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A REVIEW : NANOFIBERS APPLICATION




                     ANIL KUMAR
          M.PHIL./PH.D. NANOSCIENCE 2012-13
 CENTRE FOR NANOSCIENCE, CUG, GANDHINAGAR SEC-30,
                       GUJARAT
                kmr.nano@indiatimes.com
Nanotechnology
• The study of control of matter on an atomic and molecular
  scale.
   • Deals with structures the size of 100 nanometers or
     smaller (1 nm = 1/1,000,000,000 m or 10-9 m).
   • Involves engineering on a small scale to create smaller,
     cheaper, lighter, and faster devices that can do more
     things with less raw materials.
NANOFIBERS - INTRODUCTION



ECM fibers ~ 50-500 nm in diameter
Cell ~ several-10 um
Fibers 1-2 orders of magnitude < cell
single cell contacts thousands of fibers



three techniques to achieve Nano-fibers scale

- Self Assembly
-Phase Separation
-Electro-spinning
Techniques To Achieve Nano-fibers For TE

Self-assembly




                Phase separation




                                   Electrospinning
Collagen Fibers
Formed Of Parallel Fibrils
High Modulus Of Elasticity




                                          1.5 nanometers in diameter




      300 nanometers long;

             20 collagen types that exist in animal tissue
Assembly Of Collagen Fibers
Elastin Fibers
• An amorphous protein
• Much lower modulus
  of elasticity than
  collagen
• Primary constituent of
  many ligaments
• Crosslinked
  tropoelastin
NANOFIBERS: SELF-ASSEMBLY



Definition: spontaneous organization into stable structure without
covalent bonds

Biologically relevant processes
- Cellulose,Lipids, DNA, RNA, protein organization
- can achieve small diameter


Example: peptide-amphipathics
- hydrophobic tail
- cysteine residues  disulfide bonds
Self-assembly

•   Relies on non-covalent interactions to achieve
    spontaneously assembled 3D structure.
•   Biopolymers such as peptides and nucleic acids
    are used. Example is peptide-amphiphile (PA)
•   (A) Chemical structure of (PA)
•   (B) Molecular model of the PA showing the
    narrow hydrophobic tail to the bulkier peptide
    region
•   (C) Schematic of PA molecules into a cylindrical   peptide-amphiphile
    micelle.5                                                               Nanofiber
Phase Separation
    Definition: thermodynamic separation of polymer solution into polymer-rich and
    polymer-poor layers
•   This process involves dissolving of a polymer in a
    solvent at a high temperature followed by a liquid–
    liquid or solid–liquid phase separation induced by
    lowering the solution temperature
•   Capable of wide range of geometry and dimensions
    include pits, islands, fibers, and irregular pore
    structures
•   Simpler than self-assembly




                                                          a) powder, b) scaffolds with continuous network, c) foam with closed pores.4
Elastin Is An “Entropic Spring”
• ΔG = ΔH – TΔS
• ΔH = enthalpy changes, which don’t normally happen in solvents.
• ΔS = entropy changes…
  changes in the degree of
  order
• Stretching a polymer
  increases it’s order, and
  makes ΔS negative.
• ΔG is then positive, and
  unfavorable.
NANOFIBERS: ELECTROSPINNING

Definition: electric field used to draw polymer stream out of
solution



                                       A- polymer solution in syringe
                                       B- metal needle
                                       C- high voltage applied to need
                                       D- electric field overcomes
                                       solution surface tension;
                                       polymer stream generated
                                        E- fibers 1) collected and
                                        2) patterned on plate
NANOFIBERS: ELECTROSPINNING

- multiple polymers can be combined at
         1) monomer level
         2) fiber level
         3) scaffold level
- control over fiber diameter
         alter concentration/viscosity
- fiber length unlimited
- control over scaffold architecture
         target plate geometry
         target plate rotational speed
Current approaches combined techniques
- usually electrospinning + phase separation
- fibers woven over pores
NANOFIBERS: OVERVIEW
ELECTROSPINNING POLYMERS

Chemical Synthetics
- Polyglycolic acid (PGA)
- Polylactic acid (PLA)
- PGA-PLA
- Polydioxanone (PDO)
- Polycaprolactone
- PGA-polycaprolactone
- PLA-polycaprolactone
- Polydioxanone-polycaprolactone
Natural
- Elastin
- Gelatin collagen
- Fibrillar collagen
- Collagen blends
- Fibrinogen
- Heamoglobine
POLYGLYCOLIC ACID (PGA)

Properties Parameters
- surface area to volume ratio
- biocompatible
- consistent mechanical properties
         hydrophilic
         predictable bioabsorption
- electrospinning yields diameters ~ 200 nm
Model of Surface-to-volume
                     Comparisons…
    Single Box Ratio
          6 m2
               = 6 m2/m3
          1 m3
   Smaller Boxes Ratio
      12 m2
             = 12 m2/m3
      1m  3




• Neglecting spaces between the smaller boxes, the volumes of the box on
  the left and the boxes on the right are the same but the surface area of the
  smaller boxes added together is much greater than the single box.
POLYGLYCOLIC ACID (PGA)




Random fiber collection (L), aligned collection (R)
POLYLACTIC ACID (PLA) – 200 nm

- aliphatic polyester
- L optical isomer used
         by-product of L isomer degradation = lactic acid
- methyl group decreases hydrophilicity
- half-life ideal for drug delivery



Parameters (similar to PGA)
- surface area to volume ratio
- spinning orientation affects scaffold elastic modulus


Compare to PGA
- low degradation rate
- less pH change
POLYLACTIC ACID (PLA) – 200 nm

Thickness controlled by electrospin solvent




Chloroform solvent (L) ~ 10 um, HFP (alcohol) solvent (R) ~ 780 nm

                  Both fibers randomly collected
PGA+PLA = PLGA

- tested composition at 25-75, 50-50, 75-25 ratios
- degradation rate proportional to composition
- hydrophilicity proportional to composition




Recent Study
- delivered PLGA scaffold cardiac tissue in mice
- individual cardiomyocytes at seeding
- full tissue (no scaffold) 35 weeks later
- scaffold loaded with antibiotics for wound healing
POLYDIOXANONE (PDO)


- crystalline (55%)
- degradation rate between PGA/PLA ,close to 40-60 ratio
- shape memory
- modulus – 46 MPa; compare: collagen – 100 Mpa, elastin – 4 MPa




Advantages
- PDO ½ way between collagen/elastin, vascular ECM components
- cardiac tissue replacement (like PLGA)
- thin fibers (180nm)
POLYCAPROLACTONE (PCL)

- highly elastic
- slow degradation rate (1-2 yrs)
- > 1 um
- similar stress capacity to PDO, higher elasticity


 Applications Loaded with:
- collagen  cardiac tissue replacement
- calcium carbonate  bone tissue strengthening
- growth factors  mesenchymal stem cell differentation
POLYCAPROLACTONE + PLA



Clinical Applications
- several planned
- all vasculature tissue
- high PLA tensile strength react (constrict) to sudden pressure
    increase
- increased elasticity with PCL passively accommodate large fluid
    flow
ELASTIN


- highly elastic biosolid (benchmark for PDO)
- hydrophobic
- present in: vascular walls, skin

Synthesis of Biosolid?
- 81 kDa recombinant protein (normal ~ 64 kDa)
- repeated regions were involved in binding
- 300 nm (not as small as PDO ~ 180 nm)
- formed ribbons, not fibers – diameter varies
COLLAGENS: GELATIN

- highly soluble, biodegradable (very rapid)
- current emphasis on increasing lifespan


                    COLLAGENS: FIBRIL FORMING

Type I
- 100 nm (not consistent)
- almost identical to native collagen (TEM)
- present is most tissues


Type II
- 100-120 nm (consistent)
- found in cartilage
- pore size and fiber diameter easily controlled by dilution
COLLAGENS BLENDS

In context: vasculature
- intima – collagen + elastin
- media – thickest+ elastin+collagen
- adventia – collagen
RECONSTRUCTING THE MEDIA


            - SMC seeded into tube
            - average fiber ~ 450 nm
                    slightly larger ECM fibers
            - incorporation of GAG
                    carbohydrate ECM
                    collagen crosslinker
                    mediate signalling




            - cross section of tube wall
            - 5 day culture
                     complete scaffold
            infiltration
FIBRINOGEN

- smallest diameter (both synthetic and bio)
         80, 310, 700 nm fibers possible
- high surface area to volume ratio
         increase surface interaction
         used in clot formation

                                           Stress capacity comparable
                                           to collagen (80%)
HEMOGLOBIN

- hemoglobin mats
- clinical applications:
         drug delivery
         hemostatic bandages
- fiber sizes 2-3 um
- spun with fibrinogen for clotting/healing
- high porosity = high oxygenation
Application of Nano-fibers


•   NanoFibers   in Tissue Engineering
•   NanoFibers   in Industrial composite
•   NanoFibers   in Medicals
•   NanoFibers   in Filtration
Cell and Tissue Engineering, Nanotechnology


         Cells                 Scaffolds


                   Tissue
                 Engineering



      Bioreactors                Signals
Tissue Engineering (TE)
• Scaffolds
    Biomaterials, which may be natural or
    artificially derived, providing a platform for
    cell function, adhesion and transplantation

• Cells
    Any class of cell, such as stem or
    mesenchymal cell

• Signals
     Proteins and growth factors driving the
     cellular functions of interest

• Bioreactor
     System that supports a biologically active
     environment (ex. Cell culture)



                                                     Image sourse: Stke.sciencemag.org, Nature.com
Cosmetic Application (Nanocellulose
Pack)
Fibers use in Filter
          Keratin from Wool                           Nanofibre Non-Wovens


  Properties of regenerated wool                      Nanofibre non-wovens properties
              keratin
                                                            High surface/volume ratio
 Heavy metals absorption [1]                                    High porosity
 Formaldeyde absorption [2]



                                   Filtration System
                                      Air Cleaning
                          Water depuration: especially removal of
                           ultrafine particles and heavy metals
                                        adsorption
Conclusion

COOH           COOH
                                     Cells
       NH2
                                                            Functional Tissue
                      NH2
                            COOH     Adhesive Proteins
             NH2
                               NH2
                                     Functionalized Nanofiber
                                     Nanofibrous Scaffold

• Physical, Chemical and Biological mimicking enable
  various tissue engineering application.
• Tissue engineering holds the promise to develop powerful
  new therapies "biological substitutes" for structural and
  functional disorders of human health that have proven
  difficult or impossible to address successfully with the
  existing tools of medicine.
Thank You so much

                Dean’s :-
  Prof. M.H. Fullekar & Prof. Mansingh

            Chair Persons:-
               Dr. P. Jha
             Dr. B. Pathak
             Dr. D. Mandal

               Friend's :-
        M.Phil./Ph.D. Students

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Anil ppt review nanofibers application

  • 1. A REVIEW : NANOFIBERS APPLICATION ANIL KUMAR M.PHIL./PH.D. NANOSCIENCE 2012-13 CENTRE FOR NANOSCIENCE, CUG, GANDHINAGAR SEC-30, GUJARAT kmr.nano@indiatimes.com
  • 2. Nanotechnology • The study of control of matter on an atomic and molecular scale. • Deals with structures the size of 100 nanometers or smaller (1 nm = 1/1,000,000,000 m or 10-9 m). • Involves engineering on a small scale to create smaller, cheaper, lighter, and faster devices that can do more things with less raw materials.
  • 3. NANOFIBERS - INTRODUCTION ECM fibers ~ 50-500 nm in diameter Cell ~ several-10 um Fibers 1-2 orders of magnitude < cell single cell contacts thousands of fibers three techniques to achieve Nano-fibers scale - Self Assembly -Phase Separation -Electro-spinning
  • 4. Techniques To Achieve Nano-fibers For TE Self-assembly Phase separation Electrospinning
  • 5. Collagen Fibers Formed Of Parallel Fibrils High Modulus Of Elasticity 1.5 nanometers in diameter 300 nanometers long; 20 collagen types that exist in animal tissue
  • 7. Elastin Fibers • An amorphous protein • Much lower modulus of elasticity than collagen • Primary constituent of many ligaments • Crosslinked tropoelastin
  • 8. NANOFIBERS: SELF-ASSEMBLY Definition: spontaneous organization into stable structure without covalent bonds Biologically relevant processes - Cellulose,Lipids, DNA, RNA, protein organization - can achieve small diameter Example: peptide-amphipathics - hydrophobic tail - cysteine residues  disulfide bonds
  • 9. Self-assembly • Relies on non-covalent interactions to achieve spontaneously assembled 3D structure. • Biopolymers such as peptides and nucleic acids are used. Example is peptide-amphiphile (PA) • (A) Chemical structure of (PA) • (B) Molecular model of the PA showing the narrow hydrophobic tail to the bulkier peptide region • (C) Schematic of PA molecules into a cylindrical peptide-amphiphile micelle.5 Nanofiber
  • 10. Phase Separation Definition: thermodynamic separation of polymer solution into polymer-rich and polymer-poor layers • This process involves dissolving of a polymer in a solvent at a high temperature followed by a liquid– liquid or solid–liquid phase separation induced by lowering the solution temperature • Capable of wide range of geometry and dimensions include pits, islands, fibers, and irregular pore structures • Simpler than self-assembly a) powder, b) scaffolds with continuous network, c) foam with closed pores.4
  • 11. Elastin Is An “Entropic Spring” • ΔG = ΔH – TΔS • ΔH = enthalpy changes, which don’t normally happen in solvents. • ΔS = entropy changes… changes in the degree of order • Stretching a polymer increases it’s order, and makes ΔS negative. • ΔG is then positive, and unfavorable.
  • 12. NANOFIBERS: ELECTROSPINNING Definition: electric field used to draw polymer stream out of solution A- polymer solution in syringe B- metal needle C- high voltage applied to need D- electric field overcomes solution surface tension; polymer stream generated E- fibers 1) collected and 2) patterned on plate
  • 13. NANOFIBERS: ELECTROSPINNING - multiple polymers can be combined at 1) monomer level 2) fiber level 3) scaffold level - control over fiber diameter alter concentration/viscosity - fiber length unlimited - control over scaffold architecture target plate geometry target plate rotational speed Current approaches combined techniques - usually electrospinning + phase separation - fibers woven over pores
  • 15. ELECTROSPINNING POLYMERS Chemical Synthetics - Polyglycolic acid (PGA) - Polylactic acid (PLA) - PGA-PLA - Polydioxanone (PDO) - Polycaprolactone - PGA-polycaprolactone - PLA-polycaprolactone - Polydioxanone-polycaprolactone Natural - Elastin - Gelatin collagen - Fibrillar collagen - Collagen blends - Fibrinogen - Heamoglobine
  • 16. POLYGLYCOLIC ACID (PGA) Properties Parameters - surface area to volume ratio - biocompatible - consistent mechanical properties hydrophilic predictable bioabsorption - electrospinning yields diameters ~ 200 nm
  • 17. Model of Surface-to-volume Comparisons… Single Box Ratio 6 m2 = 6 m2/m3 1 m3 Smaller Boxes Ratio 12 m2 = 12 m2/m3 1m 3 • Neglecting spaces between the smaller boxes, the volumes of the box on the left and the boxes on the right are the same but the surface area of the smaller boxes added together is much greater than the single box.
  • 18. POLYGLYCOLIC ACID (PGA) Random fiber collection (L), aligned collection (R)
  • 19. POLYLACTIC ACID (PLA) – 200 nm - aliphatic polyester - L optical isomer used by-product of L isomer degradation = lactic acid - methyl group decreases hydrophilicity - half-life ideal for drug delivery Parameters (similar to PGA) - surface area to volume ratio - spinning orientation affects scaffold elastic modulus Compare to PGA - low degradation rate - less pH change
  • 20. POLYLACTIC ACID (PLA) – 200 nm Thickness controlled by electrospin solvent Chloroform solvent (L) ~ 10 um, HFP (alcohol) solvent (R) ~ 780 nm Both fibers randomly collected
  • 21. PGA+PLA = PLGA - tested composition at 25-75, 50-50, 75-25 ratios - degradation rate proportional to composition - hydrophilicity proportional to composition Recent Study - delivered PLGA scaffold cardiac tissue in mice - individual cardiomyocytes at seeding - full tissue (no scaffold) 35 weeks later - scaffold loaded with antibiotics for wound healing
  • 22. POLYDIOXANONE (PDO) - crystalline (55%) - degradation rate between PGA/PLA ,close to 40-60 ratio - shape memory - modulus – 46 MPa; compare: collagen – 100 Mpa, elastin – 4 MPa Advantages - PDO ½ way between collagen/elastin, vascular ECM components - cardiac tissue replacement (like PLGA) - thin fibers (180nm)
  • 23. POLYCAPROLACTONE (PCL) - highly elastic - slow degradation rate (1-2 yrs) - > 1 um - similar stress capacity to PDO, higher elasticity Applications Loaded with: - collagen  cardiac tissue replacement - calcium carbonate  bone tissue strengthening - growth factors  mesenchymal stem cell differentation
  • 24. POLYCAPROLACTONE + PLA Clinical Applications - several planned - all vasculature tissue - high PLA tensile strength react (constrict) to sudden pressure increase - increased elasticity with PCL passively accommodate large fluid flow
  • 25. ELASTIN - highly elastic biosolid (benchmark for PDO) - hydrophobic - present in: vascular walls, skin Synthesis of Biosolid? - 81 kDa recombinant protein (normal ~ 64 kDa) - repeated regions were involved in binding - 300 nm (not as small as PDO ~ 180 nm) - formed ribbons, not fibers – diameter varies
  • 26. COLLAGENS: GELATIN - highly soluble, biodegradable (very rapid) - current emphasis on increasing lifespan COLLAGENS: FIBRIL FORMING Type I - 100 nm (not consistent) - almost identical to native collagen (TEM) - present is most tissues Type II - 100-120 nm (consistent) - found in cartilage - pore size and fiber diameter easily controlled by dilution
  • 27. COLLAGENS BLENDS In context: vasculature - intima – collagen + elastin - media – thickest+ elastin+collagen - adventia – collagen
  • 28. RECONSTRUCTING THE MEDIA - SMC seeded into tube - average fiber ~ 450 nm slightly larger ECM fibers - incorporation of GAG carbohydrate ECM collagen crosslinker mediate signalling - cross section of tube wall - 5 day culture complete scaffold infiltration
  • 29. FIBRINOGEN - smallest diameter (both synthetic and bio) 80, 310, 700 nm fibers possible - high surface area to volume ratio increase surface interaction used in clot formation Stress capacity comparable to collagen (80%)
  • 30. HEMOGLOBIN - hemoglobin mats - clinical applications: drug delivery hemostatic bandages - fiber sizes 2-3 um - spun with fibrinogen for clotting/healing - high porosity = high oxygenation
  • 31. Application of Nano-fibers • NanoFibers in Tissue Engineering • NanoFibers in Industrial composite • NanoFibers in Medicals • NanoFibers in Filtration
  • 32. Cell and Tissue Engineering, Nanotechnology Cells Scaffolds Tissue Engineering Bioreactors Signals
  • 33. Tissue Engineering (TE) • Scaffolds Biomaterials, which may be natural or artificially derived, providing a platform for cell function, adhesion and transplantation • Cells Any class of cell, such as stem or mesenchymal cell • Signals Proteins and growth factors driving the cellular functions of interest • Bioreactor System that supports a biologically active environment (ex. Cell culture) Image sourse: Stke.sciencemag.org, Nature.com
  • 35. Fibers use in Filter Keratin from Wool Nanofibre Non-Wovens Properties of regenerated wool Nanofibre non-wovens properties keratin  High surface/volume ratio  Heavy metals absorption [1]  High porosity  Formaldeyde absorption [2] Filtration System Air Cleaning Water depuration: especially removal of ultrafine particles and heavy metals adsorption
  • 36. Conclusion COOH COOH Cells NH2 Functional Tissue NH2 COOH Adhesive Proteins NH2 NH2 Functionalized Nanofiber Nanofibrous Scaffold • Physical, Chemical and Biological mimicking enable various tissue engineering application. • Tissue engineering holds the promise to develop powerful new therapies "biological substitutes" for structural and functional disorders of human health that have proven difficult or impossible to address successfully with the existing tools of medicine.
  • 37. Thank You so much Dean’s :- Prof. M.H. Fullekar & Prof. Mansingh Chair Persons:- Dr. P. Jha Dr. B. Pathak Dr. D. Mandal Friend's :- M.Phil./Ph.D. Students

Editor's Notes

  1. ABC blocks help students to understand the concept of surface to volume ratios. They need to see the additional surfaces!
  2. Me-zen-ki-mel