Division of Thrombosis Research

FacultyServices Projects PublicationsResearch Staff

This Division carries out the evaluation of material - blood interaction. The tests used for evaluation of blood compatibility are classified into five categories according to the process or system being tested such as Thrombosis, Coagulation, Platelets and platelet function, hematology and immunology. In addition, quantitative estimation of protein adsorption and endothelial cell adhesion and proliferation on surfaces with ability to resist shear stress under, which are important indicators of blood compatibility are also being carried out. Other activities include the development of new reagents and tests for evaluation of blood - material interaction, development of haemostatic bioadhesives for clinical applications, studies on modification of blood contacting biomaterials to reduce thrombogenicity by tissue engineering, in combination with bioadhesives.


The division has three major functions.

Testing Service
  1. In vitro blood compatibility of biomaterials
  2. In vitro biomaterial/drug response to vascular cells
  3. Consultancy (free) provided for Industrial and Academic customers
Product (Bio therapeutics) Development
  1. Fibrin Glue (fibrinogen concentrate & thrombin), FVIII, as per EP specifications
  2. Fibrin Sheet as hemostat/wound healing, biodegradable matrix
  3. Antibodies against viper venom as per WHO requirements
  1. Use bio mimetic extra cellular matrix composition for differentiation of circulating adult stem cells for regenerative therapy (Endothelial cell, Smooth muscle cells, Neurons & Keratinocytes).
  2. Development of mesenchymal stem cells from adipose tissue & bone marrow for dermal regeneration
  3. Tissue engineered small diameter vascular graft
  4. Skin tissue engineering.
  5. Clinical research on thrombosis & thrombolysis
  6. Role of circulating monocytes in atherosclerosis
  7. Platelet function & its role in cardiovascular diseases
  8. Proteomic profiling of platelets

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.

Test service
  1. Laboratory for Interaction of Materials with Blood (LIMB) was established as a part of Thrombosis Research Unit.
  2. Management of test services for Interaction of Materials with blood (ISO10993: Part 4), was done effectively with implementation of quality system as per ISO17025.
  3. Initiated participation in a Proficiency testing program accredited by NATA (Australia) and continued for past 8 years with consistent and acceptable results for the hematology and coagulation tests that are standardized in TRU.
  4. A floating team is continuously trained who contributed for test services for the past 8 years.
  5. Fourteen tests listed in ISO 10993 & five biochemical parmeters relevant for blood bag testing are validated and Quality System (ISO17025) was implemented in the laboratory for Interaction of Materials with Blood and up graded the blood compatibility test facility for obtaining accreditation by COFRAC (France)
  6. Nearly 30 tests are standardized and offered to researchers within the Institute and other research organization and major industries. Nature of tests offered include biochemical, hematological and cell culture analysis for blood-/cyto- compatibility evaluations using human blood, human endothelial cells and smooth muscle cells.
  7. Timely consultancy is provided to investigators from within the Institute/external research organizations/Industries for selection of appropriate tests for blood compatibility evaluation and for interpretation of the data.
  8. Extends platelet function testing by accepting samples routinely from patients of Medical college hospitals, and other private hospitals as this test for diagnosis of hemorrhagic disorders is not available anywhere in the state of Kerala.

Please contact or for information.

Product Developments
  1. Development of Fibrin Glue (current status)
  2. Small pool plasma fractionation is on-going with continuous supply of the two-component product to Neurosurgery and Vascular surgery theatres of SCTIMST for clinical use.
    Scale-up facility is getting ready for processing of large (100 bag) pools for multicentric use and commercialization.
    Products planned for isolation are (i) fibrinogen concentrate for fibrin glue; (ii) thrombin for fibrin glue; (iii) Factor VIII concentrates
  3. Fibrin matrix as a Hemostat & drug carrier (Current status)
  4. Established the use of lyophilized fibrin sheet as a hemostat with wound healing properties using liver stab injury skin flap injury models and rabbit ear injury model.
    Sustained release of antibiotics, growth factors and anti inflammatory molecules such as curcumin from fibrin sheet has been proven in vitro.
  5. Development of Anti venom (current status)
  6. Acute toxicity of IgY in guinea pig and Balb/c mice.
    Dose calculation of the IgY (ED50) required for reversing the effects of venom has been calculated using animal models.
    Efficacy of the product has been proven in envenomed rabbit model.
    Safety of the product for intravenous administration of antivenom has been proven in rabbit models.
  7. New Born Calf Serum for Tissue Culture
  8. Process has been standardized for collection of serum from new born calf.
    Dialyzed serum is dispensed and kept under -80oC and added in tissue culture medium
    Proliferation potential of the serum is proven
    Sterility is proven
    Identified an entrepreneur for scale-up of the process and commercialization
Ongoing Projects

Scale-up and Multi centric trial of Fibrin Glue & FVIII (Financial support from the Institute-TDC)

Technology Proving Project:

The primary goal is to prove that a small-scale plasma fractionation facility can be set up for producing fibrin glue and FVIII. The facility is planned in such a way that it can be replicated by other Blood Banks. Total area projected is ~800 sq.ft, out of which ~130 sq. ft. is Class 100, ~150 sq.ft is Class 1000 and the rest is Class 100,000.


i. Prepare 100 units of (1ml fibrinogen concentrat+1 ml human thrombin) fibrin glue kits per month by processing 50 plasma bags.

ii. Prepare 50 vials (150IU/vial) of lyophilized FVIII per month by processing 50 plasma bags per month by processing 50 plasma bags.

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.

Current Status:

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.