Studies - Result of Clinical data

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Studies - Result of Clinical data Powered By Docstoc
					  The Effects of Inorganic
Polyphosphate on the Bone
Regeneration in Beagle Dogs
    Kangmoon Seo, Hongyeoul Kim1, Hoyeon Lee2,
         Changhoon Lee3, Sookyung Chae4

     Dept of Vet Med, Kangwon National Univ, 1Dept of Molecular Biology,
Institude of Oriental Med, Kyung-Hee Univ, 2Dept of Neurosurgery, Woo-Ree
Hosp, 3Dept of Neurosurgery, Korea Cancer Hosp, 4Kyung-Won Medical Inc,
                                   Korea
Introduction
What is inorganic polyphosphate?

- a linear chain of tens or many hundreds of phosphate residues
linked by high energy phosphoanhydride bonds.
- found in every cell in nature: bacterial, fungal, protozoan, plant,
and animal
- has numerous and varied biological functions depending on where
it is and when it is needed.
    . substition for ATP in kinase reactions,
    . reservoir of phosphate,
    . chelation of divalent metals, capsule of bacteria,
    . regulatory roles in growth, development, stress and deprivation.
    ? Stimulate bone regeneration(Osteoblast, osteocalcin, BMP ?)
Bone Substitute Materials
 Non-resorbable
    PMMA (Polymethyl methacrylate)
• Bioresorbable
   Autograft
   Allograft
   Hydroxyapatite
   Calcium phosphate bone cement
Ideal Bone Cement Composition

    Osteoconductive materials
               +
     Osteoinductive materials
               =
     Fast Bone Regeneration
Purpose

To evaluate the bone regeneration power of
biocompatible and biodegradable calcium
phosphate bone cement(Osteoconductive)
mixing with type 65 0.01%
polyphosphate(Osteoinductive)
Materials and Methods
• Experimental Animals
   30 Beagle dogs
   Average body wt : 12.5 kg
   Age : over 1 year
   Sex : all male

• Composition of bone cement
   β-tricalcium phosphate 41.7 wt%
   Monocalcium phosphate monohydrate 13.0 wt %
   Calcium sulfate hemihydrate 10.4 wt%
   Polyphosphate type 65 0.01%
   Distilled water 34.8%
• Bone cement cylinder
 4.8mm in diameter
 10mm in length


• Implanted site
   Distal epiphysis of femurs (5mm x 10mm)


• Experimental Groups
  Non-treated group(Control group)
  Calcium phosphate group(Ca-P group)
  Polyphosphate+calcium phosphate group (PolyP group)
A   B




C   D




E   F
• Parameter    of examination
- Radiological examination
  every weeks after operation until 6 weeks

- Histopathological examination
   5 dogs in each group at 3 weeks after operation
   5 dogs in each group at 6 weeks after operation

- Hematological & serological examination
  PCV, WBC, Hb, TP, GOT, GPT, BUN, Creatinine, Ca, P
  every weeks after operation until 6 weeks
Results

• Radiological findings
Fig 1. Sequential radiographs after drilling in distal femur of a non-treated dog.
1A: Immediate postoperation. The drilled hole is shown(arrow).
1B: 2 weeks after operation. The margin of the drilled hole(arrows) is starting to be dense.
1C: 4 weeks after operation. The distance between the arrows is the width of newly formed radiopaque area.
1D: 6 weeks after operation. Note a little changes of the density around the hole.
Fig 2. Sequential radiographs after implantation of calcium-phosphate(Ca-P) cement of the distal femur in a dog.
2A: Immediately after implantation of Ca-P cement. Arrow indicates implanted cement.
2B: 2 weeks after implantation. The distance between the arrows is the width of newly formed radiopaque area.
2C: 4 weeks after implantation. The radiopaque area become wider than before.
2D: 6 weeks after implantation. The radiopaque area appears as wide as the diameter of implanted cement(arrows)
Fig 3. Sequential radiographs after implantation of polyphosphate(PolyP) cement of the distal femur in a dog.
3A: Immediately after implantation of PolyP cement. Arrow indicates implanted cement.
3B: 2 weeks after implantation. The distance between the arrows is the width of newly formed radiopaque area.
3C: 4 weeks after implantation. The radiopaque area become wider than before.
3D: 6 weeks after implantation. The radiopaque area appears wider than 1.5 times the diameter of implanted
cement(arrows).
    Bone regeneration (mm2)   160
                              140
                              120
                              100                                       Control
                               80                                       Ca-P
                                                                        PolyP
                               60
                               40
                               20
                                0
                                    1   2       3      4       5    6
                                        W eeks after implantation



Quantitative analysis of bone regeneration in radiological findings
• Histopathological findings
Fig 4. Gross and microscopic findings 3 weeks after drilling in distal femur of a non-treated dog.
4A: Gross appearance. The drilled hole is shown(arrow).
4B: Cross-section of 4A. The bone defect cavity is still pale and empty(arrows).
4C: Histological section of 4B. The host bone trabeculae around bone defect(BD) are slightly thickened(arrows). Normal bone
marrow(BM). H&E stain, x 9.
4D: Magnification of 4C. Small trabeculaes(N) are formed in the middle of bone defect. Thickened host bone(H). H&E stain.
X40.
4E: High magnification of 4D. Many osteoclasts(white arrows) are attached to the trabeculaes and osteoblasts(blank arrows) are
occasionally seen. H&E stain. X100.
Fig 5. Gross and microscopic findings 6 weeks after drilling in distal femur of a non-treated dog.
5A: Gross appearance. The drilled hole is shown(arrow).
5B: Cross-section of 5A. The bone defect cavity is still pale(arrows).
5C: Histological section of 5B. The host bone trabeculae around bone defect(BD) are slightly thickened(arrows). H&E stain, x
9.
5D: Magnification of 5C. Many sinusoids(S) and small new bone trabeculae(N) are formed in bone defect. H&E stain. X40.
5E: High magnification of 5D. Many osteoclasts(arrows) around new bone trabeculae(N) still persist. S indicates sinusoids in
newly formed bone marrow. H&E stain. X100.
Fig 6. Histological findings 3 weeks after implantation of calcium-phosphate(Ca-P) cement
of the distal femur in a dog.
6A: Gross appearance. The implanted bone cement is shown(arrow).
6B: Cross-section of 6A. The structure around bone cement(arrows) demonstrates dense appearance.
6C: Histological section of 6B. The host bone trabeculae around implanted bone cement(BD) are thickened(arrows). H&E
stain, x 9.
6D: Magnification of 6C. New bone(N) is forming in contact with the surface of the bone cement(C). H&E stain. X40.
6E: High magnification of 6D. Many osteoclasts(white arrows) are shown in the surface of the bone cement(C) and
osteoblasts(blank arrows) are occasionally seen around new bone(N). H&E stain. X100.
Fig 7. Gross and microscopic findings 6 weeks after implantation of calcium-phosphate
(Ca-P) cement in the distal femur of a dog.
7A: Gross appearance. The implanted bone cement is shown(arrow).
7B: Cross-section of 7A. The structure around bone cement(arrows) is appeared dense.
7C: Histological section of 7B. The host bone trabeculae around implanted bone cement(BD) are thickened(arrows).
H&E stain, x 9.
7D: Magnification of 7C. The bone cement is partially resorbed(RC) and replaced with new bone on its periphery. The
host bone trabeculae(H) near bone cement are growing and thickened. H&E stain. X40.
7E: High magnification of 7D. RC indicates residual cement. H indicates host bone trabecuale. H&E stain. X100.
Fig 8. Gross and microscopic findings 3 weeks after implantation of polyphosphate(PolyP)
cement of the distal femur in a dog.
8A: Gross appearance. The implanted bone cement is shown(arrow).
8B: Cross-section of 8A. The structure around bone cement(arrows) shows dense appearance.
8C: Histological section of 8B. The host bone trabeculae around implanted bone cement(BD) are extensively
thickened(arrows). H&E stain, x 9.
8D: Magnification of 8C. New bone(N) is forming at the contacting surface of the bone cement(C). H&E stain. X40.
8E: High magnification of 8D. Many osteoblasts(blank arrows) are shown around new bone and osteoclasts(white arrows)
are occasionally seen. H&E stain. X100.
Fig 9. Gross and microscopic findings 6 weeks after implantation of polyphosphate(PolyP)
cement of the distal femur in a dog.
9A: Gross appearance. Irregularly proliferate margin(arrow) is shown around the implanted bone cement.
9B: Cross-section of 9A. Almost bone marrows around the bone cement are changed to white matrix(arrows).
9C: Histological section of 9B. The host bone trabeculae around implanted bone cement(BD) are extensively
thickened(arrows). H&E stain, x 9.
9D: Magnification of 9C. The bone cement is partially resorbed(RC) and replaced with new bone(N) at the peripheries. The
host bone trabeculae(H) near bone cement are growing and thickened. H&E stain. X40.
9E: High magnification of 9D. RC indicates residual cement. H indicates host bone trabecuale. H&E stain. X100.
• Hematological & Serological results
               80



               70                       PCV                                                                           30
                                                                                                                                             Hemoglobin
                                                                                                                      25
               60


                                                                                                                      20




                                                                                                  Hemoglobin (g/dl)
               50
PCV (%)




                                                                                                                      15
               40


                                                                                                                      10
               30
                                                                                                                                                                                         Control
                                                                                                                       5                                                                 Ca-P
                                                                             Control
               20                                                                                                                                                                        PolyP
                                                                             Ca-P
                                                                             PolyP
                                                                                                                       0
               10                                                                                                           Before       1        2          3          4          5           6
                         Before     1         2        3         4       5       6                                                                Weeks after implantation
                                            W eeks after implantation
                                                                                                                           Text-Fig 3. Hemoglobin changes after implantation of polyphosphate bone
                     Text-Fig 2. PCV changes after implantation of polyphosphate bone
                                                                                                                                       cement in a canine femur.
                                 cement in a canine femur.




                                                                                                                      14
                    30



                    25
                                            WBC                                                                       12                         TP
                                                                                                                      10

                    20
   WBC (x1000/ul)




                                                                                                                       8


                    15                                                                                 TP (g/dl)
                                                                                                                       6


                    10                                                                                                 4



                                                                                                                       2                                                               Control
                     5                                                                  Control                                                                                        Ca-P
                                                                                        Ca-P                                                                                           PolyP
                                                                                        PolyP
                                                                                                                       0
                     0
                                                                                                                             Before      1        2         3         4        5           6
                           Before       1          2        3          4       5          6
                                                                                                                                                W eeks after implantation
                                                  Weeks after implantation

                                                                                                                           Text-Fig 5. Total serum protein changes after implantation of
                         Text-Fig 4. WBC changes after implantation of polyphosphate bone                                              polyphosphate bone cement in a canine femur.
                                     cement in a canine femur.
             60                                                                                                                             140
                                                                                             Control                                                                                                                Control
                                                                                                                                                                                                                    Ca-P
             50                                  AST                                         Ca-P
                                                                                             PolyP                                          120
                                                                                                                                                                      ALT                                           PolyP



                                                                                                                                            100
             40

                                                                                                                                             80
AST (IU/L)




                                                                                                                               ALT (IU/L)
             30

                                                                                                                                             60

             20
                                                                                                                                             40


             10
                                                                                                                                             20



                  0                                                                                                                           0
                                Before       1         2          3           4        5             6                                               Before       1         2          3           4        5             6
                                                       Weeks after implantation                                                                                             Weeks after implantation

                                Text-Fig 6. AST changes after implantation of polyphosphate bone cement
                                                                                                                                                      Text-Fig 7. ALT changes after implantation of polyphosphate bone cement
                                            in a canine femur.
                                                                                                                                                                  in a canine femur.


                           70
                                                                                                                               1.4
                                                                                           Control
                           60
                                                 BUN
                                                                                           Ca-P
                                                                                           PolyP                               1.2
                                                                                                                                                                      Creatinine
                           50                                                                                                           1




                                                                                                          Creatinine (mg/dl)
             BUN (mg/dl)




                           40                                                                                                  0.8



                           30                                                                                                  0.6



                           20                                                                                                  0.4

                                                                                                                                                                                                           Control
                                                                                                                               0.2                                                                         Ca-P
                           10
                                                                                                                                                                                                           PolyP

                                                                                                                                        0
                            0
                                                                                                                                                  Before      1        2        3           4          5        6
                                  Before         1       2         3         4         5         6
                                                                                                                                                                      Weeks after implantation
                                                       W eeks after implantation
                                                                                                                                              Text-Fig 9. Creatinine changes after implantation of polyphosphate bone
                                 Text-Fig 8. BUN changes after implantation of polyphosphate bone                                                         cement in a canine femur.
                                             cement in a canine femur.
             12                                                                                        8



             11
                                    Ca                                                                 7
                                                                                                                              P
                                                                                                       6
             10




                                                                                   Phosphate (mg/dl)
                                                                                                       5
              9
Ca (mg/dl)




                                                                                                       4
              8

                                                                                                       3

              7
                                                                                                       2
                                                                        Control
              6                                                         Ca-P                                                                                        Control
                                                                        PolyP                          1                                                            Ca-P
                                                                                                                                                                    PolyP
              5
                                                                                                       0
                  Before      1        2         3        4        5        6
                                                                                                            Before      1         2        3         4        5         6
                                     W eeks after implantation
                                                                                                                                W eeks after implantation

                  Text-Fig 10. Serum calcium level changes after implantation of                           Text-Fig 11. Serum phosphate level changes after implantation of
                               polyphosphate bone cement in a canine femur.                                             polyphosphate bone cement in a canine femur.
Conclusions
Polyphosphate bone cement has a powerful
bone regeneration effects and can be used
for various bone related surgery as well as
for filling bone defects.

				
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posted:10/19/2012
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