Skip to main content Accessibility help

Alginate-honey bioinks with improved cell responses for applications as bioprinted tissue engineered constructs

  • Sudipto Datta (a1), Ripon Sarkar (a1), Veena Vyas (a1), Sumant Bhutoria (a2), Ananya Barui (a1), Amit Roy Chowdhury (a3) and Pallab Datta (a1)...


The polysaccharide alginate has received most extensive attention as bioink in bioprinting applications due to its ability to undergo gelation under cell-friendly conditions. However, absence of cell-binding motifs and the erratic degradation of alginate hydrogels have remained their persistent limitations. Honey is a conveniently available natural material, known for its role in wound healing and skin tissue regeneration. However, honey blending to improve biological response of alginate-based bioprinted scaffolds has not been yet reported. In the present work, honey-alginate bioinks were evaluated for their printability property (shape fidelity). It was found that honey blending reduced alginate viscosity, which gradually affected bioprinting fidelity. Therefore, the concentration that provides for acceptable bioprinting along with improvement in cell proliferations is determined. It is concluded that honey blending improves cell response of alginate bioinks and can be a facile approach to obtain bioinks especially for in situ skin tissue engineering applications.


Corresponding author

a)Address all correspondence to this author. e-mail:,


Hide All
1.Bose, S., Ke, D., Sahasrabudhe, H., and Bandyopadhyay, A.: Additive manufacturing of biomaterials. Prog. Mater. Sci. 93, 45 (2018).
2.Ozbolat, I.T., Pemg, W., and Ozbolat, V.: Application areas of 3D bioprinting. Drug Discov. Today 21, 1257 (2017).
3.Egan, P.F., Gonella, V.C., Engensperger, M., Ferguson, S.J., and Shea, K.: Computationally designed lattices with tuned properties for tissue engineering using 3D printing. PLoS One 12, e0182902 (2017).
4.Peng, W., Datta, P., Ayan, B., Ozbolat, V., Sosnoski, D., and Ozbolat, I.T.: 3D bioprinting for drug discovery and development in pharmaceutics. Acta Biomater. 57, 26 (2017).
5.Ning, L. and Chen, X.: A brief review of extrusion-based tissue scaffold bio-printing. Biotechnol. J. 12, 1600671 (2017).
6.Ozbolat, I.T. and Hospodiuk, M.: Current advances and future perspectives in extrusion-based bioprinting. Biomaterials 76, 321 (2016).
7.Cubo, N., Garcia, M., Cañizo, J.F., Velasco, D., and Jorcano, J.L.: 3D bioprinting of functional human skin: Production and in vivo analysis. Biofabrication 9 (2016).
8.Duchi, S., Onofrillo, C., Connell, C.D.O., Blanchard, R., Quigle, A.F., Kapsa, R.M.I., Peter, P., Wallace, G., Di Bella, C., and Choong, P.F.M.: Handheld co-axial bioprinting: Application to in situ surgical cartilage repair. Sci. Rep. 7, 5837 (2017).
9.Binder, K.W., Zhao, W., Aboushwareb, T., Dice, D., Atala, A., and Yoo, J.J.: In situ bioprinting of the skin for burns. J. Am. Coll. Surg. 211, S76 (2010).
10.Hospodiuk, M., Dey, M., Sosnoski, D., and Ozbolat, I.T.: The bioink: A comprehensive review on bioprintable materials. Biotechnol. Adv. 35, 217 (2017).
11.Williams, S.K. and Hoying, J.B.: Bioinks for bioprinting. In Bioprinting in Regenerative Medicine, Turksen, K., ed. (Springer International Publishing, Cham, 2015); pp. 131.
12.Hölzl, K., Lin, S., Tytgat, L., Van Vlierberghe, S., Gu, L., and Ovsianikov, A.: Bioink properties before, during and after 3D bioprinting. Biofabrication 8, 032002 (2016).
13.Abbadessa, A., Blokzijl, M.M., Mouser, V.H.M., Marica, P., Malda, J., Hennink, W.E., and Vermonden, T.: A thermo-responsive and photo-polymerizable chondroitin sulfate-based hydrogel for 3D printing applications. Carbohydr. Polym. 149, 163 (2016).
14.Billiet, T., Vandenhaute, M., Schelfhout, J., Vlierberghe, S., and Dubruel, P.: A review of trends and limitations in hydrogel-rapid prototyping for tissue engineering. Biomaterials 33, 60206041 (2012).
15.Wu, Z., Su, X., Xu, Y., Kong, B., Sun, W., and Mi, S.: Bioprinting three-dimensional cell-laden tissue constructs with controllable degradation. Sci. Rep. 6, 24474 (2016).
16.Jia, J., Richards, D.J., Pollard, S., Tan, Y., Rodriguez, J., Visconti, R.P., Trusk, T.C., Yost, M.J., Yao, H., Markwald, R.R., and Mei, Y.: Engineering alginate as bioink for bioprinting. Acta Biomater. 10, 4323 (2014).
17.Luo, Y., Luo, G., Gelinsky, M., Huang, P., and Ruan, C.: 3D bioprinting scaffold using alginate/polyvinyl alcohol bioinks. Mater. Lett. 189, 295 (2017).
18.Wan, L.Q., Jiang, J., Arnold, D.E., Guo, X.E., Lu, H.H., and Mow, V.C.: Calcium concentration effects on the mechanical and biochemical properties of chondrocyte-alginate constructs. Cell. Mol. Bioeng. 1, 93 (2008).
19.Vandamme, L., Heyneman, A., Hoeksema, H., Verbelen, J., and Monstrey, S.: Honey in modern wound care: A systematic review. Burns 39, 1514 (2013).
20.Saikaly, S.K. and Khachemoune, A.: Honey and wound healing: An update. Am. J. Clin. Dermatol. 18, 237 (2017).
21.El-kased, R.F., Amer, R.I., Attia, D., and Elmazar, M.M.: Honey-based hydrogel: In vitro and comparative in vivo evaluation for burn wound healing. Sci. Rep. 7, 9692 (2017).
22.Choi, D.S., Kim, S., Lim, Y., Gwon, H., Park, J.S., Nho, Y., Kwon, J., and Honey, C.: Hydrogel incorporated with chestnut honey accelerates wound healing and promotes early HO-1 protein expression in diabetic (db/db) mice. Tissue Eng. Regener. Med. 9, 36 (2012).
23.Majtan, J.: Honey: An immunomodulator in wound healing. Wound Repair Regen. 22, 187 (2014).
24.Wang, T., Zhu, X.K., Xue, X.T., and Wu, D.Y.: Hydrogel sheets of chitosan, honey and gelatin as burn wound dressings. Carbohydr. Polym. 88, 75 (2012).
25.Sarhan, W.A. and Azzazy, H.M.E.: High concentration honey chitosan electrospun nanofibers: Biocompatibility and antibacterial effects. Carbohydr. Polym. 122, 135 (2015).
26.Tavakoli, J. and Tang, Y.: Honey/PVA hybrid wound dressings with controlled release of antibiotics: Structural, physico-mechanical and in-vitro biomedical studies. Mater. Sci. Eng., C 77, 318 (2017).
27.Chaudhury, A., Bag, S., Barui, A., Banerjee, P., and Chatterjee, J.: Honey dilution impact on in vitro wound healing: Normoxic and hypoxic condition. Wound Repair Regen. 23, 412 (2015).
28.Wang, L., Xu, M., Luo, L., Zhou, Y., and Si, P.: Iterative feedback bio-printing-derived cell-laden hydrogel scaffolds with optimal geometrical fidelity and cellular controllability. Sci. Rep. 8, 2802 (2018).
29.Ribeiro, A., Blokzijl, M.M., Levato, R., Visser, C.W., Castilho, M., Hennink, W.E., Vermonden, T., and Malda, J.: Assessing bioink shape fidelity to aid material development in 3D bioprinting. Biofabrication 10, 014102 (2017).
30.Ouyang, L., Highley, C.B., Rodell, C.B., Sun, W., and Burdick, J.A.: 3D printing of shear-thinning hyaluronic acid hydrogels with secondary cross-linking. ACS Biomater. Sci. Eng. 2, 1743 (2016).
31.Ouyang, L., Yao, R., Zhao, Y., and Sun, W.: Effect of bioink properties on printability and cell viability for 3D bioplotting of embryonic stem cells. Biofabrication 8, 035020 (2016).
32.Kyle, S., Jessop, Z.M., Al-sabah, A., and Whitaker, I.S.: “Printability” of candidate biomaterials for extrusion based 3D printing: State-of-the-art. Adv. Healthcare Mater. 6, 1700264 (2017).
33.Di Giuseppe, M., Law, N., Webb, B., Macrae, R.A., Liew, L.J., Sercombe, T.B., Dilley, R.J., and Doyle, B.J.: Mechanical behaviour of alginate-gelatin hydrogels for 3D bioprinting. J. Mech. Behav. Biomed. Mater. 79, 150 (2018).
34.Hixon, K.R., Lu, T., Carletta, M.N., McBride-Gagyi, S.H., Janowiak, B.E., and Sell, S.A.: A preliminary in vitro evaluation of the bioactive potential of cryogel scaffolds incorporated with Manuka honey for the treatment of chronic bone infections. J. Biomed. Mater. Res., Part B 106, 19181933 (2017).
35.Sarkar, R., Ghosh, A., Barui, A., and Datta, P.: Repositing honey incorporated electrospun nanofiber membranes to provide anti-oxidant, anti-bacterial and anti-inflammatory microenvironment for wound regeneration. J. Mater. Sci.: Mater. Med. 29, 31 (2018).
36.He, Y., Yang, F., Zhao, H., Gao, Q., Xia, B., and Fu, J.: Research on the printability of hydrogels in 3D bioprinting. Sci. Rep. 6, 1 (2016).
37.Schmitt, A., Rödel, P., Anamur, C., Seeliger, C., Imhoff, A.B., Herbst, E., Vogt, S., Van Griensven, M., Winter, G., and Engert, J.: Calcium alginate gels as stem cell matrix-making paracrine stem cell activity available for enhanced healing after surgery. PLoS One 10, 1 (2015).
38.Larsen, B.E., Bjørnstad, J., Pettersen, E.O., Tønnesen, H.H., and Melvik, J.E.: Rheological characterization of an injectable alginate gel system. BMC Biotechnol. 15, 1 (2015).
39.Chung, J.H.Y., Naficy, S., Yue, Z., Kapsa, R., Quigley, A., Moulton, S.E., and Wallace, G.G.: Bio-ink properties and printability for extrusion printing living cells. Biomater. Sci. 1, 763 (2013).
40.Park, J., Lee, S.J., Chung, S., Lee, J.H., Kim, W.D., Lee, J.Y., and Park, S.A.: Cell-laden 3D bioprinting hydrogel matrix depending on different compositions for soft tissue engineering: Characterization and evaluation. Mater. Sci. Eng., C 71, 678 (2017).
41.Di Giuseppe, M., Law, N., Webb, B., Macrae, R.A., Sercombe, T.B., Dilley, R.J., Doyle, B.J., and Liew, L.J.: Journal of the mechanical behavior of biomedical materials mechanical behaviour of alginate-gelatin hydrogels for 3D bioprinting. J. Mech. Behav. Biomed. Mater. 79, 150 (2018).
42.Kaklamani, G., Cheneler, D., Grover, L.M., Adams, M.J., and Bowen, J.: Mechanical properties of alginate hydrogels manufactured using external gelation. J. Mech. Behav. Biomed. Mater. 36, 135 (2014).
43.Ahearne, M., Yang, Y., and Liu, K.: Mechanical characterisation of hydrogels for tissue engineering applications. Tissue Eng. 4, 1 (2008).
44.Hixon, K.R., Lu, T., McBride-Gagyi, S.H., Janowiak, B.E., and Sell, S.A.: A comparison of tissue engineering scaffolds incorporated with manuka honey of varying UMF. BioMed Res. Int. 2017, 4843065 (2017).
45.Minden-Birkenmaier, B.A., Neuhalfen, R.M., Janowiak, B.E., and Sell, S.: Preliminary investigation and characterization of electrospun polycaprolactone and manuka honey scaffolds for dermal repair. J. Eng. Fibers Fabr. 10, 126 (2015).
46.Rajput, M., Bhandaru, N., Barui, A., Chaudhary, A., Paul, R.R., Mukherjee, R., and Chatterjee, J.: Nano-patterned honey incorporated silk fibroin membranes for improving cellular compatibility. RSC Adv. 4, 44674 (2014).
47.Funada, M., Hara, H., Sasagawa, H., Kitagawa, Y., and Kadowaki, T.: A honey bee Dscam family member, AbsCAM, is a brain-specific cell adhesion molecule with the neurite outgrowth activity which influences neuronal wiring during development. Eur. J. Neurosci. 25, 168 (2007).
48.Nordin, A., Sainik, N.Q.A.V., Zulfarina, M.S., Naina-Mohamed, I., Saim, A., and Bt Hj Idrus, R.: Honey and epithelial to mesenchymal transition in wound healing: An evidence-based review. Wound Med. 18, 8 (2017).
49.Oryan, A., Alemzadeh, E., and Moshiri, A.: Biological properties and therapeutic activities of honey in wound healing: A narrative review and meta-analysis. J. Tissue Viability 25, 98 (2016).
50.Daly, A.C., Freeman, F.E., Gonzalez-Fernandez, T., Critchley, S.E., Nulty, J., and Kelly, D.J.: 3D bioprinting for cartilage and osteochondral tissue engineering. Adv. Healthcare Mater. 6, 1700298 (2017).
51.Nair, K., Gandhi, M., Khalil, S., Yan, K.C., Marcolongo, M., Barbee, K., and Sun, W.: Characterization of cell viability during bioprinting processes. Biotechnol. J. 4, 1168 (2009).
52.He, P., Zhao, J., Zhang, J., Li, B., Gou, Z., Gou, M., and Li, X.: Bioprinting of skin constructs for wound healing. Burn. Trauma 6, 5 (2018).
53.Barui, A., Banerjee, P., Chaudhary, A., Conjeti, S., Mondal, B., Dey, S., and Chatterjee, J.: Evaluation of angiogenesis in diabetic lower limb wound healing using a natural medicine: A quantitative approach. Wound Med. 6, 26 (2014).



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed