients before the procedure and regularly after it, one, three, six, 12, and 24months later. For the purpose of this study, the following parameters will be measured: C-peptide levels, IDAA1c (a surrogate marker of beta cell function which is calculated as HbA1c (%) + [4 × insulin dose (units per kilogram per day)]), frequencies of islet-specific autoreactive CD8+ T cells (CTL), different lymphocyte subsets, thymic function indicators, T cell repertoire diversity (including Treg/Tconv ratios), plasma levels of several pro- and anti-inflammatory cytokines, diabetes autoantibodies, and HLA typing. The stem cell transplantation clinical trials which provided the primary source of our samples were carried out at the Endocrinology and Metabolism Research Institute of Tehran University of Medical Sciences between 2008 and 2012. These series of clinical trials have secured approval of the ethics committee of Tehran University of Medical Sciences (ethical code number: E-0089) and registered on the national clinical trial registry of Islamic Republic of Iran (IRCT) with the identifier codes: IRCT138810271414N8 (for autologous mesenchymal cells) and IRCT201103171414N23 (for allogenic fetal cells). Our findings are to be presented at international scientific events, published in peer-reviewed journals, and disseminated both electronically and in print. Besides, results of the current study will be used for design and implementation of future laboratory investigations and clinical trials at the Endocrinology and Metabolism Research Institute of Tehran University of Medical Sciences.

Characteristics of carrier cells associated with clinical response in children treated with adenoviral virotherapy administered in autologous marrow-derived mesenchymal cells.

e15217 Background: We used oncolytic virotherapy carried by autologous marrow-derived mesenchymal cells to treat children with metastatic/refractory tumors (NCT01844661; 10.1016/j.ymthe.2020.01.019 and 10.1016/j.canlet.2015.11.036). Most patients did not show clinical response while a small group did. Methods: We have carried out a comprehensive study of the transcriptome of this new advanced therapy medicine (ATM) to find determinants that might be associated with clinical outcome. Results: The study of the ATM's transcriptome from a first cohort of patients identified signatures associated to biological processes in the infected carrier cells that distinguished responders from non-responders (viral infection and nucleic acid metabolism; immune regulatory routes; cell-cell adhesion; mitosis; cell metabolism; response to stress; and bone marrow fibrosis). Next, we used an approach based on systems biology and neural networks to identify candidates responsible for the differences found in gene expression signatures. We then validated the identified candidates using samples obtained from a second independent cohort of patients, by quantitative PCR and ELISA. Mesenchymal cells of the responding patients expressed significantly lower levels of the MAVS and NDRG1 genes compared with those of the non-responders after oncolytic adenoviral infection. Both genes are related to cellular antiviral responses and cellular metabolism. Accordingly, mesenchymal cells of responders produced significantly lower amounts of IL-6 and CCL-2 than non-responders’ cells. Conclusions: These results point to possible pretreatment biomarkers and the possibility of optimizing a universal version of the carrier cells for the oncolytic virus in this new ATM.

Systemic oncolytic adenovirus delivered in mesenchymal carrier cells modulate tumor infiltrating immune cells and tumor microenvironment in mice with neuroblastoma.

Celyvir (autologous mesenchymal cells -MSCs- that carry an oncolytic adenovirus) is a new therapeutic strategy for metastatic tumors developed by our research group over the last decade. There are limitations for studying the immune effects of human oncolytic adenoviruses in murine models since these viruses do not replicate naturally in these animals. The use of xenografts in immunodeficient mice prevent assessing important clinical aspects of this therapy such as the antiadenoviral immune response or the possible intratumoral immune changes, both of tumor infiltrating leukocytes and of the microenvironment. In our strategy, the presence of MSCs in the medicinal product adds an extra level of complexity. We present here a murine model that overcomes many of these limitations. We found that carrier cells outcompeted intravenous administration of naked particles in delivering the oncolytic virus into the tumor masses. The protection that MSCs could provide to the oncolytic adenovirus did not preclude the development of an antiadenoviral immune response. However, the presence of circulating antiadenoviral antibodies did not prevent changes detected at the tumor masses: increased infiltration and changes in the quality of immune cells per unit of tumor volume, and a less protumoral and more inflammatory profile of the tumor microenvironment. We believe that the model described here will enable the study of crucial events related to the immune responses affecting both the medicinal product and the tumor.

Design of clinical prospective study: application of gene, cell therapy and indirect revascularization for treatment of patients with non-reconstructive chronic lower limb ischemia

The aim — to improve the results of surgical treatment of patients with chronic critical lower limb ischemia.Materials and methods. To organize the study, we chose the open controlled method of parallel groups. The study was planned and conducted as an open prospective randomized and controlled trial. The study was conducted in four stages: 1st (screening, up to 2 weeks) — evaluation of the possibility of involvement in the study. At this stage all examinations of patients were carried out; 2nd (2 days) — signing informed consent, distribution of the patients to the treatment groups; 3rd (1.5 months) — treatment; 4th (3 years) — observation. The study involved patients whose vascular bed surgical reconstruction was impossible due to: the presence of prolonged occlusion of the femoral artery with the transition to the distal bed; destruction of vessels of distal segments with unfavorable prognosis; previously performed reconstructive interventions on the arteries of the lower extremities, which did not eliminate chronic ischemia due to the impossibility of repeated surgery; decompensation of somatic pathology that could interfere with surgery.Results and discussion. 110 patients were selected for treatment. The patients were divided into three groups according to the treatment method: 1st — 48 patients who underwent revascularization osteotreopenia as an operation of indirect revascularization, 2nd — 21 patients who were administered endothelial cell growth factor drug for stimulation of their own angiogenesis, 3rd — 41 patients in whom subcutaneous fat sampling, cultivation and transplantation of autologous mesenchymal cells were performed according to proposed by us method. To compare the main endpoints of the study, we created a control group of patients, who had previously undergone the treatment by standard methods in the clinic. The criteria for inclusion in the treatment groups and control group did not differ. All patients in the treatment groups had a high level of pain (on average > 70 mm on a visual analog scale) in the affected limb. The most common associated pathologies were hypertension and coronary heart disease. There was no statistically significant difference in the anamnestic data. Many patients had a history of revascularization attempts (> 50 % had one attempt, 20 % had two or more), 42 % had previous amputations including 26.4 % of small amputations and 10.9 % of above‑knee amputations. The most common reason why patients in the treatment groups were not subject to the revascularization procedure was the technical impossibility of intervention (56.4 %), less common reason — the inappropriateness (33.6 %), which was evaluated on the Finnvasc scale.Conclusions. Critical lower limb ischemia is the most severe form of peripheral artery disease and is associated with a high risk of amputation of the affected limb and death. Three groups of patients, comparable in age, sex ratio, concomitant pathologies and clinical characteristics of the underlying disease, were selected for the prospective study. To compare the effectiveness of our treatment we selected a control group of patients retrospectively, by case histories, according to the inclusion criteria which were the same for all groups. This group was somewhat artificially created, as it consisted only of selected patients comparable to persons in treatment groups by age, comorbidities and underlying disease, as well as those patients whose treatment outcomes could be traced over a 3‑year follow‑up period.

Stem Cell Surgery and Growth Factors in Retinitis Pigmentosa Patients: Pilot Study after Literature Review

To evaluate whether grafting of autologous mesenchymal cells, adipose-derived stem cells, and platelet-rich plasma into the supracoroideal space by surgical treatment with the Limoli retinal restoration technique (LRRT) can exert a beneficial effect in retinitis pigmentosa (RP) patients. Twenty-one eyes underwent surgery and were divided based on retinal foveal thickness (FT) ≤ 190 or > 190 µm into group A-FT and group B-FT, respectively. The specific LRRT triad was grafted in a deep scleral pocket above the choroid of each eye. At 6-month follow-up, group B showed a non-significant improvement in residual close-up visus and sensitivity at microperimetry compared to group A. After an in-depth review of molecular biology studies concerning degenerative phenomena underlying the etiopathogenesis of retinitis pigmentosa (RP), it was concluded that further research is needed on tapeto-retinal degenerations, both from a clinical and molecular point of view, to obtain better functional results. In particular, it is necessary to increase the number of patients, extend observation timeframes, and treat subjects in the presence of still trophic retinal tissue to allow adequate biochemical and functional catering.

Genipin cross-linked decellularized scaffold for allogenic transplantation in situ

Objective To explored the bio-compatibility and cartilage regeneration of the rabbits genipin cross-linked decellularized scaffold, to provide experimental and theoretical support for the clinical application of genipin cross-linked decellularized scaffold. Methods Detergent-enzyme method was used to prepare decellularized tracheal scaffolds. Cellular content of native trachea and decellularized trachea were compared by 4′, 6-diamidino-2-phenylindole(DAPI) staining. Masson trichrome staining was used to compare the histological structure of the progenitor tube, decellularized trachea, and genipin cross-linked decellularized trachea. Nine adult New Zealand white rabbits were randomly divided into autologous tracheal transplantation group (negative control group), allogeneic tracheal transplantation group (positive control group), and genipin cross-linked decellularized tracheal transplantation group (experimental group). Autologous bone marrow mesenchymal stem cells were implanted on the surface of trachea in each group. The blood cells and type II collagen were detected to compare the inflammatory response and chondrocyte regeneration after tracheal orthotopic transplantation in the three groups. Results After DAPI staining and light microscope observation (×200), the cell content of the acellular 7-cycle trachea [(143.0 ± 71.1) cells/field] was significantly lower than that of the native trachea [(853.5 ± 149.6) cells/field], and the difference was statistically significant (P<0.001). Masson′s trichrome staining showed that the tissue structure of genipin cross-linked decellularized trachea was more complete. Blood cell analysis and type II collagen test results showed that genipin cross-linked decellularized trachea transplanted with bone marrow mesenchymal stem cells after transplantation in situ has little rejection and can be converted into chondrocytes by the action of related growth factors in vivo. Conclusions Genipin cross-linked decellularized tracheal scaffold combined with stem cell transplantation can successfully construct a tracheal in situ replacement model. This study provides a strong support for the research of tissue engineering trachea. Key words: Tissue engineering; Trachea; Transplantation; Genipin cross-linked; Mesenchymal stem cells

Treatment of complex recurrent fistula-in-ano by surgery combined to autologous bone marrow–derived mesenchymal stroma cells and platelet-rich plasma injection

We report a case of successful management of complex recurrent cryptoglandular fistula-in-ano by surgery combined with autologous bone marrow-derived mesenchymal stroma cells (MSCs) and platelet-rich plasma (PRP) injection. Clinical, radiological, and surgical data of the patient were reviewed, as well as the current literature on complex fistula-in-ano. A 37-year-old man with a recurrent cryptoglandular perianal fistula was addressed to our department. Inflammatory bowel disease was excluded by clinical history, endoscopy, and blood tests. Physical examination and MRI showed an anterior external orifice on the midline, 5cm from the anal verge, with an internal orifice on the same line. Surgery combined to injection of MSC-PRP solution was successfully performed. MSC-PRP solution was prepared while the patient was under general anesthesia: bone marrow MSCs were obtained by centrifugation of a tibial puncture specimen and PRP from a peripheral whole blood sample of the patient. There were no adverse events post-operatively. Clinical and MRI examination 4months after treatment confirmed the absence of perianal fistula. More than 2years after surgery, there has been no recurrence. Treatment of complex recurrent cryptoglandular fistula-in-ano by surgery combined to autologous bone marrow-derived MSCs and PRP injection seems safe in selected patients, allowing long-term healing. This procedure seems promising but further evaluation by clinical trials is warranted.

Long-term results of complex therapy in patients with nonreconstructible chronic lower limb ischemia

The aim — to improve the results of surgical treatment of patients with chronic critical lower limb ischemia by developing methods of autologous transplantation of multipotent stem mesenchymal stromal cells, administration of endothelial cells growth factors and therapeutic and diagnostic measures to preserve the lower limb or reduce the level of its amputation, reduce ischemia and the frequency of disability of such patients, improve their quality of life.Materials and methods. The study included 110 patients with a clinical picture of pain at rest, ischemic edema, and trophic changes (IIIA, IIIB, and IV stage of lower limb ischemia according to the classification of the European Association of Angiologists and Vascular Surgeons) caused by nonreconstructible occlusive and obliterating diseases of lower limb arteries. The patients were divided into three groups according to the method of treatment: in the 1st group (n = 48), revascularization osteotrepanation was performed as an operation of indirect revascularization, in the 2nd group (n = 21) a drug of endothelial cells growth factor was administered to stimulate their own angiogenesis, in the 3rd group (n = 41) subcutaneous fat was collected, autologous mesenchymal cells were cultivated and transplanted according to the developed technique. The control group of patients (n = 50) was selected retrospectively according to the case histories in accordance with the criteria of involvement and the results of treatment which could be followed up in a 3‑year observation period.Results and discussion. The safety of the lower limb in the 3rd group was statistically more significant than in the 1st and 2nd groups and remained at the level of 82 % after three years of observation. The smallest number of large amputations was in the 3rd group (2.3 % — above knee‑joint and 7.3 % — below, knee‑joint whereas in the control group the number was 18 % and 24 %, respectively).Conclusions. The level of high amputations directly affects the life expectancy of patients. Only treatment in the 3rd group reduced the risk of large amputation compared with the control group (odds ratio 2.752 (95 % confidence interval — 1.12 — 6.733)). Only treatment in the 3rd group reduced the risk of amputation compared with other types of treatment (odds ratio 0.358 (95 % confidence interval 1.111 — 1.158).

Periosteum-derived mesenchymal progenitor cells in engineered implants promote fracture healing in a critical-size defect rat model.

An attractive alternative to bone autografts is the use of autologous mesenchymal progenitor cells (MSCs) in combination with biomaterials. We compared the therapeutic potential of different sources of mesenchymal stem cells in combination with biomaterials in a bone nonunion model. A critical-size defect was created in Sprague-Dawley rats. Animals were divided into six groups, depending on the treatment to be applied: bone defect was left empty (CTL); treated with live bone allograft (LBA); hrBMP-2 in collagen scaffold (CSBMP2 ); acellular polycaprolactone scaffold (PCL group); PCL scaffold containing periosteum-derived MSCs (PCLPMSCs ) and PCL containing bone marrow-derived MSCs (PCLBMSCs ). To facilitate cell tracking, both MSCs and bone graft were isolated from green fluorescent protein (GFP)-transgenic rats. CTL group did not show any signs of healing during the radiological follow-up (n=6). In the LBA group, all the animals showed bone bridging (n=6) whereas in the CSBMP2 group, four out of six animals demonstrated healing. In PCL and PCLPMSCs groups, a reduced number of animals showed radiological healing, whereas no healing was detected in the PCLBMSCs group. Using microcomputed tomography, the bone volume filling the defect was quantified, showing significant new bone formation in the LBA, CSBMP2 , and PCLPMSCs groups when compared with the CTL group. At 10weeks, GFP positive cells were detected only in the LBA group and restricted to the outer cortical bone in close contact with the periosteum. Tracking of cellular implants demonstrated significant survival of the PMSCs when compared with BMSCs. In conclusion, PMSCs improve bone regeneration being suitable for mimetic autograft design.

The use of adipose stem cells and the tubulization technique on regeneration of the facial nerve in wistar rats after transection with loss of substance

Lesions in peripheral nerves are quite frequent and can cause serious damage to the patient. When the facial nerve is affected, patients may present both physical functional and emotional changes, causing devastating consequences to these people. Despite all the evolution of reparative microsurgery and tissue engineering, the results are still not totally satisfactory and further studies are still needed. This work consists of an experimental study in which transection with loss of substance on the facial nerve on wistar rats and subsequent treatment by the tubulization technique in comparison to the use of the autologous graft were carried out. 72 male wistar rats were recruited and divided into groups A, B and C, in which a 6 mm defect was created in the facial nerve on the right side. The treatment of the nerve in group A was performed by autologous graft (golden standard). In group B, the technique of tubulization using an absorbable polymer (PLGA PCL) sutured in both proximal and distal stumps of the nerve. In group C, the same absorbable tube with the addition of autologous mesenchymal cells extracted from the adipose tissue of the animals was used. The assessment was performed at three different moments, (30, 60 and 90 days) in which histological sections stained with HE and Masson's trichrome were performed. The results were obtained in relation to fibrosis, granulomatous infiltrate and nerve regeneration found in histological sections. At 30 days, all animals had the same percentage of nerve regeneration. At 60 days, the tubulization presented better results than the gold standard. At 90 days, it was observed that the use of stem cells did not improve the results in relation to nerve regeneration. At the end of 90 days, it was observed that all the animals showed high percentages of nerve regeneration suggesting that if the evaluation were performed in larger periods, the regeneration indexes would be even higher. It is of great clinical relevance that more studies should be performed in the area of peripheral nerve regeneration, especially in periods greater than 90 days. It is also perceived that the association of neurotrophic factors with tissue engineering seems to be a very promising path.

Safety of autologous bone marrow mesenchymal stem cells transplantation in proliferative diabetic retinopathy patients

Objective To investigate the safety of autologous bone marrow mesenchymal stem cells (ABMSCs) transplantation into the subretinal space for the treatment of proliferative diabetic retinopathy (PDR). Methods The clinical data of four PDR patients (four eyes) who received ABMSCs transplantation into the subretinal space were collected in Army Medical University, Southwest Eye Hospital from March 2014 to December 2015, including 3 males and 1 female; the average age was 55 years old; the average course of diabetes was 10 years, and the blood glucoses were all well controlled before treatment.All the patients underwent conventional ophthalmologic examination, and visual acuity, slit lamp microscope, color fundus photography, fluorescein angiography (FFA) and optical coherence tomography (OCT) examination were performed at 1 week, 1 month, 3 months, 6 months, 9 months and 12 months after surgery.This study protocol was approved by Ethic Committee of Army Medical University, Southwest Eye Hospital (No.2013-34). Results Four patients diagnosed as PDR were enrolled in this study.All patients were performed ABMSC transplantation, and no one felt discomfort after treatment.FFA and OCT showed that the transplanted cells were present in the subretinal space until 1 month after transplantation.The macular edema of one patient diagnosed as macular edema preoperatively was relived gradually after transplantation, and the effects lasted 3 months after transplantation.The preoperative best corrected visual acuity (BCVA) of the two patients were improved from hand movement and finger counting to 20/20 (84 ETDRS) and 20/200 (38 ETDRS) after transplantation, respectively, and the visual acuities of the other two eyes were both stable.All patients underwent panretinal photocoagulatio 3 months after transplantation, and the follow-up treatment complied with the routine of post-vitrectomy for DR, no complications occurred during the follow-up period. Conclusions Subretinal transplantation of ABMSCs for PDR is safe.The transplanted cells show local anti-inflammatory effect, and no effect on cell proliferation or circulatory improvement are observed. Key words: Diabetic retinopathy; Autologous bone marrow mesenchymal stem cell; Subretinal transplantation

Клинико-морфологическая характеристика больных до и после трансплантации аутологичныхмезенхимальных стволовых клеток костного мозга при Системной склеродермии

Клинико-морфологическая характеристика больных до и после трансплантации аутологичныхмезенхимальных стволовых клеток костного мозга при Системной склеродермии

Клинико-морфологическая характеристика больных до и после трансплантации аутологичныхмезенхимальных стволовых клеток костного мозга при Системной склеродермии

Клинико-морфологическая характеристика больных до и после трансплантации аутологичныхмезенхимальных стволовых клеток костного мозга при Системной склеродермии

Bone Marrow Stromal Cells Promote Innervation of Bioengineered Teeth

Transplantation of bone marrow mesenchymal stem cells (BMDCs) into a denervated side of the spinal cord was reported to be a useful option for axonal regeneration. The innervation of teeth is essential for their function and protection but does not occur spontaneously after injury. Cultured reassociations between dissociated embryonic dental mesenchymal and epithelial cells and implantation lead to a vascularized tooth organ regeneration. However, when reassociations were coimplanted with a trigeminal ganglion (TG), innervation did not occur. On the other hand, reassociations between mixed embryonic dental mesenchymal cells and bone marrow–derived cells isolated from green fluorescent protein (GFP) transgenic mice (BMDCs-GFP) (50/50) with an intact and competent dental epithelium (ED14) were innervated. In the present study, we verified the stemness of isolated BMDCs, confirmed their potential role in the innervation of bioengineered teeth, and analyzed the mechanisms by which this innervation can occur. For that purpose, reassociations between mixed embryonic dental mesenchymal cells and BMDCs-GFP with an intact and competent dental epithelium were cultured and coimplanted subcutaneously with a TG for 2 wk in ICR mice. Axons entered the dental pulp and reached the odontoblast layer. BMDCs-GFP were detected at the base of the tooth, with some being present in the pulp associated with the axons. Thus, while having a very limited contribution in tooth formation, they promoted the innervation of the bioengineered teeth. Using quantitative reverse transcription polymerase chain reaction and immunostainings, BMDCs were shown to promote innervation by 2 mechanisms: 1) via immunomodulation by reducing the number of T lymphocytes (CD3+, CD25+) in the implants and 2) by expressing neurotrophic factors such as NGF, BDNF, and NT3 for axonal growth. This strategy using autologous mesenchymal cells coming from bone marrow could be used to innervate bioengineered teeth without treatment with an immunosuppressor such as cyclosporine A (CsA), thus avoiding multiple side effects.