Induction of zonal-specific cellular morphology and matrix synthesis for biomimetic cartilage regeneration using hybrid scaffolds.

Cartilage is anisotropic in nature and organized into distinct zones. Our goal was to develop zonal-specific three-dimensional hybrid scaffolds which could induce the generation of zonal-specific cellular morphology and extracellular matrix (ECM) composition. The superficial and middle zones comprised two layers of hyaluronic acid (HA) hydrogel which enveloped specifically orientated or randomly arranged polylactic acid nanofibre meshes. The deep zone comprised a HA hydrogel with multiple vertical channels. Primary bovine chondrocytes were seeded into the individual zonal scaffolds, cultured for 14 days and then the ECM was analysed. The aligned nanofibre mesh used in the superficial zone induced an elongated cell morphology, lower glycosaminoglycan (GAG) and collagen II production, and higher cell proliferation and collagen I production than the cells in the middle zone scaffold. Within the middle zone scaffold, which comprised a randomly orientated nanofibre mesh, the cells were clustered and expressed more collagen II. The deep zone scaffold induced the highest GAG production, the lowest cell proliferation and the lowest collagen I expression of the three zones. Assembling the three zones and stabilizing the arrangement with a HA hydrogel generated aligned, randomly aggregated and columnar cells in the superficial, middle and deep zones. This study presents a method to induce zonal-specific chondrocyte morphology and ECM production.

Human tRNA-Derived Small RNAs Modulate Host-Oral Microbial Interactions.

Coevolution of the human host and its associated microbiota has led to sophisticated interactions to maintain a delicate homeostasis. Emerging evidence suggests that in addition to small molecules, peptides, and proteins, small regulatory noncoding RNAs (sRNAs) might play an important role in cross-domain interactions. In this study, we revealed the presence of diverse host transfer RNA-derived small RNAs (tsRNAs) among human salivary sRNAs. We selected 2 tsRNAs (tsRNA-000794 and tsRNA-020498) for further study based on their high sequence similarity to specific tRNAs from a group of Gram-negative oral bacteria, including Fusobacterium nucleatum, a key oral commensal and opportunistic pathogen. We showed that the presence of F. nucleatum triggers exosome-mediated release of tsRNA-000794 and tsRNA-020498 by human normal oral keratinocyte cells. Furthermore, both tsRNA candidates exerted a growth inhibition effect on F. nucleatum, likely through interference with bacterial protein biosynthesis, but did not affect the growth of Streptococcus mitis, a health-associated oral Gram-positive bacterium whose genome does not carry sequences bearing high similarity to either tsRNA. Our data provide the first line of evidence for the modulatory role of host-derived tsRNAs in the microbial-host interaction.

Microbiome Associated with Severe Caries in Canadian First Nations Children.

Young Indigenous children in North America suffer from a higher degree of severe early childhood caries (S-ECC) than the general population, leading to speculation that the etiology and characteristics of the disease may be distinct in this population. To address this knowledge gap, we conducted the first microbiome analysis of an Indigenous population using modern molecular techniques. We investigated the caries-associated microbiome among Canadian First Nations children with S-ECC. Thirty First Nations children <72 mo of age with S-ECC and 20 caries-free children were recruited in Winnipeg, Canada. Parents or caregivers completed a questionnaire on general and dental health, diet, and demographics. The plaque microbiome was investigated by sequencing the 16S rRNA gene. Sequences were clustered into operational taxonomic units and taxonomy assigned via the Human Oral Microbiome Database, then analyzed at the community level with alpha and beta diversity measures. Compared with those who were caries free, children with S-ECC came from households with lower income; they were more likely to live in First Nations communities and were more likely to be bottle-fed; and they were weaned from the bottle at a later age. The microbial communities of the S-ECC and caries-free groups did not differ in terms of species richness or phylogenetic diversity. Beta diversity analysis showed that the samples significantly clustered into groups based on caries status. Twenty-eight species-level operational taxonomic units were significantly different between the groups, including Veillonella HOT 780 and Porphyromonas HOT 284, which were 4.6- and 9-fold higher, respectively, in the S-ECC group, and Streptococcus gordonii and Streptococcus sanguinis, which were 5- and 2-fold higher, respectively, in the caries-free group. Extremely high levels of Streptococcus mutans were detected in the S-ECC group. Overall, First Nations children with S-ECC have a significantly different plaque microbiome than their caries-free counterparts, with the S-ECC group containing higher levels of known cariogenic organisms.

Arginine Improves pH Homeostasis via Metabolism and Microbiome Modulation.

Dental caries can be described as a dysbiosis of the oral microbial community, in which acidogenic, aciduric, and acid-adapted bacterial species promote a pathogenic environment, leading to demineralization. Alkali generation by oral microbes, specifically via arginine catabolic pathways, is an essential factor in maintaining plaque pH homeostasis. There is evidence that the use of arginine in dentifrices helps protect against caries. The aim of the current study was to investigate the mechanistic and ecological effect of arginine treatment on the oral microbiome and its regulation of pH dynamics, using an in vitro multispecies oral biofilm model that was previously shown to be highly reflective of the in vivo oral microbiome. Pooled saliva from 6 healthy subjects was used to generate overnight biofilms, reflecting early stages of biofilm maturation. First, we investigated the uptake of arginine by the cells of the biofilm as well as the metabolites generated. We next explored the effect of arginine on pH dynamics by pretreating biofilms with 75 mM arginine, followed by the addition of sucrose (15 mM) after 0, 6, 20, or 48 h. pH was measured at each time point and biofilms were collected for 16S sequencing and targeted arginine quantification, and supernatants were prepared for metabolomic analysis. Treatment with only sucrose led to a sustained pH drop from 7 to 4.5, while biofilms treated with sucrose after 6, 20, or 48 h of preincubation with arginine exhibited a recovery to higher pH. Arginine was detected within the cells of the biofilms, indicating active uptake, and arginine catabolites citrulline, ornithine, and putrescine were detected in supernatants, indicating active metabolism. Sequencing analysis revealed a shift in the microbial community structure in arginine-treated biofilms as well as increased species diversity. Overall, we show that arginine improved pH homeostasis through a remodeling of the oral microbial community.

Bilateral retinal microglial response to unilateral optic nerve transection in rats.

When retinal ganglion cells undergo apoptosis after optic nerve (ON) injury, microglial cells proliferate and promptly clear the degenerated debris in the ipsilateral retina. However, microglial changes in the contralateral retina have not been fully elucidated. This study characterized the long-term bilateral retinal microglial responses after unilateral ON transection. We analyzed the time course of proliferation and morphology changes of microglial cells, between 3 days and 12 weeks post ON transection, of undisturbed and reactive microglia in bilateral retinas of adult Fischer rats with unilateral ON transection. Microglia in retinas without ON transection were distributed homogeneously and possessed a highly ramified morphology, as judged by immunohistochemistry for ionized calcium-binding adapter molecule 1 (Iba1). After ON transection, microglia density in the ipsilateral retina increased gradually from 3 days to 2 weeks, and decreased from 3 weeks to 12 weeks, along with dramatic inverted alteration of process branch points of microglia in the ganglion cell layer (GCL). Transformation of ramified microglia into ameboid-like macrophages with few branching processes was observed in the ipsilateral retina from 1 week to 3 weeks. Though an increase in microglial density was weak in the contralateral retina and could only be statistically detected in the central retina, the morphological alteration over time was obvious and similar to that of the ipsilateral retina. In the inner plexiform layer (IPL), cell density and morphological changes of microglia in both the ipsilateral and contralateral retina were not prominent. These findings indicates that, though proliferation of microglial cells is weak in the contralateral retina after unilateral ON transection, conspicuous alterations in microglial morphology occur bilaterally. These suggest that using the contralateral retina as a control in studies of retinal degeneration should be considered with caution.

Cellular Components Mediating Coadherence of Candida albicans and Fusobacterium nucleatum.

Candida albicans is an opportunistic fungal pathogen found as part of the normal oral flora. It can be coisolated with Fusobacterium nucleatum, an opportunistic bacterial pathogen, from oral disease sites, such as those involved in refractory periodontitis and pulp necrosis. The physical coadherence between these 2 clinically important microbes has been well documented and suggested to play a role in facilitating their oral colonization and colocalization and contributing to polymicrobial pathogenesis. Previous studies indicated that the physical interaction between C. albicans and F. nucleatum was mediated by the carbohydrate components on the surface of C. albicans and the protein components on the Fusobaterium cell surface. However, the identities of the components involved still remain elusive. This study was aimed at identifying the genetic determinants involved in coaggregation between the 2 species. By screening a C. albicans SN152 mutant library and a panel of F. nucleatum 23726 outer membrane protein mutants, we identified FLO9, which encodes a putative adhesin-like cell wall mannoprotein of C. albicans and radD, an arginine-inhibitable adhesin-encoding gene in F. nucleatum that is involved in interspecies coadherence. Consistent with these findings, we demonstrated that the strong coaggregation between wild-type F. nucleatum 23726 and C. albicans SN152 in an in vitro assay could be greatly inhibited by arginine and mannose. Our study also suggested a complex multifaceted mechanism underlying physical interaction between C. albicans and F. nucleatum and for the first time revealed the identity of major genetic components involved in mediating the coaggregation. These observations provide useful knowledge for developing new targeted treatments for disrupting interactions between these 2 clinically relevant pathogens.

Bio-inspired aquatic robotics by untethered piezohydroelastic actuation.

This paper investigates fish-like aquatic robotics using flexible bimorphs made of macro-fiber composite (MFC) piezoelectric laminates for carangiform locomotion. In addition to noiseless and efficient actuation over a range of frequencies, geometric scalability, and simple design, bimorph propulsors made of MFCs offer a balance between the actuation force and velocity response for performance enhancement in bio-inspired swimming. The experimental component of the presented work focuses on the characterization of an elastically constrained MFC bimorph propulsor for thrust generation in quiescent water as well as the development of a robotic fish prototype combining a microcontroller and a printed-circuit-board amplifier to generate high actuation voltage for untethered locomotion. From the theoretical standpoint, a distributed-parameter electroelastic model including the hydrodynamic effects and actuator dynamics is coupled with the elongated-body theory for predicting the mean thrust in quiescent water. In-air and underwater experiments are performed to verify the incorporation of hydrodynamic effects in the linear actuation regime. For electroelastically nonlinear actuation levels, experimentally obtained underwater vibration response is coupled with the elongated-body theory to predict the thrust output. The measured mean thrust levels in quiescent water (on the order of ∼10 mN) compare favorably with thrust levels of biological fish. An untethered robotic fish prototype that employs a single bimorph fin (caudal fin) for straight swimming and turning motions is developed and tested in free locomotion. A swimming speed of 0.3 body-length/second (7.5 cm s⁻¹ swimming speed for 24.3 cm body length) is achieved at 5 Hz for a non-optimized main body-propulsor bimorph combination under a moderate actuation voltage level.

IDH2 mutations are frequent in Chinese patients with acute myeloid leukemia and associated with NPM1 mutations and FAB-M2 subtype.

INTRODUCTION: Gene mutations play an important role in acute myeloid leukemia (AML) pathogenesis. Several genes have been identified in AML, such as FLT3, KIT, NPM1, and JAK2. This study investigated the frequency of novel mutations in IDH1 (amino acid R132) and IDH2 (R140 and R172) and analyzed their impact on disease biology and interaction with other mutations in Chinese patients with de novo AML. METHODS: A total of 195 patients were screened for mutations in the IDH1, IDH2, JAK2 V617F, NPM1, FLT3, and KIT genes, using polymerase chain reaction (PCR)-based and direct sequencing assays. RESULTS: IDH mutations occurred at a considerable frequency of 15.89% in Chinese AML cases; IDH2 R140Q was the most frequent genetic alteration and was associated with older age, normal karyotype, and French-American-British classification M2 at diagnosis. There was a strong association of IDH2 mutation with NPM1 mutations and a trend with FLT3-internal-tandem duplication. CONCLUSION: IDH mutations may be a novel genetic marker in cytogenetically normal AML and may cooperate in leukemogenesis.

Randomized multicenter clinical trial of myofascial physical therapy in women with interstitial cystitis/painful bladder syndrome and pelvic floor tenderness.

PURPOSE: We determined the efficacy and safety of pelvic floor myofascial physical therapy compared to global therapeutic massage in women with newly symptomatic interstitial cystitis/painful bladder syndrome. MATERIALS AND METHODS: A randomized controlled trial of 10 scheduled treatments of myofascial physical therapy vs global therapeutic massage was performed at 11 clinical centers in North America. We recruited women with interstitial cystitis/painful bladder syndrome with demonstrable pelvic floor tenderness on physical examination and a limitation of no more than 3 years' symptom duration. The primary outcome was the proportion of responders defined as moderately improved or markedly improved in overall symptoms compared to baseline on a 7-point global response assessment scale. Secondary outcomes included ratings for pain, urgency and frequency, the O'Leary-Sant IC Symptom and Problem Index, and reports of adverse events. We compared response rates between treatment arms using the exact conditional version of the Mantel-Haenszel test to control for clustering by clinical center. For secondary efficacy outcomes cross-sectional descriptive statistics and changes from baseline were calculated. RESULTS: A total of 81 women randomized to the 2 treatment groups had similar symptoms at baseline. The global response assessment response rate was 26% in the global therapeutic massage group and 59% in the myofascial physical therapy group (p=0.0012). Pain, urgency and frequency ratings, and O'Leary-Sant IC Symptom and Problem Index decreased in both groups during followup, and were not significantly different between the groups. Pain was the most common adverse event, occurring at similar rates in both groups. No serious adverse events were reported. CONCLUSIONS: A significantly higher proportion of women with interstitial cystitis/painful bladder syndrome responded to treatment with myofascial physical therapy than to global therapeutic massage. Myofascial physical therapy may be a beneficial therapy in women with this syndrome.

Comparative impact on prostanoid biosynthesis of celecoxib and the novel nonsteroidal anti-inflammatory drug CG100649.

Nonsteroidal anti-inflammatory drugs (NSAIDs) elevate cardiovascular risk by disrupting cyclooxygenase-2 (COX-2)-dependent biosynthesis of prostacyclin (PGI(2)). CG100649 is a novel NSAID proposed to inhibit both COX-2 and carbonic anhydrase (CA)-I/-II. We compared its impact on prostanoid biosynthesis with that of celecoxib, an NSAID purposefully designed to selectively inhibit COX-2. In a controlled, double-blind randomized trial, single oral doses of 2 or 8 mg CG100649, 200 mg celecoxib, or placebo were well tolerated by healthy volunteers (n = 23). Both CG100649 and celecoxib had the effect of depressing urinary excretion of 2,3-dinor-6-keto-PGF(1α) (PGI-M); the effect of CG100649 was dose-dependent and more sustained (up to 240 h after the dose) than that of celecoxib. Neither CG100649 nor celecoxib significantly inhibited COX-1-dependent prostanoid formation. CA inhibition was not detected after administration of CG100649, despite its partitioning asymmetrically into erythrocytes. CG100649 and celecoxib are both relatively selective inhibitors of COX-2, but they differ in duration of action. Whether they have similar impact on cardiovascular events remains to be determined.

Multiple roles of the p75 neurotrophin receptor in the nervous system.

The p75 neurotrophin receptor (p75NTR) is a transmembrane protein that binds nerve growth factor (NGF) and has multiple functions in the nervous system where it is expressed widely during the developmental stages of life, although expression decreases dramatically by adulthood. Expression of p75NTR can increase in pathological states related to neural cell death. p75NTR is a member of the tumour necrosis factor (TNF) receptor family and it consists of intracellular, transmembrane and extracellular domains which are different from other TNF receptors. Either by interacting with tropomyosin receptor kinase (Trk) receptors or via the independent binding of neurotrophin, p75NTR can induce neurite outgrowth and cellular survival or cell apoptosis through several complicated signal transduction pathways. Most of these signalling pathways remain to be elucidated. By interacting with different cellular factors, p75NTR can induce neuron growth cone collapse or regrowth. p75NTR is also expressed in a variety of glial populations. The many functions of p75NTR require further study.

LLL-3 inhibits STAT3 activity, suppresses glioblastoma cell growth and prolongs survival in a mouse glioblastoma model.

Persistent activation of the signal transducer and activator of transcription 3 (STAT3) signalling has been linked to oncogenesis and the development of chemotherapy resistance in glioblastoma and other cancers. Inhibition of the STAT3 pathway thus represents an attractive therapeutic approach for cancer. In this study, we investigated the inhibitory effects of a small molecule compound known as LLL-3, which is a structural analogue of the earlier reported STAT3 inhibitor, STA-21, on the cell viability of human glioblastoma cells, U87, U373, and U251 expressing constitutively activated STAT3. We also investigated the inhibitory effects of LLL-3 on U87 glioblastoma cell growth in a mouse tumour model as well as the impact it had on the survival time of the treated mice. We observed that LLL-3 inhibited STAT3-dependent transcriptional and DNA binding activities. LLL-3 also inhibited viability of U87, U373, and U251 glioblastoma cells as well as induced apoptosis of these glioblastoma cell lines as evidenced by increased poly (ADP-ribose) polymerase (PARP) and caspase-3 cleavages. Furthermore, the U87 glioblastoma tumour-bearing mice treated with LLL-3 exhibited prolonged survival relative to vehicle-treated mice (28.5 vs 16 days) and had smaller intracranial tumours and no evidence of contralateral invasion. These results suggest that LLL-3 may be a potential therapeutic agent in the treatment of glioblastoma with constitutive STAT3 activation.

Controllable coherent population transfers in superconducting qubits for quantum computing.

We propose an approach to coherently transfer populations between selected quantum states in one- and two-qubit systems by using controllable Stark-chirped rapid adiabatic passages. These evolution-time insensitive transfers, assisted by easily implementable single-qubit phase-shift operations, could serve as elementary logic gates for quantum computing. Specifically, this proposal could be conveniently demonstrated with existing Josephson phase qubits. Our proposal can find an immediate application in the readout of these qubits. Indeed, the broken parity symmetries of the bound states in these artificial atoms provide an efficient approach to design the required adiabatic pulses.

Differential roles of phosphatidylinositol 3-kinase/akt pathway in retinal ganglion cell survival in rats with or without acute ocular hypertension.

Intraocular pressure (IOP) elevation has often been used as an experimental model to study mechanisms underlying retinal ganglion cell (RGC) death associated with ocular ischemic injury and glaucoma. The aim of the present study, using both in vitro and in vivo approaches, was to investigate the role of phosphatidylinositol 3-kinase (PI3K)/akt pathway in RGC viability in normal rats and rats following transient IOP elevation. For in vivo studies, pathway inhibitors were administered intravitreally on days 3, 9, and 15 post-2-h IOP elevation at 110 mm Hg. Toward the end of the 3-week examination period, the fluorescent dye Fluorogold was used to retrogradely label surviving RGCs. In order to examine the role of macrophages that were recruited into the eye following the pathway inhibition, clodronate liposomes were used to deplete phagocytic cells in the eye. PI3K/akt pathway activity and location in the retina were examined using Western blot and immunohistochemistry, respectively. Here we showed that PI3K/akt inhibitors 2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride (LY294002) and KY12420 at low concentrations (2 microM or 20 microM) did not influence RGC survival but caused RGC loss at high concentration (200 muM) in retinal explants derived from intact rats. In contrast, both LY294002 and KY12420 at 20 microM led to RGC loss in retinal explants derived from IOP-elevated eyes. A detrimental action of phagocytic cells on RGC survival was also seen in these retinas. In vivo results confirmed the detrimental actions of PI3K/akt inhibition and macrophages on RGC survival in IOP-elevated, but not intact eyes even with high concentration of LY294002. Low level of PI3K/akt activity was detected in the ganglion cell layer (GCL) in intact retina. Acute IOP elevation activated PI3K/akt pathway in the inner nuclear layer and GCL including RGCs. This study thus demonstrates that PI3K/akt pathway mediates RGC survival after IOP elevation but not under normal condition.

PDK-1/AKT pathway as a novel therapeutic target in rhabdomyosarcoma cells using OSU-03012 compound.

Rhabdomyosarcoma (RMS) is the most common paediatric soft-tissue sarcoma including two major subtypes, alveolar rhabdomyosarcoma (ARMS) and embryonal rhabdomyosarcoma (ERMS). Increasing evidence suggests that oncogenesis of RMS involves multiple stages of signalling protein dysregulation which may include prolonged activation of serine/threonine kinases such as phosphoinositide-dependent kinase-1 (PDK-1) and AKT. To date, whether PDK-1/AKT pathway is activated in RMS is unknown. This study was to examine phosphorylation status of AKT and to evaluate a novel small molecular inhibitor, OSU-03012 targeting PDK-1 in RMS. We examined phosphorylation levels of AKT using ARMS and ERMS tissue microarray and immunohistochemistry staining. Our results showed phospho-AKT(Thr308) level is elevated 42 and 35% in ARMS and ERMS, respectively. Phospho-AKT(Ser473) level is also increased 43% in ARMS and 55% in ERMS. Furthermore, we showed that OSU-03012 inhibits cell viability and induces apoptosis in ARMS and ERMS cell lines (RH30, SMS-CTR), which express elevated phospho-AKT levels. Normal cells are much less sensitive to OSU-03012 and in which no detectable apoptosis was observed. This study showed, for the first time, that PDK-1/AKT pathway is activated in RMS and may play an important role in survival of RMS. PDK-1/AKT pathway may be an attractive therapeutic target for cancer intervention in RMS using OSU-03012.

Antibacterial and antifungal efficacy of surface functionalized polymeric beads in repeated applications.

A simple method was developed to prepare polymeric microbeads with antibacterial and antifungal properties. The microbeads of approximately spherical shape and narrow size distribution were prepared from a mixture of poly (4-vinyl pyridine) (P4VP) and poly (vinylidene fluoride) (PVDF) by a phase inversion technique and subsequently derivatized with alkyl bromides having 4-10 carbon atoms. The quaternization of the pyridine groups into pyridinium groups confers the surface with highly effective and long-lasting antibacterial and antifungal properties, as shown by the effect on Escherichia coli and Aspergillus niger. Upon contact with the N-alkylated beads, the bacteria and fungal spores are lysed and intracellular constituents leach out into the medium. The efficacy of the alkyl chains in disrupting the cell membrane was investigated. The stability of the functional group and microbiocidal effectiveness of the microbeads in repeated applications was also assessed.

Antibacterial activity of cloth functionalized with N-alkylated poly(4-vinylpyridine).

A simple technique was developed to functionalize the surface of commercial cotton cloth with bactericidal polycationic groups. The cloth was first graft copolymerized with 4-vinylpyridine (4VP) and subsequently derivatized with hexylbromide via the quaternization of the grafted pyridine groups into pyridinium groups. X-ray photoelectron spectroscopy (XPS) was used to characterize the cotton cloth after each surface functionalization step. The pyridinium groups introduced on the substrate surface exhibited a high bacterial killing efficiency as shown by airborne and waterborne Escherichia coli (E. coli) assays, as well as a test to measure the number of viable E. coli cells in suspension upon contact with the cloth. This antibacterial property is preserved even after the cloth is subjected to prolonged weathering under UV irradiation and water spray. Wild-type bacteria were also cultured from soil and used to verify the antibacterial potential of the functionalized cloth. The inhibition of biofilm formation on the functionalized cloth was observed even with the inoculation of high concentrations of microorganisms. The success of the present surface functionalization technique as applied to cloth is a promising development, as since cloth is a common material having wide applications.

[Determination of salicylic acid, resorcinol and chloramphenicol in junlieping tincture by reversed-phase high performance liquid chromatography].

A reversed-phase high performance liquid chromatographic method was developed for the quantitative determination of salicylic acid, resorcinol and chloramphenicol in Junlieping tincture. The operation was carried out on an ODS column with the mobile phase of a mixture of acetonitrile-methanol-water(25:10:65, volume proportion). The eluate was detected at 278 nm wavelength. This method is simple, fast and can be used for the quality control of Junlieping tincture.

A short review of the previous and current epidemiological situation of schistosomiasis in China.

Schistosomiasis japonica has long been endemic in the People's Republic of China. In the 1950s, the number of the infected persons was estimated at 10 million; infected snail habitats were estimated at more than 14 billion square metres and infected cattle at 1.2 million. After schistosomiasis control measures were carried out, it was a great success. According to the survey of 1989, infected persons were estimated at 0.95 million; infected snails at 3.47 billion and infected cattle at about 0.1 million. These results compared with those of the 1950s show big reductions in prevalence rates 90.5%, 75.2% and 91.6%, respectively. At present, the disease is a threat in the marshland and lake regions and the high mountainous regions. To maintain the success achieved in effective control and to bring the yet endemic marshland and lake regions and mountainous areas under control are hard and long-term tasks confronting the People's Republic of China.

[The disturbance of calcium homeostasis in vascular smooth muscle proliferation after balloon denudation].

OBJECTIVE: To investigate the changes of calcium homeostasis in smooth muscle cells (VSMC) and the role of disturbance of calcium homeostasis in VSMC proliferation after balloon denudation. METHODS: Assay of the cellular incorporation of 3H-leucine and measurement of 45Ca transport were done on the model of balloon-denuded aorta in rat. RESULTS: Endothelial injury induced VSMC proliferation, intimal thickening. After balloon denudation, VSMC calcium influx increased (3 days after balloon denudation, 3.28 +/- 0.14 vs 4.12 +/- 0.28, P < 0.05; 10 days 3.31 +/- 0.09 vs 4.09 +/- 0.21 nmol/10(6) cells, P < 0.01), and calcium efflux decreased, and calcium content increased (3 days after balloon denudation 695 +/- 33 vs 995 +/- 54, P < 0.01; 10 days 709 +/- 32 vs 1022 +/- 94 nmol/10(6) cell, P < 0.01). SR and mitochondria calcium uptake increased. Calcium antagonist, verapamil not only regulated the disturbance of calcium homeostasis, but also inhibited endothelium injury-induced VSMC proliferation. CONCLUSION: The disturbance of calcium homeostasis is probably one of the underlying mechanisms of VSMC proliferation induced by balloon denudation.