A Randomized Clinical Trial to Assess Caries Arrest by Using Silver Diamine Fluoride in U.S. Children: Interim Findings.

Purpose: To assess the effectiveness of 38 percent silver diamine fluoride (SDF) in arresting cavitated caries lesions in young U.S. children. Methods: Children 12 to 71 months of age with severe early childhood caries participated in this phase three, multicenter, randomized, placebocontrolled trial. SDF was applied twice (at baseline and six months), and children were followed for eight months. A planned interim analysis of only the six-month primary outcome caries arrest data, for approximately half of the cohort (680 of 1,144 children), was conducted using a generalized estimating equation model, accounting for non-independence among carious lesions within a patient. Results: Five hundred ninety-nine of the 680 participants, with 1,413 lesions, completed the six-month exam. Lesions in the SDF group demonstrated 54 percent arrest versus 21 percent in the placebo (P<0.001). Conclusions: Silver diamine fluoride was effective at arresting active cavitated lesions in this population, leading to the early stop of the trial. Final analyses of all data and other outcomes are currently underway.

Oral Health in the Down Syndrome Population: Parental Perceptions on Dental Care in the United States.

Purpose: To assess the oral health status of children with Down syndrome (DS) in the United States and evaluate the oral health needs of families with DS. Methods: Among 320 parents who consented to participate, 260 (81.2 percent) surveys were completed. A survey was distributed through the National Down Syndrome Society to parents of children with DS up to age 21 years, which asked questions about children's general and oral health. Results: Parents who reported that their children had difficulty rinsing and spitting were more likely to report their child's overall general health as poor (P<0.05). Parents' frequency of daily toothbrushing was similar to their children's toothbrushing habits (P<0.05). Conclusions: Dysphagia for children with Down syndrome may negatively impact oral health in addition to general health. Encouraging parental involvement in oral care for children with DS may lower their risk for oral disease. Continued support is needed to ensure dental school education includes training for the treatment and management of patients with DS.

Investigating the effects of arginine methylation inhibitors on microdissected brain tumour biopsies maintained in a miniaturised perfusion system.

Arginine methylation is a post-translational modification that consists of the transfer of one or two methyl (CH3) groups to arginine residues in proteins. Several types of arginine methylation occur, namely monomethylation, symmetric dimethylation and asymmetric dimethylation, which are catalysed by different protein arginine methyltransferases (PRMTs). Inhibitors of PRMTs have recently entered clinical trials to target several types of cancer, including gliomas (NCT04089449). People with glioblastoma (GBM), the most aggressive form of brain tumour, are among those with the poorest quality of life and likelihood of survival of anyone diagnosed with cancer. There is currently a lack of (pre)clinical research on the possible application of PRMT inhibitors to target brain tumours. Here, we set out to investigate the effects of clinically-relevant PRMT inhibitors on GBM biopsies. We present a new, low-cost, easy to fabricate perfusion device that can maintain GBM tissue in a viable condition for at least eight days post-surgical resection. The miniaturised perfusion device enables the treatment of GBM tissue with PRMT inhibitors ex vivo, and we observed a two-fold increase in apoptosis in treated samples compared to parallel control experiments. Mechanistically, we show thousands of differentially expressed genes after treatment, and changes in the type of arginine methylation of the RNA binding protein FUS that are consistent with hundreds of differential gene splicing events. This is the first time that cross-talk between different types of arginine methylation has been observed in clinical samples after treatment with PRMT inhibitors.

The Challenges and Rewards of Teledentistry.

On March 15, 2020, routine dental care in New York State paused due to the COVID-19 pandemic. The pause lasted 10 weeks in part to preserve critical supplies of personal protective equipment (PPE). This interruption of access to dental care led to an overall deterioration of oral health, an increase in prescribing and use of antibiotics and analgesic medications, especially opiates, and a rise in visits to hospital emergency centers. New York University's College of Dentistry, an academic ambulatory dental center, normally sees over 1,000 patient visits per day. Most visits are patients who require urgent care or are in the process of treatment to restore debilitating oral health problems. NYU Dentistry responded to the State pause by creating a nascent teledentistry service that began operations on March 17, 2020.

A systematic review and meta-analysis on chloroquine and hydroxychloroquine as monotherapy or combined with azithromycin in COVID-19 treatment.

Many recent studies have investigated the role of either Chloroquine (CQ) or Hydroxychloroquine (HCQ) alone or in combination with azithromycin (AZM) in the management of the emerging coronavirus. This systematic review and meta-analysis of either published or preprint observational studies or randomized control trials (RCT) aimed to assess mortality rate, duration of hospital stay, need for mechanical ventilation (MV), virologic cure rate (VQR), time to a negative viral polymerase chain reaction (PCR), radiological progression, experiencing drug side effects, and clinical worsening. A search of the online database through June 2020 was performed and examined the reference lists of pertinent articles for in-vivo studies only. Pooled relative risks (RRs), standard mean differences of 95% confidence intervals (CIs) were calculated with the random-effects model. Mortality was not different between the standard care (SC) and HCQ groups (RR = 0.99, 95% CI 0.61-1.59, I2 = 82%), meta-regression analysis proved that mortality was significantly different across the studies from different countries. However, mortality among the HCQ + AZM was significantly higher than among the SC (RR = 1.8, 95% CI 1.19-2.27, I2 = 70%). The duration of hospital stay in days was shorter in the SC in comparison with the HCQ group (standard mean difference = 0.57, 95% CI 0.20-0.94, I2 = 92%), or the HCQ + AZM (standard mean difference = 0.77, 95% CI 0.46-1.08, I2 = 81). Overall VQR, and that at days 4, 10, and 14 among patients exposed to HCQ did not differ significantly from the SC [(RR = 0.92, 95% CI 0.69-1.23, I2 = 67%), (RR = 1.11, 95% CI 0.26-4.69, I2 = 85%), (RR = 1.21, 95% CI 0.70-2.01, I2 = 95%), and (RR = 0.98, 95% CI 0.76-1.27, I2 = 85% )] respectively. Exposure to HCQ + AZM did not improve the VQR as well (RR = 3.23, 95% CI 0.70-14.97, I2 = 58%). The need for MV was not significantly different between the SC and HCQ (RR = 1.5, 95% CI 0.78-2.89, I2 = 81%), or HCQ + AZM (RR = 1.27, 95% CI 0.7-2.13, I2 = 88%). Side effects were more reported in the HCQ group than in the SC (RR = 3.14, 95% CI 1.58-6.24, I2 = 0). Radiological improvement and clinical worsening were not statistically different between HCQ and SC [(RR = 1.11, 95% CI 0.74-1.65, I2 = 45%) and (RR = 1.28, 95% CI 0.33-4.99), I2 = 54%] respectively. Despite the scarcity of published data of good quality, the effectiveness and safety of either HCQ alone or in combination with AZM in treating COVID-19 cannot be assured. Future high-quality RCTs need to be carried out.PROSPERO registration: CRD42020192084.

Differential regulation of Ca2+ influx by ORAI channels mediates enamel mineralization.

Store-operated Ca2+ entry (SOCE) channels are highly selective Ca2+ channels activated by the endoplasmic reticulum (ER) sensors STIM1 and STIM2. Their direct interaction with the pore-forming plasma membrane ORAI proteins (ORAI1, ORAI2, and ORAI3) leads to sustained Ca2+ fluxes that are critical for many cellular functions. Mutations in the human ORAI1 gene result in immunodeficiency, anhidrotic ectodermal dysplasia, and enamel defects. In our investigation of the role of ORAI proteins in enamel, we identified enamel defects in a patient with an ORAI1 null mutation. Targeted deletion of the Orai1 gene in mice showed enamel defects and reduced SOCE in isolated enamel cells. However, Orai2-/- mice showed normal enamel despite having increased SOCE in the enamel cells. Knockdown experiments in the enamel cell line LS8 suggested that ORAI2 and ORAI3 modulated ORAI1 function, with ORAI1 and ORAI2 being the main contributors to SOCE. ORAI1-deficient LS8 cells showed altered mitochondrial respiration with increased oxygen consumption rate and ATP, which was associated with altered redox status and enhanced ER Ca2+ uptake, likely due to S-glutathionylation of SERCA pumps. Our findings demonstrate an important role of ORAI1 in Ca2+ influx in enamel cells and establish a link between SOCE, mitochondrial function, and redox homeostasis.

The Impact of an Interprofessional Pediatric Oral Health Clerkship on Advancing Interprofessional Education Outcomes.

The aim of this study was to evaluate the effectiveness of an innovative pediatric interprofessional education clinical experience using oral-systemic health as the clinical population example for improving the self-reported interprofessional competencies of family nurse practitioner, dental, and medical students. The objectives of the interprofessional experience were for students to apply pediatric oral health assessment, identify the pediatric oral-systemic connection, and practice a team-based approach to improve oral-systemic outcomes. In spring 2015, fall 2015, and spring 2016, a total of 162 family nurse practitioner, dental, and medical students participated in this interprofessional experience at Bellevue Pediatric Outpatient Clinics together with a pediatric dental resident. Team members collaborated in reviewing the patient chart, taking the patient's medical and dental history, performing an oral assessment, applying fluoride varnish, and providing education and anticipatory guidance. The Interprofessional Collaborative Competency Attainment Survey (ICCAS) was used as a pretest and posttest to evaluate the degree to which students perceived changes in their attitudes about interprofessional competencies following the learning experience. In the results, all students had improved mean scores from pretest to posttest after the experience, and these changes were statistically significant for all students: nurse practitioner (p<0.01), dentistry (p<0.01), and medicine (p<0.001). The mean change from pretest to posttest was statistically significant for each of the six interprofessional competency domains (p<0.01). In both pediatric dental and primary care settings, the changes from pre- to posttest were significant (p<0.001). The experience was similarly effective for all groups of students in increasing their attitudes about interprofessional collaboration. These findings suggest that a clinical approach can be an effective strategy for helping health professions students develop interprofessional competence.

Rescue of Premature Coronal Suture Fusion with TGF-ß2 Neutralizing Antibody in Rabbits with Delayed-Onset Synostosis.

OBJECTIVES: An overexpression of Tgf-ß2 leads to calvarial hyperostosis and suture fusion in individuals with craniosynostosis. Inhibition of Tgf-ß2 may help rescue fusing sutures and restore normal growth. The present study was designed to test this hypothesis. DESIGN: Twenty-eight New Zealand White rabbits with delayed-onset coronal synostosis had radiopaque markers placed on either side of the coronal sutures at 10 days of age. The rabbits were randomly assigned to: (1) sham control rabbits (n = 10), (2) rabbits with control IgG (100 µg/suture) delivered in a collagen vehicle (n = 9), and (3) rabbits with Tgf-ß2 neutralizing antibody (100 µg/suture) delivered in a collagen vehicle (n = 9). Longitudinal growth data were collected at 10, 25, 42, and 84 days of age. Sutures were harvested at 84 days of age for histomorphometry. RESULTS: Radiographic analysis showed significantly greater ( P < .05) coronal suture marker separation, craniofacial length, cranial vault length, height, shape indices, cranial base length, and more lordotic cranial base angles in rabbits treated with anti-Tgf-ß2 antibody than in controls at 42 and 84 days of age. Histologically, rabbits treated with anti-Tgf-ß2 antibody at 84 days of age had patent and significantly ( P < .05) wider coronal sutures and greater sutural area compared to controls. CONCLUSIONS: These data support our hypothesis that antagonism of Tgf-ß2 may rescue fusing coronal sutures and facilitate craniofacial growth in this rabbit model. These findings also suggest that cytokine therapy may have clinical significance in infants with progressive postgestational craniosynostosis.

Efficient in vivo gene delivery using modified Tat peptide with cationic lipids.

A combination of modified HIV-1 Tat (mTat) peptide and cationic lipids, FuGENE HD (FH), dramatically enhanced transfection efficiency across a range of cell lines when compared to mTat or FH alone (Biomaterials 35:1705-1715 2014). The efficiency of this Tat peptide combination was significantly higher than many commercial non-viral vectors. In this present study, we tested the feasibility of this non-viral vector, mTat/FH, in vivo using plasmid DNA encoding a luciferase gene. The results of the in vivo studies showed that animals administered mTat/FH/DNA intramuscularly had significantly higher and longer luciferase expression (≈7 months) than those with mTat/DNA, FH/DNA, or DNA alone. Histological evaluation showed little immune response in the muscles, livers, and kidneys of mice administered with the mTat/FH. The combination of mTat with FH could significantly improve transfection efficiency, expanding the potential use of non-viral gene vectors in vivo.

The effect of a bioactive collagen membrane releasing PDGF or GDF-5 on bone regeneration.

Regenerative procedures using barrier membrane technology are presently well established in periodontal/endodontic surgery. The objective of this study was to compare the subsequent effects of the released platelet-derived growth factor (PDGF) and growth/differentiation factor 5 (GDF-5) from collagen membranes (CMs) on bone regeneration in vitro and in vivo. In vitro studies were conducted using MC3T3-E1 mouse preosteoblasts cultured with or without factors. Cell viability, cell proliferation, alkaline phosphatase (ALP) activity and bone marker gene expression were then measured. In vivo studies were conducted by placing CMs with low or high dose PDGF or GDF-5 in rat mandibular defects. At 4 weeks after surgery new bone formation was measured using µCT and histological analysis. The results of in vitro studies showed that CM/GDF-5 significantly increased ALP and cell proliferation activities without cytotoxicity in MC3T3-E1 cells when compared to CM/PDGF or CM alone. Gene expression analysis revealed that Runx2 and Osteocalcin were significantly increased in CM/GDF-5 compared to CM/PDGF or control. Quantitative and qualitative µCT and histological analysis for new bone formation revealed that although CM/PDGF significantly enhanced bone regeneration compared to CM alone or control, CM/GDF-5 significantly accelerated bone regeneration to an even greater extent than CM/PDGF. The results also showed that GDF-5 induced new bone formation in a dose-dependent manner. These results suggest that this strategy, using a CM carrying GDF-5, might lead to an improvement in the current clinical treatment of bone defects for periodontal and implant therapy.

Long-term efficient gene delivery using polyethylenimine with modified Tat peptide.

Polyethylenimine (PEI), a cationic polymer, has been widely studied and shown great promise as an efficient gene delivery vehicle. Likewise, the HIV-1 Tat peptide, a cell-permeable peptide, has been successfully used for intracellular gene delivery. To improve the favorable properties of these two vectors, we combine PEI with the modified Tat peptide sequence bearing histidine and cysteine residues (mTat). In vitro mTat/PEI-mediated transfection was evaluated by luciferase expression plasmid in two cell types. mTat/PEI produced significant improvement (≈5-fold) in transfection efficiency of both cell lines with little cytotoxicity when compared to mTat alone, PEI alone, or four commercial reagents. The particle size of mTat/PEI/DNA complex was significantly smaller than mTat or PEI alone, and it was correlated with higher transfection efficiency. Filipin III, an inhibitor of caveolae-mediated endocytosis, significantly inhibited mTat/PEI transfection. In contrast, chlorpromazine, an inhibitor of clathrin-mediated endocytosis, did not. This suggested caveolae-mediated endocytosis as the transfection mechanism. Furthermore, the results of in vivo studies showed that animals administered mTat/PEI/DNA intramuscularly had significantly higher and longer luciferase expression (≈7 months) than those with mTat/DNA, PEI/DNA, or DNA alone, without any associated toxicity. The combination of mTat with PEI could significantly improve transfection efficiency, expanding the potential use as a non-viral gene vector both in vitro and in vivo.

Delivery of Transforming Growth Factor-ß3 Plasmid in a Collagen Gel Inhibits Cranial Suture Fusion in Rats.

Objective : Studies described in this paper were designed to test the hypothesis that an increase in nonviral, plasmid-encoded Tgf-ß3 production, localized to the rat posterior frontal suture, prevents programmed suture fusion. Design : We developed a gene delivery system based on a dense collagen gel to deliver nonviral plasmids that encode for Tgf-ß3. Studies were performed to test the ability of this system to rescue rat cranial suture fusion in vitro and in vivo. Immunohistochemical studies were conducted to characterize the possible mechanisms by which increased production and presence of Tgf-ß3 protein interferes with suture fusion. Results : Posterior frontal sutures in the Tgf-ß3 plasmid-treated group exhibited 77% to 85% less bony bridging than the collagen control and untreated groups after 15 days in culture. In animals treated with Tgf-ß3 plasmid or Tgf-ß3 protein, there was a significant reduction in suture fusion in the middle region of the posterior frontal sutures when compared with control groups. In this region the Tgf-ß3 plasmid-treated group revealed 70% to 75% less bony bridging than control groups in vivo. Conclusions : Collagen gel can be formulated to provide release of nonviral plasmid DNA that results in cell transfection and elevated Tgf-ß3 protein production. Tgf-ß3 is an important regulator of suture fusion, and an increase in plasmid-encoded Tgf-ß3 protein is effective in inhibiting programmed suture fusion in rats.

Identification and long term stability of DNA captured on a dental impression wafer.

PURPOSE: The purpose of this study was to determine the quantity and quality of DNA extracted from a dental bite impression wafer immediately after impression and after 12 months of home storage. The authors' hypothesis was that the wafer would retain sufficient DNA with appropriate genetic markers to make an identification match. METHODS: Two impression wafers (Toothprints(®) brand) were administered to 100 3- to 26-year-olds. A cotton swab was used as a control. DNA from wafers stored for 12 months at home were compared to DNA collected at time 0 and compared to swabs at specific sites to determine quality and accuracy. The amount of DNA captured and recovered was analyzed using MagAttract technology and a quantitative real-time polymerase chain reaction. Capillary gel electrophoresis was performed to determine the quality of the DNA profiles obtained from the wafers vs those generated from the swabs of each subject. RESULTS: Average DNA concentration was: 480 pg/µL (wafer at time 0); 392 pg/µL (wafer after 12 months kept by subjects); and 1,041 pg/µL (buccal swab). Sufficient DNA for human identification was recovered from all sets of wafers, producing clear DNA profiles and accurate matches to buccal swabs. No inhibitors were found that could interfere with DNA profiling. CONCLUSIONS: Toothprints® impression wafers can be useful for DNA collection and child identification. After 12 months, the wafer was still usable for DNA capture and identification match.

Relaxin does not rescue coronal suture fusion in craniosynostotic rabbits.

OBJECTIVES: Craniosynostosis affects 1 in 2000 to 3000 live births and may result in craniofacial and neural growth disturbances. Histological data have shown that thick collagenous bundles are present in the sutural ligament, which may tether the osteogenic fronts, resulting in premature fusion. The hormone relaxin has been shown to disrupt collagen fiber organization, possibly preventing craniosynostosis by relaxing the sutural ligament and allowing osteogenic fronts to separate normally and stay patent. This study tested this hypothesis with a rabbit model of delayed-onset coronal suture synostosis. METHODS: A total of 18 New Zealand White rabbits with craniosynostosis were randomly assigned to one of three groups: sham control, protein control (BSA), relaxin treatment. After initial diagnosis, sham surgery, BSA, or relaxin was delivered to the fusing coronal suture in a slow-release (56-day) collagen vehicle. Longitudinal radiographs and body weights were collected at 10, 25, 42, and 84 days of age, and sutures were harvested for histology. RESULTS: Relaxin-treated animals had more disorganized intrasuture content than control groups. These specimens also appeared to have relatively wider sutures ectocranially. There were no significant differences in relaxin-treated animals for all craniofacial growth measures, or suture separation compared with controls. CONCLUSIONS: These data do not support our initial hypothesis that the use of relaxin may rescue sutures destined to undergo premature suture fusion. These findings suggest that collagen fiber arrangement may not be important for suture fusion. This protein therapy would not be clinically useful for craniosynostosis.

Blocking bone morphogenetic protein function using in vivo noggin therapy does not rescue premature suture fusion in rabbits with delayed-onset craniosynostosis.

BACKGROUND: Craniosynostosis is defined as the premature fusion of one or more cranial sutures. Bone morphogenetic proteins (BMPs), regulators of ossification, have been implicated in premature suture fusion. Noggin, an extracellular BMP inhibitor, has been shown experimentally to inhibit resynostosis following surgery. The present study was designed to test the hypothesis that BMP inhibition using noggin therapy may rescue sutures destined to fuse by inhibiting initial ossification. METHODS: Twenty-six, 10-day old rabbits with familial, delayed-onset, coronal suture synostosis were randomly divided into three groups: (1) the sham surgical control group, (2) the bovine serum albumin-treated group [10 µg/suture (protein/vehicle controls)], and (3) the noggin therapy group (10 µg/suture; experimental group). Sutural growth was monitored by radiopaque markers implanted at 10 days of age. At 25 days, the bovine serum albumin or noggin was combined with a slow-resorbing collagen vehicle and injected subperiosteally above the coronal suture. Somatic and sutural growth data were collected at 10, 25, 42, and 84 days of age. Coronal sutures were harvested at 84 days to histologically assess fusion. RESULTS: Results showed no significant (p > 0.05) differences in suture separation at any age. Suture fusion assessed by histomorphology did not differ among the three groups. Although previous data showed noggin to inhibit postoperative resynostosis in this craniosynostotic rabbit model, here there was no effect on initial suture fusion. CONCLUSION: These results suggest that in this rabbit model of craniosynostosis, BMPs do not play a role in the pathogenesis of craniosynostosis and only play a role in postoperative bony wound healing.

Modified Tat peptide with cationic lipids enhances gene transfection efficiency via temperature-dependent and caveolae-mediated endocytosis.

The HIV-1 Tat peptide has been successfully used for intracellular gene delivery. Likewise, various lipid-based methods have shown increased endocytosis and can influence endosomal escape. This study combines the favorable properties of Tat peptide with that of lipid systems for DNA delivery. We combined the lipid FuGENE HD (FH) with the Tat peptide sequence modified with histidine and cysteine residues (mTat). mTat/FH transfection was evaluated by luciferase expression plasmid in five cell types. mTat/FH produced significant improvement in transfection efficiency of all cell lines when compared to FH or mTat. Treatment with chloroquine, associated with energy-dependent endocytosis, significantly increased transfection efficiency with mTat/FH while incubation at low temperature decreased it. The zeta potential of mTat/FH/DNA was significantly higher compared to FH, mTat, or their DNA combination in the presence of serum, and it was correlated with transfection efficiency. The particle size of the FH/DNA complex was significantly reduced by addition of mTat. Filipin III, an inhibitor of caveolae-mediated endocytosis, significantly inhibited mTat/FH transfection, but transfection was increased by chlorpromazine, an inhibitor of clathrin-mediated endocytosis. These findings demonstrated the feasibility of using a combination of mTat with lipids, utilizing temperature-dependent and caveolae-mediated endocytosis, as a potentially attractive non-viral gene vector.

Use of mobile electronic devices as educational tool in pediatric community outreach.

The introduction of mobile electronic devices, as opposed to paper forms, in pediatric outreach programs of the New York University College of Dentistry is discussed. Since 2007, students have been receiving training on how to operate a personal digital assistant (PDA) and use it in community outreach for non-invasive oral-facial screenings and patient education. The shift from using paper forms to electronic media had a positive impact among the academic community, as it resulted in saving time and reducing the possibility of data collection errors. It may represent a significant improvement in data collection and patient education; and it provides an opportunity to enhance research and quality assessment.

Nutrition and oral health considerations in children with special health care needs: implications for oral health care providers.

Children with special health care needs are at increased risk for oral diseases. The purpose of this article was to discuss: nutritional and oral health factors routinely observed in most chronic childhood disorders; dietary modifications associated with select systemic disorders and how they may impact oral health in children; and the following factors common to chronic disorders associated with diet modifications-decreased appetite and increased nutritional risk; frequency of food intake; parental overindulgence; long-term use of cariogenic medications; and xerostomia. Characteristics of childhood disorders that require dietary modifications (congenital heart disease, cystic fibrosis, cancer, AIDS/HIV, diabetes mellitus, and phenylketonuria) are summarized. In addition, healthy dietary modifications and oral health recommendations are suggested. Implementation of these recommendations can assist the dentist and dental team as they join physicians and nutritionists in delivering the best possible care to children with special health care needs.

Comparison of transfection efficiency of nonviral gene transfer reagents.

This study compared six commercially available reagents (Arrest-In, ExpressFect, FuGENE HD, jetPEI, Lipofectamine 2000, and SuperFect) for gene transfection. We examined the efficiency and cytotoxicity using nine different cell lines (MC3T3-E1 mouse preosteoblasts, PT-30 human epithelial precancer cells, C3H10T1/2 mouse stem cells, MCF-7 human breast cancer cells, HeLa human cervical cancer, C2C12 mouse myoblasts, Hep G2 human hepatocellular carcinoma, 4T1 mouse mammary carcinoma, and HCT116 human colorectal carcinoma), and primary cells (HEKn human epidermal keratinocytes) with two different plasmid DNAs encoding luciferase or ß-galactosidase in the presence or absence of serum. Maximal transfection efficiency in MC3T3-E1, C3H10T1/2, HeLa, C2C12, Hep G2, and HCT116 was seen using FuGENE HD, in PT-30, 4T1, and HEKn was seen using Arrest-In, and in MCF-7 was seen using jetPEI. Determination of cytotoxicity showed that the largest amount of viable cells was found after transfection with jetPEI and ExpressFect. These results suggest that FuGENE HD is the most preferred transfection reagent for many cell lines, followed by Arrest-In and jetPEI. These results may be useful for improving nonviral gene and cell therapy applications.

Comparison of craniofacial phenotype in craniosynostotic rabbits treated with anti-Tgf-beta2 at suturectomy site.

OBJECTIVE: Overexpression of transforming growth factor-beta 2 has been associated with craniosynostosis and resynostosis following surgery. We examined the effects of localized transforming growth factor-beta 2 inhibition on craniofacial phenotype in rabbits with craniosynostosis. DESIGN: Twenty-five New Zealand white rabbits with bilateral coronal craniosynostosis were divided into three treatment groups: (1) suturectomy control (n=8); (2) suturectomy with nonspecific, control immunoglobulin G antibody (n=6); and (3) suturectomy with anti-transforming growth factor-beta 2 antibody (n=11). At 10 days of age, a coronal suturectomy was performed on all rabbits. The sites in groups 2 and 3 were immediately filled with a slow-resorbing collagen gel mixed with either immunoglobulin G or anti-transforming growth factor-beta 2 antibody. Computed tomography scans of each rabbit were acquired at ages 10, 25, and 84 days. Craniofacial landmarks were collected from three-dimensional computed tomography reconstructions, and growth and form were compared among the three groups. RESULTS: Rabbits treated with anti-transforming growth factor-beta 2 antibody differed in form at 84 days of age compared with suturectomy control rabbits, specifically in the snout and posterior neurocranium. Growth in some areas of the skull was greater in rabbits from the anti-transforming growth factor-beta 2 group than in suturectomy control rabbits, but not significantly greater than in IgG control rabbits. CONCLUSIONS: We find support for the hypothesis that transforming growth factor-beta 2 inhibition alters adult form, but these changes do not appear to be localized to the suturectomy region. Slight differences in form and growth between the two control groups suggest that the presence of the collagen vehicle itself may affect skull growth.