Plasma-Based Scaffold Containing Bone-Marrow Mononuclear Cells Promotes Wound Healing in a Mouse Model of Pressure Injury.

Pressure injuries, or pressure ulcers, are a common problem that may lead to infections and major complications, besides being a social and economic burden due to the costs of treatment and hospitalization. While surgery is sometimes necessary, this also has complications such as recurrence or wound dehiscence. Among the newer methods of pressure injury treatment, advanced therapies are an interesting option. This study examines the healing properties of bone marrow mononuclear cells (BM-MNCs) embedded in a plasma-based scaffold in a mouse model. Pressure ulcers were created on the backs of mice (2 per mouse) using magnets and assigned to a group of ulcers that were left untreated (Control, n = 15), treated with plasma scaffold (Plasma, n = 15), or treated with plasma scaffold containing BM-MNC (Plasma + BM-MNC, n = 15). Each group was examined at three time points (3, 7, and 14 days) after the onset of treatment. At each time point, animals were subjected to biometric assessment, bioluminescence imaging, and tomography. Once treatment had finished, skin biopsies were processed for histological and wound healing reverse transcription polymerase chain reaction (RT-PCR) array studies. While wound closure percentages were higher in the Plasma and Plasma + BM-MNC groups, differences were not significant, and thus descriptive data are provided. In all individuals, the presence of donor cells was revealed by immunohistochemistry on posttreatment onset Days 3, 7, and 14. In the Plasma + BM-MNC group, less inflammation was observed by positron emission tomography-computed tomography (PET/CT) imaging of the mice at 7 days, and a complete morphometabolic response was produced at 14 days, in accordance with histological results. A much more pronounced inflammatory process was observed in controls than in the other two groups, and this persisted until Day 14 after treatment onset. RT-PCR array gene expression patterns were also found to vary significantly, with the greatest difference noted between both treatments at 14 days when 11 genes were differentially expressed.

Stem cell and tissue engineering approaches in pressure ulcer treatment.

CONTEXT: Pressure ulcers or injuries, arise from ischemic damage to soft tissues induced by unrelieved pressure over a bony prominence. They are usually difficult to treat with standard medical therapy and often they recur. In the search for better treatment options, promising alternative forms of treatment are today emerging. Within the field of regenerative medicine, ongoing research on advanced therapies seeks to develop medicinal products based on gene therapy, somatic cell therapy, tissue-engineering and combinations of these. OBJECTIVE: The main objective is to perform an overview of experimental and clinical developments in somatic cell therapy and tissue engineering targeting the treatment of pressure injuries. METHODS: Searching terms as "PRESSURE ULCER", "STEM CELL THERAPY", "TISSUE ENGINEERING" or "WOUND HEALING" were used in combination or alone, including publications refered to basic and clinical research and focusing on articles showing results obtained in a clinical context. A total of 80 references are cited, including 23 references published in the 3 last years. RESULTS: The results suggest that this form of treatment could be an interesting option in patients with difficult-to-treat ulcers as spinal cord injury patients. CONCLUSION: This field of regenerative medicine is very broad and further research is warranted.

Treatment of pressure ulcers in patients with spinal cord injury: Conventional surgery vs. cellular therapy.

CONTEXT: Relapse and recurrence rates of pressure injuries (PIs) are very high in spinal cord injured patients. That is the reason why alternative therapies, such the stem cells derived from bone marrow, have been developed. OBJECTIVE: To compare this new technique of infiltration-infusion of mononuclear cells from bone marrow with conventional surgery. DESIGN: A retrospective study was carried out in patients with spinal cord injuries who had PIs, category III/IV, in the pelvic area, during a 14-year follow-up period. SETTING: One group was treated with conventional surgery and, in the other group, mononuclear cells were infused. PARTICIPANTS: One hundred and forty-nine patients were registered, 63 (42.3%) in the conventional surgery group and 86 (57.7%) in the mononuclear cell group. RESULTS: A comparative study between these 2 groups was carried out. There were no significant differences in ulcer healing in the first 6 months, but 6 months and one-year post-treatment, they were found. At 6 months, no patient in the conventional surgery group showed dehiscence or fistulization of the wound and, one year after surgery, only 3.17% recurred in the conventional group. In addition, there was a statistically significant relationship between days of hospitalization and the type of bacterial contamination and the intervention group. CONCLUSION: Bone marrow mononuclear cell infusion-infiltration is an alternative treatment for PIs and fistula during the first 6 months, instead of conventional surgery. However, in the medium-long term, conventional surgery is more effective.

The role of bone marrow mononuclear cell-conditioned medium in the proliferation and migration of human dermal fibroblasts.

BACKGROUND: Several recent studies have demonstrated the great potential of bone marrow cells in regenerative medicine, not only for their ability to differentiate to match a damaged cell type, but also because they synthesize and release various growth factors and cytokines.We examined the effect of bone marrow cell-conditioned medium in the healing process, especially in terms of fibroblast proliferation and migration. METHODS: These in vitro studies consisted of co-culture (without direct contact) of dermal fibroblasts with mononuclear bone marrow cells and the use of conditioned medium obtained from these cultures in a scratch wound model. RESULTS: Mononuclear cells were found to increase the proliferation of fibroblasts, and the conditioned medium showed a stimulatory effect on the migration of fibroblasts. CONCLUSION: When considered together with the observed increase in growth factor levels in conditioned medium, it appears that these cells act through a paracrine mechanism.

Opciones terapéuticas del embarazo ectópico sobre cicatriz de cesárea / Therapeutic options of ectopic pregnancy over cesarean scar

El embarazo ectópico sobre cicatriz de cesárea previa es una complicación obstétrica cuya incidencia está aumentando en los ultimos años debido al incremento de las tasas de parto por cesárea. Es fundamental un diagnóstico precoz para realizar un tratamiento adecuado y evitar las graves complicaciones que puede presentar. El principal problema que nos encontramos ante el abordaje de esta entidad es la ausencia de unas pautas claras para su tratamiento. Presentamos el caso de una paciente diagnosticada de embarazo ectópico sobre cicatriz de cesárea previa tratada con éxito con metotrexato intramuscular en dosis única, con una rápida resolución analítica y ecográfica

A caesarean scar pregnancy is an obstetric complication whose incidence is increased in recent years due to an increment of caesarean rates. It is critical to early diagnosis to make appropriate treatment and prevent serious complications that can present. The main problem that we are facing the approach of this entity is the absence of clear guidelines for treatment. We report the case of a patient diagnosed at 9+5 weeks of caesarean scar pregnancy that was treated in our center successfully with methotrexate intramuscularly in a single dose, with rapid analytical and ultrasonographic resolution

Platelet-Derived Mitochondria Display Embryonic Stem Cell Markers and Improve Pancreatic Islet ß-cell Function in Humans.

Diabetes is a major global health issue and the number of individuals with type 1 diabetes (T1D) and type 2 diabetes (T2D) increases annually across multiple populations. Research to develop a cure must overcome multiple immune dysfunctions and the shortage of pancreatic islet ß cells, but these challenges have proven intractable despite intensive research effort more than the past decades. Stem Cell Educator (SCE) therapy-which uses only autologous blood immune cells that are externally exposed to cord blood stem cells adhering to the SCE device, has previously been proven safe and effective in Chinese and Spanish subjects for the improvement of T1D, T2D, and other autoimmune diseases. Here, 4-year follow-up studies demonstrated the long-term safety and clinical efficacy of SCE therapy for the treatment of T1D and T2D. Mechanistic studies found that the nature of platelets was modulated in diabetic subjects after receiving SCE therapy. Platelets and their released mitochondria display immune tolerance-associated markers that can modulate the proliferation and function of immune cells. Notably, platelets also expressed embryonic stem cell- and pancreatic islet ß-cell-associated markers that are encoded by mitochondrial DNA. Using freshly-isolated human pancreatic islets, ex vivo studies established that platelet-releasing mitochondria can migrate to pancreatic islets and be taken up by islet ß cells, leading to the proliferation and enhancement of islet ß-cell functions. These findings reveal new mechanisms underlying SCE therapy and open up new avenues to improve the treatment of diabetes in clinics. Stem Cells Translational Medicine 2017;6:1684-1697.

Study of the interaction of GB virus C/Hepatitis G virus fusion peptides belonging to the E2 protein with phospholipid Langmuir monolayers.

In order to determine the ability of 1,2-dipalmitoyl phosphatidylcholine (DPPC) and 1,2-dioleoyl phosphatidylglycerol (DOPG) to host peptide sequences belonging to the E2 protein of GBV virus C/Hepatitis G virus, the behaviour of Langmuir monolayers formed by these phospholipids and E2 (12-26), E2 (354-363) and E2 (chimeric) peptide sequences was analysed from data of surface pressure (π) versus area per molecule (A) isotherms, compression modulus (Cs-1), excess Gibbs energy of mixing (ΔGexc) and total Gibbs energy of mixing (ΔGmix). Three different behaviours were observed. Mixed films of E2 (12-26) with DPPC or DOPC showed negative values for the excess thermodynamic functions, and thus attractive interactions between mixed films components are greater than in ideal films. Mixtures of E2 (354-363) with DPPC or DOPG, exhibited positive values of excess functions, evidencing weaker interactions in the mixed films in relation to those of pure components. Finally, positive and negative excess functions were observed in E2 (chimeric)/DPPC or DOPG mixed films, depending on their composition. In short, the interaction between the phospholipids used in this work as models of cell membranes and E2 peptides varies with the type of phospholipid and the nature of the peptide (size, bulky, hydrophobicity and electric charge).

Repair of long-bone pseudoarthrosis with autologous bone marrow mononuclear cells combined with allogenic bone graft.

BACKGROUND AIMS: Long-bone pseudoarthrosis is a major orthopedic concern because of numerous factors such as difficulty of the treatment, high recurrence, high costs and the devastating effects on the patients' quality of life, which sometimes ends in amputation. Although the "gold standard" for the treatment of this pathology is autologous bone grafting, which has high osteogenic, osteoconductive and osteoinductive properties, this treatment presents some restrictions such as the limited amount of bone that can be taken from the patient and donor site morbidity. Bone marrow mononuclear cells (BM-MNCs) comprise progenitor and stem cells with pro-angiogenic and pro-osteogenic properties. Allogenic cancellous bone graft is a natural and biodegradable osteoconductive and osteoinductive scaffold. Combination of these two components could mimic the advantages of autologous bone grafting while avoiding its main limitations. METHODS: Long-bone pseudoarthrosis was treated in seven patients with autologous BM-MNCs from iliac crest combined with frozen allogenic cancellous bone graft obtained from the tissue bank. RESULTS: All patients showed complete bone consolidation 5.3 ± 0.9 months (range, 2-9 months) after cell transplantation. Moreover, limb pain disappeared in all of them. The mean follow-up was 35.8 ± 4.6 months after transplantation (range, 24-51 months) without pseudoarthrosis recurrence or pain reappearing. CONCLUSIONS: Combination of autologous BM-MNCs and allogenic bone graft could constitute an easy, safe, inexpensive and efficacious attempt to treat long-bone pseudoarthrosis and non-union by reproducing the beneficial properties of autologous bone grafting while restricting its disadvantages.

Secuestro broncopulmonar fetal con resolución espontánea intraútero / Fetal bronchopulmonary sequestration with spontaneous intrauterine resolution

Presentamos un caso de secuestro broncopulmonar (SBP) sospechado en la ecografía morfológica de la semana 20 de gestación, debido al hallazgo de una imagen triangular ecogénica homogénea en el pulmón izquierdo del feto, que se confirma 2 semanas más tarde al detectar el vaso sistémico que irriga la lesión. Debido a la ausencia de factores de mal pronóstico, se decide mantener una actitud expectante, observándose una disminución progresiva de la lesión hasta su completa desaparición ecográfica en la semana 37 (AU)

We present a case of bronchopulmonary sequestration suspected in the morphological ultrasound at 20 weeks of gestation based on the finding of a homogeneous hyperechoic triangular image in the left lung of the fetus, which was confirmed 2 weeks later with the observation of a blood vessel from the aorta feeding the mass. Due to the absence of poor prognostic factors, an expectant attitude was adopted, with a progressive decrease of the mass until its complete disappearance on ultrasound at 37 weeks (AU)

Advances in stem cell therapy.

Since the beginning of stem cell biology, considerable effort has been focused in the translation of scientific insights into new therapies. Cell-based assays represent a new strategy for organ and tissue repair in several pathologies. Moreover, alternative treatment strategies are urgently needed due to donor organ shortage that costs many lives every year and results in lifelong immunosuppression. At the moment, only the use of hematopoietic stem cells is considered as the standard for the treatment of malignant and genetic bone marrow disorders, being all other stem cell applications highly experimental. The present chapter tries to summarize some ongoing approaches of stem cell regenerative medicine and also introduces recent findings from published studies and trials conducted in various tissues such as skeletal muscle, liver and lung.

Tumor de Krukenberg secundario a carcinoma de colon durante el embarazo / Krukenberg tumor secondary to carcinoma of the colon during pregnancy

El tumor de Krukenberg se define como aquel tumor metastásico en ovario uni o bilateral que contiene cantidades importantes de células en anillo de sello y cuyo origen es principalmente digestivo. La incidencia de tumores metastásicos en ovario es muy pequeña (1-5%); asimismo la coincidencia de una masa ovárica de características malignas metastásicas con una gestación es ínfima. Presentamos el caso de una mujer de 19 años que tras el diagnóstico de un adenocarcinoma mucinoso de colon, a los 18 meses presenta un tumor de Krukenberg en la semana 29 de gestación. En un primer momento se le realizó cesárea junto a cirugía resectiva del tumor. En un segundo tiempo se le practicó cirugía citorreductora asociada a quimioterapia intraperitoneal intraoperatoria hipertérmica, encontrándose actualmente en remisión clínica y en seguimiento por el servicio de oncología (AU)

Krukenberg tumor is defined as a metastatic uni- or bilateral ovarian tumor that contains significant amounts of signet ring cells and whose origin is mainly gastrointestinal. The incidence of metastatic tumors of the ovary is very small (1-5%), and that of the concurrence of an ovarian mass with metastatic malignant features and pregnancy is negligible. We report the case of a 19-year-old woman who, 18 months after diagnosis of a mucinous adenocarcinoma of the colon, was diagnosed with a Krukenberg tumor in the 29th week of pregnancy. Initially, cesarean section together with tumoral resection was performed. In a second stage, cytoreductive surgery was performed with hyperthermic intraoperative intraperitoneal chemotherapy. The patient is currently in clinical remission and is monitored by the oncology service (AU)

Metotrexato sistémico o intrasacular en el tratamiento del embarazo cervical / Systemic or intrasacular methotrexate in the treatment of cervical pregnancy

Se presenta el caso de una gestante a la que se le detectó un embarazo ectópico cervical en la sexta semana de gestación. La rapidez del diagnóstico y el buen estado clínico de la paciente permitieron la instauración de tratamiento conservador, siendo preciso el uso de dos dosis de metotrexato sistémico para que disminuyeran las cifras de la beta gonadotropina coriónica humana, y posteriormente el empleo de metotrexato intrasacular, debido a la persistencia de la imagen ecográfica. El tratamiento médico con metotrexato sistémico y/o intrasacular es una medida altamente eficaz para conseguir una escasa morbilidad, reservando otras técnicas más agresivas para aquellos casos que no respondan a este tratamiento o si aparecen complicaciones (AU)

We present a case of cervical ectopic pregnancy diagnosed in the sixth week of gestation. The speed of diagnosis and good clinical status of the patient allowed conservative treatment to be applied. Two doses of systemic methotrexate were required to decrease human chorionic gonadotrophin (BetaHCG) values followed by the use of intrasaccular methotrexate due to the persistence of the ultrasound image. Medical treatment with systemic and/or intrasaccular methotrexate is a highly effective measure to achieve low morbidity. This option allows more aggressive techniques to be reserved for patients unresponsive to this treatment or those with complications (AU)

Brain death effects on catecholamine levels and subsequent cardiac damage assessed in organ donors.

BACKGROUND: Brain death (BD) causes hemodynamic and neuroendocrine alterations including a catecholamine surge, which in turn causes histologic lesions in cardiac muscle such as contraction bands, focal mononuclear cell infiltrates and cardiomyocyte necrosis. These changes are likely to compromise heart function and could therefore also affect the graft response after heart transplantation. This study was designed to examine the catecholamine surge, the catecholamine release pattern and the histologic lesions traditionally described as characteristic of BD in hearts procured from BD donors. METHODS: After BD diagnosis, specimens were taken from the left ventricle (n = 50) for histologic examination. Arterial blood samples were collected from 40 of the donors at different time-points (1 hour before BD; on BD diagnosis; and 1, 2, 3 and 4 hours after BD) to determine catecholamine levels by high-performance liquid chromatography (HPLC). RESULTS: The three hormones examined showed above-normal levels (epinephrine 2.36-fold, norepinephrine 8.56-fold, dopamine 54.76-fold). Release patterns included epinephrine and dopamine peaks at the time of BD and a norepinephrine peak 1 hour later. Fifty percent of the BD donors showed contraction bands and 62% displayed cardiomyocyte necrosis, which was associated with focal mononuclear cell infiltrates in 18% of cases. In 40% of donors, colocalized apoptotic and necrotic damage was observed. CONCLUSIONS: Differing extents of BD-associated cardiac lesions were observed in the donors, and >50% also showed apoptotic damage. The expected catecholamine peak at the time of BD was only detected for epinephrine and dopamine. Hormone increases were below those described in the literature, except for dopamine.

Interaction of GB virus C/hepatitis G virus synthetic peptides with lipid langmuir monolayers and large unilamellar vesicles.

In this paper, we aimed to continue the previous study undertaken with one segment of E1 protein belonging to the GB virus C/hepatitis G virus (GBV-C/HGV), specifically between the 53-66 amino acids and their palmitoyl derivative peptide. The sequence selection has been made on the basis of different prediction algorithms of hydrophobicity and antigenicity. Their interactions between two different in vitro membrane models, lipid Langmuir monolayers and vesicles of different lipidic composition, have been evaluated. For this purpose, different lipids, varying the charge and the unsaturations of the hydrocarbon chain 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (sodium salt) (DPPG) and 1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (sodium salt) (DOPG), have been selected. Miscibility and peptides/lipids interactions have been analyzed on the basis of surface pressure (pi)-mean molecular area (A) isotherms, which have been recorded for pure and mixed monolayers of different composition spread at the air/water interface. Furthermore, E1(53-66) sequence and PalmE1(53-66) have been labeled with a fluorescent group, succinimidyl 6-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoate (NBD succinimide), in order to study their behavior in the presence of vesicles. The obtained results are consistent with the existence of electrostatic (attractive) intermolecular interactions between the two positive net charges of the peptides and the polar heads of negative-charged lipids. However, both the lipidic membrane fluidity and the palmitic chain linked to the native peptide play an important role in the balance between the electrostatic forces established at the interface and the hydrophobic ones established inside the membrane. The fluorescence assays have demonstrated that electrostatic forces clearly predominate over the hydrophobic interactions only when the native sequence is retained at the polar interface of DPPG and DOPG vesicles. However, the palmitic tail linked to the peptide helped its penetration in the hydrophobic environment of the membrane, and this process was favored by decreasing the membrane fluidity.

Behaviour of a peptide sequence from the GB virus C/hepatitis G virus E2 protein in Langmuir monolayers: its interaction with phospholipid membrane models.

The GB virus C/hepatitis G virus (GBV C/HGV) is a Flaviviridae member that despite its non pathogenicity, has become of great interest given that it could inhibit the replication of the human immunodeficiency virus (HIV). Therefore, a better knowledge of the virus peptides involved in the cellular membrane fusion mechanism has become our aim. The selected peptide, named E2(347-363), corresponds to the GBV-C/HGV E2 protein and has been synthesized in order to study its interaction with in vitro membrane models. Two phospholipids, varying the charge and the unsaturations of the hydrocarbon chain have been chosen: 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (sodium salt) (DOPG). For our porpoise, we have used the Langmuir monolayer technique and Brewster angle microscopy (BAM) to gain deeper insight into the peptide/lipid interactions. The results obtained allow us to argue in favour of considering E2(347-363) a success candidate for developing further experiments in order to determine its potential role in the GBV C/HGV virus/cell membrane fusion process.

A molecular approach to apoptosis in the human heart during brain death.

BACKGROUND: Brain death induces changes in tissues and organs destined for transplant at the cell, molecular, and endocrine level including cell death through apoptosis. This study was designed to examine apoptotic damage in cardiac tissue obtained from brain dead donors. METHODS: Fifty tissue specimens from the left ventricles of individual donors were processed to evaluate changes in the expression levels of five genes involved in apoptosis (BAX, BCL2, CASPASE 3, CYTOCHROME C, and FAS) using the real time-polymerase chain reaction technique. Expression levels were quantified by the relative standard method and results normalized to the levels recorded for the endogenous control peptidylprolyl isomerase A. The HIF1alpha gene was also determined to check for the possibility of hypoxic damage. Control ventricular tissue specimens were obtained from patients undergoing mitral valve replacement. RESULTS: Using a mixed linear model it was determined that the sample type (donor vs. control patient) significantly affected (P<0.0001) expression levels of the genes examined reflected by their Ct values. Three of the genes (BAX, CASPASE 3, and FAS) showed significantly higher (Student's t test, P<0.05) expression levels (4.89-, 7.85-, and 12.14-fold endogenous control values, respectively) in donors compared with control patients (2.31-, 2.64-, and 3.57-fold endogenous control values, respectively) indicating the activation of apoptosis during brain death. CONCLUSION: Our findings suggest the possibility of using antiapoptosis agents to prevent cardiac injury and improve posttransplant behavior.