R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q.

, 29 (4) 511–518 (2015) 511

Rheological Studies Dedicated to the Development

of a Novel Injectable Polymeric Blend for Viscosupplementation Treatment
R. Lapasin,a F. Segatti,b D. Mercuri,b G. De Conti,b C. Spagnul,b and S. Fusib
a
Department of Engineering and Architecture, University of Trieste, doi: 10.15255/CABEQ.2014.2148
via A. Valerio 6/A, I-34127 Trieste, Italy
b
R&D Department, Limacorporate S.p.a., via Nazionale 52, Original scientific paper
I-33038 Villanova di San Daniele del Friuli, Italy Received: November 15, 2014
Accepted: November 30, 2015

Viscosupplementation is an intra-articular symptomatic treatment of mild osteoar-

thritis. The treatment involves the injection of high-molecular-weight hyaluronan (HA),
and especially of cross-linked HA to restore the lubricating and cushioning properties of
the synovial fluid.
This work involves the development of a novel viscosupplementation fluid based on
amidated carboxymethylcellulose and obtained by blending the soluble polymer with its
crosslinked derivative. Rheological analyses carried out under both oscillatory and con-
tinuous shear provided a rationale to assess the viscosupplement formulation and the
production process. The hydrogel fraction content and the total polymer concentration
can be properly selected in order to ensure an optimal combination of flowability and
viscoelastic properties.
Key words:
amidated carboxymethylcellulose, viscosupplementation, polymeric blend, hydrogel,
rheology

Introduction OARSI treatment guidelines.22 Viscosupplementa-

tion consists in intra-articular, single or repeated,
Osteoarthritis (OA) is a chronic, progressive injections of HA or HA-based derivatives (HAs).
musculoskeletal disorder1–3 that mainly affects the Adequate viscoelastic properties are also important
weight-bearing chondro-synovial joints such as to restore the articular homeostasis.23 HA or HAs
knee, hip4 and the upper extremities.5,6 The disease increase the elastoviscosity of the intercellular ma-
affects the ageing population worldwide7,8 leading trix creating a mechanical protective socket that
to a high socio-economic burden9 with substantial surrounds nociceptive nerve endings. This results in
financial consequences.10,11 a decrease in pain and an increase in joint mobility24
In a healthy condition, the lubricants present in as it has already been demonstrated on animals
the synovial fluid, such as Hyaluronic acid (HA), models.25,26
lubricin12 and the surface-active phospholipids One of the most important drawbacks of HA
(SAPL)13,14 act as shock absorbers protecting the and HAs is the multiple injection therapy due to the
joint against compressive damage.15–17 Hyaluronic enzymatic degradation by hyaluronidases in the
acid (HA) is responsible for the viscoelastic proper- body. High molecular weight products and cross-
ties.18 linked hydrogels have been developed, but they are
In OA joint diseases, the concentration and mo- able to retard the hyaluronidases attack without pre-
lecular weight of HA decrease16,19 and the synovial venting it.27 Furthermore, inflammatory reactions
fluid becomes more abundant, less viscous and re- sometimes accompanied by swelling and erythema
duced in elastoviscous properties that may culmi- have also been variably reported.28,29
nate in a painful condition for the patient. A new derivative similar to HA, but with a bet-
Viscosupplementation20,21 was identified as an ter behaviour in terms of hardness and stability to-
effective treatment of OA with a small number of wards enzymatic degradation30 has been developed
adverse effects. Nowadays, it is approved in Eu- converting 50 % of the carboxylic groups of car-
rope/USA for the treatment of pain associated with boxymethylcellulose (CMC) into amidic groups to
OA of the knee, but considerable controversy about obtain the amidated carboxymethylcellulose
its efficacy is still ongoing as evidenced in the (CMCA).31 In vitro results showed that the amida-
tion significantly ameliorated the biocompatibility
Corresponding author: [email protected]
*
of the material and diminished the cell expression
512 R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q., 29 (4) 511–518 (2015)

of inflammatory cytokines. CMCA also positively molar ratio with the polymer was 0.5:1 to obtain a
influenced osteoblast activity.30 The vegetal origin crosslinking degree of about 50 %. The obtained
of CMCA potentially avoids the risk of allergic re- hydrogel was purified by repeated rinsing in puri-
actions due to certain protein components present in fied water and freeze dried. The two freeze-dried
the HA and Ha-based products injected. components, polymer and hydrogel, were milled in
This work involves the development of a novel order to have a controlled cut off, up to 2 mm, that
viscosupplementation fluid obtained by blending were subsequently blended and hydrated in saline
the soluble CMCA polymer (hereinafter indicated solution, and eventually filled in syringe and steril-
as CMCAp) with its crosslinked hydrogel deriva- ized.
tive (CMCAg) by crosslinking the residual 50 %
fraction of CMCAp carboxylic groups. Rheological experiments
CMCA-based fluids were investigated to assess Rheological characterization was carried out
the rheological properties of the viscosupplement with a rotational controlled stress rheometer
fluid during the different processing stages. Both (Rheostress RS150, Haake GmbH, Germany),
continuous and oscillatory shear tests showed that, equipped with a cone/plate device (d = 60 mm, an-
at equal polymer concentration, aqueous CMCAp gle = 1°) or a parallel plate geometry (d = 35 mm,
and CMCAg systems exhibit quite different rheo- gap = 1 mm) with serrated surfaces in order to avoid
logical behaviours, typical of polymer solutions and wall slippage effects. The apparatus was comput-
gels, respectively, and hence a wide arc of viscous er-controlled (RheoWin software program) and the
and viscoelastic responses can be obtained by temperature was kept at 25 ± 0.1 °C (Thermostat
blending the two polymers in different proportions. TC500, Haake GmbH, Germany). The flow proper-
The gel fraction (CMCAg/CMCAp ratio) and the ties were determined applying a logarithmic se-
total polymer concentration of the blend can be ap- quence of increasing constant stresses in the range
propriately selected in order to ensure an optimal 1-1000 Pa and measuring the corresponding shear
combination of viscoelastic properties and flow be- rate. Each stress value was kept constant until the
haviour (injectability) together with an acceptable corresponding time variation of the shear rate satis-
structural recovery after shear. fied the pre-set constraint for steady state attain-
ment, or at most for 90 s.
Materials and methods Oscillatory stress sweep tests were performed
at 1 Hz in order to individuate the extension of the
Materials linear viscoelasticity regime. All the frequency
sweep measurements were performed from 100 to
The raw material for CMCA preparation is so- 0.01 Hz within the linear range.
dium carboxymethylcellulose (average molecular
Recovery tests were performed in order to
weight: 750 kDa, substitution degree 0.8) (CP
evaluate the structural recovery after the application
Kelko). It can be functionalized thanks to the pres-
of continuous shear conditions. They were com-
ence of two carboxyl groups in the polymeric unit
posed of three consecutive segments, alternatively
through a water-based synthetic pathway. The car-
under small amplitude oscillatory conditions (at 1
boxylic groups are first activated with NHS/EDC
Hz) and in continuous shear at constant shear rate
(N-hydroxysuccinimide purity >97 % (Sigma-Al-
(5 s–1, 180 s). The steady values of G’ and G” at-
drich); N-(3-dimethylaminopropyl)-N’-ethylcarbo-
tained in the oscillatory segments before and after
diimide hydrochloride purity >99 % (Actim)). Once
shear flow were compared.
activated, they quickly react with methylamine
(40 % water solution, Sigma-Aldrich), molar ratio All the experimental tests were done in dupli-
of 1:1. The pH of the solution is always kept at cate or triplicate.
­4.5–5 adjusting hydrochloric acid. The amide-CMCA
was then purified in a buffered solution (about pH Data treatment and modeling
8–8.5 in order to increase the solubility of the poly-
The experimental viscosity data can be cor-
mer) through a Tangential Flow Filtration instru-
related quite satisfactorily with the Carreau-Yasuda
ment using a membrane with 100 kDa cut off. The
model:32,33
obtained solution was separated into two aliquots:
the former was freeze-dried, the latter was used for η0 − η∞
the crosslinking process. η = η∞ +
[1+ ( a γ )b ] p
The amide-CMC polymer is crosslinked by a
solution of NHS/EDC and a bi-functional group. In where h0 and h∞ represent the zero-shear rate vis-
particular, 1,3-diaminopropane (purity 99.0 %, Sig- cosity and the infinite-shear rate viscosity, respec-
ma-Aldrich) was used as crosslinking agent. The tively. In this study, the high shear Newtonian pla-
R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q., 29 (4) 511–518 (2015) 513

teau is beyond the experimental window and, hence,

h∞ was arbitrarily set equal to 0.89 mPa s, i.e. to the
water viscosity at 25 °C. The product bp rules the
slope of the flow curve in the power law region,
while the critical shear rate gc marking the onset of
shear thinning is strictly related to the characteristic
time a.
The mechanical spectra can be described with
the generalized Maxwell model through the follow-
ing expressions:
N N
G ω2 λ 2 G ωλ
G′= ∑ i 2 i 2 G′′= ∑ i 2 i 2
i=1
1+ ω λi i=1
1+ ω λi

where Gi and li are the modulus and the relaxation

time of the ith Maxwell element, respectively. In or-
der to reduce the problem related to parameter cor-
relation, a scaling criterion was adopted for the re- F i g . 2 – Mechanical spectra of aqueous CMC and CMCAp
laxation time sequence (li+1 = 10 li) and not more 1 % w/w solutions
than 5 Maxwell elements were taken into consider-
ation for data fitting. through the variations of two model parameters: the
zero shear rate viscosity h0 and the critical shear
rate gc.
Results and discussion
The changes induced by amidation on the lin-
The effects produced by amidation on the rheo- ear viscoelastic behaviour of solutions were coher-
logical behaviour of CMC solutions appeared to be ent with those observed for their flow properties. At
more evident at low shear conditions and higher equal polymer concentration, the mechanical spec-
polymer concentrations. Comparing the flow curves trum moved towards lower values of both moduli
of CMC and CMCAp solutions at equal concentra- and the crossover point shifted to higher frequen-
tion, the zero-shear-rate viscosity diminished after cies because of the faster relaxation modes. The ex-
amidation and the Newtonian plateau became more ample reported in Fig. 2 regards the 1 % solutions.
extended towards higher shear conditions, as it can These effects can be evaluated more precisely by
be observed in Fig. 1. Similar rheological effects of comparing the relaxation time spectra obtained
amidic moieties on carboxymethylcellulose have from data correlation with the Maxwell generalized
been observed by Barbucci et al.30 model, as illustrated in Fig. 3.
A satisfactory data correlation was provided by More drastic changes were produced by poly-
the Carreau-Yasuda equation. Accordingly, the mer crosslinking, particularly in linear viscoelastic
above underlined effects can be concisely described conditions; this can be seen by comparing the re-

F i g . 1 – Steady viscosity curves of aqueous CMC and CMCAp F i g . 3 – Relaxation time spectra of aqueous CMC and CMCAp
solutions at different concentrations (expressed in 1 % w/w solutions derived from data correlation
% w/w) with the Maxwell generalized model
514 R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q., 29 (4) 511–518 (2015)

sions). For CMCAg systems, the critical strain

marking the borderline of the linear viscoelastic re-
gime became sensibly lower (two orders of magni-
tude or more) than for CMCAp solutions.
The quite different profiles of the mechanical
spectra of CMCAp and CMCAg systems reflect the
neat changes produced by crosslinking in linear vis-
coelastic properties and relaxation modes, thus evi-
dencing the structural transition from solution to
physical gel (Fig. 4b). In the experimental window
explored, the behaviour of the CMCAp solution
was characterized by a prevailing viscous compo-
nent (G”>G’) and short relaxation times, as it can
be deduced by the crossover frequency. In the case
of the crosslinked polymer, the storage modulus
curve is placed clearly above G” values, and reach-
es a plateau condition at low frequencies, where the
divergence from the linear viscoelastic responses of
CMCAp is quite evident.
Studying the steady flow properties, we found
that CMCAg systems exhibited much more pro-
nounced shear thinning than CMCAp solutions, ow-
ing to their different structural conditions and
shear-induced mechanisms. CMCAg systems be-
have as dispersions of gel microdomains, which
may undergo significant deformations and spatial
rearrangements with increasing shear rate.
Looking at the problems connected to the de-
velopment of a novel polymeric fluid for visco­
supplementation treatment, the first decisive step
concerns its formulation in relation to the structural
and rheological properties that the fluid must pos-
sess and exhibit. In particular, it should be easily
F i g . 4 – Viscoelastic responses of 2.5 % CMCAp and 2.5 % injectable and display appropriate viscoelastic re-
CMCAg systems; a) storage and loss moduli vs strain (from sponses ‘in situ’ in the various kinematic condi-
stress sweep test); b) mechanical spectra (from frequency tions experienced by articular joints. Furthermore,
sweep test) an adequate resistance to enzymatic degradation
and mechanical solicitations for prolonged times
sponses of both of CMCAp and CMCAg systems should be ensured. Blending CMCAp solutions
with the same concentration of soluble and cross- and CMCAg microgel dispersions it is possible to
linked polymer (2.5 % for systems represented in finely adjust the composition ratio and the total
Figs. 4a and 4b, respectively). polymer concentration in order to modulate the
Stress sweep tests showed that both viscoelas- structural features and rheological properties of
tic moduli of the CMCAg system were markedly both systems.
higher, and in the nonlinear region, their profiles The following figures illustrate how the rheo-
were different from those of the corresponding CM- logical properties of CMCAp-CMCAg blends
CAp solutions (Fig. 4a). For the latter systems, the changed with increasing gel fraction at equal total
transition from linear to nonlinear regime is de- polymer concentration. The steady viscosity strong-
scribed by monotonic decrease of both moduli. This ly increased at low shear rates, while its change was
behaviour is usually observed in ordinary polymeric almost negligible at high shear rates. As the gel
solutions and labelled as type I according to the fraction increased, the shear thinning character of
classification of LAOS responses.34 Conversely, in the flow behaviour became more and more marked,
the case of CMCAg systems, the storage modulus being ruled by the level and extension of the New-
decreased monotonically with increasing strain tonian plateau (Fig. 5).
whereas a weak overshoot appeared in the G’’ pro- The experimental data can be correlated quite
file. Such a pattern (Type III) is typical of weakly satisfactorily with the Carreau-Yasuda equation.
structured fluids (physical gels or colloidal disper- The effects of increasing gel fraction fg on viscous
R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q., 29 (4) 511–518 (2015) 515

F i g . 5 – Steady viscosity curves of 2.5 % CMCAp-CMCAg

blends at different gel fractions (1 represents the polymer/gel
ratio of 0/10; 0 represents a polymer/gel ratio of 10/0)

properties of CMCAp-CMCAg blends can be rea-

sonably described through the only variations of the
zero-shear rate viscosity h0 and the critical shear F i g . 7 – Mechanical spectra of 2.5 % CMCAp-CMCAg
rate gc, marking the border of the Newtonian plateau blends at different gel fractions (0 represents the polymer/gel
(Fig. 6). Thus, it can be noted that the most signifi- ratio of 10/0; 1 represents a polymer/gel ratio of 0/10)
cant changes occurred in the fg interval between 0.2
and 0.3, where gc drops significantly, and the rate of served in the shape of the mechanical spectrum,
h0 increase became slower. At higher fg values, the passing from CMCAp solution to CMCAg system.
increasing concentration of gel microdomains in the According to the Winter-Chambon criterion35,36, the
whole polymeric phase resulted only in an exponen- sol-gel transition could be located near fg = 0.3,
tial h0 increase with no appreciable effect on the since the traces of G’ and G’’ of system 7 were
critical shear rate. ­almost coincident and obey the same scaling law
Similarly, the linear viscoelastic behaviour of (G’    G’’    wn, n    0.45) in the experimentally ex-
the blend significantly changed with increasing gel plored frequency window.
fraction. To clarify, the graphical representation re- The generalized Maxwell model provides a sat-
ported in Fig. 7 involves only a partial subset of the isfactory data correlation and, consequently, the re-
mechanical spectra experimentally determined. As sulting relaxation time spectra can be profitably
may be observed, the increasing gel fraction fg led used for analysing the blending effects on the linear
to an increase in the elastic character of the visco- viscoelastic responses in the place of the mechani-
elastic response, which was more pronounced at cal spectra (Fig. 8). As the CMCAg content of the
lower frequencies where the storage modulus in- polymeric phase increased, the relative weight of
creased more markedly than the viscous compo- the slowest relaxation modes became more import-
nent. Consequently, a progressive change was ob-

F i g . 8 – Relaxation time spectra derived from data correlation

with the generalized Maxwell model for 2.5 % CMCAp-CMCAg
F i g . 6 – Zero-shear rate viscosity and critical shear rate vs blends (1 represents the polymer/gel ratio of 0/10, 0 represents
gel fraction in 2.5 % CMCAp-CMCAg blends a polymer/gel ratio of 10/0)
516 R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q., 29 (4) 511–518 (2015)

ant owing to the increasing contribution of gel mi-

crodomains towards a structural state close to jam-
ming.
The analysis of the reversibility of linear visco-
elastic responses (G’, G’’) can be particularly useful
in view of the development of a novel viscosupple-
ment, which must be injected, and hence undergoes
short but intense shear conditions in the application
phase. Recovery tests performed on CMCAp-CMCAg
blends showed that the viscoelastic behaviour was
completely reversible for CMCAp solution, since
the values of G’ and G” before and after shear flow
were shown to be equal. On the other hand, only a
partial recovery was attained for CMCAg system F i g . 9 – Reduction ratio of viscoelastic moduli between unster-
along the time segment following the application of ile and sterile 2.5 % CMCAp-CMCAg blends vs gel
a constant shear rate. The viscoelastic recovery de- fraction at two reference frequencies (0.5 and 2.5 Hz)
creased with increasing gel fraction.
Investigations were also carried out on the de- manufacturing process, in particular that caused by
velopment of a pilot production process and evalu- steam sterilization.
ations were made on the potential effect of each Typical moist-heat sterilization cycles, based
processing step on the blend’s final properties. on exposition to temperature of 121 °C for 15’ have
Standard production implies a set of consecutive been applied, and the mechanical spectra obtained
steps to obtain both components of the polymeric for unsterile and sterile blends of equal total poly-
blend: CMC solubilization, CMC functionalization mer concentration and gel fraction have been com-
in order to obtain CMCA, purification of CMCA, pared. As highlighted in Fig. 9, it can be seen that
concentration and freeze-drying of the polymer, steam sterilization induced a loss of both elastic and
crosslinking of the CMCA, purification of the ob- viscous moduli due to the polymer degradation, and
tained hydrogel, concentration and freeze-drying of hence a reduction in the main properties of the de-
the hydrogel. The final steps cover rehydration in vice.
saline solution, polymer and gel blending, filling in Consequently, the initial formulation must be
syringe and sterilization. readjusted in terms of total polymer concentration
Among the several steps of the manufactur- and gel fraction in order to compensate the negative
ing process, some of them can strongly affect the effects produced by the manufacturing process on
elastoviscosity of the polymer blend. It has been the rheological behaviour of the blend and to re-
observed that freeze drying, homogenization in
­ store a correct balance among viscoelasticity, stabil-
­saline solution and filling in syringes can produce ity and easy injectability. In fact rheological proper-
a drastic decrease in loss and storage moduli in ties of numerous viscosupplements available on the
both soluble and crosslinked CMCA. Moreover, market cover a wide spectrum of values and some-
because viscosupplements are classified as phar­
­ times they are quite different from those peculiar to
maceutical products or medical devices, an essen- healthy synovial fluids.20,38–40
tial requirement for CMCA blends is represent- This suggests that the main requirements for
ed by their capability to resist the sterilization such devices are represented by their viscoelastic
process. properties in order to exhibit a proper response to
Several techniques based on chemical agents the stress conditions within synovial joints, together
(EtO, plasma) or physical processes (heating, b or γ with their capability to be easily injected through an
radiation) are widely used for sterilization in the 18-20 G needle. It is also acknowledged that the
pharmaceutical and medical devices industries. values of the linear viscoelastic moduli measured at
Among them, the most suitable and promising 0.5 and 2.5 Hz can represent a suitable way to eval-
method in terms of efficacy and compatibility with uate how synovial fluid and viscosupplements react
solution or emulsions is represented by steam or to walking and running conditions, respectively.20,41
moist heat. Conversely, it is known that heat induc- Accordingly, they can serve to discriminate the be-
es degradation of CMC gels and polymer solutions37 haviours of synovial fluids in normal and patholog-
and at high temperatures CMC degrades and perma- ical conditions, as well as to assess the effects of
nently reduces in viscosity. Therefore, a rheological viscosupplementation therapies.39,40,42 The experi-
investigation has been dedicated to evaluate the po- mental values obtained for a series of commercial
tential degradation of CMCA blends during the viscosupplements are reported in Fig. 10a. The
R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q., 29 (4) 511–518 (2015) 517

quite large interval covered by both moduli at both

frequencies is strictly related to the different me-
chanical spectra displayed by the systems consid-
ered, spanning from those typical of ordinary poly-
meric solutions to classical gel behaviours. Similar
differences can also be found among novel systems
proposed as candidates for viscosupplementa-
tion.40,42–45
Tests performed on blends with different gel
fractions have shown that an acceptable structural
recovery after continuous shear application (more
than 80–90 %) is attained when the gel fraction is
lower than 50 %. Thus, the 6/4 CMCAp/CMCAg
blend has been provisionally selected for the final
investigation addressed to individuate the most
­suitable total polymer concentration, apt to compen-
sate the loss in mechanical properties occurring
during processing and sterilization. The viscoelastic
moduli of three 6/4 CMCAp/CMCAg blends with
different total polymer concentrations are compared
with those of two commercial products in Fig. 10b,
while Fig. 10c reports the viscosity values of the
same systems at three different shear rates. From
both figures, it can be inferred that the 3.61 w/w
concentration provides adequate viscoelastic prop-
erties, which are combined with a proper injecta­
bility.

Conclusions

Blends with tailored viscoelastic properties can

be obtained from CMCA in the form of polymer
solutions and crosslinked hydrogels.
More specifically, the entire range between
F i g . 1 0 – a: Viscoelastic moduli (at 0.5 and 2.5 Hz) of the 10/0 and 0/10 of polymer/hydrogel ratio has been
commercial viscosupplements listed below. b: Viscoelastic moduli investigated, and the 5/5-6/4-7/3 polymer/hydrogel
(at 0.5 and 2.5 Hz) of the 6/4 CMCAp/CMCAg blends at 2.5 %, compositions seem to create a behaviour very close
3 % and 3.61 % w/w concentrations compared with those of two to the main characteristics of an HA-based product
commercial products (Orthovisc and Monovisc). c: Viscosity
values (at 0.1, 10 and 1000 s–1) of the 6/4 CMCAp/CMCAg blends for viscosupplementation.
at 2.5 %, 3 % and 3.61 % w/w concentrations compared with This study has demonstrated the importance of
those of two commercial products (Orthovisc and Monovisc). both polymer and hydrogel form in blends, because,
Commercial viscosupplement
Suplasyn (Mylan N.V., UK), non-heat degraded highly purified sodium if singularly used, (10/0 and 0/10 of polymer/hydro-
hyaluronate (500 to 730 kDa), measured values, ref. 39.; Hyalubrix (Fidia gel samples), they are unable to preserve both stor-
farmaceutici S.p.A., Italy), sodium hyaluronate (1300 to 3600 kDa), from
bacteria fermentation, estimated values, ref. 40.; Orthovisc (Anika Ther-
age and loss moduli.
apeutics MA, USA), highly purified sodium hyaluronate in physiologic Properties, such as lubrication and shock ab-
saline, measured values, ref. 39.; Monovisc (Anika Therapeutics MA,
USA) highly purified, partially crosslinked sodium hyaluronate in a phos- sorbing, together with an adequate resistance to
phate buffered saline solution, estimated values, ref. 40.; Noltrex (JSC processing and steam sterilization, can be assured
“Research center “Bioform”, Russia), synthetic polyacrylamide hydrogel by using a concentration of 3.61 % w/w.
with silver ions, measured values in our laboratory; Hymovis (Fidia far-
maceutici S.p.A., Italy), partially hydrophobized derivative of hyaluronic The possibility to formulate CMCA in several
acid, estimated values, ref. 40.; Synvisc (Hylan G-F 20) (Genzyme Bio-
surgery, USA). blend 80/20 of Hylan A (high molecular weight hyaluron-
polymer/hydrogel blends and in different concentra-
ic acid) and Hylan B (crosslinked hyaluronic acid), measured values, ref. tions, together with its biocompatibility and its slow
39.: Durolane (Bioventus, USA), crosslinked hyaluronic acid, measured enzymatic degradation, should be considered as a
values in our laboratory.; Fermathron (Hyaltech Ltd. UK), sodium hyal-
uronate (1300 to 3600 kDa) in a phosphate buffered saline solution, from promising alternative to HA for the design of new
bacterial fermentation, measured values in our laboratory. products for viscosupplementation.
518 R. LAPASIN et al., Rheological Studies Dedicated to the Development of a Novel…, Chem. Biochem. Eng. Q., 29 (4) 511–518 (2015)

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