Maternal Th17/Treg Cytokines and Small Extracellular Vesicles in Plasma as Potential Biomarkers for Preeclampsia.

Preeclampsia is one of the most serious pregnancy complications. It may be caused by immunological changes in the early placental microenvironment. The contents of small EVs may serve as biomarkers of pregnancy complications. Evidence suggests that the balance between T helper 17 (Th17) and regulatory T (Treg) cells are critical for preventing preeclampsia. The study recruited 39 pregnant women with preeclampsia and 127 healthy pregnant women. We assessed the levels of both Th17 and Treg cytokines (IL-10, IL-17, IL-21, IL-22, and TGF-ß) in their plasma and small EVs. We found significant differences in the levels of all cytokines in the plasma between the two groups during the second trimester. We also observed significant differences between the two groups in the levels of EV-encapsulated cytokines IL-21, IL-22, and TGF-ß, as well as in total small EVs, during the second trimester. The ROC analysis showed that the classification efficiency (AUC) of TGF-ß in small EVs was 0.81. TGF-ß had the best discriminant ability of all the single EV biomarkers tested, the cross-validation of the accuracy was 0.89. Th17 and Treg cytokines in plasma and small EVs may contribute to maternal immune activation and clarify the potential mechanisms of small EVs and cytokines in preeclampsia.

Hyaluronan Induces a Mitochondrial Functional Switch in Fast-Proliferating Human Mesenchymal Stem.

BACKGROUND AND OBJECTIVES: Hyaluronan preserves the proliferation and differentiation potential of mesenchymal stem cells. Supplementation of low-concentration hyaluronan (SHA) in stem cells culture medium increases its proliferative rate, whereas coated-surface hyaluronan (CHA) maintains cells in a slow-proliferating mode. We have previously demonstrated that in CHA, the metabolic proliferative state of stem cells was influenced by upregulating mitochondrial biogenesis and function. However, the effect of SHA on stem cells' energetic status remains unknown. In this study, we demonstrate the effect that low-concentration SHA at 0.001 mg/ml (SHA0.001) and high-concentration SHA at 5 mg/ml (SHA5) exert on stem cells' mitochondrial function compared with CHA and noncoated tissue culture surface (control). METHODS AND RESULTS: Fast-proliferating human placenta-derived mesenchymal stem cells (PDMSCs) cultured on SHA0.001 exhibited reduced mitochondrial mass, lower mitochondrial DNA copy number, and lower oxygen consumption rate compared with slow-proliferating PDMSCs cultured on CHA at 5.0 (CHA5) or 30 µg/cm2 (CHA30). The reduced mitochondrial biogenesis observed in SHA0.001 was accompanied by a 2-fold increased ATP content and lactate production, suggesting that hyaluronan-induced fast-proliferating PDMSCs may rely less on mitochondrial function as an energy source and induce a mitochondrial functional switch to glycolysis. CONCLUSIONS: PDMSCs cultured on both CHA and SHA exhibited a reduction in reactive oxygen species levels. The results from this study clarify our understandings on the effect of hyaluronan on stem cells and provide important insights into the effect of distinct supplementation methods used during cell therapies.

Hyaluronan keeps mesenchymal stem cells quiescent and maintains the differentiation potential over time.

Hyaluronan (HA), an abundant polysaccharide found in human bodies, plays a role in the mesenchymal stem cells (MSCs) maintenance. We had previously found that HA prolonged the lifespan, and prevented the cellular aging of murine adipose-derived stromal cells. Recently, we had also summarized the potential pathways associated with HA regulation in human MSCs. In this study, we used the human placenta-derived MSCs (PDMSC) to investigate the effectiveness of HA in maintaining the PDMSC. We found that coating the culture surface coated with 30 µg cm-2 of HA (C) led to cluster growth of PDMSC, and maintained a higher number of PDMSC in quiescence compared to those grown on the normal tissue culture surface (T). PDMSC were treated for either 4 (short-term) or 19 (long-term) consecutive passages. PDMSC which were treated with HA for 19 consecutive passages had reduced cell enlargement, preserved MSCs biomarker expressions and osteogenic potential when compared to those grown only on T. The PDMSC transferred to T condition after long-term HA treatment showed preserved replicative capability compared to those on only T. The telomerase activity of the HA-treated PDMSC was also higher than that of untreated PDMSC. These data suggested a connection between HA and MSC maintenance. We suggest that HA might be regulating the distribution of cytoskeletal proteins on cell spreading in the event of quiescence to preserve MSC stemness. Maintenance of MSCs stemness delayed cellular aging, leading to the anti-aging phenotype of PDMSC.

Hyaluronan Upregulates Mitochondrial Biogenesis and Reduces Adenoside Triphosphate Production for Efficient Mitochondrial Function in Slow-Proliferating Human Mesenchymal Stem Cells.

Hyaluronan-coated surfaces preserve the proliferation and differentiation potential of mesenchymal stem cells by prolonging their G1-phase transit, which maintains cells in a slow-proliferative mode. Mitochondria are known to play a crucial role in stem cell self-renewal and differentiation. In this study, for the first time, the metabolic mechanism underlying the hyaluronan-regulated slow-proliferative maintenance of stem cells was investigated by evaluating mitochondrial functions. Human placenta-derived mesenchymal stem cells (PDMSCs) cultured on hyaluronan-coated surfaces at 0.5, 3.0, 5.0, and 30 µg/cm2 were found to have an average 58% higher mitochondrial mass and an increase in mitochondrial DNA copy number compared to noncoated tissue culture surfaces (control), as well as a threefold increase in the gene expression of the mitochondrial biogenesis-related gene PGC-1α. Increase in mitochondrial biogenesis led to a hyaluronan dose-dependent increase in mitochondrial membrane potential, ATP content, and oxygen consumption rate, with reactive oxygen species levels shown to be at least three times lower compared to the control. Although hyaluronan seemed to favor mitochondrial function, cell entry into a hyaluronan-regulated slow-proliferative mode led to a fivefold reduction in ATP production and coupling efficiency levels. Together, these results suggest that hyaluronan-coated surfaces influence the metabolic proliferative state of stem cells by upregulating mitochondrial biogenesis and function with controlled ATP production. This more efficiently meets the energy requirements of slow-proliferating PDMSCs. A clear understanding of the metabolic mechanism induced by hyaluronan in stem cells will allow future applications that may overcome the current limitations faced in stem cell culture. Stem Cells 2016;34:2512-2524.

Differential Proteomic Analysis of Human Placenta-Derived Mesenchymal Stem Cells Cultured on Normal Tissue Culture Surface and Hyaluronan-Coated Surface.

Our previous results showed that hyaluronan (HA) preserved human placenta-derived mesenchymal stem cells (PDMSC) in a slow cell cycling mode similar to quiescence, the pristine state of stem cells in vivo, and HA was found to prevent murine adipose-derived mesenchymal stem cells from senescence. Here, stable isotope labeling by amino acid in cell culture (SILAC) proteomic profiling was used to evaluate the effects of HA on aging phenomenon in stem cells, comparing (1) old and young passage PDMSC cultured on normal tissue culture surface (TCS); (2) old passage on HA-coated surface (CHA) compared to TCS; (3) old and young passage on CHA. The results indicated that senescence-associated protein transgelin (TAGLN) was upregulated in old TCS. Protein CYR61, reportedly senescence-related, was downregulated in old CHA compared to old TCS. The SIRT1-interacting Nicotinamide phosphoribosyltransferase (NAMPT) increased by 2.23-fold in old CHA compared to old TCS, and is 0.48-fold lower in old TCS compared to young TCS. Results also indicated that components of endoplasmic reticulum associated degradation (ERAD) pathway were upregulated in old CHA compared to old TCS cells, potentially for overcoming stress to maintain cell function and suppress senescence. Our data points to pathways that may be targeted by HA to maintain stem cells youth.

Effects of music listening on stress, anxiety, and sleep quality for sleep-disturbed pregnant women.

Prenatal sleep disturbance has been associated with undesirable birthing outcomes. To determine the effectiveness of listening to music at home in improving sleep quality, 121 Taiwanese pregnant women with poor sleep quality (Pittsburgh Sleep Quality Index [PSQI] score > 5) were systematically assigned, with a random start to music listening (n = 61) or control (n = 60) group. Participants in the music listening group self-regulated listening to music in addition to receiving general prenatal care similar to that in the control group for 2 weeks. The PSQI and State-Anxiety Inventory were used to assess outcomes. ANCOVA analyses were used with the pretest scores as covariates and showed significant improvement in sleep quality, stress, and anxiety in the music listening group compared with the control group. The most frequently used music genre by participants in the experimental group was lullabies, followed by classical music and crystal baby music. This study supported the theory that 2-week music listening interventions may reduce stress, anxiety, and yield better sleep quality for sleep-disturbed pregnant women. The analysis of participants' journals also implied that the expectant mothers' choices of musical genres may correlate more with perceived prenatal benefits or the desire to interact with their unborn child.

The effects of music listening on psychosocial stress and maternal-fetal attachment during pregnancy.

OBJECTIVE: While music listening has been studied as an intervention to help reduce anxiety in pregnant women, few studies have explored the effect of music listening on pregnancy-specific stress relief. This study examines the effects of music listening on psychosocial stress and maternal-fetal attachment during pregnancy. DESIGN: A randomized controlled trial was implemented. A valid sample of 296 pregnant women in their second or third trimester was randomly distributed into an experimental group (n=145) and a control group (n=151). INTERVENTIONS: The experimental group received routine prenatal care and music listening. The control group received routine prenatal care only. MAIN OUTCOME MEASURES: Data were collected using a demographic form, Pregnancy Stress Rating Scale (PSRS), Perceived Stress Scale (PSS), and Maternal-Fetal Attachment Scale (MFAS). RESULTS: The post-test results identified a significantly lower level of psychosocial stress in the experimental group than in the control group, particularly in terms of the stresses related to baby care and changing family relationships and to maternal role identification. However, no statistically significant differences in terms of perceived stress and maternal-fetal attachment were found between the post-test results of the two groups. CONCLUSION: This study provides evidence in support of using of music in interventions designed to relieve psychosocial stress in prenatal women. IRB approval number: ER98223.

Prenatal sonographic diagnosis of single umbilical artery: Emphasis on the absent side and its relation to associated anomalies.

OBJECTIVE: To determine the absent side of a single umbilical artery (SUA) and to evaluate whether associated anomalies are related to the side of the missing artery in a Taiwanese population. MATERIALS AND METHODS: We retrospectively studied SUA fetuses from our computer database of fetal ultrasound in a tertiary medical center in Southern Taiwan. All cases were diagnosed as SUA prenatally using conventional scanners of two- and three-dimensional (2D and 3D, respectively) ultrasound, as well as color, power, and high-definition Doppler. The absent side of UA and associated anomalies were analyzed. RESULTS: From September 2006 to November 2011, 31 fetuses with SUA were diagnosed prenatally by ultrasound and all were enrolled for this series. The incidence was estimated to be 1:556 (0.18% = 31/17,086). The mean maternal age was 29.2 years (range, 15-36 years) and the mean fetal age was 30.0 weeks of gestation (range 18-36 weeks). Notably, the left-absent UA was detected in 16/31 (52%) fetuses, compared with the right-absent UA in 15/31 (48%) cases. In addition, congenital anomalies were noted prenatally in 2/16 (13%) fetuses with left-absent UA and in 3/15 (20%) fetuses with right-absent UA. CONCLUSION: In SUA fetuses, the absence of UA appears to occur equally at each side. Moreover, this study showed no significant difference between either side of missing UA and associated anomalies after statistical examination.

An association study of interleukin-4 gene and preeclampsia in Taiwan.

OBJECTIVE: To test whether the interleukin-4 (IL-4) gene polymorphisms could be used as markers of susceptibility for preeclampsia in Taiwan. MATERIALS AND METHODS: This study included 78 women with preeclampsia and 125 normal controls. A polymerase chain reaction was used to analyze the variable number of tandem repeats polymorphism for the IL-4 gene intron 3. Restriction fragment length polymorphism analysis using endonuclease BsmFI was performed for the IL-4 gene at the promoter -590 position. The association between the genotype and disease was examined using Chi-square tests. RESULTS: We found no significant differences in the genotype distributions and allele frequencies for the IL-4 gene at both promoter and intron 3 regions between the preeclampsia and control groups. CONCLUSION: Even though this is the first study investigating the association between the promoter region and intron 3 polymorphisms of IL-4 and preeclampsia worldwide, our data do not support a role of the IL-4 gene in the pathogenesis of preeclampsia in Taiwanese women.

On the decellularization of fresh or frozen human umbilical arteries: implications for small-diameter tissue engineered vascular grafts.

Most tissues, including those to be decellularized for tissue engineering applications, are frozen for long term preservation. Such conventional cryopreservation has been shown to alter the structure and mechanical properties of tissues. Little is known, however, how freezing affects decellularization of tissues. The purpose of this study was two-fold: to examine the effects of freezing on decellularization of human umbilical arteries (HUAs), which represent a potential scaffolding material for small-diameter tissue-engineered vascular grafts, and to examine how decellularization affects the mechanical properties of frozen HUAs. Among many decellularization methods, hypotonic sodium dodecyl sulfate solution was selected as the decellularizing agent and tested on fresh HUAs to optimize decellularization conditions. The efficiency of decellularization was evaluated by DNA assay and histology every 12 up to 48 h. The optimized decellularization protocol was then performed on frozen HUAs. The stiffness, burst pressure, and suture retention strength of fresh HUAs and frozen HUAs before and after decellularization were also examined. It appeared that freezing decreased the efficiency of decellularization, which may be attributed to the condensed extracellular matrix caused by freezing. While the stiffness of fresh HUAs did not change significantly after decellularization, decellularization reduced the compliance of frozen HUAs. Interestingly, the stiffness of decellularized frozen HUAs was similar to that of decellularized fresh HUAs. Although little difference in stiffness was observed, we suggest avoiding freezing if more efficient and complete decellularization is desired.

Prenatal diagnosis of fetal gastroschisis using three-dimensional ultrasound: comparison between the 20th and 21st centuries.

OBJECTIVE: In order to compare the trends and improvements of prenatal diagnosis of gastroschisis, we herein retrospectively reviewed our cases of fetal gastroschisis detected by three-dimensional ultrasound (3D US) between the two centuries. MATERIALS AND METHODS: We reviewed our computer database of prenatal diagnosis on gastroschisis in National Cheng Kung University Hospital from October 1994 to November 2011. All the fetuses were initially scanned by two-dimensional (2D) US to locate the region of interest (ROI). Then, the 3D probe was used to scan all the ROI systematically and mechanically, and all the images were stored on laser discs for further 3D visualization and reconstruction. To compare the characteristics at prenatal diagnosis of gastroschisis between the 20th and 21st centuries in our hospital, the Chi-square test and Student t test were used. The p values less than 0.05 and 0.1 were considered statistically significant. RESULTS: In total, 26 fetuses with gastroschisis were depicted by 3D US in utero (10 cases were diagnosed in the 20th century and 16 cases in the 21st century). The ranges of gestational age at prenatal diagnosis of gastroschisis by 3D US in the 20th century were between 14 and 34 weeks (mean: 21.6 weeks) and between 14 and 33 weeks (mean: 21.9 weeks) in the 21st century. Moreover, seven cases (70%) were diagnosed before the third trimester in the 20th century, whereas 13 cases (81%) were diagnosed before the third trimester in the 21st century. CONCLUSION: Although without statistical significance, higher prenatal diagnosis rate before the third trimester in the 21st century was noted. The improvement of 3D US has remarkable advantages in adding novel visual depiction of a 3D lesion of a 3D fetus in 3D US after reconstruction and thus assists substantially in prenatal diagnosis, genetic consultation, and perinatal management of gastroschisis.

Development and validation of the postpartum sleep quality scale.

BACKGROUND: Postpartum sleep disorders are a significant problem for postnatal women. Although factors predisposing postnatal women to sleep disorders have been identified, few sleep quality assessment instruments adequately address these factors. PURPOSE: This study aimed to develop a Postpartum Sleep Quality Scale (PSQS) and test its psychometric properties. METHODS: Sixteen PSQS items were generated from clinical practice, a literature review, and expert recommendations. Two hundred two postpartum women were recruited from a medical center and an obstetric clinic in southern Taiwan in 2010-2011 to assess the internal consistency, test-retest reliability, construct validity, and convergent validity of the developed PSQS. RESULTS: Item analysis removed two of the initial PSQS items. The resultant 14-item PSQS showed good internal consistency (α = .81) and acceptable 5-day test-retest reliability (r = .81). Construct validity was confirmed using exploratory factor analysis, which extracted and defined "Infant night care-related daytime dysfunction" and "Physical symptoms-related sleep inefficiency" as the two main categories of postpartum sleep quality. These two factors explained 44.49% of total variance, with factor loadings between .43 and .76. Significant correlation (r = .67) with the Pittsburgh Sleep Quality Index showed convergent validity. CONCLUSION: The 14-item PSQS is a reliable, valid, and useful scale for measuring postpartum sleep quality and examining intervention protocols in Taiwanese postpartum women with sleep disturbance.

Prenatal diagnosis of fetal congenital cystic adenomatoid malformation of the lung using three-dimensional ultrasound: comparison between the 20th and 21st centuries.

OBJECTIVE: Congenital cystic adenomatoid malformation of the lung (CCAML) is one of the most common lung lesions diagnosed prenatally. In order to compare the trends and improvements of prenatal diagnosis of CCAML, we herein retrospectively reviewed our cases of fetal CCAML detected by three-dimensional ultrasound (3-D US) between two centuries. MATERIALS AND METHODS: We reviewed our computer database of prenatal diagnosis of CCAML in National Cheng Kung University Hospital from October 1994 to November 2011. All of the fetuses were initially scanned by two-dimensional (2-D) US to locate the region-of-interest (ROI). Then, the 3-D probe was used to scan all of the ROI systematically and mechanically, and the images were stored in the laser discs for further 3-D visualization and reconstruction. To compare the characteristics at prenatal diagnosis of CCAML between the 20th and 21st centuries in our hospital, Chi-square tests were undertaken. A p value <0.05 was considered as statistically significant. RESULTS: In total, 58 fetuses with CCAML were depicted by 3-D US in utero (12 cases were diagnosed in the 20th century and 46 cases in the 21st century). The ranges of gestational age at prenatal diagnosis of CCAML by 3-D US in the 20th century were between 15 and 36 weeks (mean=24 weeks), and were between 16 and 31 weeks (mean=22 weeks) in the 21st century. Moreover, nine cases (75%) were diagnosed at the second trimester in the 20th century, whereas 44 cases (96%) were diagnosed at the second trimester in the 21st century. CONCLUSION: The advancement of 3-D US has remarkable advantages in adding novel visual depiction of a 3-D lesion of a 3-D fetus in 3-D US after reconstruction, and thus assists substantially in the prenatal diagnosis and genetic consultation of CCAML. Furthermore, the trend analysis in this series showed a significantly earlier gestational age at prenatal diagnosis of CCAML in the 21st century than that in the 20th century.

The enhancement of biofilm formation in Group B streptococcal isolates at vaginal pH.

Group B streptococcus (GBS) is a common asymptomatic colonizer in acidic vagina of pregnant women and can transmit to newborns, causing neonatal pneumonia and meningitis. Biofilm formation is often associated with bacterial colonization and pathogenesis. Little is known about GBS biofilm and the effect of environmental stimuli on their growth along with biofilm formation. The objective of this study was to investigate the survival and biofilm formation of GBS, isolated from pregnant women, in nutrient-limited medium under various pH conditions. Growth and survival experiments were determined by optical density and viable counts. Crystal violet staining, scanning electron microscopy, and atomic force microscopy (AFM) were used to analyze the capacity of biofilm production. Our results showed that GBS isolates proliferated with increasing pH with highest maximum specific growth rate (µmax) at pH 6.5, but survived at pH 4.5 for longer than 48 h. Biofilm formation of the 80 GBS isolates at pH 4.5 was significantly higher than at pH 7.0. This difference was confirmed by two other methods. The low elastic modulus obtained from samples at pH 4.5 by AFM revealed the softness of biofilm; in contrast, little or no biofilm was measured at pH 7.0. Under acidic pH, the capability of biofilm formation of serotypes III and V showed statistically significant difference from serotypes Ia and Ib. Our finding suggested that survival and enhanced biofilm formation at vaginal pH are potentially advantageous for GBS in colonizing vagina and increase the risk of vaginosis and neonatal infection.

Three-dimensional ultrasound in the prenatal diagnosis of osteogenesis imperfecta.

OBJECTIVE: Fetal osteogenesis imperfecta (OI) is a heterogeneous group of collagen disorders characterized by bone fragility, blue sclerae, deafness, and dentinogenesis imperfecta. Ultrasonography is acknowledged as a reliable diagnostic modality for the prenatal diagnosis of OI, especially type II. In the past, two-dimensional (2D) ultrasound (US) has been applied as the mainstay of prenatal diagnosis of OI. In this series, we report our work of detecting OI using three-dimensional (3D) US. MATERIAL AND METHODS: We reviewed our computer database of prenatal diagnosis of OI at the National Cheng Kung University Hospital from April 1996 to July 2010. All the cases were scanned by 2D and 3D US. In total, six cases of fetal OI were diagnosed. RESULTS: Compared with 2D US, 3D US can detect fetal OI precisely, and provide additional vivid illustration after various modes of reconstruction that 2D US cannot. CONCLUSION: In conclusion, 3D US may contribute significantly to the detection of OI in utero and provide a novel visual depiction of this defect after reconstruction. The technique may thus substantially assist in prenatal diagnosis as well as consultations for fetal OI.

Interleukin-1ß gene is not associated with preeclampsia in Taiwanese.

OBJECTIVE: To identify associations between the interleukin-1ß gene and preeclampsia in Taiwanese women. METHODS AND MATERIALS: We genotyped Taiwanese population (102 women with preeclampsia and 148 controls) for two polymorphisms of the interleukin-1ß gene (promoter region and Exon 5) by using polymerase chain reaction and restriction fragment length polymorphism analysis. The association between the genotype and disease was examined by Chi-square tests. RESULTS: We found no association between the two polymorphic sites of interleukin-1ß gene and preeclampsia. No significant differences were detected in genotype distributions and allele frequencies of the AvaI polymorphism at position -511 in the promoter region and the TaqI polymorphism at position +3953 within Exon 5. CONCLUSION: Our data do not support a role of the interleukin-1ß gene in the pathogenesis of preeclampsia in Taiwanese women.

Prenatal diagnosis of fetal multicystic dysplastic kidney in the era of three-dimensional ultrasound: 10-year experience.

OBJECTIVE: To demonstrate the usefulness of three-dimensional (3D) ultrasound in prenatal diagnosis of fetal multicystic dysplastic kidney (MCDK) disease. METHODS: In our previous study, we demonstrated that using 3D ultrasound in conjunction with traditional two-dimensional (2D) ultrasound can facilitate the diagnosis of MCDK. In this study, we followed all the MCDK cases diagnosed in our center in the recent decade (from 2002 to 2011) and compared the results with the data collected in the prior decade (from 1995 to 2002). RESULTS: Between 2002 and 2011, a total of 39 cases with fetal MCDK diagnosed by 2D and 3D ultrasound were retrospectively analyzed. The average gestational age when the diagnosis of MCDK was made was 23.6 weeks of gestation (95% confidence interval: 22.09-25.09). The Pearson chi-square test revealed a borderline nonsignificant difference statistically in the categorized gestational age at diagnosis (p = 0.052) as compared to the gestational age in the prior study. The average amniotic fluid index in fetuses with unilateral and bilateral MCDK was 16.76 ± 3.34 and 4.78 ± 5.82, respectively (p < 0.001). MCDK was not found to be associated with gestational age or chromosomal anomalies in our study. CONCLUSION: The surface-rendering mode of 3D ultrasound is very useful in prenatal diagnosis and counseling for MCDK.

Soft tissue volume of upper arm in predicting small-for-gestational-age fetuses using three-dimensional ultrasound.

PURPOSE: To assess the value of fetal soft tissue volume (STV) of the upper arm in predicting small-for-gestational-age (SGA) fetuses using three-dimensional (3D) ultrasound (US). METHODS: We used 3D US to test the accuracy of fetal STV of the upper arm measurement in predicting SGA in a prospective cross-sectional study. RESULTS: Fetal STV of the upper arm assessed by 3D US can differentiate SGA fetuses from appropriate-for-gestational-age (AGA) fetuses. Using the 5th percentile as the cutoff, the sensitivity of fetal upper arm STV in predicting SGA fetuses was 84.1%, specificity, 93.4%, positive predictive value, 71.1%, negative predictive value, 96.8%, and overall accuracy, 91.9%. In addition, the diagnostic accuracy of fetal arm STV was better than that of the biparietal diameter, abdominal circumference, and femur length. CONCLUSION: Fetal STV of upper arm assessment by 3D US is a novel method to predict SGA fetuses.

Hyaluronan substratum induces multidrug resistance in human mesenchymal stem cells via CD44 signaling.

Little information is available concerning multidrug resistance (MDR) in mesenchymal stem cells, although several studies have reported that MDR is associated with hyaluronan in neoplastic cells. We have evaluated whether a hyaluronan-coated surface modulates MDR in placenta-derived human mesenchymal stem cells (PDMSCs). We have found that PDMSCs cultured on a tissue-culture polystyrene surface coated with 30 microg/cm(2) hyaluronan are more resistant than control PDMSCs to doxorubicin. Inhibiting PI3K/Akt signaling has shown that the PI3K/Akt pathway modulates both P-glycoprotein activity and doxorubicin resistance. In addition, 10 microM verapamil dramatically suppresses the doxorubicin resistance induced by the hyaluronan-coated surface, indicating that P-glycoprotein activity is necessary for MDR. We have further found that PDMSCs treated with CD44 small interfering RNA (siRNA) and grown on a polystyrene surface coated with 30 microg/cm(2) hyaluronan have fewer P-glycoprotein(+) cells and lower CD44 expression levels (less than 60% in both cases) than PDMSCs not treated with CD44 siRNA and grown on the hyaluronan-coated surface. Moreover, treatment with CD44 siRNA suppresses the hyaluronan-substratum-induced doxorubicin resistance. We conclude that a hyaluronan substratum induces MDR in PDMSCs through CD44 signaling.