Influence of the orthodontic bonding procedure on biofilm formation.

INTRODUCTION: In orthodontics, white spot lesions are a persistent and widespread problem caused by the demineralization of buccal tooth surfaces around bonded brackets. The remaining adhesive around the brackets leads to surface roughness, which might contribute to demineralization. The present in vitro study aimed to compare a conventional and a modern adhesive system (APC Flash-Free technology) for orthodontic brackets with regard to the adhesion of Streptococcus sobrinus, a leading caries pathogen. METHODS: This in vitro study included 20 premolar teeth and compared 10 APC Flash-Free adhesive-coated ceramic brackets (FF)with 10 conventionally bonded (CB) ceramic clarity brackets. Specimens were incubated in an S. sobrinus suspension for 3 h. To evaluate the bacterial formation, samples were analysed with a scanning electron microscope (SEM). Imaging software was used to quantify and statistically compare percentage values of colonization (PVC) in both groups' adhesion and transition areas. RESULTS: We found a significant difference in biofilm formation between the groups for the adhesive and transition areas. PVC in the adhesive area was approximately 10.3-fold greater for the CB group compared with the FF group (median: 3.2 vs 0.31; P < 0.0001). For the transition area, median PVC was approximately 2.4-fold greater for the CB group compared with the FF group (median: 53.17 vs 22.11; P < 0.01). CONCLUSIONS: There was a significantly lower level of S. sobrinus formation around the FF bracket system than there was surrounding the conventionally bonded group. This study suggests that the FF adhesive bracket system can help reduce the occurrence of bacterial growth around orthodontic brackets.

Photodynamic Suppression of Enterococcus Faecalis in Infected Root Canals with Indocyanine Green, TroloxTM and Near-Infrared Light.

Recently, our group showed that additional supplementation of Trolox™ (vitamin E analogue) can significantly enhance the antimicrobial photodynamic effect of the photosensitizer Indocyanine green (ICG). Up to now, the combined effect has not yet been investigated on Enterococcus faecalis in dental root canals. In the present in vitro study, eighty human root canals were inoculated with E. faecalis and subsequently subjected to antimicrobial Photodynamic Therapy (aPDT) using ICG (250, 500, 1000 µg/mL) and near-infrared laser light (NIR, 808 nm, 100 Jcm-2). Trolox™ at concentrations of 6 mM was additionally applied. As a positive control, irrigation with 3% NaOCl was used. After aPDT, root canals were manually enlarged and the collected dentin debris was subjected to microbial culture analysis. Bacterial invasion into the dentinal tubules was verified for a distance of 300 µm. aPDT caused significant suppression of E. faecalis up to a maximum of 2.9 log counts (ICG 250 µg/mL). Additional application of TroloxTM resulted in increased antibacterial activity for aPDT with ICG 500 µg/mL. The efficiency of aPDT was comparable to NaOCl-irrigation inside the dentinal tubules. In conclusion, ICG significantly suppressed E. faecalis. Additional application of TroloxTM showed only minor enhancement. Future studies should also address the effects of TroloxTM on other photodynamic systems.

Salivary Pellicle Formed on Dental Composites Evaluated by Mass Spectrometry-An In Situ Study.

(1) Background: In the oral environment, sound enamel and dental restorative materials are immediately covered by a pellicle layer, which enables bacteria to attach. For the development of new materials with repellent surface functions, information on the formation and maturation of salivary pellicles is crucial. Therefore, the present in situ study aimed to investigate the proteomic profile of salivary pellicles formed on different dental composites. (2) Methods: Light-cured composite and bovine enamel samples (controls) were exposed to the oral cavity for 30, 90, and 120 min. All samples were subjected to optical and mechanical profilometry, as well as SEM surface evaluation. Acquired pellicles and unstimulated whole saliva samples were analyzed by SELDI-TOF-MS. The significance was determined by the generalized estimation equation and the post-hoc bonferroni adjustment. (3) Results: SEM revealed the formation of homogeneous pellicles on all test and control surfaces. Profilometry showed that composite surfaces tend to be of higher roughness compared to enamel. SELDI-TOF-MS detected up to 102 different proteins in the saliva samples and up to 46 proteins in the pellicle. Significant differences among 14 pellicle proteins were found between the composite materials and the controls. (4) Conclusions: Pellicle formation was material- and time-dependent. Proteins differed among the composites and to the control.

Influence of Different Bracket Adhesive Systems on Enamel Demineralization-An In Vitro Study.

BACKGROUND: enamel demineralization is a common side effect of orthodontic therapy with fixed braces. The aim of the present in vitro study was to compare a conventional adhesive system and a modern adhesive system (APC Flash-Free [FF] technology) with regard to the demineralization of enamel by Streptococcus sobrinus (S. sobrinus). METHODS: this in vitro study included premolar teeth and compared APC FF adhesive brackets (Group A, n = 15) with conventional adhesive brackets (Group B, n = 15) from the same company. Specimens were incubated with a positive control group (PCG, n = 5) and a negative control group (NCG, n = 5) in an S. sobrinus suspension for three weeks. To evaluate the grade of enamel demineralization, the samples were analyzed using a polarizing microscope. RESULTS: the test specimens of group B with conventionally bonded bracket adhesive showed significantly greater (+10.8 µm) demineralization with regard to the penetration depth of the demineralization than the PCG (p = 0.012). Thus, there was a difference from group A with the new bracket adhesive of the FF brackets (+7.29 µm). Significantly, demineralization was more pronounced cervically than coronally in both groups, and it occurred cervically more frequently than grade 3 demineralization (p = 0.001). CONCLUSIONS: it seems plausible that new orthodontic bracket adhesives and the modern FF adhesive system positively contribute to the reduction in enamel demineralization during orthodontic treatment.

Histological and Histomorphometric Evaluation of Implanted Photodynamic Active Biomaterials for Periodontal Bone Regeneration in an Animal Study.

Recently, our group developed two different polymeric biomaterials with photodynamic antimicrobial surface activity for periodontal bone regeneration. The aim of the present study was to analyze the biocompatibility and osseointegration of these materials in vivo. Two biomaterials based on urethane dimethacrylate (BioM1) and tri-armed oligoester-urethane methacrylate (BioM2) that additionally contained ß-tricalcium phosphate and the photosensitizer mTHPC (meso-tetra(hydroxyphenyl)chlorin) were implanted in non-critical size bone defects in the femur (n = 16) and tibia (n = 8) of eight female domestic sheep. Bone specimens were harvested and histomorphometrically analyzed after 12 months. BioM1 degraded to a lower extent which resulted in a mean remnant square size of 17.4 mm², while 12.2 mm² was estimated for BioM2 (p = 0.007). For BioM1, a total percentage of new formed bone by 30.3% was found which was significant higher compared to BioM2 (8.4%, p < 0.001). Furthermore, BioM1 was afflicted by significant lower soft tissue formation (3.3%) as compared to BioM2 (29.5%). Additionally, a bone-to-biomaterial ratio of 81.9% was detected for BioM1, while 8.5% was recorded for BioM2. Implantation of BioM2 caused accumulation of inflammatory cells and led to fibrous encapsulation. BioM1 (photosensitizer-armed urethane dimethacrylate) showed favorable regenerative characteristics and can be recommended for further studies.

Coaxial electrospun nanofibers as drug delivery system for local treatment of periodontitis.

OBJECTIVES: The aim of the present study was to prepare resorbable polylactide fibers for periodontitis treatment using coaxial electrospinning to optimize the release of metronidazole (MNA) by reducing the initial burst effect. METHODS: Poly(L-lactide-co-D,L-lactide) (PLA) fibers mats with different distributions of metronidazole (MNA) were manufactured by coaxial electrospinning (COAX). By COAX spinning the central core of the fiber was enriched with 40% MNA (m/m), while the sheath of the fiber consisted of PLA only (test group). In contrast, fibers of the control group were prepared by conventional electrospinning with the same amount of MNA but with a homogenous drug distribution (HDD - homogenously distributed drug). The release of MNA was determined by analyzing aliquots from the fiber mats using UV-VIS spectroscopy. Agar diffusion tests were carried out to determine the antibacterial effect on periodontopathogenic bacteria. Biocompatibility was tested in direct contact to human gingival fibroblasts (HGF) for two days. RESULTS: The COAX mats showed a retarded drug release compared to the conventional HDD fibers. After 24 h, 64% of total MNA was released cumulatively from the COAX fibers while 90% of the MNA was released from the HDD fibers (controls). The antibacterial effect of COAX fibers was significantly higher after 24 h compared to the HDD fibers. Cell cultivation revealed significant higher numbers of vital cells among the COAX mats. SIGNIFICANCE: COAX fibers showed improved sustained MNA release compared to conventional fibers and can be seen as potential drug delivery systems in local periodontitis treatment.

Remineralization of Artificially Demineralized Human Enamel and Dentin Samples by Zinc-Carbonate Hydroxyapatite Nanocrystals.

(1) Background: Decalcified enamel and dentin surfaces can be regenerated with non-fluoride-containing biomimetic systems. This study aimed to investigate the effect of a zinc carbonate-hydroxyapatite-containing dentifrice on artificially demineralized enamel and dentin surfaces. (2) Methods: Human enamel and dentin discs were prepared and subjected to surface demineralization with 30% orthophosphoric acid for 60 s. Subsequently, in the test group (n = 20), the discs were treated three times a day for 3 min with a zinc carbonate-hydroxyapatite-containing toothpaste (biorepair®). Afterwards, all samples were gently rinsed with PBS (5 s) and stored in artificial saliva until next use. Samples from the control group (n = 20) received no dentifrice-treatment and were stored in artificial saliva, exclusively. After 15 days of daily treatment, specimens were subjected to Raman spectroscopy, energy-dispersive X-ray micro-analysis (EDX), white-light interferometry, and profilometry. (3) Results: Raman spectroscopy and white-light interferometry revealed no significant differences compared to the untreated controls. EDX analysis showed calcium phosphate and silicon dioxide precipitations on treated dentin samples. In addition, treated dentin surfaces showed significant reduced roughness values. (4) Conclusions: Treatment with biorepair® did not affect enamel surfaces as proposed. Minor mineral precipitation and a reduction in surface roughness were detected among dentin surfaces only.

Antibacterial Effect of Endodontic Disinfections on Enterococcus Faecalis in Dental Root Canals-An In-Vitro Model Study.

Enterococcus faecalis (E. faecalis) is rather unsusceptible to many root canal disinfections which often cause a therapeutic problem. Therefore, the present in vitro study observed the efficiency of different endodontic antiseptics in their capability to suppress E. faecalis, especially inside dentinal tubules. Prior to any testing, root canals of extracted third human molars were inoculated with E. faecalis for 48 h. Antiseptic dressings with chloramine-T or calcium hydroxide (CaOH) for 24 h or irrigations with 1.3% sodium hypochlorite (NaOCl) were applied with n = 10 in each group. As control irrigation with normal saline was used. All treated canals were manually enlarged from size ISO 50 to 110 and the ablated dentin debris was subjected to microbial culture analysis. Bacterial colonization of the dentinal tubules up to 300 µm was verified by scanning electron microscopy and histological sample preparation. Application of crystalline chloramine-T caused total bacterial suppression inside the dentinal tubules. Dressings with CaOH showed only minor effects. Irrigation with NaOCl caused total eradication of bacteria adhering to the root canal walls, but also failed to completely suppress E. faecalis inside the dentinal tubules. The study showed that chloramine-T is of strong antiseptic activity and also efficient in suppressing E. faecalis inside dentinal tubules.

Antimicrobial Effect of Natural Berry Juices on Common Oral Pathogenic Bacteria.

(1) Background: Antimicrobial agents such as chlorhexidine (CHX) are commonly used in oral plaque control. However, sometimes those agents lack antimicrobial efficiency or cause undesired side effects. To identify alternative anti-infective agents, the present study investigated the antibacterial activity of all-fruit juices derived from blackcurrant, redcurrant, cranberry and raspberry on common oral pathogenic gram-positive and gram-negative bacteria (Streptococcus mutans, Streptococcus gordonii, Streptococcus sobrinus, Actinomyces naeslundii, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Enterococcus faecalis). (2) Methods: Antibacterial efficiency was evaluated by agar diffusion assay and in direct contact with bacteria in planktonic culture. Furthermore, cytotoxicity on human gingival fibroblasts was determined. (3) Results: Blackcurrant juice was most efficient at suppressing bacteria; followed by the activity of redcurrant and cranberry juice. Raspberry juice only suppressed P. gingivalis significantly. Only high-concentrated blackcurrant juice showed minimal cytotoxic effects which were significantly less compared to the action of CHX. (4) Conclusion: Extracts from natural berry juices might be used for safe and efficient suppression of oral pathogenic bacterial species.

Bactericidal and Biocompatible Properties of Plasma Chemical Oxidized Titanium (TiOB®) with Antimicrobial Surface Functionalization.

Coating of plasma chemical oxidized titanium (TiOB®) with gentamicin-tannic acid (TiOB® gta) has proven to be efficient in preventing bacterial colonization of implants. However, in times of increasing antibiotic resistance, the development of alternative antimicrobial functionalization strategies is of major interest. Therefore, the aim of the present study is to evaluate the antibacterial and biocompatible properties of TiOB® functionalized with silver nanoparticles (TiOB® SiOx Ag) and ionic zinc (TiOB® Zn). Antibacterial efficiency was determined by agar diffusion and proliferation test on Staphylocuccus aureus. Cytocompatibility was analyzed by direct cultivation of MC3T3-E1 cells on top of the functionalized surfaces for 2 and 4 d. All functionalized surfaces showed significant bactericidal effects expressed by extended lag phases (TiOB® gta for 5 h, TiOB® SiOx Ag for 8 h, TiOB® Zn for 10 h). While TiOB® gta (positive control) and TiOB® Zn remained bactericidal for 48 h, TiOB® SiOx Ag was active for only 4 h. After direct cultivation for 4 d, viable MC3T3-E1 cells were found on all surfaces tested with the highest biocompatibility recorded for TiOB® SiOx Ag. The present study revealed that functionalization of TiOB® with ionic zinc shows bactericidal properties that are comparable to those of a gentamicin-containing coating.

Study of energy transfer by different light curing units into a class III restoration as a function of tilt angle and distance, using a MARC Patient Simulator (PS).

OBJECTIVE: The MARC Patient Simulator (MARC PS) enables researchers to observe the influence of handling errors on the radiant exposure that is delivered by light curing units (LCUs). Changes in the tilt angle and distance of the light guide exit face in relation to the surface of the composite increment have a distinct effect on the total amount of light delivered during polymerization and may cause insufficient conversion of the material. Therefore, the aim of the present study was to determine the influence of the tilt angle and distance of irradiance on the efficiency of light application by recording the total amount of energy using the anterior tooth sensor of the MARC PS. METHODS: The influence of the tilt angle and distance of the light guide to the sensor surface on the delivered radiant exposure was examined for three different LCUs (Celalux 2 [C2], Bluephase [BP] and Translux Powerblue [TPB]). The measurements were performed for 20 s each with five different tilt angles (α=0°, 5°, 10°, 15°, 20°) and nine different distances (L=1, 2, 3, 4, 5, 6, 7, 8, 9 mm). RESULTS: For all LCUs, a distinct influence of the tilt angle on the delivered amount of fluence was found. At 0° tilt the C2 delivered a total light energy of 38.55 J/cm(2). By increasing the tilt of the light guide the amount of energy applied significantly decreased. At 20° tilt a reduction by 31.2% of the original light fluence was recorded. However, the C2 was the most powerful LCU measured. Even under optimum measurement conditions, the BP delivered a fluence of only 14.8 J/cm(2). At a tilt angle of 20°, though, the light sensor still registered 92.7% of the original output power. Under the same conditions, the TPB delivered 81.4%. With increasing distance of the light guide exit face to the surface of the sensor all LCUs showed a significant loss in delivered light energy. At a distance of 2mm the C2 showed a reduction by 46.7%, whereas total fluence of BP and TPB were reduced by 3.8% and 4.8%, respectively. SIGNIFICANCE: The choice of LCU and the application of an appropriate curing time are important for successful polymerization. Nevertheless, a perpendicular positioning of the light guide as close as possible to the surface of the composite increment is of essential importance in order to ensure sufficient delivery of light.

Antibacterial photodynamic treatment of periodontopathogenic bacteria with indocyanine green and near-infrared laser light enhanced by Trolox(TM).

BACKGROUND AND OBJECTIVES: It has been shown that certain vitamins can significantly enhance the effect of photodynamic anti-tumor therapy. Unfortunately, there is no sufficient information available about the impact of those antioxidants on antimicrobial Photodynamic Therapy (aPDT). The present study is aimed at investigating the antimicrobial effect of the dye indocyanine green (ICG) in the presence of Trolox(TM) , a vitamin E analogue, upon irradiation with near-infrared (NIR) laser light (808 nm) on the gramnegative periodontopathogenic bacteria Aggregatibacter actinomycetemcomitans (A.a.), Porphyromonas gingivalis (P.g.) and Fusobacterium nucleatum (F.n.). METHODS: Bacteria solved in PBS were incubated with ICG (50-500 µg/ml) in the presence and absence of Trolox(TM) (2 mM). Irradiation was performed after 10 minutes of dark-incubation with NIR-laser-light (25-100 J/cm(2) , 810 nm). During treatment, temperature was also recorded inside the bacterial solutions. The treated suspensions were serial diluted and plated onto blood agar plates. After anaerobe cultivation for 5 days the colony-forming units (CFU/ml) were determined. RESULTS: The antibacterial effect was ICG-concentration and exposure dependent. It was found that high ICG-concentrations and light fluence rates caused bacterial reduction due to hyperthermia. Where low ICG-concentrations (<250 µg/ml) and fluence rates only induced minor regression, additional Trolox(TM) -administration significantly enhanced the photodynamic effect. While treatment of A.a. (250 µg/ml ICG, 100 J/cm(2) ) without Trolox(TM) caused no bacterial reduction, additional administration led to total eradication. In the presence of Trolox(TM) reduction to one-fifth of the original ICG-concentration (50 µg/ml) still induced total suppression of P.g. and F.n. at identical fluence (100 J/cm(2) ). Treatment with ICG, NIR-light or Trolox(TM) alone showed no remarkable bactericidal effect. Application of high ICG-concentrations (500 µg/ml) and exposure values (100 J/cm(2) ) caused peak temperatures of 64.53°C. CONCLUSIONS: The results clearly show that Trolox(TM) significantly enhanced the antibacterial effect of ICG upon irradiation with NIR-laser-light. Additional administration of Trolox(TM) may also increase the efficiency of other aPDT systems.

Colonization of Enterococcus faecalis in a new SiO/SiO(2)-microtube in vitro model system as a function of tubule diameter.

OBJECTIVES: Endodontic pathogens can penetrate deep into dentinal tubules and therefore survive the chemo-mechanical disinfection procedures. Bacterial penetration has been mainly studies using sliced infected human teeth which, besides creating artifacts, can hinder the observation of the inner tubules due to the dense and opaque dentin structure. The aim of the present study was to develop a standardized dentin model by using artificial SiO/SiO2-microtubes of different diameters and lengths to test the penetration ability of Enterococcus faecalis. METHODS: E. faecalis was grown in Schaedler fluid media for 24h and thereafter cell density was settled to 10(3)cells/ml by addition of fresh media. The bacterial solution was then incubated for 2, 3, 5 and 10 days with the SiO/SiO2-microtubes of different diameters (2-5.5µm) and lengths (100-500µm). The colonization of the tubes was evaluated by phase-contrast microscopy and the amount of colonization was determined by using a colonization index (CI; 0-none, 1-mild, 2-moderate, 3-heavy). RESULTS: The diameter of the tubes strongly influences the microbial colonization. After 2 days of cultivation the 5.5µm tubes showed a moderate to heavy colonization (CI 2-3). In comparison, the 2 and 3µm tubes were clearly less colonized at the same point in time. In detail: at day 3, only mild to moderate bacteria colonization (CI 1-2) were found in the 3µm tubes and at day 10 penetration of the 2µm tubes just started. The colonization of the 5.5µm tubes was also influenced by their length. In case of the longer microtubes, though, a smaller share of heavily colonized tubes was observed. SIGNIFICANCE: Our results show that E. faecalis was able to penetrate and reproduce within the standardized SiO/SiO2-microtubes in a short time. To examine the mechanisms of bacterial adhesion and invasion into tubular structures the 2µm tubes could serve as a model system because the diameters are similar to those of dentinal tubules.

Cytocompatibility of polymer-based periodontal bone substitutes in gingival fibroblast and MC3T3 osteoblast cell cultures.

OBJECTIVES: Inflammatory periodontal diseases are accompanied by destruction of periodontal tissue and alveolar bone. Infrabony lesions can be regenerated with adequate bone substitutes, which require high biocompatibility of the material. METHODS: To rate the biocompatibility of nine polymeric periodontal bone substitutes (Bio 1-Bio 9), cell viability and cytotoxicity assays were performed. For viability, human gingival fibroblasts (HGFs) and MC3T3 osteoblasts were cultured on the bone substitutes. For cytotoxicity, biomaterial extracts were prepared by incubation with culture medium for maximally 28days, and cells were exposed to the extracts for 1day. Polymers Bio 1 to Bio 5 were prepared by solvent casting, Bio 6 to Bio 9 by photopolymerization of the monomers at wavelengths of 400-500nm in the presence of a suitable photoinitiation system. RESULTS: Bio 1, Bio 3, Bio 4, Bio 5, and Bio 7 showed moderate to excellent cytocompatibility for both HGFs and osteoblasts in viability tests. Together with the results of the cytotoxicity assays, four of the nine tested polymers were considered cytocompatible: Bio 1 (poly(vinyl butyral-co-vinyl alcohol-co-vinyl acetate; PVB)), Bio 4 and Bio 5 (functionalized oligolactones), and, to a limited degree, Bio 7 (urethane methacrylate). Except for Bio 7, the cytocompatible polymers showed intermediate water contact angles (74-85°) and therefore moderate to low hydrophilicity. SIGNIFICANCE: The non-cross-linked polymers Bio 1, Bio 4, or Bio 5, and the photopolymerized polymeric network Bio 7 display good/excellent cytocompatibility and are therefore potential candidates for tissue engineering in alveolar bone substitution.

Photodynamic suppression of Enterococcus faecalis using the photosensitizer mTHPC.

BACKGROUND AND OBJECTIVES: Enterococcus faecalis is frequently found in persistent endodontic infections. In this context, the antimicrobial photodynamic therapy (aPDT) could become a modern alternative to existing antibacterial treatment approaches. The aim of this study was to investigate the effect of aPDT on E. faecalis using the photosensitizer (PS) 5,10,15,20-tetra(m-hydroxyphenyl)chlorin (mTHPC) enriched in liposomes. MATERIALS AND METHODS: Enterococcus faecalis was cultivated in Schaedler submerged culture for 24 hours, then isolated and adjusted in PBS to 10(8) cells/ml. The bacterial suspension was pipetted into a black microtitration plate and incubated for 15 minutes in the dark with mTHPC in various concentrations (10, 30, and 50 µM). The photosensitized suspensions were subjected to laser light (652 nm) at a light fluence of 100 J cm(-2) (test group A). In addition, the suspension sensitized with 50 µM mTHPC was irradiated with 25, 50, and 75 J cm(-2) (test group B). The following controls were used: non-irradiated bacterial suspension in the absence of mTHPC (C); irradiated bacterial suspension in the absence of mTHPC (D); non-irradiated bacterial suspension incubated with mTHPC (E). Dilution series (10(0)-10(-6)) were made of all groups and applied on Schaedler agar. After anerobic cultivation (4 days), the colony-forming units (CFU/ml) were determined. RESULTS: Enterococcus faecalis was suppressed completely after incubation with 50 µM mTHPC and illumination with 100 J cm(-2). Photodynamic treatment with 10 and 30 µM mTHPC caused reduction in CFU by 5.8 and 6.7 log-units. The application of an energy fluence <100 J cm(-2) resulted in a decline of antibacterial efficiency. Irradiation of the non-photosensitized solution showed no suppressing impact. Incubation of the PS without additional irradiation caused a maximal reduction in CFU by 1.5 log-units. CONCLUSION: The results show that aPDT using the PS mTHPC incorporated in liposomes could be a new approach to adjuvant treatment of endodontic infections with E. faecalis.

Structural alterations of spiny stellate cells in the somatosensory cortex in ephrin-A5-deficient mice.

Previous studies demonstrated that in ephrin-A5-deficient mice corticothalamic arbors are reduced by more than 50% in layer 4 of the somatosensory cortex (S1), where ephrin-A5 is normally expressed. Here we examined possible consequences of the reduced thalamic input on spiny stellate cells, the target neurons of thalamocortical afferents. Using ballistic delivery of particles coated with lipophilic dyes in fixed slices and confocal laser-microscopy, we could quantitatively analyze the morphology of these neurons. Cells were examined in S1 at postnatal day 8 (P8), when thalamic afferents establish synaptic contacts and the dendrites of their target cells are covered with filopodia, and at P23, after synapse formation and replacement of filopodia by spines. Our results indicate that at P8 the dendrites of cells in mutant animals exhibit more filopodia and are more branched than dendrites of wildtype cells. In contrast, there is no difference in the extent of the dendritic tree between knockout and control animals. At P23, dendrites of neurons in ephrin-A5-deficient mice are still more branched, but possess fewer spines than wildtype cells. Thus, at early stages layer 4 neurons appear to compensate the reduced thalamic input by increasing dendritic branching and the density of filopodia. However, while at later stages the dendrites of layer 4 neurons in mutants are still more branched, their spine density is now lower than in wildtype cells. Taken together, these data demonstrate that the structure of spiny stellate cells is shaped by thalamic input and Eph receptor signaling.