title: Cross-correlative 3D micro-structural investigation of human bone processed into bone allografts.
abstract: Bone allografts (BA) are a cost-effective and sustainable alternative in orthopedic practice as they provide a permanent solution for preserving skeletal architecture and function. Such BA however, must be processed to be disease free and immunologically safe as well as biologically and clinically useful. Here, we have demonstrated a processing protocol for bone allografts and investigated the micro-structural properties of bone collected from osteoporotic and normal human donor samples. In order to characterize BA at different microscopic levels, a combination of techniques such as Solid State Nuclear Magnetic Resonance (ssNMR), Scanning Electron Microscope (SEM), micro-computed tomography (μCT) and Thermal Gravimetric Analysis (TGA) were used for delineating the ultra-structural property of bone. ssNMR revealed the extent of water, collagen fine structure and crystalline order in the bone. These were greatly perturbed in the bone taken from osteoporotic bone donor. Among the processing methods analyzed, pasteurization at 60 °C and radiation treatment appeared to substantially alter the bone integrity. SEM study showed a reduction in Ca/P ratio and non-uniform distribution of elements in osteoporotic bones. μ-CT and MIMICS (Materialize Interactive Medical Image Control System) demonstrated that pasteurization and radiation treatment affects the BA morphology and cause a shift in the HU unit. However, the combination of all these processes restored all-important parameters that are critical for BA integrity and sustainability. Cross-correlation between the various probes we used quantitatively demonstrated differences in morphological and micro-structural properties between BA taken from normal and osteoporotic human donor. Such details could also be instrumental in designing an appropriate bone scaffold. For the best restoration of bone microstructure and to be used as a biomaterial allograft, a step-wise processing method is recommended that preserves all critical parameters of bone, showing a significant advancements over currently existing methods.
authors.affiliation: Centre for Research in Nanotechnology & Science, Indian Institute of Technology Bombay, Mumbai 400076, India.
authors.forename: Atul Kumar
authors.initials: AK
authors.lastname: Singh
authors.affiliation: Tissue Bank, Tata Memorial Hospital, Parel, Mumbai 400012, India.
authors.forename: Astrid Lobo
authors.initials: AL
authors.lastname: Gajiwala
authors.affiliation: Centre of Biomedical Research, SGPGIMS Campus, Lucknow 226014, India.
authors.forename: Ratan Kumar
authors.initials: RK
authors.lastname: Rai
authors.affiliation: Division of Endocrinology, Center for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI) CSIR-Central Drug Research Institute, Lucknow 226031, India.
authors.forename: Mohd Parvez
authors.initials: MP
authors.lastname: Khan
authors.affiliation: Centre of Biomedical Research, SGPGIMS Campus, Lucknow 226014, India.
authors.forename: Chandan
authors.initials: C
authors.lastname: Singh
authors.affiliation: Division of Endocrinology, Center for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI) CSIR-Central Drug Research Institute, Lucknow 226031, India.
authors.forename: Tarun
authors.initials: T
authors.lastname: Barbhuyan
authors.affiliation: Centre for Research in Nanotechnology & Science, Indian Institute of Technology Bombay, Mumbai 400076, India.
authors.forename: S
authors.initials: S
authors.lastname: Vijayalakshmi
authors.affiliation: Division of Endocrinology, Center for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI) CSIR-Central Drug Research Institute, Lucknow 226031, India.
authors.forename: Naibedya
authors.initials: N
authors.lastname: Chattopadhyay
authors.affiliation: Centre of Biomedical Research, SGPGIMS Campus, Lucknow 226014, India. Electronic address: neerajcbmr@gmail.com.
authors.forename: Neeraj
authors.initials: N
authors.lastname: Sinha
authors.affiliation: Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400076, India. Electronic address: ashutoshk@iitb.ac.in.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Centre for Research in Nanotechnology & Science, Indian Institute of Technology Bombay, Mumbai 400076, India; Department of Chemical Engineering, Indian Institute of Technology Bombay, Mumbai 400076, India. Electronic address: jb@iitb.ac.in.
authors.forename: Jayesh R
authors.initials: JR
authors.lastname: Bellare
mesh_terms: D016025:Bone Transplantation; D001842:Bone and Bones / Q000000981:diagnostic imaging*; D003094:Collagen / Q000737:chemistry; D006801:Humans; D008279:Magnetic Resonance Imaging; D009682:Magnetic Resonance Spectroscopy; D008855:Microscopy, Electron, Scanning; D013818:Thermogravimetry; D014184:Transplantation, Homologous; D014867:Water / Q000737:chemistry; D055114:X-Ray Microtomography
chemical_list: D014867:Water; D003094:Collagen
keywords: Allograft; Bone; Human bone; MIMICS; Processing; Scanning electron microscopy; Solid state NMR