Powder Technology 191 (2009) 155163

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Powder Technology
j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / p o w t e c

Fabrication of ultrane powder from eri silk through attritor and jet milling
Rangam Rajkhowa a, Lijing Wang a, Jagat Kanwar b, Xungai Wang a,
a b

Centre for Material and Fibre Innovation, Deakin University, Geelong, VIC 3217, Australia Institute of Biotechnology, Deakin University, Geelong, VIC 3217, Australia

a r t i c l e

i n f o

a b s t r a c t
Fibroin protein derived from silk bres has been extensively studied with exciting outcomes for a number of potential advanced biomaterial applications. However, one of the major challenges in applications lies in engineering broin into a desired form using a convenient production technology. In this paper, fabrication of ultrane powder from eri silk is reported. The silk cocoons were degummed and the extracted silk bres were then chopped into snippets prior to attritor and air jet milling. Effects of process control agents, material load and material to water ratio during attritor milling were studied. Compared to dry and drywet attritor milling, wet process emerged as the preferred option as it caused less colour change and facilitated easy handling. Ultrane silk powder with a volume based particle size d(0.5) of around 700 nm could be prepared following the sequence of chopping wet attritor milling spray drying air jet milling. Unlike most reported powder production methods, this method could fabricate silk particles in a short time without any pre-treatment on degummed bre. Moreover, the size range obtained is much smaller than that previously produced using standard milling devices. Reduction in bre tenacity either shortened the milling time even further or helped bypassing media milling to produce ne powder directly through jet milling. However, such reduction in bre strength did not help in increasing the ultimate particle neness. The study also revealed that particle density and particle morphology could be manipulated through appropriate changes in the degumming process. 2008 Elsevier B.V. All rights reserved.

Article history: Received 9 April 2008 Received in revised form 21 August 2008 Accepted 1 October 2008 Available online 17 October 2008 Keywords: Silk powder Milling Particle size Eri silk

1. Introduction Silk broin macromolecules have been extensively studied in recent years as biomaterial for a number of different non textile applications. This has been done through re-fabrication of native silk bres as nano bres [13], lms [2,4], hydrogels [5,6], foams [2,7], coating materials [8] and powders. Among these forms, silk powder is now produced commercially as an additive in cosmetic products and functional foods. Other potential applications of silk powder such as surface coating [911], bre treatment [12,13], llers in lms [14], ink [15] wound care [16], enzyme immobilisation [1719], composite scaffold for cell growth [20] and drug delivery [21,22] have also been reported. Powder can be prepared either by a solution route or by a mechanical method. In the solution route, silk bres are dissolved rst in a concentrated chaotropic salts followed by removal of the salts, and then powder from the aqueous silk solution is regenerated through a number of different means [17,2325]. The mechanical method, on the other hand, avoids lengthy, costly and environmentally sensitive production procedures associated with the solution route. It also retains the crystallinity and water insolubility of broin particles [26]. The present

Corresponding author. Tel.: +61 3 522 72894; fax: +61 3 5227 2539. E-mail address: xwang@deakin.edu.au (X. Wang). 0032-5910/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.powtec.2008.10.004

study adopted the mechanical attrition of bres to fabricate ultrane particles. Unlike inorganic materials such as metals and ceramics, organic polymer materials are very difcult to mill into ne particles. Milling of some synthetic polymers has been reported in the literature [2729]. Early works on silk milling used mostly ball and jet milling [14,26]. Long milling time (up to 2040 h) and restricted neness (d(0.5) = 10 20 m) remain the major issues in mechanical silk milling processes. To overcome these, various pre-treatments were used to reduce bre strength [26,3032]. Such an approach was also used for fabricating wool powder [33]. While the pre-treatments could increase milling efciency, they were not effective in reducing particle size to submicron scale. For some applications, sub-micron particles are likely to have advantages as they are highly reactive due to the high surface-tovolume (or mass) ratio [21,23,26,34]. Availability of such ultrane powder may also open out new application avenues of powdered silk. A method to prepare nano scale silk particles was reported [35], however, it relied on a special device using a high pressure of 1000 kgf/ cm2. The process also ltered large particles and used number based distribution to calculate the particle size, which is normally much smaller than the volume based calculation. Fabrication of sub-micron powder through standard milling devices is yet to be realised and we have made good progress in this direction in our recent study [36]. In the previous study, we discussed the silk fragmentation mechanism and milling kinetics using rotary and planatary ball milling [36].

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We obtained sub-micron particles through wet planatary ball milling using ne media, but subsequent drying caused particle agglomeration. In that study, the importance of friction in separating microbrillar architecture of silk was demonstrated through bre bending abrasion fatigue results. Use of process control agent (PCA) at the start of ball milling hindered the silk milling process as it reduced the frictional forces during grinding. We also observed that powder production remained inefcient without reducing bre tenacity to a certain extent via intensive degumming. Silk degumming is a chemical process that removes the glue like protein that surrounds the silk laments and holds two individual laments together. Preserving the native bre composition during powder production could be important for some applications. As an example, degradation during degumming sharply reduced performance of silk broin protein to the attachment and growth of cells in tissue culture experiments [37]. During the degumming process, some degradation and associated decrease in bre strength are unavoidable [38,39]. However, most of this strength can be retained following a standard degumming procedure used in this study. In the present study, an attritor mill was used to prepare ultrane powders from eri silk bre produced by Philosomia cynthia ricini silkworm. Unlike rotating container in a planetary ball mill used in our previous study, in the attritor, the balls are stirred inside a stationary container. This allows more irregular movement and spin to the media, resulting in a higher shear force and more frequent particle/ media collision than is the case for ball milling [40]. The use of attritor for ultrane and nano scale particle production and dispersion has been reported in recent years [4144]. Most of the materials processed on the attritor mill are, however, inorganic in nature and its suitability for soft organic materials such as silk bre has not been examined. In this study, air jet milling was also employed to assist further renement of the attritor milled particles/aggregates. Instead of using a grinding media, air jet mill uses the kinetic energy of the pressurised air, and comminution takes place due to impact between particles. Direct milling of silk snippets to powder using the jet mill only was also examined. 2. Experimental procedures 2.1. Material preparation Eri silk cocoons, which were open mouthed and free from pupae, were purchased from Northeastern India. They were degummed with a laboratory dyeing machine (from Thies) using laboratory grade sodium carbonate 2 g/L and sodium dodecyl sulphate (from Sigma Aldrich) 0.6 g/L at 100 C with a material (g) to liquor (L) ratio 1:25 for 120 min. The batch size of cocoons was 1 kg. To reduce bre strength for comparison trials, sodium carbonate concentration was increased to 10 g/L and temperature was raised up to 120 C in some degumming batches. 2.2. Powder production devices 2.2.1. Pre-milling A cutter mill (Pulverisette 19 from Fritsch) was used for chopping degummed silk. It has ve V shaped blades tted on a rotor, which rotates at a set speed of 2888 rpm. These blades in combination with three xed blades cut the bres until they became ne enough to pass through a 1 mm grid. In other words, this process reduced silk laments to approximately 1 mm snippets. Some snippets longer than 1 mm could also pass through the grid due to the small diameter of silk bres (1020 m). 2.2.2. Attritor milling Attritor (1S from Union Process) with a 9.5 L tank was used for both dry and wet grinding. Yttrium treated zirconium oxide grinding media

(5 mm) weighing 20 kg and with a bulk volume of 5.5 L was used for all experiments. Stirrer was set at 280 rpm. Cooling water (approximately 18 C) was circulated to minimise silk thermal degradation during attritor milling. 2.2.3. Air jet milling Sturtevant laboratory air jet mill with grinding air pressure 110 kg/cm2 was used. Material was fed using a powder hopper (K-Tron, made in USA) at a feed rate of 200 g/h. 2.2.4. Spray drying Dry powders from wet milled slurry were recovered through a laboratory spray dryer (B-290 from Buchi Labortechnik AG). The inlet temperature and slurry ow rate were 140 C and 78 mL/min, respectively. 2.3. Particle production Fig. 1 shows the schematic diagram of the milling sequence used in this study. Degummed silk was chopped into snippets for further size reduction. The snippets were ground either dry or with water in the attritor. In some wet milling experiments, snippets were pre-dry milled before wet milling. Powder from milled slurry was recovered using the spray dryer. Air jet milling was the nal step in powder production. Some snippets were also directly passed though the air jet mill without going through the media milling process. Preliminary dry and wet grinding trials were performed in order to determine the optimum loading of snippets in the attritor. Accordingly, unless specied otherwise, the snippet loading in all experiments was kept constant at 200 g standard mass per batch. During each experiment, if not specied otherwise, powder/slurry samples (b 50 mg) were collected at an interval of 1 h and particle size was measured. To reduce static charge during dry milling, 1% antistatic agent (ANLWS) based on weight of snippets was mixed with water (20:1 water to antistatic agent ratio) and sprayed over the snippets. Material was conditioned before milling at room temperature for at least 24 h to dry the sprayed water. Also, 15% (on weight of snippets) stearic acid (from Sigma Aldrich) or polyethylene glycol (PEG, MW 10,000, from Fluka) was added gradually either at the start or after a certain time (1, 2, 4 h) of milling as the process control agents (PCA) during dry milling. Stearic acid is commonly used as PCA to improve particle size reduction in metal powder processing. PEG is a biocompatible

Fig. 1. Schematic diagram of milling sequence.

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2.6. Measurement of colour change DIN6167 Yellowness and Ganz Grieoser Whiteness indices of powders were measured using a spectraphotometer SF600 Plus-CT (from Datacolor). Powders were compressed at a pressure of 2000 psi in a cylinder for 5 min to form 16 mm diameter discs. The discs were used for colour measurement. 2.7. Measurement of tapped density Powder samples were gradually lled in a Teon cylinder measuring 30 mm in diameter and 28 mm in height. The cylinder was tapped gently while lling the powder to allow the powder to settle uniformly. The cylinder top was levelled with a glass slide without applying any downward pressure. The weight of the powder was then measured and the density was calculated. Before measuring bre tenacity and powder tapped density, all samples were conditioned at 20 2 C and 65 2% relative humidity for 48 h. 3. Results and discussion 3.1. Dry attritor milling Fig. 2 shows particle size change during dry attritor milling and the inuence of PCA was added at different times of milling. It took only 2 h when the PCA was added at the early stage of milling or 4 h without PCA to fabricate powders with a volume d(0.5) of about 10 m. The milling time reduction is signicant compared to 1218 h required in an earlier study using planetary ball milling [36]. This difference may be attributed to the higher shear and impact energy provided by the attritor to brillate and fracture particles. It is known that a laboratory attritor works much more efciently than a conventional ball mill [44]. The result of the present study proved the same for silk milling. The result also reveals a different role of PCA in attritor milling compared to planatary ball milling. Previously, we observed that 3% stearic acid as PCA had a negative effect when added at the start of silk dry milling because of the reduction in friction which impaired brillation of snippets [36]. In attritor milling, results show improvement in milling performance with PCA whether it was added at the start or during milling, though less effective if added after 4 h dry milling. This result indicates that there is sufcient friction available in spite of the presence of PCA in the attritor milling for effective size reduction of silk. PCA may assist the process by preventing aggregation of small particles. However, it can be seen that PCA does not help much in increasing the limiting neness of particles. Effectiveness of PCA is reduced as particle size gets ner. There is also a possibility of PCA decomposition with time, which has been reported to occur during mechanical alloying of metal powder through ball milling [42,43]. The inuence of the quantity of PCA in dry milling was also studied. As seen from Table 1, adding stearic acid beyond 3% had only marginal benet. Considering possible contamination introduced by PCA, 13% addition of PCA appears to be a better choice. Polyethylene glycol which is biocompatible, may be effectively used for silk milling. It was observed that strong static electricity was generated during milling and as a result the milling media was covered by a thick layer

Fig. 2. Volume d(0.5) vs. time of milling of eri silk snippets (dry milling).

polymer and is also used as a PCA in inorganic material milling. It was therefore selected in view of the possibility of using silk powder for biological applications. Neither antistatic oil nor PCA was used when the material was drywet and wet milled. During wet milling, snippets/powder (g) to water (mL) ratio was 1:2.51:15. 2.4. Particle size measurement 2.4.1. Laser particle analyser Malvern Instruments Mastersizer 2000, which is a laser diffraction based particle size analyser, was used to measure particle size. Hydro 2000S side feeder was used to disperse particles for measurement. The dispersion medium was Propan-2-ol (from Sigma-Aldrich). The software (version 5.21) that came with Mastersizer 2000 was used for operation and data analyses. The software uses a least squares method with smoothness constraint (Tikhonov regularization) to solve the equations that connect size distribution with scattering energy. This helps regularisation and reduction of noise. The fundamental result of cumulative volume below a set of size intervals is tted by the software with a set of piece-wise cubic splines. Interpolation using the splines gives the frequency curve and median size. Refractive index of 1.542 and imaginary refractive index of 0.01 for eri silk were used for necessary calculations by the software. All particle size results reported are based on volume distribution of particles. As the powder samples produced are homogeneous, the variation between measurements within a batch is insignicant. Hence no error bars are plotted in d(0.5) measurements in this paper. 2.4.2. SEM Morphology of gold sputter coated particles was observed under a scanning electron microscope (LEO 1530 FEG-SEM) at 2 kV accelerated voltage and 24 mm working distance. SEM images of bre crosssections were also obtained to determine the bre linear density. 2.5. Measurement of bre tenacity Single bre tenacity was measured to determine the level of damage caused by degumming. A Single Fibre Analyser (SIFAN from BSC Electronics) was employed to measure bre breaking load using a jaw separation speed of 500 mm/min and a gauge length of 25 mm. The tested bres were sampled from the middle degummed cocoon layers. Their linear density, in dtex or gram per 10,000 m, was calculated from the measured mean cross-sectional area of cut laments collected from the same cocoon layers as used for tensile tests. Fibre tenacity was calculated from the bre breaking load and its linear density. A total of 100 silk bres were tested from each degummed batch.

Table 1 Effect of % of PCA on particle size reduction (PCA added at start) d(0.5) in m after 1 h 1% Stearic acid 3% Stearic acid 5% Stearic acid 3% Polyethylene glycol 21.4 14.5 13.3 14.9 d(0.5) in m after 2 h 11.5 11.6 11.0 10.3

158 Table 2 Colour results of attritor milled eri silk powder Material type Dry milled 6 h Wet milled 6 h Whiteness index (Ganz Grieoser) 55.8 71.9

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Yellowness index 22.4 12.8

of powder. It caused problems in separating ground powder from the media and collecting it from the milling container. The static electricity was due to the synthetic lining inside the cylinder wall and zirconium oxide sleeved agitating arm of the attritor, which provides an electricity insulating environment. In addition, abrading poor electrically conducting silk material during milling compounded the problem. Therefore, 1% antistatic oil was sprayed on the snippets to reduce the static problem. It was observed that PCA also helped to strip off powder from the media, but only up to a limit while it remains effective. Apart from the static build-up during milling, longer milling time increased the propensity of particles sticking to the media as they became ner. Strong adhesion to media inuences the movement of particles resulting in a reduction of the milling efciency and powder yield. In addition to static generation, poor thermal conductivity of bre material is also detrimental to dissipation of local heat generated during dry milling. Circulated cooling water through the attritor jacket though reduced the overall temperature inside the vessel to some extent, local heating probably caused some thermal degradation of powder material. It is clear from Table 2 that compared to wet milling, dry milling produces dull and yellowish powder. 3.2. Wet attritor milling 3.2.1. Wet milling of snippets Experiments were carried out to study milling kinetics and the optimum milling conditions in a wet milling process using silk snippets. 3.2.1.1. Snippets to water ratio. To optimise snippets to water ratio, tests were performed in the range 1:2.5 to 1:15. This range was selected based on some preliminary studies. A low ratio produces a thick slurry, which is difcult to drain out from the attritor and to subsequently spray dry. On the other hand, excessive water increases the volume, resulting in spilling out liquor from the system. The snippets to water ratio was changed by changing the quantity of water while keeping the snippets load constant at 200 g and the amount of media constant at 20 kg. Fig. 3 reveals that as the snippets to water ratio increases, the milling efciency improves rst, but then drops. Results in this study agree with the general trend reported with respect to solid content in the slurry during milling of some inorganic materials [4143]. It is reported that higher solid content leads to increased slurry viscosity which reduces kinetic energy of grinding media and as a result a lower milling rate is achieved. On the other hand, when it is diluted, though viscosity is reduced, collision frequency of particles drops as less particles are captured in the active grinding zone between the grinding beads. A good

Fig. 4. Fibre tenacity vs. wet milling time to fabricate particles with volume d(0.5) of around 5 m.

balance of collision intensity and collision frequency is required for good milling performance. For eri silk milling, the best results could be obtained using around 1:7.5 snippets to water ratio under the conditions examined. 3.2.1.2. Degumming condition. The tenacity of silk bre degummed through the applied normal degumming condition was 3.6 cN/dtex. The tenacity was drastically reduced with an increase in chemical concentration and/or temperature. The change in bre tensile strength has a signicant impact on the milling performance, as demonstrated in Fig. 4. Milling time to reach particle volume d(0.5) around 5 m drops at a linear rate with the drop in the bre tenacity as indicated from the three test results of milling conducted with bres of different tenacities. The decrease in milling time however does not ensure an increase in the limiting neness of particles. In fact, particle size still stabilized at around 5 m and remains independent to bre tenacity during wet milling. This probably happened due to particle aggregation and/or resistence to further fracture for the 5 mm grinding media used in these experiments. 3.2.1.3. Material loading. Silk snippets form a pulp when mixed with water. Owing to the thorough mixing and simultaneous reduction in particle size during the course of milling, large bre particles are ground into small ones, and the mixture becomes a homogeneous slurry. The time required to reach the slurry state varies depending upon the silk degumming conditions and material loading. A high material loading in the attritor will slow the media movement and the intensity of milling. The material mixure and the media movement can be observed through the feeding port of the attritor during the

Fig. 3. Effect of snippets to water ratio on particle size reduction in wet milling.

Fig. 5. Effect of snippets loading and degumming condition on wet milling efciency.

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Fig. 8. Change of volume d(0.5) after spray drying. Fig. 6. Volume d(0.5) vs. time of wet and drywet milling of normally degummed eri silk.

grinding process. Material state can also be observed by sampling and evaluating the product through the output port from time to time. As shown in Fig. 5, for normally degummed silk and 400 g snippets loading, the drop in d(0.5) in the rst 6 h was rather slow. The mixure still looked like a pulp, whereas after 12 h milling, the sample turned into a slurry. However, when reducing both snippets and water amounts by half without changing the grinding media, the material turned into a slurry during the rst hour. The dramatic change in milling efciency is reected in Fig. 5. Particles with d(0.5) around 10 m could be fabricated in 23 h. Hence, 200 g snippet load was used to carry out the rest of experiments using normally degummed silk. On the other hand, no such difference was observed while changing the snippets loading from 400 to 200 g when the bre strength was reduced from 3.6 cN/dtex to 0.8 cN/dtex by changing the degumming conditions. As seen in Fig. 5, the plots for 200 and 400 g loadings almost overlap for such intensively degummed low tenacity bre snippets. Though the snippet size was around 42 m compared to 63 m of normally degummed bres, the rate of size reduction for intensively degummed snippets was signicantly different and even 400 g loading could be milled rather easily. 3.2.2. Drywet milling As discussed earlier, dry milling caused a strong particle adhesion to the milling media which made collection of powder difcult after milling. Adhesion perhaps also contributes towards aggregation of particles effecting particle size reduction. A combined drywet process may alleviate this problem. In silk planatary ball milling, the drywet route achieved good milling results on low tenacity bres [36]. This combined process was therefore employed in this attritor milling to

improve particle neness prepared from regular tenacity silk. As shown in Fig. 6, the combined process reduced particle size further compared to wet milling only. Bimodal particle size distribution in Fig. 7 clearly demonstrates the presence of sizeable amount of submicron scale particles when the drywet combined milling progressed to 7 h. Continued milling up to 11 h, however, resulted in the aggregation of the ne particles represented by the corresponding single mode distribution plot in Fig. 7. An associated increase in volume d(0.5) at that time is also reected in Fig. 6. To overcome aggregation, slurry was diluted by increasing slurry volume by 50% with the addition of water and further milled for an hour. The return of the bimodal distribution is possibly due to particle de-aggregation. Though the process has an advantage over wet milling in improving particle neness, the dry milling phase caused signicant colour change. 3.3. Spray dried attritor milled particles 3.3.1. Normal tenacity silk Fig. 8 presents the change in d(0.5) during spray drying process. It can be seen that particle size does not change much upon drying when d(0.5) is approximately 5 m or larger. However, a difference is noticed on spray drying ner particles. Sub-micron scale particles if present in the slurry disappear after spray drying. An example from slurry prepared with single step wet milling of snippets is presented in Fig. 9. This indicates aggregation of ne particles in the dry powder. SEM images presented in Fig. 10 shed some light on the aggregation process. Silk bres are made of microbrils and the mechanical forces during milling can overcome the cohesive forces between microbrils and separate them [36]. The fragments of separated brils tend to aggregate into spheres. The 3 h milled SEM image in Fig. 10 shows that aggregates are made up of brous particles. As wet milling continues, dynamics of fragmentation, aggregation and de-aggregation yields a

Fig. 7. Particle size distribution of drywet milled slurry.

Fig. 9. Particle size distribution of wet attritor milled silk particles.

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Fig. 10. SEM images of spray dried eri silk particles after wet milling.

steady state with no further change in particle size. During this steady state, the irregular aggregates slowly become more spherical and compact as shown in the SEM image (Fig. 10) of 12 h milled particles. The size of a large number of primary particles in the aggregates is less than 1 m. Though these primary particles remain in aggregated form after spary drying, some of them are actually in a dispersed state in the slurry as demonstrated in the bimodal distribution in Fig. 9. These particles as they aggregate neither appear in the paticle size distribution after spray drying in Fig. 9 nor can be observed in the SEM image of dried powder in Fig. 10.

Fig. 12. SEM images of eri silk particles (intensively degummed).

Fig. 11. Eri silk snippet (intensively degummed).

While it is clear from Figs. 9 and 10 that a large number of submicron particles were produced in the milling process, it is not clear whether those in the range 110 m shown in the bimodal distribution (Fig. 9) are made up of only aggregates of ne particles or represent large primary particles with globular surface texture. Silkworm silk as well as spider silk form ne globular microsctructure with 100 200 nm globes at the broken tips during tensile fracture and the globes are conrmed as microbrils of silk [45]. A similar observation was also reported during pulling of silk bres under liquid nitrogen [46]. Such globular microstructure of fractured silk is attributed to the reversion of extended molecular chains when a force is applied to execute a fracture. During milling, bre particles are subjected to forces of

R. Rajkhowa et al. / Powder Technology 191 (2009) 155163 Table 3 Tapped density of spray dried particles Wet milling time 15 min 3h 3h Tenacity of degummed bres used (cN/dtex) 0.8 0.8 3.6 Tapped density (g/cm3) of spray dried particles 0.50 0.55 0.28

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different nature, including shear forces. It is possible that during the fracturing process, the microbrils roll up and form a globular textured silk particle from a single silk bre. The globular surface texture is shown in the magnied portion of the silk snippet in Fig.11. That part of the bre was subjected to some abrasion during the chopping process. 3.3.2. Low tenacity silk Size reduction rate on intensively degummed snippets during attritor milling was very fast as reected in Fig. 5. Particles with d(0.5) less than 10 m could be fabricated within 15 min milling (Figs. 5 and 12). Considering the diameter of eri silk which is around 15 m, it is clear that after 15 min milling, particles and fragments of splitted bres with globular microstructues are formed. As milling continued up to 30 min and beyond, the remaining brous particles were fragmented and then no obvious comminution took place. It is known that as particles become smaller further comminution is more difcult. However, the texture of particles becomes smoothened with continued milling. Such a change is probably due to the dominance of plastic behaviour of the intensively degummed silk material. The 3 h milled particles apparentlly look solid due to such surface smoothening process. The increased compactness is reected in the results of tapped density measurements shown in Table 3. The tapped density of normal degummed bre particles (with a higher tenacity) is much lower that that from the intensively degummed silk (with a lower tenacity), in spite of the increased milling time. The marked difference in particle size, shape and texture between silk powders prepared from normally degummed and intensively degummed bres is therefore related to the changes in viscoelastic behaviour of particles caused by a change in the degumming intensity. 3.4. Air jet milling 3.4.1. Air jet milling on snippets Table 4 shows that air jet milling on snippets is effective, if the bre strength is reduced by changing the degumming conditions. The molecular weight of silk is sharply reduced during a harsh degumming treatment [47]; accordingly a marked decrease in tensile and abrasion resistence can be observed after such a treatment [36]. Silk bre morphology consists of microbrils assembled in the form of pleated sheets [45,48]. Due to such morphology, separation of these sheets starts on the application of mechanical forces [49]. It happens much faster after severe reduction in bre strength [36]. Even during

Fig. 13. SEM image of eri silk (intensively degummed) directly air jet milled from snippets.

the chopping process, some fragments were formed (Fig. 11). The collision energy provided by jet milling could separate microbrillar sheets and produce ne powder once the bre strength was sharply reduced. As there is no media in air jet milling to help congregate these fragmented sheets into other shapes, ultimate particles remained aky in nature when jet milling was used directly on silk snippets. This is seen in the SEM image in Fig. 13. Absence of globular microstructure on the surface of these particles in contrast to media milled particles may be due to the lack of friction in air jet milling process. The direct air jet milling could be the preferred way of producing ne silk powder if changes in material resulting from harsh degumming conditions are acceptable. Apart from making the process shorter, avoiding the use of attritor eliminates any possible risk of contamination from the milling media as it is known that contamination may not be totally avoidable in media milling [44]. 3.4.2. Air jet milling of spray dried particles Aggregation in slurry during the milling process has been already discussed. One way of overcoming this and reducing silk powder size is the use of ner media. Using ne media for preparing nano dispersion of inorganic materials has been reported [43]. Our previous experiment also revealed better performance of ner media on silk milling [36]. However, ner media also increase the risk of contamination. Moreover, although particles are reduced to submicron scale during wet milling, it is difcult to obtain such submicron particles in dry form. Air jet milling was therefore used to

Table 4 Change in particle size on air jet milling Wet attritor milling time 0 (snippets) 15 min 30 min 1h 3h 0 (snippets) 3h 6h 12 h Type of degumming Intensive Intensive Intensive Intensive Intensive Normal Normal Normal Normal Volume d(0.5) in m Before air jet milling 42.5 9.8 7.4 6.6 6.8 63 8.9 5.6 5.6 After air jet milling 4.3 5.3 5.6 6.1 6.3 61 1.7 0.7 0.8

Fig. 14. Particle size distribution of air jet milled silk particles.

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explore the possibility of de-aggregation and fragmentation of spray dried particles to generate ultrane particles in dry form. It can be seen from Table 4 as well as in the distribution plot of jet milled particles in Fig. 14 that air jet milling can reduce particles of normally degummed bres signicantly and d(0.5) less than 1 m can be achieved. SEM image of such ne particles is provided in Fig. 15. Comparing this with SEM image before air jet milling in Fig. 10, it appears that air jet milling largely separates primary particles from the aggregates. However, jet milling could not provide a unimodal particle size distribution below 1 m as reected in the plot in Fig. 14. Some particles more than 1 m are visible also in Fig. 15. So the question remains as to whether or not the aggregates shown in Fig. 10 contained some large size primary particles. The result of air jet milling of intensively degummed bre pre milled by attritor is however diferent from normally degummed bre particles. It is seen from Table 4 that air jet milling failed to have noticeable impact on particle size reduction of intensively degummed and attritor milled silk particles. This is further conrmed by comparing SEM images between Figs. 12 and 16. Air jet milling worked well on brous shaped particles, but was not effective on attritor milled ne powder. The attritor milled particles were very compact and so could not be easily fractured. These results clearly indicate that a substantial reduction in bre tenacity provides a scope to cut down milling time signicantly. It also brings changes to particle shape and powder bulk density. At the same time, too low bre tenacity stands in the way of fabricating ultrane sub-micron scale particles. 4. Conclusions Fine silk powder can be fabricated in sizeable quantity using suitable standard grinding devices from degummed bres without any pre-treatment. Dry attritor milling was not a preferred approach, due to high static charges resulting in powders sticking to milling media, which reduce powder yield. From that consideration, wet milling following the dry milling was a better option. The combined process produced ner particles compared to single dry or wet attritor process alone. To avoid possible colour change during dry milling, wet milling only was eventually considered as the best option. By using 5 mm grinding media and appropriate amounts of material and material to water ratio, silk particles having a volume based particle size d(0.5) in the range 510 m can be fabricated within 36 h with wet attritor milling. During wet milling, brous shaped silk snippets turned into irregular sub-micron particles with time, but spherical aggregates of these primary particles were distinct, particularly after spray drying. Under the conditions examined, such aggregation was

Fig. 16. SEM image of intensively degummed eri silk particles: (a) 15 min attritor milled then air jet milled and (b) 3 h attritor milled then air jet milled.

responsible for stabilising the d(0.5) around 5 m in spite of longer milling time. Subsequent use of air jet milling to free the ultrane primary particles from the aggregates proved useful. The ultimate paricles with d(0.5) of approximately 700 nm could be produced from bres without harsh degumming or any other pre-treatments. Fibre tenacity played an important role in milling efciency and particle morphology. Low tenacity snippets resulted in a short milling time and a linear relationship between milling efciency and bre tenacity was recorded. With a substantial reduction in bre tenacity, highly compact particles were formed, and air jet milling was not effective to break these further into sub-micron primary particles. The study also revealed that, unlike normally degummed silk bres where media milling was necessary, low strength bre snippets could be directly converted by air jet milling into aky particles with a volume based particle size d(0.5) of around 5 m. References
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