Dynamic Measurement of a Cancer Biomarker: Towards In Situ Application of a Fiber-Optic Ball Resonator Biosensor in CD44 Protein Detection.

The accuracy and efficacy of medical treatment would be greatly improved by the continuous and real-time monitoring of protein biomarkers. Identification of cancer biomarkers in patients with solid malignant tumors is receiving increasing attention. Existing techniques for detecting cancer proteins, such as the enzyme-linked immunosorbent assay, require a lot of work, are not multiplexed, and only allow for single-time point observations. In order to get one step closer to clinical usage, a dynamic platform for biosensing the cancer biomarker CD44 using a single-mode optical fiber-based ball resonator biosensor was designed, constructed and evaluated in this work. The main novelty of the work is an in-depth study of the capability of an in-house fabricated optical fiber biosensor for in situ detection of a cancer biomarker (CD44 protein) by conducting several types of experiments. The main results of the work are as follows: (1) Calibration of the fabricated fiber-optic ball resonator sensors in both static and dynamic conditions showed similar sensitivity to the refractive index change demonstrating its usefulness as a biosensing platform for dynamic measurements; (2) The fabricated sensors were shown to be insensitive to pressure changes further confirming their utility as an in situ sensor; (3) The sensor's packaging and placement were optimized to create a better environment for the fabricated ball resonator's performance in blood-mimicking environment; (4) Incubating increasing protein concentrations with antibody-functionalized sensor resulted in nearly instantaneous signal change indicating a femtomolar detection limit in a dynamic range from 7.1 aM to 16.7 nM; (5) The consistency of the obtained signal change was confirmed by repeatability studies; (6) Specificity experiments conducted under dynamic conditions demonstrated that the biosensors are highly selective to the targeted protein; (7) Surface morphology studies by AFM measurements further confirm the biosensor's exceptional sensitivity by revealing a considerable shift in height but no change in surface roughness after detection. The biosensor's ability to analyze clinically relevant proteins in real time with high sensitivity offers an advancement in the detection and monitoring of malignant tumors, hence improving patient diagnosis and health status surveillance.

Optical Fiber Distributed Sensing Network for Thermal Mapping in Radiofrequency Ablation Neighboring a Blood Vessel.

Radiofrequency ablation (RFA) is a minimally invasive form of thermotherapy with great potential in cancer care, having the capability of selectively ablating tumoral masses with a surface area of several cm2. When performing RFA in the proximity of a blood vessel, the heating profile changes due to heat dissipation, perfusion, and impedance changes. In this work, we provide an experimental framework for the real-time evaluation of 2D thermal maps in RFA neighboring a blood vessel; the experimental setup is based on simultaneous scanning of multiple fibers in a distributed sensing network, achieving a spatial resolution of 2.5 × 4 mm2 in situ. We also demonstrate an increase of ablating potential when injecting an agarose gel in the tissue. Experimental results show that the heat-sink effect contributes to a reduction of the ablated region around 30-60% on average; however, the use of agarose significantly mitigates this effect, enlarging the ablated area by a significant amount, and ablating an even larger surface (+15%) in the absence of blood vessels.

Spatial-Division Multiplexing Approach for Simultaneous Detection of Fiber-Optic Ball Resonator Sensors: Applications for Refractometers and Biosensors.

Fiber-optic ball resonators are an attractive technology for refractive index (RI) sensing and optical biosensing, as they have good sensitivity and allow for a rapid and repeatable manufacturing process. An important feature for modern biosensing devices is the multiplexing capacity, which allows for interrogating multiple sensors (potentially, with different functionalization methods) simultaneously, by a single analyzer. In this work, we report a multiplexing method for ball resonators, which is based on a spatial-division multiplexing approach. The method is validated on four ball resonator devices, experimentally evaluating both the cross-talk and the spectral shape influence of one sensor on another. We show that the multiplexing approach is highly efficient and that a sensing network with an arbitrary number of ball resonators can be designed with reasonable penalties for the sensing capabilities. Furthermore, we validate this concept in a four-sensor multiplexing configuration, for the simultaneous detection of two different cancer biomarkers across a widespread range of concentrations.

Highly Sensitive Zinc Oxide Fiber-Optic Biosensor for the Detection of CD44 Protein.

Currently, significant progress is being made in the prevention, treatment and prognosis of many types of cancer, using biological markers to assess current physiological processes in the body, including risk assessment, differential diagnosis, screening, treatment determination and monitoring of disease progression. The interaction of protein coding gene CD44 with the corresponding ligands promotes the processes of invasion and migration in metastases. The study of new and rapid methods for the quantitative determination of the CD44 protein is essential for timely diagnosis and therapy. Current methods for detecting this protein use labeled assay reagents and are time consuming. In this paper, a fiber-optic biosensor with a spherical tip coated with a thin layer of zinc oxide (ZnO) with a thickness of 100 nm, deposited using a low-cost sol-gel method, is developed to measure the CD44 protein in the range from 100 aM to 100 nM. This sensor is easy to manufacture, has a good response to the protein change with detection limit of 0.8 fM, and has high sensitivity to the changes in the refractive index (RI) of the environment. In addition, this work demonstrates the possibility of achieving sensor regeneration without damage to the functionalized surface. The sensitivity of the obtained sensor was tested in relation to the concentration of the control protein, as well as without antibodies-CD44.

Fiber-Optic Distributed Sensing Network for Thermal Mapping of Gold Nanoparticles-Mediated Radiofrequency Ablation.

In this work, we report the design of an optical fiber distributed sensing network for the 2-dimensional (2D) in situ thermal mapping of advanced methods for radiofrequency thermal ablation. The sensing system is based on six high-scattering MgO-doped optical fibers, interleaved by a scattering-level spatial multiplexing approach that allows simultaneous detection of each fiber location, in a 40 × 20 mm grid (7.8 mm2 pixel size). Radiofrequency ablation (RFA) was performed on bovine phantom, using a pristine approach and methods mediated by agarose and gold nanoparticles in order to enhance the ablation properties. The 2D sensors allow the detection of spatiotemporal patterns, evaluating the heating properties and investigating the repeatability. We observe that agarose-based ablation yields the widest ablated area in the best-case scenario, while gold nanoparticles-mediated ablation provides the best trade-off between the ablated area (53.0-65.1 mm2, 61.5 mm2 mean value) and repeatability.

Ultra-wide, attomolar-level limit detection of CD44 biomarker with a silanized optical fiber biosensor.

Measuring cancer biomarkers at ultralow detection limit and high sensitivity could be a promising tool for early diagnosis, monitoring treatment and post-treatment recurrence. Soluble CD44 is a promising diagnostic and prognostic biomarker in several types of cancer including gastric, colon and breast cancer. Several highly sensitive biosensors have been built to measure this important biomarker. However, they did not reach attomolar level of detection. The aim of this work was to build a biosensor capable of detecting CD44 concentrations down to attomolar (aM) level while measuring it in a wide concentration range. Herein, we demonstrate a biosensor that offers 4 key advantages over existing platforms for CD44 detection: 1) detection of CD44 was carried out in a diluted serum down to attomolar level (4.68 aM) which is about 6 orders of magnitude lower than that of a traditional ELISA; 2) fabrication of the sensor is done in a fast way using inexpensive materials making it a disposable fiber optic biosensor; 3) detection of CD44 was performed in a wide dynamic range previously not shown in other similar biosensors; 4) a proof-of-concept experiment was performed using the biosensor to embed it in a catheter to measure the protein in flow conditions.

Green-Synthesized Silver Nanoparticle-Assisted Radiofrequency Ablation for Improved Thermal Treatment Distribution.

Thermal ablation therapy is known as an advantageous alternative to surgery allowing the treatment of multiple tumors located in hard-to-reach locations or treating patients with medical conditions that are not compatible with surgery. Appropriate heat propagation and precise control over the heat propagation is considered a weak point of thermal ablation therapy. In this work, silver nanoparticles (AgNPs) are used to improve the heat propagation properties during the thermal ablation procedure. Green-synthesized silver nanoparticles offer several attractive features, such as excellent thermal conductivity, biocompatibility, and antimicrobial activity. A distributed multiplexed fiber optic sensing system is used to monitor precisely the temperature change during nanoparticle-assisted radiofrequency ablation. An array of six MgO-based nanoparticles doped optical fibers spliced to single-mode fibers allowed us to obtain the two-dimensional thermal maps in a real time employing optical backscattering reflectometry at 2 mm resolution and 120 sensing points. The silver nanoparticles at 5, 10, and 20 mg/mL were employed to investigate their heating effects at several positions on the tissue regarding the active electrode. In addition, the pristine tissue and tissue treated with agarose solution were also tested for reference purposes. The results demonstrated that silver nanoparticles could increase the temperature during thermal therapies by propagating the heat. The highest temperature increase was obtained for 5 mg/mL silver nanoparticles introduced to the area close to the electrode with a 102% increase of the ablated area compared to the pristine tissue.

Thermo-Visco-Elastometry of RF-Wave-Heated and Ablated Flesh Tissues Containing Au Nanoparticles.

We report non-contact laser-based Brillouin light-scattering (BLS) spectroscopy measurements of the viscoelastic properties of hyperthermally radiofrequency (RF)-heated and ablated bovine liver and chicken flesh tissues with embedded gold nanoparticles (AuNPs). The spatial lateral profile of the local surface temperature in the flesh samples during their hyperthermia was measured through optical backscattering reflectometry (OBR) using Mg−silica-NP-doped sensing fibers distributed with an RF applicator and correlated with viscoelastic variations in heat-affected and ablated tissues. Substantial changes in the tissue stiffness after heating and ablation were directly related to their heat-induced structural modifications. The main proteins responsible for muscle elasticity were denatured and irreversibly aggregated during the RF ablation. At T > 100 °C, the proteins constituting the flesh further shrank and became disorganized, leading to substantial plastic deformation of biotissues. Their uniform destruction with larger thermal lesions and a more viscoelastic network was attained via AuNP-mediated RF hyperthermal ablation. The results demonstrated here pave the way for simultaneous real-time hybrid optical sensing of viscoelasticity and local temperature in biotissues during their denaturation and gelation during hyperthermia for future applications that involve mechanical- and thermal-property-controlled theranostics.

Label-free fiber-optic spherical tip biosensor to enable picomolar-level detection of CD44 protein.

Increased level of CD44 protein in serum is observed in several cancers and is associated with tumor burden and metastasis. Current clinically used detection methods of this protein are time-consuming and use labeled reagents for analysis. Therefore exploring new label-free and fast methods for its quantification including its detection in situ is of importance. This study reports the first optical fiber biosensor for CD44 protein detection, based on a spherical fiber optic tip device. The sensor is easily fabricated from an inexpensive material (single-mode fiber widely used in telecommunication) in a fast and robust manner through a CO2 laser splicer. The fabricated sensor responded to refractive index change with a sensitivity of 95.76 dB/RIU. The spherical tip was further functionalized with anti-CD44 antibodies to develop a biosensor and each step of functionalization was verified by an atomic force microscope. The biosensor detected a target of interest with an achieved limit of detection of 17 pM with only minor signal change to two control proteins. Most importantly, concentrations tested in this work are very broad and are within the clinically relevant concentration range. Moreover, the configuration of the proposed biosensor allows its potential incorporation into an in situ system for quantitative detection of this biomarker in a clinical setting.

Optical Fiber Ball Resonator Sensor Spectral Interrogation through Undersampled KLT: Application to Refractive Index Sensing and Cancer Biomarker Biosensing.

Optical fiber ball resonators based on single-mode fibers in the infrared range are an emerging technology for refractive index sensing and biosensing. These devices are easy and rapid to fabricate using a CO2 laser splicer and yield a very low finesse reflection spectrum with a quasi-random pattern. In addition, they can be functionalized for biosensing by using a thin-film sputtering method. A common problem of this type of device is that the spectral response is substantially unknown, and poorly correlated with the size and shape of the spherical device. In this work, we propose a detection method based on Karhunen-Loeve transform (KLT), applied to the undersampled spectrum measured by an optical backscatter reflectometer. We show that this method correctly detects the response of the ball resonator in any working condition, without prior knowledge of the sensor under interrogation. First, this method for refractive index sensing of a gold-coated resonator is applied, showing 1594 RIU-1 sensitivity; then, this concept is extended to a biofunctionalized ball resonator, detecting CD44 cancer biomarker concentration with a picomolar-level limit of detection (19.7 pM) and high specificity (30-41%).

Distributed Sensing Network Enabled by High-Scattering MgO-Doped Optical Fibers for 3D Temperature Monitoring of Thermal Ablation in Liver Phantom.

Thermal ablation is achieved by delivering heat directly to tissue through a minimally invasive applicator. The therapy requires a temperature control between 50-100 °C since the mortality of the tumor is directly connected with the thermal dosimetry. Existing temperature monitoring techniques have limitations such as single-point monitoring, require costly equipment, and expose patients to X-ray radiation. Therefore, it is important to explore an alternative sensing solution, which can accurately monitor temperature over the whole ablated region. The work aims to propose a distributed fiber optic sensor as a potential candidate for this application due to the small size, high resolution, bio-compatibility, and temperature sensitivity of the optical fibers. The working principle is based on spatial multiplexing of optical fibers to achieve 3D temperature monitoring. The multiplexing is achieved by high-scattering, nanoparticle-doped fibers as sensing fibers, which are spatially separated by lower-scattering level of single-mode fibers. The setup, consisting of twelve sensing fibers, monitors tissue of 16 mm × 16 mm × 25 mm in size exposed to a gold nanoparticle-mediated microwave ablation. The results provide real-time 3D thermal maps of the whole ablated region with a high resolution. The setup allows for identification of the asymmetry in the temperature distribution over the tissue and adjustment of the applicator to follow the allowed temperature limits.

Distributed 2D temperature sensing during nanoparticles assisted laser ablation by means of high-scattering fiber sensors.

The high demand in effective and minimally invasive cancer treatments, namely thermal ablation, leads to the demand for real-time multi-dimensional thermometry to evaluate the treatment effectiveness, which can be also assisted by the use of nanoparticles. We report the results of 20-nm gold and magnetic iron oxide nanoparticles-assisted laser ablation on a porcine liver phantom. The experimental set-up consisting of high-scattering nanoparticle-doped fibers was operated by means of a scattering-level multiplexing arrangement and interrogated via optical backscattered reflectometry, together with a solid-state laser diode operating at 980 nm. The multiplexed 2-dimensional fiber arrangement based on nanoparticle-doped fibers allowed an accurate superficial thermal map detected in real-time.

Application of Nanoparticles and Nanomaterials in Thermal Ablation Therapy of Cancer.

Cancer is one of the major health issues with increasing incidence worldwide. In spite of the existing conventional cancer treatment techniques, the cases of cancer diagnosis and death rates are rising year by year. Thus, new approaches are required to advance the traditional ways of cancer therapy. Currently, nanomedicine, employing nanoparticles and nanocomposites, offers great promise and new opportunities to increase the efficacy of cancer treatment in combination with thermal therapy. Nanomaterials can generate and specifically enhance the heating capacity at the tumor region due to optical and magnetic properties. The mentioned unique properties of nanomaterials allow inducing the heat and destroying the cancerous cells. This paper provides an overview of the utilization of nanoparticles and nanomaterials such as magnetic iron oxide nanoparticles, nanorods, nanoshells, nanocomposites, carbon nanotubes, and other nanoparticles in the thermal ablation of tumors, demonstrating their advantages over the conventional heating methods.

Multi-fiber distributed thermal profiling of minimally invasive thermal ablation with scattering-level multiplexing in MgO-doped fibers.

We propose a setup for multiplexed distributed optical fiber sensors capable of resolving temperature distribution in thermo-therapies, with a spatial resolution of 2.5 mm over multiple fibers interrogated simultaneously. The setup is based on optical backscatter reflectometry (OBR) applied to optical fibers having backscattered power significantly larger than standard fibers (36.5 dB), obtained through MgO doping. The setup is based on a scattering-level multiplexing, which allows interrogating all the sensing fibers simultaneously, thanks to the fact that the backscattered power can be unambiguously associated to each fiber. The setup has been validated for the planar measurement of temperature profiles in ex vivo radiofrequency ablation, obtaining the measurement of temperature over a surface of 96 total points (4 fibers, 8 sensing points per cm2). The spatial resolution obtained for the planar measurement allows extending distributed sensing to surface, or even three-dimensional, geometries performing temperature sensing in the tissue with millimeter resolution in multiple dimensions.