Anda di halaman 1dari 7

Materials Science and Engineering C 73 (2017) 333339

Contents lists available at ScienceDirect

Materials Science and Engineering C

journal homepage: www.elsevier.com/locate/msec

Development and characterization of tripolymeric and bipolymeric


composite lms using glyoxal as a potent crosslinker for
biomedical application
Ashwini Kumar, Awanish Kumar
Department of Biotechnology, National Institute of Technology, G. E. Road, Raipur 492010, Chhattisgarh, India

a r t i c l e i n f o a b s t r a c t

Article history: For various biomedical applications, biopolymeric lms are often crosslinked using chemical crosslinker such as
Received 26 August 2016 glutaraldehyde, which is considered as a toxic chemical. In this report, we have prepared and characterized
Received in revised form 7 December 2016 biopolymeric lms using different combinations of chitosan, CMC, alginate and PVA using glyoxal as the
Accepted 13 December 2016
crosslinker. The prepared lms were subjected to various physico-chemical and mechanical characterizations
Available online 19 December 2016
such as swelling index, surface pH, surface morphology analysis using SEM, interact ion study using XRD, exibil-
Keywords:
ity study using tensile testing and hardness testing. Glyoxal crosslinking resulted in variation of physico-chemical
Alginate and mechanical alteration of the chitosan-PVA lms while it had the negligible effect on the CAP lm. Further, the
Chitosan hardness of the lms demonstrated a decrease in value in the crosslinked lms as compared to non-crosslinked
CMC lms. We have interpreted that glyoxal is a potential crosslinker for chitosan-based composite polymers while in
Glyoxal this case, it did not show any signicant effect on CMC and alginate based composite structures. Therefore, using
PVA this type of lms would be the cheap, safe and new alternative in drug delivery and other biomedical
Biomedical applications applications.
2016 Elsevier B.V. All rights reserved.

1. Introduction natural bioadhesive too. Being a cationic polymer, it interacts with an-
ionic groups easily and forms a strong bond. A few examples of its
Biopolymers have been a boon for various biomedical applications bioadhesive application are its mucoadhesive drug delivery system
such as drug delivery, tissue adhesives, wound healing coverings and and bioadhesive surgical sutures [3]. Some other natural and semi-nat-
scaffolds for tissue engineering. They are being employed attributed to ural mucoadhesive polymers are alginate (ALG), carboxymethyl cellu-
their properties of biodegradability, biocompatibility, adhesiveness lose (CMC), Hydroxypropyl methyl cellulose (HPMC) while poly
and non-immunogenicity. Various natural, semi-natural and synthetic acrylic acid is the most utilized synthetic mucoadhesive polymer [4].
polymers such as chitosan, silk, alginate, cellulosic polymers, poly acryl- Mucoadhesive biopolymeric blended lms are being clinically and ex-
ic acid (PAA), poly-lactic acid (PLA), poly (lactic-co-glycolic acid) perimentally tested for transbuccal and transdermal drug delivery sys-
(PLGA), polycaprolactone (PCL) have been employed for various appli- tems. Transbuccal delivery of drugs primarily depends upon the
cations. Natural and semi-synthetic biopolymers have added advantage interaction of the polymer and the mucin protein of the mucous mem-
of being readily biodegradable and a few are bioadhesive too and there- brane, permeation enhancing effect of added permeation enhancer and
fore they are widely utilized for drug delivery applications [1,2]. the swelling behaviour of the formulated lms [5]. Though
Chitosan, the deacetylated version of the parent polymer chitin, is mucoadhesive polymers specically do not show adhesiveness over
one of the most exploited biopolymers. Apart from being biodegradable skin, the formulated patches often are backed by an adhesive layer
and biocompatible, it has a great anti-microbial activity. Chitosan is a which could hold back the patch over skin for transdermal application.
The polymeric lms are also incorporated with special permeation en-
hancers which could make it easier for the drug to permeate through
the highly keratinized epidermis [6]. Wound healing covering is another
Abbreviations: ASTM, American Standard Testing Materials; CMC, carboxy methyl wide area of application of polymers. Being a great anti-microbial com-
cellulose; HPMC, hydroxypropyl methyl cellulose; IS, Indian standard; NaOH, sodium pound, chitosan is being used alone or as the primary ingredient of var-
hydroxide; PVA, poly vinyl alcohol; PAA, poly acrylic acid; PLA, poly lactic acid; PLGA,
poly lactic-co-glycolic acid; SEM, scanning electron microscopy; XRD, X-ray diffraction.
ious patches used as wound covering [7]. A lot of in vitro, preclinical and
Corresponding author. clinical studies have been reviewed by Dai et al. regarding the antimi-
E-mail address: drawanishkr@gmail.com (A. Kumar). crobial and wound healing effect of chitosan based formulations [8].

http://dx.doi.org/10.1016/j.msec.2016.12.061
0928-4931/ 2016 Elsevier B.V. All rights reserved.
334 A. Kumar, A. Kumar / Materials Science and Engineering C 73 (2017) 333339

Fig. 1. A brief pictorial demonstration of solvent casting method.

Very often, blended polymeric lms are crosslinked during their for- used as the plasticizer for all the formulations. Glyoxal (5% v/v) was
mulation to impart them strength. The most common chemical used as the crosslinker for 1 formulation each of both sets. The individ-
crosslinker applied is glutaraldehyde. Glutaraldehyde, however, is con- ual ingredients were mixed thoroughly on a magnetic stirrer and were
sidered toxic for respiratory tract, eyes and skin and therefore alterna- further mixed under a homogenizer to get a clear uniform solution.
tive cross-linkers are being utilized for the polymeric lms [9]. A few The solution was further kept overnight to get degassed. The lms
works has reported that glutaraldehyde crosslinked polymers pose were prepared using the established solvent casting method as de-
risk of post implantation depolymerisation and further release of mono- scribed by Dong et al. [12]. (Fig. 1) Individual mixed solutions were
mers. Glutaraldehyde has further been reported to accelerate the calci- then poured in plastic containers and kept in an incubator at 45 C for
cation of the implanted prostheses [10]. On the other hand, glyoxal has 34 days. The lms were carefully peeled off from the plastic plates
been shown to be cytocompatible and support viability of the cells [11]. for further evaluations. The quantitative details of all the ingredients
In the experiment reported here we have developed a tripolymeric and are provided in Table 1.
a bipolymeric lm having natural and semi-natural biopolymers using
glyoxal as the crosslinker and reported a few physicochemical and me-
chanical properties of these biopolymeric lms crosslinked with glyoxal 2.4. Thickness measurement of lms
compared to those without any crosslinker.
The thickness of the lms was measured using a manual vernier cal-
2. Materials and method iper. The thickness was measured at different portions and an average
was calculated. The least count of the vernier caliper was 0.1 mm.
2.1. Materials

Chitosan [GRM9358] and CMC [GRM329] were procured from 2.5. Surface pH measurement
HiMedia laboratories India. Sodium Alginate [CAS 9005383] and PVA
[CAS 9002895] were purchased from Loba Chemicals India while The surface pH of the polymeric lms is an important consideration
Glyoxal [92245072735] was purchased from SRL Chemicals. Glycerol for drug delivery at particular site. An acidic or alkaline pH of the lm
[Merck; AK7AF57508] was used as the plasticizer. Acetic acid [Merck; surface could be erosive for the buccal mucosa or epidermal surface.
AB8A580103] was used as the solvent content for dissolving chitosan. For transbuccal and transdermal drug delivery, the surface pH should
be between 6.5 and 7.4, as per the physiological considerations. The
2.2. Preparation of polymeric solutions lms were washed with NaOH followed by distilled water to bring the
pH to physiological level in accordance with buccal mucosa and epider-
Chitosan, being soluble in acidic solution, was dissolved in 2% acetic mal surface. The pH was measured using a pocket-size pH meter by
acid (v/v) to form 1% (w/v) chitosan solution. CMC (1% w/v), Alginate Eutech Instruments. The experiment was replicated to cross-check
(1% w/v) and PVA (5% w/v) were dissolved in double distilled water. each value obtained
All the solutions were kept for 4 h stirring on magnetic stirrer. The solu-
tions were kept undisturbed overnight to get degassed.

2.3. Preparation of blended lms


Table 1
Quantitative details of the ingredients in polymeric lms.
The blended lms were formed in two combinations; Bipolymeric
(Chitosan + PVA or ChPVA lms) and Tripolymeric (CMC + Film formulation CMC Alginate Chitosan PVA Glycerol Glyoxal
Alginate + PVA or CAP lms). A total of 4 lms were developed, 2 F1 (CAP) 10 ml 10 ml 10 ml 3 ml 1.5 ml
with crosslinker and 2 without crosslinker. Initially, CMC (1% w/v), Algi- F2 (CAP + G) 10 ml 10 ml 10 ml 3 ml 1.5 ml
nate (1% w/v), chitosan (1% w/v in 2% acetic acid) and PVA (5% w/v) F3 (ChPVA) 15 ml 15 ml 3 ml 1.5 ml
F4 (ChPVA + G) 15 ml 15 ml 3 ml 1.5 ml
were prepared as the stock polymer solution. Glycerol (10% v/v) was
A. Kumar, A. Kumar / Materials Science and Engineering C 73 (2017) 333339 335

purpose, 50 ml agar solution (2% w/v) was prepared and the constitu-
ents of simulated salivary uid, as described by Koland et al., were
added separately in corresponding concentration [13]. The simulated
salivary uid constituents used were disodium phosphate, potassium
di-hydrogen phosphate and sodium chloride. The nal agar solution
was then poured in petri-dishes and left to get solidied. The petri-
dishes were kept for further use. A 2 cm2 piece was cut from all the
four lms. Each piece was kept on each agar plate prepared. The plates
were kept in incubator at 37 C. Each piece was weighed (W0) before
being placed on agar plate. Then weights were calculated for each
piece at an interval of 1 h for 4 h. The change in weight was measured
as swelling percentage and it was calculated as:

Wt W0 X100
W0

where Wt is the weight after each swelling interval, W0 is the initial


Fig. 2. Swelling index of the prepared polymeric lms. weight of the piece.

2.6. Swelling study 2.7. Surface characterization by SEM

All the four lms were subjected to swelling study under simulated The lms were subjected to surface morphology characterization
condition which revealed the water absorbing capacity. For this using Scanning Electron Microscope (SEM) [ZEISS EVO Series Scanning

Fig. 3. Camera photographs of the prepared lms.

Fig. 4. SEM images showing the surface morphology of the biopolymeric lms.
336 A. Kumar, A. Kumar / Materials Science and Engineering C 73 (2017) 333339

Fig. 5. EDX conrms the presence of sodium in the blend containing alginate.

Electron Microscope, Model EVO 18] available at Department of Metal- 2.9. Tensile strength measurement
lurgical Engineering, NIT Raipur, Chhattisgarh, India. The samples were
coated with gold before analysis using the Quorum-SC7620 Sputter Tensile strength and the stress-strain curve were obtained for all the
Coater. The samples were subjected to EDX analysis for conrmation four lms using Instron 3369, Bioplus, USA. The dimension of each sam-
of possible ingredients. ple was kept same at 20 mm length and 5 mm width. The operation was
done at a speed of 50 mm/min.

2.8. XRD analysis 2.10. Hardness testing

The lms were subjected to X-ray diffraction analysis [PANalytical Hardness testing on the biopolymeric lms was performed using
3 kW X'pert Powder Multifunctional] available with Department of Durometer Hardness (Shore A) Method. This method describes the in-
Metallurgical Engineering, NIT Raipur, Chhattisgarh, India. XRD analysis dentation hardness and is applied for materials such as thermoplastic
shows the interaction among the polymeric ingredients based on differ- elastomers, rubbers, cellular materials and polymeric materials. A du-
ence in the peaks of individual polymers and mixed polymers in lms. rometer (shore A type) instrument was used for this purpose. The in-
The analysis was done at 2 scanning range of 0100 at 45 kV. strument was pressed at 6 different surface positions of the polymeric

Fig. 6. XRD graphical representation of the interaction among biopolymers.


A. Kumar, A. Kumar / Materials Science and Engineering C 73 (2017) 333339 337

Table 2 3. Results
Various data of the polymeric lms obtained from tensile testing.

Film Length Width Maximum Tensile Extension Elongation 3.1. Thickness of the lm
composition (mm) (mm) load (N) strength at break percentage
(MPa) (mm) (%) The average thickness of the lm was obtained to be 0.878 mm for
F1 (CAP) 20 5 3.19 1.64 43.71 218.5 lm F1, 0.759 mm for lm F2, 0.846 mm for lm F3 and 0.784 mm for
F2 (CAP + G) 20 5 0.91 0.46 49.68 248.4 lm F4 respectively. The average thickness of the four lms was obtain-
F3 (ChPVA) 20 5 0.42 0.22 3.14 15.7
ed to be 0.816 mm.
F4 20 5 0.92 0.74 6.82 34.1
(ChPVA + G)

3.2. Surface pH

The surface pH of lms was found to be 6.4, 6.2, 6.4, and 6.8 for lms
F1, F2, F3 and F4 respectively. This pH is acceptable for the lms to be
used for transbuccal or transdermal drug delivery, since the surface of
buccal mucosa and epidermis favors the pH in this particular range.

3.3. Swelling study

The swelling study revealed that the effect of crosslinking with


glyoxal was negligible on the swelling index of CAP lms as they
demonstrated a slight decrease in weight from the 2nd hour of swell-
ing, even when a good swelling was observed after 1st hour. The sim-
ple non-crosslinked CAP lm displayed a subsequent decrease in
weight from the 2nd hour of swelling. The glyoxal crosslinked
ChPVA lm displayed better swelling than its counterpart non-
Fig. 7. Combined graphical representation of tensile strength, elongation at break and crosslinked ChPVA. The simple ChPVA displayed uniform swelling
percent elongation of the lms.
for the initial 2 h and a steep uniform increase for next 2 h while
the crosslinked ChPVA displayed a continuous swelling for 1st and
lms giving a hardness value and a mean value was calculated. All the 2nd hour but the swelling increased and remained stable for next
testing were in accordance with ASTM D2240 and IS 13360 (part 5/sec- 2 h. The swelling index graph is represented in Fig. 2. The swelling
tion 11: 1992) protocols. The hardness test was performed at Central In- of blended polymeric lms is necessary for the loaded drugs to get
stitute of Plastics Engineering and Technology (CIPET), Raipur. released.

Fig. 8. Load vs. strain graph of the lms.


338 A. Kumar, A. Kumar / Materials Science and Engineering C 73 (2017) 333339

Fig. 9. Load vs. extension graph of the lms.

3.4. SEM analysis a blunt peak at 2 angle of 10 and a sharp short peak at 2 angle of 21
while for ChPVAG (F4) lm, a single sharp peak was observed at 2
The surface morphology analysis using SEM revealed a porous rough angle of 20 while no other peak was observed in the graph. These XRD
surface of the non-crosslinked CAP lms while the crosslinked CAP lms images revealed a signicant interaction of polymers in the composite
displayed a much smoother surface. Similar to the morphological lms. The corresponding XRD peaks have been shown in Fig. 6.
change in CAP lms, glyoxal crosslinking resulted signicant change in
surface morphology for ChPVA lms too. The camera images of the 3.6. Tensile strength and stress-strain curve
lms are shown in Fig. 3. The respective SEM images of the lms are
shown in Fig. 4. Since the lms were developed by solvent casting meth- The tensile strength indicates the ability of the sample to resist the
od, a high porosity was indeed not expected. But a microscopic surface tension. The parameters obtained from tensile testing have been
analysis was performed to check the effect of glyoxal as crosslinker. Fur- shown in Table 2. The tensile strength (MPa) for F1, F2, F3 and F4 was
ther, the conrmation of ingredients was done using Energy Disruptive obtained to be 1.64, 0.46, 0.22 and 0.74 respectively. The maximum
X-ray imaging (EDX). Fig. 5 shows the presence of sodium alginate in load (in Newton or N) for F1, F2, F3 and F4 was obtained as 3.19, 0.91,
the CAP and CAPG lms. 0.42 and 0.92 respectively. Upon calculating percentage elongation
from extension at break data [14], we report here that the percent elon-
gation of non-crosslinked CAP lms (F1) was 218% while for crosslinked
3.5. XRD interaction analysis CAP lms (F2) was 248%. For non-crosslinked ChPVA lms (F3) we ob-
tained a percentage elongation of 15% while it was more than double at
The XRD analysis of the individual polymeric ingredients and the 34% for glyoxal crosslinked ChPVA lms (F4). Since non-crosslinked CAP
composite lms revealed a clear and signicant interaction among the
individual polymers in the lms. Alginate showed two sharp at 2
angle value of 15 and 23, chitosan displayed its sharp intensities at 2
angle of 10 and 20, CMC displayed a sharp peak at 2 angle of 20
while PVA displayed a short peak at 2 angle of 12, a sharp peak at 2
angle of 20 and another short peak at 2 angle of 40. For the rst lm
CAP (F1), blunt plateau intensity was observed at 2 angle of 13 while
a sharp peak at 2 angle of 21. The sharp peak of alginate at 2 value
15 disappeared while the sharp peak at 23 displayed a mild left hand
deected. For CAPG (F2) lm, we observed a blunt peak at 2 value of
10 while a relatively sharp peak at 22. For ChPVA (F3) lm, we observed

Table 3
Mean durometer hardness value of the biopolymeric lms.

S. no. Film composition Hardness value (shore A)

1. CAP (F1) 19
2. CAP + G (F2) 16.14
3. ChPVA (F3) 56.5
4. ChPVA + G (F4) 53.8
Fig. 10. Mean durometer hardness plot of the polymeric lms.
A. Kumar, A. Kumar / Materials Science and Engineering C 73 (2017) 333339 339

lms already had a signicant percent elongation, the crosslinking did improve the exibility of CMC, alginate, and PVA (CAP) containing
not add much to this property. On the other hand a signicant increase lms, it did signicantly improve the property for chitosan and PVA
in percent elongation (more than double) demonstrates a good (ChPVA) constituted the lm. The swelling index of CAP lms remained
crosslinking between chitosan and PVA. These results indicate that almost similar in crosslinked and non-crosslinked lms while it in-
while glyoxal crosslinking decreased the tensile strength signicantly creased signicantly in ChPVA lms. The surface morphology analysis
of the lms constituting CMC, Alginate and PVA, it worked well to showed that the crosslinked lms have the ne surface as compared
crosslink the lms made of chitosan and PVA and increased the corre- to the rough indication in non-crosslinked lms. The durometer hard-
sponding tensile strength. The collective graphical representation of ness testing further revealed that adding glyoxal as crosslinker reduced
tensile strength, extension at break and elongation percent has been the hardness of the compositions. Further, even when the comparative
shown in Fig. 7. The corresponding tensile strength graph and stress swelling index has not been good in our study for CAP lms, they can
vs. strain graph obtained through the instrument operation has been still be propagated for transbuccal and transdermal drug delivery appli-
shown in Fig. 8 and Fig. 9. cation. Further in vitro and in vivo studies are required using different
polymeric combinations which can focus more on the crosslinking ap-
3.7. Hardness testing plication of glyoxal in the biomedical eld. These kinds of composite
multi-polymeric lms could be applied for drug delivery, and wound
The durometer hardness testing revealed that the CMC and alginate healing coverings.
based polymeric lms were signicantly softer (less hard) as compared
to the chitosan based lms. Further, upon crosslinking with glyoxal, the Conict of interest
mean hardness of the lms decreased. As reported in section 3.6 previ-
ously, the crosslinking effect of glyoxal on tensile strength was observed Authors declare no conict of interest.
as greater percentage of elongation in the crosslinked lms and that also
favor our hardness test since glyoxal crosslinked lms became less hard
as compared to the non-crosslinked lms. The mean values of hardness Acknowledgement
for all the four lms have been presented in Table 3 and a comparative
hardness has been plotted in Fig. 10. Authors are highly thankful and acknowledge the kind support and
facilities provided by Department of Biotechnology, National Institute
4. Conclusion of Technology Raipur, and CIPET Raipur, Chhattisgarh, India.

Biopolymeric lms have great acceptability for various biomedical References


applications like drug delivery (transdermal or transbuccal route), tis- [1] K.V. De Velde, P. Kiekens, Biopolymers: overview of several properties and conse-
sue engineering etc. Natural and semi-synthetic (or semi-natural) bio- quences on their applications, Polym. Test. 21 (2002) 433442.
polymers are even better as they have least adverse effects on the [2] A.S. Hoffman, Hydrogels for biomedical applications, Adv. Drug Deliv. Rev. 54 (2002)
312.
physiological system. Chitosan, alginate, and carboxymethyl cellulose [3] A. Kumar, A. Vimal, A. Kumar, Why chitosan? From properties to perspective of mu-
have been the most applied polysaccharide biopolymers for the various cosal drug delivery, Int. J. Biol. Macromol. 91 (2016) 615622.
biomedical applications. In this article, we have prepared lms using [4] V.V. Khutoryanskiy, Advances in mucoadhesion and mucoadhesive polymers,
Macromol. Biosci. 11 (2011) 748764.
different combinations of chitosan, alginate, CMC and PVA. The empha- [5] R. Shaikh, T.R. Raj Singh, M.J. Garland, A.D. Woolfson, R.F. Donnelly, Mucoadhesive
sis lies here in glyoxal which is a widely used crosslinker in the textile drug delivery systems, J. Pharm. Bioallied Sci. 3 (1) (2011) 89100.
industry, plastic industry, and in leather tanning. Glyoxal is the smallest [6] A. Amit, S. Dwivedi, Ajazuddin, T.K. Giri, S. Saraf, S. Saraf, et al., Approaches for break-
ing the barriers of drug permeation through transdermal drug delivery, J. Control.
dialdehyde and has a great crosslinking application for polysaccharide
Release 116 (2012) 2640.
polymers. It forms hemiacetal bonds with the hydroxyl groups present [7] K. Azuma, R. Izumi, T. Osaki, S. Ifuku, M. Morimoto, et al., Chitin, chitosan, and its de-
in the polysaccharides [15]. rivatives for wound healing: old and new materials, J. Funct. Biomater. 6 (2015)
Glutaraldehyde (one of the widely used crosslinker) has been re- 104142.
[8] T. Dai, M. Tanaka, Y.Y. Huang, M.R. Hamblin, Chitosan preparations for wounds and
ported to be relatively toxic in nature especially for the respiratory burns: antimicrobial and wound-healing effects, Expert Rev. Anti-Infect. Ther. 9 (7)
tract. It also has reported irritation on eyes and skin with reported der- (2011) 857879.
matitis and rhinitis. Occupational asthma has also been reported for re- [9] C. Paczyski, J. Walusiak, U. Ruta, P. Grski, Occupational asthma and rhinitis due to
glutaraldehyde: changes in nasal lavage uid after specic inhalatory challenge test,
spiratory technologists who use glutaraldehyde for sterilization of Allergy 56 (12) (2001) 11861191.
endoscopes [9]. In another study related to cellular growth over bio- [10] C.M. Vaz, L.A. de Graaf, R.L. Reis, A.M. Cunha, In vitro degradation behaviour of bio-
scaffold of collagen and PVA, it was found that glutaraldehyde induces degradable soy plastics: effects of crosslinking with glyoxal and thermal treatment,
Polym. Degrad. Stab. 81 (2003) 6574.
apoptosis in the cells and thus demonstrated poor cell growth over glu- [11] L. Wang, J.P. Stegemann, Glyoxal crosslinking of cell-seeded chitosan/collagen
taraldehyde crosslinked scaffolds. The mechanism was attributed to the hydrogels for bone regeneration, Acta Biomater. 7 (6) (2011) 24102417.
glutaraldehyde on the surface of the scaffolds tested in the study [16]. [12] Z. Dong, Q. Wang, D.Y. Alginate, Gelatin blend lms and their properties for drug
controlled release, J. Membr. Sci. 280 (2006) 3744.
Thus, biopolymeric lms crosslinked with glutaraldehyde could have [13] M. Koland, R.N. Charyulu, K. Vijayanarayana, P. Prabhu, In vitro and in vivo evalua-
potential adverse effects towards their use as transdermal and tion of chitosan buccal lms of ondansetron hydrochloride, J. Pharm. Investig. 1 (3)
transbuccal applications. Glyoxal has been used as an alternative (2011) 164171.
[14] https://www.ipc.org/TM/2.4.18.3.pdf; accessed on July 20, 2016.
dialdehyde crosslinker in various biomedical studies and applications
[15] K.S. Mikkonen, M.I. Heikkila, S.M. Willfor, M. Tenkanen, Films from glyoxal-
and is considered safe [10,11]. From this study we can say that crosslinked spruce galactoglucomannans plasticized with sorbitol, Int. J. Pol. Sci.
crosslinking with glyoxal does improve the physico-chemical and me- (2012) 18, 482810. .
chanical properties of chitosan-based composite polymers much signif- [16] J.E. Gough, C.A. Scotchford, S. Downes, Cytotoxicity of glutaraldehyde crosslinked
collagen/poly(vinyl alcohol) lms is by the mechanism of apoptosis, J. Biomed.
icantly than carboxymethylcellulose and alginate based polymers. The Mater. Res. 61 (1) (2002) 121130.
results indicate that on one hand where glyoxal does not signicantly

Anda mungkin juga menyukai