bone regeneration

Approaches For
Bone Regeneration
Presented By:
Mr. Mahesh A. Marathe.
M. Pharm. Sem I (P’ceutics.)
Roll No. 516
A
Seminar on
Guide:
Dr. K. N Gujar.
Mrs. V. M. Gambhire.
SINHGAD COLLEGE OF PHARMACY ,VADGAON , PUNE

MaheshA.Marathe.16/11/2016
2
Bone :
 They are made up of living cells .
Minerals, vitamins, proteins are helpful for growth and repair
bone.
Bone tissue compose of :
Harder outer layer (Cortical/compact bone)
Inner spongy layer (trabecular/cancellous
bone).
https://www.iofbonehealth.org/introduction-bone-biology-all-about-our-bones,access on 3/11/2016

3
Bone cells are composed of :
Osteoblasts and osteocytes
osteoclasts
Osteiods
Inorganic mineral salts
Type of Bones :
Woven bone
Lamellar bone
https://www.iofbonehealth.org/introduction-bone-biology-all-about-our-bones,access on 3/11/2016

4
Bone regeneration is process which response to any
injury like accidental fracture, development of continuous
remodelling of bone, disease condition like osteoporosis
and after surgery like arthoplasty, etc
Bone regeneration process involves certain processes like
hematoma formation, callus formation, callus ossification and
bone remodelling
Madhukar R. W., Dilip R . (2013)Bone Regeneration and Repair: Current and Future Aspects,
International Journal of Science and Research (IJSR) ,volume 4 (10),pp.351-355

7
There are over 15 million fracture cases globally and an
estimated 2.2 million bone graft procedures are performed
annually to promote fracture healing or to fill defects .
Sometimes the common solutions, such as Autografts &
Allografts are not suitable due to the donor site morbidity,
disease transmissions, and, for the implants case, failure may
occur after transplantation of tissue.
Carolina G., Frederico R.,Paulo R., (2015),A biomechanical approch for bone regeneration
inside scaffolds,procedia engineering,volume 4(2016),pp82-89

8
Complication
in Bone
Regeneration
Fracture
shortening
Fracture non
unions
Valgus
deformity

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Due to these complication or deformities at particular site of
facture we need mechanical support for that particular area.
Mechanical support /stability of fracture we need some
structural backbone for support like,
a) cast
b) Rod
c) External fixator
d) Plates & screw

11
scaffolds
polymeric
Natural
Bone morphogenic proteins
(BMPs)
Mesenchymal cells
&Growth factors

12
Natural- or biologic-based materials are taken from
biologic-based tissues.
 scaffolding act as Extra Cellular Matrix for cells.

13
Biodegradability
Regenerative
Biocompatible

14
 Delivery of cells to desired sites,
 scaffold should enhance regenerative capability,
 Must provide correct geometry to define and
maintain space for tissue regeneration.
Rozalia d.,Elna j.,dennis m.,(2011),bone regeneration:current concepts and future
directions,journal of cellular and molecular medicine,volume 15(2011),pp719-738

15
POLYMERIC :
Material :chitosan, alginate, etc.
Delivery system : hydrogels, fibres, thin films
Natural :
Material :collagen, elastin, fibrin, etc.
Delivery system : Hydrogels
Ceramic :
Material :calcium phosphate cement
Delivery system :Porous matrix structures

16
Bone morphogenetic proteins (BMPs) are a group of
growth factors also known as cytokines .
 Originally discovered by their ability to induce the
formation of bone and cartilage.
. Recombinant human BMPs are used in orthopedic
applications such as spinal fusions, nonunions, etc

17
BMPs
Induce
differentiations
towards
osteoblast
Potent
osteoinductive
factor
Induce
mitogenesis of
MSCs
Induce
osteoprogenitors

18
Mesenchymal stem cells (MSCs) : Multipotent
stromal cells that made up of different cell like:
osteoblasts (bone cells), chondrocytes (cartilage cells)
and adipocytes (fat cells).
Stromal cells are connective tissue cells that form the
supportive structure to bone .

19
Viktor Tollemar a,b,c, Zach J. Collier a,b, Maryam K. ,et.al, Stem cells,(2013), growth factors and scaffolds
in craniofacial regenerative medicine .volume 3, pp59

20Viktor Tollemar a,b,c, Zach J. Collier a,b, Maryam K. ,et.al, Stem cells,(2013), growth factors and scaffolds
in craniofacial regenerative medicine .volume 3, pp59

Yurong C , Juhong Y, Subhas C. Kundu b, Juming Y (2016), Multifunctional nano-hydroxyapatite and alginate/gelatin
based sticky gel composites for potential bone regeneration, Materials Chemistry and Physics, pp 1-7
21
Objective :To improve the fixations of the implant and implant-bone
integration after joint arthroplasty from locally preventing inflammation
and promoting the bone regeneration.

J.H. Yu, X.B. Chu, Y.R. Cai, P.J. Tong, J.M. Yao, Preparation and characterization of antimicrobial nano-hydroxyapatite
composites, Mater. Sci. Eng. C 37 (1) (2014) 54e59.
22
Synthesis of drug loaded nanohydroxyapatite (nHAP)
50ml CaCl2 solution in 150ml of silk sericin solution
Add 50ml Na2HPO4 dropwise in above solution
pH should be made 10 by using 1M NaOH solution
Precipitates collected after 2 hours by centrifugation
Lyophilized to obtain nHAP particles
50oC with agitation30 min
Rinse with ethyl alcohol (3 times)

J.H. Yu, X.B. Chu, Y.R. Cai, P.J. Tong, J.M. Yao, Preparation and characterization of antimicrobial nano-
hydroxyapatite composites, Mater. Sci. Eng. C 37 (1) (2014) 54e59.
23
Preparation of VAN-loaded nHAP :
100mg nHAP in 2 ml SBF (pH – 7.4)containing 5 mg
vancomycin chloride (VAN)
At vacuum, at ambient temperature for
24 hours

24
PREPARATION COMPOSITE
GEL :
No
drug-loaded
nHAP/OSA/G
T gel
VAN-loaded
nHAP/OSA/G
T gel
rhBMP-2
loaded
nHAP/OSA/G
T gel
Both VAN and
rhBMP-2
loaded
nHAP/OSA/G
T gel
HOG VHOG BHOG VBHOG

25
Four gel column (1ml each)
Soak in 5ml SBF(pH7.4) at 370 C resp.
After 1,3,5 &7 days supernantant was
collected & mixed with S. aureus
bacteria cultured media.
After 24 hr. ,culture forming unit (CFU)
evaluated for sustained bacterial effect.

based sticky gel composites for potential bone regeneration, Materials Chemistry and Physics, pp 1-7 26
MG63 cell bone related cell model
50 microgram composite gel injected into 96-well plates
Use blank as a control
Gel were wash by PBS(pH7.4)
Cells on gel were evaluated by MTT test.

27

29
Rat anesthesized with 4% chloral hydrate
On middle part of skull bone defect of 0.2 mm size
generated using table electric dental engine
Column sample of HOG & VBHOG was inserted.

30
Each skull was
fixed in 4% PBS
(pH7.4) buffer
formalin
Keep skull for
over period of
night
decalcification
Embedded with
paraffin
To obtain thin
size slices
Obtain slices of 3
micrometer
Dyed with
heamatoxylin
eosin
Observe under
optical
microscopy

31
XRD & TEM

32
Adsorption rate & cumulative release
Mahesh A. Marathe.
16/11/2016

Morphology and stickiness

34
Colony count for
determination of
antibacterial
activity.
MG-63 cells
(bone related
cells) grown to
study the cell
proliferation
using MTT test

36

37
 A multifunctional composite with rhBMP-2, antibiotic
loaded nano-hydroxyapatite and alginate/gelatin sticky gel
formulated . It results in reduce the mobilization of the
composite on the fractured bone surface structure.
Sticky gel composite shows sustained release action.
The composite shows antimicrobial effect and ability to
promote the bone cell proliferation.
The composite reduce the inflammation, promotes the
formation of new bones and blood vessels in vivo.

 Yu J.H., , Y.R. CaCHU X.B., TONG P.J., YAO J.M., Preparation and
characterization of antimicrobial nano-hydroxyapatite composites, Mater.
Sci. Eng. C 37 (1) (2014) 54-59.
39
Yurong C , Juhong Y, Subhas C. Kundu b, Juming Y (2016),
Multifunctional nano-hydroxyapatite and alginate/gelatin based sticky gel
composites for potential bone regeneration, Materials Chemistry and
Physics, pp 1-7..Downloadspharmaceutics seminarseminar topic.pdf
 Madhukar R. W., Dilip R . (2013)Bone Regeneration and Repair:
Current and Future Aspects, International Journal of Science and
Research (IJSR), 4 (10),pp.351-355

R15(2011),pp719-738
40
Viktor Tollemar a,b,c, Zach J. Collier a,b, Maryam K.et.al,(2013) Stem
cells, growth factors and scaffolds in craniofacial regenerative medicine
.volume 3, pp.59..Downloads1-s2.0-S2352304215000653-main.pdf
Xu Y.T, Yu X.X, Gu Z.P., X. ,(2012) study of a novel crosslinking
reagent (alginate dialdehyde) for biological tissue fixation, Polym.,pp.
1589-1595.
Carolina G., Frederico R.,Paulo R., (2015),A biomechanical approch for
bone regeneration inside scaffolds, procedia engineering,volume
4(2016),pp82-89

bone regeneration

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