Fabrication of Nanocollagen Using Enhanced Cryogenic Milling Method with Graphene Oxide

Abstract

Samantha Lo,1 Ebrahim Mahmoudi,2 Looi Qi Hao,1,3 Fatimah Mohd Nor,4 Mh Busra Fauzi1,5 1Centre for Tissue Engineering Centre and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras, Kuala Lumpur, 56000, Malaysia; 2Department of Chemical and Process Engineering, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, Bangi, Selangor Darul Ehsan, 43600, Malaysia; 3My Cytohealth Sdn. Bhd, Kuala Lumpur, 56000, Malaysia; 4Plastic Surgery Department, Kumpulan Perubatan Johor Ampang Puteri Specialist Hospital, Kuala Lumpur, 68000, Malaysia; 5Advance Bioactive Materials-Cells (Adv-BioMaC) UKM Research Group, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, MalaysiaCorrespondence: Mh Busra Fauzi, Email [email protected]: Collagen, a widely used natural biomaterial polymer in skin tissue engineering, can be innovatively processed into nanocollagen through cryogenic milling to potentially enhance skin tissue healing. Although various methods for fabricating nanocollagen have been documented, there is no existing study on the fabrication of nanocollagen via cryogenic milling, specifically employing graphene oxide as separators to prevent agglomeration.Methods: In this study, three research groups were created using cryogenic milling: pure nanocollagen (Pure NC), nanocollagen with 0.005% graphene oxide (NC + 0.005% GO), and nanocollagen with 0.01% graphene oxide (NC+0.01% GO). Characterization analyses included transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, x-ray diffraction (XRD), zeta potential (ZP), and polydispersity index (PDI).Results: TEM and SEM analysis revealed that nanocollagen groups alone exhibited particle sizes of less than 100 nm. FTIR spectroscopic investigations indicated the presence of amide A, B, and I, II, and III (1800 to 800 cm− 1) in all nanocollagen study groups, with the characteristic C-O-C stretching suggesting the incorporation of graphene oxide (GO). XRD data exhibited broadening of the major peak as the proportion of GO increased from pure NC to the nanocollagen groups with GO. Zeta potential measurements indicated electrostatic attraction of the samples to negatively charged surfaces, accompanied by sample instability. PDI results depicted size diameters ranging from 800 to 1800 nm, indicating strong polydispersity with multiple size populations.Conclusion: This research demonstrated that collagen can be successfully fabricated into nanoparticles with sizes smaller than 100 nm.Keywords: nanocollagen, cryogenic milling, graphene oxide

Keywords

• nanocollagen
• cryogenic milling
• graphene oxide