4 Blomatr Sk. Polymer Bn No. 2. No. 127-124 2001) BNSPDOI Porous starch-based drug delivery systems processed by a microwave route P.B, MALAFAYA™, C. ELVIRA", A. GALLARDO?, J. SAN ROMAN? and R.L. REIS! "Deparment of Por Engineering, School of Engineering, Universi of Minho, ‘Campus de Guat, 4710057 Boge, Portugal 2 Inte of Pobmer Slence and Tehnology CSIC cf Juon dela Ciera 3 ‘28006 Madi. Spin Received 31 August 2000; accepted 21 Angust 2001 Abstract—A new simple processing rut o produ starch-tased porous materials was developed ‘red on amicrowave ting ehodclogy, This inmvative procesiag rut Was sod to obi 0 leaded onl nd loaded rg delivery cares, nerpocating soe teri ant-inflammatry agen. “Tis blonctve agent ns sees ssl Ug wal exyetatins that he developed methoelogy tight be wid footer regs end gromth fcr Te prepared stems were charcerized by "Hand BENMR spetosopy whch allow the sy of the teractions between the srt based terials and te processing compaents, the Bowing agents. The posi of the prepared mails was ‘stnted by messuring thei apparent dest ad sued by comperng dru loaded and nowlouded ‘aries. The Sehavior ofthe porous sutures, wile immersed in aqueous media was ted in terms of swelling and gration, being intimately related ot presi. Pinal i iro drug {ele sts were perfonned showing a clear br effect followed by a slow onrled release of the dg over vere ays up 010 ay), Key word Starch ares; rug delivery systems: Bolegradable; porous syste INTRODUCTION Starch is a natural polymer that presents excellent characteristics to be applied in the biomaterials field, which are mainly low toxicity, biodegradability, and good stability (1]. Some starch-based products are already commercialized and many others are still under investigation, mainly due to the control of the degradation process of starch when in contact with body fluids or tissues (1) “To whom crespondeoce shouldbe added, Ean: palafay@ dep aminora PB Mlsfaya ea ‘Several attempts to produce porous biodegradable polymeric biomaterials have been caried out. The developed systems have been applied as drug delivery carers ‘or tissue engineering scaffolds, being produced by methods suchas particle leaching in poly-HEMA [2] and poly-urethane systems (3), gas emission in alginate (4), ot thermal treatments of chitosan [5] Inthe field of oral drug delivery, starch based materials have been investigated mainly as excipients for the manufacture of solid oral dosage forms, such as linear short-chain starch products prepared by pre-gelatinization of potato starch ‘which is proposed as an excipient for pharmaceutical tablets for the delivery of, theophylline [6], and cross-linked high amylose starch developed as excipient for the formulation of contolled release of solid dosage forms, Contramid™ (71 Furthermore, a combination of microcrystalline wax and starch derivatives have beet! used to formulate sustained and immediate Tbuprofen™ release formuls- tions (8). Degradable erass-Linked starch microspheres (DSMs, Spherex™) have been also applied as embolic agents, which are now in clinical use in chemother: peutic processes [9]. Several olhers types of administration of starch-based de- livery systems are still under investigation, such as nasal and buccal delivery of biodegradable microparticles (10), intravenous administration of polymerized ‘potato slarch microspheres [11], and subcutaneous implantation of cross-linked amylose starch (12) Iinthis work, a new simple processing rte to produce a sarch-based porous drug delivery system was developed. One innovative feature that ean be very useful in the clinical area i thatthe drug is loaded during the processing, allowing fora more homogeneous dispersion ofthe drug within the polymeric carrer and a controlled release which depends on the porosity cf the prepared systems, as well as on their swelling and degradation behaviours. The developed systems are aimed at being used as parenteral or implantation systems. This arile presents in vitro data only: the in vivo fate of the polymeric carriers will be considered in future studies. MATERIALS AND METHODS Materials ‘The polymer used in this study is based on a blend of com starch/ethylene vinyl alcohol blend (SEVA-C). The blend coatains around 50% starch by weight, while the copolymer discloses a composition of 60/40 mol/mol. SEVA-C was supplied by Novamont, Italy. The material has already been shown to be biodegradable and biocompatible [13-15]. The proposed methodology can also be used for other blends of starch with synthetic polymes, ic. cellulose acetate, poly eaprolactone and poly lactic acid, which are being studied in our research group. Although in the present paper we have only reported results for blends of starch with EVOH, the selection of other synthetic polymers forthe blends might allow the development of, fully-degradable cariers.Porous starch-baed dr delivery ystems m9 Unmodified native corn starch containing approximately 73% amylopectin end 27% amylose, from Sigma Chemical Co. was alo used. The naive com starch was used to study the effets of the hydrogen peroxide (H;0;) during the processing route and to investigate the possible interactions between the starch and the rug ‘A commercially-avilable baking powder, containing com starch, sodium py- rophosphate and sodium bicarbonate, from Nabisco Ibéria Ltd, Spain, was used as blowing agent (BA). The drug usd in the present study is meclofenamic sodium salt (MS) (CidHioCiaNN«O;) fom Puke Davis, which is an ant-inflammatory agent, melting point 289-291°C, and solubility in water of 15 mgmt Processing route ‘The processing route for producing the degradable polymer carers to be used as deug delivery systems was based, as stated before, on the utilisation ofa baking Povider a an additional BA. A fixed amount of HO; (30% v/v) was also used 48 a BA for the production of the polymeric carirs. ‘The BAS were mixed with the SEVA-C powders in amounts of and 10 wt. H,O2 was then added until a consistent sluny dispersion was obtained. The adequate quantity of HO: was frst optimized, then fixed at 1:1 (wt/wt, with respect othe total weight of bend, BA, and dug in the samples) and subsequently the same amount was used for the prodoction of all the samples (see Table 1). ‘The hydrogen peroxide also sets a BA, and consequently, some sales without any baking powder (only ‘With 1,02) were also produced, being possible to study the effets ofthe HzO and is inerations withthe drug. the production ofthe drug delivery systems, the drug was added in amounts of 20 ww, dissolved in H,O:, during the processing route as described above. ‘The mixture was then processed in a microwave oven. The oven power setup and treatment time was optimized as a function of several conditions such asthe presence of aking powder and ofthe drug. Typical values ‘Table. Nomenclature and porosity of the bined materials SVR MOOS —OMIO MMO OMS MIO SAC 100 9S ~~ SCSSCSSSSCS Bat = = 5 5 pn = 2jj- £5 20» Ho! 10 re e 128 sz 036 029k aS Porosity (@) 0. 0s) 0m Om 06s es 8 Wel perentager ofthe sold warting marl SEVA.C + BA dug). 5 The same amount of mililies of HO: slutonas grams of solid part added «Ral density ia the ease of SEVA-C and apparent density forthe ested materials * nested material withoot dr nd BA,1230 PB. Melfoye were between 250 and 400 W for the furnace power and 1 and 5 min for the treatment time. The samples were then dried at $0°C for 1 b in another oven. ‘The samples were prepared in a cylindcal shape with a diameter of 34 mm and ‘an average height of 10:2 mm. The samples were then cu into equal sub-samples cof 25 x 10 x 5 mm rectangular shape tht were usd in all the experiments ‘Nuclear magnetic resonance (NMR) and Fourier transform infra-red spectroscopy (FIR) analysis Nuclear magnetic resonance (NMR) and Fourie transform infrared spectroscopy (FPR) analysis were carted out in onder to characterize the materials, study the eles of 10> during the processing out, and to study the possible interactions berwieen the starch and the drug “The obtained materials were analysed by means of Hand "C-NMR specoscopy ina Varian XLR-300 specuometer. The proton specta were recorded on 5% (W/¥) deuterated dimethylsulfoxide DMSO-d, solutions, wile the carbon spectra were recorded on 15% (w/¥) deuterated dimehylslfoxide DMSO-d solutions a 70°C with the spectrometer operating at 75 MHz, using a flip angle of 90 deg (pulse ‘with of 13 1), arelaxation decay of 4s, and with inverse-gated decoupling in the acquisition to ensure the complet intensities of all the \C nuclei analysed. FTIR spectra were recorded on a Nicolet $20 spectrometer. Spectra were taken with a resolution of 2 cm" and were averaged over 120 scans. Samples ‘were thoroughly ground with exhausively-, the porosity can be obtained, as an approximate estimation, from this expression: = 1- pap/pe Aqueous behaviour ‘The water uptake ofthe samples was exaluated by immersion on an isotonic saline solution at 37 1°C (NaCl 0.154 M4, pH = 7.4 + 0,02), by means of measuring the weight of the samples after several prefixed soaking periods. ‘The assaysPorous starc-bosed dg delivery ystems vst were carried out until the water uptake tends to stabilize or until total or partial disintegration of the samples occurs. Ths typically corresponded to periods ranging from 24 t0 240 h. ‘The degradation assays were carried out in vitro, again in an isotonic s solution (NaCl 0.154 M, pH = 7.4 + 0.02}. Each sample (25 x 10 x 5 mm) was Soaked in 20 ml of test solution. The containers were carefully sealed and placed in ‘thermostatic bath at 37 °C. Test periods were 1, 2,3, and 4 h, and 2, 4, 7, 1, and 21 days. After each test perio, the samples were removed and dried to constant ‘mass. The percentage of weight loss was determined by weighting the final mass of the samples. Release profile In vitro drug release tests were carried out in isotonic saline solutions (NaCl (0.154 M, pH = 74 4 0.02). The samples (25 x 10 x 5 mm) were placed in ‘containers carefully sealed with 20 ml of test solution at 37 + 1°C for periods up 10 10 days, Samples were fully immersed providing total contact between their entire surface and the test solution, At different periods, solutions aliquots of 5 ml were removed, and then the same initial volume was adjusted with new fresh solution ‘The drug release was measured over a period of 10 days by UV spectrophotometry ina Perkin-Elmer 554, taking the original test solution as reference. RESULTS AND DISCUSSION ‘Nuclear magnetic resonance (NMR) and Fourier transform infra-red spectroscopy (FT-IR) analysis "Nuclear magnetic resonance (NMR) and Fourier transform infra-red spectroscopy (FTIR) analysis were used to characterize the materials, to study the effects of the HO: during the processing route and to investigate the possible interactions between the starch and the drug "THNMR spectra of native starch cor, starch reacted with HO inthe microwave conditions, and also with the meclofenamic sodium salt drug, are shown in Fig. 1 “The assignment of the starch signals (Fig. 1A) in the thee spectra are: 3.1 ppm 0 of solvent DMSO (2.5 ppm), 3.2-3.6 9pm ~CH,~ and —CH— groups at 43, 5.6, and 5:7 ppm the three hydroxyl groups, and at 5.4 ppm the anomeric ~CH— of sarch, Figure 1B shows the spectra of starch reacted with 1,03 under microwave ‘conditions. It is possible o observe a thin and intense signal at about 10.2 ppm, ‘which is in the range of aldehyde protons. However, in Fig. 1C, which corresponds to starch + HO, + drug reacted under microwave condition, the signal at 10.2 ppm signal is not present. Figure 2 shows the “C NMR spectra ofboth starch (Fig. 2A) and starch reacted with HO, (Fig. 2B). Ic is possible to assign the signals to the carbon atoms ofFigure. !11NMR specif (A) starchy (B) starch HO; and (© stch + HO» + eclfenamic sun salt (MS). the starch unis. Figue 2A shows the spect of starch + HO teated under mjcrowave conditions where it canbe observed as an intense and thin signal i the cabony! region (160 ppm with respect to TMS), and signals at 70, 67, and 48 pr, ‘hich compared tothe broad reference golymer peaks and can be assigned to low molecular weight products of the starch oxidation reaction. FTIR spectroscopy reveals the appearance of shoulder inthe carbonyl region at about 1740 em! for the starch reacted with HO2 under microwave conditions. Tecan be deduced that the treatment of starch with HO under these conditions Teads othe partial oxidation ofthe starch hydroxy] groups to aldehyde groups withPerot tarch-based dr delery systems 33 ie a eee ee le hal fear ae ee ee ee igure 2 "CNM spec of (A) starch; and (B) starch + HeOs ‘ring opening, and o products of low molecular weight, s has been described for the oxidation of starch with periodate sats [16, 17]. ‘An interesting aspect of using the proposed procedure to prepare porous systems based on starch to be used as drug delivery carriers is thatthe incorporation of the meclofenamic sodium salt produces an anti-oxidant or inhibitor effect in the oxidation of starch by H30:, as no signals assigned to oxidation products can bbe observed in NMR spectra, This can be atributed to the stach—meclofenamic1 PB. Mlafaya ea teractions through the polar hydroxyl, carboxylic, and amino groups by hydrogen bonding which make these systems more stable o oxidation by HO. Structural characterization: porosity “The microwave blowing method leads to porous materials, as it can be seen in the micrographs of Figs 3 and 4. The porosity was estimated by measuring the apparent density, fy, of the samples, as described above. ‘The porosity values obtained by this technigue are quoted in Table 1 together ‘with the apparent densities. It has tobe stressed that these are approximate values, particularly in the case of the loaded materials which contain a mixture of SEVA-C. ‘and drug, with the real density slightly differen to that of pure SEVA-C. However, these results can be used for comparative analysis and have been represented in Fig. 5, with zero porosity being taken as reference that corresponds to the untreated material. Since the amount of H;O3 is much higher than the amount of BA (1:1 viwtand 5 or 10 wt% respectively, the blowing effect assigned to the HO is cleatly higher than the effect of the BA as deduced from the porosity data Moreover, as should be expected, the porosity increases in both series (loaded and non-foaded) withthe amount of BA, although ths effect is more pronounced in the ‘case of the non-loaded series (M). Figure 3. SEM micrographs of (3) MO (SEA) MS (GSEVAC + 1o%wr Bay, SEVA.C + Sut BA); and (e) MIO3s @ Figure 3. (Contiua ‘Aqueous behaviour: swelling and degradation ‘The aqueous behaviour of these materials should obviously be intimately related to their porous stricture. The higher the porosity of the structure, the greater the6 2B Malaya ta Figure 4. SEM micrograph of (a) MMO SEVA-C + 20%. MS): 6) MMS (SEVA.C + St BA‘ 20st MS): and (e) MMO (SEVA-C + Lt BA 420586" MS),Porous tarchbased drag delivery toms wa Figure 4 (Comins Figure , Porosity of the foded and nor tose poymeric systems surface area and water uptake capability. In addition, the higher should be the degradation rate and release rates (and the observed burst effec). Figure 6 shows the water uptake of the two series of materials. Non-porous SBVA-C flmshave an equilibrium hydration degree of 25% [18], whichis increased in all obtained porous systems, as can be observed in Fig. 6. In the case of the MO sample, the equilibrium hydration degree isthe lowest obtained as a result of the starch hydroxyl groups oxidation to aldehyde groups which are less hydrophilic, as has been previously discussed. The MM series (loaded) exhibit a clearly higher ‘water uptake than the M series (non-loaded). These observations can be attributed to the ionie character of the meclofenami sodium salt, which contributes to the1s PB. Mossy Water Uptake (%) Time (hours) igure 6, Water uptake ofthe onde and none ply ste. matrix being more hydrophilic. In this sens, it has tobe pointed out thatthe release studies (se Fig. 8) show a noticeable bust effect associated with a significant initial ‘weight loss (Fig. 7), and at 4 b (the time scale ofthe swelling experiments, the curulative release is 35-45% of the initial load (20 we). In addition 10 the intrinsic gravimetrical eror associated with this los, there is still an important residual ionic drug amount which makes the MM series strongly swellable with respect fo the M series during the first hours of immersion. Figure 6 also shows the aforementioned dependence on porosity, since the higher the porosity ofthe system (for each particular series), the greater the hydration degree ofthe materia ‘The degradation behaviour in aqueous media, expressed as weight loss vs time, is represented in Fig. 7 for both series. The samples exhibit a weight loss pattem similar to, bt kinetically greater than, that of SEVA-C non-porous materials {19}, ‘This is true forall samples except the MMO sample (the loaded sample processed without baking agent), which exhibits a lower degradation rate. This fact seems to show new evidence ofthe strong interactions between the drug and the starch for this particular sample and itis in agreement withthe NMR experiments where evidence ofthe preservative action ofthe drug on the starch oxidation could be found. These interactions seem to be weaker in the presence ofthe baking agent as the degradation rate is consistently higher. Thisis also related tothe effect ofthe drug on te blowing action of the BA. In fact, Fig. 5 shows that the blowing capacity of the BA in the M series is higher than in the MM series. It seems than the basic character of thePorous star-based dg delivery tens 1239 =-S =o Weight Lossi%) timo (hours) Figure 7, Degrdstion behaviour ofthe loaded an onload polymers systems carboxylate (which is the basic form of the carboxylic acid/carboxylate couple) fisplaces the equilibrium carbonate/bicarbonate/ carbonic acid through the basic species, and the consequently CO; formation is restricted. The erxor associated withthe drug release must be again stressed. This means for the MM series a total ‘weight loss for the fist days of 10%. From the data obtained on the degradation tests it is arguable to assume thatthe carriers willbe fully degraded in an adequate period of time. Of course the fate of ‘both polymeric phases should be studied ir: vivo. The available results published in the literature [14] indicate a clear in vivo degradation of the material, although itis not possible to have any indication of its complete resorption du to the relatively short implantation periods. However, we must again drav attention to the fact that the concepts and processing methodologies proposed herein might also be used for blends of starch with other undoubtedly degradable synthetic polymers. Also in some long-term release applications, for instance on carriers implanted in bony sites, it might not be disadvantageous to havea non-fully degradable system (if itis clearly biocompatible and does not disturb the surrounding tissues). But of course all these comments and speculations should be addressed in future studies Drug release ‘The in vitro cumulative release of the meclofenamic sodium satis depicted vs time in Fig. 8. There is clear burst effect with a fast release of half of the loaded drug (approximately) in less than 24h which is mainly associated with an initalm0 PB. Moifaya th 20 Eo 5 oo * === ahs ” Se intto ° $$ Time (nours) Figure 8, Releate profil bined on UV spectreicopy measurements othe sare basd polmeric rug ever systems fast weight loss, followed by a slow release over several days (up to 10 days) Inaddition, the macroporous nature ofthe material makes the accessibility of water tosome of the drug molecules easy, particularly to those that are close tothe surface. ‘The second step, characterized by a slow release, can be attributed to the diffusion of, the drug throughout the porous starch based structure. Finally, Fig. 8 shows again the dependence of the carrer porosity and that the release rate increases with the amount of BA. ‘CONCLUSIONS [New porous biocompatible and degradable porous starch-based blends were ob- tained by a simple and innovative mettodology. The newly-proposed processing route (based on a microwave processing) forthe production of the porous starch based drug delivery systems allows diug loading during the preparation of the porous delivery systems, This leads to a more homogeneous dispersion ofthe drug in the bulk of the entire carier. The data obtained from several characterization techniques indicates a partial oxidation ofthe hydroxyl groups of the starch macro- molecule by HO to aldehyde groups with ring opening and to products of low ‘molecular weight. The incorporation ofthe meclofenamic sodium salt produces an anti-oxidant or inhibitor effect in the oxidation of starch duc to HO. The different Porosity of the prepared systems, as well as drugstarch interactions are factors re-Porous sarh-based dr deliver systems waa sponsible forthe swelling and degradation behaviour of the porous starch systems, and consequently of the release profiles ofthe drug model which exhibit an initial burst effet followed by a slow continuous release over several days. [REFERENCES 1. L Hovgurd and H, Brondsted, Cri Ren Therp. Drug Carver yt. 18 185 (1990, 2. Gobi EL Hedberg, 2 Li, R, Babul, KK. Meslay an A. Nikos, Biomaterials 21, 2168 2000), 3, C.J, Spans, V- W Belgraer, 0. Rleasa, H,H. de Fro, RH. Veh and A.J Peaigs, Biomaterials 21,2453 2000 4, rete Yeh, RK Lata LD. She and. Moone, Biomaterials 2, 1921 (200), 5. S.¥, Madalyn H.W. Mabon, Biomaterials 20,113 (199), 6 G.HLP TeWierk, A.C. Eisen, J. Bergsma, AW. Areas Schl and C.F. Let, J Cont Rel 45,25 199). 7. ¥. Lene, L Moussa and . Duron, J. Cota. Rel. $3, 2251998). 1 F’Zhou,€ Vert, Me Scbelens, R, Lee and .P Remon, fet. Pharm 168, 79198), 9.1. Toga, Clin Pharmokinnis 26,75 (199. 10, SR Vyse and Inn J Microencap. 94571992) 1H. T Makita, H-Khimal. Kagab, K-Manba Naito, Lindblom, M. Habiouk nT, Oka, (Call Bik I. Rep. 13, 711983), 12 €:Déseraan, C. Girard, V- Lenears and P Dube, Proceed. Int. Symp. Control. Rl. Bio. Mater 26, 6391950). 13, $1 Medes, YP. Bowell, R. [Rebs CA, Bites al J.D. Belo in: 24h Anni Meeting Soin for Bizrate, San Diego, USA, 9 (1998). 14, $.€. Mendes, ¥ Bowel, RL. Reis, AvM. Cush apd J.D. Brij, Biomaterials 2, 2057 (oon, 15, MoE, Gomes, A.S. Ride, FB, Malaya, RL Res and A.M, Cubs, Biomaterials 2,883 (200, 16, AL. Poterand W-D. Hass J. Am. Chom So. 70,3774 (1948) 17, LA Wolf BT. Hoffer PR, Watsa, W.L Detherage and MM, MaeMser, J Am. Chem. Soc. 77 1654 (1955) 18, D. Demingor,C. Elvira J.F Mano, A M, Cath, EPiskin and RL. Reis, Pom. Degradation Sabi 7,161 2000). 19, COM, Var, RL Reis nd A.M. Cun, Ma es. movtions 2001 Gi pres)