sensors

Review
Current and Emerging Technology for Continuous
Glucose Monitoring
Cheng Chen 1,† , Xue-Ling Zhao 1,† , Zhan-Hong Li 1 , Zhi-Gang Zhu 1, *, Shao-Hong Qian 2
and Andrew J. Flewitt 3
1 School of Environmental and Materials Engineering, College of Engineering,
Shanghai Polytechnic University, Shanghai 201209, China; [email protected] (C.C.);
[email protected] (X.-L.Z.); [email protected] (Z.-H.L.)
2 Department of Ophthalmology, Eye and ENT Hospital, Shanghai Medical College, Fudan University,
Shanghai 200231, China; [email protected]
3 Electrical Engineering Division, Department of Engineering, University of Cambridge, J J Thomson Avenue,
Cambridge CB3 0FA, UK; [email protected]
* Correspondence: [email protected]; Tel.: +86-21-5021-5021 (ext. 8325)
† These authors contributed equally to this work.

Academic Editors: Giovanni Sparacino, Andrea Facchinetti and J. Hans de Vries

Received: 14 October 2016; Accepted: 20 December 2016; Published: 19 January 2017

Abstract: Diabetes has become a leading cause of death worldwide. Although there is no cure for
diabetes, blood glucose monitoring combined with appropriate medication can enhance treatment
efficiency, alleviate the symptoms, as well as diminish the complications. For point-of-care purposes,
continuous glucose monitoring (CGM) devices are considered to be the best candidates for diabetes
therapy. This review focuses on current growth areas of CGM technologies, specifically focusing
on subcutaneous implantable electrochemical glucose sensors. The superiority of CGM systems is
introduced firstly, and then the strategies for fabrication of minimally-invasive and non-invasive
CGM biosensors are discussed, respectively. Finally, we briefly outline the current status and future
perspective for CGM systems.

Keywords: continuous glucose monitoring; glucose biosensor; implanted devices; mini-invasive;

non-invasive

1. Introduction
Diabetes mellitus is a worldwide epidemic disease affecting 422 million people, and is predicted
to be the seventh leading cause of death if the current morbidity trends continue [1]. Blood glucose
(BG) concentrations in diabetics can undulate significantly throughout a day, and they lead serious
consequences including kidney failure, strokes, heart attacks, high blood pressure, blindness and
coma [2,3]. BG levels could be finely controlled by insulin, and the abnormal concentration of BG is
caused by the absence of insulin secretion (type I diabetes, T1D) or defective insulin secretion and action
(type II diabetes, T2D). For maintaining BG within the euglycemic range, the BG concentration should
be detected at least twice a day and four times a day for T2D patients and T1D patients, respectively,
and combined therapies including drugs, exogenous insulin supply, diet and physical exercise [4–6].
The emergence of glucose sensors has provided patients the ability to self-monitor BG levels so as to
manage insulin levels, and thus control the mortality of diabetes mellitus. Traditional glucose detecting
devices primarily consist of glucose sensors based on electrochemical methods [7]. The frequent
analysis of BG levels requires a small blood sample (<1 µL) obtained by a “finger-pricking” collection
method, which is inconvenient and results in poor patient compliance. Such tests neglect nighttime
variations and might cause approximation of BG variations. Moreover, instantaneous monitoring

Sensors 2017, 17, 182; doi:10.3390/s17010182 www.mdpi.com/journal/sensors

Sensors 2017, 17, 182 2 of 19

sensors cannot provide real-time BG information, and thus are unable to warn of hypoglycemic
(low blood sugar, <3.0 mM) and hyperglycemic (high blood sugar, >11.1 mM) events in advance.
There are three generations in the development of glucose biosensors, the first-generation relied
on the use of the natural oxygen and the production-detection process of hydrogen peroxide, and the
second-generation sensors employ a non-physiological electron acceptor to shuttle electrons and thus
solve the oxygen deficiency. The market leading methods are based on second generation sensor
technology. The design of the third-generation glucose sensors aims to get rid of the leachable artificial
mediators and even the glucose enzyme. Therefore, continuous glucose monitoring (CGM) devices are
considered to be the ideal candidate for the next generation products to replace the currently used portable
glucose meters [8]. CGM reports BG levels as trends of glucose fluctuations during a day (including the
increasing or decreasing of BG), and these data could be utilized for various applications, including
retrospective analysis [9], hypo/hyperglycemia detection and prediction [10,11]. Ideally, intelligent
CGM devices could be linked to an insulin delivery pump to form an artificial pancreas [12], and the
benefit of CGM point-of-care tests for the self-management of diabetes patients is the reduced time
length spent in hypoglycemia and the increased time in euglycemia [13–15]. However, the current
gold standard glucose biosensors are invasive, and CGM systems still present a number of limitations,
such as biofouling, fibrous encapsulation of the implanted electrode, inflammation, and loss of host
vasculature, which seriously affect the precision and accuracy of the BG results [16]. Non-invasive
methods, on the other hand, are increasingly prevalent due to the highly sensitivity and better patient
compliance contrary to invasive ones [17]. Since there is a delay as glucose is transported from the
blood level to the interstitial fluid level, the real-time information detected from CGM sensors might
be 15–20 min later than BG, so the lag-of-time of interstitial glucose (IG) relative to BG limits the
result reliability in hypoglycemic emergency situations. Although, many enhancements have been
implemented to mitigate the inaccuracy of CGM, there is still lack of approval of CGM data for
adjusting insulin, since the dosage of insulin is strongly dependent on the BG level and rate of change
of BG. Another barrier is the lifetime of the sensors, as loss of function happens to most implantable
electrodes within 7 days, and the calibration would also create a series of problems such as cost, toxicity,
inconvenience and discomfort [18]. The integration of CGM with wireless devices could be an effective
method for developing closed loop systems like the Internet of Things. By 2017, there will be more
mobile phones around world than people, and the CGM data could be sent to the smart phones for
analysis to then trigger the release of drugs [19].
This review focuses particularly on the progress in the development of CGM technologies,
focusing specifically on subcutaneous implantable electrochemical glucose sensors, which are widely
studied and commercially available. We briefly introduce the superiority of CGM systems and discuss
the challenges in the development and implementation of CGM devices in Section 1. Strategies for the
fabrication of minimally-invasive CGM biosensors, including new materials, biocompatible coatings
and drug delivery systems, are presented in Section 2. The non-invasive technologies and concepts are
introduced in Section 3. Finally, we briefly outline the current status and future perspectives for CGM
systems in Section 4.

2. Mini-Invasive Biosensors for CGM

Dynamic glucose detection methods usually employ a tiny sensor inserted beneath the skin and
maintained for a certain period of time in contact with the interstitial fluid. Most minimally-invasive
systems for CGM are enzyme-based, whereas others are enzyme-free. For both enzyme and non-enzyme
methods, numerous efforts have been focused on the preparation, functionalization and modification of
the required electrodes [16,20,21].

2.1. Sensing Design and New Materials

Enzymatic electrodes use enzymes to catalyze reduction–oxidation (redox) reactions, and the
movement of electrons could thus produce a concentration-dependent current or voltage that
Sensors 2017, 17, 182 3 of 19

can be measured by electrodes. Frequently utilized electrodes for invasive sensor technology
include microdialysis, micropores, microneedles, subcutaneous amperometric electrodes, and
intravenous implantable devices. Currently, only microdialysis and subcutaneous devices have reached
commercialization. Microdialysis systems utilize a hollow microdialysis fiber which is perfused
with isotonic fluid from an ex vivo reservoir, and meanwhile IG freely diffuses into the fiber, and
then is pumped to an enzyme-based electrochemical sensor. This system has smaller dimensions
for implantation convenience, and the sensor is ex vivo, thus avoiding biofouling problems [22].
For subcutaneous methods, the design of the electrodes mainly aims at increasing the sensitivity,
selectivity and biocompatibility.
A major historical advance in the in vivo application of glucose biosensors was the classic
needle-type sensor, which generally consists of a platinum-iridium (Pt-Ir, φ 0.125 mm, Pt:Ir = 9:1))
wire working electrode with an immobilized mediator and enzyme on the surface, and/or a polymer
coating for subcutaneous implantation. A silver/silver chloride (Ag/AgCl) wire is wrapped around the
working electrode that serves as a counter electrode. Such devices are designed for human implantation
sensors that continuously operate for a few days and be replaced by the patient. Algorithm correction
for the transient differences (lag-time) between BG and IG concentrations have been investigated for
future applications. Kumetrix (Union City, CA, USA) has proposed a hand-held, battery-powered
electronic monitor with silicon micro-needles that are similar in size to a human hair. The aim is to
develop a meter system that accepts a cartridge loaded with disposable sampling devices. This action
will cause the micro-needle to penetrate the skin and draw a very small volume of blood (less than
100 nL).
Coil-type oxidase biosensors were developed based on the design of needle-type ones. Pt-Ir wire was
wound up along a 30-gauge needle to form a coil-shaped cylinder, which had a length of ca. 1 mm with
an outer diameter of 0.55 mm and an inner diameter of 0.3 mm. Such a design dramatically increased
the sensing area of flexible implantable biosensors within an acceptable length range and created a ca.
0.07 mm three inner storage chamber for extra enzyme immobilization. With modifications such as
etching of the surface of coil, hydrogel layer coatings and loading with certain drugs, the constant
response of the sensor could be maintained for at least 60 days [23].
As for commercialized glucose sensors, Abbott (Chicago, IL, USA) has developed a series of
BG monitoring products named FreeStyle since June 2000. A recent FreeStyle Libre device allows
patients to measure glucose within 1 s by scanning the sensor with a reader, and the sensor could work
for 14 days without calibration. As a result, the pain associated with testing is greatly reduced. In 2006,
the Dexcom (San Diego, CA, USA) SEVEN, the first real-time continuous glucose monitoring system,
was approved by the FDA. Recently the Dexcom G5, the first fully mobile CGM system, was launched.
Dynamic glucose data could be accessed safely by a compatible smart device. Another transdermal
implantable CGM system, the CT-100 (POCTech, Huzhou, China), utilizes a “parallel implant method”,
which offsets the limitation of the length of the sensor by taking full advantage of the subcutaneous
tissue and thus differs from Dexcom’s products. Such a design improves the sensitivity while
preventing misoperation causing the sensor to go into the muscle tissue.
The diagnosis and management of diabetes mellitus require a tight monitoring of glucose
concentration. An ideal sensor would be one that provides reliable real-time continuous monitoring of
glucose variations throughout the day with high selectivity and speed over extended periods under
harsh conditions. New sensing concepts, coupled with new materials and numerous technological
innovations have been made for enhancing the capabilities and improving the reliability of glucose
measuring devices.
The development of carbon nanotubes (CNTs) and graphene-based electrodes have been
intensively studied, and they show good safety when used in implantable electrodes for in vitro
testing [16]. With similar dimensions as redox proteins, carbon nanomaterials could be utilized as
effective electrical wiring/connectors with redox enzymes, which is one of the most promising materials
for enzymatic glucose biosensors. For bio-sensing applications, graphene and CNTs demonstrated
Sensors 2017, 17, 182 4 of 19
Sensors 2016, 16, 182 4 of 18

higher sensitivity
low working and faster
potentials. response
Our group has time
carried than
outtraditional
a comprehensiveelectrodes
study atofextremely
CVD-synthesizedlow working CNT
potentials.
fibers used as sensing electrodes to detect glucose solutions. The CNT fiber resembles an used
Our group has carried out a comprehensive study of CVD-synthesized CNT fibers as
electric
sensing electrodes
wire, relying to detect glucose
on nanoscale surface solutions.
topography TheandCNT fiber resembles
porosity, which can anfacilitate
electric wire, relying on
molecular-scale
nanoscale surface topography and porosity, which can facilitate molecular-scale
interactions with agents like enzymes to efficiently capture and promote electron transfer reactions interactions with
agents like enzymes to efficiently capture and promote electron transfer
[24,25]. A scalable synthesis of a multifunctional conducting polyacrylic acid (PAA) hydrogel for reactions [24,25]. A scalable
synthesis of a multifunctional
glucose detection that integrates conducting polyacrylic acid
a multifunctional (PAA)
matrix thathydrogel
includesforreduced
glucose graphene
detection that and
integrates a multifunctional matrix that includes reduced graphene
lutetium phthalocyanine was reported. This biosensor provided high sensitivity for the detection and lutetium phthalocyanine wasof
reported. This biosensor provided high sensitivity for the detection
glucose with a low detection limit [26]. However, better control of the physical and chemical of glucose with a low detection
limit [26]. However,
properties is still neededbetter forcontrol of the physical
the fabrication of carbon andnanomaterial-based
chemical propertiesbiosensors,
is still needed suchfor as the
the
fabrication
miniaturization of the sensor, in vivo stability, separation processes for different type of CNTs,vivo
of carbon nanomaterial-based biosensors, such as the miniaturization of the sensor, in etc.
stability, separation processes
Other non-enzymatic sensors, for suchdifferent
as metallic typeoxide
of CNTs,
and etc. Other non-enzymatic
strontium palladium perovskite sensors, alsosuchshowas
metallic oxide and strontium palladium perovskite also show enhanced
enhanced sensitivity and improved detection limits, and thus avoid the need for expensive enzymes sensitivity and improved
detection
in the systemlimits, and thus avoid the need for expensive enzymes in the system [27,28].
[27,28].
Microgels,
Microgels,glucose-responsive
glucose-responsive polymer
polymer gels,gels,
havehave
several advantages
several as glucose
advantages sensing systems
as glucose sensing
due to their porous structures for carrying drugs, semi-solid properties
systems due to their porous structures for carrying drugs, semi-solid properties for easy engineering for easy engineering into
various shapes
into various (beads,
shapes films,films,
(beads, fibers, etc.),etc.),
fibers, and potential
and potentialbiocompatibility
biocompatibility for implantation.
for implantation. Since the
Since
first preparation of boronic acid-functionalized glucose sensitive
the first preparation of boronic acid-functionalized glucose sensitive hydrogels [29], many polymerhydrogels [29], many polymer
gel-based
gel-based glucose
glucosesensors
sensorshave havebeenbeen developed
developed based
based on on
the the
measurements
measurements of swelling
of swelling pressure [30],
pressure
fluorescence intensity
[30], fluorescence [31], changes
intensity in reflection
[31], changes holograms
in reflection [32,33] and
holograms diffraction
[32,33] of photonic
and diffraction colloidal
of photonic
crystal
colloidal crystal arrays [34]. Li et al. recently developed a series of inorganic-polymer optical
arrays [34]. Li et al. recently developed a series of inorganic-polymer hybrid microgels for hybrid
glucose sensing, based on the immobilization of fluorescent quantum
microgels for optical glucose sensing, based on the immobilization of fluorescent quantum dots dots (QDs) or metal nanoparticles
(NPs)
(QDs)intoor phenylboronic
metal nanoparticles acid (PBA)-functionalized
(NPs) into phenylboronic microgelsacid[35].(PBA)-functionalized
Zhang et al. constructed an advanced
microgels [35].
glucose-sensitive
Zhang et al. constructed an advanced glucose-sensitive platform on the basis beauty
platform on the basis of G1.0 PAMAM-functionalized microgels [36]. The of G1.0 of
this smart microgel, whichmicrogels
PAMAM-functionalized differentiates
[36]. itThefrom other of
beauty examples
this smartof traditional
microgel, glucose sensors, is thatit
which differentiates
itfrom
provides minimally invasive implantation and a fluorescent
other examples of traditional glucose sensors, is that it provides minimally signal by transdermal transmission
invasive
without
implantation and a fluorescent signal by transdermal transmission without any external of
any external links or electric power sources for CGM. A schematic illustration G1.0
links or
PAMAM-functionalized microgels that can recognize glucose and emit
electric power sources for CGM. A schematic illustration of G1.0 PAMAM-functionalized microgels blue fluorescence after injection,
is depicted
that in Figureglucose
can recognize 1. and emit blue fluorescence after injection, is depicted in Figure 1.

Figure 1.1. The

Figure The schematic
schematic illustration
illustration of
of G1.0
G1.0 PAMAM-functionalized
PAMAM-functionalized microgels
microgels that
that can
can recognize
recognize
glucose and emit blue fluorescence after injection. Reprinted with permission from [36].
glucose and emit blue fluorescence after injection. Reprinted with permission from [36].

Semiconductor quantum dots (QDs), as photoluminescent nanomaterials, have been intensively

Semiconductor quantum dots (QDs), as photoluminescent nanomaterials, have been intensively
studied in sensing and cell imaging due to their excellent performance and size-tunable optical
studied in sensing and cell imaging due to their excellent performance and size-tunable optical
properties [37]. Glucose could be detected by indirect measurements using photoelectrochemical QD
properties [37]. Glucose could be detected by indirect measurements using photoelectrochemical
sensors if combined with suitable enzymes [38,39]. Tanne et al. reported the indirect sensitive
detection of glucose by creating a signal chain from glucose via glucose oxidase and molecular
oxygen via CdSe/ZnS QDs toward the electrode [40], as shown in Figure 2. On the basis of the
Sensors 2017, 17, 182 5 of 19

QD sensors if combined with suitable enzymes [38,39]. Tanne et al. reported the indirect sensitive
Sensors 2016, 16, 182 5 of 18
detection of glucose by creating a signal chain from glucose via glucose oxidase and molecular oxygen
via CdSe/ZnS
influence QDs
that the towardconcentration
oxygen the electrode has[40],on
as the
shown in Figure 2.the
photocurrent, Onenzymatic
the basis of the influence
activity that
of glucose
the oxygen concentration has on the photocurrent, the enzymatic activity
oxidase catalyzing the oxidation of glucose by the reduction of O2 was evaluated. During of glucose oxidase catalyzing
the oxidation the
illumination, of glucose by the reduction
photocurrent was reducedof O2aswas evaluated.
a result of theDuring
oxygen illumination,
consumption. the photocurrent
The sensing
was reduced
properties as atype
of this resultofof the oxygen
electrode wereconsumption. The sensing
strongly influenced by theproperties
amount of ofenzyme
this typeonoftop
electrode
of the
QD layer, which was found to be easily adjustable using the layer-by-layer technique. to
were strongly influenced by the amount of enzyme on top of the QD layer, which was found Thebe
easily adjustableglucose
aforementioned using the layer-by-layer
sensors are based technique. The aforementioned
on signal chains of glucose-glucose glucose sensors are based
oxidase-oxygen-QDs.
on signal chains of glucose-glucose oxidase-oxygen-QDs. However, the
However, the enzyme can also be coupled to an electrode by means of a shuttle molecule, enzyme can also be coupled
which
to an electrode by means of a shuttle molecule, which facilitates a mediated
facilitates a mediated electron transfer from the biocatalyst. Zheng et al. constructed electron transfer from a
the biocatalyst. Zheng et al. constructed a photoelectrochemical electrode
photoelectrochemical electrode by alternately depositing water-soluble CdSe-CdS QDs and a by alternately depositing
water-soluble
mixture CdSe-CdS
of [Co(phen) QDs and a mixture of [Co(phen)3 ]2+/3+
3]2+/3+ and poly(ethyleneimine) on a TiO2 and poly(ethyleneimine)
electrode on a TiO
[41]. In this setup, the2
electrode [41]. In this setup, the electrode was able to transfer charge carriers
electrode was able to transfer charge carriers from the reduced enzyme, so that the obtained from the reduced enzyme,
so that the obtained
photocurrent depended photocurrent depended on
on the concentration the concentration of glucose.
of glucose.

Figure
Figure 2.
2. Illustration
Illustrationof
of the
the electron
electron transfer
transfer steps
steps after
after illumination
illumination of
of the
the QD
QD electrode.
electrode. Reprinted
Reprinted
with permission from [40].
with permission from [40].

Nowadays, label free and real time optical detection methods are the most promising
Nowadays, label free and real time optical detection methods are the most promising techniques in
techniques in glucose monitoring. Complementary metal oxide semiconductor (CMOS) image
glucose monitoring. Complementary metal oxide semiconductor (CMOS) image sensore are becoming
sensore are becoming powerful tools for biosensing applications with the advantages of high signal
powerful tools for biosensing applications with the advantages of high signal to noise ratio, low power
to noise ratio, low power consumption, miniaturization and availability on smartphones and even
consumption, miniaturization and availability on smartphones and even easier to incorporate into
easier to incorporate into compact medical diagnostic devices. In addition, CMOS image sensors can
compact medical diagnostic devices. In addition, CMOS image sensors can employ various glucose
employ various glucose monitoring methods, including continuous glucose monitoring methods
monitoring methods, including continuous glucose monitoring methods [42,43], enzymatic glucose
[42,43], enzymatic glucose monitoring methods [44,45], and thermal-based glucose monitoring [46],
monitoring methods [44,45], and thermal-based glucose monitoring [46], to operate at the in vitro and
to operate at the in vitro and in vivo level.
in vivo level.
Kim et al. exploited a CMOS image sensor for detecting glucose levels by simple photon count
variation with high sensitivity [44], as shown in Figure 3. Various concentrations of glucose (100 mg/dL
to 1000 mg/dL) were added onto a simple polydimethylsiloxane (PDMS) chip and the oxidation of
glucose was catalyzed with the aid of an enzymatic reaction. Oxidized glucose produces a brown
color with the help of a chromogen during the enzymatic reaction and the color density varies with
the glucose concentration. Photons pass through the PDMS chip with varying color density and hit
the sensor surface. Photon count was recognized by the CMOS image sensor depending on the color
density with respect to the glucose concentration and it was converted into digital form. By correlating
the obtained digital results with glucose concentration it is possible to measure a wide range of BG
levels with great linearity based on the CMOS image sensor and therefore this technique could promote
a convenient point-of-care diagnosis method.

Figure 3. Schematic of PDMS chip utilization for monitoring of glucose solutions by a CMOS image
sensor. Reprinted with permission from [44].
sensore are becoming powerful tools for biosensing applications with the advantages of high signal
to noise ratio, low power consumption, miniaturization and availability on smartphones and even
easier to incorporate into compact medical diagnostic devices. In addition, CMOS image sensors can
employ various glucose monitoring methods, including continuous glucose monitoring methods
[42,43],2017,
Sensors enzymatic
17, 182 glucose monitoring methods [44,45], and thermal-based glucose monitoring6[46],
of 19
to operate at the in vitro and in vivo level.

Figure 3. Schematic
Figure 3. Schematic of
of PDMS
PDMS chip
chip utilization
utilization for
for monitoring
monitoring of glucose solutions
of glucose solutions by
by aa CMOS
CMOS image
image
sensor. Reprinted with permission from [44].
sensor. Reprinted with permission from [44].

2.2. Novel Biocompatible Coatings for Sensors

The major challenge in the design of tissue-contacting materials for both bioaffinity and
medical implantation sensors is to endure protein adsorption between surfaces and interfaces.
Most minimally-invasive biosensors are surrounded by protective coatings that can strongly interact
with proteins, thereby minimizing tissue reactions induced by device implantation [47]. To achieve
a protein-resistant surface, the structure design of the anti-fouling material should be hydrophilic,
electrically neutral, and contain hydrogen bond acceptors instead of donors. These requirements
guide the design of anti-fouling formulations for sensing applications with novel structures that have
non-specific protein resistance [48]. A considerable number of effective candidates have been identified,
including organic/inorganic composites and biofunctional polymer architectures. Sol-gel derived
silicates have been demonstrated to be highly compatible with enzymes [49]. Silica-based hybrid
materials exhibit a fair biocompatibility both in vitro and in vivo, and are shown to be non-toxic and
biocompatible coatings for glucose oxidase-based sensors. Stable glucose responses were obtained for
the silica-polymer coated sensors both in buffered solutions containing bovine serum albumin and in
serum [50,51]. Unfortunately, the anti-fouling properties of such systems were not discussed.
Various macromolecules, including natural, semi-synthetic, and synthetic materials are currently
utilized in the fabrication of implantable CGM sensor coatings (cf. Table 1). Naturally occurring materials
such as alginate, chitin, and chitosan and their derivatives are widely investigated for biosensors in
the form of sheets, membranes and coatings, which offer the advantage of being comparable to
the natural precursors and the biological environment is prepared to recognize and deal with their
metabolites [52,53]. On the other hand, modified synthetic polymers with tailored structures and
coating properties are often superior to traditional macromolecules, especially for their reduced
immunogenicity. Commonly used biocomparable and biodegradable polymers includes poly(lactic
co-glycolic acid) (PLGA), poly(ethylene glycol) (PEG), poly(hydroxyethyl methacrylate) (PHEMA),
and poly(vinyl alcohol) (PVA), etc. [54]. Among the abovementioned polymers, HEMA-based materials
constitute the first generation of efficient protein-resistant materials while PEG-derived materials
represent the second generation [55]. Currently, they are extensively utilized in various areas and
can be used to modify mini-invasive devices by the coating method. They are also very popular as
hydrogel matrices, either alone or combined with other polymers [56].
Sensors 2017, 17, 182 7 of 19

Table 1. Biochemical aspects of commonly used polymers.

Polymer Characters Formation Applications

Natural
Alginate Immobilization of glucose oxidase Hydrogel and membrane Drug delivery
Collagen Extracellular matrix component Hydrogel, membrane and sponge Scaffolds
Semisynthetic
Chitin,
Hydrogel, membrane and fiber Anti-microbial and drug delivery
Chitosan
Synthetic
PLGA Negligible protein adsorption Micelle and hydrogel Coating, drug delivery and scaffolds
PHEMA Negligible protein adsorption Hydrogel Coating
PVA Geltaion and mechanical properties Hydrogel, membrane and sponge Coating and drug delivery
PEG Negligible protein adsorption Hydrogel and membrane Coating and drug delivery
PEGMA Negligible protein adsorption Hydrogel Coating

Hydrogels are hydrophilic, water-insoluble three-dimensional polymeric networks represented as

semi-open structures comprising entangled chains, which adsorb and store large amounts of water and
are highly permeable to small molecules. The use of hydrogel coatings allows the diffusion of glucose
through the swollen hydrogel layer. The degree of glucose diffusion could be readily modulated by
physically/chemically control of the cross-linking density of the hydrogel, which consequently affects
the water content and mechanical strength of polymer network. Both in vitro and in vivo animal
tests have demonstrated better performance with biocompatible polymer hydrogel outer layers for
a mini-invasive needle-type glucose sensor.
Despite oxidation problems that could alter the anti-fouling properties during long-term usage,
PEG-derived anti-fouling matrices remain popular and have been widely investigated [57]. Similar to
PEG, poly(ethylene glycol) methyl ether methacrylate (PEGMA) and PEGMA-analogous hydrogels
were also investigated for the design of the next generation of anti-biofouling materials [58]. The results
revealed that the molecular weight of the PEGMA homopolymer directly affected the overall
hydrophilicity of the gel, which could be attributed to both the intermediate hydrophilic chain length
and intermediate crosslinking degree. Furthermore, the results of micro-biofouling by bacteria and
blood cells demonstrated that PEGMA hydrogels offered the perfect non-fouling ability, showing
application potential for the coating of devices in contact with blood and the in vivo controlled release
of drugs from nonfouling hydrogels.

2.3. Drug Deliver and Its System

Drug delivery is a potential field of application for CGM devices [59]. In recent years, architectures
for sustained and controlled delivery of drugs has been extensively studied due to several problems
in common drug delivery systems, such as limited stability, inopportune leakage and poor solubility,
especially for CGM systems. Modern closed-loop BG controlling systems have been developed by
various organizations around the world, by merging glucose-sensing devices and insulin-delivery
carriers for optimized dosage of insulin according to detected glycemic levels (Figure 4). Such BG
management systems utilize a ‘Sense and Act’ feedback-loop design to realize appropriate corrective
action by timely and optimal dosing. The pump dispenses the right amount of insulin as the sensor
relays the information. The development of these responsive drug delivery systems is anticipated to
dramatically change diabetes management and patient monitoring. It is expected that the release of
different drugs could be controlled by a microprocessor in response to the signal from a biosensor via
different release kinetics.
Based on the biocoating, intelligent drug releasing functionality was also investigated to make
the system more integrated, and ensure the long-term usage of the device [60]. For instance, a PLGA-
copolymer, PVA and PEG combined system which has been shown to achieve a continuous release of
dexamethasone for up to one month that decreases extravasation of leukocytes, and inhibits production
of proteolytic enzymes and chemoattractive factors was already discussed [61,62]. A masitinib-releasing
glucose oxidase on the biosensor. The result showed that the sensitivity of the as-prepared spiral
sensor was 20 nA/(mmol/L)~30 nA/(mmol/L), which provided a linear detection of physiological
glucose concentrations between 2 and 30 mmol/L. The sensors also showed good repeatability,
selectivity, and stability, which could be tuned by adjusting the amount of the polyurethane 8(PU)
Sensors 2017, 17, 182 of 19
semi-permeable membrane coating [64]. More recently, the PU coating was further modified by a
PVA-PEG hydrogel that enabled zero-ordered release of dexamethasone, and the release duration
PLGA carrier delivers
was controlled locally from
as a function soluble
of film PEG-coated
gelation density. CGM sensors were
The cumulative developed
released to reduce
amount couldthe
be
host-implant response. The releasing action could be attributed to the CGM sensor output
adjusted by changes in the molecular weight of the PEG. As a matter of fact, PEG derivatives were fluctuations
over time,towhich
revealed was monitored
be effective for 21
candidates fordays. Thecoatings
sensor results showed
as well relatively improved
as controlled performance
release of supposed in
terms
drugs of sensing ability in the drug-delivery implant group compared to the control group [63].
[58,65].

Figure 4.4.Illustration
Figure Illustrationofof closed-loop
closed-loop glycemic
glycemic management
management system
system utilizing
utilizing the and
the ‘Sense ‘Sense
Act’and Act’
method
method for optimized insulin
for optimized insulin delivery. delivery.

3. Non-Invasive Technology
Recently, a novel implantable electrochemical sensor was fabricated by utilizing a spiral Pt-Ir
alloyNon-invasive
electrode to increase
techniquesthe have
activereceived
area of the workingresearch
significant electrode as well
interest dueas to
enhance the loading
the highly of
sensitivity
glucose
and betteroxidase
patienton compliance,
the biosensor.contrary
The result
to showed
invasivethat
ones.theTypical
sensitivity of the as-prepared
non-invasive spiral
biosensors sensor
based on
was 20 nA/(mmol/L)~30
different approaches include nA/(mmol/L),
iontophoretic which provided
extraction a linearthrough
of glucose detection of skin,
the physiological glucose
surface plasmon
concentrations
resonance, Raman between 2 and 30 mmol/L.
spectroscopy in aqueousThe sensors
humor, also showed
visible good repeatability,
or near-infrared (NIR) selectivity, and
spectroscopy,
stability, which
polarimetry, could be tuned
photo-acoustic by adjusting
probes, the amount of
and fluorescence the polyurethane
methods. Although(PU) semi-permeable
results from these
membrane
innovative coating
devices [64]. More
require recently,
frequent the PU coating
calibration againstwas further
direct modified
BG data, theyby a PVA-PEG
might hydrogel
be an alternative
that
choiceenabled zero-ordered
for future continuousrelease of dexamethasone,
glucose measurement. and the release the
Consequently, duration
challengewas controlled
of preparing as
aaccurate
functionlevel
of film gelation
sensors to density.
biomonitor The conveniently
cumulative released amount could
and painlessly be adjustedremains
BG information by changes in
to be
the molecular weight of the PEG. As a matter of fact, PEG derivatives were revealed to be effective
solved.
candidates for sensor coatings as well as controlled release of supposed drugs [58,65].
3.1. Tear Sensing Designs
3. Non-Invasive Technology
Non-invasive, contact lenses are one of the most appealing ways to achieve constant contact
with Non-invasive
tear fluid andtechniques have received
monitor glucose in tears.significant research
Such a method interest
could duefor
be used to normal
the highly sensitivity
conditions as
and
well better
as forpatient compliance,
clinical contrary
trials. During the to invasive
course ones. Typical
of treatment, thenon-invasive
glucose could biosensors based on
be continuously
different
monitored. approaches include
Contact lenses iontophoretic
have extraction
to meet a certain of glucose
number through
of criteria, andthe skin, surface
should be madeplasmon
of soft,
resonance, Raman spectroscopy in aqueous humor, visible
comfortable materials with constant mechanical properties such as hydrogels oror near-infrared (NIR) spectroscopy,
plasticized
polarimetry,
polymers that photo-acoustic
could withstand probes,
theand
eyefluorescence
movements methods.
and blinking.Although resultsthe
Moreover, from thesealso
lenses innovative
should
devices require frequent calibration against direct BG data, they might be an alternative
be non-toxic and to allow for a certain amount of oxygen and tear fluid to pass through to the eye. choice for
future continuous glucose
Some interactions betweenmeasurement.
contact lenses Consequently,
and the tear filmthe challenge of preparing
were discussed. accurateoflevel
The presence the
sensors to biomonitor conveniently and painlessly BG information remains to
contact lens immediately provides the potential for both biophysical and biochemical phenomena. be solved.
The biophysical phenomena resulting from lens-tear interactions are often directly observable as the
3.1. Tear Sensing Designs
increased osmotic pressure caused by enhanced evaporation of the tear film. The biochemical
Non-invasive, contact lenses are one of the most appealing ways to achieve constant contact with
tear fluid and monitor glucose in tears. Such a method could be used for normal conditions as well
as for clinical trials. During the course of treatment, the glucose could be continuously monitored.
Contact lenses have to meet a certain number of criteria, and should be made of soft, comfortable
materials with constant mechanical properties such as hydrogels or plasticized polymers that could
Sensors 2017, 17, 182 9 of 19

withstand the eye movements and blinking. Moreover, the lenses also should be non-toxic and to allow
for a certain amount of oxygen and tear fluid to pass through to the eye. Some interactions between
contact lenses and the tear film were discussed. The presence of the contact lens immediately provides
the potential
Sensors 2016, 16, for
182 both biophysical and biochemical phenomena. The biophysical phenomena resulting 9 of 18
from lens-tear interactions are often directly observable as the increased osmotic pressure caused by
outcomes
enhanced revealed
evaporation the of
difference in electrolyte
the tear film. concentration
The biochemical outcomesbetween the the
revealed anterior and posterior
difference tear
in electrolyte
films [66,67]. between the anterior and posterior tear films [66,67].
concentration
Several
Several types
types ofof sensors
sensors have
have emerged
emerged in in recent
recent years
years based
based onon different
different monitoring
monitoring methods,
methods,
including
including electrochemical
electrochemical and and spectral determination.
determination. Electrochemical
Electrochemical sensors
sensors have
have become
become more
complicated and miniaturized at the same time. A A protocol
protocol consisting of a soft soft polymer
polymer lens
lens covered
covered
by an enzyme-based
enzyme-based sensor sensor waswas reported,
reported, using
using aa tiny
tiny metal
metal electrode
electrode laid
laid out
out of
of the lens to detect
electrochemical signalsinin
electrochemical signals response
response to glucose
to glucose [68]. [68]. Another
Another designdesign
includedincluded integration
integration of an
of an electrical
electrical circuit, probe,
circuit, a sensing a sensing probe, a body-wired
a body-wired communication communication
transmitter, atransmitter,
power supply, a power
and a supply, and a
PDMS contact
PDMS
lens by contact
using MEMSlens by using MEMS
techniques. During techniques. During
the monitoring, thethe monitoring,
glucose solutionthewasglucose solution was
orally administrated
orally administrated
to rabbits and the BG to rabbits
level and the BG
was measured level
using was measured
a glucose monitoringusing
kit a(Figure
glucose5). monitoring
This biosensor kit
(Figure
showed 5). This
rapid biosensor
responses to showed rapid
glucose and anresponses
appropriate to calibration
glucose and an appropriate
range (0.03–5.0 mM), calibration range
which covered
(0.03–5.0
the reportedmM), which
tear covered
glucose levelsthe reported tear glucose levels [69].
[69].

Figure
Figure 5.
5. Measurement
Measurement method
method of
of tear
tear glucose
glucose concentration
concentration with
with aa contact
contact lens
lens biosensor.
biosensor. BG
BG levels
levels
were simultaneously
simultaneouslymeasured
measuredbybya commercial
a commercialBGBG
monitoring kit. Reprinted
monitoring with with
kit. Reprinted permission from
permission
[69].
from [69].

Spectral sensors for CGM, on the other hand, were simplified. Contact glucose-sensing lenses
Spectral sensors for CGM, on the other hand, were simplified. Contact glucose-sensing lenses based
based on hydrogels that contain phenylboronic acid (PBA) and photonic crystals that function
on hydrogels that contain phenylboronic acid (PBA) and photonic crystals that function without requiring
without requiring power were proposed [70–72]. Such soft hydrogel lenses embedded an array of
power were proposed [70–72]. Such soft hydrogel lenses embedded an array of mono-disperse particles,
mono-disperse particles, which could diffract electromagnetic waves according to Bragg’s law. It is
which could diffract electromagnetic waves according to Bragg’s law. It is well known that PBA-modified
well known that PBA-modified hydrogels are able to bind cis-diols in glucose, leading to volumetric
hydrogels are able to bind cis-diols in glucose, leading to volumetric changes and forcing the lens to
changes and forcing the lens to expel water, altering the array spacing, thus changing the perceived
expel water, altering the array spacing, thus changing the perceived color of the lens. Moreover, the color
color of the lens. Moreover, the color changes caused by the reaction are quick and reversible. Other
changes caused by the reaction are quick and reversible. Other photonic crystal embedded lenses were
photonic crystal embedded lenses were developed to improve the sensitivity or use different
developed to improve the sensitivity or use different detection spectrometers [73,74]. Recently, a plastic
detection spectrometers [73,74]. Recently, a plastic contact lens outfitted with a physically gelated
contact lens outfitted with a physically gelated photonic crystal hydrogel was synthesized for CGM
photonic crystal hydrogel was synthesized for CGM in our group. The hydrogel was physically
in our group. The hydrogel was physically crosslinked from PVA-PEG system, which could offer
crosslinked from PVA-PEG system, which could offer an anti-fouling layer for the penetration of
an anti-fouling layer for the penetration of glucose. This hydrogel was subsequently modified with
glucose. This hydrogel was subsequently modified with 4-boronobenzaldehyde, inproving the
sensitivity for glucose, and a prototype of such a lens is shown in Figure 6a. The sensing behavior of
the lens was monitored by a reflection photospectrometer, and the results showed that the diffracted
wavelength was relatively shifted with the increasing glucose concentration in artificial tear solution
(Figure 6b). Despite the exciting design, it remains difficult to measure glucose concentrations in
tears using contact lenses. Because of the very low amount of glucose present in healthy control
Sensors 2017, 17, 182 10 of 19

4-boronobenzaldehyde, inproving the sensitivity for glucose, and a prototype of such a lens is shown in
Figure 6a. The sensing behavior of the lens was monitored by a reflection photospectrometer, and the
results showed that the diffracted wavelength was relatively shifted with the increasing glucose
concentration
Sensors 2016, 16, in
182artificial tear solution (Figure 6b). Despite the exciting design, it remains difficult
10 of 18
to measure glucose concentrations in tears using contact lenses. Because of the very low amount of
glucose
subjectspresent
(3.59 mM)in healthy
and forcontrol subjects
diabetic (3.59
subjects mM)
(4.69 mM),andthe
fortear
diabetic subjects
glucose levels (4.69 mM), appear
measured the tearto
glucose
vary withlevels
themeasured
volume ofappear to varytear
the aqueous with the volume
fraction of the aqueous tear fraction collected.
collected.

(a) (b)
Figure 6. (a) Diagram and photograph (insert) of a physical hydrogel photonic crystal sensing lens;
Figure 6. (a) Diagram and photograph (insert) of a physical hydrogel photonic crystal sensing lens;
(b)
(b) Diffraction
Diffraction wavelength
wavelength shifts
shifts withwith the variation
the variation of the glucose
of the glucose concentration
concentration intear
in artificial artificial tear
solution.
solution.
3.2. Salivary Biosensors
3.2. Salivary Biosensors
Many authors have found higher glucose salivary levels in diabetic patients than in
Many authors have found higher glucose salivary levels in diabetic patients than in
non-diabetics [75–78]. Such investigations mainly aimed at exploring whether diabetic control could be
non-diabetics [75–78]. Such investigations mainly aimed at exploring whether diabetic control could
monitored by a non-invasive salivary glucose measurement method. Saliva is a great diagnostic fluid
be monitored by a non-invasive salivary glucose measurement method. Saliva is a great diagnostic
providing an alternative to direct blood analysis via the permeation of blood constituents without any
fluid providing an alternative to direct blood analysis via the permeation of blood constituents
skin-piercing for blood sampling [79]. Actually, saliva glucose concentrations range approximately from
without any skin-piercing for blood sampling [79]. Actually, saliva glucose concentrations range
20 to 200 mmol/L in normal and diabetic individuals, and closely follow circadian BG fluctuations [80].
approximately from 20 to 200 mmol/L in normal and diabetic individuals, and closely follow
Saliva and BG levels correlate reasonably well in a sample of individuals [81–83], however, a much
circadian BG fluctuations [80]. Saliva and BG levels correlate reasonably well in a sample of
stronger correlation is observed within the same individual, enabling BG concentrations to be estimated
individuals [81–83], however, a much stronger correlation is observed within the same individual,
from saliva glucose measurements [84].
enabling BG concentrations to be estimated from saliva glucose measurements [84].
Various attempts have been made by researchers to develop glucose monitoring systems for
Various attempts have been made by researchers to develop glucose monitoring systems for
salivary measurements. Lambert et al. used conventional measurement through colorimetric assays [85].
salivary measurements. Lambert et al. used conventional measurement through colorimetric assays
Lipson et al. detected the saliva glucose level through NIR/Raman-based spectroscopic techniques [86],
[85]. Lipson et al. detected the saliva glucose level through NIR/Raman-based spectroscopic
and Yamaguchi et al. used Clarke-type electrode-based amperometric measurements, etc. [80]. Taking clues
techniques [86], and Yamaguchi et al. used Clarke-type electrode-based amperometric
from these developments, Soni et al. developed an optical biosensor for direct determination of salivary
measurements, etc. [80]. Taking clues from these developments, Soni et al. developed an optical
glucose by using glucose oxidase enzyme immobilized on filter paper strips (specific activity 1.4 U/strip)
biosensor for direct determination of salivary glucose by using glucose oxidase enzyme immobilized
and then reacting it with synthetic glucose samples in the presence of a co-immobilized color pH
on filter paper strips (specific activity 1.4 U/strip) and then reacting it with synthetic glucose samples
indicator [83]. The filter paper changed color based on the concentration of glucose in the reaction
in the presence of a co-immobilized color pH indicator [83]. The filter paper changed color based on
medium and hence, by scanning this color change (using RGB profiling) through an office scanner
the concentration of glucose in the reaction medium and hence, by scanning this color change (using
and using open source image processing software (GIMP), the concentration of glucose in the reaction
RGB profiling) through an office scanner and using open source image processing software (GIMP),
medium could be deduced.
the concentration of glucose in the reaction medium could be deduced.
Wearable sensors have recently attracted considerable interest owing to their promise for real-time
Wearable sensors have recently attracted considerable interest owing to their promise for
monitoring of the wearer’s health and fitness in a wide range of biomedical, sport and military
real-time monitoring of the wearer’s health and fitness in a wide range of biomedical, sport and
scenarios [87,88]. Recent efforts have led to wearable biosensors for detecting chemical biomarkers
military scenarios [87,88]. Recent efforts have led to wearable biosensors for detecting chemical
in human fluids that can be obtained non-invasively, e.g., tears, sweat or saliva [89–92]. Kim et al.
biomarkers in human fluids that can be obtained non-invasively, e.g., tears, sweat or saliva [89–92].
reported a mouth-guard biosensor for continuous monitoring of salivary lactate and other chemical
Kim et al. reported a mouth-guard biosensor for continuous monitoring of salivary lactate and other
components [79,93]. A mouth-guard glucose sensor produced using micro electromechanical systems
chemical components [79,93]. A mouth-guard glucose sensor produced using micro
electromechanical systems (MEMS) techniques would offer promise as a minimally-invasive,
painless, continuous, custom-fitted and wireless solution for self-monitoring of glucose.
Mitsubayashi et al. have developed a detachable “Cavitas sensor” used in the human oral cavity for
non-invasive monitoring of saliva glucose [94], as shown in Figure 7. The mouth-guard glucose
Sensors 2017, 17, 182 11 of 19

(MEMS) techniques would offer promise as a minimally-invasive, painless, continuous, custom-fitted

and wireless solution for self-monitoring of glucose. Mitsubayashi et al. have developed a detachable
“Cavitas sensor” used in the human oral cavity for non-invasive monitoring of saliva glucose [94],
as shown
Sensors 2016, in
16, Figure
182 7. The mouth-guard glucose sensor consisted of a platinum and silver/silver
11 of 18
chloride electrode, with glucose oxidase immobilized by entrapment with poly(MPC-co-EHMA)
(PMEH),
mouth-guard on a custom-fitted
support with monolithic
a wireless mouth-guard supportenabling
transmitter, thereby with a wireless transmitter,
telemetric thereby
measurement of
enabling telemetric measurement of saliva glucose.
saliva glucose.

Figure 7.
Figure 7. (A)
(A) Schematic
Schematic image
image ofof the
the glucose biosensor on
glucose biosensor on the
the polyethylene
polyethylene terephthalate
terephthalate glycolglycol
mouthguard support. Pt and Ag electrodes were formed on the PETG through
mouthguard support. Pt and Ag electrodes were formed on the PETG through a sputtering process. a sputtering process.
Each electrode
Each electrode sensor
sensor consisted
consisted ofof aa 0.20
0.20 mm
mm22 PtPt working
working electrode
electrode andand aa 4.0
4.0mm mm Ag/AgCl
2 2 Ag/AgCl
reference/counter electrode, both insulated with PDMS on a 0.5 mm thick PETG
reference/counter electrode, both insulated with PDMS on a 0.5 mm thick PETG layer. 30 units of layer. 30 units
GOD of
GOD were applied to the sensing region of the working electrode. In order to
were applied to the sensing region of the working electrode. In order to optimize enzyme entrapment, optimize enzyme
entrapment,
2.0 mL of 1.02.0 wt%mLPMEH
of 1.0 wt% PMEH
solution was solution
spread was
overspread over the
the sensing sensing
region region
to form thetoPMEH
form the PMEH
overcoat;
overcoat;
(B) (B) Schematic
Schematic image of the image of the mouth-guard
mouth-guard biosensor to
biosensor custom-fit custom-fit to the
the patient’s patient’sThe
dentition. dentition.
device
The device
consists of a consists
glucose of a glucose
sensor sensor transmitter
and wireless and wireless transmitter incorporating
incorporating a potentiostat fora potentiostat
stable glucose for
stable glucose measurement. The sensor was designed to fit the mandibular
measurement. The sensor was designed to fit the mandibular dentition from the first premolar updentition from the first
premolar
to upmolar.
the third to theThe
thirdwireless
molar. The wirelesswas
transmitter transmitter was neatly
neatly encased encased
in PETG. in PETG.with
Reprinted Reprinted with
permission
permission
from [94]. from [94].

3.3. Other
3.3. Other Methods
Methods
Recently continuous
Recently continuousglucose
glucosemonitoring
monitoring devices
devices havehave become
become available
available and could
and could provide
provide more
more detailed data on glucose excursions. In future applications such continuous
detailed data on glucose excursions. In future applications such continuous glucose sensors may glucose sensors
may become
become a critical
a critical component
component of closedof loop
closed loop delivery
insulin insulin delivery
systems systems and, must
and, as such, as such, must be
be selective,
selective,
rapid, rapid, predictable
predictable and for
and acceptable acceptable
continuous for patient
continuous patient
use. Many use. Many
potential potential
sensing sensing
modalities are
modalities are being pursued for this, including
being pursued for this, including optical techniques. optical techniques.
Optical sensors
Optical sensors use
use light
light of
of variable
variable frequencies
frequencies to to detect
detect glucose,
glucose, utilizing
utilizing different
different interaction
interaction
properties of
properties of light
light with
with glucose
glucose molecules
molecules in in aa concentration-dependent
concentration-dependent manner
manner [95].
[95]. Compared
Compared to to
conventional glucose sensors, optical sensors benefit from the absence of electromagnetic
conventional glucose sensors, optical sensors benefit from the absence of electromagnetic interference,
interference, simple design and handling as well as low cost. Miniaturization of optical sensors is
easily achievable without compromising performance and dramatically reduces repulsion reactions.
Evidently, optical sensing technology can provide an interesting alternative to more established
electrochemical sensors and will contribute to better flexibility concerning the design of CGM
systems, their cost, materials used, etc. [96]. Based on this optical transducing technology, numerous
Sensors 2017, 17, 182 12 of 19

simple design and handling as well as low cost. Miniaturization of optical sensors is easily achievable
without compromising performance and dramatically reduces repulsion reactions. Evidently, optical
sensing technology can provide an interesting alternative to more established electrochemical sensors
and will contribute to better flexibility concerning the design of CGM systems, their cost, materials
used, etc. [96]. Based on this optical transducing technology, numerous optics-based CGM systems
have been developed, such as fluorescence, infrared absorption spectroscopy, and Raman spectroscopy.
Fluorescence uses the principle of varying light emission from molecules in different states. It is
fast, reagentless and extremely sensitive. Many fluorescence-based glucose sensors are based upon the
affinity sensor principle where glucose and a fluorescein-labelled analogue bind competitively with
a receptor site specific for both ligands [97,98]. Other fluorescence techniques use enzyme- catalysed
reactions to change fluorescent status. Fluorescein may be bound to glucose oxidase, allowing energy
transfer between the flavin group of GOx and the fluorescein. The GOx enzyme may be further
utilized by measuring oxygen consumption or hydrogen peroxide production by fluorescence [99,100].
GOx can also be used as glucose-binding protein, utilizing its intrinsic fluorescent properties and may
be used without its flavin group with a fluorescent label [101].
Near infrared (NIR) light (wavelength 0.7–1.4 µm) provides an optical window in which 90%–95%
of light passes through the stratum corneum and epidermis into the subcutaneous space independent
of skin pigmentation. It is already successfully used to non-invasively monitor the concentration of
oxygenated and deoxygenated haemoglobin in the preterm infant brain [102]. BG has been measured
across the oral mucosa using NIR [103]. The oral mucosa is an obvious candidate for sensing, as it
is well vascularized and may be trans-illuminated. However, any oral measurement includes saliva,
which may contain a different glucose concentration, and residual food may contain interferents.
NIR spectroscopy has been measured across the tongue [104]. The standard error of prediction was
3.5 mmol/L, with significant variability caused by variability in tissue fat. NIR therefore ideally
requires consistent tissue fat in addition to a path length in the order of 5 mm [105]. In order to solve
some of the problems posed by transdermal optical techniques, NIR has been proposed in combination
with techniques for sampling ISF, including sonophoresis [106].
Raman spectroscopy assesses scattering of single wavelength light. This is dependent on rotational
or vibrational energy states within a molecule and highly specific absorption bands are seen with
Raman spectroscopy which can be used to identify and quantify molecules. It has the benefit of
reduced interference from water compared with MIR or NIR spectroscopy. However, the Raman
signal is weaker than that of other technologies, requiring powerful detectors for physiological
concentrations of glucose. The Raman signal is also susceptible to turbidity, haematocrit, skin thickness
and melanin. Sufficient sensitivity can be achieved by surface-enhanced Raman spectroscopy (SERS).
The electromagnetic mechanism of SERS is that when an electromagnetic wave interacts with a metal
surface, the fields at the surface are different than those observed in the far field. If the surface is
rough, the wave may excite localized surface plasmons on the surface, resulting in amplification of
the electromagnetic fields near the surface. If one assumes that there is enhancement of the intensity
of the incident and scattered fields (albeit at different wavelengths), then the possibility of a large
enhancement of Raman scattering intensity arises [107]. And now some reports utilized the SERS
signal probes served as the molecular recognition agent in response to the concentration of glucose.
Torul et al. fabricated a SERS substrate by modifying two component self-assembled monolayers
(SAMS) on the surface of gold nanorod particles [108]. The variation of the SERS signal of SAMS
showed the response to the concentration of glucose, and a low detection limit of 0.5 mM was obtained.
Kong et al. proposed a novel glucose binding mechanism by using phenylboronic acid as the receptor
for saccharide and forming a glucose-alkyne-boronic acid complex on SERS substrate, which exhibited
a new Raman peak at 1996 cm−1 [109]. Thus, the novel technology offered a high sensitivity for SERS
glucose sensing.
Sensors 2017, 17, 182 13 of 19

4. Future Perspectives
The worldwide increase in the number of diabetic patients has encouraged scientists to put great
efforts into the field of continuous glucose monitoring (CGM) devices, which would dramatically alter
the treatment of diabetes as well as the life quality of diabetic patients. So far, there are only a few
commercial CGM devices, developed by Medtronic (Minneapolis, MN, USA) and Dexcom, which have
been approved by the FDA and their current lifetimes are less than one week. No commercial CGM
devices are available for long-term monitoring, however, it is extremely important for T1D, which
accounts for around 10% of diabetic patients. The existing studies of long-term mini-invasive biosensors
remain challenging, mainly due to the foreign body responses such as biofouling, fibrous capsule
and inflammation. To overcome foreign body responses and improve the life-time of implantable
biosensors, we believe the following issues should be addressed: (1) improving the loading amount
and stability of enzymes. The coil type electrode design is one way to load more enzyme to reduce
the effects from accumulation of H2 O2 , glucose oxidase molecular cloning or encapsulation in silica
sol-gel may be another way to reduce the degradation of cross-linked enzymes [110]; (2) a number
of interferences (e.g., acetaminophen, ascorbic acid, and uric acid) can affect sensor response as they
are electroactive during the oxidation of hydrogen peroxide, although semi-permeable membranes
with designed porosity, like polyurethane, are able to alleviate this effect, At the meantime, decreasing
the work potential through electrode design is another route to reduce interference; (3) to reduce
foreign body responses once the biosensors are implanted, the synthesis of anti-fouling films is
a common strategy to improve glucose sensor functionality against the initial adhesion of proteins.
The introducing of biocompatible hydrogel coatings or collagen encapsulation for sensing devices are
other key approaches to improve the lifetime. The use of small anti-inflammatory and/or antigenic
molecules, such as dexamethasone, to influence tissue integration has been intensively studied. Here,
the drug delivery system with biocompatible and controllable particles, like poly(lactic-co-glycolic acid)
is pursued to limit inflammation and take advantage of the wound healing properties of this material.
Despite the fact that the results from non-invasive devices require frequent calibration against
invasive ones, the innovative design might be an alternative choice for glucose measurements than
traditional implanted electrochemical biosensors. Two possible techniques have the most chance
for non-invasive glucose monitoring, one is tear glucose sensors and another is salivary biosensors.
Detecting glucose in tear fluid has been established for over twenty years, and the major challenge
for traditional contact lens biosensors is that the amount tear fluid is not high enough to cover the
electrode to construct an electrochemical cell, and this thus leads to less accuracy in this type of
sensor. Compared with electrochemical sensors, a plastic contact lens outfitted with a physically
gelated photonic crystal hydrogel maybe a possible route for CGM in the future. A salivary biosensor
incorporating Pt and Ag/AgCl electrodes on a mouth-guard support as dental material with an enzyme
membrane was developed and tested by Mitsubayashi’s group [94], and this invisible mouth-guard
biosensor integrated with a low energy wireless module could be an ideal candidate for future real-time
monitoring of salivary glucose levels.
Non-enzymatic sensor show enhanced sensitivity and detection limits, while avoiding the
expensive and fragile enzymes in the system, and thus it is also a promising choice for the next
generation of CGM. Although there are large number of research work on this area, the clinic products
are not yet available in the market. The ultimate goal for treating diabetic patients is to construct an
artificial pancreas, combining CGM and insulin dosage system, the life time of CGM is the greatest
challenge, and are required for deeply and comprehensively studied in the future.

Acknowledgments: This work was supported by the National Natural Science Foundation of China (61471233,
21504051), the Program for Professor of Special Appointment (Eastern Scholar) at SIHL, the Sailing Project and
Basic Research Program from Science and Technology Commission of Shanghai Municipality (14YF1410600,
14JC1406402), Shuguang and ChenGuang project supported by Shanghai Municipal Education Commission
and Shanghai Education Development Foundation (14SG52, 13CG62), the key subject of Shanghai Polytechnic
University (Material Science and Engineering, XXKZD1601).
Sensors 2017, 17, 182 14 of 19

Conflicts of Interest: The authors declare no conflict of interest.

References
1. World Health Organization (WHO). Global Report on Diabetes 2016; WHO: Geneva, Switzerland, 2016.
2. Centers for Disease Control and Prevention. National Diabetes Fact Sheet: National Estimates and General
Information on Diabetes and Prediabetes in the United States; Centers for Disease Control and Prevention: Atlanta,
GA, USA, 2011.
3. Wild, S.; Roglic, G.; Green, A.; Sicree, R.; King, H. Global prevalence of diabetes: Estimates for the year 2000
and projections for 2030. Diabetes Care 2004, 27, 1047–1053. [CrossRef] [PubMed]
4. Chase, H.P.; Beck, R.W.; Xing, D.; Tamborlane, W.V.; Coffey, J.; Fox, L.A.; Ives, B.; Keady, J.; Kollman, C.;
Laffel, L.; et al. Continuous glucose monitoring in youth with type 1 diabetes: 12-month follow-up of the
Juvenile Diabetes Research Foundation continuous glucose monitoring randomized trial. Diabetes Technol. Ther.
2010, 12, 507–515. [CrossRef] [PubMed]
5. Vigersky, R.A. The benefits, limitations, and cost-effectiveness of advanced technologies in the management
of patients with diabetes mellitus. J. Diabetes Sci. Technol. 2015, 9, 320–330. [CrossRef] [PubMed]
6. Ehrhardt, N.M.; Chellappa, M.; Walker, M.S.; Fonda, S.J.; Vigersky, R.A. The effect of real-time continuous
glucose monitoring on glycemic control in patients with type 2 diabetes mellitus. J. Diabetes Sci. Technol.
2011, 5, 668–675. [CrossRef] [PubMed]
7. Clark, L.C.; Lyon, C. Electrode systems for continuous monitoring in cardiovascular surgery. Ann. N. Y.
Acad. Sci. 1962, 102, 29–45. [CrossRef] [PubMed]
8. Dovč, K.; Bratina, N.; Battelino, T. A new horizon for glucose monitoring. Horm. Res. Paediatr. 2015, 83,
149–156. [CrossRef] [PubMed]
9. Mauras, N.; Fox, L.; Englert, K.; Beck, R.W. Continuous glucose monitoring in type 1 diabetes. Endocrine
2013, 43, 41–50. [CrossRef] [PubMed]
10. McGarraugh, G.; Bergenstal, R. Detection of hypoglycemia with continuous interstitial and traditional blood
glucose monitoring using the FreeStyle navigator continuous glucose monitoring system. Diabetes Technol. Ther.
2009, 11, 145–150. [CrossRef] [PubMed]
11. Sparacino, G.; Facchinetti, A.; Cobelli, C. “Smart” continuous glucose monitoring sensors: Online signal
processing issues. Sensors 2010, 10, 6751–6772. [CrossRef] [PubMed]
12. Cobelli, C.; Renard, E.; Kovatchev, B. Artificial pancreas: Past, present, future. Diabetes 2011, 60, 2672–2682.
[CrossRef] [PubMed]
13. Langendam, M.; Luijf, Y.M.; Hooft, L.; DeVries, J.H.; Mudde, A.H.; Scholten, R.J.P.M. Continuous glucose
monitoring systems for type 1 diabetes mellitus. Cochrane Database Syst. Rev. 2012, 1, CD008101. [PubMed]
14. McQueen, R.B.; Ellis, S.L.; Campbell, J.D.; Nair, K.V.; Sullivan, P.W. Cost-effectiveness of continuous glucose
monitoring and intensive insulin therapy for type 1 diabetes. Cost Eff. Resour. Alloc. 2011, 9, 13. [CrossRef]
[PubMed]
15. Szypowska, A.; Ramotowska, A.; Dżygało, K.; Golicki, D. Beneficial effect of real-time continuous glucose
monitoring system on glycemic control in type 1 diabetic patients: Systematic review and meta-analysis of
randomized trials. Eur. J. Endocrinol. 2012, 166, 567–574. [CrossRef] [PubMed]
16. Zhu, Z.G.; Garcia-Gancedo, L.; Flewitt, A.J.; Xie, H.Q.; Moussy, F.; Milne, W.I. A Critical Review of Glucose
Biosensors Based on Carbon Nanomaterials: Carbon Nanotubes and Graphene. Sensors 2012, 12, 5996–6022.
[CrossRef] [PubMed]
17. Aggidis, A.G.A.; Newman, J.D.; Aggidis, G.A. Investigating pipeline and state of the art blood glucose
biosensors to formulate next steps. Biosens. Bioelectron. 2015, 74, 243–262. [CrossRef] [PubMed]
18. Bailey, T.; Bode, B.W.; Christiansen, M.P.; Klaff, L.J.; Alva, S. The performance and usability of a factory-
calibrated flash glucose monitoring system. Diabetes Technol. Ther. 2015, 17, 787–794. [CrossRef] [PubMed]
19. Wu, C.J.; Wu, S.Y.; Chen, P.C.; Lin, Y.S. An Innovative Smartphone-Based Otorhinoendoscope and Its Application
in Mobile Health and Teleotolaryngology. J. Med. Internet. Res. 2014, 16, e71. [CrossRef] [PubMed]
20. Zhu, Z.G.; Garcia-Gancedo, L.; Chen, C.; Zhu, X.R.; Xie, H.Q.; Flewitt, A.J.; Milne, W.I. Enzyme-free glucose
biosensor based on low density CNT forest grown directly on a Si/SiO2 substrate. Sens. Actuators B Chem.
2013, 178, 586–592. [CrossRef]
Sensors 2017, 17, 182 15 of 19

21. Wang, H.C.; Zhou, H.; Chen, B.Q.; Mendes, P.M.; Fossey, J.S.; James, T.D.; Long, Y.T. A bis-boronic acid
modified electrode for the sensitive and selective determination of glucose concentrations. Analyst 2013, 138,
7146–7151. [CrossRef] [PubMed]
22. Moon, B.; de Vries, M.G.; Cordeiro, C.A.; Westerink, B.H.C.; Verpoorte, E. Microdialysis-Coupled Enzymatic
Microreactor for in vivo Glucose Monitoring in Rats. Anal. Chem. 2013, 85, 10949–10955. [CrossRef] [PubMed]
23. Yu, B.; Long, N.; Moussy, Y.; Moussy, F. A long-term flexible minimally-invasive implantable glucose
biosensor based on an epoxy-enhanced polyurethane membrane. Biosens. Bioelectron. 2006, 21, 2275–2282.
[CrossRef] [PubMed]
24. Zhu, Z.G.; Song, W.H.; Burugapalli, K.; Moussy, F. Nano-yarn carbon nanotube fiber based enzymatic
glucose biosensor. Nanotechnology 2010, 21, 165501–165510. [CrossRef] [PubMed]
25. Zhu, Z.G.; Garcia-Gancedo, L.; Flewitt, A.J.; Milne, W.I.; Moussy, F. Design of carbon nanotube fiber
microelectrode for glucose biosensing. J. Chem. Technol. Biotechnol. 2012, 87, 256–262. [CrossRef]
26. Al-Sagura, H.; Komathia, S.; Khanb, M.A.; Gurekc, A.G.; Hassana, A. A novel glucose sensor using lutetium
phthalocyanine as redox mediator in reduced graphene oxide conducting polymer multifunctional hydrogel.
Biosens. Bioelectron. 2006. [CrossRef] [PubMed]
27. Zhu, Z.G.; Chen, C.; Zhu, X.R.; Xie, R.S.; Flewitt, A.J.; Milne, W.I. Effects of Ni Deposition on the Electrochemical
Properties of CNT/Ni Electrode and Its Application for Glucose Sensing. J. Nanosci. Nanotechnol. 2015, 15,
3196–3199. [CrossRef] [PubMed]
28. El-Ads, E.H.; Galala, A.; Atta, N.F. The effect of A-site doping in a strontium palladium perovskite and its
applications for non-enzymatic glucose sensing. RSC Adv. 2016, 6, 16183–16196. [CrossRef]
29. Kataoka, K.; Miyazaki, H.; Bunya, M.; Okano, T.; Sakurai, Y. Totally synthetic polymer gels responding to
external glucose concentration: Their preparation and application to on–off regulation of insulin release.
J. Am. Chem. Soc. 1998, 120, 12694–12695. [CrossRef]
30. Horkay, F.; Cho, S.H.; Tathireddy, P.; Rieth, L.; Solzbacher, F.; Magda, J. Thermodynamic analysis of the
selectivity enhancement obtained by using smart hydrogels that are zwitterionic when detecting glucose
with boronic acid moieties. Sens. Actuators B Chem. 2011, 160, 1363–1371. [CrossRef] [PubMed]
31. Gamsey, S.; Suri, J.T.; Wessling, R.A.; Singaram, B. Continuous glucose detection using boronic acid-substituted
viologens in fluorescent hydrogels: Linker effects and extension to fiber optics. Langmuir 2006, 22, 9067–9074.
[CrossRef] [PubMed]
32. Lee, M.C.; Kabilan, S.; Hussain, A.; Yang, X.; Blyth, J.; Lowe, C.R. Glucose-sensitive holographic sensors for
monitoring bacterial growth. Anal. Chem. 2004, 76, 5748–5755. [CrossRef] [PubMed]
33. Yang, X.; Pan, X.; Blyth, J.; Lowe, C.R. Towards the real-time monitoring of glucose in tear fluid: Holographic
glucose sensors with reduced interference from lactate and pH. Biosens. Bioelectron. 2008, 23, 899–905.
[CrossRef] [PubMed]
34. Liu, Y.; Zhang, Y.; Guan, Y. New polymerized crystalline colloidal array for glucose sensing. Chem. Commun.
2009, 1, 1867–1869. [CrossRef] [PubMed]
35. Li, Y.Y.; Zhou, S.Q. A simple method to fabricate fluorescent glucose sensor based on dye-complexed
microgels. Sens. Actuators B Chem. 2013, 177, 1363–1371. [CrossRef]
36. Zhang, X.J.; Gao, C.M.; Lu, S.Y.; Duan, H.G.; Jing, N.N.; Dong, D.; Shi, C.F.; Liu, M.Z. Anti-photobleaching
flower-like microgels as optical nanobiosensors with high selectivity at physiological conditions for continuous
glucose monitoring. J. Mater. Chem. B 2014, 2, 5452–5460. [CrossRef]
37. Rosenthal, S.J.; Chang, J.C.; Kovtun, O.; McBride, J.R.; Tomlinson, I.D. Biocompatible quantum dots for
biological applications. Chem. Biol. 2011, 18, 10–24. [CrossRef] [PubMed]
38. Schubert, K.; Khalid, W.; Yue, Z.; Parak, W.J.; Lisdat, F. Quantum-dot-modified electrode in combination with
NADH-dependent dehydrogenase reactions for substrate analysis. Langmuir 2010, 26, 1395–1400. [CrossRef]
[PubMed]
39. Tang, L.; Zhu, Y.; Yang, X.; Sun, J.; Li, C. Self-assembled CNTs/CdS/dehydrogenase hybrid-based
amperometric biosensor triggered by photovoltaic effect. Biosens. Bioelectron. 2008, 24, 319–323. [CrossRef]
[PubMed]
40. Tanne, J.; Schafer, D.; Khalid, W.; Parak, W.J.; Lisdat, F. Light-controlled bioelectrochemical sensor based on
CdSe/ZnS quantum dots. Anal. Chem. 2011, 83, 7778–7785. [CrossRef] [PubMed]
41. Zheng, M.; Cui, Y.; Li, X.; Liu, S.; Tang, Z.J. Photoelectrochemical sensing of glucose based on quantum dot
and enzyme nanocomposites. Electroanal. Chem. 2011, 656, 167–173. [CrossRef]
Sensors 2017, 17, 182 16 of 19

42. Alrouq, F.A.; Al-Masri, A.A.; AL-Dokhi, L.M.; Alregaiey, K.A.; Bayoumy, N.M. Study of the association of
adrenomedullin and basic-fibroblast growth factors with the peripheral arterial blood flow and endothelial
dysfunction biomarkers in type 2 diabetic patients with peripheral vascular insufficiency. J. Biomed. Sci. 2014,
21, 94–102. [CrossRef] [PubMed]
43. Rosenbloom, A.J.; Gandhi, H.R.; Subrebost, G.L. Microdialysis coupled with an embedded systems controller
and CMOS image sensor. In Proceedings of the 2009 Annual International Conference of the IEEE Engineering
in Medicine and Biology Society, Minneapolis, MN, USA, 3–6 September 2009; pp. 1230–1233.
44. Devadhasan, J.P.; Kim, S. Toward CMOS image sensor based glucose monitoring. Analyst 2012, 137,
3917–3920. [CrossRef] [PubMed]
45. Devadhasan, J.P.; Kim, S.; Choi, C.S. CMOS image sensors as an efficient platform for glucose monitoring.
Analyst 2013, 138, 5679–5684. [CrossRef] [PubMed]
46. Wang, P.Y.; Lu, M.S.C. CMOS thermal sensor arrays for enzymatic glucose detection. IEEE Sens. J. 2011, 11,
3469–3475. [CrossRef]
47. Wickramasinghe, Y.; Yang, Y.; Spencer, S.A. Current Problems and Potential Techniques in In Vivo Glucose
Monitoring. J. Fluoresc. 2004, 14, 513–520. [CrossRef] [PubMed]
48. Chou, Y.N.; Chang, Y.; Wen, T.C. Applying Thermosettable Zwitterionic Copolymers as General Fouling-
Resistant and Thermal-Tolerant Biomaterial Interfaces. ACS Appl. Mater. Interf. 2015, 7, 10096–10107.
[CrossRef] [PubMed]
49. Mohsen, D.S.; Pazuki, G.; Vossoughi, M. PEGylated silica-enzyme nanoconjugates: A new frontier in large
scale separation of α-amylase. Sci. Rep. 2015, 5, 18221. [CrossRef] [PubMed]
50. Gerritsen, M.; Kros, A.; Sprakel, V.; Lutterman, J.A.; Nolte, R.J.M.; Jansen, J.A. Biocompatibility evaluation of
sol-gel coatings for subcutaneously implantable glucose sensors. Biomaterials 2000, 21, 71–78. [CrossRef]
51. Kros, A.; Gerritsen, M.; Sprakel, V.; Lutterman, J.A.; Sommerdijk, N.; Jansen, J.A.; Nolte, R.J.M. Silica-based
hybrid materials as biocompatible coatings for glucose sensors. Sens. Actuators B Chem. 2001, 81, 68–75.
[CrossRef]
52. Jayakumar, R.; Prabaharan, M.; Nair, S.V.; Tamura, H. Novel chitin and chitosan nanofibers in biomedical
applications. Biotechnol. Adv. 2010, 28, 142–150. [CrossRef] [PubMed]
53. Jayakumar, R.; Prabaharan, M.; Kumar, R.T.S.; Nair, S.V.; Tamura, H. Biomaterials based on chitin and
chitosan in wound dressing applications. Biotechnol. Adv. 2011, 29, 322–337. [CrossRef] [PubMed]
54. Morais, J.M.; Papadimitrakopoulos, F.; Burgess, D.J. Biomaterials/Tissue Interactions: Possible Solutions to
Overcome Foreign Body Response. AAPS J. 2010, 12, 188–196. [CrossRef] [PubMed]
55. Quinn, C.A.P.; Conner, R.E.; Heller, A. Biocompatible, glucose-perme able hydrogel for in situ coating of
implantable biosensors. Biomaterials 1997, 18, 1665–1670. [CrossRef]
56. Chen, C.; Zhao, X.L.; Bao, H.; Liang, X.W.; Zhu, Z.G.; Zhu, Y.H. Polymerized Crystalline Colloidal Array
Photonic Crystal with Enhanced Mechanical Property. Chem. Lett. 2015, 44, 1566–1568. [CrossRef]
57. Heo, Y.J.; Takeucho, S. Towards Smart Tattoos: Implantable Biosensors for Continuous Glucose Monitoring.
Adv. Healthc. Mater. 2013, 2, 43–56. [CrossRef] [PubMed]
58. Yeh, C.C.; Venault, A.; Chang, Y. Structural effect of poly(ethylene glycol) segmental length on biofouling
and hemocompatibility. Polym. J. 2016, 48, 1–8. [CrossRef]
59. Wang, J. In vivo glucose monitoring: Towards ‘Sense and Act’ feedback-loop individualized medical systems.
Talanta 2008, 75, 636–641. [CrossRef] [PubMed]
60. Kotanen, C.N.; Moussy, F.J.; Carrara, S.; Guiseppi-Elie, A. Implantable enzyme amperometric biosensors.
Biosens. Bioelectron. 2012, 35, 14–26. [CrossRef] [PubMed]
61. Hickey, T.; Kreutzer, D.; Burgess, D.J.; Moussy, F. Dexamethasone/PLGA microspheres for continuous
delivery of an anti-inflammatory drug for implantable medical devices. Biomaterials 2002, 23, 1649–1656.
[CrossRef]
62. Heo, D.N.; Song, S.J.; Kim, H.J.; Lee, Y.J.; Ko, W.K.; Lee, S.J.; Lee, D.H.; Park, S.J.; Zhang, L.J.; Kang, J.Y.; et al.
Multifunctional hydrogel coatings on the surface of neural cuff electrode for improving electrode-nerve
tissue interfaces. Acta Biomater. 2016, 39, 25–33. [CrossRef] [PubMed]
63. Avula, M.; Jones, D.; Rao, A.N.; McClain, D.; McGill, L.D.; Grainger, D.W.; Solzbacher, F. Local release of
masitinib alters in vivo implantable continuous glucose sensor performance. Biosens. Bioelectron. 2016, 77,
149–156. [CrossRef] [PubMed]
Sensors 2017, 17, 182 17 of 19

64. Yu, J.Y.; Zhu, Z.G.; Chen, C.; Li, Z.H.; Chen, Y.X. The implantable glucose biosensor based on the spiral-type
Pt-Ir electrode. Chin. J. Sens. Actuators 2016, 29, 9–14.
65. Yu, D.H.; Sun, C.L.; Zheng, Z.Z.; Wang, X.L.; Chen, D.Y.; Wu, H.; Wang, X.Q.; Shi, F.X. Inner ear delivery of
dexamethasone using injectable silk-polyethylene glycol (PEG) hydrogel. Int. J. Pharm. 2016, 503, 229–237.
[CrossRef] [PubMed]
66. O’Donnell, C.; Efron, N. Diabetes and contact lens wear. Clin. Exp. Optom. 2012, 95, 328–337. [CrossRef]
[PubMed]
67. Mann, A.; Tighe, B. Contact lens interactions with the tear film. Exp. Eye Res. 2013, 117, 88–98. [CrossRef]
[PubMed]
68. Yao, H.F.; Shum, A.J.; Cowan, M.; Lähdesmäki, I.; Parviz, B.A. A contact lens with embedded sensor for
monitoring tear glucose level. Biosens. Bioelectron. 2011, 26, 3290–3296. [CrossRef] [PubMed]
69. Chu, M.X.; Miyajima, K.; Takahashi, D.; Arakawa, T.; Sano, K.; Sawada, X.; Kudo, H.; Iwasaki, Y.; Akiyoshi, K.;
Mochizuki, M.; et al. Soft contact lens biosensor for in situ monitoring of tear glucose as non-invasive blood
sugar assessment. Talanta 2011, 83, 960–965. [CrossRef] [PubMed]
70. Alexeev, V.L.; Sharma, A.C.; Goponenko, A.V.; Das, S.; Lednev, I.K.; Wilcox, C.S.; Finegold, D.N.; Asher, S.A.
High Ionic Strength Glucose-Sensing Photonic Crystal. Anal. Chem. 2003, 75, 2316–2323. [CrossRef] [PubMed]
71. Ben-Moshe, M.; Alexeev, V.L.; Asher, S.A. Fast Responsive Crystalline Colloidal Array Photonic Crystal
Glucose Sensors. Anal. Chem. 2006, 78, 5149–5157. [CrossRef] [PubMed]
72. Zhang, C.J.; Cano, G.G.; Braun, P.V. Linear and Fast Hydrogel Glucose Sensor Materials Enabled by Volume
Resetting Agents. Adv. Mater. 2014, 26, 5678–5683. [CrossRef] [PubMed]
73. Hu, Y.M.; Jiang, X.M.; Zhang, L.Y.; Fan, J.; Wu, W.T. Construction of near-infrared photonic crystal
glucose-sensing materials for ratiometric sensing of glucose in tears. Biosens. Bioelectron. 2013, 48, 94–99.
[CrossRef] [PubMed]
74. Zhang, J.; Zhu, J.; Akhter, K.F.; Thomas, A.A. Encapsulation of BSA within Gelatin Nanoparticles-laden
Biopolymer Film. Mater. Res. Soc. Symp. Proc. 2010. [CrossRef]
75. Amer, S.; Yousuf, M.; Siddqiui, P.Q.; Alam, J. Salivary glucose concentrations in patients with diabetes
mellitus-a minimally invasive technique for monitoring blood glucose levels. Pak. J. Pharm. Sci. 2001, 14,
33–37. [PubMed]
76. Chavez, E.M.; Borrell, L.N.; Taylor, G.W.; Ship, J.A. A longitudinal analysis of salivary flow in control subjects
and older adults with type 2 diabetes. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 2001, 91, 166–173.
[CrossRef] [PubMed]
77. Lopez, M.E.; Colloca, M.E.; Paez, R.G.; Schallmach, J.N.; Koss, M.A.; Chervonagura, A. Salivary characteristics
of diabetic children. Braz. Dent. J. 2003, 14, 26–31. [CrossRef] [PubMed]
78. Aydin, S. A comparison of ghrelin, glucose, alpha-amylase and protein levels in saliva from diabetics.
J. Biochem. Mol. Biol. 2007, 40, 29–35. [CrossRef] [PubMed]
79. Kim, J.; Imani, S.; de Araujo, W.R.; Warchall, J.; Valdes-Ramirez, G.; Paixao, T.R.L.C.; Mercier, P.P.; Wang, J.
Wearable salivary uric acid mouthguard biosensor with integrated wireless electronics. Biosens. Bioelectron.
2015, 74, 1061–1068. [CrossRef] [PubMed]
80. Yamaguchi, M.; Mitsumori, M.; Kano, Y. Noninvasively measuring blood glucose using saliva. IEEE Eng.
Med. Biol. Mag. 1998, 17, 59–63. [CrossRef] [PubMed]
81. Abikshyeet, P.; Ramesh, V.; Oza, N. Glucose estimation in the salivary secretion of diabetes mellitus patients.
Targets Ther. 2012, 5, 149–154.
82. Liu, C.; Sheng, Y.; Sun, Y.; Feng, J.; Wang, S.; Zhang, J.; Xu, J.; Jiang, D. A glucose oxidase-coupled DNAzyme
sensor for glucose detection in tears and saliva. Biosens. Bioelectron. 2015, 70, 455–461. [CrossRef] [PubMed]
83. Soni, A.; Jha, S.K. A paper strip based non-invasive glucose biosensor for salivary analysis. Biosens. Bioelectron.
2015, 67, 763–768. [CrossRef] [PubMed]
84. Zhang, W.; Du, Y.; Wang, M.L. On-chip highly sensitive saliva glucose sensing using multilayer films
composed of single-walled carbon nanotubes, gold nanoparticles, and glucose oxidase. Sens. Bio-Sens. Res.
2015, 4, 96–102. [CrossRef]
85. Lambert, J.L.; Borchert, M.S. Assessijng Blood Brain Barrier Dynamics or Identifying or Measuring Selected
Substances or Toxins in a Subject by Analyzing Raman Spectrum Signals of Selected Regions in the Eye.
U.S. Patent 6,574,501, 3 June 2003.
Sensors 2017, 17, 182 18 of 19

86. Lipson, J.; Bernhardt, J.; Block, U.; Freeman, W.R.; Hofmeister, R.; Hristakeva, M.; Lenosky, T.; McNamara, R.;
Petrasek, D.; Veltkamp, D. Requirements for calibration in naninvaive glucose monitoring by Raman
spectroscopy. J. Diabetes Sci. Technol. 2009, 3, 233–241. [CrossRef] [PubMed]
87. Bonato, P. Wearable sensors and systems. IEEE Eng. Med. Biol. Mag. 2010, 293, 25–36. [CrossRef] [PubMed]
88. Buller, M.; Welles, A.; Jenkins, O.C.; Hoyt, R. Extreme health sensing: The challenges, technologies, and
strategies for active health sustainment of military personnel during training and combat missions. Proc. SPIE
2010. [CrossRef]
89. Jia, W.; Bandodkar, A.J.; Valdes-Ramirez, G.; Windmiller, J.R.; Yang, Z.; Ramirez, J.; Chan, G.; Wang, J.
Electrochemical Tattoo Biosensors for Real-Time Noninvasive Lactate Monitoring in Human Perspiration.
Anal. Chem. 2013, 85, 6553–6560. [CrossRef] [PubMed]
90. Thomas, N.; Lahdesmaki, I.; Parviz, B.A. A contact lens with an integrated lactate sensor. Sens. Actuators
B Chem. 2012, 162, 128–134. [CrossRef]
91. Schabmueller, C.G.J.; Loppow, D.; Piechotta, G.; Schutze, B.; Albers, J.; Hintsche, R. Micromachined sensor
for lactate monitoring in saliva. Biosens. Bioelectron. 2006, 21, 1770–1776. [CrossRef] [PubMed]
92. Claver, J.B.; Miron, M.C.V.; Capitan-Vallvey, L.F. Disposable electrochemiluminescent biosensor for lactate
determination in saliva. Analyst 2009, 134, 1423–1432. [CrossRef] [PubMed]
93. Kim, J.; Valdes-Ramirez, G.; Bandodkar, A.J.; Jia, W.Z.; Martinez, A.G.; Ramirez, J.; Mercier, P.; Wang, J.
Non-invasive mouthguard biosensor for continuous salivary monitoring of metabolites. Analyst 2014, 139,
1632–1636. [CrossRef] [PubMed]
94. Arakawa, T.; Kuroki, Y.; Nitta, H.; Chouhan, P.; Toma, K.; Sawada, S.; Takeuchi, S.; Sekita, T.; Akiyoshi, K.;
Minakuchi, S.; et al. Mouthguard biosensor with telemetry system for monitoring of saliva glucose: A novel
cavitas sensor. Biosens. Bioelectron. 2016, 84, 106–111. [CrossRef] [PubMed]
95. Luchansky, M.S.; Bailey, R.C. High-Q Optical Sensors for Chemical and Biological Analysis. Anal. Chem.
2012, 84, 793–821. [CrossRef] [PubMed]
96. Paek, S.H.; Cho, I.H.; Kim, D.H.; Jeon, J.W.; Lim, G.S.; Paek, S.H. Label-free, needle-type biosensor for
continuous glucose monitoring based on competitive binding. Biosens. Bioelectron. 2013, 40, 38–44. [CrossRef]
[PubMed]
97. Cummins, B.; Garza, J.; Cote, G. Optimization of a Concanavalin A-Based Glucose Sensor Using Fluorescence
Anisotropy. Anal. Chem. 2013, 85, 5397–5404. [CrossRef] [PubMed]
98. Shen, P.F.; Xia, Y.S. Synthesis-Modification Integration: One-Step Fabrication of Boronic Acid Functionalized
Carbon Dots for Fluorescent Blood Sugar Sensing. Anal. Chem. 2014, 86, 5323–5329. [CrossRef] [PubMed]
99. Srinivasan, G.; Chen, J.; Parisi, J.; Bruckner, C.; Yao, X.D.; Lei, Y. An Injectable PEG-BSA-Coumarin-GOx
Hydrogel for Fluorescence Turn-on Glucose Detection. Appl. Biochem. Biotechnol. 2015, 5, 1115–1126. [CrossRef]
[PubMed]
100. Srivastava, R.; Jayant, R.D.; Chaudhary, A.; Mcshane, M.J. “Smart Tattoo” Glucose Biosensors and Effect of
Coencapsulated Anti-inflammatory Agents. J. Diabetes Sci. Technol. 2011, 1, 76–85. [CrossRef]
101. Wang, H.C.; Lee, A.R. Recent developments in blood glucose sensors. J. Food. Drug. Anal. 2015, 2, 191–200.
[CrossRef]
102. Ferrari, M.; Quaresima, V. A brief review on the history of human functional near-infrared spectroscopy
(fNIRS) development and fields of application. Neurolmage 2012, 63, 921–935. [CrossRef] [PubMed]
103. So, C.F.; Choi, K.S.; Wong, T.K.S.; Chung, J.W.Y. Recent advances in noninvasive glucose monitoring.
Med. Devices 2012, 5, 45–52.
104. Vashist, S.K. Non-invasive glucose monitoring technology in diabetes management: A review. Anal. Chim. Acta
2012, 750, 16–27. [CrossRef] [PubMed]
105. Hazen, K.H.; Arnold, M.A.; Small, G.W. Measurement of glucose in water with first-overtone near-infrared
spectra. Appl. Spectrosc. 1998, 52, 1597–1605. [CrossRef]
106. Genina, E.A.; Bashkatov, A.N.; Sinichkin, Y.P.; Yanina, I.Y.; Tuchin, V.V. Optical clearing of biological tissues:
Prospects of application in medical diagnostics and phototherapy. J. Biomed. Photonics Eng. 2015, 1, 24–58.
[CrossRef]
107. Stiles, P.L.; Dieringer, J.A.; Shah, N.C.; Van Duyne, R.P. Surface-enhanced Raman spectroscopy. Anal. Chem.
2008, 1, 601–626. [CrossRef] [PubMed]
Sensors 2017, 17, 182 19 of 19

108. Torul, H.; Ciftci, H.; Dudak, F.C.; Adiguzel, Y.; Kulah, H.; Boyaci, I.H.; Tamer, U. Glucose determination
based on a two component self-assembled monolayer functionalized surfaceenhanced Raman spectroscopy
(SERS) probe. Anal. Methods 2014, 6, 5097–5104. [CrossRef]
109. Kong, K.V.; Ho, C.J.H.; Gong, F.X.; Lau, W.K.O. Sensitive SERS glucose sensing in biological media using
alkyne functionalized boronic acid on planar substrates. Biosens. Bioelectron. 2014, 56, 186–191. [CrossRef]
[PubMed]
110. Harris, J.M.; Reyes, C.; Lopez, G.P. Common causes of glucose oxidase instability in in vivo biosensing:
A brief review. J. Diabetes Sci. Technol. 2013, 7, 1030–1038. [CrossRef] [PubMed]

© 2017 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).