Recurrent anal fistulae:Limited surgery supported by stem cells

AIM:To study the results of stem-cell therapy under a Compassionate-use Program for patients with recurrent anal fistulae.METHODS:Under controlled circumstances,and approved by European and Spanish laws,a Compassionate-use Program allows the use of stem-cell therapy for patients with very complex anal fistulae.Candidates had previously undergone multiple surgical interventions that had failed to resolve the fistulae,and presented symptomatic recurrence.The intervention consisted of limited surgery(with closure of the internal opening),followed by local implant of stem cells in the fistula-tract wall.Autologous expanded adipose-derived stem cells were the main cell type selected for implant.The first evaluation was performed on the 8th postoperative week;outcome was classified as response or partial response.Evaluation one year after the intervention confirmed if complete healing of the fistula was achieved.RESULTS:Ten patients(8 male)with highly recurrent and complex fistulae were treated(mean age:49years,range:28-76 years).Seven cases were nonCrohn’s fistulae,and three were Crohn’s-associated fistulae.Previous surgical attempts ranged from 3to 12.Two patients presented with preoperative incontinence(Wexner scores of 12 and 13 points).After the intervention,six patients showed clinical response on the 8th postoperative week,with a complete cessation of suppuration from the fistula.Three patients presented a partial response,with an evident decrease in suppuration.A year later,six patients(60%)remained healed,with complete reepithelization of the external opening.Postoperative Wexner Scores were 0 in six cases.The two patients with previous incontinence improved their scores from12 to 8 points and from 13 to 5 points.No adverse reactions or complications related to stem-cell therapy were reported during the study period.CONCLUSION:Stem cells are safe and useful for treating anal fistulae.Healing can be achieved in severe cases,sparing fecal incontinence risk,and improving previous scoring.

Immunophenotyping of hematopoietic progenitor cells: Comparison between cord blood and adult mobilized blood grafts

AIM:To study the immunophenotype of hematopoietic progenitor cells from cord blood (CB) grafts (n = 39) in comparison with adult apheresis grafts (AG, n = 229) and pre-apheresis peripheral blood (PAPB) samples (n = 908) using flow cytometry analysis.METHODS: First, we performed a qualitative analysis of CD34+ cell sub-populations in both CB and PAPB grafts using the standardized ISHAGE protocol and a wide panel of 20 monoclonal antibodies. Next, we stud-ied some parameters, such as the age of mothers and the weight of newborns, which can influence the qual-ity and the quantity of CD34+ cells from CB. RESULTS: We found that the percentage of apoptotic cells was high in CB in comparison to PAPB (PAPB: 4.6% ± 2.6% vs CB: 53.4% ± 5.2%, P < 0.001). In CB, the weight of newborn and the age of the mother have the influence on CD34+ cells. The follow-up of Ag CD133in the ISHAGE double platform protocol in association with CD45, CD34 and the 7’AAD shows an equal rate between the two cell populations CD133+CD45+CD34+ high and CD34+CD45+ high with a higher percentage. So, is the inclusion of Ac CD133 necessary in the pres-ent panel included in the ISHAGE methodflLast part, we showed a signif icant presence of interferon γ in CB in comparison to PAPB, the annexin showing the high number of apoptotic cells in CB. CONCLUSION: This study demonstrates that many different obstetric factors must be taken into account when processing and cryo-banking umbilical CB units for transplantation.

Suitability and limitations of mesenchymal stem cells to elucidate human bone illness

Functional impairment of mesenchymal stem cells(MSCs),osteoblast progenitor cells,has been proposed to be a pathological mechanism contributing to bone disorders,such as osteoporosis(the most common bone disease)and other rare inherited skeletal dysplasias.Pathological bone loss can be caused not only by an enhanced bone resorption activity but also by hampered osteogenic differentiation of MSCs.The majority of the current treatment options counteract bone loss,and therefore bone fragility by blocking bone resorption.These socalled antiresorptive treatments,in spite of being effective at reducing fracture risk,cannot be administered for extended periods due to security concerns.Therefore,there is a real need to develop osteoanabolic therapies to promote bone formation.Human MSCs emerge as a suitable tool to study the etiology of bone disorders at the cellular level as well as to be used for cell therapy purposes for bone diseases.This review will focus on the most relevant findings using human MSCs as an in vitro cell model to unravel pathological bone mechanisms and the application and outcomes of human MSCs in cell therapy clinical trials for bone disease.

Crosstalk between Autophagy and Apoptosis in Intervertebral Disc Degeneration

Objective: To describe the relationship between autophagy and apoptosis and the possible signaling pathways involved in degenerative lumbar intervertebral disc. Summary of Background Data: Autophagy and apoptosis are regulatory cellular mechanisms that determine many pathologies, including degenerative intervertebral disc disease. The interactions between these events in the damage or protection of intervertebral disc cells and in cellular homeostasis remain controversial. Methods: The sample size was twenty patients who underwent lumbar spine surgery for symptomatic disc herniation or spondylolisthesis. Intervertebral discs were classified by magnetic resonance as Pfirrmann grade IV and grade V. Six patients were operated on two levels, resulting in twenty-six intervertebral discs that were submitted to immunohistochemistry to verify the protein expression of autophagy and apoptosis markers. Results: The autophagic markers had greater protein expression in the human intervertebral disc (Pfirrmann Grades IV and V). Under these conditions, autophagy and apoptosis showed a negative correlation. Regarding apoptosis, caspase 8 presented the highest protein expression, which allows inferring the preference for the extrinsic pathway in cell death. Conclusions: Autophagy had the greatest protein expression negative profile compared to apoptosis. Caspase 8 had the highest protein expression in apoptosis.

Genotoxicity Clues to Predict Intervertebral Disc Degeneration: A Systematic Review

Objective: To characterize the association between DNA damage and Intervertebral disc degeneration (IDD). Summary of Background Data: IDD is the main disorder causing low back pain and is the most promising target for intervention. Many factors can contribute to the etiology, such as genetics, environment and lifestyle, but it is not yet fully understood. DNA damage can influence this process and needs to be studied, as well as the agents that can determine these damages. Methods: A systematic literature search of PubMed, Web of Science and Scopus was performed to identify studies related to DNA damage to the intervertebral disc. Results: After screening 61 records, 7 articles were included according to the selection criteria. All studies showed some relation between DNA damage and IDD. However, DNA damage was always considered a secondary issue to be investigated. Conclusions: Many factors can influence DNA damage induced by different genotoxic agents on the degenerative cascade of IVD. However, the correlation between IDD severity and DNA damage, as well as the factual role of DNA damage in disc degeneration could not be defined.

Towards on the Mediators in Collagen Replacement: Effect of the Active Molecules into a Polymeric Biomaterial

Modulating healing factors could avoid or minimize some possible pathological processes in collagen deposition. The present study was aimed to evaluated the role of active biomolecules such as PDGF-BB and PRP loaded or not into polymeric biomaterial to seek potential mediators in types I and III collagen deposition and epithelization. The healing phases were investigated by using an in vivo full-thickness wound rat model. At zero, 3rd, 7th and 14th days after the experimental model, the size of the wound areas was photographed. The nanofibrous materials were biocompatible and did not cause any local adverse reaction and/or inflammation. On day 14 the wounds had healed almost 100% with better signs of healing, however there was no obvious difference in the wound contraction rates. At the end of 14 days, samples from the center of the lesion were collected when histological features and immunopositivity for collagen I and III expressions were assessed. There was no significant difference in the epithelization among the groups. Wounds treated with PRP and with PA-6/SOMA plus PDGF-BB had significantly lower amounts of type III collagen. The amounts of type I collagen did not have a statistically different deposition among the experimental groups. The association of PDGF-BB with PA-6/SOMA emerges as an alternative for topical application to unfavorable anatomical sites, suggesting that these associations may have a positive modulation on the process of accelerated healing remodeling.

Synergistic effect of Mucuna pruriens and Withania somnifera in a paraquat induced Parkinsonian mouse model

Parkinson’s disease (PD) is a neurodegenerative disorder characterized by the development of rigidity, resting tremors and postural instability. Recently, the focus of PD’s treatment has shifted towards herbal medicines. Mucuna pruriens (Mp) and Withania somnifera (Ws) are traditional herbal medicines known to have neuro-protective effects due to the L-DOPA present in Mp seed powder and withanoloides present in Ws root extract. Here, the synergistic effect of Mp and Ws in Parkinsonian mice induced by chronic exposure to paraquat was evaluated. Co-treatment with Mp and Ws for 9 weeks, significantly decreased the elevated nitrite levels and lipid peroxidation found in Parkinsonian mice. In behavioural tests, Mp and Ws treated mice showed a significant decrease in the time taken to cross a narrow beam, an increase in the time of stay on drum in rotarod test and an improvement in the hanging time. Furthermore, it was found that the use of Mp and Ws considerably improved the tyrosine hydroxylase expression in the substantianigra region of the brain. The results suggest that Mp and Ws may provide a platform for future drug discoveries and novel treatment strategies for PD.

Cadaveric bone marrow mesenchymal stem cells:first experience treating a patient with large severe burns

Background:In January 2005,Rasulov et al.originally published"First experience in the use of bone marrow mesenchymal stem cells(MSCs)for the treatment of a patient with deep skin burns".Here,we present the first ever treated patient with cadaveric bone marrow mesenchymal stem cells(CMSCs)in the history of Medicine.Methods:A young man,who severely burned 60%of his total body surface with 30%of full-thickness burns while working with a grass trimmer that exploded,was involved in the study.MSCs were obtained from the bone marrow of a cadaver donor in a routine procurement procedure of CUCAIBA,the Province of Buenos Aires,Argentina,Ministry of Health,Transplantation Agency,cultured,expanded,and applied on the burned surfaces using a fibrin spray after early escharotomy.Results:So far,our preliminary experience and our early results have been very impressive showing an outstanding safety data as well as some impressive good results in the use of CMSCs.Conclusions:Based on al this,we think that improvements in the use of stem cells for burns might be possible in the near future and a lot of time as well as many lives could be saved by many other research teams all over the world.CMSCs will probably be a real scientific opportunity in Regenerative Medicine as well as in Transplantation.