Anti-cancer effect of bee venom on human MDA-MB-231 breast cancer cells using Raman spectroscopy.

We demonstrated the apoptotic effect of bee venom (BV) on human MDA-MB-231 breast cancer cells using Raman spectroscopy and principal component analysis (PCA). Biochemical changes in cancer cells were monitored following BV treatment; the results for different concentrations and treatment durations differed markedly. Significantly decreased Raman vibrations for DNA and proteins were observed for cells treated with 3.0 µg/mL BV for 48 h compared with those of control cells. These results suggest denaturation and degradation of proteins and DNA fragmentation (all cell death-related processes). The Raman spectroscopy results agreed with those of atomic force microscopy and conventional biological tests such as viability, TUNEL, and western blot assays. Therefore, Raman spectroscopy, with PCA, provides a noninvasive, label-free tool for assessment of cellular changes on the anti-cancer effect of BV.

Observations of the Effects of Angiotensin II Receptor Blocker on Angiotensin II-Induced Morphological and Mechanical Changes in Renal Tubular Epithelial Cells Using Atomic Force Microscopy.

OBJECTIVE: Angiotensin II (Ang II) plays a profibrotic role in the kidneys. Although many pathways of Ang II have been discovered, the morphological and mechanical aspects have not been well investigated. We observed the changes in tubular epithelial cells (TECs) after Ang II treatment with or without Ang II receptor blockers (ARBs) using atomic force microscopy (AFM). METHODS: TECs were stimulated with Ang II with or without telmisartan, PD123319, and blebbistatin. AFM was performed to measure the cellular stiffness, cell volume, and cell surface roughness. Epithelial to mesenchymal transition markers were determined via immunocytochemistry. RESULTS: After Ang II stimulation, cells transformed to a flattened and elongated mesenchymal morphology. Cell surface roughness and volume significantly increased in Ang II treated TECs. Ang II also induced an increase in phospho-myosin light chain and F-actin and a decrease in E-cadherin. Ang II coincubation with either telmisartan or blebbistatin attenuated these Ang II-induced changes. CONCLUSION: We report, for the first time, the use of AFM in directly observing the changes in TECs after Ang II treatment with or without ARBs. Simultaneously, we successfully measured the selective effect of PD123319 or blebbistatin. AFM could be a noninvasive evaluating strategy for cellular processes in TECs.

Effects of polymer-coated boron nitrides with increased hemorheological compatibility on human erythrocytes and blood coagulation.

BACKGROUND: Boron nitride (BN) nanomaterials are promising in biomedical research owing to their large surface area, graphene-like structure, and chemical and thermal properties. However, the toxicological effects of BN on erythrocytes and blood coagulation remain uninvestigated. OBJECTIVE: The aims of our study were to synthesize glycol chitosan (GC)- and hyaluronic acid (HA)-coated BNs, and to investigate the effects of these BNs on human cancer cells, erythrocytes, and whole blood. METHODS: We prepared hemocompatible forms of BN coated with GC and HA, and evaluated them using cell uptake/viability tests, hemolysis analysis and FE-SEM, as well as through hemorheological evaluation methods such as RBC deformability and aggregation, and blood coagulation. RESULTS: GC/BN and HA/BN were both ∼200 nm, were successfully taken into cells, and emitted blue fluorescence. Both BNs were less toxic than bare BN, even at higher concentrations. The aggregation index of human red blood cells (RBCs) after 2 h incubation with BN, GC/BN, and HA/BN was greatly influenced, whereas RBC deformability did not dramatically change. CONCLUSIONS: We found that GC/BN affected the intrinsic coagulation pathway, whereas both GC/BN and HA/BN affected the extrinsic pathway. Therefore, HA/BN is less detrimental to RBCs and blood coagulation dynamics than bare BN and GC/BN.

Physicochemical and surface properties of acrylic intraocular lenses and their clinical significance.

To analyze and compare several commercially available acrylic intraocular lenses (IOLs) with particular regard to their clinical significance, we examined the physicochemical and surface properties of four currently available acrylic IOLs using static water contact angle, atomic force microscopy (AFM), Raman spectroscopy, and differential scanning calorimetry (DSC) measurements. The hydrophobic acrylic IOLs, ZA9003, and MA60BM, had contact angles ranging from 77.9° ± 0.65° to 84.4° ± 0.09°. The contact angles in the hydrophilic acrylic (970C) and heparin-surface-modified (HSM) hydrophilic acrylic IOLs (BioVue) were 61.8° ± 0.45° and 69.7° ± 0.76°, respectively. The roughness of the IOL optic surface differed depending on the type of IOL (p < 0.001). The surface roughness of BioVue had the lowest value: 5.87 ± 1.26 nm. This suggests that the BioVue IOL may lead to reduced cellular adhesion compared to the unmodified IOLs. All IOLs including those composed of acrylic optic materials from different manufacturers showed distinct Raman spectra peaks. The glass transition temperatures (Tg) for the hydrophobic acrylic IOLs were between 12.5 and 13.8 °C. These results suggest that the intraoperative and postoperative behavior of an IOL can be predicted. This information is also expected to contribute greatly to the industrial production of reliable biocompatible IOLs.

Label-free Detection for a DNA Methylation Assay Using Raman Spectroscopy.

BACKGROUND: DNA methylation has been suggested as a biomarker for early cancer detection and treatment. Varieties of technologies for detecting DNA methylation have been developed, but they are not sufficiently sensitive for use in diagnostic devices. The aim of this study was to determine the suitability of Raman spectroscopy for label-free detection of methylated DNA. METHODS: The methylated promoter regions of cancer-related genes cadherin 1 (CDH1) and retinoic acid receptor beta (RARB) served as target DNA sequences. Based on bisulfite conversion, oligonucleotides of methylated or nonmethylated probes and targets were synthesized for the DNA methylation assay. Principal component analysis with linear discriminant analysis (PCA-DA) was used to discriminate the hybridization between probes and targets (methylated probe and methylated target or nonmethylated probe and nonmethylated target) of CDH1 and RARB from nonhybridization between the probe and targets (methylated probe and nonmethylated target or nonmethylated probe and methylated target). RESULTS: This study revealed that the CDH1 and RARB oligo sets and their hybridization data could be classified using PCA-DA. The classification results for CDH1 methylated probe + CDH1 methylated target versus CDH1 methylated probe + CDH1 unmethylated target showed sensitivity, specificity, and error rates of 92%, 100%, and 8%, respectively. The classification results for the RARB methylated probe + RARB methylated target versus RARB methylated probe + RARB unmethylated target showed sensitivity, specificity, and error rates of 92%, 93%, and 11%, respectively. CONCLUSIONS: Label-free detection of DNA methylation could be achieved using Raman spectroscopy with discriminant analysis.

Evaluation of non-thermal plasma-induced anticancer effects on human colon cancer cells.

Non-thermal atmospheric-pressure plasma has been introduced in various applications such as sterilization, wound healing, blood coagulation, and other biomedical applications. The most attractive application of non-thermal atmospheric-pressure plasma is in cancer treatment, where the plasma is used to produce reactive oxygen species (ROS) to facilitate cell apoptosis. We investigate the effects of different durations of exposure to dielectric-barrier discharge (DBD) plasma on colon cancer cells using measurement of cell viability and ROS levels, western blot, immunocytochemistry, and Raman spectroscopy. Our results suggest that different kinds of plasma-treated cells can be differentiated from control cells using the Raman data.

The Role of Free Radicals in Hemolytic Toxicity Induced by Atmospheric-Pressure Plasma Jet.

Atmospheric-pressure plasma (APP) has received attention due to its generation of various kinds of reactive oxygen/nitrogen species (ROS/RNS). The controllability, as well as the complexity, is one of the strong points of APP in various applications. For biological applications of this novel method, the cytotoxicity should be estimated at various levels. Herein, we suggest red blood cell (RBC) as a good cell model that is simpler than nucleated cells but much more complex than other lipid model systems. Air and N2 gases were compared to verify the main ROS/RNS in cytotoxicity, and microscopic and spectroscopic analyses were performed to estimate the damages induced on RBCs. The results shown here will provide basic information on APP-induced cytotoxicity at cellular and molecular levels.

Low-Cost Label-Free Biosensing Bimetallic Cellulose Strip with SILAR-Synthesized Silver Core-Gold Shell Nanoparticle Structures.

We introduce a label-free biosensing cellulose strip sensor with surface-enhanced Raman spectroscopy (SERS)-encoded bimetallic core@shell nanoparticles. Bimetallic nanoparticles consisting of a synthesis of core Ag nanoparticles (AgNP) and a synthesis of shell gold nanoparticles (AuNPs) were fabricated on a cellulose substrate by two-stage successive ionic layer absorption and reaction (SILAR) techniques. The bimetallic nanoparticle-enhanced localized surface plasmon resonance (LSPR) effects were theoretically verified by computational calculations with finite element models of optimized bimetallic nanoparticles interacting with an incident laser source. Well-dispersed raspberry-like bimetallic nanoparticles with highly polycrystalline structure were confirmed through X-ray and electron analyses despite ionic reaction synthesis. The stability against silver oxidation and high sensitivity with superior SERS enhancement factor (EF) of the low-cost SERS-encoded cellulose strip, which achieved 3.98 × 108 SERS-EF, 6.1%-RSD reproducibility, and <10%-degraded sustainability, implicated the possibility of practical applications in high analytical screening methods, such as single-molecule detection. The remarkable sensitivity and selectivity of this bimetallic biosensing strip in determining aquatic toxicities for prohibited drugs, such as aniline, sodium azide, and malachite green, as well as monitoring the breast cancer progression for urine, confirmed its potential as a low-cost label-free point-of-care test chip for the early diagnosis of human diseases.

Applications of Microfluidic Devices for Urology.

Microfluidics is considered an important technology that is suitable for numerous biomedical applications, including cancer diagnosis, metastasis, drug delivery, and tissue engineering. Although microfluidics is still considered to be a new approach in urological research, several pioneering studies have been reported in recent years. In this paper, we reviewed urological research works using microfluidic devices. Microfluidic devices were used for the detection of prostate and bladder cancer and the characterization of cancer microenvironments. The potential applications of microfluidics in urinary analysis and sperm sorting were demonstrated. The use of microfluidic devices in urology research can provide high-throughput, high-precision, and low-cost analyzing platforms.

Differentiation Potential of Mesenchymal Stem Cells Is Related to Their Intrinsic Mechanical Properties.

PURPOSE: The differentiation properties of stem cells are not yet fully understood due to their close association with multiple environmental and extrinsic factors. This study investigates the differentiation properties of mesenchymal stem cells (MSCs) and correlates them with their intrinsic mechanical properties. METHODS: A total of 3 different types of MSCs, namely bone marrow-derived MSCs (BMSCs), umbilical cord-derived MSCs (UCSCs), and adipose-derived MSCs (ADSCs) were evaluated. These 3 MSCs were individually differentiated into adipocytes and osteoblasts for 3 weeks. The mechanical properties of the MSCs and differentiated cells were determined by atomic force microscopy. RESULTS: ADSCs showed the greatest ability to differentiate into adipocytes, followed by BMSCs and UCSCs. While UCSCs differentiated readily into osteoblasts, BMSCs and ADSCs were less likely to undergo this differentiation. UCSCs were the "hardest" cells, while ADSCs were the "softest." The cells differentiated from "hard" MSCs were stiffer than the cells differentiated from "soft" MSCs, irrespective of lineage specification. CONCLUSIONS: The differentiation ability of MSCs and the mechanical properties of the differentiated cells were closely linked. However, there were no significant correlations regarding changes in the mechanical properties between the nuclear region and the cytoplasm during differentiation.

Colorimetric detection of endogenous hydrogen sulfide production in living cells.

Hydrogen sulfide (H2S) has received great attention as a third gaseous signal transmitter, following nitric oxide and carbon monoxide. In particular, H2S plays an important role in the regulation of cancer cell biology. Therefore, the detection of endogenous H2S concentrations within biological systems can be helpful to understand the role of gasotransmitters in pathophysiology. Although a simple and inexpensive method for the detection of H2S has been developed, its direct and precise measurement in living cells remains a challenge. In this study, we introduced a simple, facile, and inexpensive colorimetric system for selective H2S detection in living cells using a silver-embedded Nafion/polyvinylpyrrolidone (PVP) membrane. This membrane could be easily applied onto a polystyrene microplate cover. First, we optimized the composition of the coating membrane, such as the PVP/Nafion mixing ratio and AgNO3 concentration, as well as the pH of the Na2S (H2S donor) solution and the reaction time. Next, the in vitro performance of a colorimetric detection assay utilizing the silver/Nafion/PVP membrane was evaluated utilizing a known concentration of Na2S standard solution both at room temperature and at 37°C in a 5% CO2 incubator. As a result, the sensitivity of the colorimetric assay for H2S at 37°C in the incubator (0.0056Abs./µM Na2S, R2=0.9948) was similar to that at room temperature (0.0055Abs./µM Na2S, R2=0.9967). Moreover, these assays were less sensitive to interference from compounds such as glutathione, l-cysteine (Cys), and dithiothreitol than to the H2S from Na2S. This assay based on the silver/Nafion/PVP membrane also showed excellent reproducibility (2.8% RSD). Finally, we successfully measured the endogenous H2S concentrations in live C6 glioma cells by s-(5'-adenosyl)-l-methionine stimulation with and without Cys and l-homocysteine, utilizing the silver/Nafion/PVP membrane. In summary, colorimetric assays using silver/Nafion/PVP-coated membranes can be simple, robust, and reliable tools for the detection of H2S that can avoid the complicated and labor-intensive analytical approach used in conventional biology. In addition, we expect that this assay will demonstrate a powerful ability to study pathophysiological pathways that involve H2S.

Controlling successive ionic layer absorption and reaction cycles to optimize silver nanoparticle-induced localized surface plasmon resonance effects on the paper strip.

This study investigates why a silver nanoparticle (SNP)-induced surface-enhanced Raman scattering (SERS) paper chip fabricated at low successive ionic layer absorption and reaction (SILAR) cycles leads to a high SERS enhancement factor (7×108) with an inferior nanostructure and without generating a hot spot effect. The multi-layered structure of SNPs on cellulose fibers, verified by magnified scanning electron microscopy (SEM) and analyzed by a computational simulation method, was hypothesized as the reason. The pattern of simulated local electric field distribution with respect to the number of SILAR cycles showed good agreement with the experimental Raman intensity, regardless of the wavelength of the excitation laser sources. The simulated enhancement factor at the 785-nm excitation laser source (2.8×109) was 2.5 times greater than the experimental enhancement factor (1.1×109). A 532-nm excitation laser source exhibited the highest maximum local electric field intensity (1.9×1011), particularly at the interparticle gap called a hot spot. The short wavelength led to a strong electric field intensity caused by strong electromagnetic coupling arising from the SNP-induced local surface plasmon resonance (LSPR) effects through high excitation energy. These findings suggest that our paper-based SILAR-fabricated SNP-induced LSPR model is valid for understanding SNP-induced LSPR effects.

Exenatide Prevents Morphological and Structural Changes of Mitochondria Following Ischaemia-Reperfusion Injury.

BACKGROUND: Exenatide exerts cardioprotective effects by attenuating ischaemic reperfusion (IR) injury, possibly through activating the opening of mitochondrial ATP-sensitive potassium channels. We used atomic force microscopy (AFM) to investigate changes in mitochondrial morphology and properties in order to assess exenatide-mediated cardioprotection in IR injury. METHODS: We used an in vivo Sprague-Dawley rat IR model and ex vivo Langendorff injury model. In the left anterior descending artery (LAD) occlusion model, animals were randomly divided into three groups: sham-operated rats (Sham, n=5), IR-injured rats treated with placebo (IR, n=6), and IR-injured treated with exenatide (IR + EXE, n=6). For the Langendorff model, rats were randomly divided into two groups: IR injury with placebo (IR, n=4) and IR injury with exenatide (IR+EXE, n=4). Morphological and mechanical changes of mitochondria were analysed by AFM. RESULTS: Exenatide pre-treatment improved cardiac function as evidenced by improvement in echocardiographic results. The ratio of infarct area (IA) to risk area (RA) was significantly reduced in exenatide-treated rats. According to AFM, IR significantly increased the area of isolated mitochondria, indicative of mitochondrial swelling. Treatment with exenatide reduced the mitochondrial area and ameliorated the adhesion force of mitochondrial surfaces. CONCLUSIONS: Exenatide pre-treatment improves morphological and mechanical characteristics of mitochondria in response to IR injury in a rat model. These alterations in mitochondrial characteristics appear to play a cardioprotective role against IR injury.

Changes in nail keratin observed by Raman spectroscopy after Nd:YAG laser treatment.

Lasers and photodynamic therapy have been considered a convergence treatment for onychomycosis, which is a fungal infection on the nail bed and nail plate. Laser therapies have shown satisfactory results without significant complications for onychomycosis; however, the mechanism of clearing remains unknown. In this work, we investigated changes in the chemical structure of nail keratin induced by Nd:YAG laser using Raman spectroscopy. Toe nails with onychomycosis were treated with 1064 nm Nd:YAG laser. After laser treatment, the disulfide band (490-590 cm-1 ) of nail keratin was rarely observed or was reduced in intensity. The amide I band (1500-1700 cm-1 ) also showed changes induced by the laser. The α-helical (1652 cm-1 ) structures dominated the ß-sheet (1673 cm-1 ) in nontreated nail, but the opposite phenomenon was observed after laser treatment.

Hemorheological characteristics of red blood cells exposed to surface functionalized graphene quantum dots.

Graphene quantum dots (GQDs) are potential candidates for various biomedical applications such as drug delivery, bioimaging, cell labeling, and biosensors. However, toxicological information on their effects on red blood cells (RBCs) and the mechanisms involved remain unexplored. To the best of our knowledge, our study is the first to investigate the toxicity effects of three GQDs with different surface functionalizations on the hemorheological characteristics of human RBCs, including hemolysis, deformability, aggregation, and morphological changes. RBCs were exposed to three different forms of GQDs (non-functionalized, hydroxylated, and carboxylated GQDs) at various concentrations (0, 500, 750, and 1000 µg/mL) and incubation times (0, 1, 2, 3, or 4 h). The rheological characteristics of the RBCs were measured using microfluidic-laser diffractometry and aggregometry. Overall, the hemolysis rate and rheological alterations of the RBCs were insignificant at a concentration less than 500 µg/mL. Carboxylated GQDs were observed to have more substantial hemolytic activity and caused abrupt changes in the deformability and aggregation of the RBCs than the non-functionalized or hydroxylated GQDs at concentrations >750 µg/mL. Our findings indicate that hemorheological assessments could be utilized to estimate the degree of toxicity to cells and to obtain useful information on safety sheets for nanomaterials.

Selective cytotoxic effect of non-thermal micro-DBD plasma.

Non-thermal plasma has been extensively researched as a new cancer treatment technology. We investigated the selective cytotoxic effects of non-thermal micro-dielectric barrier discharge (micro-DBD) plasma in cervical cancer cells. Two human cervical cancer cell lines (HeLa and SiHa) and one human fibroblast (HFB) cell line were treated with micro-DBD plasma. All cells underwent apoptotic death induced by plasma in a dose-dependent manner. The plasma showed selective inhibition of cell proliferation in cervical cancer cells compared to HFBs. The selective effects of the plasma were also observed between the different cervical cancer cell lines. Plasma treatment significantly inhibited the proliferation of SiHa cells in comparison to HeLa cells. The changes in gene expression were significant in the cervical cancer cells in comparison to HFBs. Among the cancer cells, apoptosis-related genes were significantly enriched in SiHa cells. These changes were consistent with the differential cytotoxic effects observed in different cell lines.

Biochemical fingerprints of human papillomavirus infection and cervical dysplasia using cervical fluids: Spectral pattern investigation.

The Pap smear is the primary screening tool for invasive cervical cancer resulting from a persistent infection with oncogenic human papillomavirus (HPV); however, there are the problems such as the inability to distinguish between HPV infection and cervical dysplasia and a low sensitivity remain. We present preliminary findings of a label-free method to detect and classify HPV infection and cervical dysplasia using human cervical fluids. Three experimental groups, defined as normal, HPV-positive, and cervical dysplasia, were evaluated through their Raman spectral patterns for noise-independence, high reproducibility, and uniformity. Clinical diagnosis was performed through liquid-based cervical cytology, HPV test, and cervical histologic examination. Healthy cervical fluids showed a strong Raman intensity at 877 cm-1 (symmetric C-C stretching), and at 963 cm-1 (phosphate), compared to a reference Raman peak at 1003 cm-1 (phenylalanine symmetric ring breath). The HPV-positive cervical fluids showed a strong intensity of a Raman peak at 1448 cm-1 corresponding to C-H deformation vibration mode and the highest similarity between the central and ring zones among the three groups. The cervical dysplasia fluids showed the presence of strong peaks compared to the control and HPV-positive groups. In addition, different Raman spectra were acquired according to HPV type. Therefore, all ranges of cervical fluid-induced Raman spectra could be used to detect the presence of cervical pre-cancer. Raman peak-gated assessment provides a label-free and nondestructive tool for the clinical diagnosis of HPV infection and cervical precancerous changes.

A solvent-free microbial-activated air cathode battery paper platform made with pencil-traced graphite electrodes.

We present the fabrication of an ultra-low cost, disposable, solvent-free air cathode all-paper microbial fuel cell (MFC) that does not utilize any chemical treatments. The anode and cathode were fabricated by depositing graphite particles by drawing them on paper with a pencil (four strokes). Hydrophobic parchment paper was used as a proton exchange membrane (PEM) to allow only H(+) to pass. Air cathode MFC technology, where O2 was used as an electron acceptor, was implemented on the paper platform. The bioelectric current was generated by an electrochemical process involving the redox couple of microbial-activated extracellular electron transferred electrons, PEM-passed H(+), and O2 in the cathode. A fully micro-integrated pencil-traced MFC showed a fast start-time, producing current within 10 s after injection of bacterial cells. A single miniaturized all-paper air cathode MFC generated a maximum potential of 300 mV and a maximum current of 11 µA during 100 min after a single injection of Shewanella oneidensis. The micro-fabricated solvent-free air cathode all-paper MFC generated a power of 2,270 nW (5.68 mW/m(2)). The proposed solvent-free air cathode paper-based MFC device could be used for environmentally-friendly energy storage as well as in single-use medical power supplies that use organic matter.

Enhancement of Matrix Metalloproteinase-2 (MMP-2) as a Potential Chondrogenic Marker during Chondrogenic Differentiation of Human Adipose-Derived Stem Cells.

Human adipose-derived stem cells (hASCs) have a capacity to undergo adipogenic, chondrogenic, and osteogenic differentiation. Recently, hASCs were applied to various fields including cell therapy for tissue regeneration. However, it is hard to predict the direction of differentiation of hASCs in real-time. Matrix metalloproteinases (MMPs) are one family of proteolytic enzymes that plays a pivotal role in regulating the biology of stem cells. MMPs secreted by hASCs are expected to show different expression patterns depending on the differentiation state of hASCs because biological functions exhibit different patterns during the differentiation of stem cells. Here, we investigated proteolytic enzyme activity, especially MMP-2 activity, in hASCs during their differentiation. The activities of proteolytic enzymes and MMP-2 were higher during chondrogenic differentiation than during adipogenic and osteogenic differentiation. During chondrogenic differentiation, mRNA expression of MMP-2 and the level of the active form of MMP-2 were increased, which also correlated with Col II. It is concluded that proteolytic enzyme activity and the level of the active form of MMP-2 were increased during chondrogenic differentiation, which was accelerated in the presence of Col II protein. According to our findings, MMP-2 could be a candidate maker for real-time detection of chondrogenic differentiation of hASCs.

Instrument-Free Synthesizable Fabrication of Label-Free Optical Biosensing Paper Strips for the Early Detection of Infectious Keratoconjunctivitides.

We introduce a surface-enhanced Raman scattering (SERS)-functionalized, gold nanoparticle (GNP)-deposited paper strip capable of label-free biofluid sensing for the early detection of infectious eye diseases. The GNP biosensing paper strip was fabricated by the direct synthesis and deposition of GNPs on wax-divided hydrophilic areas of a permeable porous substrate through a facile, power-free synthesizable, and highly reproducible successive ionic layer absorption and reaction (SILAR) technique. To maximize localized surface plasmon resonance-generated SERS activity, the concentration of the reactive solution and number of SILAR cycles were optimized by controlling the size and gap distance of GNPs and verified by computational modeling with geometrical hypotheses of Gaussian-estimated metallic nanoparticles. The responses of our SERS-functionalized GNP paper strip to Raman intensities exhibited an enhancement factor of 7.8 × 10(8), high reproducibility (relative standard deviation of 7.5%), and 1 pM 2-naphthalenethiol highly sensitive detection limit with a correlation coefficient of 0.99, achieved by optimized SILAR conditions including a 10/10 mM/mM HAuCl4/NaBH4 concentration and six SILAR cycles. The SERS-functionalized GNP paper is supported by a multivariate statistics-preprocessed machine learning-judged bioclassification system to provide excellent label-free chemical structure sensitivity for identifying infectious keratoconjunctivitis. The power-free synthesizable fabrication, label-free, rapid analysis, and high sensitivity feature of the SILAR-fabricated SERS-functionalized GNP biosensing paper strip makes it an excellent alternative in point-of-care applications for the early detection of various infectious diseases.