Digital cross-mounting of intraoral scan casts from a virtual articulator to a mechanical articulator by using a custom transfer plate: A dental technique.

With the development of digital dental technologies, a complete digital workflow without using physical casts has become possible. However, for certain clinical and dental laboratory procedures, especially in complex rehabilitation treatments, physically mounted casts in an ideal location in a mechanical articulator are still necessary for treatment planning and restoration fabrication. This technique report describes a digital approach to fabricating a custom transfer plate to cross mount intraoral scan casts from a virtual articulator to the corresponding mechanical articulator. This technique eliminates the need for conventional physical facebow transfer processes and offers a straightforward approach to integrating virtual procedures with analog workflows.

Superimposing digital arch scans onto cone beam computed tomography scans with metallic artifacts by applying a radiopaque occlusal registration material: A chairside dental technique.

Precise alignment between digital arch scans and cone beam computed tomography (CBCT) scans is a crucial step in computer-aided implant planning and placement. However, clinicians frequently encounter challenges during this process when imaging patients with existing metal restorations or orthodontic devices, as these can introduce metallic artifacts on CBCT scans that lead to alignment deviations. The presented technique describes a straightforward approach using a radiopaque occlusal registration material as a radiographic marker to facilitate the alignment between digital arch scans and CBCT scans with metallic artifacts. This technique simplifies the clinical workflow by eliminating the need for additional radiographic templates or specialized devices, offering a cost-effective option for clinicians.

A digital approach to fabricating a custom holder for the attachment of a mandibular sensor of an optical jaw motion tracking system: A dental technique.

A digital approach to fabricating a custom holder to attach a mandibular sensor of an optical jaw motion tracking system is described. Typically, jaw motion tracking systems come with standard holders. However, additional chairside time is still required to adapt the holder's arm to the individual arch and securely attach the holder to the mandibular teeth. Moreover, the placement of the standard holder is problematic in patients with a deep vertical overlap or with short clinical crowns. This technique offers a digital approach to designing and fabricating a custom holder in situations where standard holders cannot be efficiently attached. The custom holder is designed to accommodate the available space without interfering with the occlusion, thereby minimizing the time needed to attach the holder and optimizing the workflow for clinical jaw motion tracking.

A cast-free approach to fabricating an implant-supported interim restoration: A dental technique.

A cast-free approach is described to fabricate an implant-supported interim restoration by using the postoperative cone beam computed tomography (CBCT) scan to locate the placed implant. A postoperative intraoral scan was aligned to the postoperative CBCT scan through a dental implant planning software program. An attached interim abutment and implant analog complex was then scanned and superimposed on the placed implant in the postoperative CBCT scan. Once the best alignment was achieved, a virtual cast was generated, and an interim restoration with bilateral positioning wings was fabricated on the interim abutment and inserted during the second-stage surgery. This technique offers a cast-free approach to inserting an implant-supported interim restoration immediately after the second-stage surgery to guide the healing of the soft tissue that can minimize chairside time and optimize the clinical workflow.

Bone regeneration property of tooth-derived bone substitute prepared chairside for periodontal bone defects: an experimental study.

BACKGROUND: Periodontitis often leads to progressive destruction and loss of alveolar bone, the reconstruction of which remains difficult in periodontal therapy. As a novel bone graft material, tooth-derived bone substitute (TDBS) processed from extracted teeth has been previously reported about its osteoconductivity and promising results in bone regeneration. This study was to investigate the biological effects and bone regeneration properties of TDBS in vitro and in vivo using rat periodontal bone defect model. METHODS: Three groups of materials were used in the experiments: TDBS, TDBS treated with ethylene diamine tetraacetic acid (EDTA) (TDBS-E), and allogeneic bone materials. Calcium (Ca) and phosphate (P) ion dissolutions were quantified by spectrophotometer for seven days. The releases of bone morphogenetic protein-2 (BMP-2) and transforming growth factor-ß1 (TGF-ß1) were identified by enzyme-linked immunosorbent assay (ELISA). Human osteoblast proliferation, migration, and differentiation were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell counting, alkaline phosphatase activity (ALP), and alizarin red staining (ARS), respectively. Furthermore, the osteogenic effects of TDBS on periodontal furcation bone defects were evaluated at eight weeks postoperatively using micro-computed tomography (Micro-CT) and histological analysis. RESULTS: The dissolution of both Ca and P ions in TDBS increased over time. The BMP-2 released from TDBS was significantly higher than that from TDBS-E and allografts, while the TGF-ß1 release from TDBS and TDBS-E groups was higher than that in the allografts. The TDBS-E group could induce the highest level of osteoblast proliferation compared to other groups. Cell migration with allografts co-culture was significantly induced compared to the blank control. However, all groups demonstrated similar positive effects on osteoblast differentiation. Furthermore, in the periodontal model, all materials could effectively enhance bone regeneration in the furcation defect. CONCLUSIONS: The TDBS prepared chairside as an autogenous bone graft, demonstrating osteoinductivity, which enhances the osteogenic biological characteristics. Therefore, TDBS is suggested as an economical and biocompatible material for periodontal bone regeneration.

Cone-beam computed tomographic analysis of maxillary sinus septa among Yemeni population: a cross-sectional study.

BACKGROUND: Maxillary sinus septa increase perforation risk of Schneiderian membrane during the sinus floor elevation (SFE). Cone Beam Computed Tomography (CBCT) allows for a more precise assessment of the septal position; thus, preoperative CBCT analysis is substantial to avoid possible complications. This study aims to investigate the 3D characteristics of the maxillary sinus septa based on CBCT images. To our knowledge, no study reported the CBCT-based investigation for the sinus septa among Yemeni population. MATERIALS AND METHODS: This is a retrospective cross-sectional analysis of 880 sinus CBCT images 440 patients. The septa prevalence, locations, orientations, morphology, and associated factors were analyzed. The effect of age, gender, and dental status on the sinus septa and the relationship between sinus membrane pathology and sinus septa were also analyzed. Anatomage (Invivo version 6) was used for CBCT images analysis. Descriptive and analytical statistics were performed, and a P-value < 0.05 was significantly considered. RESULTS: The maxillary sinus septa were found among 63.9% of patients and 47% of sinuses. The average septa height was 5.2 mm. 15.7% of patients had septa in the right maxilla, 18% in the left, and 30.2% in both. Gender, age, and dental condition had no influence on the presence of septa, and septa presence did not influence sinus membrane pathology. Many septa originated from the floor (54.5%), located in the middle (43%), with coronal orientation (66%) and complete configuration (58.2%). CONCLUSION: Based on our findings, the septa prevalence, locations, orientations, and morphology were significant and equivalent to the highest recorded in the literature yet. Thus, when sinus floor elevation is planned, CBCT imaging of the maxillary sinus is recommended for safe dental implantation.

A digital approach to create a virtual implant preliminary cast for the fabrication of a splinted framework and custom open tray for multiple implants impression making: A dental technique.

A digital approach is described for fabricating a splinted framework and custom open tray for the impression of multiple implants based on a virtually created implant cast with the digital prediction of the future impression coping positions. In this technique, impression copings to be used in the definitive impression-making process were digitally scanned and aligned to a preliminary cast using installed protective caps of screw-retained abutments as references, which resulted in a virtual cast that offers information both about the impression coping positions and their surrounding tissues and could allow a splinted framework and custom open tray to be digitally designed and fabricated. Creation of a virtual cast through digital technologies eliminates the traditional manual splinted framework and custom tray fabrication procedures. Additionally, only one set of impression copings and analogs is needed in this technique, thereby reducing treatment time and cost. It also offers a splinted framework and custom open tray with consistent quality.

[Application of Animal Model Training in Preclinical Skills Teaching for Graduate Students of Dentistry].

Objective: To explore the potential application value of animal model training in improving the comprehensive clinical ability of postgraduate students of dentistry and to provide reference for new methods of preclinical skills teaching. Methods: A total of 40 postgraduate students of dentistry were assigned to two groups, an experimental group and a control group. The control group took the routine teaching course on root canal treatment for the right mandibular first molar, using a simulated model of human head. The experimental group also took a teaching course on root canal therapy for the right mandibular first molar, but an animal model was used for the group. After the course was completed, the instructor conducted comprehensive evaluation of the students' psychological quality, patient communication skills, diagnosis and treatment logic, speed of performing procedures, and treatment plan design. A questionnaire survey was conducted to examine the students' attitudes toward and evaluation of animal model training. Results: The scores for psychological quality (0.430±0.024 vs. 0.115±0.036), patient communication skills (0.878±0.065 vs. 0.115±0.036), diagnosis and treatment logic (0.630±0.066 vs. 0.372±0.033), speed of performing procedures (0.8975±0.019 vs. 0.055±0.080), and treatment plan design (0.539±0.036 vs. 0.396±0.017) of the experimental group were significantly higher than those of the control group ( P<0.0001). The total score of the experimental group (3.374±0.184) was significantly higher than that of the control group (1.053±0.082) and the difference was statistically significant ( P<0.001). 95% of the students in the control group and 100% of those in the experimental group were willing to participate in animal model training to improve their level of diagnosis and treatment skills for dental and endodontic diseases, showing no statistically significant difference ( χ 2=1.026, P=0.3112). In the experimental group, 30% of the students believed that their psychological qualities had been improved, 50% believed that their procedure skills had been improved, and 20% believed that animal model training had expanded the scope of their theoretical knowledge. Conclusion: Adding animal model training can improve dentistry graduate students' comprehensive abilities, including their psychological quality, patient communication skills, diagnosis and treatment logic, speed of performing procedures, and treatment plan design. In addition, it helps students familiarize themselves in advance with animal experimental operations for basic research, thus helping them acquire dual professional skills.

Investigation of membrane fouling and mechanism induced by extracellular organic matter during long-term exposure to pharmaceuticals and personal care products.

This study investigated ultrafiltration membrane fouling by extracellular organic matter (EOM) and the mechanism operating during long-term exposure to pharmaceuticals and personal care products. The results indicated that carbamazepine and diclofenac in algal-laden water altered the filtration flux and membrane fouling by EOM. Exposure to low-concentration carbamazepine (0.25 µg/L) improved the filtration flux and the total (Rtot) and reversible fouling resistance (Rc), whereas the filtration flux and Rtot and Rc were reduced when EOM was used during long-term exposure to high carbamazepine concentrations (>1 µg/L). Both Rtot and Rc were increased when algae were exposed to 0.25 µg/L diclofenac, whereas the filtration flux and Rtot and Rc were alleviated when algae were exposed to >1 µg/L diclofenac. Moreover, carbamazepine and diclofenac (0.25 µg/L - 1000 µg/L) in water enhanced the irreversible fouling resistance (Rb) when ultrafiltration was used to treat algal-laden waters. The mechanism indicated that membrane fouling induced by standard blocking was transformed to complete blocking when EOM was exposed to high levels of carbamazepine (>0.25 µg/L) in the initial filtration process, whereas cake layer formation played an important role during the later filtration process; with low carbamazepine levels (0.25 µg/L), standard blocking of EOM was dominant during the entire filtration process. The membrane fouling mechanism also changed when algal-laden waters were exposed to diclofenac, the membrane fouling was transformed from complete blocking to standard blocking when DFC was present in the initial filtration process, whereas cake layer formation exerted an important role during the late filtration process. This research provides important information on the long-term risks caused by pharmaceutical and personal care products and potential threats to membrane treatment.

An Indirect Digital Technique to Transfer 3D Printed Casts to a Mechanical Articulator With Individual Sagittal Condylar Inclination Settings Using CBCT and Intraoral Scans.

With the widespread application of digital impression techniques in prosthetic dentistry, accurate intraoral scan mounting, and virtual articulator parameters setting as per patients' anatomic structures are essential for treatment planning and restoration fabrication, especially for complex rehabilitation cases; meanwhile, marginal fit checking, occlusal adjustment, and porcelain layering of restorations are also crucial procedures in all cases in which the analog procedure to mount maxillary arches on a mechanical articulator is still required. This technique article presents an indirect digital approach that can first achieve virtual intraoral scan mounting and sagittal condylar inclination value setting of an Artex virtual articulator based on bony structures provided by a single cone beam computed tomography scan. It then facilitates the transfer of virtually mounted intraoral scans from the virtual articulator to the matched Artex mechanical articulator by relating a digitally scanned mounting plate of the Artex mechanical articulator to the virtual articulator, printing the intraoral scan and mounting plate scan assembly, and then mounting the printed casts on the mechanical articulator based on the printed mounting plate. This technique eliminates the conventional facebow transfer and protrusive bite registration procedures and offers a straightforward approach for the seamless integration of virtual environments and analog workflows into clinical practice. It aids in the design of restorations that are harmonious with the mandibular movements and reduces chairside adjustment time.

A Novel Technique to Align the Intraoral Scans to the Virtual Articulator and Set the Patient-Specific Sagittal Condylar Inclination.

Customized cast orientations and parameter settings of the virtual articulator according to the patient's condyles are indispensable parts of today's digital workflows in prosthodontics. This article describes a digital technique to align the intraoral scans to a virtual articulator by using a facial scanner to locate the patient's cutaneous landmarks of the arbitrary hinge axis and the reference plane, and to customize the sagittal condylar inclination of the virtual articulator through a digital protrusive interocclusal record and a dental computer-aided design software program. It enables individual cast orientations and virtual articulator parameter settings without conventional facebow transferring and bite registration procedures and can be easily integrated with most virtual articulator systems on the market to allow clinicians and technicians to work in a complete digital workflow and facilitate customized treatment planning and dental prosthesis fabrication.

[Effects of Three Illuminants on the Color of High Translucent Multilayered Zirconia].

OBJECTIVE: To evaluate the effects of three common illuminants on the color of four brands of high translucent multilayered zirconia (HTMZ) ceramics so as to provide reference for clinical practice and dental restoration fabrication, and to reduce the risks for illuminant change causing color mismatch between the natural teeth and the restorations made of HTMZ. METHODS: Four brands of commonly used HTMZ were selected and ten cuboid samples ( n=10/group) of 12 mm×10 mm×0.8 mm were prepared for each type of HTMZ. The L*, a*, and b* values of the samples were measured under D65, A and F2, three standard illuminants. Then, the L*, a*, and b* values were statistically analyzed by using factors of the type of the illuminant and the brand of the zirconia. Color difference (ΔE) of samples of the same brand under exposure to changed illuminants was also calculated. RESULTS: When the same samples were exposure to different illuminants, there was no significant difference in the L* value, the a* value for the different iluminants was shown to be illuminant A>illuminant F2>illuminant D65, and the b* value was shown to be illuminant F2>illuminant D65>illuminant A. The L*, a*, and b* values of samples of different brands showed statistically significant difference when they were exposed to the same illuminant ( P<0.0001). Samples of the same brand showed ΔE when they were under the three different illuninants, and all ΔE were clinically acceptable. CONCLUSION: The types of illuminant used, to a certain degree, affected the hue and chroma of HTMZ. There were colorimetric differences between restorations made of different brands of HTMZ ceramics of the same color. The types of illuminants most common to the daily life of patients and the color characteristics of materials of different brands should be taken into consideration to facilitate the selection of restoration materials and dental restoration fabrication, and to reduce the risks for color mismatch between the restorations and the adjacent teeth caused by the change of illuminants.

Revealing Ionic Signal Enhancement with Probe Grafting Density on the Outer Surface of Nanochannels.

Probe-modified nanopores/nanochannels are one of the most advanced sensors because the probes interact strongly with ions and targets in nanoconfinement and create a sensitive and selective ionic signal. Recently, ionic signals have been demonstrated to be sensitive to the probe-target interaction on the outer surface of nanopores/nanochannels, which can offer more open space for target recognition and signal conversion than nanoconfined cavities. To enhance the ionic signal, we investigated the effect of grafting density, a critical parameter of the sensing interface, of the probe on the outer surface of nanochannels on the change rate of the ionic signal before and after target recognition (ß). Electroneutral peptide nucleic acids and negatively charged DNA are selected as probes and targets, respectively. The experimental results showed that when adding the same number of targets, the ß value increased with the probe grafting density on the outer surface. A theoretical model with clearly defined physical properties of each probe and target has been established. Numerical simulations suggest that the decrease of the background current and the aggregation of targets at the mouth of nanochannels with increasing probe grafting density contribute to this enhancement. This work reveals the signal mechanism of probe-target recognition on the outer surface of nanochannels and suggests a general approach to the nanochannel/nanopore design leading to sensitivity improvement on the basis of relatively good selectivity.

Endogenous GDF11 regulates odontogenic differentiation of dental pulp stem cells.

Dental stem cell-based tooth regeneration is the futuristic treatment for missing teeth. Growth differentiation factor 11 (GDF11), a novel member of the TGF-beta superfamily, has been reported to play a critical role in regulating stem cell differentiation. However, the role of endogenous GDF11 during dental stem cell differentiation remains unknown. Here, we have shown that GDF11 was highly expressed in dental pulp tissues in both mouse and human. Knockdown of endogenous GDF11 in human dental pulp stem cells (hDPSCs) led to comparable proliferation and migration but attenuated odontogenic differentiation as evidenced by alkaline phosphatase and Alizarin Red S staining. In addition, transcriptional levels of odontogenic-related genes were significantly down-regulated according to real-time polymerase chain reaction. Mechanistically, we performed RNA sequencing analysis and found that silencing of endogenous GDF11 compromised the process of ossification and osteoblast differentiation, especially down-regulated transcription expression of Wnt pathway-specific genes. Immunofluorescence staining also showed diminished ß-catenin expression and nuclei accumulation after knockdown of endogenous GDF11 in hDPSCs. In summary, our results suggested that endogenous GDF11 positively regulate odontogenic differentiation of hDPSCs through canonical Wnt/ß-catenin signalling pathway.

Hyperglycemia-induced inflamm-aging accelerates gingival senescence via NLRC4 phosphorylation.

Inflamm-aging was recently affiliated with the progression of diabetic complications. Local cellular senescence together with senescence-associated secretory phenotype (SASP) are the main contributors to inflamm-aging. However, little is known about their involvement in diabetic periodontitis. Gingiva is the first line of host defense in the periodontium, and macrophages are key SASP-carrying cells. Here, we explored the molecular mechanism by which hyperglycemia drives the inflamm-aging in the gingival tissue of diabetic mice and macrophages. We demonstrated that hyperglycemia increased the infiltrated macrophage senescence in gingival tissue of diabetic mice. Simultaneously, hyperglycemia elevated the local burden of senescent cells in gingival tissue and induced the serum secretion of SASP factors in vivo Moreover, in vitro, high glucose induced macrophage senescence and SASP factors secretion through phosphorylation of NLRC4, which further stimulated the NF-κB/Caspase-1 cascade via an IRF8-dependent pathway. Deletion of NLRC4 or IRF8 abolished hyperglycemia-induced cellular senescence and SASP in macrophages. In addition, we found that treatment with metformin inhibited NLRC4 phosphorylation and remarkably decreased cellular senescence and SASP in the context of hyperglycemia. Our data demonstrated that hyperglycemia induces the development of inflamm-aging in gingival tissue and suggested that NLRC4 is a potential target for treatment of diabetes-associated complications.

In silico genome-wide identification of m6A-associated SNPs as potential functional variants for periodontitis.

Genetic variation is considered to affect the N6 -methyladenosine (m6A) RNA transcript modification, which is the most prevalent posttranscriptional messenger RNA modification. This study aimed to identify m6A-associated single-nucleotide polymorphisms (m6A-SNPs) that may affect m6A methylation from numerous periodontitis (PD) SNPs. We identified an abundance of m6A-SNPs by analyzing raw data of published PD genome-wide association studies and m6A-SNPs list from the m6AVar database. Other evidence was found in public databases for expression quantitative trait loci (eQTL) and differential gene expression analysis. Accordingly, 1938 m6A-SNPs were identified, 104 of which appeared to be associated with PD (p < .05) while 65 showed eQTL signals. Lastly, the leading SNP rs2723183 (p = 3.93E-07) was predicted to regulate local gene IL-37 expression in PD (p = 2.64E-05; in GSE10334) and change regulatory motif RXRA. In summary, dozens of PD-associated m6A-SNPs were identified and their possible functions were demonstrated in this study.

The role of USP34 in the fixation of titanium implants in murine models.

Ubiquitin-specific protease 34 (USP34), a member of the ubiquitin-specific protease family, regulates osteogenic differentiation of bone marrow mesenchymal stem cells via bone morphogenetic protein signaling. This study aimed to investigate the role of USP34 in fixation of titanium implants in mouse models. Eight-week-old Usp34-knockout (Prx1-Cre;Usp34f/f ) mice and their Usp34 wild-type (Usp34f/f ) control littermates were used. Experimental titanium implants were inserted into the distal ends of femurs and the edentulous area of maxillae. Two and four weeks after surgery, samples of femur and maxilla were obtained, and micro-computed tomography scanning, histomorphometric analyses, and push-in tests were performed on the samples. Compared with controls, Prx1-Cre;Usp34f/f mice showed reduced bone volume for both femurs and maxillae; a decreased femoral bone-implant contact ratio (BIC) at 2 wk [mean (standard error of the mean): 62.17% (2.15%) vs. 44.06% (3.45%)] and 4 wk [72.46% (1.61%) vs. 64.53% (1.93%)]; decreases in femoral bone volume fraction (BV/TV) and push-in resistance; and lower BIC and BV/TV of the maxillae. Taken together, our data demonstrate that specific deletion of Usp34 in mesenchymal stem cells impairs fixation of titanium implants in mice.

Salivary microbiome in patients undergoing hemodialysis and its associations with the duration of the dialysis.

BACKGROUND: Chronic kidney disease (CKD) patients, especially those with end stage renal disease (ESRD) undergoing hemodialysis (HD), exhibit high prevalence of periodontitis. This cross-sectional study aimed to investigate the periodontal status of HD patients and its relationship with salivary microbiome. METHODS: One hundred eight HD patients and one hundred healthy control individuals were recruited. They were subjected to periodontal examination followed by saliva samples collection for 16S rRNA gene sequencing. RESULTS: The HD patients were with worse periodontal health status, and exhibited higher salivary microbial diversity and lower richness. The periodontal pathogens were significantly enriched in the HD patients. The inferred functional analyze showed microbes enriched in the HD patients were mainly related to metabolism. Despite the periodontal status and overall structure of the microbiome were not significantly altered as the HD duration prolonged, the abundance of Lachnospiraceae [G-2] sp. |HMT_096| is positively correlated with the duration of HD and the community periodontal index (CPI). Five OTUs (operational taxonomic units) belonging to the phyla Firmicutes were enriched as the duration prolonged, and four OTUs originated from the phyla Proteobacteria were negatively related with the CPI index. ESRD patients undergoing HD exhibited microbiota structural, compositional and functional differences compared with the healthy controls. And the species changed as the duration of hemodialysis prolonged. CONCLUSIONS: End stage renal disease changes salivary microbiome and is a risk factor for oral dysbiosis.

Recent Progress in Time-Resolved Biosensing and Bioimaging Based on Lanthanide-Doped Nanoparticles.

Luminescent nanomaterials have attracted great attention in luminescence-based bioanalysis due to their abundant optical and tunable surface physicochemical properties. However, luminescent nanomaterials often suffer from serious autofluorescence and light scattering interference when applied to complex biological samples. Time-resolved luminescence methodology can efficiently eliminate autofluorescence and light scattering interference by collecting the luminescence signal of a long-lived probe after the background signals decays completely. Lanthanides have a unique [Xe]4fN electronic configuration and ladder-like energy states, which endow lanthanide-doped nanoparticles with many desirable optical properties, such as long luminescence lifetimes, large Stokes/anti-Stokes shifts, and sharp emission bands. Due to their long luminescence lifetimes, lanthanide-doped nanoparticles are widely used for high-sensitive biosensing and high-contrast bioimaging via time-resolved luminescence methodology. In this review, recent progress in the development of lanthanide-doped nanoparticles and their application in time-resolved biosensing and bioimaging are summarized. At the end of this review, the current challenges and perspectives of lanthanide-doped nanoparticles for time-resolved bioapplications are discussed.

Redefining Molecular Amphipathicity in Reversing the "Coffee-Ring Effect": Implications for Single Base Mutation Detection.

The "coffee ring effect" is a natural phenomenon wherein sessile drops leave ring-shaped structures on the solid surfaces upon drying. It drives a nonuniform deposition of suspended compounds on the substrates, which adversely affects many processes, including surface-assisted biosensing and molecular self-assembly. In this study, we describe how the coffee ring effect can be eliminated by controlling the amphipathicity of the suspended compounds, for example, DNA modified with hydrophobic dye. Specifically, nuclease digestion of the hydrophilic DNA end converts the dye-labeled molecule into an amphipathic molecule (one with comparably weighted hydrophobic and hydrophilic ends) and reverses the coffee ring effect and results in a uniform disk-shaped feature deposition of the dye. The amphipathic product decreases the surface tension of the sessile drops and induces the Marangoni flow, which drives the uniform distribution of the amphipathic dye-labeled product in the drops. As a proof of concept, this strategy was used in a novel enzymatic amplification method for biosensing to eliminate the coffee ring effect on a nitrocellulose membrane and increase assay reliability and sensitivity. Importantly, the reported strategy for eliminating the coffee ring effect can be extended to other sessile drop systems for potentially improving assay reliability and sensitivity.