Research Projects

Due to the periosteum's superior capacity for bone regeneration, natural periosteum transplantation has been utilised clinically to aid allografts or bionic bone scaffolds in accelerating bone healing. Its probable immunological rejection, donor-site morbidity, and limited availability, however, significantly diminish their therapeutic efficacy of isolated natural periosteum. The biomimicking design of bionic periosteum has recently gained increasing attention due to its capacity to mimic the natural extracellular microenvironment of the periosteum to promote cell proliferation, differentiation, and maturation. Our proposal aimed to test potential of a nanofibrous bionic cell sheet and 3D printed scaffold combination for periosteum and superior bone remodelling using in vitro and pre-clinical animal bone defect model. Further, fate of extra cellular matrix signalling proteins involved in the process of osteogenesis would be examined in bionic nanofibrous sheet and 3D printed scaffold. There are no studies reported on the use of 3D printed Calcium lactate gluconate (CLG) scaffold in combination with Nano-fibrous sheet using xyloglucan (seed polysaccharide) in the literature. Different percentage of CLG will be printed using fused deposition modelling printer and Nano-fibrous sheet will be developed using xyloglucan by electro spinning method. Characterization of the scaffolds will be performed using SEM, EDS, XRD, FTIR, BET, AFM to understand topography, elements presence, phase purity, surface porosity and 3D topography respectively. The tensile strength and bone mineral density of the material would be examined using Instron biomechanical testing and MicroCt respectively. In vitro studies will examine the cell viability, cytotoxicity, mineralization, proliferation of adipose stem cells and differentiation markers (Immunocytochemistry and qPCR - Osteocalcin, BMP2, Osteopontin, RUNX2, and Collagen 1 Decorin, MEPE/OF45, Osterix/Sp7, β-catenin) at different time points (0, 7, 14 and 28) days in culture. A battery of extra cellular matrix proteins (Type I collagen, DEcorin, Biglycan, MGP, R-spondin, DME/MEPE) expressed during osteogenic differentiation of adipose cells will be examined by optical sectioning and western blotting method. Further, critical size femur bone defect model would be created and post-implanted with 3D printed scaffold and Nano-fibrous cell sheet as periosteum. X-ray (4 and 12th week) microct (12th week), histopathology and immunohistochemistry (Osteocalcin, BMP2, Osteopontin, Runx2, Collagen 1 Decorin, MEPE/OF45, Osterix/Sp7, β-catenin) will be performed to assess the regenerated periosteum and bone mineralization index. Complete blood count monitored. The outcome of the study will lead to the large scale inhouse bio-fabrication of a novel 3D scaffold/Nanofibrous bionic cell sheet and insights of ECM microenvironment eventually beneficial for treating critical size bone defect in patients at faster phase in near future.