Tissue engineering 

(TE) represent a paradigm shift in healthcare 

therapies and treatments by repairing, 

replacing, or regenerating damaged cells and 

tissues in human body. 

Chemical cues such 

as growth factors (GFs) and cytokines, and 

there successful delivery is the key 

components in TE with the ability to target 

specific tissue regeneration

Chandra, B

Cicerone, M

Giri, Jyotsnendu

Kumar, N

Tissue engineering \ud
(TE) represent a paradigm shift in healthcare \ud
therapies and treatments by repairing, \ud
replacing, or regenerating damaged cells and \ud
tissues in human body. \ud
Chemical cues such \ud
as growth factors (GFs) and cytokines, and \ud
there successful delivery is the key \ud
components in TE with the ability to target \ud
specific tissue regeneration

Giri, J

Name not available

Protein Stabilized and Sustained Deliverable Nanofiber Smart Scaffold for Multiphase Tissue Regeneration

Research Archive of Indian Institute of Technology Hyderabad

Protein stabilized and sustained deliverable nanofiber ‘smart scaffold’ for multiphase tissue regeneration Jyotsnendu Giri1*, Baishakhi Chandra2, Neha Kumar1, Marcus T Cicerone2, 1American Dental Association Foundation, 2National Institute of Standards and Technology, Gaithersburg, MD 20899, *Indian Institute of Technology, Hyderabad, IndiaStatement of Purpose: Tissue engineering (TE) represent a paradigm shift in healthcare therapies and treatments by repairing, replacing, or regenerating damaged cells and tissues in human body. Chemical cues such as growth factors (GFs) and cytokines, and there successful delivery is the key components in TE with the ability to target specific tissue regeneration. Current techniques of immobilization and loading of GF into a polymer matrix (using emulsion method) have shown only limited success in TE field due to the loss of GF’s bioactivity during fabrication process and storage, as well as generally low encapsulation efficiency, and uncontrolled release profiles.1 Thus it is difficult to provide artificial tissue microenvironment for cell growth and specific tissue differentiation. A number of approaches have been developed to ameliorate the impact of individual processing stresses, but no single approach has been available heretofore which would overcome these challenges of GF delivery from scaffold. Far from meeting this ideal, many approaches for improving one aspect of performance are neutral or deleterious to others.2        We developed novel nanoencapsulation of protein in to sugar-glass-matrix to form protein-sugar-glass-nanoparticles (SGnP) system to overcome these formidable challenges of GF (e.g., protein) stabilization and their sustained delivery (Fig. 1). SGnP could be used develop a generic platform for GF encapsulated ‘smart scaffold’ of any polymer and protein system for functional tissues regeneration by protecting protein’s activity lost against processing and storage stresses, while reaching high loads and exhibiting burst-free sustained release of protein in their active form to provide suitable tissue microenvironment for stem cell differentiation . Methods: We have encapsulated different GFs (for bone BMP 2 and cartilage BMP 7+TGFβ) into SGnP by flash-freezing and freeze-dried method reported in Ref.3. GF-SGnP (1-2 % (w/w)) is then suspended in organic solvent-based polymer (PCL/PLGA) solutions, from which tissue scaffolds were fabricated by electrospinning. Four different scaffolds are fabricated with/without encapsulating specific GF for control (no GF), bone (BMP 2), cartilage (BMP 7+ TGFβ) and bone-cartilage interfacial tissues (BMP 2 and BMP 7+ TGFβ). Protein release profile from scaffolds is also studied to check their burst free sustain release. We evaluated the performance of these smart scaffolds for specific in vitro tissue regeneration using hBMSC and cultured for 30 days using normal growth media. Gene expression, protein assay and staining are used to characterize the differentiation.  Results: We have reported that SGnP system provide excellent protection of GF from array of non-aqueous solvents with essentially no activity loss after exposure to solvents with polarity index ≤ 5, as compared to > 90% loss (and up to 99.97% loss) using emulsion methods commonly in use.3  The PCL or/and PLGA nanofiber scaffold shows sustained release profiles of GF from scaffold without significant burst release (<5 % release within day 1) as SGnP system provides excellent dispersion of encapsulated protein in the polymer matrix and consequently their diffusion based release. After 21 days of culture of stem cells (hBMSC) on these smart scaffolds in a 48 (single GF loaded scaffold) and 12 (composite scaffold) well plate using normal media, we observed formation of bone, cartilage as well as bone-cartilage interfacial tissues in a single scaffold (Figure 2).   Conclusions:  SGnP is used to develop GF encapsulated ‘smart scaffold’ system for different functional tissues regeneration. The microenvironment of GF is essential for generate specific tissues than the global concentration of GF i.e., concentration in culture media. Thus it is possible to generate suitable microenvironment of GF in a single scaffold to generate multiple interfacial tissues using the smart scaffold system where GFs are encapsulated and stabilized within SGnP system.  Reference: [1] K. Fu, A. M. Klibanov, R. Langer, Nat. Biotechnol. 18, 24. 2000.  [2]. C. Perez, at el, Journal of Pharmacy and Pharmacology 54, 301. 2002. [3] J. Giri at el, Advanced Material 23, 4861, 2011. [3] J. C. Sy, A. S. Klemm, V. P. Shastri, Advanced Materials 21, 1814, 2009. Abstract #292©2014 Society for Biomaterials

Protein stabilized and sustained deliverable nanofiber ‘smart scaffold’ for multiphase tissue regeneration 
Jyotsnendu Giri1*, Baishakhi Chandra2, Neha Kumar1, Marcus T Cicerone2, 
1American Dental Association Foundation, 2National Institute of Standards and Technology, Gaithersburg, MD 20899, 
*Indian Institute of Technology, Hyderabad, India
Statement of Purpose: Tissue engineering 
(TE) represent a paradigm shift in healthcare 
therapies and treatments by repairing, 
replacing, or regenerating damaged cells and 
tissues in human body. Chemical cues such 
as growth factors (GFs) and cytokines, and 
there successful delivery is the key 
components in TE with the ability to target 
specific tissue regeneration. Current 
techniques of immobilization and loading of 
GF into a polymer matrix (using emulsion 
method) have shown only limited success in 
TE field due to the loss of GF’s bioactivity 
during fabrication process and storage, as 
well as generally low encapsulation 
efficiency, and uncontrolled release 
profiles.1 Thus it is difficult to provide 
artificial tissue microenvironment for cell 
growth and specific tissue differentiation. A 
number of approaches have been developed 
to ameliorate the impact of individual 
processing stresses, but no single approach 
has been available heretofore which would 
overcome these challenges of GF delivery 
from scaffold. Far from meeting this ideal, 
many approaches for improving one aspect 
of performance are neutral or deleterious to others.2  
      We developed novel nanoencapsulation of protein in to 
sugar-glass-matrix to form protein-sugar-glass-nanoparticles 
(SGnP) system to overcome these formidable challenges of 
GF (e.g., protein) stabilization and their sustained delivery 
(Fig. 1). SGnP could be used develop a generic platform for 
GF encapsulated ‘smart scaffold’ of any polymer and protein 
system for functional tissues regeneration by protecting 
protein’s activity lost against processing and storage stresses, 
while reaching high loads and exhibiting burst-free sustained 
release of protein in their active form to provide suitable 
tissue microenvironment for stem cell differentiation . 
Methods: We have encapsulated different GFs (for bone 
BMP 2 and cartilage BMP 7+TGFβ) into SGnP by flash-
freezing and freeze-dried method reported in Ref.3. GF-
SGnP (1-2 % (w/w)) is then suspended in organic solvent-
based polymer (PCL/PLGA) solutions, from which tissue 
scaffolds were fabricated by electrospinning. Four different 
scaffolds are fabricated with/without encapsulating specific 
GF for control (no GF), bone (BMP 2), cartilage (BMP 7+ 
TGFβ) and bone-cartilage interfacial tissues (BMP 2 and 
BMP 7+ TGFβ). Protein release profile from scaffolds is also 
studied to check their burst free sustain release. We evaluated 
the performance of these smart scaffolds for specific in vitro 
tissue regeneration using hBMSC and cultured for 30 days 
using normal growth media. Gene expression, protein assay 
and staining are used to characterize the differentiation.  
Results: We have reported that SGnP system provide 
excellent protection of GF from array of non-aqueous 
solvents with essentially no activity loss after exposure to 
solvents with polarity index ≤ 5, as compared to > 90% 
loss (and up to 99.97% loss) using emulsion methods 
commonly in use.3  The PCL or/and PLGA nanofiber 
scaffold shows sustained release profiles of GF from 
scaffold without significant burst release (<5 % release 
within day 1) as SGnP system provides excellent 
dispersion of encapsulated protein in the polymer matrix 
and consequently their diffusion based release. After 21 
days of culture of stem cells (hBMSC) on these smart 
scaffolds in a 48 (single GF loaded scaffold) and 12 
(composite scaffold) well plate using normal media, we 
observed formation of bone, cartilage as well as bone-
cartilage interfacial tissues in a single scaffold (Figure 2).   
Conclusions:  SGnP is used to develop GF encapsulated 
‘smart scaffold’ system for different functional tissues 
regeneration. The microenvironment of GF is essential for 
generate specific tissues than the global concentration of 
GF i.e., concentration in culture media. Thus it is possible 
to generate suitable microenvironment of GF in a single 
scaffold to generate multiple interfacial tissues using the 
smart scaffold system where GFs are encapsulated and 
stabilized within SGnP system.  
Reference: [1] K. Fu, A. M. Klibanov, R. Langer, Nat. 
Biotechnol. 18, 24. 2000.  [2]. C. Perez, at el, Journal of 
Pharmacy and Pharmacology 54, 301. 2002. [3] J. Giri at 
el, Advanced Material 23, 4861, 2011. [3] J. C. Sy, A. S. 
Klemm, V. P. Shastri, Advanced Materials 21, 1814, 
2009. 
Abstract #292
©2014 Society for Biomaterials


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Protein Stabilized and Sustained Deliverable Nanofiber Smart Scaffold for Multiphase Tissue Regeneration

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