Mesenchymal Cell Transplantation Research Articles

Evaluation of the Safety and Efficacy of Repeated Mesenchymal Stem Cell Transplantations in ALS Patients by Investigating Patients' Specific Immunological and Biochemical Biomarkers.

Amyotrophic lateral sclerosis (ALS) is an incurable disease. There are vigorous attempts to develop treatments to reduce the effects of this disease, and among these treatments is the transplantation of stem cells. This study aimed to retrospectively evaluate a mesenchymal stem cell (MSC) therapy cohort as a promising novel treatment modality by estimating some additional new parameters, such as immunological and biochemical factors. This study was designed as an open-label, one-arm cohort retrospective study to evaluate potential diagnostic biomarkers of repeated infusions of autologous-bone marrow-derived mesenchymal stem cells (BM-MSCs) in 15 confirmed patients with ALS, administered at a dose of 1 × 106 cells/kg BW with a one-month interval, in equal amounts in both an intravenous (IV) and intrathecal (IT) capacity simultaneously, via various biochemical (iron (Fe), ferritin, total-iron-binding capacity (TIBC), transferrin, and creatine kinase (CK)) and immunological parameters (tumor necrosis factor-alpha (TNF-α), neurofilament light chain (NFL), and glial-cell-derived neurotrophic factor (GDNF) levels, evaluated during the three-month follow-up period in serum and cerebrospinal fluid (CSF). Our study indicated that, in the case of immunological biomarkers, TNF-α levels in the CSF showed a significant decrease at month three after transplantation compared with levels at month zero, and the p-value was p < 0.01. No statistically significant changes were observed for other immunological as well as biochemical parameters and a p-value of p > 0.05. These results can indicate the potential benefit of stem cell transfusion in patients with ALS and suggest some diagnostic biomarkers. Several studies are required to approve these results.

Mesenchymal stem cells suppressed skin and lung inflammation and fibrosis in topoisomerase I-induced systemic sclerosis associated with lung disease mouse model.

Systemic sclerosis associated with lung interstitial lung disease (SSc-ILD) is the most common cause of death among patients with SSc. Mesenchymal stem cell (MSCs) transplantations had been treated by SSc patients that showed in the previous case report. The therapeutic mechanisms and effects of MSCs on SSc-ILD are still obscure. In this study, we investigated the therapeutic effects and mechanisms of treatment of BM-MSC derived from C57BL/6 on the topoisomerase I (TOPO I) induced SSc-ILD-like mice model. The mice were immunized with a mixture of recombinant human TOPO I in PBS solution (500 U/mL) and completed Freund's adjuvant [CFA; 1:1 (volume/volume)] twice per week for 9weeks. On week 10, the mice were sacrificed to analyze the related pathological parameters. Lung and skin pathologies were analyzed using histochemical staining. CD4 T-helper (TH) cell differentiation in lung and skin-draining lymph nodes was detected using flow cytometry. Our results revealed that allogeneic and syngeneic MSCs exhibited similar repressive effects on TOPO I-induced IgG1 and IgG2a in the SSc group. After intravascular (IV) treatment with syngeneic or allogeneic MSCs, the dermal thickness and fibrosis dramatically condensed and significantly reduced airway hyperresponsiveness. These findings showed that both allogeneic and syngeneic MSCs have therapeutic potential for SSc-ILD.

Injectable cultured bone marrow derived mesenchymal cells vs chondrocytes in the treatment of chondral defects of the knee – RCT with 6 years follow-up

Articular cartilage has unique biological and biomechanical characteristics. Damage to this tissue fails to heal spontaneously, leading to progressive arthritis. Cartilage repair techniques have been looked forward to in the treatment of significant cartilage injuries. Cell-based regenerative techniques like the two-staged cultured chondrocytes and single-stage mesenchymal cell transplantation have been tried with varying results and limitations. We study the outcomes of cultured bone marrow derived MSCs in the treatment of articular cartilage defects of the knee in comparison to autologous cultured chondrocyte implantation (ACI). Both cultured MSC and ACI treatment methods resulted in significant improvements in patient reported outcome measures (PROMs). There was no difference in the PROMs, MOCART scores, T2∗ mapping and dGEMRIC values between the groups. Use of cultured MSCs leads to good clinical outcomes similar to ACI and represents a promising treatment to restore the articular cartilage in the knee.

The Effects of the Combination of Mesenchymal Stromal Cells and Nanofiber-Hydrogel Composite on Repair of the Contused Spinal Cord.

A bone marrow-derived mesenchymal stromal cell (MSC) transplant and a bioengineered nanofiber-hydrogel composite (NHC) have been shown to stimulate nervous tissue repair in the contused spinal cord in rodent models. Here, these two modalities were combined to assess their repair effects in the contused spinal cord in adult rats. Cohorts of contused rats were treated with MSC in NHC (MSC-NHC), MSC in phosphate-buffered saline (MSC-PBS), NHC, or PBS injected into the contusion site at 3 days post-injury. One week after injury, there were significantly fewer CD68+ cells in the contusion with MSC-NHC and NHC, but not MSC-PBS. The reduction in CD86+ cells in the injury site with MSC-NHC was mainly attributed to NHC. One and eight weeks after injury, we found a greater CD206+/CD86+ cell ratio with MSC-NHC or NHC, but not MSC-PBS, indicating a shift from a pro-inflammatory towards an anti-inflammatory milieu in the injury site. Eight weeks after injury, the injury size was significantly reduced with MSC-NHC, NHC, and MSC-PBS. At this time, astrocyte, and axon presence in the injury site was greater with MSC-NHC compared with MSC-PBS. We did not find a significant effect of NHC on MSC transplant survival, and hind limb function was similar across all groups. However, we did find fewer macrophages at 1 week post-injury, more macrophages polarized towards a pro-regenerative phenotype at 1 and 8 weeks after injury, and reduced injury volume, more astrocytes, and more axons at 8 weeks after injury in rats with MSC-NHC and NHC alone compared with MSC-PBS; these findings were especially significant between rats with MSC-NHC and MSC-PBS. The data support further study in the use of an NHC-MSC combination transplant in the contused spinal cord.

Safety of Intravenous Autologous Bone Marrow-Derived Mesenchymal Cell Transplantation in 5 Patients With Reduced Left Ventricular Ejection Fraction.

Background: Although intracardiac injection or intracoronary delivery of mesenchymal stem cells (MSCs) has been reported, there have been few studies on the intravenous injection of MSCs, particularly in Japan.Methods and Results: Five patients with left ventricular ejection fraction (LVEF) ≤45% received 1.0×108 MSCs intravenously. The procedure did not induce significant changes in vital signs. One patient had an elevated body temperature after 1 day, but recovered spontaneously. Laboratory tests remained normal for 1 month after cell delivery. Computed tomography was performed after 1–2 years, and there was no evidence of malignancy.Conclusions: In this pilot study of patients with reduced LVEF, intravenous MSC delivery had no adverse effects.

Potential therapeutic uses of intraoral mesenchymal stem cells in other tissues of the body: A review.

BackgroundOver the last few years, there has been a great advance in regenerative medicine, with various studies that have observed the ability to repair or regenerate dysfunctional tissues with the patient’s own cells, such as with mesenchymal cells. In this area, mesenchymal stem cells (MSCs) from the oral cavity have attracted attention because of their easy access and multiple cellular differentiations. Multiple studies have shown the various clinical applications at the intraoral level, especially at the level of bone regeneration, but the potential applications of oral MSC at a systemic level have been scarcely described. Objective: The objective of this review was to describe the potential therapeutic uses of intraoral MSCs in other tissues of the organism. Material and MethodsA review of the literature between 2000 and 2019. Only included those studies done on animals or humans. ResultsTwenty five articles were selected, all performed on animals. The donor site most used were the temporary teeth exfoliated from humans, representing 56% of the total articles, followed by the dental pulp with 28% of the total articles included. Transplantation of intraoral mesenchymal cells in animals with neural tissue illness was the most studied therapy. ConclusionsAlthough obtaining MSC of intraoral origin has proven to be a good alternative in regenerative medicine, achieving therapeutic uses in bone tissue, nervous tissue, liver tissue, skin tissue, ocular tissue, reperfusion of tissues and in autoimmune diseases, there is a lack of clinical studies that allow its safe use in humans. Key words:Mesenchymal stem cells, stem cell transplantation, regenerative medicine, dental component.

Idiopathic pulmonary hemosiderosis: a review of the treatments used during the past 30years and future directions.

This paper reviews the literature on the treatment modalities for idiopathic pulmonary hemosiderosis (IPH) used over the past 30years, attempting to define treatment options that appear to be efficacious and safe, and in addition presents a treatment algorithm. IPH is an uncommon etiology of diffuse alveolar hemorrhage. IPH is a rare disease in adults and often associated with a significant temporal delay in diagnosis. Patients present with hemoptysis, radiographic chest abnormalities, and iron deficiency anemia. Although several pathogenetic hypotheses have been proposed, IPH appears to be an immunologic disease, possibly with a genetic component. Corticosteroid therapy represents the first line of treatment, including liposome-incorporated dexamethasone palmitate (liposteroid). Additional immunomodulatory/immunosuppressive medications have been used with varying success, especially in the setting of steroid-refractory disease. Cyclophosphamide, azathioprine, hydroxychloroquine, mycophenolate mofetil, and mesenchymal cell transplantation have been attempted to improve outcome and reduce side effects. Controlled studies are needed to assess the optimal combination of medications, which are effective to control the disease.

Transplantation of mesenchymal stem cells preserves podocyte homeostasis through modulation of parietal epithelial cell activation in adriamycin-induced mouse kidney injury model.

To determine the role of the transplantation of bone marrow-derived mesenchymal stem cells (MSCs) in podocyte renewal, we studied BALB/C mice with or without adriamycin-induced acute kidney injury. MSCs were transplanted ectopically under the capsule of the left kidney or into the peritoneal cavity after the onset of kidney injury to test testing their local or systemic paracrine effects, respectively. Adriamycin produced increases in urine protein: creatinine ratios, blood urea nitrogen, and blood pressure, which improved after both renal subcapsular and intraperitoneal MSCs transplants. The histological changes of adriamycin kidney changes regressed in both kidneys and in only the ipsilateral kidney after intraperitoneal or renal subcapsular transplants indicating that the benefits of transplanted MSCs were related to the extent of paracrine factor distribution. Analysis of kidney tissues for p57-positive parietal epithelial cells (PECs) showed that MSC transplants restored adriamycin-induced decreases in the abundance of these cells to normal levels, although after renal subcapsular transplants these changes did not extend to contralateral kidneys. Moreover, adriamycin caused inflammatory activation of PECs with coexpression of CD44 and phospho-ERK, which was normalized in both or only ipsilateral kidneys depending on whether MSCs were transplanted in the peritoneal cavity or subcapsular space, respectively.

Autologous Bone Marrow Mesenchymal Cell Transplantation in the Treatment of Freiberg's Disease: Case Report

Background: Osteonecrosis of the metatarsal head, classically known as Freiberg's disease, presents as aseptic necrosis of the epiphysis of undefined etiology, although it may be associated with trauma, prolonged corticotherapy or systemic diseases such as sickle cell disease. There are several techniques described for the treatment of osteonecrosis secondary to sickle cell disease. Methods: Case report and the surgical description of the use of autologous bone marrow mesenchymal stem cell transplantation for the treatment of osteonecrosis of the 2nd metatarsal head. Results: Nuclear magnetic resonance imaging performed after one year of follow-up showed a regression of the findings found in the preoperative evaluation. Conclusion: This is the first time that the technique of autologous bone marrow mesenchymal stem cell implantation has been employed for the treatment of osteonecrosis of the metatarsal head. The case reported shows that the technique is safe and brings good results after conservative treatment failure.

An experimental model of acute cerebral ischemia with transplantation of mesenchymal stem cells into the carotid artery

Transplantation of mesenchymal cells is a perspective paradigm for treatment of stroke. However, after intravenous injection most infusion cells get in filter organs, such as lungs. Experimental model of cerebral ischemia with transplantation of mesenchymal cells in a carotid was developed. Experiments on 15 animals conducted on by the females of crawls in age 12-36 weeks weighing 4-6 kg. The general and internal carotid arteries were projected under local and intravenous anesthesia. General carotid artery was ligated and mesenchymal stem cells were injected. The population of the cages abstracted from a brain was folded on 96% from cages positive for by the markers of mesenchymal cells, and less than on 2% from hematopoietic cells and cells of endothelia, positive after corresponding markers. In 6 hour after introduction to the right internal carotid of mesenchymal cells, the histological encephaloscopy of crawls was executed for visualization of GFP-positive cages mark also the marker of lipophil of PKH26. In 6 hour after transplantation of cage distributed at a right hemisphere in area of bark and basale kernels (in the zone of blood supply of right internal carotid) and visualized along the midwall of cerebral vessels both in the zone of heart attack of brain and for peripheries. Experimental confinnation of that therapeutic activity of mesenchymal cells arises up during their system transplantation and delivery on an artery that supplies with blood the zone of ischemic damage of brain is first got. Keywords: carotid arteries, mesenchymal stem cells, ischemic stroke.

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.

Injectable hydrogel combined with nucleus pulposus-derived mesenchymal stem cell for the treatment of degenerative intervertebral disc in rats

Objective To investigate the effect of injectable hydrogel combined with nucleus pulposus-derived mesenchymal stem cell (NPMSC) transplantation on the degenerative intervertebral disc in rats. Methods NPMSC was isolated from the caudal spinal nucleus pulposus of Sprague-Dawley rats and identification of mesenchymal stem cells was determined. After 2 weeks of intervertebral disc degeneration model was established by annulus fibrosus puncture, the groups were divided: control group (untreated), treatment group (injection of hydrogel loaded NPMSC into the intervertebral disc), phosphate buffer solution (PBS) group (injection of the same amount of PBS), hydrogel group (injection of the same amount of hydrogel), cell group (injection of the same amount of NPMSC suspension). At 4 and 8 weeks after intervention, relative disc height index (DHI%) and magnetic resonance imaging (MRI) index were detected. At 8 weeks after intervention, histological score, the expression of collagen type Ⅱ and aggrecan was detected by immunofluorescence and reverse transcription-polymerase chain reaction. One-way ANOVA was used for comparison between groups, and LSD-t test was used for pairwise comparison. Results At 8 weeks after intervention, for the DHI% and MRI index, the experiment group [(87.80±3.40)%, 3 313.84±227.80] was higher than cell group [(79.65±2.33)%, 3 006.83±147.92] (t=3.411, 3.127, P<0.05); cell group was higher than hydrogel group [(69.49±4.79)%, 1 288.00±202.09] (t=4.252, 17.504, P<0.05); for histological score, the cell group (9.00±1.29) was higher than experiment group (7.16±0.68) (t=3.550, P<0.05), the hydrogel group (12.50±0.95) was higher than cell group (t=6, 753, P<0.05); for the protein express of collagen type Ⅱ and aggrecan, the cell group (0.30±0.02, 0.58±0.03) was less than experiment group (0.38±0.03, 0.69±0.05) (t=-4.000, -3, 267, P<0.05), the hydrogel group (0.14±0.03, 0.14±0.04) was less than cell group (t=-8.000, -13.307, P<0.05). Conclusion Injectable hydrogel combined with NPMSC transplantation can delay the level of disc degeneration in animal model and promote the repair and regeneration of degenerative disc. Key words: Hydrogel; Nucleus pulposus-derived mesenchymal stem cell; Intervertebral disc degeneration; Cell transplantation

The effects of bone marrow mesenchymal stem cells on the expression of nerve growth factor and caspase-3 after cardiac arrest in the rats

Objective To investigate the effects of bone marrow mesenchymal stem cells (BMSCs) transplantation on the expression of nerve growth factor (NGF) and Caspase-3 in rat hippocampus after cardiac arrest (CA). Methods Sprague-Dawley (SD) rats were randomly divided into 3 groups: sham group (n=6), CA group (n=6), and BMSCs group (n=6). CPR was performed on the groups after the induction of asphyxial cardiac arrest. Animals in the BMSCs group or the CA group were respectively injected with a dose of 1×106 BMSCs in 0.5 mL phosphate buffer solution (PBS) or 0.5 mL PBS alone via the vena caudalis 1 h after successful resuscitation. The neurological status after restoration of spontaneous circulation (ROSC) were assessed by modified neurological severity scores (mNSS); serum levels of S100B were assayed, and the expression of NGF and Caspase-3 in hippocampus was detected by immunohistochemistry. Results Compared with the CA group, mNSS and S100B levels were lower in the BMSCs group on the 7th day after ROSC [(0.9±0.3) vs (4.5±0.6), (90.12±4.62) pg/mL vs (182.30±2.58) pg/mL, both P<0.05] with higher expression of NGF and lower expression of Caspase-3 [(11.391±1.297) vs (7.744±1.334), (6.256±1.036) vs (8.506±1.742), both P< 0.05]. Conclusions BMSCs transplantation might improve rat's neurological functions after cardiac arrest, which may be related to up-regulation of NGF expression and down-regulation of Caspase-3 expression. Key words: Bone marrow mesenchymal stem cells; Cardiac arrest; Global cerebral ischemic injury; Cerebral resuscitation; Transplantation; Nerve growth factor; Caspase-3

Pilot experimental study on amniotic epithelial mesenchymal cell transplantation in natural occurring tendinopathy in horses. Ultrasonographic and histological comparison

amnion-derived stem cells are considered a promising alternative source for tendon tissue regeneration. aims of this paper were to illustrate the ultrasound and histological outcomes following the treatment of acute and chronic superficial digital flexor tendon spontaneous lesions in horses with ovine amniotic epithelial cells xenotransplantation. six adult horses suffering from unilateral acute (4 cases) and chronic (2 cases) tendinopathy (clinical and ultrasound diagnosis) were enrolled. At baseline, ovine amniotic epithelial cells were grafted, in sterile conditions and under ultra-sound control, into the most damaged area. Ultra-sound controls were performed at 30, 60, 90, 120, 150 and 180 days after cells implantation; after horse euthanasia (180 days) tendon samples were collected and submitted to histological examination (cellularity, extracellular matrix fiber organization, blood vessels). at baseline, in the acute cases, the ultra-sound exam showed a focal, dis-homogeneous, hypo-echoic area into the superficial digital flexor tendon, with loss of the normal fibrillar pattern, while in the chronic cases the damaged tendon area appeared thickened and completely hyper-echoic. At the final follow-up tendon echotexture was more regular, the cross-sectional area similar to the contra-lateral limb, and the collagen fibers were oriented in parallel to the longitudinal axis of the tendon both in the acute and chronic cases, suggesting a positive healing response. These findings were supported by the histological analyses which showed an almost complete restoration of normal tendon architecture with an optimal alignment of tendon fibers. the present pilot study supports the hypothesis that amniotic epithelial cells are provided of an excellent healing potential and shows a very good correlation between the ultrasound findings and the histologic features.

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

Tenogenic Properties of Mesenchymal Progenitor Cells Are Compromised in an Inflammatory Environment

Transplantation of multipotent mesenchymal progenitor cells is a valuable option for treating tendon disease. Tenogenic differentiation leading to cell replacement and subsequent matrix modulation may contribute to the regenerative effects of these cells, but it is unclear whether this occurs in the inflammatory environment of acute tendon disease. Equine adipose-derived stromal cells (ASC) were cultured as monolayers or on decellularized tendon scaffolds in static or dynamic conditions, the latter represented by cyclic stretching. The impact of different inflammatory conditions, as represented by supplementation with interleukin-1β and/or tumor necrosis factor-α or by co-culture with allogeneic peripheral blood leukocytes, on ASC functional properties was investigated. High cytokine concentrations increased ASC proliferation and osteogenic differentiation, but decreased chondrogenic differentiation and ASC viability in scaffold culture, as well as tendon scaffold repopulation, and strongly influenced musculoskeletal gene expression. Effects regarding the latter differed between the monolayer and scaffold cultures. Leukocytes rather decreased ASC proliferation, but had similar effects on viability and musculoskeletal gene expression. This included decreased expression of the tenogenic transcription factor scleraxis by an inflammatory environment throughout culture conditions. The data demonstrate that ASC tenogenic properties are compromised in an inflammatory environment, with relevance to their possible mechanisms of action in acute tendon disease.

Cortical AAV-CNTF Gene Therapy Combined with Intraspinal Mesenchymal Precursor Cell Transplantation Promotes Functional and Morphological Outcomes after Spinal Cord Injury in Adult Rats.

Ciliary neurotrophic factor (CNTF) promotes survival and enhances long-distance regeneration of injured axons in parts of the adult CNS. Here we tested whether CNTF gene therapy targeting corticospinal neurons (CSN) in motor-related regions of the cerebral cortex promotes plasticity and regrowth of axons projecting into the female adult F344 rat spinal cord after moderate thoracic (T10) contusion injury (SCI). Cortical neurons were transduced with a bicistronic adeno-associated viral vector (AAV1) expressing a secretory form of CNTF coupled to mCHERRY (AAV-CNTFmCherry) or with control AAV only (AAV-GFP) two weeks prior to SCI. In some animals, viable or nonviable F344 rat mesenchymal precursor cells (rMPCs) were injected into the lesion site two weeks after SCI to modulate the inhibitory environment. Treatment with AAV-CNTFmCherry, as well as with AAV-CNTFmCherry combined with rMPCs, yielded functional improvements over AAV-GFP alone, as assessed by open-field and Ladderwalk analyses. Cyst size was significantly reduced in the AAV-CNTFmCherry plus viable rMPC treatment group. Cortical injections of biotinylated dextran amine (BDA) revealed more BDA-stained axons rostral and alongside cysts in the AAV-CNTFmCherry versus AAV-GFP groups. After AAV-CNTFmCherry treatments, many sprouting mCherry-immunopositive axons were seen rostral to the SCI, and axons were also occasionally found caudal to the injury site. These data suggest that CNTF has the potential to enhance corticospinal repair by transducing parent CNS populations.

Efficacy and long-term longitudinal follow-up of bone marrow mesenchymal cell transplantation therapy in a diabetic patient with recurrent lower limb bullosis diabeticorum

Bullosis diabeticorum is a rare presentation of cutaneous manifestation most commonly affecting the lower limbs in patients with diabetes. The appearance, often as insidious as its resolution, is characterized by tense blisters on the skin surfaces of the lower limbs and the feet. The cause still remains unclear, but it may relate to microangiopathy and neuropathy. In this report, we present a case of a 64-year-old male with multiple episodes of blistering in the left lateral lower limb after a traumatic fall who was subsequently diagnosed with type 2 diabetes mellitus. The patient had a history of poorly controlled blood glucose and subsequently developed vasculopathy and peripheral neuropathy. Despite appropriate glycemic control and antibiotics therapy, the patient developed recurrent bullosis diabeticorum on five separate occasions during a 2-year span from 2005 to 2007. Building on our success with ischemic diabetic foot, we used bone marrow mesenchymal stem cell (BMMSC) transplantation therapy for bullosis diabeticorum. After a 9-month treatment, this patient developed another episode of cellulitis in the same lower limb which was successfully treated with antibacterial therapy. It is interesting that the patient reported no recurrence in the next 10-year follow-up span. This study demonstrates that bullosis diabeticorum could appear even before the onset of diabetes, and vascular insufficiency predisposes to the occurrence of bullosis diabeticorum. Our findings suggest that autologous BMMSC transplantation therapy may be an effective measure for recurrent bullosis diabeticorum; however, this will require further investigation to be conclusive. Early identification of diabetes and its complications and appropriate treatment may improve clinical outcomes and prevent lower limb amputation.Trial registration: ClinicalTrials.gov Identifier: NCT00955669. Registered on August 10, 2009.

Effects of bone marrow mesenchymal stem cell transplantation on retinal neovascularization in neonatal rats with oxygen-induced retinopathy

To explore the effects of rat bone mesenchymal stem cell (BMSC) transplantation on retinal neovascularization, and to observe the changes of hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factors (VEGF) in rats with oxygen-induced retinopathy (OIR). Seventy-two seven-day-old Sprague-Dawley rats were randomly divided into three groups: normal control (CON), model (OIR) and BMSC transplantation. In the BMSC transplantation group, BMSCs were transplanted 5 days after oxygen conditioning. The phosphate buffered saline of the same volume was injected in the CON and OIR groups. The OIR model was prerpared according to the classic hyperoxygen method. At seven days after transplantation, retinal neovascularization was examined by retinal flat-mount staining and hematoxylin eosin (HE) staining. The expression of HIF-1α and VEGF proteins was examined by immunohistochemistry staining and Western blot analysis. The retinal flat-mount staining results showed that the vessels were well organized in the CON group, but the vessels were irregularly organized, and lots of nonperfusion areas were observed in the OIR group. The large vessels were a bit circuitous, the retinal vessels were relatively organized, and less nonperfusion areas were noted in the BMSC transplantation group. The HE staining results showed that many neovessels and preretinal neovascular (pre-RNC) cells were observed on the internal limiting membrane in the OIR group. There were less pre-RNC cells in the BMSC transplantation group compared with the OIR group (P<0.01). The immunohistochemistry analysis showed that more HIF-1α+ and VEGF+ cells were observed in the OIR group compared with the CON group, and less HIF-1α+ and VEGF+ cells were observed in the BMSC transplantation group compared with OIR group (P<0.05). The Western blot analysis showed the expression of HIF-1α and VEGF proteins in the OIR group was significantly higher than that in the CON group. The expression of HIF-1α and VEGF proteins in the BMSC transplantation group was lower than that in the OIR group (P<0.01). BMSC transplantation therapy could alleviate retinal neovascularization in OIR rats, and its mechanisms might be associated with the inhibition of the expression of HIF-1α and VEGF proteins.

Evaluating the biological risk of functionalized multiwalled carbon nanotubes and functionalized oxygen-doped multiwalled carbon nanotubes as possible toxic, carcinogenic, and embryotoxic agents.

Carbon nanotubes (CNTs) have been a focus of attention due to their possible applications in medicine, by serving as scaffolds for cell growth and proliferation and improving mesenchymal cell transplantation and engraftment. The emphasis on the benefits of CNTs has been offset by the ample debate on the safety of nanotechnologies. In this study, we determine whether functionalized multiwalled CNTs (fMWCNTs) and functionalized oxygen-doped multiwalled CNTs (fCOxs) have toxic effects on rat mesenchymal stem cells (MSCs) in vitro by analyzing morphology and cell proliferation and, using in vivo models, whether they are able to transform MSCs in cancer cells or induce embryotoxicity. Our results demonstrate that there are statistically significant differences in cell proliferation and the cell cycle of MSCs in culture. We identified dramatic changes in cells that were treated with fMWCNTs. Our evaluation of the transformation to cancer cells and cytotoxicity process showed little effect. However, we found a severe embryotoxicity in chicken embryos that were treated with fMWCNTs, while fCOxs seem to exert cardioembryotoxicity and a discrete teratogenicity. Furthermore, it seems that the time of contact plays an important role during cell transformation and embryotoxicity. A single contact with fMWCNTs is not sufficient to transform cells in a short time; an exposure of fMWCNTs for 2 weeks led to cell transformation risk and cardioembryotoxicity effects.