Solvent casting-particulate leaching synthesis of a nano-SiO2/chitosan composite scaffold for potential use in bone tissue engineering

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This work aimed to introduce nanosized SiO2 into the CS matrix to produce three-dimensional (3D) scaffolds by solvent casting combined with salt leaching using NaCl as a porogen agent. The amount of the porogen to polymer was varied to produce 3D CS/SiO2 scaffolds with suitable pore sizes and porosity.

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Nội dung Text: Solvent casting-particulate leaching synthesis of a nano-SiO2/chitosan composite scaffold for potential use in bone tissue engineering

Cite this paper: Vietnam J. Chem., 2023, 61(5), 605-611 Research article DOI: 10.1002/vjch.202300020 Solvent casting-particulate leaching synthesis of a nano-SiO2/chitosan composite scaffold for potential use in bone tissue engineering Nguyen Kim Nga*, Tran Thi Thu Huyen, Ta Ngoc Dung School of Chemical Engineering, Hanoi University of Science and Technology, Hanoi 10000, Viet Nam Submitted January 27, 2023; Revised April 8, 2023; Accepted June 5, 2023 Abstract In this work, a three-dimensional (3D) scaffold comprised of chitosan and SiO2 nanoparticles (NPs) (CS/SiO2) was synthesized for bone tissue engineering. The scaffold was synthesized using a combination of solvent casting and salt leaching methods. The nanoparticle sizes were controlled by hydrothermal treatment with the aid of cetyltrimethylammonium bromide (CTAB), which was then used as a bio-inorganic component of the composite scaffold. Various methods, such as SEM, FTIR, XRD, and liquid substitution, were conducted to determine the morphology, structure, pore sizes, and porosity of the synthesized scaffolds and the interaction between the SiO2 and CS phases. The mechanical properties of the composite scaffolds were evaluated by testing their tensile strength. The results showed that the synthesized 3D CS/SiO2 scaffolds exhibited porous structures with suitable average pore sizes ranging from 198 to 269 µm and porosities from 70.99 to 73.23%, respectively. The tensile strengths of the CS/SiO2 scaffolds were around 1.57-1.83 MPa, matching well with those of cancellous bone. These appropriate values in terms of pore size, porosity, and tensile strength suggest that CS/SiO2 scaffolds could support cell migration, growth, and distribution. The synthesized CS/SiO2 scaffolds would be potential biomaterials for bone tissue engineering applications. Keywords. SiO2, scaffold, solvent casting, particulate leaching, bone-tissue engineering. 1. INTRODUCTION BTE scaffolds due to its high biocompatibility, non- antigenicity, and antibacterial properties.[8] Fabrication of bone tissue engineering (BTE) However, the low mechanical properties of chitosan scaffolds utilized in bone replacement and bone lead to the limitation of its use in BTE applications. healing has expanded rapidly over the last decades. The mechanical properties of chitosan can be In general, scaffolding materials have been designed improved by combining it with bioactive inorganics. to mimic natural bone structures for repairing or Previous studies have shown that silicon dioxide replacing injured bone.[1-3] A scaffold is a substrate (SiO2) or silica can support the crystallization of for implanted cells and the physical support in apatite crystals, cell adhesion, and collagen forming new tissue. Thus, an ideal scaffold needs to formation on scaffold surfaces.[9] Moreover, have biocompatibility and porous structure so that nanosized SiO2 has a large specific surface area and the cell can adhere and grow, tissue can develop, and can form a tighter interface with the polymer matrix tissue fluids and nutrients can transfer freely. in composites.[10] Thus, the combination of chitosan Usually, most scaffolds are composed of polymers, and nano SiO2 in composites is expected to enhance bio-ceramics, and hybrid materials.[4-6] Although tailored physical, biological, and mechanical numerous materials have been studied for bone properties for bone scaffolds. Several previous scaffold fabrication, only a few materials (e.g., nano- works have reported the successful fabrication of hydroxyapatite and biopolymer) have been reported CS-based scaffolds containing nano-SiO2 by the to be able to support cell growth within scaffolds.[7] freeze-drying method.[8,11] Kavya et al. have Thus, numerous studies focus on the fabrication of successfully synthesized chitosan/gelatin/nano-SiO2 composite scaffolds with a suitable microstructure to composite scaffolds by freeze-drying technique.[11] enhance cell adhesion and proliferation. Soumitri et al. also used this method to develop a Chitosan (CS) is an excellent biopolymer with bio-composite scaffold combining chitosan (CS), similar structural groups to the natural extracellular nano-scaled silicon dioxide (Si), and zirconia (Zr).[8] matrix. It can derive from the partial deacetylation of Their results showed that the presence of nano-silica chitin. CS is considered a potential candidate for improved the bioactivity and cellular compatibility 605 Wiley Online Library © 2023 Vietnam Academy of Science and Technology, Hanoi & Wiley-VCH GmbH
25728288, 2023, 5, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/vjch.202300020 by Readcube (Labtiva Inc.), Wiley Online Library on [01/05/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Vietnam Journal of Chemistry Nguyen Kim Nga et al. of the scaffolds better than those of the CS scaffolds method using TEOS as a precursor and CTAB was alone. Apart from freeze- drying method, several used as a template to produce the rod-shaped SiO2 methods have been used to fabricate 3D scaffolds NPs. The preparation and characterization of SiO2 including fiber bonding, phase separation, melt NPs were conducted in the same manner as in our molding, and solvent casting combined with previous work.[12] In this work, a specific amount of particulate leaching methods.[6,7] Among those CTAB (1.3 or 1.45 g) was used to further control the methods, solvent casting combined with particulate sizes of SiO2 NPs down to the nanosized similar to leaching showed high efficiency because the pore those of the mineral phase of bone. architecture of 3D scaffolds (e.g., pore size and 3D CS/SiO2 NPs scaffolds were prepared by porosity) can be easily controlled by varying solvent casting combined with a salt leaching porogen particle size and weight ratio of porogen to method with NaCl as the porogen. In a typical polymer without using the specialized equipment. experiment, 0.48 g of chitosan (CS) was dissolved in This work aimed to introduce nanosized SiO2 24 mL of 3% acetic acid for 3 h to produce a 2% into the CS matrix to produce three-dimensional (w/v) polymer solution. A suspension with 0.096 g (3D) scaffolds by solvent casting combined with salt of SiO2 NPs was prepared by ultrasonic vibration of leaching using NaCl as a porogen agent. The amount SiO2 NPs in 1.5 mL deionized water for 1 h. This of the porogen to polymer was varied to produce 3D suspension was then dripped into the above polymer CS/SiO2 scaffolds with suitable pore sizes and solution. The mixture was stirred on a magnetic porosity. It is known that the size and shape of a stirrer at a speed of 450 rpm for 1 h to achieve scaffold’s inorganic component play an important homogeneity. The homogeneous mixture was cast role in promoting cellular activities and into a 55 mm glass Petri dish containing various osteoconductivity of the scaffolds. The rod-shaped amounts of NaCl (16 or 17 g) with particle sizes of SiO2 particles were successfully prepared in our past 160- 250 µm. Afterward, the samples were air-dried work,[12] however, their sizes (average length of 231 under a chemical hood for 2 h and then in an oven at nm and average diameter of 113 nm) were 60oC for 36h. The resulting samples were immersed significantly larger than those of bone minerals. As a in 10% NaOH solution for 2 h and subsequently result, in this work, further studies were conducted washed several times with distilled water to leach to produce SiO2 NPs with similar sizes to bone out the salt. The scaffolds were further air-dried and minerals. The SiO2 NPs have been since used as an then stored for further analysis. For comparison, 3D inorganic phase to synthesize 3D CS/SiO2, applying scaffolds were also prepared in the same procedure, for bone tissue engineering. The morphology, without adding SiO2 NPs. Finally, four scaffold structure, and porosity of as-synthesized CS/SiO2 samples were obtained and labeled as CS1 and CS2 scaffolds were identified by FE-SEM, XRD, FTIR, for CS scaffolds with the salt amount of 16 and 17 g, and liquid substitution methods. respectively, and CS/Si1 and CS/Si2 for CS/SiO2 composite scaffolds with the salt amount of 16 and 2. MATERIALS AND METHODS 17 g, respectively. 2.1. Materials 2.3. Characterizations of SiO2 NPs and 3D CS/SiO2 composite scaffolds All reagents were of analytical grade and used as received without further purification. Acid acetic The morphological characteristics of CS/SiO2 CH3COOH, sodium hydroxide (NaOH), sodium scaffolds and SiO2 NPs were examined by a chloride (NaCl), ethanol (C2H5OH), 25% ammonia scanning electron microscope (FE-SEM, Hitachi- solution (NH3), tetraethyl orthosilicate (TEOS) S4800, Japan). The average pore diameters of the purchased from Merck; cetyltrimethylammonium scaffolds and average dimensions of SiO2 NPs bromide CH3(CH2)15N(Br)(CH3)3 (CTAB) (Sigma- (length and diameter) were measured from the FE- Aldrich); CS fakes exhibiting a degree of SEM images using ImageJ software. deacetylation of 85% and a low molecular weight The phase structures of the scaffolds and SiO2 were purchased from Nha Trang Aquatic Institute NPs were analyzed on a diffractometer (Panalytical (Vietnam). X’Pert-Pro) and their XRD patterns were recorded in a 2 range of 20°-70° with a scan step of 0.03°.s-1 2.2. Preparation of Nano-SiO2 particles, 3D under Cu-Kα radiation (λ = 1.5418 Å). Fourier CS/SiO2 composite scaffolds transform infrared spectra (FTIR) of the samples were conducted on a Nicolet 6700 spectrometer SiO2 NPs were prepared using the hydrothermal using the KBr pellet technique in the range of 4000- © 2023 Vietnam Academy of Science and Technology, Hanoi & Wiley-VCH GmbH www.vjc.wiley-vch.de 606
25728288, 2023, 5, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/vjch.202300020 by Readcube (Labtiva Inc.), Wiley Online Library on [01/05/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Vietnam Journal of Chemistry Solvent casting-particulate leaching synthesis of… 400 cm-1 with a resolution of 4 cm-1. The porosity of CTAB of 0.85 and 1 g (the lengths of 290±57 nm the scaffold was measured by the liquid substitution and 231±48 nm and widths of 142±21 nm and method using water as a liquid substitute and the 113±21 nm, respectively). Meanwhile, the SiO2 NPs formula used was as follows[13]: synthesized with the addition of 1.45 g CTAB (W2 − W1 )/ρw (Figure 1b) have irregular spherical and rod-like Porosity = shapes. It can be observed that these nanoparticles V2 − V3 where W1 is the initial weight of the dry specimen show such a small and rod-like shape that they with a 10 mm × 10 mm dimension, before agglomerate together to form clusters. Thus, it is immersing in water in a graduated cylinder. W2 is its difficult to determine the mean sizes of these SiO2 final weight after immersing in water. V2 is the NPs. The results confirmed that the amount of volume recorded after immersing the specimen in CTAB added to the synthesized mixture has a water. V3 is the volume of the residual water after significant effect on the sizes of the SiO2 NPs removing the water-impregnated specimen from the formed. 1.3 g of CTAB should be preferable to graduated cylinder. produce the smallest rod-like shaped SiO2 with sizes The mechanical properties of the scaffolds in similar to those of bone minerals. The SiO2 NPs terms of tensile strength of the CS/SiO2 scaffolds prepared at that preferable condition, possessing were determined using a Zwick Tensiler Z 2.5 small sizes (a length of 113±17 nm and a width of testing machine at a crosshead speed of 1 mm/min. 49±7.7 nm) were further used to fabricate 3D Before measurement, the scaffold specimens were CS/SiO2 composite scaffolds. Previous studies have made in the form of thin films, which were then cut shown that inorganic nanocrystals with an average into a dumbbell shape with sizes following the diameter of 40 nm and a length of 125 nm facilitated ASTM D882 standard. the rapid formation of a bone-like mineral layer on a polymer film that helps cells interact with biological 3. RESULTS AND DISCUSSION materials.[13-15] Therefore, our results indicated that the SiO2 obtained at the preferable condition is a 3.1. Characterization of the SiO2 NPs by SEM, potential inorganic material used for the fabrication XRD, and FTIR of BTE scaffolds. The typical XRD pattern of SiO2 NPs (figure 2a) Representative FE-SEM images of the nano-SiO2 was characterized by a broad peak at 2θ = 22.7o, samples synthesized by hydrothermal treatment with which revealed the presence of SiO2 as an the aid of different amounts of CTAB are presented amorphous phase on the synthesized samples. In in figure 1. The results show that the SiO2 NPs addition, no characteristic peak of other phases was synthesized by adding 1.3 g of CTAB (figure 1a) found in the XRD pattern, which could confirm that have a uniform and rod-like shape. The average a pure SiO2 phase was produced for the synthesized length and width of the SiO2 NPs were 113 and 49 scaffolds. The SiO2 NPs were further characterized nm, respectively. The dimensions of those SiO2 NPs by FTIR analysis. Figure 3a represents the FTIR are smaller than that of the SiO2 NPs reported in our spectrum of the representative SiO2 NPs, prepared at previous study by adding different amounts of the preferable condition (with adding 1.3 g of CTAB). As shown in figure 3a, the absorption Figure 1: SEM images of the SiO2 NPs synthesized by hydrothermal method with adding different amounts of CTAB: (a) 1.3 g CTAB and (b) 1.45 g CTAB, with a magnification of 50,000 © 2023 Vietnam Academy of Science and Technology, Hanoi & Wiley-VCH GmbH www.vjc.wiley-vch.de 607
25728288, 2023, 5, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/vjch.202300020 by Readcube (Labtiva Inc.), Wiley Online Library on [01/05/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Vietnam Journal of Chemistry Nguyen Kim Nga et al. the composite scaffolds. The synthesized composite scaffolds show stable shapes, a thickness of about 2 mm, and porous structures. A digital camera image of the typical composite scaffold (CS/Si1) was presented in figure 2d. The CS/SiO2 scaffolds were first characterized by XRD method. The representative XRD pattern of the CS/SiO2 scaffolds is shown in figure 2c. For comparison, the XRD pattern of bulk CS was shown in figure 2b, indicating a typical broad peak at about 2θ = 19.98o, characterized by the amorphous CS phase. A broad peak at about 2θ = 20o was observed in the XRD pattern of the CS/SiO2, which indicated the presence of CS as an amorphous and main phase in the scaffold that overlapped with that of the amorphous Figure 2: XRD patterns of (a) SiO2 NPs, (b) bulk CS phase of the SiO2 NPs. (for comparison), (c) 3D CS/SiO2 composite The presence of CS and SiO2 in the scaffolds scaffolds, (d) Digital camera image of a was further analyzed through FTIR analyses. In the representative 3D CS/SiO2 scaffold spectrum of the CS/SiO2 scaffold (figure 3c), the characteristic band at 3287.14 cm-1 was attributed to bands observed at 3453.91 and 1634.88 cm-1 can be the N-H stretching vibration of the NH2 group, assigned to the stretching and bending vibrations of whereas the bands at 1651.12 and 1557.83 cm-1 were the O-H bond of adsorbed water, respectively. The assigned to the C=O stretching vibration of amide small band at 1384.38 cm-1 was attributed to the group and the N-H bending vibration of the NH2 vibration of C-H bonds from the remnant of the group, respectively. The band at 2874.86 cm-1 was surfactant. The bands at 1098.79 cm-1 and 469.38 cm-1 were characteristic of asymmetric Si-O-Si the stretching vibration of the C-H bond of −CH2 stretching and bending vibrations, respectively. and −CH3 groups, while the bands at 1374.54 and FTIR results show that the synthesized material 1314.56 cm-1 were characteristic of the bending exists in the form of pure silica with some adsorbed vibration of the C-H bond and pyranose ring, water. respectively. The band at 1150.72 cm-1 indicated the stretching vibration of the C-O-C linkage[16], and the small band at 895.46 cm-1 was attributed to the vibrations of the saccharide structure of CS.[16] The FTIR spectrum of CS (figure 3b, for comparison) exhibited all characteristic bands observed for functional groups of CS at 3377.86, 2895.55, 1646.19, 1557.02, 1374.12, 1318.00, 1159.12, and 897.92 cm-1. The band at 3377.86 cm-1 was attributed to the stretching N-H vibration of –NH2 groups, which overlapped with the stretching vibration of hydroxyl (–OH) groups. The band observed at 2895.55 cm-1 was attributed to the stretching vibration of the C–H bond of –CH2, –CH3 groups, whereas the bands at 1374.12 and 1318.00 cm-1 are the bending vibrations of the C-H bond and pyranose ring. The bands at 1646.19 and 1557.02 Figure 3: FT-IR spectra of (a) SiO2 NPs, (b) CS, and cm-1 were characteristic of the C=O stretching (c) CS/SiO2 scaffolds vibration of the amide group and the bending vibration of the –NH2 group, respectively. The bands 3.2. Characterizations of 3D CS/SiO2 scaffolds at 1159.12 and 897.92 cm-1 are the stretching vibrations of the C-O-C linkage and vibrations of the CS/SiO2 composite scaffolds were synthesized with saccharide structure of CS. Typical bands, which are two different amounts of NaCl of 16 and 17 g responsible for stretching (1025.98 and 597.88 cm-1) (CS/Si1 and CS/Si2, respectively). Two CS and bending vibrations (555.63 and 547.92 cm-1) of scaffolds were also synthesized for comparison with Si-O bonds can be observed at, respectively, for the © 2023 Vietnam Academy of Science and Technology, Hanoi & Wiley-VCH GmbH www.vjc.wiley-vch.de 608
25728288, 2023, 5, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/vjch.202300020 by Readcube (Labtiva Inc.), Wiley Online Library on [01/05/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Vietnam Journal of Chemistry Solvent casting-particulate leaching synthesis of… CS/SiO2 scaffold (figure 3c).[17] The bands of Si-O pure CS indicating that some molecular interactions vibrations and all bands characteristic of CS between SiO2 NPs and CS in the composite functional groups in the composite scaffold showed scaffolds may have occurred, thus making SiO2 NPs small deviations compared to those of SiO2 NPs and stably dispersed in the CS matrix as a substrate. Figure 4: SEM images of the scaffolds synthesized at different amounts of NaCl: (a) CS1 and (b) CS/Si1 with 16.2 g of NaCl, (c) CS2 and (d) CS/Si2 with 17.2 g NaCl with a magnification of 50 and Pore width distributions of (e) CS/Si1 scaffold and (f) CS/Si2 scaffold The surface morphology and pore structure of average pores of 332 µm (CS2 scaffolds), 269 µm the scaffolds were examined by SEM observations. (CS/Si2). Moreover, the pore width distributions of SEM images of CS and CS/SiO2 scaffolds in figure the CS/SiO2 composite scaffolds are shown in 4 shows that all scaffolds have very porous figures 4e and f, indicating that disordered pore sizes structures. The pore sizes of the synthesized were observed for those scaffolds. It also can be scaffolds: CS1 (figure 4a), CS/Si1 (figure 4b), CS2 seen that the surface morphology of CS scaffolds (figure 4c), and CS/Si2 (figure 4d) determined was smooth (figure 5a), whereas coarse surface through ImageJ software are 262±87 µm, 198± morphology was produced for CS/SiO2 scaffolds 54µm, 332±104 µm, and 269±86 µm, respectively. (figure 5b). The observations indicated that SiO2 The results show that the amount of the porogen NPs were successfully deposited within the pore agent significantly affects the pore sizes of the walls of the scaffolds. This layer of SiO2 may help scaffolds. The use of 16 g of NaCl resulted in the cells adhere easier and interact better with CS/SiO2 formation of micropore networks with average sizes scaffolds than with CS scaffolds. SiO2 content of 262 µm (CS1 scaffolds), 198 µm (CS/Si1) and a greatly affects the pore size and overall morphology higher amount of NaCl of 17 g led to produce larger of the composite scaffolds. © 2023 Vietnam Academy of Science and Technology, Hanoi & Wiley-VCH GmbH www.vjc.wiley-vch.de 609
25728288, 2023, 5, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/vjch.202300020 by Readcube (Labtiva Inc.), Wiley Online Library on [01/05/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Vietnam Journal of Chemistry Nguyen Kim Nga et al. Figure 5: SEM image of typical CS and CS/SiO2 scaffolds at magnifications of 500 and 800 The porosities of the scaffolds measured by the decreasing in the scaffold’s mechanical strength liquid substitution method were relatively higher needed to allow the accommodation of a large than 70%. The mean porosity of CS/Si1 scaffolds number of cells and the maintenance of the desired was 70.99±2.88%, and this value did not structural strength. Previous studies have shown that significantly differ from the mean porosity of CS/Si2 scaffolds with moderate porosities of 60-75% were scaffolds (73.23±1.23%). Conversely, the porosities effectively used for bone growth because of the of CS scaffolds without SiO2 (79.26±2.21% for CS1 scaffold’s increased mechanical properties at those scaffold and 82.45±3.61% for CS2 scaffold) were porosities.[23,24] Scaffolds for bone tissue higher than those of the CS/SiO2 composite regeneration should have an adequate pore scaffolds. Our results showed that both the amount architecture (e.g., pore size and porosity), and of NaCl and the content of SiO2 influenced the pore suitable mechanical properties similar to those of sizes and porosity of the synthesized scaffolds. bone tissues. Therefore, the results obtained showed Further, the mechanical properties of the CS/SiO2 the synthesized CS/SiO2 scaffolds have suitable pore scaffolds were evaluated by measuring their tensile sizes, porosities, and mechanical strength, which are strength. Results showed that the tensile strengths of appropriate for bone tissue applications. CS/Si1 and CS/Si2 were 1.83 MPa and 1.57 MPa, 4. CONCLUSION respectively, which match very well with that of cancellous bone (the tensile strength of cancellous In this work, SiO2 NPs having a uniform-rod shape bone is ranging from 1 to 5 MPa[5,18]). with sizes similar to those of bone minerals (a length It is known that the diverse nature of tissue of 113±17 nm and a width of 49±7.7 nm) were architecture requires different micro-environments obtained by the hydrothermal method with the aid of for regeneration, including the employment of 1.3 g of CTAB. 3D biomimetic CS/SiO2 scaffolds scaffolds with preferable pore sizes.[7] In addition, a were successfully synthesized by solvent casting good distribution of the required large number of combined with a salt-leaching technique with the use cells needs high internal surface area to volume of SiO2 NPs as an inorganic component. The ratios.[19] It was indicated that typically, a pore size CS/SiO2 scaffolds exhibited suitable average pore in the range of 200-400 m is required to facilitate sizes and porosities ranging from 198 µm to 269 µm new bone formation and vascularization.[20,21] When and 70.99% to 73.23%, respectively. The tensile pore size is too small (less than 50 m), cells may strengths of the CS/SiO2 scaffolds match well with cause pore occlusion and prevent cell penetration that of cancellous bone. The results confirmed that within the scaffold, because the size of the CS/SiO2 scaffolds could support cell migration, mammalian cells is 10-30 m; pore size ranges from growth, and distribution when used as bone 75-100 m resulted in ingrowth of unmineralized scaffolds. Therefore, the results obtained osteoid tissue and smaller pores (down to 10 m) demonstrated that the synthesized CS/SiO2 scaffolds were penetrated only by fibrous tissue.[22] Porosity is should be potential biomaterials for BTE another important factor for BTE scaffolds. A high applications. porosity (higher than 90%) could provide large pore volume and internal surface area for cell adhesion, Acknowledgment. This study was funded by the proliferation, and reorganization and this would Vietnam National Foundation for Science and provide the necessary space for neovascularization Technology Development (NAFOSTED) under grant in vivo. However, an increase in porosity results in number 104.03-2019.313. © 2023 Vietnam Academy of Science and Technology, Hanoi & Wiley-VCH GmbH www.vjc.wiley-vch.de 610
25728288, 2023, 5, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/vjch.202300020 by Readcube (Labtiva Inc.), Wiley Online Library on [01/05/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Vietnam Journal of Chemistry Solvent casting-particulate leaching synthesis of… REFERENCES applications, Vietnam J. Catal. Adsorp., 2021, 10(2), 114-118. 1. Q. Wang, J. Yan, J. Yang, B. Li. Nanomaterials 13. R. K. Chandrasekhar, M. T. Shaw, M. Wei. promise better bone repair, Mater. Today, 2016, Biodegradable HA-PLA 3-D porous scaffolds: Effect 19(8), 451-463. of nano-sized filler content on scaffold properties, 2. M. N. Collins, G. Ren, K. Young, S. Pina, R. L. Reis, Acta biomater., 2005, 1(6), 653-662. J. M. Oliveira. Scaffold fabrication technologies and 14. T. T. Hoai, N. K. Nga, L. T. Giang, T. Q. Huy, P. 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