Controlled pore anisotropy in chitosan-gelatin cryogels for use in bone tissue engineering

Abstract

In tissue engineering, the development of an appropriate scaffold is crucial to provide a framework for new tissue growth. The use of cryogels as scaffolds shows promise due to their macroporous structure, but the pore size, distribution, and interconnectivity is highly variable depending on the fabrication process. The objective of the current research is to provide a technique for controlled anisotropy in chitosan-gelatin cryogels to develop scaffolds for bone tissue engineering application. A mold was developed using additive manufacturing to be used during the freezing process in order to fabricate cryogels with a more interconnected pore structure. The scaffolds were tested to evaluate their porosity, mechanical strength, and to observe cell infiltration through the cryogel. It was found that the use of the mold allowed for the creation of designated pores within the cryogel structure which facilitated cell infiltration to the center of the scaffold without sacrificing mechanical integrity of the structure.

Keywords

Critical size bone defects additive manufacturing scaffold controlled porosity scaffold fabrication

Get full access to this article

View all access options for this article.

References

García

Clark

García

. Integrin specific hydrogels functionalized with VEGF for vascularization and bone regeneration of critical size bone defects. Journal of biomedical materials research Part A 2016; 104(4): 889–900.

Nauth

. Critical-size bone defects: is there a consensus for diagnosis and treatment? J Orthop Trauma 2018; 32(Suppl 1): S7–s11.

Cooper

. Testing the “critical-size” in calvarial bone defects: revisiting the concept of a critical-sized defect (CSD). Plast Reconstr Surg 2010; 125(6): 1685.

Sela

Bab

. Principles of bone regeneration. Springer Science and Business Media, 2012.

Zhang

Chen

. Bone tissue regeneration-application of mesenchymal stem cells and cellular and molecular mechanisms. Curr Stem Cell Res Ther 2017; 12(5): 357–364.

Fernandez de Grado

. Bone substitutes: a review of their characteristics, clinical use, and perspectives for large bone defects management. J Tissue Eng 2018; 9: 2041731418776819.

Myeroff

Archdeacon

. Autogenous bone graft: donor sites and techniques. JBJS 2011; 93(23): 2227–2236.

Betz

. Limitations of autograft and allograft: new synthetic solutions. Orthopedics 2002; 25(5): S561–S570.

Goldberg

. Natural history of autografts and allografts. Bone implant grafting, 1992, pp. 9–12.

10.

Dimitriou

. Bone regeneration: current concepts and future directions. BMC Med 2011; 9(1): 66.

11.

Chan

Leong

. Scaffolding in tissue engineering: general approaches and tissue-specific considerations. Eur Spine J 2008; 17(4): 467–479.

12.

Hixon

Sell

. A comprehensive review of cryogels and their roles in tissue engineering applications. Acta Biomater 2017; 62: 29–41.

13.

Hixon

. The calcification potential of cryogel scaffolds incorporated with various forms of hydroxyapatite for bone regeneration. Biomedical materials 2017; 12(2): 025005.

14.

Razavi

Qiao

Thakor

. Three dimensional cryogels for biomedical applications. J Biomed Mater Res 2019; 107(12): 2736–2755.

15.

Henderson

. Cryogels for biomedical applications. J Mater Chem B 2013; 1(21): 2682–2695.

16.

Saylan

Denizli

. Supermacroporous composite cryogels in biomedical applications. Gels (Basel, Switzerland) 2019; 5(2): 20.

17.

Kathuria

. Synthesis and characterization of elastic and macroporous chitosan–gelatin cryogels for tissue engineering. Acta Biomater 2009; 5(1): 406–418.

18.

VandeVord

. Evaluation of the biocompatibility of a chitosan scaffold in mice. J Biomed Mater Res: An Official Journal of The Society for Biomaterials, The Japanese Society for Biomaterials 2002; 59(3): 585–590.

19.

Ullm

. Biocompatibility and inflammatory response in vitro and in vivo to gelatin-based biomaterials with tailorable elastic properties. Biomaterials 2014; 35(37): 9755–9766.

20.

Abbasi

. Porous scaffolds for bone regeneration. J Sci: Advanced Materials and Devices 2020; 5(1): 1–9.

21.

Jia

. Exploring the interconnectivity of biomimetic hierarchical porous Mg scaffolds for bone tissue engineering: effects of pore size distribution on mechanical properties, degradation behavior and cell migration ability. J Magnesium Alloys 2021; 9(6): 1954–1966.

22.

Somo

. Pore interconnectivity influences growth factor-mediated vascularization in sphere-templated hydrogels. Tissue Eng C Methods 2015; 21(8): 773–785.

23.

Jones

. Quantifying the 3D macrostructure of tissue scaffolds. J Mater Sci Mater Med 2009; 20(2): 463–471.

24.

Wakim

Grewal

. Structure of Bone. Human Biology. LibreTexts 2023.

25.

Chang

Liu

. Osteon: structure, turnover, and regeneration. Tissue Engineering Part B: Reviews 2022; 28(2): 261–278.

26.

Rho

J-Y

Kuhn-Spearing

Zioupos

. Mechanical properties and the hierarchical structure of bone. Med Eng Phys 1998; 20(2): 92–102.

27.

Hixon

. A comparison of cryogel scaffolds to identify an appropriate structure for promoting bone regeneration. Biomedical Physics & Engineering Express 2016; 2(3): 035014.

28.

Hulbert

. Potential of ceramic materials as permanently implantable skeletal prostheses. J Biomed Mater Res 1970; 4(3): 433–456.

29.

Loh

Choong

. Three-dimensional scaffolds for tissue engineering applications: role of porosity and pore size. Tissue Engineering. Part B, Reviews 2013; 19(6): 485–502.

30.

Öfkeli

Demir

Bölgen

. Biomimetic mineralization of chitosan/gelatin cryogels and in vivo biocompatibility assessments for bone tissue engineering. J Appl Polym Sci 2021; 138(14): 50337.

31.

Rezwan

. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials 2006; 27(18): 3413–3431.

32.

Osterhoff

. Bone mechanical properties and changes with osteoporosis. Injury 2016; 47(Suppl 2): S11–20.

33.

Havaldar

Pilli

Putti

. Insights into the effects of tensile and compressive loadings on human femur bone, vol. 3. Advanced biomedical research, 2014.

34.

Linde

Hvid

Pongsoipetch

. Energy absorptive properties of human trabecular bone specimens during axial compression. J Orthop Res 1989; 7(3): 432–439.

35.

Borra

. A simple method to measure cell viability in proliferation and cytotoxicity assays. Braz Oral Res 2009; 23(3): 255–262.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.16 MB