Biomaterials Research (2012) 16(1) : 25-31
25
Biomaterials
Research
C The Korean Society for Biomaterials
Fabrication of Hyaluronic Acid-Loaded Polyvinyl Alcohol
HydrogelPatch and Evaluation of Wound Healing Efficacy
1
1
1
2
2
3
1*
Bo Ra Nam1, Se Heang Oh1, Sang June Kim1, Dong Young
Kang2,Seungho Lee2, Kyu Sang Song3, and Jin Ho Lee1*
1 , 2 , 3 1Department of Advanced Materials, Hannam University,
Daejeon 305-811, Korea2Department of Chemistry, Hannam University,
Daejeon 305-811, Korea3Department of Pathology, School of Medicine,
Chungnam National University, Daejeon 301-131, Korea(Received
January 5, 2012/Acccepted February 2, 2012)
In this study, we fabricated hyaluronic acid (HA)-loaded
polyvinyl alcohol (PVA) hydrogel patches as a wound dressingby a
simple freezing/thawing method to estimate the effect of HA
addition and its molecular weight (MW) during thewound healing. The
molecular weight of HA was controlled by a -ray irradiation [HA
with high MW (HHA, ~4,800kDa) & HA with low MW (LHA, ~52 kDa)].
The prepared PVA/HA hydrogel patches were flexible and elastic,
andhad a high swelling property in water. The wound healing
behavior of the PVA/HA hydrogel patches were comparedwith those of
PVA hydrogel patch as well as gauze (control) using SD rats as a
wound model. It was observed thatthe PVA/HA hydrogel patches had
faster wound healing behavior than gauze and PVA patch groups,
regardless ofmolecular weight of HA. It is probably owing to the
synergistic effect of PVA which allows the fast absorption of
exu-date from wound and provides wet environment for wound, and HA
which provides the stimulation of cell pro-liferation for wound
healing. It was also observed that the PVA/HHA hydrogel patch
showed more effective woundhealing behavior than the PVA/LHA one,
however, the difference is not significant. This result suggests
that the PVA/HA hydrogel patch can be a good candidate as a
dressing material for effective wound healing.
Key words: Wound dressing, polyvinyl alcohol, hyaluronic acid,
hydrogel
/
,
, ,
.1)
(infection) (dehydration) ,
(wound dressing)
.2,3)
,
/
,
, , ,
, , , , ,
, .4)
(polyurethane),
(polyethylene), (polycaprolactone),
(polyacrylonitrile), (silicone rubber)
(alginate), (chitosan), (gelatin),
(collagen)
,5) 3
,
.6)
[(polyhydroxyethyl methacrylate), p(HEMA)],
(polyethylene oxide, PEO), (polyvinyl
alcohol, PVA) (polyacrylamide, PAA)
,6)
(biocompatibility), (non-toxicity), (non-carcinoge-
nicity), (high mechanical properties),
(high water content) (easy processing)
7) .
*: [email protected]
26
Vol. 16, No. 1
(extracellular matrix, ECM)
glycosaminoglycan(GAG; or mucopolysaccharide)
(hyaluronic acid, HA) -D-glucuronic acid
-D-N-acetylglucosamine
1,000 Da 10,000,000 Da
, .8) 1934
Meyer Palmer (bovine vitreous body)
,9) (high molec-
ular weight)
, , ,
, /
.8) , ,
(differentiation) (proliferation) ,
,10-12)
, ,
.13-15)
(hemostasis),
(imflammation), (granulation tissue forma-
tion), (re-epithelization) (re-modeling)
.16-19)
,
,
(network)
,
keratinocyte
.20)
,
.21-23) ,
.24,25)
/
, , (swelling ratio)
/
.
(MW 89,000~98,000 g/mol; hydrolysis ~99%;
Sigma-Aldrich, USA) ,
//
(HHA; MW, ~4,800 kDa; Shiseido, Japan)
.
(LHA; MW, ~52 kDA)
.
,
.
(JS-10000, Soyagreentec, Korea) 100 kGy
.
multi-angle laser light scat-
tering(MALLS) (field-flow
fractionation method) .
, [ (DMX-2003T, SMK
Inc., Japan), Peristaltic (M312, GILSON, France)],
(Rheodyne 7725, Grace, USA), Field-flow fractionator
(F-1000, Fractionation Research Inc., USA), (M720,
YOUNG-LIN, Korea), .
Field-flow fractionator data
(Fractionation Research Inc.) .
needle valve , (buret, PYREX, USA)
.
(deionized water)
90oC 5 wt% ,
1 wt%
/ .
38 mm (polystyrene, PS)
2 mL 76oC 18 /
6
(freezing/thawing method),
/
(diameter ~38 mm, thickness ~2 mm).
,
. ethylene oxide(EO) gas
.
(ASTM
27
Biomaterials Research 2012
D 638 type V) dog-bone
, ultimate tensile test machine(UTM; AG-5000G,
Shimadzu, Japan) crosshead 10 mm/min
.
24
.
, ,
(diameter ~38 mm, thickness ~2 mm)
(Wd), (~40 mL) 37oC
incubator (1, 2, 3, 5, 7, 12, 24 )
.
(Ws).
(swelling ratio) .
Swelling ratio(%) = (Ws Wd)/Wd 100
(in vivo)
, / ,
/
, (Sprague-Dawley(SD) rat,
200~300 g) .
Zoletil 50(Virbac Laboratories, France) 50 mg/kg xylazine
hydrochloride(Rumpun, Bayer, Korea) 5 mg/kg
. clipper
betadine 70% ,
2 cm
(1 defect/rat). EO gas
(diameter ~38 mm, thickness ~2 mm)
,
(Coban, 3M, USA) . ,
(3 3 cm2) (control)
. 3 7
, 2 .
,
(i-
solution, IMT Inc., Korea) ,
H
& E .
, MALLS
.
100 kGy
, Table 1
.
, 100 kGy 4,800 kDa 52
kDa 1/10 .
backbone chain
,
backbone chain
,
.26) ,
,
.
Figure 1 ( )
. 260 gf/mm2
(fracture strength) 6% (elongation)
, 120 gf/mm2 314%
. ,
. , ,
, / [PVA/HHA; ,
~200 gf/mm2, , ~4%(dry state); , ~20 gf/
mm2, , ~245%(swollen state)], /
[PVA/LHA; , ~110 gf/
mm2, , ~2%(dry state); , ~10 gf/mm2, ,
~140%(swollen state)]
.
( )
( )
,
. Kim 27) 7 gf/mm2
.
, ,
Figure 2 . ,
Table 1. Average molecular weights of -ray irradiated hyaluronic
acids
-ray irradiation dose (kGy) Molecular weight (kDa)
0 4,800
15 310
25 146
50 93
100 52
31
Biomaterials Research 2012
2011
.
1. D. O. Han, G. H. Kim, Y. B, Choi, I. S. Shim, and H. J.
Lee,Healing effects of astragali radix extracts on experimental
openwound in rat, Oriental Physiol. Pathol., 19, 92-97 (2005).
2. I. V. Yannas, J. F. Burke, Design of an artificial skin. I.
Basicdesign principles, J. Biomed. Mater. Res., 14, 65-81
(1980).
3. N. Dagalakis, J. Flink, P. Stasikelis, J. F. Burke, I. V.
Yannas, Designof an artificial skin. II. Control of chemical
composition, J.Biomed. Mater. Res., 14, 107-132 (1980).
4. M. Szycher and S. J. Lee, Modern wound dressings: a
systematicapproach to wound healing, J. Biomater. Appl., 7,
142-213(1992).
5. J. W. L. Davies, Synthetic materials for covering burn
wounds:progress towards perfection. Part I. Short term dressing
materials,Burns, 10, 94-103 (1983).
6. P. H. Corkhill, C. J. Hamilton, and B. J. Tighe, Synthetic
hydrogelsVI. Hydrogel composites as wound dressings and
implantmaterials, Biomaterials, 10, 3-10 (1989).
7. C. M. Hassan and N. A. Peppas, Structure and applications
ofpoly (vinyl alcohol) hydrogels produced by conventional
cross-linking or by freezing/thawing method, Adv. Polym. Sci., 153,
37-65 (2000).
8. Http://en.wikipedia.org/wiki/Hyaluronan9. I. S. Jo and S. K.
Min, The effect of hyaluronic acid on xenograft
in rat calvarial defect, J. Korean Assoc. Oral maxillofac.
surg., 28,205-215 (2002).
10. I. R. Hong and Y. J. Kim, Self-Aggregated nanoparticles of
lipoicacid conjugated hyaluronic acid, Polymer (Korea), 32,
561-565(2008).
11. K. T. Kim, Y. H. Kim, J. G. Kim, C. S. Han, S. H. Park, K.
H. Kim,Preparation of oligo hyaluronic acid by hydrolysis and
itsapplication as a cosmetic ingredient, J. Soc. Cosmet.
ScientistsKorea, 33, 189-196 (2007).
12. S. K. Hahn, E. J. Oh, H. Miyamoto, and T. Shimobouji,
Sustainedrelease formulation of erythropoietin using hyaluronic
acidhydrogels crosslinked by Michael addition, Int. J. Pharm.,
322,44-51 (2006).
13. D. D. Allison and K. J. Grande-Allen, Review hyaluronan:
Apowerful tissue engineering tool, Tissue Eng., 12,
2131-2140(2006).
14. S. J. Kim, C. K. Lee, Y. M. Lee, I. Y. Kim, and S. I. Kim,
Electrical/pH-sensitive swelling behavior of polyelectrolyte
hydrogelsprepared with hyaluronic acid-poly(vinyl alchol)
interpenetratingpolymer networks, React. Funct. Polym., 55, 291-298
(2003).
15. G. D. Prestwich, D. M. Marecak, J. F. Marecek, K. P.
Vercruysse,and M. R. Ziebell, Controlled chemical modification
ofhyaluronic acid: Synthesis, applications, and biodegradation
ofhydrazide derivates, J. Control Release 53, 93-103 (1998).
16. K. M. Choi, C. W. Lee, and M. Y. Lee, Effect of ore minerals
on
the healing of full-thickness skin injury model of rat, J.
Environ.Sci., 17, 809-816 (2008).
17. S. H. Qi, P. Liu, J. L. Xie JL, Y. B. Xu, C. N. Ke, X. S.
Liu, and T. Z.Li, Experimental study on repairing of nude mice skin
defectswith composite skin consisting of xenogeneic dermis
andepidermal stem cell and hair follicle dermal papilla cell,
Burns,34, 385-392 (2008).
18. A. J. Singer and R. A.F. Clark, Cutaneous wound healing,
NewEngl. J. Med., 341, 738-746 (1999).
19. S. O. Iseri, I. E. Gedik, C. Erzik, B. Uslu, S. Arbak, N.
Gedik, andB. C. Yegen, Oxytocin ameliorates skin damage and
oxidantgastric injury in rats with thermal trauma, Burns, 34,
361-369(2008).
20. W. Y. Chen and G. Abatangelo, Functions of hyaluronan
inwound repair, Wound Repair Regen., 7, 79-89 (1999).
21. Y. Matsumoto, K. Arai, H. Momose, and Y. Kuroyanagi,
Develop-ment of a wound dressing composed of a hyaluronic acid
spongecontaining arginine, J. Biomater. Sci. Polym. Ed., 20,
993-1004(2009).
22. F. Gao, Y. Liu, Y. He, C. Yang, Y. Wang, X. Shi, and G.
Wei,Hyaluronan oligosaccharide promote excisional wound
healingthrough enhanced angiogenesis, Matrix Biol., 29, 107-116
(2010).
23. C. J. Doillon and F. H. Silver, Collagen-based wound
dressing:Effects of hyaluronic acid and firponectin on wound
healing,Biomaterials, 7, 3-8 (1986).
24. S. Gotoh, J. Onaya, M. Abe, K. Miyazaki, A. Hamai, K.
Horie,and K. Tokuyasu, Effects of the molecular weight of
hyaluronicacid and its action mechanisms on experimental joint pain
inrat, Ann. Rheum. Dis., 52, 817-822 (1993).
25. J. Kim, Y. Park, G Tae, K. B. Lee, C. M. Hwang, S. J. Hwang,
I. S.Kim, I. Noh, and K. Sun, Characterization of
low-molecular-weight hyaluronic acid-based hydrogel and
differential stem sellresponses in the hydrogel microenvironments,
J. Biomed. Mater.Res., 88A, 967-975 (2009).
26. T. Kume and M. Takehisa, Effect of gamma -irradiation
onsodium alginate and carrageenan powder, Agric. Biol. Chem.,47,
889-890 (1983).
27. W. I. Kim, C. J. Kim, D. Y. Kim, O. K. Kwon, and O. H.
Kwon,Fabrication and characterization of polyurethane foam forwound
dressing, Polymer (Korea), 34, 442-449 (2010).
28. S. C. Park, S. H. Oh, and J. H. Lee,
Alginate/aloe-containingpolyvinyl alcohol hydrogel patch as a wound
dressing, Biomater.Res., 13, 138-143 (2009).
29. S. J. Kim, S. G. Yoon, Y. M. Lee, H. C. Kim, and S. I. Kim,
Elec-trical behavior of polymer hydrogel composed of
poly(vinylalcohol)-hyaluronic acid in solution, Biosens.
Bioelectron, 19, 531-536 (2004).
30. V. A. Luprano, P. A. Ramires, G. Montagna, and E. Milella,
Non-destructive characterization of hydrogels, J. Mater. Sci.
Mater.Med., 8, 175-178 (1997).
31. M. Kobayashi, J. Toquchida, and M. Oka, Development
ofpolyvinyl alcohol-hydrogel (PVA-H) shieles with a high
watercontent for tendon injury repair, J. Hand Surg. Br., 26,
436-440(2001).
32. G. S. diZerega, Contemporary adhesion prevention, Fertil.
Steril.,61, 219-235 (1994).
