Search results for: human umbilical cord
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8163

Search results for: human umbilical cord

8163 Umbilical Cord-Derived Cells in Corneal Epithelial Regeneration

Authors: Hasan Mahmud Reza

Abstract:

Extensive studies of the human umbilical cord, both basic and translational, over the last three decades have unveiled a plethora of information. The cord lining harbors at least two phenotypically different multipotent stem cells: mesenchymal stem cells (MSCs) and cord lining epithelial stem cells (CLECs). These cells exhibit a mixed genetic profiling of both embryonic and adult stem cells, hence display a broader stem features than cells from other sources. We have observed that umbilical cord-derived cells are immunologically privileged and non-tumorigenic by animal study. These cells are ethically acceptable, thus provides a significant advantage over other stem cells. The high proliferative capacity, viability, differentiation potential, and superior harvest of these cells have made them better candidates in comparison to contemporary adult stem cells. Following 30 replication cycles, these cells have been observed to retain their stemness, with their phenotype and karyotype intact. Transplantation of bioengineered CLEC sheets in limbal stem cell-deficient rabbit eyes resulted in regeneration of clear cornea with phenotypic expression of the normal cornea-specific epithelial cytokeratin markers. The striking features of low immunogenicity protecting self along with co-transplanted allografts from rejection largely define the transplantation potential of umbilical cord-derived stem cells.

Keywords: cord lining epithelial stem cells, mesenchymal stem cell, regenerative medicine, umbilical cord

Procedia PDF Downloads 113
8162 Effects of Umbilical Cord Clamping on Puppies Neonatal Vitality

Authors: Maria L. G. Lourenço, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

Abstract:

In veterinary medicine, the standard procedure during a caesarian section is clamping the umbilical cord immediately after birth. In human neonates, when the umbilical cord is kept intact after birth, blood continues to flow from the cord to the newborn, but this procedure may prove to be difficult in dogs due to the shorter umbilical cord and the number of newborns in the litter. However, a possible detachment of the placenta while keeping the umbilical cord intact may make the residual blood to flow to the neonate. This study compared the effects on neonatal vitality between clamping and no clamping the umbilical cord of dogs born through cesarean section, assessing them through Apgar and reflex scores. Fifty puppies delivered from 16 bitches were randomly allocated to receive clamping of the umbilical cord immediately (n=25) or to not receive the clamping until breathing (n=25). The neonates were assessed during the first five min of life and once again 10 min after the first assessment. The differences observed between the two moments were significant (p < 0.01) for both the Apgar and reflex scores. The differences observed between the groups (clamped vs. not clamped) were not significant for the Apgar score in the 1st moment (p=0.1), but the 2nd moment was significantly (p < 0.01) in the group not clamped, as well as significant (p < 0.05) for the reflex score in the 1st moment and 2nd moment (p < 0.05), revealing higher neonatal vitality in the not clamped group. The differences observed between the moments (1st vs. 2nd) of each group as significant (p < 0.01), revealing higher neonatal vitality in the 2nd moments. In the no clamping group, after removing the neonates together with the umbilical cord and the placenta, we observed that the umbilical cords were full of blood at the time of birth and later became whitish and collapsed, demonstrating the blood transfer. The results suggest that keeping the umbilical cord intact for at least three minutes after the onset breathing is not detrimental and may contribute to increase neonate vitality in puppies delivered by cesarean section.

Keywords: puppy vitality, newborn dog, cesarean section, Apgar score

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8161 In vitro Regeneration of Neural Cells Using Human Umbilical Cord Derived Mesenchymal Stem Cells

Authors: Urvi Panwar, Kanchan Mishra, Kanjaksha Ghosh, ShankerLal Kothari

Abstract:

Background: Day-by-day the increasing prevalence of neurodegenerative diseases have become a global issue to manage them by medical sciences. The adult neural stem cells are rare and require an invasive and painful procedure to obtain it from central nervous system. Mesenchymal stem cell (MSCs) therapies have shown remarkable application in treatment of various cell injuries and cell loss. MSCs can be derived from various sources like adult tissues, human bone marrow, umbilical cord blood and cord tissue. MSCs have similar proliferation and differentiation capability, but the human umbilical cord-derived mesenchymal stem cells (hUCMSCs) are proved to be more beneficial with respect to cell procurement, differentiation to other cells, preservation, and transplantation. Material and method: Human umbilical cord is easily obtainable and non-controversial comparative to bone marrow and other adult tissues. The umbilical cord can be collected after delivery of baby, and its tissue can be cultured using explant culture method. Cell culture medium such as DMEMF12+10% FBS and DMEMF12+Neural growth factors (bFGF, human noggin, B27) with antibiotics (Streptomycin/Gentamycin) were used to culture and differentiate mesenchymal stem cells into neural cells, respectively. The characterisations of MSCs were done with Flow Cytometer for surface markers CD90, CD73 and CD105 and colony forming unit assay. The differentiated various neural cells will be characterised by fluorescence markers for neurons, astrocytes, and oligodendrocytes; quantitative PCR for genes Nestin and NeuroD1 and Western blotting technique for gap43 protein. Result and discussion: The high quality and number of MSCs were isolated from human umbilical cord via explant culture method. The obtained MSCs were differentiated into neural cells like neurons, astrocytes and oligodendrocytes. The differentiated neural cells can be used to treat neural injuries and neural cell loss by delivering cells by non-invasive administration via cerebrospinal fluid (CSF) or blood. Moreover, the MSCs can also be directly delivered to different injured sites where they differentiate into neural cells. Therefore, human umbilical cord is demonstrated to be an inexpensive and easily available source for MSCs. Moreover, the hUCMSCs can be a potential source for neural cell therapies and neural cell regeneration for neural cell injuries and neural cell loss. This new way of research will be helpful to treat and manage neural cell damages and neurodegenerative diseases like Alzheimer and Parkinson. Still the study has a long way to go but it is a promising approach for many neural disorders for which at present no satisfactory management is available.

Keywords: bone marrow, cell therapy, explant culture method, flow cytometer, human umbilical cord, mesenchymal stem cells, neurodegenerative diseases, neuroprotective, regeneration

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8160 Maternal and Neonatal Outcome Analysis in Preterm Abdominal Delivery Underwent Umbilical Cord Milking Compared to Early Cord Clamping

Authors: Herlangga Pramaditya, Agus Sulistyono, Risa Etika, Budiono Budiono, Alvin Saputra

Abstract:

Preterm birth and anemia of prematurity are the most common cause of morbidity and mortality in neonates, and anemia of the preterm neonates has become a major issue. The timing of umbilical cord clamping after a baby is born determines the amount of blood transferred from the placenta to fetus, Delayed Cord Clamping (DCC) has proven to prevent anemia in the neonates but it is constrained concern regarding the delayed in neonatal resuscitation. Umbilical Cord Milking (UCM) could be an alternative method for clamping the umbilical cord due to the active blood transfer from the placenta to the fetus. The aim of this study was to analyze the difference between maternal and neonatal outcome in preterm abdominal delivery who underwent UCM compared to ECC. This was an experimental study with randomized post-test only control design. Analyzed maternal and neonatal outcomes, significant P values (P <0.05). Statistical comparison was carried out using Paired Samples t-test (α two tailed 0,05). The result was the mean of preoperative mother’s hemoglobin in UCM group compared to ECC (10,9 + 0,9 g/dL vs 10,4 + 0,9 g/dL) and postoperative (11,1 + 1,1 g/dL vs 10,5 + 0,7 g/dL), the delta was (0,2 + 0,7 vs 0,1 + 0,6.). It showed no significant difference (P=0,395 vs 0,627). The mean of 3rd phase labor duration in UCM group vs ECC was (20,5 + 3,5 second vs 21,1 + 3,3 second), showed insignificant difference (P=0,634). The amount of bleeding after delivery in UCM group compared to ECC has the median of 190 cc (100-280cc) vs 210 cc (150-330 cc) showed insignificant difference (P=0,083) so the incidence of post-partum bleeding was not found. The mean of the neonates hemoglobin, hematocrit and erythrocytes of UCM group compared to ECC was (19,3 + 0,7 vs 15,9 + 0,8 g/dl), (57,1 + 3,6 % vs 47,2 + 2,8 %), and (5,4 + 0,4 g/dl vs 4,5 + 0,3 g/dl) showed significant difference (P<0,0001). There was no baby in UCM group received blood transfusion and one baby in the control ECC group received blood transfusion was found. Umbilical Cord Milking has shown to increase the baby’s blood component such as hemoglobin, hematocrit, and erythrocytes 6 hours after birth as well as lowering the incidence of blood transfusions. Maternal and neonatal morbidity were not found. Umbilical Cord Milking was the act of clamping the umbilical cord that was more beneficial to the baby and no adverse or negative effects on the mother.

Keywords: umbilical cord milking, early cord clamping, maternal and neonatal outcome, preterm, abdominal delivery

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8159 Culture of Human Mesenchymal Stem Cells Culture in Xeno-Free Serum-Free Culture Conditions on Laminin-521

Authors: Halima Albalushi, Mohadese Boroojerdi, Murtadha Alkhabori

Abstract:

Introduction: Maintenance of stem cell properties during culture necessitates the recreation of the natural cell niche. Studies reported the promising outcome of mesenchymal stem cells (MSC) properties maintenance after using extracellular matrix such as CELLstart™, which is the recommended coating material for stem cells cultured in serum-free and xeno-free conditions. Laminin-521 is known as a crucial adhesion protein, which is found in natural stem cell niche, and plays an important role in facilitating the maintenance of self-renewal, pluripotency, standard morphology, and karyotype of human pluripotent stem cells (PSCs). The aim of this study is to investigate the effects of Laminin-521 on human umbilical cord-derived mesenchymal stem cells (UC-MSC) characteristics as a step toward clinical application. Methods: Human MSC were isolated from the umbilical cord via the explant method. Umbilical cord-derived-MSC were cultured in serum-free and xeno-free conditions in the presence of Laminin-521 for six passages. Cultured cells were evaluated by morphology and expansion index for each passage. Phenotypic characterization of UC-MSCs cultured on Laminin-521 was evaluated by assessment of cell surface markers. Results: Umbilical cord derived-MSCs formed small colonies and expanded as a homogeneous monolayer when cultured on Laminin-521. Umbilical cord derived-MSCs reached confluence after 4 days in culture. No statistically significant difference was detected in all passages when comparing the expansion index of UC-MSCs cultured on LN-521 and CELLstart™. Phenotypic characterization of UC-MSCs cultured on LN-521 using flow cytometry revealed positive expression of CD73, CD90, CD105 and negative expression of CD34, CD45, CD19, CD14 and HLA-DR.Conclusion: Laminin-521 is comparable to CELLstart™ in supporting UC-MSCs expansion and maintaining their characteristics during culture in xeno-free and serum-free culture conditions.

Keywords: mesenchymal stem cells, culture, laminin-521, xeno-free serum-free

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8158 A pilot Study of Umbilical Cord Mini-Clamp

Authors: Seng Sing Tan

Abstract:

Clamping of the umbilical cord after birth is widely practiced as a part of labor management. Further improvements were proposed to produce a smaller, lighter and more comfortable clamp while still maintaining current standards of clamping. A detachable holder was also developed to facilitate the clamping process. This pilot study on the efficacy of the mini-clamp was conducted to evaluate a tightness of the seal and a firm grip of the clamp on the umbilical cord. The study was carried out at National University Hospital, using 5 sets of placental cord. 18 samples of approximate 10 cm each were harvested. The test results showed that the mini-clamp was able to stop the flow through the cord after clamping without rupturing the cord. All slip tests passed with a load of 0.2 kg. In the pressure testing, 30kPa of saline was exerted into the umbilical veins. Although there was no physical sign of fluid leaking through the end secured by the mini-clamp, the results showed the pressure was not able to sustain the pressure set during the tests. 12 out of the 18 test samples have more than 7% of pressure drop in 30 seconds. During the pressure leak test, it was observed on several samples that when pressurized, small droplets of saline were growing on the outer surface of the cord lining membrane. It was thus hypothesized that the pressure drop was likely caused by the perfusion of the injected saline through the Wharton’s jelly and the cord lining membrane. The average pressure in the umbilical vein is roughly 2.67kPa (20 mmHg), less than 10% of 30kPa (~225mmHg), set for the pressure testing. As such, the pressure set could be over-specified, leading to undesirable outcomes. The development of the mini-clamp was an attempt to increase the comfort of newly born babies while maintaining the usability and efficacy of hospital grade umbilical cord clamp. The pressure leak in this study would be unfair to fully attribute it to the design and efficacy of the mini-clamp. Considering the unexpected leakage of saline through the umbilical membrane due to over-specified pressure exerted on the umbilical veins, improvements can definitely be made to the existing experimental setup to obtain a more accurate and conclusive outcome. If proven conclusive and effective, the mini-clamp with a detachable holder could be a smaller and potentially cheaper alternative to existing umbilical cord clamps. In addition, future clinical trials could be conducted to determine the user-friendliness of the mini-clamp and evaluate its practicality in the clinical setting by labor ward clinicians. A further potential improvement could be proposed on the sustainability factor of the mini-clamp. A biodegradable clamp would revolutionise the industry in this increasingly environmentally sustainability world.

Keywords: leak test, mini-clamp, slip test, umbilical cord

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8157 Correlation between Fetal Umbilical Cord pH and the Day, the Time and the Team Hand over Times: An Analysis of 6929 Deliveries of the Ulm University Hospital

Authors: Sabine Pau, Sophia Volz, Emanuel Bauer, Amelie De Gregorio, Frank Reister, Wolfgang Janni, Florian Ebner

Abstract:

Purpose: The umbilical cord pH is a well evaluated contributor for prediction of neonatal outcome. This study correlates nenonatal umbilical cord pH with the weekday of delivery, the time of birth as well as the staff hand over times (midwifes and doctors). Material and Methods: This retrospective study included all deliveries of a 20 year period (1994-2014) at our primary obstetric center. All deliveries with a newborn cord pH under 7,20 were included in this analysis (6929 of 48974 deliveries (14,4%)). Further subgroups were formed according to the pH (< 7,05; 7,05 – 7,09; 7,10 – 7,14; 7,15 – 7,19). The data were then separated in day- and night time (8am-8pm/8pm-8am) for a first analysis. Finally, handover times were defined at 6 am – 6.30 am, 2 pm -2.30 pm, 10 pm- 10.30 pm (midwives) and for the doctors 8-8.30 am, 4 – 4.30 pm (Monday- Thursday); 2 pm -2.30 pm (Friday) and 9 am – 9.30 am (weekend). Routinely a shift consists of at least three doctors as well as three midwives. Results: During the last 20 years, 6929 neonates were born with an umbilical cord ph < 7,20 ( < 7,05 : 7,1%; 7,05 – 7,09 : 10,9%; 7,10 – 7,14 : 30,2%; 7,15 – 7,19:51,8%). There was no significant difference between either night/day delivery (p = 0.408), delivery on different weekdays (p = 0.253), delivery between Monday to Thursday, Friday and the weekend (p = 0.496 ) or delivery during the handover times of the doctors as well as the midwives (p = 0.221). Even the standard deviation showed no differences between the groups. Conclusion: Despite an increased workload over the last 20 years, the standard of care remains high even during the handover times and night shifts. This applies for midwives and doctors. As the neonatal outcome depends on various factors, further studies are necessary to take more factors influencing the fetal outcome into consideration. In order to maintain this high standard of care, an adaption of work-load and changing conditions is necessary.

Keywords: delivery, fetal umbilical cord pH, day time, hand over times

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8156 Use of 3D Printed Bioscaffolds from Decellularized Umbilical Cord for Cartilage Regeneration

Authors: Tayyaba Bari, Muhammad Hamza Anjum, Samra Kanwal, Fakhera Ikram

Abstract:

Osteoarthritis, a degenerative condition, affects more than 213 million individuals globally. Since articular cartilage has no or limited vessels, therefore, after deteriorating, it is unable to rejuvenate. Traditional approaches for cartilage repair, like autologous chondrocyte implantation, microfracture and cartilage transplantation are often associated with postoperative complications and lead to further degradation. Decellularized human umbilical cord has gained interest as a viable treatment for cartilage repair. Decellularization removes all cellular contents as well as debris, leaving a biologically active 3D network known as extracellular matrix (ECM). This matrix is biodegradable, non-immunogenic and provides a microenvironment for homeostasis, growth and repair. UC derived bioink function as 3D scaffolding material, not only mediates cell-matrix interactions but also adherence, proliferation and propagation of cells for 3D organoids. This study comprises different physical, chemical and biological approaches to optimize the decellularization of human umbilical cord (UC) tissues followed by the solubilization of these tissues to bioink formation. The decellularization process consisted of two cycles of freeze thaw where the umbilical cord at -20˚C was thawed at room temperature followed by dissection in small sections from 0.5 to 1cm. Similarly decellularization with ionic and non-ionic detergents Sodium dodecyl sulfate (SDS) and Triton-X 100 revealed that both concentrations of SDS i.e 0.1% and 1% were effective in complete removal of cells from the small UC tissues. The results of decellularization was further confirmed by running them on 1% agarose gel. Histological analysis revealed the efficacy of decellularization, which involves paraffin embedded samples of 4μm processed for Hematoxylin-eosin-safran and 4,6-diamidino-2-phenylindole (DAPI). ECM preservation was confirmed by Alcian Blue, and Masson’s trichrome staining on consecutive sections and images were obtained. Sulfated GAG’s content were determined by 1,9-dimethyl-methylene blue (DMMB) assay, similarly collagen quantification was done by hydroxy proline assay. This 3D bioengineered scaffold will provide a typical atmosphere as in the extracellular matrix of the tissue, which would be seeded with the mesenchymal cells to generate the desired 3D ink for in vitro and in vivo cartilage regeneration applications.

Keywords: umbilical cord, 3d printing, bioink, tissue engineering, cartilage regeneration

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8155 Cord Blood Hematopoietic Stem Cell Expansion Ability of Mesenchymal Stem Cells Isolated From Different Sources

Authors: Ana M. Lara, Manuela Llano, Felipe Gaitán, Rosa H. Bustos, Ana Maria Perdomo-Arciniegas, Ximena Bonilla

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Umbilical cord blood is used as a source of progenitor and stem cells for the regeneration of the hematopoietic and immune system to treat patients with different hematological or non-hematological diseases. This stem cell source represents an advantage over the use of bone marrow or mobilized peripheral blood because it has a lower incidence rate of graft-versus-host disease, probably due to fewer immunological compatibility restrictions. However, its low cellular dose limits its use in pediatric patients. This work proposes the standardization of a cell expansion technique to compensate for the dose of infused cells through the ex-vivo manipulation of hematopoietic progenitor cells from umbilical cord blood before transplantation. The expansion model is carried out through co-cultures with mesenchymal stem cells (MSC) from bone marrow (BM) and less explored fetal tissues such as Wharton's jelly (WJ) and umbilical cord blood (UCB). Initially, a master cell bank of primary mesenchymal stem cells isolated from different sources was established and characterized following International Society of Cell Therapies (ISCT) indications. Additionally, we assessed the effect of a short 25 Gy cycle of gamma irradiation on cell cycle arrest of mesenchymal cells over the support capacity for the expansion of hematopoietic stem cells from umbilical cord blood was evaluated. The results show that co-cultures with MSC from WJ and UCB allow the cellular dose of HSPC to be maximized between 5 and 16 times having a similar support capacity as BM. In addition, was evaluated the hematopoietic stem progenitor cell's HSPC functionality through the evaluation of migration capacity, their differentiation capacity during culture time by flow cytometry to evaluate the expression of membrane markers associated with lineage-committed progenitors, their clonogenic potential, and the evaluation of secretome profile in the expansion process was evaluated. So far, the treatment with gamma irradiation maintains the hematopoietic support capacity of mesenchymal stem cells from the three sources studied compared to treatments without irradiation, favoring the use of fetal tissues that are generally waste to obtain mesenchymal cell lines for ex-vivo expansion systems. With the results obtained, a standardized protocol that will contribute to the development of ex-vivo expansion with MSC on a larger scale will be achieved, enabling its clinical use and expanding its application in adults.

Keywords: ex-vivo expansion, hematopoietic stem cells, hematopoietic stem cell transplantation, mesenchymal stem cells, umbilical cord blood

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8154 Comparative Study for Neonatal Outcome and Umbilical Cord Blood Gas Parameters in Balanced and Inhalant Anesthesia for Elective Cesarean Section in Dogs

Authors: Agnieszka Antończyk, MałGorzata Ochota, Wojciech Niżański, ZdzisłAw Kiełbowicz

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The goal of the cesarean section (CS) is the delivery of healthy, vigorous pups with the provision of surgical plane anesthesia, appropriate analgesia, and rapid recovery of the dam. In human medicine, spinal or epidural anesthesia is preferred for a cesarean section as associated with a lower risk of neonatal asphyxia and the need for resuscitation. Nevertheless, the specificity of veterinary patients makes the application of regional anesthesia as a sole technique impractical, thus to obtain patient compliance the general anesthesia is required. This study aimed to compare the influence of balanced (inhalant with epidural) and inhalant anesthesia on neonatal umbilical cord blood gas (UCBG) parameters and vitality (modified Apgar scoring). The bitches (31) undergoing elective CS were enrolled in this study. All females received a single dose of 0.2 mg/kg s.c. Meloxicam. Females were randomly assigned into two groups: Gr I (Isoflurane, n=16) and Gr IE (Isoflurane plus Epidural, n=15). Anesthesia was induced with propofol at 4-6 mg/kg to effect, and maintained with isoflurane in oxygen; in IE group epidural anesthesia was also done using lidocaine (3-4 mg/kg) into the lumbosacral space. CSs were performed using a standard mid-line approach. Directly after the puppy extraction, the umbilical cord was double clamped before the placenta detachment. The vessels were gently stretched between forceps to allow blood sampling. At least 100 mcl of mixed umbilical cord blood was collected into a heparinized syringe for further analysis. The modified Apgar scoring system (AS) was used to objectively score neonatal health and vitality immediately after birth (before first aid or neonatal care was instituted), at 5 and 20 min after birth. The neonates were scored as normal (AS 7-10), weak (AS 4-6), or critical (AS 0-3). During surgery, the IE group required a lower isoflurane concentration compared to the females in group I (MAC 1.05±0.2 and 1.4±0.13, respectively, p<0.01). All investigated UCBG parameters were not statistically different between groups. All pups had mild acidosis (pH 7.21±0.08 and 7.21±0.09 in Gr I and IE, respectively) with moderately elevated pCO2 (Gr I 57.18±11.48, Gr IE 58.74±15.07), HCO3- on the lower border (Gr I 22.58±3.24, Gr IE 22.83±3.6), lowered BE (Gr I -6.1±3.57, Gr IE -5.6±4.19) and mildly elevated level of lactates (Gr I 2.58±1.48, Gr IE2.53±1.03). The glucose levels were above the reference limits in both groups of puppies (74.50±25.32 in Gr I, 79.50±29.73 in Gr IE). The initial Apgar score results were similar in I and IE groups. However, the subsequent measurements of AS revealed significant differences between both groups. Puppies from the IE group received better AS scores at 5 and 20 min compared to the I group (6.86±2.23 and 8.06±2.06 vs 5.11±2.40 and 7.83±2.05, respectively). The obtained results demonstrated that administration of epidural anesthesia reduced the requirement for isoflurane in dams undergoing cesarean section and did not affect the neonatal umbilical blood gas results. Moreover, newborns from the epidural anesthesia group were scored significantly higher in AS at 5 and 20 min, indicating their better vitality and quicker improvement post-surgery.

Keywords: apgar scoring, balanced anesthesia, cesarean section, umbilical blood gas

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8153 Usage of Cord Blood Stem Cells of Asphyxia Infants for Treatment

Authors: Ahmad Shah Farhat

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Background: Prenatal asphyxia or birth asphyxia is the medical situation resulting from a newborn infant that lasts long enough during the birth process to cause physical harm, usually to the brain. Human umbilical cord blood (UCB) is a well-established source of hematopoietic stem/progenitor cells (HSPCs) for allogeneic stem cell transplantation. These can be used clinically to care for children with malignant diseases. Low O2 can cause in proliferation and differentiation of stem cells. Method: the cord blood of 11 infants with 3-5 Apgar scores or need to cardiac pulmonary Resuscitation as an asphyxia group and ten normal infants with more than 8 Apgar scores as the normal group was collected, and after isolating hematopoietic stem cells, the cells were cultured in enriched media for 14 days to compare the numbers of colonies by microscope. Results: There was a significant difference in the number of RBC precursor colonies (red colonies) in cultured media with 107 cord blood hematopoietic stem cells of infants who were exposed to hypoxemia in two wells of palate. There was not a significant difference in the number of white cell colonies in the two groups in the two wells of the plate. Conclusion: Hypoxia in the perinatal period can cause the increase of hematopoietic stem cells of cord blood, special red precursor stem cells in vitro, like an increase of red blood cells in the body when exposed to low oxygen conditions. Thus, it will be usable.

Keywords: asphyxia, neonre, stem cell, red cell

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8152 Umbilical Epidermal Inclusion Cysts, a Rare Cause of Umbilical Mass: A Case Report and Review of Literature

Authors: Christine Li, Amanda Robertson

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Epidermal inclusion cysts occur when epidermal cells are implanted in the dermis following trauma, or surgery. They are a rare cause of an umbilical mass, with very few cases previously reported following abdominal surgery. These lesions can present with a range of symptoms, including palpable mass, pain, redness, or discharge. This paper reports a case of an umbilical epidermal inclusion cyst in a 52-year-old female presenting with a six-week history of a painful, red umbilical lump on a background of two previous diagnostic laparoscopies. Abdominal computed tomography (CT) scans revealed non-specific soft tissue thickening in the umbilical region. This was successfully treated with complete excision of the lesion. Umbilical lumps are a common presentation but can represent a diagnostic challenge. The differential diagnosis should include an epidermal inclusion cyst, particularly in a patient who has had previous abdominal surgery, including laparoscopic surgery.

Keywords: epidermal inclusion cyst, laparoscopy, umbilical mass, umbilicus

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8151 Comparative Stem Cells Therapy for Regeneration of Liver Fibrosis

Authors: H. M. Imam, H. M. Rezk, A. F. Tohamy

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Background: Human umbilical cord blood (HUCB) is considered as a unique source for stem cells. HUCB contain different types of progenitor cells which could differentiate into hepatocytes. Aims: To investigate the potential of rat's liver damage repair using human umbilical cord mesenchymal stem cells (hUCMSCs). We investigated the feasibility for hUCMSCs in recovery from liver damage. Moreover, investigating fibrotic liver repair and using the CCl4-induced model for liver damage in the rat. Methods: Rats were injected with 0.5 ml/kg CCl4 to induce liver damage and progressive liver fibrosis. hUCMSCs were injected into the rats through the tail vein; Stem cells were transplanted at a dose of 1×106 cells/rat after 72 hours of CCl4 injection without receiving any immunosuppressant. After (6 and 8 weeks) of transplantation, blood samples were collected to assess liver functions (ALT, AST, GGT and ALB) and level of Procollagen III as a liver fibrosis marker. In addition, hepatic tissue regeneration was assessed histopathologically and immunohistochemically using antihuman monoclonal antibodies against CD34, CK19 and albumin. Results: Biochemical and histopathological analysis showed significantly increased recovery from liver damage in the transplanted group. In addition, HUCB stem cells transdifferentiated into functional hepatocytes in rats with hepatic injury which results in improving liver structure and function. Conclusion: Our findings suggest that transplantation of hUCMSCs may be a novel therapeutic approach for treating liver fibrosis. Therefore, hUCMSCs are a potential option for treatment of liver cirrhosis.

Keywords: carbon tetra chloride, liver fibrosis, mesenchymal stem cells, rat

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8150 Safety Study of Intravenously Administered Human Cord Blood Stem Cells in the Treatment of Symptoms Related to Chronic Inflammation

Authors: Brian M. Mehling, Louis Quartararo, Marine Manvelyan, Paul Wang, Dong-Cheng Wu

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Numerous investigations suggest that Mesenchymal Stem Cells (MSCs) in general represent a valuable tool for therapy of symptoms related to chronic inflammatory diseases. Blue Horizon Stem Cell Therapy Program is a leading provider of adult and children’s stem cell therapies. Uniquely we have safely and efficiently treated more than 600 patients with documenting each procedure. The purpose of our study is primarily to monitor the immune response in order to validate the safety of intravenous infusion of human umbilical cord blood derived MSCs (UC-MSCs), and secondly, to evaluate effects on biomarkers associated with chronic inflammation. Nine patients were treated for conditions associated with chronic inflammation and for the purpose of anti-aging. They have been given one intravenous infusion of UC-MSCs. Our study of blood test markers of 9 patients with chronic inflammation before and within three months after MSCs treatment demonstrates that there is no significant changes and MSCs treatment was safe for the patients. Analysis of different indicators of chronic inflammation and aging included in initial, 24-hours, two weeks and three months protocols showed that stem cell treatment was safe for the patients; there were no adverse reactions. Moreover data from follow up protocols demonstrates significant improvement in energy level, hair, nails growth and skin conditions. Intravenously administered UC-MSCs were safe and effective in the improvement of symptoms related to chronic inflammation. Further close monitoring and inclusion of more patients are necessary to fully characterize the advantages of UC-MSCs application in treatment of symptoms related to chronic inflammation.

Keywords: chronic inflammatory diseases, intravenous infusion, stem cell therapy, umbilical cord blood derived mesenchymal stem cells (UC-MSCs)

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8149 Comparison Study of 70% Ethanol Effect on Direct and Retrival Culture of Contaminated Umblical Cord Tissue for Expansion of Mesenchymal Stem Cells

Authors: Ganeshkumar, Ashika, Valavan, Ramesh, Thangam, Chirayu

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MSCs are found in much higher concentration in the Wharton’s jelly compared to the umbilical cord blood, which is a rich source of hematopoietic stem cells. Umbilical cord tissue is collected at the time of birth; it is processed and stored in liquid nitrogen for future therapeutical purpose. The source of contamination might be either from vaginal tract of mother or from hospital environment or from personal handling during cord tissue sample collection. If the sample were contaminated, decontamination procedure will be done with 70% ethanol (1 minute) in order to avoid sample rejection. Ethanol is effective against a wide range of bacteria, protozoa and fungi and has low toxicity to humans. Among the 1954 samples taken for the study, 24 samples were found to be contaminated with microorganism. The organisms isolated from the positive samples were found to be E. coli, Stenotrophomonas maltophilia, Pseudomonas aueroginosa, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, Enterobacter cloacae, and Proteus mirabilis. Among these organisms 70% ethanol successfully eliminated E. coli, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, and Proteus mirabilis. 70% ethanol was unsuccessful in eliminating Stenotrophomonas maltophilia, Pseudomonas aueroginosa, and Enterobacter cloacae. Stenotrophomonas maltophilia and Pseudomonas aueroginosa have the ability to form biofilm that make them resistant to alcohol. Biofilm act as protective layer for bacteria and which protects them from host defense and antibiotic wash. Finally it was found 70% ethanol wash saved 58.3% cord tissue samples from rejection and it is ineffective against 41% of the samples. The contamination rate can be reduced by maintaining proper aseptic techniques during sample collection and processing.

Keywords: umblical cord tissue, decontamination, 70% ethanol effectiveness, contamination

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8148 Treatment of Full-Thickness Rotator Cuff Tendon Tear Using Umbilical Cord Blood-Derived Mesenchymal Stem Cells and Polydeoxyribonucleotides in a Rabbit Model

Authors: Sang Chul Lee, Gi-Young Park, Dong Rak Kwon

Abstract:

Objective: The aim of this study was to investigate regenerative effects of ultrasound (US)-guided injection with human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) and/or polydeoxyribonucleotide (PDRN) injection in a chronic traumatic full-thickness rotator cuff tendon tear (FTRCTT) in a rabbit model. Material and Methods: Rabbits (n = 32) were allocated into 4 groups. After a 5-mm sized FTRCTT just proximal to the insertion site on the subscapularis tendon was created by excision, the wound was immediately covered by silicone tube to prevent natural healing. After 6 weeks, 4 injections (0.2 mL normal saline, G1; 0.2 mL PDRN, G2; 0.2 mL UCB-MSCs, G3; and 0.2 mL UCB-MSCs with 0.2ml PDRN, G4) were injected into FTRCTT under US guidance. We evaluated gross morphologic changes on all rabbits after sacrifice. Masson’s trichrome, anti-type 1 collagen antibody, bromodeoxyuridine, proliferating cell nuclear antigen, vascular endothelial growth factor and platelet endothelial cell adhesion molecule stain were performed to evaluate histological changes. Motion analysis was also performed. Results: The gross morphologic mean tendon tear size in G3 and 4 was significantly smaller than that of G1 and 2 (p < .05). However, there were no significant differences in tendon tear size between G3 and 4. In G4, newly regenerated collagen type 1 fibers, proliferating cells activity, angiogenesis, walking distance, fast walking time, and mean walking speed were greater than in the other three groups on histological examination and motion analysis. Conclusion: Co-injection of UCB-MSCs and PDRN was more effective than UCB-MSCs injection alone in histological and motion analysis in a rabbit model of chronic traumatic FTRCTT. However, there was no significant difference in gross morphologic change of tendon tear between UCB-MSCs with/without PDRN injection. The results of this study regarding the combination of UCB-MSCs and PDRN are worth additional investigations.

Keywords: mesenchymal stem cell, umbilical cord, polydeoxyribonucleotides, shoulder, rotator cuff, ultrasonography, injections

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8147 Genetic Diversity of Cord Blood of the National Center of Blood Transfusion, Mexico (NCBT)

Authors: J. Manuel Bello-López, Julieta Rojo-Medina

Abstract:

Introduction: The transplant of Umbilical Cord Blood Units (UCBU) are a therapeutic possibility for patients with oncohaematological disorders, especially in children. In Mexico, 48.5% of oncological diseases in children 1-4 years old are leukemias; whereas in patients 5-14 and 15-24 years old, lymphomas and leukemias represent the second and third cause of death in these groups respectively. Therefore it is necessary to have more registries of UCBU in order to ensure genetic diversity in the country; the above because the search for appropriate a UCBU is increasingly difficult for patients of mixed ethnicity. Objective: To estimate the genetic diversity (polymorphisms) of Human Leucocyte Antigen (HLA) Class I (A, B) and Class II (DRB1) in UCBU cryopreserved for transplant at Cord Blood Bank of the NCBT. Material and Methods: HLA typing of 533 UCBU for transplant was performed from 2003-2012 at the Histocompatibility Laboratory from the Research Department (evaluated by Los Angeles Ca. Immunogenetics Center) of the NCBT. Class I HLA-A, HLA-B and Class II HLA-DRB1 typing was performed using medium resolution Sequence-Specific Primer (SSP). In cases of an ambiguity detected by SSP; Sequence-Specific Oligonucleotide (SSO) method was carried out. A strict analysis of populations genetic parameters were done in 5 representative UCBU populations. Results: 46.5% of UCBU were collected from Mexico City, State of Mexico (30.95%), Puebla (8.06%), Morelos (6.37%) and Veracruz (3.37%). The remaining UCBU (4.75%) are represented by other states. The identified genotypes correspond to Amerindian origins (HLA-A*02, 31; HLA-B*39, 15, 48), Caucasian (HLA-A*02, 68, 01, 30, 31; HLA-B*35, 15, 40, 44, 07 y HLA-DRB1*04, 08, 07, 15, 03, 14), Oriental (HLA-A*02, 30, 01, 31; HLA-B* 35, 39, 15, 40, 44, 07,48 y HLA-DRB1*04, 07,15, 03) and African (HLA-A*30 y HLA-DRB1*03). The genetic distances obtained by Cavalli-Sforza analysis of the five states showed significant genetic differences by comparing genetic frequencies. The shortest genetic distance exists between Mexico City and the state of Puebla (0.0039) and the largest between Veracruz and Morelos (0.0084). In order to identify significant differences between this states, the ANOVA test was performed. This demonstrates that UCBU is significantly different according to their origin (P <0.05). This is shown by the divergence between arms at the Dendogram of Neighbor-Joining. Conclusions: The NCBT provides UCBU in patients with oncohaematological disorders in all the country. There is a group of patients for which not compatible UCBU can be find due to the mixed ethnic origin. For example, the population of northern Mexico is mostly Caucasian. Most of the NCBT donors are of various ethnic origins, predominantly Amerindians and Caucasians; although some ethnic minorities like Oriental, African and pure Indian ethnics are not represented. The NCBT is, therefore, establishing agreements with different states of Mexico to promote the altruistic donation of Umbilical Cord Blood in order to enrich the genetic diversity in its files.

Keywords: cord blood, genetic diversity, human leucocyte antigen, transplant

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8146 Leptin Levels in Cord Blood and Their Associations with the Birth of Small, Large and Appropriate for Gestational Age Infants in Southern Sri Lanka

Authors: R. P. Hewawasam, M. H. A. D. de Silva, M. A. G. Iresha

Abstract:

In recent years childhood obesity has increased to pan-epidemic proportions along with a concomitant increase in obesity-associated morbidity. Birth weight is an important determinant of later adult health, with neonates at both ends of the birth weight spectrum at risk of future health complications. Consequently, infants who are born large for gestational age (LGA) are more likely to be obese in childhood and adolescence and are at risk of cardiovascular and metabolic complications later in life. Adipose tissue plays a role in linking events in fetal growth to the subsequent development of adult diseases. In addition to its role as a storage depot for fat, adipose tissue produces and secrets a number of hormones of importance in modulating metabolism and energy homeostasis. Cord blood leptin level has been positively correlated with fetal adiposity at birth. It is established that Asians have lower skeletal muscle mass, low bone mineral content and excess body fat for a given body mass index indicating a genetic predisposition in the occurrence of obesity. To our knowledge, studies have never been conducted in Sri Lanka to determine the relationship between adipocytokine profile in cord blood and anthropometric parameters in newborns. Thus, the objective of this study is to establish the above relationship for the Sri Lankan population to implement awareness programs to minimize childhood obesity in the future. Umbilical cord blood was collected from 90 newborns (Male 40, Female 50; gestational age 35-42 weeks) after double clamping the umbilical cord before separation of the placenta and the concentration of leptin was measured by ELISA technique. Anthropometric parameters of the newborn such as birth weight, length, ponderal index, occipital frontal, chest, hip and calf circumferences were measured. Pearson’s correlation was used to assess the relationship between leptin and anthropometric parameters while the Mann-Whitney U test was used to assess the differences in cord blood leptin levels between small for gestational age (SGA), appropriate for gestational age (AGA) and LGA infants. There was a significant difference (P < 0.05) between the cord blood leptin concentrations of LGA infants (12.67 ng/mL ± 2.34) and AGA infants (7.10 ng/mL ± 0.90). However, a significant difference was not observed between leptin levels of SGA infants (8.86 ng/mL ± 0.70) and AGA infants. In both male and female neonates, umbilical leptin levels showed significant positive correlations (P < 0.05) with birth weight of the newborn, pre-pregnancy maternal weight and pre pregnancy BMI between the infants of large and appropriate for gestational ages. Increased concentrations of leptin levels in the cord blood of large for gestational age infants suggest that they may be involved in regulating fetal growth. Leptin concentration of Sri Lankan population was not significantly deviated from published data of Asian populations. Fetal leptin may be an important predictor of neonatal adiposity; however, interventional studies are required to assess its impact on the possible risk of childhood obesity.

Keywords: appropriate for gestational age, childhood obesity, leptin, anthropometry

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8145 Studying the Antiapoptotic Activity of Β Cells from Cord Blood Based Mesenchymal Stem Cells as an Approach to Treat Diabetes Mellitus

Authors: Parcha Sreenivasa Rao, P. Lakshmi

Abstract:

Diabetes Mellitus is metabolic disorder, characterized by high glucose levels in the blood due to one of the reason i.e., the death of β cells. The lack of β cells leads to the reduced insulin levels. The β cell death generally occurs due to apoptosis induced by the several cytokines. IL-1β, IFN- ϒ and TNF –α cytokines that are generally cause apoptosis to the β cell. The nutrient based apoptosis is generally seen with high glucose and free fatty acids. It is also noted that the β cell death triggered by Fas ligand and its receptor Fas at the surface of the activated CD8+ T- lymphocytes. Reports also reveal that the β cell apoptosis is under control of the transcription factors NF-kB and STAT- 1. The arresting or opposing of the β cell apoptosis can be overcome by the different growth factors like GLP-1, growth hormone, prolactin, VEGF, Dipeptidyl peptidase-4, Vildagliptin, suberoylanilidehydroxamic acid, trichistatin-A, XIAP, Bcl-2, FGF-21. Present investigation explains antiapoptotic property of the β cells derived from the mesenchymal stem cells of umbilical cord.

Keywords: stem cells, umblical cord, diabetes, apoptosis

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8144 Personalized Tissues and Organs Replacement – a Peek into the Future

Authors: Asaf Toker

Abstract:

Matricelf developed a technology that enables the production of autologous engineered tissue composed of matrix and cells derived from patients Omentum biopsy. The platform showed remarkable pre-clinical results for several medical conditions. The company recently licensed the technology that enabled scientist at Tel Aviv university that 3D printed a human heart from human cells and matrix for the first time in human history. The company plans to conduct its first human clinical trial for Acute Spinal Cord Injury (SCI) early in 2023.

Keywords: tissue engineering, regenerative medicine, spinal Cord Injury, autologous implants, iPSC

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8143 Morphometry of Cervical Spinal Cord in Rabbit Using Design-Based Stereology

Authors: Hamed Chavoshi Pour, Javad Sadeghinejad

Abstract:

The spinal cord is a long structure that starts at the end of the medulla oblongata and is located within the vertebral canal. Physiologically, the spinal cord connects the brain with the peripheral nervous system for sensory and motor activities. The cervical spinal cord is an area of particular interest in medicine and veterinary medicine due to the high prevalence of diseases in this region. This study describes the morphometric features of the cervical spinal cord in rabbits using design-unbiased stereology. The cervical spinal cords of five male rabbits were dissected, and slabs were taken according to systematic uniform random sampling. Each slab was embedded in paraffin and cut into a 6-µm thick section, and stained with cresyl violet 0.1% for stereological estimations. The total spinal cord volume, volume fraction of grey and white matter, and also dorsal and ventral horns were estimated using point counting and Cavalieri's estimator. The total cervical spinal cord volume was 0.98 ± 0.07 cm³. The relative volume of white matter and grey matter was 70.6 ± 1.7% and 29.31 ± 1.67%, respectively. The dorsal horn and ventral horn volume were 13.86 ± 1.36% and 14.9 ± 0.62% of the whole cervical spinal cord. This knowledge of rabbit spinal cord findings may serve as a foundation for a translational model in spinal cord experimental research and provide basic findings for the diagnosis and treatment of spinal cord disorders.

Keywords: stereology, spinal cord, rabbit, cervical

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8142 Effect of Maternal Factors and C-Peptide and Insulin Levels in Cord Blood on the Birth Weight of Newborns: A Preliminary Study from Southern Sri Lanka

Authors: M. H. A. D. de Silva, R. P. Hewawasam, M. A. G. Iresha

Abstract:

Macrosomia is common in infants born to not only women diagnosed with gestational diabetes mellitus but also non-diabetic obese women. Maternal Body Mass Index (BMI) correlates with the incidence of large for gestational age infants. Obesity has reached epidemic levels in modern societies. During the past two decades, obesity in children and adolescents has risen significantly in Asian populations including Sri Lanka. There is increasing evidence to believe that infants who are born large for gestational age are more likely to be obese in childhood and adolescence and are at risk of cardiovascular and metabolic complications later in life. It is also established that Asians have lower skeletal muscle mass, low bone mineral content and excess body fat for a given BMI indicating a genetic predisposition in the occurrence of obesity. The objective of this study is to determine the effect of maternal weight, weight gain during pregnancy, c-peptide and insulin concentrations in the cord blood on the birth of appropriate for and large for gestational age infants in a tertiary care center in Southern Sri Lanka. Umbilical cord blood was collected from 90 newborns (Male 40, Female 50; gestational age 35-42 weeks) after double clamping the umbilical cord before separation of the placenta and the concentration of insulin and C-peptide were measured by ELISA technique. Anthropometric parameters of the newborn such as birth weight, length, ponderal index, occipital frontal, chest, hip and calf circumferences were measured, and characteristics of the mother were collected. The relationship between insulin, C-peptide and anthropometrics were assessed by Spearman correlation. The multiple logistic regression analysis examined influences of maternal weight, weight gain during pregnancy, C-peptide and insulin concentrations in cord blood as covariates on the birth of large for gestational age infants. A significant difference (P<0.001) was observed between the insulin levels of infants born large for gestational age (18.73 ± 0.52 µlU/ml) and appropriate for gestational age (13.08 ± 0.56 µlU/ml). Consistently, A significant decrease in concentration (41.68%, P<0.001) was observed between C-peptide levels of infants born large for gestational age and appropriate for gestational age. Cord blood insulin and C-peptide levels had a significant correlation with birth weight (r=0.35, P<0.05) of the newborn at delivery. Maternal weight and BMI which are indicators of maternal nutrition were proven to be directly correlated with birth weight and length. To our knowledge, this relationship was investigated for the first time in a Sri Lankan setting and was also evident in our results. This study confirmed the fact that insulin and C-peptide play a major role in regulating fetal growth. According to the results obtained in this study, we can suggest that the increased BMI of the mother has a direct influence on increased maternal insulin secretion, which may subsequently affect cord insulin and C-peptide levels and also birth weight of the infant.

Keywords: C-peptide, insulin, large for gestational age, maternal weight

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8141 Research on Key Technologies on Initial Installation of Ultra-Deep-Water Dynamic Umbilical

Authors: Weiwei Xie, Yichao Li

Abstract:

The initial installation of the umbilical can affect the subsequent installation process and final installation. Meanwhile, the design of both ends of the ultra-deep water dynamic umbilical (UDWDU), as well as the design of the surface unit and the subsea production system connected by UDWDU,], varies in different oil and gas fields. To optimize the installation process of UDWDU, on the basis of the summary and analysis of the surface-end and the subsea-end design of UDWDU and the mainstream construction resources, the method of initial installation from the surface unit side or the subsea production system side of UDWDU is studied, and each initiation installation method is pointed out if some difficulties that may be encountered.

Keywords: dynamic umbilical, ultra-deep-water, initial installation, installation process

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8140 Efficacy of Umbilical Cord Lining Stem Cells For Wound Healing in Diabetic Murine Model

Authors: Fui Ping Lim, Wen Choong Chua, Toan Thang Phan

Abstract:

Aim: This study investigates the roles of Cord Lining Stem Cells (CLSCs) as potential therapeutic agents for diabetic wounds. Method: 20 genetically diabetic db/db mice were randomly assigned to two arms; (i) control group received placebo treatment (sham media or cells delivery material), and (ii) active comparator received CLSCs. Two full-thickness wounds, each sized 10mm X 10mm were created, one on each side of the midline on the back of the mice. Digital pictures were taken on day 1, 3, 7, 10, 14, 17, 21, 24, 28. Wound areas were analyzed with ImageJ TM software and calculated as percentage of the original wound. Time to closure was defined as the day the wound bed was completely epithelized and filled with new tissues. Results: The CLSCs-treated wounds, showed a significant increase in the percentage of wound closure and achieved 100% closure of the wound sooner than the control group by an average of 3.7 days. The mice treated with CLSCs have a shorter wound closure time (mean closure day: 19.8 days) as compared to the control group (mean closure day: 23.5 days). Conclusion: Our preliminary findings inferred that CLSCs treated wound achieved higher percentage of wound closure within a shorter duration of time.

Keywords: cord lining stem cell, diabetic wound, stem cell, wound

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8139 Application of Mesenchymal Stem Cells in Diabetic Therapy

Authors: K. J. Keerthi, Vasundhara Kamineni, A. Ravi Shanker, T. Rammurthy, A. Vijaya Lakshmi, Q. Hasan

Abstract:

Pancreatic β-cells are the predominant insulin-producing cell types within the Islets of Langerhans and insulin is the primary hormone which regulates carbohydrate and fat metabolism. Apoptosis of β-cells or insufficient insulin production leads to Diabetes Mellitus (DM). Current therapy for diabetes includes either medical management or insulin replacement and regular monitoring. Replacement of β- cells is an attractive treatment option for both Type-1 and Type-2 DM in view of the recent paper which indicates that β-cells apoptosis is the common underlying cause for both the Types of DM. With the development of Edmonton protocol, pancreatic β-cells allo-transplantation became possible, but this is still not considered as standard of care due to subsequent requirement of lifelong immunosuppression and the scarcity of suitable healthy organs to retrieve pancreatic β-cell. Fetal pancreatic cells from abortuses were developed as a possible therapeutic option for Diabetes, however, this posed several ethical issues. Hence, in the present study Mesenchymal stem cells (MSCs) were differentiated into insulin producing cells which were isolated from Human Umbilical cord (HUC) tissue. MSCs have already made their mark in the growing field of regenerative medicine, and their therapeutic worth has already been validated for a number of conditions. HUC samples were collected with prior informed consent as approved by the Institutional ethical committee. HUC (n=26) were processed using a combination of both mechanical and enzymatic (collagenase-II, 100 U/ml, Gibco ) methods to obtain MSCs which were cultured in-vitro in L-DMEM (Low glucose Dulbecco's Modified Eagle's Medium, Sigma, 4.5 mM glucose/L), 10% FBS in 5% CO2 incubator at 37°C. After reaching 80-90% confluency, MSCs were characterized with Flowcytometry and Immunocytochemistry for specific cell surface antigens. Cells expressed CD90+, CD73+, CD105+, CD34-, CD45-, HLA-DR-/Low and Vimentin+. These cells were differentiated to β-cells by using H-DMEM (High glucose Dulbecco's Modified Eagle's Medium,25 mM glucose/L, Gibco), β-Mercaptoethanol (0.1mM, Hi-Media), basic Fibroblast growth factor (10 µg /L,Gibco), and Nicotinamide (10 mmol/L, Hi-Media). Pancreatic β-cells were confirmed by positive Dithizone staining and were found to be functionally active as they released 8 IU/ml insulin on glucose stimulation. Isolating MSCs from usually discarded, abundantly available HUC tissue, expanding and differentiating to β-cells may be the most feasible cell therapy option for the millions of people suffering from DM globally.

Keywords: diabetes mellitus, human umbilical cord, mesenchymal stem cells, differentiation

Procedia PDF Downloads 228
8138 Non-linear Analysis of Spontaneous EEG After Spinal Cord Injury: An Experimental Study

Authors: Jiangbo Pu, Hanhui Xu, Yazhou Wang, Hongyan Cui, Yong Hu

Abstract:

Spinal cord injury (SCI) brings great negative influence to the patients and society. Neurological loss in human after SCI is a major challenge in clinical. Instead, neural regeneration could have been seen in animals after SCI, and such regeneration could be retarded by blocking neural plasticity pathways, showing the importance of neural plasticity in functional recovery. Here we used sample entropy as an indicator of nonlinear dynamical in the brain to quantify plasticity changes in spontaneous EEG recordings of rats before and after SCI. The results showed that the entropy values were increased after the injury during the recovery in one week. The increasing tendency of sample entropy values is consistent with that of behavioral evaluation scores. It is indicated the potential application of sample entropy analysis for the evaluation of neural plasticity in spinal cord injury rat model.

Keywords: spinal cord injury (SCI), sample entropy, nonlinear, complex system, firing pattern, EEG, spontaneous activity, Basso Beattie Bresnahan (BBB) score

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8137 Evaluation of Traumatic Spine by Magnetic Resonance Imaging

Authors: Sarita Magu, Deepak Singh

Abstract:

Study Design: This prospective study was conducted at the department of Radio Diagnosis, at Pt B.D. Sharma PGIMS, Rohtak in 57 patients of spine injury on radiographs or radiographically normal patients with neurological deficits presenting within 72 hours of injury. Aims: Evaluation of the role of Magnetic Resonance Imaging (MRI) in Spinal Trauma Patients and to compare MRI findings with clinical profile and neurological status of the patient and to correlate the MRI findings with neurological recovery of the patient and predict the outcome. Material and Methods: Neurological status of patients was assessed at the time of admission and discharge in all the patients and at long term interval of six months to one year in 27 patients as per American spine injury association classification (ASIA). On MRI cord injury was categorized into cord hemorrhage, cord contusion, cord edema only, and normal cord. Quantitative assessment of injury on MRI was done using mean canal compromise (MCC), mean spinal cord compression (MSCC) and lesion length. Neurological status at admission and neurological recovery at discharge and long term follow up was compared with various qualitative cord findings and quantitative parameters on MRI. Results: Cord edema and normal cord was associated with favorable neurological outcome. Cord contusion show lesser neurological recovery as compared to cord edema. Cord hemorrhage was associated with worst neurological status at admission and poor neurological recovery. Mean MCC, MSCC, and lesion length values were higher in patients presenting with ASIA A grade injury and showed decreasing trends towards ASIA E grade injury. Patients showing neurological recovery over the period of hospital stay and long term follow up had lower mean MCC, MSCC, and lesion length as compared to patients showing no neurological recovery. The data was statistically significant with p value <.05. Conclusion: Cord hemorrhage and higher MCC, MSCC and lesion length has poor prognostic value in spine injury patients.

Keywords: spine injury, cord hemorrhage, cord contusion, MCC, MSCC, lesion length, ASIA grading

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8136 Concentrations of Leptin, C-Peptide and Insulin in Cord Blood as Fetal Origins of Insulin Resistance and Their Effect on the Birth Weight of the Newborn

Authors: R. P. Hewawasam, M. H. A. D. de Silva, M. A. G. Iresha

Abstract:

Obesity is associated with an increased risk of developing insulin resistance. Insulin resistance often progresses to type-2 diabetes mellitus and is linked to a wide variety of other pathophysiological features including hypertension, hyperlipidemia, atherosclerosis (metabolic syndrome) and polycystic ovarian syndrome. Macrosomia is common in infants born to not only women with gestational diabetes mellitus but also non-diabetic obese women. During the past two decades, obesity in children and adolescents has risen significantly in Asian populations including Sri Lanka. There is increasing evidence to believe that infants who are born large for gestational age (LGA) are more likely to be obese in childhood. It is also established from previous studies that Asian populations have higher percentage body fat at a lower body mass index compared to Caucasians. High leptin levels in cord blood have been reported to correlate with fetal adiposity at birth. Previous studies have also shown that cord blood C-peptide and insulin levels are significantly and positively correlated with birth weight. Therefore, the objective of this preliminary study was to determine the relationship between parameters of fetal insulin resistance such as leptin, C-peptide and insulin and the birth weight of the newborn in a study population in Southern Sri Lanka. Umbilical cord blood was collected from 90 newborns and the concentration of insulin, leptin, and C-peptide were measured by ELISA technique. Birth weight, length, occipital frontal, chest, hip and calf circumferences of newborns were measured and characteristics of the mother such as age, height, weight before pregnancy and weight gain were collected. The relationship between insulin, leptin, C-peptide, and anthropometrics were assessed by Pearson’s correlation while the Mann-Whitney U test was used to assess the differences in cord blood leptin, C-peptide, and insulin levels between groups. A significant difference (p < 0.001) was observed between the insulin levels of infants born LGA (18.73 ± 0.64 µlU/ml) and AGA (13.08 ± 0.43 µlU/ml). Consistently, A significant increase in concentration (p < 0.001) was observed in C-peptide levels of infants born LGA (9.32 ± 0.77 ng/ml) compared to AGA (5.44 ± 0.19 ng/ml). Cord blood leptin concentration of LGA infants (12.67 ng/mL ± 1.62) was significantly higher (p < 0.001) compared to the AGA infants (7.10 ng/mL ± 0.97). Significant positive correlations (p < 0.05) were observed among cord leptin levels and the birth weight, pre-pregnancy maternal weight and BMI between the infants of AGA and LGA. Consistently, a significant positive correlation (p < 0.05) was observed between the birth weight and the C peptide concentration. Significantly high concentrations of leptin, C-peptide and insulin levels in the cord blood of LGA infants suggest that they may be involved in regulating fetal growth. Although previous studies suggest comparatively high levels of body fat in the Asian population, values obtained in this study are not significantly different from values previously reported from Caucasian populations. According to this preliminary study, maternal pre-pregnancy BMI and weight may contribute as significant indicators of cord blood parameters of insulin resistance and possibly the birth weight of the newborn.

Keywords: large for gestational age, leptin, C-peptide, insulin

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8135 Factors Contributing to the Risk and Vulnerability to HIV Infection among Individuals with Spinal Cord Injuries (SCI) in South Africa

Authors: J. J. Lloyd, J. S. Phillips

Abstract:

Background: HIV/AIDS has made a huge impact on human development and sexual reproductive habits in this century in the world and especially in sub-Saharan Africa. It has only recently been acknowledged that HIV/AIDS has an equal if not greater effect on or threat to people with disabilities. Survivors of traumatic spinal cord injury (SCI) with resultant disability are incorrectly believed to be sexually inactive, unlikely to use drugs or alcohol and at less risk of violence or rape than their non-disabled peers. This group can thus be described as economically, educationally and socially disadvantaged, which in itself, suggest that they are a high-risk group for HIV infection. Objectives: Thus, the overall objective of this study was to assess the factors that exacerbate the risk and vulnerability of individuals with spinal cord injuries to HIV infection in order to develop a more effective HIV intervention. Methodology: This paper reports on the cross-sectional data gathered from individuals with a traumatic spinal cord injury in 4 conveniently selected provinces in South Africa. Data was collected by means of self-administered questionnaires. The questionnaire consisted of various sections requesting for information on Demographics; HIV-Knowledge (HIV- KQ-18); Sexual behaviours; sexual communication, and negotiation skills and Self-efficacy to refuse sex. Results: The majority of the study sample was males (72.7%) with a mean age of 34.6 years. The majority reported lifetime sexual intercourse (92.4%) but only 31.8% reported condom use with last sexual intercourse. Low level of HIV knowledge, and being male were the strongest predictor of risky sexual behaviours in this sample. Conclusion: Significant numbers of individuals with spinal cord injuries are thus engaging in risky sexual behaviours pointing to a need to strengthen comprehensive sexual health education to increase access to HIV testing, promote safe sex and condom use among this group.

Keywords: Human Immunodeficiency Virus (HIV), individuals with spinal cord injuries, risky sexual behaviours, HIV risk factors, sub-saharan Africa

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8134 A Forearm-Wrist Rehabilitation Module for Stroke and Spinal Cord Injuries

Authors: Vahid Mehrabi, Iman Sharifi, H. A. Talebi

Abstract:

The automation of rehabilitation procedure by the implementation of robotic devices can overcome the limitation in conventional physiotherapy methods by increasing training sessions and duration of process. In this paper, the design of a simple rehabilitation robot for forearm-wrist therapy in stroke and spinal cord injuries is presented. Wrist’s biological joint motion is modeled by a gimbal-like mechanism which resembles the human arm anatomy. Presented device is an exoskeleton robot with rotation axes corresponding to human skeleton anatomy. The mechanical structure, actuator and sensor selection, system kinematics and comparison between our device range of motion and required active daily life values is illustrated.

Keywords: rehabilitation, robotic devices, physiotherapy, forearm-wrist

Procedia PDF Downloads 238