Obtaining COFRAC accreditation for the blood compatibility tests was a major target in the recent past and our team could standardize existing tests, introduce new tests, validate them, and publish the data to get recognition, simultaneously with quality system implementation. The continued demands from Industries and R&D organizations to test materials/devices for in vitro blood compatibility during the past 5 to 6 years imply that the team in TRU is offering valuable service.
Product development has been another major focus and resulted in development of virally safe and validated Fibrin Glue (fibrinogen concentrate & human thrombin) and FVIII, in conformance with the EP and WHO specification, and obtained Ethics Committee approval for clinical use. Today the surgeons are keen to use these products more frequently in patients because of the efficacy of the product proven in >1000 patients during last 9 years. Validation of viral inactivation was possible due to the support from Microbiology (Hospital Wing), Virology department of CMC Vellore and NARI Pune. The data on product development and validation was evaluated by a National Committee appointed by TIFAC and the product specifications received wide acceptance.
Use of fibrin matrix for Tissue Engineering and Stem Cell Research made major outcomes. Fibrin matrix was modified for each specific use; as a hemostat, drug/growth factor delivery vehicle, wound healing adjunct, and for homing and differentiation of progenitor cells into endothelial, smooth muscle, neural and epithelial cells. In each case the focus is product development and may require more effort. There are patents and publications from this area of our research as listed below.
Preparation of IgY antibodies against viper venom was also equally challenging and the data on the in vitro and in vivo effect for neutralizing the effects of venom was carried out as per the WHO regulations. The technology developed is patent-protected and ready for scale-up, clinical trial and technology transfer.
Ongoing Research Projects
a) Vascular Tissue Engineering (Funded by DBT, Govt. of India)
Project Title: Tissue Engineered Small Diameter Vascular Graft (TEVG): Fabrication & Evaluation.3 years Feb 2007 to Jan 2010.
Objective: Using a multidisciplinary approach that involve inputs from polymer chemistry, cell biology, and biomechanics, fabricate tissue engineered small diameter vascular graft and evaluate its performance experimental in animal models (Sheep carotid artery) .
Current Status: Small diameter conduits developed at dental products laboratory (DPL), for grafts are porous and degradable (e-poly caprolactone). Bioreactor is developed with two separate channels (one for EC & other for SMC) by the device testing laboratory. Tissue engineering of the graft is done at TRU using autologous circulating EPC and SMPC from each experimental animal. Well characterized graft is under evaluation in large animals. Animal experiments are done at the division of in vivo models & testing, and grafts are implanted by the vascular surgeons of cardiovascular and thoracic surgery (CVTS) of SCTIMST. Necropsy of the experimental animals and histology analysis of the explants are done at the implant biology laboratory. So it is collaborative program with multi-disciplinary research teams coordinated by Scientist-in-charge of TRU.
b) Project Title: Differentiation of Circulating Adult Stem Cells to Neurons for Regenerative Therapy of Parkinson's Disease (PD) and Spinal Cord Injury (SCI) in Animal Models. Funded by: DST(Govt. of India)
Objective: Develop an autologous cell source for neural regeneration.
Current Status:Peripheral blood mononuclear (PBMN) fraction from human blood was found to have nestin positive neural progenitor cells (NPC) in PD patients and normal controls in comparable proportion. Under very specific growth conditions, NPCs differentiated into neurons on a cell specific fibrin matrix composite. The composite was proven good as a cell delivery vehicle for Olfactory Ensheathing Cells (OEC) with improved homing in a SCI.
c) Skin Tissue Engineering: There are two ongoing projects towards construction of skin substitutes. The goal is dermal and epidermal regeneration using autologous cells in tissue engineering approach for developing patient-specific skin substitute to treat chronic diabetic and burn wounds.
Project title: Development of hemostatic scaffold for wound care (Council for Scientific and Industrial Research, Govt. of India)
Objective: Improve mechanical handling characteristic of fibrin sheet and improve it's the wound healing efficiency.
Current Status: Biodegradable polymer films with high porosity have been developed as a support membrane for fibrin sheet. Polymer film development and its evaluation for degradation property is in progress with support of dental products laboratory. Polymer-fibrin hemostat fabrication and evaluation in animal models is being planned.
Presence of keratinocyte progenitor cells (KPC) in the circulation of normal donors has been established. Differentiation of circulating progenitor cells from normal donors is standardized. Co-culture of skin derived fibroblast and keratinocyte progenitor is found efficient for propagation of KPC. Polymer-fibri hemostat seeded with autologous KPC and fibroblast is to be evaluated in diabetic model.
Project title: Bioengineered hybrid skin substitutes for burn wounds (KERALA BIOTECHNOLOGY COMMISSION, KSCSTE, Govt. of Kerala)
Objective: Development of skin substitute for treating severe burn cases.
a. Temporary wound covering materials and scaffolds for dermal tissue regeneration are being developed as part of the program at dental products laboratory.
b. In TRU: MSC isolation from adipose tissue of human source is standardized.
c. Preliminary data indicates that matrix signaling strategy may be adopted for dermal tissue generation by growing MSC on designed biomimetic matrix.
d. Curcumin Delivery
Project Title: "In vitro, and Preclinical Evaluation of Curcumin Released from Biodegradable Fibrin Matrix" funded by ICMR, Govt.of India
Objective: Development of curcumin-loaded fibrin sheet for delivery of this potential agent for control of cancer metastasis.
Current Status: Standardized process of loading curcumin on to fibrin sheet. Kinetics of release is estimated. Effect of released curcumin on cancer cell line culture is being evaluated.
Projects in which TRU faculty is a Co-investigator
a. Project Title: Quantum dots as a drug carrier for cardiovascular disease (Funded by DST, Govt. of India under Women Scientist Scheme).
Principal Investigator: Dr. Diksha Painuly
Mentors: Dr. Kalliyana Krishnan V and Dr. Lissy K. Krishnan
Co-Investigator: Dr. Anugya Bhatt
b. Project Title: Coronary Artery Desease in Young (funded by KSCSTE, Govt. of Kerala)
Principal Investigator (PI) Dr.Harikrishnan.S.
Prinicipal Co PI: Dr. Jaganmohan Tharakan
Co PI: Dr Jayakumari and Dr. Anugya Bhatt
c. Project Title: D-Dimer as a diagnostic tool for LA thrombus in rheumatic mitral stenosis.
PI-Dr.Kiron S, Cardiology, SCTIMST
Co-I, Dr. JM Tharakan, Dr. Bijulal, Dr. Harikrishnan A, Dr. Renuka Nair, and Dr. Lissy Krishnan
Basic research-Intramural support from Laboratory Fund:
Topic: Role of Integrin molecules of activated platelet membrane in differentiation of endothelial cells from PBMNCs ( Guide-Dr. Lissy Krishnan)
Current status: Activated platelet membrane immobilized on fibrin clot is found to have a signalling role on progenitors in the peripheral blood mononuclear cells (PBMNC) for converting them into endothelial lineage that might participate in the vascular regeneration, provided platelet granule contents are absent in the mileu.
Topic: Profiling of proteins released from a-granules of platelets isolated from normal volunteers and those with risk factors for atherosclerosis development (Guide-Dr. Anugya Bhatt)
Current status: Platelets isolated from human subjects with risk factors aggregate at higher rate, and release more secrtogoggin and cyclophilin as identified using Western Blotting of proteins released by activation with low concentration of thrombin.