Browsing by Author "Athira, RK"

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    Hydroxyapatite cages with aligned pores for bone grafting – Seeding of human osteoblast-like cells in vitro and their response in dynamic culture mode
    (Ceramics International, 2021-11) Athira, RK; Gayathry, G; Anil Kumar, PR; Harikrishna Varma, PR; Kasoju, N; Manoj, Komath
    Hydroxyapatite (HA) is a highly regarded synthetic bone graft material. Porous HA ceramics blocks are used to substitute harvested natural bone grafts. Being similar to bone mineral, HA material integrates with the host bone through surface osteointegration and slowly resorb along with the natural bone remodeling process. The blocks in use currently have random and tortuous pore structures. The present work explores the usefulness of cage-like HA ceramic design with end-to-end open pores, with the help of in vitro cell culture methods. Such a structure, on implantation, will take up the blood factors and cells and host the bone remodeling process inside the bulk of the cage, leading to early healing. In the study, HA samples with aligned through-pores were prepared and explored in vitro, with a focus on how the pores host the cells inside and to what level the cells maintain their activity. Human osteoblast-like cells (HOS) were used, at different seeding and culturing approaches. Cell seeding was done through (i) conventional large volume cell suspension, (ii) a confined mini chamber with a limited volume of cell suspension, and (iii) placing a concentrated drop of cell suspension directly on top of the scaffold. The third approach gave the best cell adhesion and proliferation, and hence used for further explorations. A dynamic culture system was designed in-house by bifurcating the cell culture wells using vertical inserts, holding the samples horizontally with their ends open to both sides, and making the media flow across using a rocker platform. The HOS cell adhesion, viability and proliferation were tested in the HA cages, in static and dynamic culture conditions, with conventional porous ceramics as the control. The cell infiltration was deeper and the cell viability over a period of 7 days was significantly higher in dynamic culture conditions in the test samples.
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    Mesenchymal stem cell culture in aligned porous hydroxyapatite scaffolds using a multiwell plate bioreactor for bone tissue engineering
    (MedComm – Future Medicine, 2022-09) Gayathry, G; Athira, RK; Anju, MS; Anil Kumar, PR; Harikrishna Varma, PR; Kasoju, N; Komath, M
    Regeneration of bone lost by trauma, diseases and aging, and restoration of its load-bearing function are major clinical challenges. Hydroxyapatite (HA) is a clinically proven scaffold material for bone grafting, but the random-pore structure limits the homing of the cells inside the graft and the bone regeneration progresses with the resorption of the graft material. This work is based on the hypothesis that aligned through pores in the graft will lead to a faster healing by homing the local cells inside and provide a better environment for new bone formation through the graft structure. The investigation was done using aligned porous HA scaffolds seeded with human Wharton's jelly-derived mesenchymal stem cells (hWJ-MSCs) and cultured in a multiwell format bioreactor setup. The cell adhesion was studied by microscopy, cell proliferation was evaluated by Alamar blue assay and osteogenic differentiation was confirmed by biochemical and molecular assays. The results indicate that the hWJ-MSCs infiltrated through the aligned porous network of the scaffold, proliferated well when cultured in the expansion medium, and differentiated into osteogenic lineage when cultured in the differentiation medium.
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    A versatile approach for temporary storage and shipping of in vitro cultured cells, cell sheets and tissue engineered constructs – a preliminary report
    (Engineered Regeneration, 2022-09) Anju, MS; Athira, RK; Ramesh Babu, V; Anil Kumar, PR; Kasoju, N
    Temporary storage/ shipping of cell/ tissue engineering products from bench to bedside is a key aspect of regenerative medicine. The current proof-of-concept study presents a multipurpose device for temporary storage/ shipping of cell culture dishes containing cell/ tissue constructs. The device, made with readily available raw materials, contains three elements viz. a specialized lid, polymeric plates having grooves and a set of nuts and bolts. As part of the performance evaluation, the device was first subjected to a simulated storage/ shipping process, wherein the leak-proof and aseptic containment of the contents was demonstrated. Subsequently, the setup was used for temporary storage/ shipping of dishes having (a) L929 cell monolayers cultured on treated surfaces, (b) SIRC, HaCaT and A549 cell sheets cultured on thermo-responsive surfaces, (c) HOS-cell encapsulated agar gels and (d) HOS-cell seeded silk fibroin mats. The results showed that the health of cell monolayers/ cell sheets/ tissue constructs after the process was comparable to that before the process. The device was scalable, simple to handle, can be made for a single or multi-use purpose, and can be resizable to load other culture vessels. The design of the storage/ shipping device described in this report thus offers versatile features and applications.