Journal of Controlled Release 64 (2000) 313 www.elsevier.

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Biodegradable amphiphilic multiblock copolymers and their implications for biomedical applications
You Han Bae*, Kang Moo Huh, Youngso Kim, Keun-Hong Park
Department of Materials Science and Engineering, Kwangju Institute of Science and Technology, 572 Sangam-dong, Kwangsan-ku, Kwangju 506 -712, South Korea Received 1 April 1998; accepted 13 April 1999

Abstract Alternating multiblock copolymers composed of short blocks of poly(ethylene oxide) (PEO) and poly(-caprolactone) (PCL) or poly( L-lactic acid) (PLLA) were synthesized by a coupling reaction. The block copolymers of relatively high molecular weights (Mn . 20,000) formed a physically crosslinked thermoplastic network, while low molecular weight polymers were water-soluble. The block copolymers demonstrated solubility in a variety of solvents including acetone, tetrahydrofuran, methylene chloride, dioxane, water / acetone mixtures, and water / ethanol mixtures. The degree of swelling, optical transparency, and mechanical property of the lms, prepared by a solvent casting method, were affected by the nature of the hydrophobic block used, polymer composition, temperature, and thermal history. The crystalline melting temperatures of PCL and PLLA in the block copolymers were signicantly lowered due to the chemical structure of difunctional PCL and PLLA, and partial phase mixing with PEO segments. The properties of the block copolymers may be useful for biomedical applications as well as controlled drug release formulations. When PEO / PLLA multiblock copolymers were applied as a wound healing material loaded with basic broblast growth factor (bFGF), the feasibility study showed improved wound healing when compared to controls of no treatment and the same wound covering without bFGF, indicating that a certain degree of the bioactivity of bFGF is preserved. 2000 Elsevier Science B.V. All rights reserved.
Keywords: Poly(ethylene oxide); Poly(-caprolactone); Poly( L-lactic acid); Alternating multiblock copolymers; Physical crosslinking; Thermoplastic biodegradable hydrogel; Wound healing

1. Introduction Hydrogels, which absorb a signicant amount of water but retain their three-dimensional structure in the swollen state [13], are three dimensional hydrophilic polymer networks formed by chemical or physical crosslinking. If the polymer is crosslinked
*Corresponding author. Tel.: 182-62-970-2305; fax: 182-62970-2304. E-mail address: yhbae@matla.kjist.ac.kr (Y.H. Bae)

covalently, it is called a chemical gel [3]. Chemical gels can be prepared by the crosslinking of linear polymers which involves organic reactions between pendant functional groups of linear polymers, or by simultaneous polymerization using multifunctional monomers. These are generally brittle and difcult to process. Physical gels maintain their ordered structure by non-covalent cohesion interactions, such as hydrophobic interaction, stereocomplex formation, ionic complexation and crystallinity [46]. The resultant

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physical associations act as physical junction domains, which are not crosslinking points but chain segments involving 15 or more residues [7]. Although non-covalent associations are reversible and weaker than chemical crosslinking, they allow solvent casting and thermal processing, and the resulting polymers often show elastic or viscoelastic properties. However, most known physical gels lack controlled degradability which limits their use in the body, especially for delivery applications. Although biodegradable polymers such as polylactide, polyglycolide, and their copolymers are often employed as delivery carriers for various drugs, they do not full all properties required for a wide range of delivery purposes. Biodegradable physical hydrogels may offer an alternative material of choice in designing drug delivery systems as well as other biomedical applications. In this research, hydrophilic / hydrophobic multiblock polymers were prepared through coupling reactions between two short blocks without using a chain extender: biocompatible poly(ethylene oxide) (PEO) with degradable poly(-caprolactone) (PCL) or poly( L-lactide) (PLLA). Most block copolymers containing PEO and PCL or PLLA are related to diand triblock copolymers, obtained by ring opening polymerization of -caprolactone or lactide at the end of PEO [8]. Such conventional block copolymers have a limitation in increasing molecular weight without using long PEO blocks, which are highly likely to accumulate in the body after degradation, for targeted amphiphilicity. However, multiblock copolymers constructed with relatively short blocks can be used to produce high molecular weight polymers with considerable mechanical strength [9]. Another interest in the multiblock copolymer lies in obtaining many possible combinations of short blocks. In this investigation, the copolymers were obtained by esterication of dicarboxylated PEO and dihydroxy PCL for PEO / PCL multiblocks, and dihydroxy PEO and dicarboxylated PLLA for PEO / PLLA multiblocks. This paper reports the unique properties of the multiblock copolymers in temperature-dependent swelling, optical transmittance, and mechanical strength. These qualities make them useful for controlled and / or triggered drug delivery and other biomedical applications.

Preliminary results of a trial using PEO / PLLA multiblock copolymers loaded with basic broblast growth factor as a wound covering material are demonstrated. Basic broblast growth factor (bFGF) has desirable wound healing properties, being a potent promoter of angiogenesis [10,11] and a mitogen and chemoattractant for endothelial cells and broblasts [12]. bFGF is also a strong stimulator of collagenase and plasminogen activator production [13], initiating breakdown of the extracellular matrix and thereby facilitating the migration of cells through the matrix to the wound sites. However, one of the most challenging problems in the delivery of therapeutic polypeptides or protein is preservation of their stability in the delivery system. Although bFGF is known to be potent and benecial in stimulating cell growth and the wound healing process, pathophysiological studies are hampered by its instability [14]. For instance, when bFGF was released in a sustained fashion from a polymeric delivery system, 99% of its mitogenic activity was lost [15]. Thus we briey evaluated a PEO / PLLA multiblock hydrogel formulation containing bFGF for its effectiveness in preserving the healing properties of the substance.

2. Experimental

2.1. Materials
1,4-Dioxane and methylene chloride were dried over sodium and calcium, respectively, and then distilled. N,N-Dimethyl aminopyridine (DMAP), dicyclohexyl carbodiimide (DCC), dihydroxy-terminated poly(-caprolactone) (PCL-(OH) 2 , nominal Mw 1250) and poly(ethylene oxide) (PEO, nominal Mw s 1000 and 1500, 2000, and 3500) were purchased from Aldrich Chemical Co. (Milwaukee, WI). PCL(OH) 2 (nominal Mw 2000) and triethylamine (TEA) were received from Acros (Geel, Belgium) and succinic anhydride was purchased from Fluka AG (Switzerland). It was found by 1 H-NMR analysis that commercial PCL diols contain incorporated di(ethylene glycol) initiator residue originating from production. To remove residual moisture, PEO and PCL were dried at 708C under reduced pressure for 12 h prior to use. L-Lactic acid (85% aqueous

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solution, Aldrich) and succinic acid (Sigma) were used as received without further purication. Recombinant human basic broblast growth factor (bFGF) was obtained from Gibco (Grand Island, NY). All other chemicals and solvents were used as received.

2.2. Synthesis of PEO /PCL multiblock copolymers

The synthesis of PEO / PCL multiblock copolymers is presented in Scheme 1. Dicarboxylated PEO was prepared according to Zalipsky et al. [16]. Briey, dried PEO (Mw 1000, 6 mmol) was dissolved in 1,4-dioxane (30 ml). Succinic anhydride (14 mmol), DMAP (12 mmol) and triethylamine (12 mmol) were added into the solution and stirred for 24 h at room temperature under anhydrous conditions. The dioxane was evaporated in vacuo. The residue was dissolved in CCl 4 and the solid was ltered off. The ltrate was precipitated in diethyl ether, ltered, and dried in vacuo. The derivatization of PEO 1500 was performed by the same method. For the synthesis of a PEO / PCL (1000 / 1250) multiblock copolymer, dicarboxylated PEO 1000 (Mw 1322; 5.0 g, 3.8 mmol) and equimolar PCL (Mw 1250) were dissolved in methylene chloride (25 ml). DMAP (0.9 mmol) and DCC (9 mmol) were added

Scheme 2. Synthetic route for dicarboxylated PLLA

and stirred overnight at room temperature under dried nitrogen. Precipitated dicyclohexylurea (DCU) was removed by ltration. The ltrate was precipitated in cold diethyl ether (48C). The product was collected by ltration and dried in vacuo. The same procedure was used for other PEO / PCL multiblock copolymers.

2.3. PEO /PLLA multiblock copolymers

Dicarboxylated PLLA was prepared according to Scheme 2. L-Lactic acid in aqueous solution (100 g) was placed in a two-neck ask with a predetermined amount of succinic acid to obtain a desired molecular weight of PLLA. Dried nitrogen was continuously introduced in the reaction ask, and the temperature was gradually raised up to 1608C in a thermostat oil bath. After 24 h, the nitrogen supply was stopped

Scheme 1. Synthetic route for PEO / PCL multiblock copolymer.

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and the reaction ask kept at 1902008C in vacuo for 4 days. After cooling the product was dissolved in acetone and then precipitated in distilled water. The precipitate was dried in a vacuum oven over 2 days. PEO / PLLA multiblock copolymers were synthesized by a method similar to Scheme 1. Briey, PEO (5 mmol) and equimolar dicarboxylated PLLA were dissolved in 70 ml anhydrous methylene chloride with DMAP (1.3 mmol) and DCC (13 mmol). The coupling reaction was conducted in a dry nitrogen environment at room temperature while stirring for 12 h. After ltering DCU, the ltrate was concentrated under reduced pressure and then poured into an excess amount of cold diethyl ether with vigorous stirring. The precipitate was re-dissolved in methylene chloride and precipitated in a mixture of diethyl ether and methanol (9 / 1 v / v) for purication. The puried product was dried in a vacuum oven for 24 h.

using a tensile tester (Instron, Model 5567). The mechanical tests for lms (0.60.8 mm in thickness) of a dog-bone type according to ASTM D638M were performed in dried condition and hydrated for 24 h in distilled water at room temperature. Mechanical tests of the swollen samples after they had been heated above the PCL crystalline melting temperature were also undertaken. For this experiment, samples equilibrated in water at room temperature were placed in a sealed bottle without additional water and heated to 608C for 20 min and then cooled to room temperature. At least three separate tensile specimens were used for each polymer. Youngs moduli were obtained at room temperature at a crosshead speed of 20 mm / min.

2.5. Swelling
The polymer lms were prepared by a solvent casting method. A 10 wt.% polymer solution in methylene chloride was poured on a casting mold (at Teon plate), air-dried for 24 h, and vacuumdried until a constant weight was reached. The dried lm (|0.2 mm in thickness) was cut into squares with 1 cm width. The samples were immersed into a pH 7 PBS solution at desired temperatures. The weights of the swollen samples were monitored at specic time intervals after removing excess water on the surface by tapping with a lter paper. The weight swelling ratio was dened as Ww /Wp , where Ww 5 Whydrogel 2 Wp and Whydrogel and Wp are the weights in swollen and dried samples, respectively. The changes in swelling between two xed temperatures were obtained by alternately placing the samples in two thermostat water baths.

2.4. Polymer characterization

IR spectra were obtained by FT-IR (Perkin-Elmer 200 series). 1 H-NMR spectra were recorded on a JEOL JNM-LA 300 WB (300 MHz, TMS as an inert standard). The molecular weights and molecular weight distributions of the polymers were estimated by size exclusion chromatography on Waters StyragelE HR2, HR4 and HR5 column (in series) in THF at a ow rate 1 ml / min. Transmittance of polymer lms were measured using a Varian CARY 1E UV-VIS spectrophotometer equipped with a Varian CARY temperature controller. The cast lm was cut to the desired shape and attached to inside the wall of a styrene UV/ VIS spectrophotometer cuvette which was then lled with pH 7 phosphate buffered saline (PBS) solution. The transmittance of the swollen hydrogel at 500 nm was measured with increasing temperature at a rate of 0.18C / min. After cooling the sample to 208C in 20 min (the cooling rate was not precisely controlled), the heating was repeated for the next cycle. Thermal characterization of the polymers was carried out with a differential scanning calorimeter (DSC) (TA Instruments 2100) under nitrogen. The heating rate was 38C / min for dried polymer samples and 18C / min for swollen samples. Uniaxial tensile analysis was performed

2.6. Wound treatment

Water soluble PEO / PLLA multiblock copolymer (2000 / 1747, Mn 5 3208) (0.3 g) dissolved in 2 ml of distilled water was mixed with 0.2 (3:2 matrix) or 0.3 g (1:1 matrix) of water insoluble PEO / PLLA copolymer (2000 / 1678, Mn 5 48,864) dissolved in acetone. After applying vacuum condition for 20 min to remove most of the acetone, 1 mg of bFGF dissolved in 1 ml of PBS was added. After mixing the solution manually, it was cast and water was removed in vacuo for 2 h. The prepared lm was

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about 0.5 mm thick and was cut into 1 cm31 cm squares. Adult male SpragueDawley (SD) rats (200250 g) were acclimatized to our laboratory conditions for 35 days prior to use. SD rats were anesthetized with xylatine and ketamine mixed solution (50 mg / kg, intraperitoneal injection) and their dorsal hair was clipped. Four wound sites of approximate dimension of 1 cm (width)31 mm (length)33 mm (depth) were made on each rat by removing dorsal skin with a razor blade. A cast lm containing bFGF was placed onto the wound site of the rat skin surface after irradiation of UV light for 10 min for sterilization [17]. Wound sites without any treatment and covered with a commercial wound ointment (Fucidate) were considered as controls. The wound recovery rate was monitored visually.

succinic acid, and was inversely proportional to the amount of succinic acid added.

3. Results

3.1.3. Multiblock copolymers The formation of alternating multiblock copolymers was conrmed by IR and 1 H-NMR spectra (Fig. 1) and also by molecular weight measurements using GPC. The 1 H-NMR spectrum shown in Fig. 1 ascertains the chemical composition of PEG / PLLA multiblock copolymer. The peaks at 5.1 ppm (CH) and 1.5 ppm (CH 3 ) belong to PLLA blocks and the peak at 3.6 ppm is characteristic of main chain methylene units in the PEG blocks. The a-methylene protons of PLLA-connecting ethylene glycol units (PLLACOOCH 2 ) appear at 4.25 ppm, together 2 with CH protons from the hydroxylated lactyl end units. Methyl protons in these end units appear at 1.25 ppm and are distinguishable with other methyl groups. The peak at 1.8 ppm may be from impurities of DCC or water. 3.2. Thermal analysis
Although ordinary PLLA showed a sharp melting peak around 1608C, the PLLA synthesized in the presence of succinic acid showed broad and complex melting endotherms below 1208C. When PLLA was incorporated into a block copolymer (typically, PEO / PLLA 2000 / 1990, Mn 5 25,000), the PLLA melting was further decreased to around 708C as shown in Fig. 2. The low crystalline melting temperature (T m ) of dicarboxylated PLLA synthesized in this study may be caused by irregular chemical structure originating from the succinic acid residue in a relatively short PLLA and the resulting defects in PLLA crystallinity. The phase mixing of PLLA with PEO segments in the block copolymers further decreased PLLA T m to around 708C. The melting of PEO in the block copolymer appeared at 278C. PLLA crystallinity remained after swelling in water and PLLA melting appeared at the same temperature, while no PEO melting was observed due to PEO hydration. However, when the swollen gel was rescanned in DSC measurement, no clear PLLA melting peak appeared. A similar pattern of PCL melting of PEO / PCL multiblock copolymers was observed. In dried and swollen samples, PCL melting was found around 408C.

3.1. Synthesis and structure characterization

The chemical structures of dicarboxylated PEO, dicarboxylated PLLA, and PEO / PCL and PEO / PLLA block copolymers were identied by FT-IR and 1 H-NMR spectra.

3.1.1. Dicarboxylated PEO It was found that every PEO chain contains one molecule of DMAP. IR (KBr tablet): 1735 (C=O), 1645 (C=N 1 ), 1559 (COO 2), 1115 cm 21 (CH 2 CH 2 O). ] ] 1 H-NMR (CDCl 3 ): d 4.2 (OCH 2 CH 2 OCO); d 3.7 ] (CH 2 CH 2 O); d 2.7 (COCH 2 CH 2 COOH); d 3.2 ] ] ] ] ((CH 3 ) 2 N of DMAP); d 6.68.3 (pyridine-H of ] DMAP). 3.1.2. Dicarboxylated PLLA The characteristic peak from succinic acid residues, CH 2 CH 2 , appeared at 2.70 ppm. 1 H-NMR spectra of PLLAs were used to determine the molecular weight of the prepolymers from the integration ratio of the CH peak from lactic acid residues at 5.15 ppm to that from succinic acid residue. The molecular weight of dicarboxylated PLLA was varied by changing the feed ratio of L-lactic acid and

Y.H. Bae et al. / Journal of Controlled Release 64 (2000) 3 13

Fig. 1. A typical 1 H-spectrum of PEO / PLLA multiblock copolymer (PEO / PLLA 2000 / 1675, Mn 5 48,864).

3.3. Swelling studies

Fig. 3 shows the swelling changes of PEO / PCL multiblock copolymers between two xed temperatures. When the initially dried samples were placed in a pH 7.4 PBS solution at low temperatures, the equilibrium swelling was reached within 4 h. After 24 h, exposure to high temperatures which were around or above PCL melting temperatures, the degrees of swelling reached their new equilibrium levels. When the temperature was lowered to the initial lower temperatures, all samples showed increased swelling levels rather than decrease in swelling. After the rst temperature cycle, the swelling was reversibly changed between the two temperatures, low swelling at high temperature and high swelling at low temperature. The molecular weight (Mw ) of the constitutive PEO and PCL blocks determined the swelling levels of the polymers at the two temperatures. However, the swelling changes of PEO / PLLA block copolymers were not reversible

Fig. 2. DSC thermograms of PEO / PLLA 2000 / 1990 (Mn 5 25,000) multiblock copolymer.

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Fig. 3. Swelling changes of PEO / PCL multiblock copolymer in response to step-wise temperature uctuations between two xed temperatures. d, PEO / PCL 1500 / 2000 (Mn 5 38,000) (15 508C); j, PEO / PCL 1000 / 1250 (Mn 5 28,000) (15378C); m, PEO / PCL 1000 / 2000 (Mn 5 37,800) (20608C).

Fig. 5. The weight swelling ratios of PEO / PLLA multiblock copolymers with various block lengths and molecular weights of the polymers in pH 7 buffer solution at 378C.

even after repeated thermal cycles as shown in Fig. 4. Fig. 5 demonstrates the effects of polymer composition and molecular weight of PEO / PLLA multiblock copolymers on equilibrium swelling levels at 378C. With a xed PEO block size and with similar

Mw , swelling decreased as PLLA length increased. Swelling was also strongly inuenced by the Mw of the polymers.

3.4. Mechanical properties

Block copolymer lms prepared by the solvent casting method were rubbery in both dried and swollen states. PEO / PLLA lms showed a reasonable but not remarkable mechanical strength (Youngs modulus) of around 1 MPa in dried and swollen states. The Youngs moduli of PEO / PCL samples in various states; dried, swollen in water at 238C, and swollen at 238C but exposed above its PCL melting temperature, are summarized in Fig. 6. The Youngs moduli of the polymers were found to be 23 MPa, 45 MPa, and 30 MPa for PEO / PCL 1000 / 1250 (Mn 5 28,000), 1000 / 2000 (Mn 5 38,000), and 1500 / 2000 (Mn 5 38,000), respectively. Considering that the Youngs modulus of Pellethane (a typical biomedical polyurethane) is about 18 MPa [18], the mechanical strengths of the block copolymers were remarkable. The Youngs moduli of swollen samples were about 3040% of those from dried samples. When the test was performed after exposing the samples to above the melting temperature of PCL domains followed by cooling, the

Fig. 4. Swelling changes of PEO / PLLA 2000 / 1690 (Mn 5 48,900) multiblock copolymer in response to step-wise temperature uctuations between two xed temperatures.

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Fig. 6. Youngs moduli of PEO / PCL multiblock copolymers at different states. h, dried; , swollen at 238C; , swollen at 238C but after exposure to 608C for 20 min.

copolymers in controlling molecular weight, amphiphilicity, swelling, mechanical properties, and probably degradation. However, few investigations of biodegradable multiblock copolymers have been reported in the literature. In this study, to obtain alternating multiblock copolymers constituted of PEO as a hydrophilic block and PLLA or PCL as a biodegradable block, functionalization of PEO and PLLA at both chain ends with carboxylic groups was attempted, followed by a coupling reaction. The carboxylation of PEO and synthesis of dicarboxylated PLLA resulted in functionality .95%. In step growth polymerization, the governing parameters in determining the molecular weight of resulting polymers are functional group ratio (r) in the feed (e.g. [OH] / [COOH] in this experiment) and the extent of reaction ( p, conversion). The relationship of these parameters with number average degree of polymerization (DP) is expressed as: DP 5 (1 1 r) /(1 1 r 2 2p) When functional prepolymers having average Mw of 1000 were used, a number average degree of polymerization (DP) larger than 20 will be required to produce a polymer of Mn . 20,000, which is a reasonable criterion for mechanical properties. Since we were able to obtain functionality over 95% for dicarboxylated PEO and dicarboxylated PLLA, as long as the conversion goes above 97.5%, a polymer of DP520 can be obtained. With higher Mw prepolymers, it is more feasible to construct alternating multiblock copolymers of high Mw . The number average Mw (Mn ) of various multiblock copolymers synthesized in this experiment ranged from 12,300 to 50,000 with polydispersity of 1.31.5 and they formed thermoplastic hydrogels. The lowered melting temperature of the hydrophobic segments of PCL and PLLA, which was caused by the presence of di(ethylene glycol) residue and succinic acid residue in the prepolymers, respectively, and by phase mixing with PEO segments in the block copolymers, make it possible to process the polymers at low temperatures. This is important because high temperature processes for fabrication of desired products could initiate the thermal degradation of polymers and could also have deleterious effects on incorporated active ingredients.

strength decreased to less than 20% of those before exposure to high temperature.

3.5. Wound covering

The healing of wounds covered with bFGF incorporated PEO / PLLA block copolymer lm fabricated with the mixture of low and high Mw s of PEO / PLLA block copolymer (low Mw :high Mw 5 3:2 and 1:1, respectively) is presented in Fig. 7 with the control experiments of coverage with a commercial wound covering ointment (Fucidate), and without treatment. Wounds which had been covered with bFGF lm showed complete recovery after 10 days, while the control wound sites were only partially healed. This result proves that the PEO / PLLA lm released active bFGF at the wound site, while the only multiblock copolymer was less effective on the wound site of a rat model (data not shown).

4. Discussion The alternating multiblock copolymer approach using short difunctional prepolymers may offer more benecial aspects over corresponding di- or triblock

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bFGF & PEO/PLLA matrix (low mw : high mw = 1 : 1)

No treatment

bFGF & PEO/PLLA matrix low mw : high mw = 3 : 2) Dorsal skin wound sites on a rat (day 0)

Ointment (Fucidate)

After wound treatment (day 0)

After 10 days

Fig. 7. Healing of wound sites covered by bFGF incorporated PEO / PLLA multiblock copolymer lm, commercial ointment (control), and no treatment (control).

In multiblock copolymers, the biodegradable blocks are connected by PEO. The solubility of high Mw block copolymers corresponds well to the solubility of the low Mw hydrophobic polymers, considering that low Mw polymers are more readily soluble in solvents. Moreover, PEO, which is soluble in a wide range of organic and aqueous solvents, endows additional solubility due to the presence of PEO segments next to the biodegradable blocks. This feature makes the multiblock copolymers synthesized soluble in various volatile solvents and in water / ethanol and water / acetone mixtures with the volume fraction of the organic solvents .60% v / v. This is important whenever a non-toxic solvent is required for processing purposes such as solvent casting or microsphere production. Another interesting point is

that the polymer solution can be made in a sterile solvent such as 70% ethanol solution, obviating the need for additional sterilization procedures after product fabrication. The thermoresponsive swelling of non-degradable polymers such as poly(N-isopropylacrylamide) and related copolymers has been thoroughly investigated for pulsatile drug release [19]. However, biodegradable hydrogels have attracted little attention concerning temperature responsive swelling. PEO / PCL multiblock copolymer demonstrated unusual swelling behavior in response to step-wise temperature changes. When the polymer is exposed to water at low temperatures, PEO segments were hydrated while PCL crystallinity was maintained. After reaching equilibrium swelling, when the swollen gel was

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exposed to PCL melting temperature, the swelling level was elevated by thermally disordered PCL crystallinity. The interesting nding was that the swelling increased further, rather than decreasing, when the temperature was lowered to the initial lower temperature. This observation may be explained as follows: when the PCL segment loses its crystallinity with temperature change in the presence of water, it allows interactions with water molecules, leading to water penetration and increased swelling. Afterwards, PCL does not recrystallize even at low temperatures within the experimental time scale for temperature changes. Rather, the PCL segments are associated by hydrophobic interaction, forming hydrophobic physical junctions. This hydrophobic interaction is weakened by lowered temperature, thus increasing temperature and elevated temperatures reduce the swelling level by enhanced hydrophobic interaction as demonstrated in Fig. 3. PEO / PLLA multiblock copolymers also showed the swelling changes by temperature cycles, where the high temperature was below PLLA melting temperature, but the swelling change was not reproducible. This may have been caused by the gradual phase mixing of both segments after repeated temperature cycles. This gradual phase mixing by the repeated thermal cycles can be indirectly supported by the light transmittance change of the polymer lm as shown in Fig. 8. The details of cycle dependent changes in transmittance of the lms was described elsewhere [20] which was inuenced by polymer composition and can probably be affected by partial degradation of the polymer. As demonstrated by the thermal analysis of the block copolymers, PLLA segments in the block copolymer formed loose crystals due to chemical structure and phase mixing with the PEO segment. Thus the polymer showed a Youngs modulus of about 1 MPa in dried and swollen samples even with high Mw polymers, while the PEO / PCL multiblock copolymers demonstrated high tensile moduli, as seen in Fig. 6. This mechanical strength is signicantly reduced when the polymer is swollen, but its strength is sufcient and half of that of commercial Pellethane. However the strength decreased considerably after exposure to high temperatures above the PCL melting point. The high crystallinity of low Mw PCL segments may be responsible for the unusual mechanical properties. After melting, the

Fig. 8. Thermal cycle dependent light transmittance of PEO / PLLA 2000 / 1690 (Mn 5 30,000) lm as a function of temperature in pH 7 buffer solution.

weak hydrophobic association degrades the mechanical properties. The unique properties of multiblock copolymers could be tailored for specic applications by changing hydrophobic blocks and manipulating temperature. An attempt to use the PEO / PLLA multiblock copolymer in a wound healing application was made. bFGF was loaded in the PEO / PLLA matrix which was fabricated by solvent casting of a mixed polymer solution of low and high molecular weight polymers. The low Mw polymer was used for wound adhesion property and the high Mw was for lm integrity and strength for handling. When the bFGF-impregnated matrix was applied to the wound, the healing rate was faster than other control sites with no coverage and commercial ointment containing fucidic acid. Although the quantitative analysis is under investigation, the preliminary results showed that the bioactivity of bFGF is preserved at least partially in the matrix and released in an active form to effect the wound healing process.

5. Conclusion Biomaterial design using amphiphilic multiblock

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copolymers is an attractive approach to manipulate swelling, thermo-responsive swelling, mechanical property, solubility, and probably degradation rate. The nature of the hydrophobic block used, polymer composition, molecular weight, and thermal history inuenced these properties. When PEO / PLLA multiblock copolymers were applied as a wound healing material loaded with basic broblast growth factor (bFGF), the feasibility study showed improved wound recovery when compared to controls of no treatment and the same wound covering without bFGF, indicating that a certain degree of the bioactivity of bFGF is preserved.

[8]

[9]

[10]

[11]

[12]

Acknowledgements
[13]

This work was partially supported by a special fund for K-JIST from the Ministry of Science and Technology and by grant no. HMP-95-2-22 from the Ministry of Health and Welfare, Korea. The authors thank J.-E. Jung for her technical assistance.

[14]

[15]

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