All-fiber label-free optical fiber biosensors: from modern technologies to current applications [Invited].

Biosensors are established as promising analytical tools for detecting various analytes important in biomedicine and environmental monitoring. Using fiber optic technology as a sensing element in biosensors offers low cost, high sensitivity, chemical inertness, and immunity to electromagnetic interference. Optical fiber sensors can be used in in vivo applications and multiplexed to detect several targets simultaneously. Certain configurations of optical fiber technology allow the detection of analytes in a label-free manner. This review aims to discuss recent advances in label-free optical fiber biosensors from a technological and application standpoint. First, modern technologies used to build label-free optical fiber-based sensors will be discussed. Then, current applications where these technologies are applied are elucidated. Namely, examples of detecting soluble cancer biomarkers, hormones, viruses, bacteria, and cells are presented.

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.

Functionalized optical fiber ball-shaped biosensor for label-free, low-limit detection of IL-8 protein.

Detection of biomarkers for tracking disease progression is becoming increasingly important in biomedicine. Using saliva as a diagnostic sample appears to be a safe, cost-effective, and non-invasive approach. Salivary interleukin-8 levels demonstrate specific changes associated with diseases such as obstructive pulmonary disease, squamous cell carcinoma, oral cancer, and breast cancer. Traditional protein detection methods, such as enzyme-linked immunosorbent assay (ELISA), mass spectrometry, and Western blot are often expensive, complex, and time-consuming. In this study, an optical fiber-based biosensor was developed to detect salivary IL-8 protein in a label-free manner. The biosensor was able to achieve an ultra-low limit detection of 0.91 fM. Moreover, the tested concentration range was wide: from 273 aM to 59 fM. As a proof-of-concept for detecting the protein in real clinical samples, the detection was carried out in artificial saliva. It was possible to achieve high sensitivity for the target protein and minimal signal alterations for the control proteins.

Reversed distal laterodigital adipofascial flap for nail-bed reconstruction.

BACKGROUND: Lesions of the distal phalanges of the fingers frequently involve the nail bed. There are few therapeutic options for nail-bed reconstruction and they often lead to painful scars and onychodystrophy. We present our experience with the distal adipofascial laterodigital reverse flap. METHODS: Fifteen patients (average age 46.33 years, range 28-73) with tumors or traumatic injuries (crush injuries, nail avulsion, and partial fingertip amputations) of the nail bed, underwent digital reconstruction through the distal adipofascial laterodigital reverse flap from June 2018 to August 2019. The size of the fingertip defect covered with the flap was ranged between 1.1 × 1.1 and 1.6 × 1.2 cm (average size 1.4 × 1.2 cm). The flap was harvested enrolling subcutaneous tissue from the lateral aspect of the middle and distal phalanx from the less damaged side. RESULTS: The average size of the harvested flaps was 1.3 × 1.2 cm (range 1.1 × 1.0 to 1.4 × 1.1 cm). All adipofascial flaps survived entirely and the nail bed healed in all patients, with an average healing time of 21 days and a subsequent regrowth of the nail. The follow up ranged from 6 to 12 months, with a mean of 7 months. CONCLUSIONS: The distal reverse adipofascial flap provides a very versatile and reliable coverage of the distal finger and its nail bed. It is a rapid and reproducible surgical procedure with poor morbidity for the donor site. LEVEL OF EVIDENCE: IV.

Dorso-ulnar reverse flow pedicled osseous flap for reconstruction of the distal phalanx of the thumb: A case report.

Reconstruction of osseous defects of the distal phalanx of the thumb is usually addressed with free bone grafts or free vascularized bone flaps. Some reports demonstrated the possibility to harvest an osteo-cutaneous flap in the dorso-ulnar side of the first metacarpal bone with success. In the same manner, no reports are present in the literature in which bone deficits were reconstructed with this flap elevated as an exclusively osseous flap. We report our successful experience with one case of distal phalanx reconstruction of the thumb by mean of the dorso-ulnar reverse flow pedicled osseous flap. The patient was a 45-year-old woman with symptoms related to a cystic bone tumor that involved the entirety of the distal phalanx of the thumb. Flap dimensions were calculated based on x-ray gap measures, which resulted in need of 1.5 × 0.8 × 0.5 cm flap dimensions. An osseous flap was harvested and transposed from the ulnar side of the first metacarpal bone. K-wire fixation was utilized for bone flap stabilization. No complications occurred and excellent functional result was evaluated at 6 months follow-up. In our opinion, the flap may be considered as an alternative to free bone grafts in situations in which perilesional tissues may jeopardize the process of free graft taking and in cases in which free vascularized bone flaps are not feasible for patient or surgeon decision.

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.

Cost-Effective Fiber Optic Solutions for Biosensing.

In the last years, optical fiber sensors have proven to be a reliable and versatile biosensing tool. Optical fiber biosensors (OFBs) are analytical devices that use optical fibers as transducers, with the advantages of being easily coated and biofunctionalized, allowing the monitorization of all functionalization and detection in real-time, as well as being small in size and geometrically flexible, thus allowing device miniaturization and portability for point-of-care (POC) testing. Knowing the potential of such biosensing tools, this paper reviews the reported OFBs which are, at the moment, the most cost-effective. Different fiber configurations are highlighted, namely, end-face reflected, unclad, D- and U-shaped, tips, ball resonators, tapered, light-diffusing, and specialty fibers. Packaging techniques to enhance OFBs' application in the medical field, namely for implementing in subcutaneous, percutaneous, and endoscopic operations as well as in wearable structures, are presented and discussed. Interrogation approaches of OFBs using smartphones' hardware are a great way to obtain cost-effective sensing approaches. In this review paper, different architectures of such interrogation methods and their respective applications are presented. Finally, the application of OFBs in monitoring three crucial fields of human life and wellbeing are reported: detection of cancer biomarkers, detection of cardiovascular biomarkers, and environmental monitoring.

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.

Ultralow Limit Detection of Soluble HER2 Biomarker in Serum with a Fiber-Optic Ball-Tip Resonator Assisted by a Tilted FBG.

An optical-fiber biosensor has been developed for the detection of the breast cancer biomarker soluble human epidermal growth factor receptor-2 (sHER2). The sensor was fabricated by combining a tilted fiber Bragg grating (TFBG) with a ball resonator, allowing us to achieve an excellent sensitivity compared to other optical-fiber-based sensors. The sensor exhibits a resonance comb excited by the TFBG and the spectral profile of the ball resonator. The detection of sHER2 at extremely low concentrations was carried out by tracking the amplitude change of selected resonances. The therapeutic anti-HER2 monoclonal antibody Trastuzumab has been used to functionalize the biosensor with silane surface chemistry. The sensor features a sensitivity of 4034 dB/RIU with a limit of detection (LoD) in buffer and in a 1/10 diluted serum of 151.5 ag/mL and 3.7 pg/mL, respectively. At relatively high protein concentrations (64 ng/mL) binding to sHER (7.36 dB) as compared to control proteins (below 0.7 dB) attested the high specificity of sHER2 detection.

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%).

Vascularized bone grafts for post-traumatic defects in the upper extremity.

Vascularized bone grafts (VBGs) are widely employed to reconstruct upper extremity bone defects. Conventional bone grafting is generally used to treat defects smaller than 5-6 cm, when tissue vascularization is adequate and there is no infection risk. Vascularized fibular grafts (VFGs) are mainly used in the humerus, radius or ulna in cases of persistent non-union where traditional bone grafting has failed or for bone defects larger than 6 cm. Furthermore, VFGs are considered to be the standard treatment for large bone defects located in the radius, ulna and humerus and enable the reconstruction of soft-tissue loss, as VFGs can be harvested as osteocutaneous flaps. VBGs enable one-stage surgical reconstruction and are highly infection-resistant because of their autonomous vascularization. A vascularized medial femoral condyle (VFMC) free flap can be used to treat small defects and non-unions in the upper extremity. Relative contraindications to these procedures are diabetes, immunosuppression, chronic infections, alcohol, tobacco, drug abuse and obesity. The aim of our study was to illustrate the use of VFGs to treat large post-traumatic bone defects and osteomyelitis located in the upper extremity. Moreover, the use of VFMC autografts is presented.

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.

Fingertip defect reconstruction with a modified pivot flap.

The pivot flap can treat volar oblique defects of the fingers distal to the distal interphalangeal joint. We present our experience in 11 fingers (11 patients) using this flap, including some modifications that optimize flap harvesting, mobilization, inset, and vascular supply and outcomes. The flaps harvested had an average size of 2.4 × 2.0 cm. The patients were followed for an average of 21 months (range 12 to 36) after surgery. Sensitivity of the flap was evaluated with the Semmes-Weinstein monofilaments test and static two-point discrimination test. Cold intolerance was also evaluated. The pivot flap with the modifications demonstrates a reliable technique for fingertip defect reconstruction. The results, in terms of sensitivity and functional recovery, seem promising.Level of evidence: IV.

Optimizing Silanization to Functionalize Stainless Steel Wire: Towards Breast Cancer Stem Cell Isolation.

Chemically modified metal surfaces have been used to recognize and capture specific cell types and biomolecules. In this work, stainless steel wires were functionalized with aptamers against breast cancer stem cell markers. Stainless steel wires were first electropolished and silanized via electrodeposition. Aptamers were then attached to the silanized surface through a cross-linker. The functionalized wires were able to capture the target cells in an in vitro test. During surface modification steps, wires were analyzed by atomic force microscopy, cyclic voltammetry, scanning electron and fluorescence microscopy to determine their surface composition and morphology. Optimized conditions of silanization (applied potential, solution pH, heat treatment temperature) for obtaining an aptamer-functionalized wire were determined in this work together with the use of several surface characterization techniques suitable for small-sized and circular wires. These modified wires have potential applications for the in vivo capture of target cells in blood flow, since their small size allows their insertion as standard guidewires in biomedical devices.

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.