Microbiologic  effects  of  irrigation  using  functional  waters  (the  strongly  acidic  electrolyzed  water  and  the  strongly  alkaline  electrolyzed  water)  in  peri-­implant  sulcus
Kazuhiko  Inoue1,2,3),Shiota  Makoto2),  Masanori  Nashimoto1),  Syoji  Enomoto2),  Syohei  Kasugai2)
General  Implant  Reseach  Center1)     Oral  Implantology  &  Regenerative  Dental  Medicine，Tokyo  Medical  and  Dental  University2)
Department  of  Translational  Research，Tsurumi  University  School  of  Dental  Medicine3)

Ⅰ  Purpose：Peri-­implant  mucosa  does  not  have  connective  tissue  adhesion  with  implants  and  its  epithelial  attachment  is  weaker  than  that  of  natural  teeth  and  therefore  deep  epithelial  proliferation  around  implants  easily  occurs1).  Recently,  the  excellent  sterilization  potential  and  irrigation  power  of  functional  waters{the  strongly  acidic  electrolyzed  water(SAEW)  and  the  strongly  alkaline  electrolyzed  water(SALEW)}  has  gotten  a  lot  of  attention  and  its  use  is  widely  increasing2,3,4).  The  purpose  of  this  study  was  to  investigate  the  existence  of  periodontal  pathogenic  bacteria  in  peri-­implant  sulcus  and  effects  of  irrigation  by  functional  waters  (Table  1).
Control  group  Figure  1
Experimental  protocol

Syringe(25ml),
Irrigation  using  saline(100ml)

Table  1    The  features  of  strongly  acidic  electrolyzed  water  (SAEW) 2)  Broad
antibacterial
spectrumNo allergy
No side effectNo residue

Strong virucidal
effectPrepoerative  samples

Sterilized  pledget

Specialized  electrolysis  processing  (0.05％  NaCl  solution)
High  disinfection  effect  of  high  oxidoreduction  potential  (ORP)(pH  2.7,  ORP  1100mV)

Paper  point
Supragingival  plaquewas  removed(including HIV, HBV )Reasonably cheap(240Ɛ/$ only)

Maintains high safety
level

Environment
friendly

Paper  point
Postoperative  samples  Toothbrush(Figure  1-­C)

Peri-­implant  sulcus  (Figure  1-­A)(Figure  1-­D)(Figure  1-­B)

DNA  of  the  periodontal  pathogenic
bacteria  in  the  sample(both  groups  of  paper  point)  was  estimated  by  the  PCR-­invader  method.

Experimental  group

No resistant bacteria

Syringe(25ml),Irrigation  using  SAEW  and  SALEW(100ml)

Ⅱ  Material  and  method：

17  subjects  treated  with  implant  therapy  for  single  tooth  gaps  were  included  in  this  study,  and  randomly  assigned  to  the  experimental  group  and  the  control  group.    The  experimental  group  consisted  of  10  subjects  (age  57±13)  and  the  control  group  consisted  of  seven  subject  (age  69±7)    (Table  2,3)．Supragingival  plaque  on  implants  was  removed  with  sterilized  pledget  and  toothbrush  and  paper  points  were  inserted  at  the  deepest  site  of  the  peri-­implant  sulcus(Figure  1-­A).  These  paper  points  were  used  as  preoperative  samples(Figure  1-­B).  Next,  the  sulcus  was  washed  with  the  functional  water  (100ml)  in  the  experimental  group,  meanwhile  the  sterilized  saline  water  (100ml)  was  applied  in  the  control  group(Figure  1-­C).  Further,  paper  points  were  inserted  at  the  same  site  and  used  as  postoperative  samples(Figure  1-­D).    The  functional  water  was  generated  with  three  cylinder-­type  electrolyte  lonize  water  device  {PH2.7(SAEW),  PH11~12(SALEW),density  of  effective  chlorine;;30ppm}(Figure  2）.    DNA  of  the  periodontal  pathogenic  bacteria  in  the  samples  was  estimated  by  the  PCR-­invader  method{PRISM  7900  Sequence  Detection  System  (  A.Applied.Biosystems)｝．Investigated  periodontal  pathogenic  bacteria  were  Aggregatibacter  actinomycetemcomitans  (Aa),  Porphyromonas  gingivalis  (Pg),  Tannerella  forsythensis  (Tf)  and  Treponema  denticola  (Td).    The  study  design  was  approved  by  the  dental  ethics  committee  of  the  general  implant  research  center  and  informed  consent  was  obtained  from  each  subjects.
Table 2Experimental  group        （Using  strong  alkaline  electrolyzed  water，strong  acid  electrolyzed  water）

Table  3Control  group        （Using  sterilized  saline）

SubjectAgeSexSite

The average
period offunction(month)

A69

male

3641

Straumann SLA
WN10mm-555324K67

B55

female143

Straumann SLA BL
RC10mm-235330LC44female

1242

Straumann SLA
RN3.3 12mm

132111D57

female2253

Straumann SLA
RN3.3 12mm

-22123E63

female3639

Straumann SLA
WN10mm-4424F

61

male

36120

Straumann SLA
WN10mm-411G77

female159

Straumann SLA
RN4.1 12mm

-64H64male

1794

Straumann SLA
WN10mm

-5I

48

female2215

Straumann BL
RC4.1 10mm

J32female

259

Straumann SLA
RN4.1 10mm

ƃ：WKUHHƂ：seven
Average    age：57±13  Implant

Perio-test Value

Probing
depth(mm) M

L

B

D

Boneresorption(mm)

Subject

Age

SexSiteThe average
period offunction(month)Implant

Perio-test Value

Probing
depth(mm) M

L

DB

Boneresorption
(mm)

male

3622

Straumann SLA
WN10mm-52

311060female3560

Straumann SLA
WN10mm-542321M

83

female3576

Straumann SLA
RN4.110mm-4323312N67

female2459

Straumann SLA
RN4.1 10mm-42121143O68male

2441

Straumann SLA
RN4.1 10mm

-511111131P64female

2443

Straumann SLA
RN4.1 10mm

1111103321Q75

male

3676

Straumann SLA
WN10mm-32112243342-434440-3

11110

ƃ：WKUHHƂ：fourAverage    age：69.1±7.6  Figure  2    Three  cylinder-­type  electrolyte  lonize  water  device                                (FINEOXER  Fo-­1000,  FIRSTOCEAN  ltd,                            Yokohama,Japan)

The  average  period  of  function：53.9±19.8  months

The  average  period  of  function：42.5±38.5  months

Ⅲ  Result：In  the  control  group,  the  average  number  of  total  bacteria  count  was  12,900,000  before  the  irrigation  and  2,437,143  after  the  irrigation  (P  >  0.05  ，Figure  3,4).    With  regard  to  the  average  number  of  periodontal  pathogenic  bacteria,  PCR  analysis  showed  no  significant  decrease  respectively  (P  >  0.05,  Figure  5,6,11).  In  the  experimental  group,  the  individually  total bacteria  count  decreased  sharply  (Figure  7).  The  average  number  of  total  bacteria  count  was  12,306,000  before  the  irrigation  and  2480  after  the  irrigation.    There  was  a  significant  decrease  (P  <  0.05,  Figure  8).  Among  four  bacteria,  Pg  decreased  significantly  from  an  average  of  453,333  to  an  average  of  15,090  (P  <  0.05  ,Figure  9).  Tf    decreased  significantly  from  an  average  of  659,871  to  an  average  of  7142  (P  <  0.07  ,Figure  10).  Td  decreased  significantly  from  an  average  of  348,383  to  an  average  of  25,316  (P  <  0.06  ,Figure  12).  

Ⅳ  Discussion                        and  conclusion：
In  this  study,  although  the  subjects  did  not  have  periodontal  diseases,  existence  of  red  complex  was  confirmed  in  peri-­implant  sulcus.  Further,  it  has  been  suggested  that  the  irrigation  with  functional  waters  (SAEW  and  SALEW)  for  peri-­implant  sulcus  is  effective  for  reduction  of  periodontal  pathogenic  bacteria.

Ⅴ  References：    Ito  H,  Enomoto  S,  Kobayashi  K:Electron  microscopic  study  of  the  human  epithelial  attachment.,Bull  Tokyo  Med  Dent  Univ.  1967  Jun;;14(2):267-­77.
１）    Shimizu  Y,  Hurusawa  T:  The  sterilization  action  and  fungal  disinfection  action  of  high    oxidation  potential  water  made  from  electrolysis.  Shika  Journal,  37  :1055-­1060,1993  (in    Japanese)  .
２）    Shimizu  Y:Virucidal  effects  of  electrolyzed  oxidizing  water.  The  Medical  &  Test  Journal  Special  Issue  Jan.1,1994.
３）    INOUE  K，ENDO  T，MIURA  H,  IMAI  S  ,  HANADA  N，SENPUKU  H，  SATO  T  :Comparison  of  Washing  and  Antiseptic  Properties  between  Strong  AlkalineElectrolyzed  Water  and  Alkaline  Cleaning  Agent  in  Dentistry,  Journal  of  The  Japan  Society  for  Oral  Functional  Water,Volume12-­1,2011:40-­41.
４）