Development of PLA/HA porous scaffolds with controlled pore sizes using the combined freeze drying and sucrose leaching technique for bone tissue engineering


  • Sunisa SINGHAWANNURAT School of Chemical Engineering, Institute of Engineering, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand
  • Panuwat LAWTAE School of Chemical Engineering, Institute of Engineering, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand
  • Catleya ROJVIRIYA Synchrotron Light Research Institute (Public Organization), Nakhon Ratchasima, 30000, Thailand
  • Chalermluck PHOOVASAWAT Synchrotron Light Research Institute (Public Organization), Nakhon Ratchasima, 30000, Thailand



porous scaffold, Hydroxyapatite, poly-lactic acid, a combined freeze drying/sucrose leaching technique, porosity


The combination of freeze drying and sucrose leaching technique was employed to fabricate PLA/HA scaffolds with controlled pore size. The influence of the HA content and sucrose size on the scaffold properties was investigated. The fabricated scaffolds showed porous properties with a porosity of 44% to 58% and pore size of 461 μm to 688 μm. The results indicated that the scaffolds possessed favorable porous properties, illustrated by good interconnectivity, appropriate pore size, and suitable porosity. These characteristics were crucial for facilitating bone cell growth and promoting the formation of new tissue within the scaffold structure. The compressive modulus of the scaffolds was examined and found to be in the range of 3.35 MPa to 5.75 MPa. Furthermore, the degradation behavior of the scaffolds was studied for 28 days in a Phosphate Buffered Saline solution. The results showed that the degradation rate was varied in the range of 6% to 14%. The water uptake of the scaffolds exhibited a range between 180% and 200%. Enhancement in water uptake was observed with higher HA content and increased sucrose size. Consequently, the scaffolds developed in this study hold promise as optimal candidates for bone tissue engineering applications.


Download data is not yet available.


R. Dimitriou, E. Jones, D. McGonagle, and P. V. Giannoudis, "Bone regeneration: current concepts and future directions," BMC Medicine, vol. 9, no. 1, p. 66, 2011.

C. Kiernan, C. Knuth, and E. Farrell, "Chapter 6 - endochondral ossification: recapitulating bone development for bone defect repair," in Developmental Biology and Musculoskeletal Tissue Engineering, M. J. Stoddart, A. M. Craft, G. Pattappa, and O. F. W. Gardner Eds. Boston: Academic Press, pp. 125-148, 2018.

A. Ibrahim, "13 - 3D bioprinting bone," in 3D Bioprinting for Reconstructive Surgery, D. J. Thomas, Z. M. Jessop, and I. S. Whitaker Eds.: Woodhead Publishing, pp. 245-275, 2018.

M. S. Carvalho, J. Cabral, C. Silva, and D. Vashishth, "Bone matrix non-collagenous proteins in tissue engineering: creating new bone by mimicking the extracellular matrix," Polymers, vol. 13, p. 1095, 2021.

S. Pramanik, S. Kharche, N. More, D. Ranglani, G. Singh, and G. Kapusetti, "Natural biopolymers for bone tissue engineering: A brief review," Engineered Regeneration, vol. 4, no. 2, pp. 193-204, 2023.

B. N. Kharbikar, J. X. Zhong, D. L. Cuylear, C. A. Perez, and T. A. Desai, "Theranostic biomaterials for tissue engineering," Current Opinion in Biomedical Engineering, vol. 19, p. 100299, 2021.

H. Qu, H. Fu, Z. Han, and Y. Sun, "Biomaterials for bone tissue engineering scaffolds: A review," (in Eng), RSC Advances, vol. 9, no. 45, pp. 26252-26262, 2019.

D. T. Dixon, and C. T. Gomillion, "Conductive scaffolds for bone tissue engineering: Current state and future outlook," (in Eng), Journal of Functional Biomaterials, vol. 13, no. 1, 2021.

M. Ferraz, F. Monteiro, and C. Manuel, "Hydroxyapatite nano-particles: A review of preparation methodologies," Journal of applied biomaterials & biomechanics: Journal of Applied Biology and Biotechnology, vol. 2, pp. 74-80, 2003.

M. Eilbagi, R. Emadi, K. Raeissi, M. Kharaziha, and A. Valiani, "Mechanical and cytotoxicity evaluation of nanostructured hydroxyapatite-bredigite scaffolds for bone regeneration," Materials Science and Engineering: C, vol. 68, pp. 603-612, 2016.

K. Aoki, and N. Saito, "Biodegradable polymers as drug delivery systems for bone regeneration," (in Eng), Pharmaceutics, vol. 12, no. 2, 2020.

S. Hassanajili, A. Karami-Pour, A. Oryan, and T. Talaei-Khozani, "Preparation and characterization of PLA/PCL/HA composite scaffolds using indirect 3D printing for bone tissue engineering," Materials Science and Engineering: C, vol. 104, p. 109960, 2019.

F. Donnaloja, E. Jacchetti, M. Soncini, and M. T. Raimondi, "Natural and synthetic polymers for bone scaffolds optimization," (in Eng), Polymers (Basel), vol. 12, no. 4, 2020.

S. Bhushan, S. Singh, T. K. Maiti, C. Sharma, D. Dutt, S. Sharma, C. H. Li, E. M. T. Eldin, "Scaffold fabrication techniques of biomaterials for bone tissue engineering: A critical review," Bioengineering, vol. 9, no. 12, 2022.

J. Xing, M. Zhang, X. Liu, C. Wang, N. Xu, and D. Xing, "Multi-material electrospinning: from methods to biomedical applications," Materials Today Bio, vol. 21, p. 100710, 2023.

C. Wang, W. Huang, Y. Zhou, L. He, Z. He, Z. Chen, X, He, S. Tian, J. Liao, B. Lu, Y. Wei, and M. Wang, "3D printing of bone tissue engineering scaffolds," Bioactive Materials, vol. 5, no. 1, pp. 82-91, 2020.

H. J. Park, O. J. Lee, M. C. Lee, B. M. Moon, H. W. Ju, J. M. Lee, J-H. Kim, D. W. Kim, and C. Park, "Fabrication of 3D porous silk scaffolds by particulate (salt/sucrose) leaching for bone tissue reconstruction," International Journal of Biological Macromolecules, vol. 78, pp. 215-223, 2015.

D. Mao, Q. Li, D. Li, Y. Tan, and Q. Che, "3D porous poly(ε-caprolactone)/58S bioactive glass-sodium alginate/gelatin hybrid scaffolds prepared by a modified melt molding method for bone tissue engineering," Materials & Design, vol. 160, pp. 1-8, 2018.

Y. Nam, J. Yoon, and T. Park, "A novel fabrication method for macroporous scaffolds using gas foaming salt as porogen additive," Journal of Biomedical Materials Research, vol. 53, pp. 1-7, 2000.

J. D. Chen, Y. Wang, and X. Chen, "In situ fabrication of nano-hydroxyapatite in a macroporous chitosan scaffold for tissue engineering," (in Eng), Journal of Biomaterials Science, Polymer Edition, vol. 20, no. 11, pp. 1555-65, 2009.

R. Govindan, F. L. Gu, S. Karthi, and E. K. Girija, "Effect of phosphate glass reinforcement on the mechanical and biological properties of freeze-dried gelatin composite scaffolds for bone tissue engineering applications," Materials Today Communications, vol. 22, p. 100765, 2020.

T. L. Conrad and R. K. Roeder, "Effects of porogen morphology on the architecture, permeability, and mechanical properties of hydroxyapatite whisker reinforced polyetheretherketone scaffolds," Journal of the Mechanical Behavior of Biomedical Materials, vol. 106, p. 103730, 2020.

Y. S. Cho, B.-S. Kim, H.-K. You, and Y.-S. Cho, "A novel technique for scaffold fabrication: SLUP (salt leaching using powder)," Current Applied Physics, vol. 14, no.3, pp. 371-377, 2014.

M. Alizadeh, F. Abbasi, A. B. Khoshfetrat, and H. Ghaleh, "Microstructure and characteristic properties of gelatin/chitosan scaffold prepared by a combined freeze-drying/leaching method," Materials Science and Engineering: C, vol. 33, no. 7, pp. 3958-3967, 2013.

Á. Serrano-Aroca, A. Cano-Vicent, R. S. Serra, M. El-Tanani, A. Aijabali, M. Tambuwale, and Y. K. Mishra, "Scaffolds in the microbial resistant era: Fabrication, materials, properties and tissue engineering applications," Materials Today Bio, vol. 16, p. 100412, 2022.

R. Dorati, C. Colonna, I. Genta, T. Modena, and B. Conti, "Effect of porogen on the physico-chemical properties and degradation performance of PLGA scaffolds," Polymer Degradation and Stability, vol. 95, no. 4, pp. 694-701, 2010.

H. Gheisari, E. Karamian, and M. Abdellahi, "A novel hydroxy- apatite –Hardystonite nanocomposite ceramic," Ceramics International, vol. 41, pp. 5967-5975, 2015.

E. F. Morgan, G. U. Unnikrisnan, and A. I. Hussein, "Bone mechanical properties in healthy and diseased states," (in Eng), Annual Review of Biomedical Engineering, vol. 20, pp. 119-143, 2018.

A. Chandrasekaran, S. Suresh, and A. Dakshanamoorth "Synthesis and characterization of nano-hydroxyapatite (n-HAP) using the wet chemical technique," International Journal of Physical Sciences, vol. 8, pp. 1639-1645, 2013.

M. Murugesan, D. Mangalaraj, P. Nagamony, and C. Viswanathan, "Core-shell hydroxyapatite/Mg nanostructures: Surfactant free facile synthesis, characterization and their in vitro cell viability studies against leukaemia cancer cells (K562)," RSC Advances., vol. 5, 2015.

E. Åkerlund, A. Diez-Escudero, A. Grzeszczak, and C. Persson, "The Effect of PCL Addition on 3D-Printable PLA/HA composite filaments for the treatment of bone defects," Polymers, vol. 14, no. 16, p. 3305, 2022.

B. W. Chieng, N. Ibrahim, W. Yunus, and M. Hussein, "Effects of graphene nanopletelets on poly(lactic acid)/poly (ethylene glycol) polymer nanocomposites," Polymers, vol. 6, pp. 93-104, 2013.

P. Singla, R. Mehta, D. Berek, and S. Upadhyay, "Microwave assisted synthesis of poly(lactic acid) and its characterization using size exclusion chromatography," Journal of Macro-molecular Science Part A Pure and Applied Chemistry, vol. A49, 2012.

A. Zimina, F. Senatov, R. Choudhary, E. Kolesnikov, N. Anisimova, M. Kiselevskiy, P. Orlova, N. Strukova, M. Generalova, V. Manskikh, A. Gromov, and A. Karyagina, "Biocompatibility and physico-chemical properties of highly porous PLA/HA scaffolds for bone reconstruction," (in Eng), Polymers (Basel), vol. 12, no. 12, 2020.

J. Su, J. Teng, Z. Xu, and Y. Li, "Effects of hydroxyapatite content on mechanical properties and in-vitro corrosion behavior of ZK60/HA composites," vol. 111, no. 8, pp. 621-631, 2020.

S. Ufere, and N. S. P. C. Csci, "Fabrication and characterization of PCL/HA/PPY composite scaffold using freeze-drying technique," Journal Teknologi, vol. 78, 2016.

Z. Cui, W. Li, L. Cheng, D. Gong, W. Cheng, and W. Wang, "Effect of nano-HA content on the mechanical properties, degradation and biocompatible behavior of Mg- Zn/HA composite prepared by spark plasma sintering," Materials Characterization, vol. 151, pp. 620-631, 2019.

T.-T. Li, Y. Zhang, H.-T. Ren, H.-K. Peng, C.-W. Lou, and J.-H. Lin, "Two-step strategy for constructing hierarchical pore structured chitosan–hydroxyapatite composite scaffolds for bone tissue engineering," Carbohydrate Polymers, vol. 260, p. 117765, 2021.

M. V. Chaikina, N. Bulina, O. Vinokurova, K. Gerasimov, I. Y. Prosanov, N. Kompankov, O. Lapina, E. Papulovskiv, A. Ishchenko, and S. Makarova, "Possibilities of mechano-chemical synthesis of apatites with different Ca/P ratios," Ceramics, vol. 5, no. 3, pp. 404-422, 2022.

M. R. B. M. Roslan, N. F. M. Nasir, R. Khalid, N. F. Mohammad, C. E. Meng, N. N. N. Hashim, B. C. You, M. S. A. Majid, and N. A. M. Amin, "The optimization of the hydroxy-apatite (HA) material characteristics produced from corbiculacea (Etok) shells," Journal of Physics: Conference Series, vol. 1372, p. 012077, 2019.

S. Best, B. Sim, M. Kayser, and S. Downes, "The dependence of osteoblastic response on variations in the chemical composition and physical properties of hydroxyapatite," Journal of Materials Science: Materials in Medicine, vol. 8, no. 2, pp. 97-103, 1997.

N. Aboudzadeh, A. Khavandi, J. Javadpour, M. A. Shokrgozar, and M. Imani, "Effect of dioxane and N-Methyl-2-pyrrolidone as a solvent on biocompatibility and degradation performance of PLGA/nHA scaffolds," (in Eng), Iranian Biomedical Journal, Full Length vol. 25, no. 6, pp. 408-416, 2021.

M. Haugh, C. Murphy, and F. O’Brien, "Novel freeze-drying methods to produce a range of collagen–glycosaminoglycan scaffolds with tailored mean pore sizes," Tissue engineering. Part C, Methods, vol. 16, pp. 887-94, 2009.

Y. Shafieyan, S. Sharifi, M. Imani, M. Shokrgozar, N. Aboudzadeh, and M. Atai, "A biocompatible composite based on poly(Îμ-caprolactone fumarate) and hydroxyapatite," Polymers for Advanced Technologies, vol. 22, pp. 2182-2190, 2011.

G. Huang, L. Xu, J. Wu, S. Wang, and Y. Dong, "Gelatin/ bioactive glass composite scaffold for promoting the migration and odontogenic differentiation of bone marrow mesenchymal stem cells," Polymer Testing, vol. 93, p. 106915, 2021.

R. Radakisnin, M. S. A. Majid, M. R. M. Jamir, M. F. M. Tahir, C. E. Meng, and H. A. Alshahrani, "Physical, thermal, and mechanical properties of highly porous polylactic acid/cellulose nanofibre scaffolds prepared by salt leaching technique," Nano-technology Reviews, vol. 10, no. 1, pp. 1469-1483, 2021.

Z. Shahbazarab, A. Teimouri, A. N. Chermahini, and M. Azadi, "Fabrication and characterization of nanobiocomposite scaffold of zein/chitosan/nanohydroxyapatite prepared by freeze-drying method for bone tissue engineering," International Journal of Biological Macromolecules, vol. 108, pp. 1017-1027, 2018.

P. Kazimierczak, A. Benko, K. Palka, C. Canal, D. Kolodynska, and A. Przekora, "Novel synthesis method combining a foaming agent with freeze-drying to obtain hybrid highly macroporous bone scaffolds," Journal of Materials Science & Technology, vol. 43, pp. 52-63, 2020.

S. Bose, M. Roy, and A. Bandyopadhyay, "Recent advances in bone tissue engineering scaffolds," Trends in Biotechnology, vol. 30, no. 10, pp. 546-554, 2012.




How to Cite

S. SINGHAWANNURAT, P. . LAWTAE, C. . ROJVIRIYA, and C. . PHOOVASAWAT, “Development of PLA/HA porous scaffolds with controlled pore sizes using the combined freeze drying and sucrose leaching technique for bone tissue engineering”, J Met Mater Miner, vol. 34, no. 2, p. 1928, Jun. 2024.



Original Research Articles