Preservation of human heart valves for replacement in children with heart valve disease: past, present and future.

Valvular heart disease affects 30% of the new-borns with congenital heart disease. Valve replacement of semilunar valves by mechanical, bioprosthetic or donor allograft valves is the main treatment approach. However, none of the replacements provides a viable valve that can grow and/or adapt with the growth of the child leading to re-operation throughout life. In this study, we review the impact of donor valve preservation on moving towards a more viable valve alternative for valve replacements in children or young adults.

Efficacy of Proximal Resin Infiltration on Caries Inhibition: Results from a 3-Year Randomized Controlled Clinical Trial.

This study reports 3-y outcomes of a split-mouth randomized clinical trial. Resin infiltration's capacity to arrest caries lesion progression in noncavitated proximal lesions is affirmed. Forty-two consented young adults, blinded to tooth surface allocation, were treated with resin infiltration on 1 randomly selected surface and concurrently experienced a mock infiltration procedure on another. Both treatments were provided as an adjunct to the currently accepted standard-of-care regimen (periodic prophylaxis and serial fluoride varnish applications) appropriate for the management of high caries risk. Challenging periods of low oral hygiene compliance were expected. The primary outcome measure was 3-y radiographic lesion progression. Blinded investigators evaluated each study surface for lesion progression with a series of images obtained at intervals over the 3-y course of study. Proportions of progressing lesions were compared with McNemar's test. Twenty-nine noncavitated lesion pairs in permanent posterior teeth demonstrating caries penetrating into inner enamel or outer dentin were included in the analyses. No adverse events were reported. Radiographic progression was recorded in 4 of 29 infiltrated lesions (14%) and 14 of 29 control lesions (48%, P < .003). Adjunct resin infiltration demonstrated a high 3-y efficacy of 71% (relative risk reduction). The prevented fraction was 86% for infiltration versus 52% for controls. Resin infiltration was 100% successful in arresting caries progression in inner enamel lesions (E2) and 64% in outer dentin lesions (D1). Supplementary microinvasive resin infiltration is significantly more efficacious in reducing proximal lesion progression than management by standard noninvasive therapy alone. Long-term results may shed light on whether this represents the arrest or delay of the caries disease process (ClinicalTrials.gov NCT01584024).

Monitoring of sound and carious surfaces under sealants over 44 months.

Although there is strong evidence for the effectiveness of sealants, one major barrier in sealant utilization is the concern of sealing over active caries lesions. This study evaluated detection and monitoring of caries lesions through a clear sealant over 44 mo. Sixty-four 7- to 10-year-old children with at least 2 permanent molars with International Caries Detection and Assessment System (ICDAS) scores 0-4 (and caries less than halfway through the dentin, radiographically) were examined with ICDAS, DIAGNOdent, and quantitative light-induced fluorescence (QLF) before sealant placement and 1, 12, 24, and 44 mo (except QLF) after. Bitewing radiographs were taken yearly. DIAGNOdent and QLF were able to distinguish between baseline ICDAS before and after sealant placement. There was no significant evidence of ICDAS progression at 12 mo, but there was small evidence of minor increases at 24 and 44 mo (14% and 14%, respectively) with only 2% ICDAS ≥ 5. Additionally, there was little evidence of radiographic progression (at 12 mo = 1%, 24 mo = 3%, and 44 mo = 9%). Sealant retention rates were excellent at 12 mo = 89%, 24 mo = 78%, and 44 mo = 70%. The small risk of sealant repair increased significantly as baseline ICDAS, DIAGNOdent, and QLF values increased. However, regardless of lesion severity, sealants were 100% effective at 12 mo and 98% effective over 44 mo in managing occlusal surfaces at ICDAS 0-4 (i.e., only 4 of 228 teeth progressed to ICDAS ≥ 5 associated with sealants in need of repair and none to halfway or more through the dentin, radiographically). This study suggests that occlusal surfaces without frank cavitation (ICDAS 0-4) that are sealed with a clear sealant can be monitored with ICDAS, QLF, or DIAGNOdent, which may aid in predicting the need for sealant repair.

Quantification of continual anthropogenic pollutants released in swimming pools.

Disinfection in swimming pools is often performed by chlorination, However, anthropogenic pollutants from swimmers will react with chlorine and form disinfection by-products (DBPs). DBPs are unwanted from a health point of view, because some are irritating, while others might be carcinogenic. The reduction of anthropogenic pollutants will lead to a reduction in DBPs. This paper investigates the continual release of anthropogenic pollutants by means of controlled sweat experiments in a pool tank during laboratory time-series experiments (LTS experiments) and also during on-site experiments (OS experiments) in a swimming pool. The sweat released during the OS and LTS experiments was very similar. The sweat rate found was 0.1-0.2 L/m(2)/h at water temperatures below 29 °C and increased linearly with increasing water temperatures to 0.8 L/m(2)/h at 35 °C. The continual anthropogenic pollutant release (CAPR) not only consisted of sweat, particles (mainly skin fragments and hair) and micro-organisms, but also sebum (skin lipids) has to be considered. The release of most components can be explained by the composition of sweat. The average release during 30 min of exercise is 250 mg/bather non-purgeable organic carbon (NPOC), 77.3 mg/bather total nitrogen (TN), 37.1 mg/bather urea and 10.1 mg/bather ammonium. The release of NPOC cannot be explained by the composition of sweat and is most probably a result of sebum release. The average release of other components was 1.31 × 10(9) # particles/bather (2-50 µm), 5.2 µg/bather intracellular adenosine triphosphate (cATP) and 9.3 × 10(6) intact cell count/bather (iCC). The pool water temperature was the main parameter to restrain the CAPR. This study showed that a significant amount of the total anthropogenic pollutants release is due to unhygienic behaviour of bathers.

Clinical reduction of S. mutans in pre-school children using a novel liquorice root extract lollipop: a pilot study.

AIM: To determine the clinical effect of a simple herbal caries-prevention protocol aimed at reduction of Streptococcus mutans (SM) in young children in a pre-school setting. STUDY DESIGN: Proof-of-principle pilot study. METHODS: To prove the concept this pilot study delivered a clinical intervention using sugar-free lollipops containing liquorice root extract. Regimen: Supervised herbal lollipops, twice daily for 3 weeks. Species-specific monoclonal antibody testing of saliva provided SM counts. Children were grouped in high, medium and low caries-risk using baseline SM-levels as risk-indicator. Bacterial numbers at baseline, during intervention, and for 9 weeks post-intervention were compared. STATISTICS: SM levels were analysed using GEE modelling. RESULTS: High-risk children showed the steepest early decrease in mean log-SM (P<.001). At end of a follow-up period, the log-SM decrease moved the high-risk group down to moderate-risk level. High-risk children showed a decrease in fitted mean SM% not seen in other groups (P<.001). The decrease reached a nadir around 22-days post-intervention. Twice-daily use of herbal lollipop significantly reduced both number and relative percent of SM in high-risk children. SM numbers were reduced for 22 days after the last lollipop, stabilized and then began to rebound. CONCLUSION: A potential for simple effective caries-prevention for high-risk children has been demonstrated. Encouraging results warrant randomised clinical trials (RCT) of liquorice root in herbal lollipops or alternative modes of delivery.

In vivo acid etching effect on bacteria within caries-affected dentin.

Acid etching procedures may disrupt residual bacteria and contribute to the success of incomplete caries removal followed by adhesive restoration. This study evaluated the in vivo effect of acid etching on cariogenic bacterial activity within affected dentin after minimally invasive treatment of caries lesions. Twenty-eight carious permanent teeth received standardized selective caries removal and random acid etch treatment (E) or not (NE) prior to adhesive restoration. Baseline and 3-month dentin biopsies were collected. The number of bacteria and activity of total bacterial cells and Streptococcus mutans were determined by quantitative PCR and RT-PCR. No statistically significant differences were observed in total bacterial number and activity between E and NE treatments (p > 0.3008). For NE, however, the residual S. mutans bacterial cells were reduced (p = 0.0027), while the activity per cell was significantly increased (p = 0.0010) after reentry at 3 months after restoration. This effect was not observed in group E. Although no significant differences were found between groups, this study suggests that acid etching of affected dentin prior to adhesive restoration may directly or indirectly have an inhibitive effect on the activity of residual cariogenic bacteria. Further research is required to investigate this potential effect.

In vivo dentin microhardness beneath a calcium-phosphate cement.

A minimally invasive caries-removal technique preserves potentially repairable, caries-affected dentin. Mineral-releasing cements may promote remineralization of soft residual dentin. This study evaluated the in vivo remineralization capacity of resin-based calcium-phosphate cement (Ca-PO(4)) used for indirect pulp-capping. Permanent carious and sound teeth indicated for extraction were excavated and restored either with or without the Ca-PO(4) base (control), followed by adhesive restoration. Study teeth were extracted after 3 months, followed by sectioning and in vitro microhardness analysis of the cavity floor to 115-microm depth. Caries-affected dentin that received acid conditioning prior to Ca-PO(4) basing showed significantly increased Knoop hardness near the cavity floor. The non-etched group presented results similar to those of the non-treated group. Acid etching prior to cement application increased microhardness of residual dentin near the interface after 3 months in situ.

In vivo dentin remineralization by calcium-phosphate cement.

Minimally invasive caries-removal procedures remove only caries-infected dentin and preserve caries-affected dentin that becomes remineralized. Dental cements containing calcium phosphate promote remineralization. This study evaluated the in vivo remineralization capacity of resin-based calcium-phosphate cement (Ca-P) used for indirect pulp-capping. Carious and sound teeth indicated for extraction were randomly restored with the Ca-P base or without base (control), followed by adhesive restoration. Study teeth were extracted after three months, followed by elemental analysis of the cavity floor. Mineral content of affected or sound dentin at the cavity floor was quantified by electron probe micro-analysis to 100-mum depth. After three months, caries-affected dentin underneath the Ca-P base showed significantly increased calcium and phosphorus content to a depth of 30 mum. Mineral content of treated caries-affected dentin was in the range of healthy dentin, revealing the capacity of Ca-P base to promote remineralization of caries-affected dentin.

Effect of bur type and conditioning on the surface and interface of dentine.

The purpose of this in vitro study was to evaluate the surface and resin-dentine interface characteristics of permanent tooth dentine cut with diamond or carbide burs and treated with phosphoric acid (PA) or an acidic conditioner. Labial surfaces of permanent incisors were prepared into dentine with high-speed carbide or diamond burs and divided into two halves. Phosphoric acid 36% was applied on one half and non-rinse conditioner (NRC) was applied on the other half. Ten randomly selected scanning electron microscopy (SEM) fields from each specimen (n = 15) were evaluated. Occlusal surfaces of third molars were divided in two halves for evaluation of the resin-dentine interface. The halves were randomly assigned to one of each conditioner and restored with Prime & Bond NT/Spectrum. Ten specimens were analysed by SEM to evaluate hybrid layer formation and interfacial seal. We observed that surfaces prepared with carbide bur presented less residual smear plugs (P < 0.05) than surfaces prepared with diamond burs. Surfaces conditioned with NRC, which is a smear layer modifier, presented more residual smear plugs than surfaces conditioned with PA (P < 0.05). Treatment with PA resulted in more sealed interfaces than specimens treated with NRC. Within the limitations of this study the results showed that carbide burs leave a surface that is more conducive to bonding than diamond burs.

Decrease in inflammatory hyperalgesia by herpes vector-mediated knockdown of Nav1.7 sodium channels in primary afferents.

Induction of peripheral inflammation increases the expression of the Nav1.7 sodium channel in sensory neurons, potentially increasing their excitability. Peripheral inflammation also produces hyperalgesia in humans and an increase in nociceptive responsiveness in animals. To test the relationship between these two phenomena we applied a recombinant herpes simplex-based vector to the hindpaw skin of mice, which encoded both green fluorescent protein (GFP) as well as an antisense sequence to the Nav1.7 gene. The hindpaw was subsequently injected with complete Freund's adjuvant to induce robust inflammation. Application of the vector, but not a control vector encoding only GFP, prevented an increase in Nav1.7 expression in GFP-positive neurons and prevented development of hyperalgesia in both C and Adelta thermonociceptive tests. These results provide clear evidence of the involvement of an increased expression of the Nav1.7 channel in nociceptive neurons in the development of inflammatory hyperalgesia.

Peripheral and central p38 MAPK mediates capsaicin-induced hyperalgesia.

The stress-activated mitogen-activated protein kinase (MAPK) p38 is emerging as an important mediator of pain. The present study examined the possible involvement of peripheral and spinal p38 MAPK in capsaicin-induced thermal hyperalgesia. Topical capsaicin produced phosphorylation of p38 MAPK in the skin from the affected hindpaw as well as the corresponding lumbar spinal cord in a time dependent manner. Topical capsaicin produced robust C-fiber mediated thermal hyperalgesia that was inhibited by systemic, local peripheral, or central intrathecal pre-treatment with the p38 MAPK inhibitor, SD-282. Intraperitoneal SD-282 (10-60 mg/kg) significantly and dose-dependently attenuated capsaicin-induced C-fiber mediated thermal hyperalgesia. Similarly, 0.1-5mg/kg subcutaneous SD-282 in the hindpaw dose-dependently attenuated capsaicin-induced thermal hyperalgesia. Intrathecal administration of 1microg SD-282 was also anti-hyperalgesic in this model. Functionally, SD-282 decreased capsaicin-induced release of calcitonin gene related peptide in an in vitro skin release assay, consistent with a role for p38 MAPK in peripheral nerve function. These results suggest that p38 MAPK plays a role in the development of hyperalgesic states, exerting effects both centrally in the spinal cord and peripherally in sensory C fibers.

Aging affects two modes of nanoleakage expression in bonded dentin.

Water sorption into resin-dentin interfaces precedes hydrolytic degradation. We hypothesized that these processes are morphologically manifested by the uptake of ammoniacal silver nitrate, which is thought to trace hydrophilic domains and water-filled channels within matrices. Water sorption is thought to be nonuniform and can be traced by the use of silver nitrate. Human teeth bonded with an experimental filled-adhesive were aged in artificial saliva (experimental) or non-aqueous mineral oil (control). Specimens retrieved for up to a 12-month period were immersed in 50 wt% ammoniacal silver nitrate and examined by transmission electron microscopy for identification of the changes in their silver uptake. Reticular silver deposits initially identified within the bulk of hybrid layers in the experimental group were gradually reduced over time, but were subsequently replaced by similar deposits that were located along the hybrid layer-adhesive interface. Silver uptake in water-binding domains of the adhesive layers increased with aging, resulting in water tree formation. These water-filled channels may act as potential sites for hydrolytic degradation of resin-dentin bonds.

Polymeric system for dual growth factor delivery.

The development of tissues and organs is typically driven by the action of a number of growth factors. However, efforts to regenerate tissues (e.g., bone, blood vessels) typically rely on the delivery of single factors, and this may partially explain the limited clinical utility of many current approaches. One constraint on delivering appropriate combinations of factors is a lack of delivery vehicles that allow for a localized and controlled delivery of more than a single factor. We report a new polymeric system that allows for the tissue-specific delivery of two or more growth factors, with controlled dose and rate of delivery. The utility of this system was investigated in the context of therapeutic angiogenesis. We now demonstrate that dual delivery of vascular endothelial growth factor (VEGF)-165 and platelet-derived growth factor (PDGF)-BB, each with distinct kinetics, from a single, structural polymer scaffold results in the rapid formation of a mature vascular network. This is the first report of a vehicle capable of delivery of multiple angiogenic factors with distinct kinetics, and these results clearly indicate the importance of multiple growth factor action in tissue regeneration and engineering.

Quasisteady aero-acoustic response of orifices.

The low frequency response of orifices (slit, circular diaphragm, and perforated plate) in the presence of mean flow is well predicted by a quasisteady theory. A refinement is brought to the theory by considering a Mach number dependent vena contracta coefficient. The measurements of the vena contracta coefficient of a slit agree well with the simple analytical expression existing in the case of the Borda tube orifice. The measured scattering matrix coefficients do not depend strongly on the geometry of the element. If the frequency is increased the moduli remain relatively unaffected while the arguments exhibit a complex behavior which depends on the geometry. From these considerations an anechoic termination efficient at high mass flow is designed.

Minimally invasive operative care. II. Contemporary techniques and materials: an overview.

SUMMARY: Restorative dentistry has experienced a shift from the mainly reparative dentistry of the 20th century towards a minimal intervention approach. Contemporary operative treatment incorporates the MI philosophy in cavity design. Currently available techniques to pursue minimally invasive restorative treatments are highlighted. Characteristics of adhesive materials that facilitate minimally invasive operative care are discussed. CONCLUSION: When operative intervention is the designated treatment for initial caries, currently available operative techniques and contemporary materials warrant a minimally invasive approach. Minimal intervention applied to the operative field keeps the options open for long-term preservation of the restored tooth.

Minimally invasive operative care. I. Minimal intervention and concepts for minimally invasive cavity preparations.

SUMMARY: From the mainly reparative dentistry of the 20th century, contemporary dentistry shifts towards a minimal intervention (MI) approach encompassing up-to-date caries diagnosis and risk assessment before arriving at a treatment decision. An overview is provided of incorporating MI philosophy into the field of operative dentistry. The ultimate goal of MI is to extend the lifetime of restored teeth with as little intervention as possible. When operative care is indicated, it should be aimed at "prevention of extension." Black's principles for cavity design are considered and put in the perspective of minimally invasive operative care. Guiding principles for contemporary adhesive cavities are reviewed. CONCLUSION: Contemporary operative care should be based on a minimally invasive approach. Minimal intervention is not just a technique, it is a philosophy!

Engineering and characterization of functional human microvessels in immunodeficient mice.

SUMMARY: Current model systems used to investigate angiogenesis in vivo rely on the interpretation of results obtained with nonhuman endothelial cells. Recent advances in tissue engineering and molecular biology suggest the possibility of engineering human microvessels in vivo. Here we show that human dermal microvascular endothelial cells (HDMEC) transplanted into severe combined immunodeficient (SCID) mice on biodegradable polymer matrices differentiate into functional human microvessels that anastomose with the mouse vasculature. HDMEC were stably transduced with Flag epitope or alkaline phosphatase to confirm the human origin of the microvessels. Endothelial cells appeared dispersed throughout the sponge 1 day after transplantation, became organized into empty tubular structures by Day 5, and differentiated into functional microvessels within 7 to 10 days. Human microvessels in SCID mice expressed the physiological markers of angiogenesis: CD31, CD34, vascular cellular adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule 1 (ICAM-1). Human endothelial cells became invested by perivascular smooth muscle alpha-actin-expressing mouse cells 21 days after implantation. This model was used previously to demonstrate that overexpression of the antiapoptotic protein Bcl-2 in HDMEC enhances neovascularization, and that apoptotic disruption of tumor microvessels is associated with apoptosis of surrounding tumor cells. The proposed SCID mouse model of human angiogenesis is ideally suited for the study of the physiology of microvessel development, pathologic neovascular responses such as tumor angiogenesis, and for the development and investigation of strategies designed to enhance the neovascularization of engineered human tissues and organs.

Sustained release of vascular endothelial growth factor from mineralized poly(lactide-co-glycolide) scaffolds for tissue engineering.

Strategies to engineer bone tissue have focused on either: (1) the use of scaffolds for osteogenic cell transplantation or as conductive substrates for guided bone regeneration; or (2) release of inductive bioactive factors from these scaffold materials. This study describes an approach to add an inductive component to an osteoconductive scaffold for bone tissue engineering. We report the release of bioactive vascular endothelial growth factor (VEGF) from a mineralized, porous, degradable polymer scaffold. Three dimensional, porous scaffolds of the copolymer 85 : 15 poly(lactide-co-glycolide) were fabricated by including the growth factor into a gas foaming/particulate leaching process. The scaffold was then mineralized via incubation in a simulated body fluid. Growth of a bone-like mineral film on the inner pore surfaces of the porous scaffold is confirmed by mass increase measurements and quantification of phosphate content within scaffolds. Release of 125I-labeled VEGF was tracked over a 15 day period to determine release kinetics from the mineralized scaffolds. Sustained release from the mineralized scaffolds was achieved, and growth of the mineral film had only a minor effect on the release kinetics from the scaffolds. The VEGF released from the mineralized and non-mineralized scaffolds was over 70% active for up to 12 days following mineralization treatment, and the growth of mineral had little effect on total scaffold porosity.

Bioabsorbable polymer scaffolds for tissue engineering capable of sustained growth factor delivery.

Engineering new tissues utilizing cell transplantation on biodegradable polymer matrices is an attractive approach to treat patients suffering from the loss or dysfunction of a number of tissues and organs. The matrices must maintain structural integrity during the process of tissue formation, and promote the vascularization of the developing tissue. A number of molecules (angiogenic factors) have been identified that promote the formation of new vascular beds from endothelial cells present within tissues, and the localized, controlled delivery of these factors from a matrix may allow an enhanced vascularization of engineered tissues. We have developed a gas foaming polymer processing approach that allows the fabrication of three-dimensional porous matrices from bioabsorbable materials (e.g., copolymers of lactide and glycolide [PLG]) without the use of organic solvents or high temperatures. The effects of several processing parameters (e.g., gas type, polymer composition and molecular weight) on the process were studied. Several gases (CO(2), N(2), He) were utilized in the fabrication process, but only CO(2) resulted in the formation of highly porous, structurally intact matrices. Crystalline polymers (polylactide and polyglycolide) did not form porous matrices, while amorphous copolymers (50:50, 75:25, and 85:15 ratio of lactide:glycolide) foamed to yield matrices with porosity up to 95%. The mechanical properties of matrices were also regulated by the choice of PLG composition and molecular weight. Angiogenic factors (e.g., vascular endothelial growth factor) were subsequently incorporated into matrices during the fabrication process, and released in a controlled manner. Importantly, the released growth factor retains over 90% of its bioactivity. In summary, a promising system for the incorporation and delivery of angiogenic factors from three-dimensional, biodegradable polymer matrices has been developed, and the fabrication process allows incorporation under mild conditions.

Controlled growth factor release from synthetic extracellular matrices.

Polymeric matrices can be used to grow new tissues and organs, and the delivery of growth factors from these matrices is one method to regenerate tissues. A problem with engineering tissues that exist in a mechanically dynamic environment, such as bone, muscle and blood vessels, is that most drug delivery systems have been designed to operate under static conditions. We thought that polymeric matrices, which release growth factors in response to mechanical signals, might provide a new approach to guide tissue formation in mechanically stressed environments. Critical design features for this type of system include the ability to undergo repeated deformation, and a reversible binding of the protein growth factors to polymeric matrices to allow for responses to repeated stimuli. Here we report a model delivery system that can respond to mechanical signalling and upregulate the release of a growth factor to promote blood vessel formation. This approach may find a number of applications, including regeneration and engineering of new tissues and more general drug-delivery applications.