Osteogenesis in the healing of the extraction wound

OSTEOGH{ESTS T}T

TI{E

HEAI,ING OF

lHE

Ð(IBACTION ^I,{OU}TD

CIIAU Kaí-Kin _o B.D.S.ïlons ^.

*resis

subnitted.

for the

d.egree

of

MASTER OF ^{DEISTAT SURGEFY}

DEPARTIITE},II¡ OF DEI.ÍIAT, SCÏE1\TCE

UNTVERSITY OF ADE,ATDE

.

1968

l-

TABLE

OF

CO}TTHSTS

PRECTS

DECLAR.LTIO}T

ACKITOI,lLEÐGEMENTS

T¡MRODUCTION

CHAPIER

1:

0STEOGENESTS

IN

BONE REPAIR

Historícal

and.

turrent

Concepts

of

^{Bone Fepair}

A.

Periosteum a¡d. bone

celIs

Page

viii

l_x

0.1

1.1

1.3

1.1+

1.7

1.13

1.15

1.16 1.1?

9

^pages

20 pages

vI

T

ïï

B. Induetion of

eonnective

tissue

Tosfaplasi-a

Matrix

Synthesís and

Mineralization

A.

Osteoblasts a,nd ground substance

B.

Osteoblasts and- collagen

Ì formatíon

0. CalcifÍcation

CITAPIER 2: TTTE TTÏSTOIOGY OF BOI$E FEPATR

f.

Healing

in Fractures of

^Bone

A.

The

histological

ser¿uenee

B.

^Bone fo:mation

C. Cartílage

formation

D. O¡antity of

callus

ff. IÏealing of

Cortical- Defeets

A. Periosteal

and. endosteel calluses

B. Remodelling

^,

C.

Role and.

fate of

the

periosteaL

call-us

GrA]ltffi

3:

^HEALII{G

ï}I

THE EXTRAüIION ^WOUIIID

2.t 2.t 2.I

2.6 2.9 2.L2

2.r5 2.r5

2,U

2 .18

11

20 pages

I.

II.

ITI.

The Sequence

of

^Pepair

A. ïn

dogs

B. ïn rats

C. ïn

rhesus monkeys

D. fn

man

LocaL Factors

Influencing

Healing

Dela¡r

in

liealíng

( t'D4r soeketst')

3.r.

?l

3.1 3.2 3.h

?q

3.8

X.5 pages

CI{AT{TER

I+:

OBJECTS .A}TD }4ETTTODOIOGY

T.

CeLl

Proliferation II.

^Gror¡nd. Substance and

Intercellular

^Fibres

III.

^{New Bone Fomation}

A.

Tetracyclínes

B.

Chl-orazol- Fast ?ínk

CHAFTER

5:

MASENTAI.S A}TD METHOÐ$

I.

Ctroice

of

Experimental- Anirnals

Lr

l+.3

4.6 l+.ro

l+.

fe

h. 16

5. ^l-

5.t

5.3

5.1+

5.8 5.9

6. ^r_

6.3

L11

18 ^pages

9

^pages

90

^pages

IÏ.

TTT.

C$.APÍER

6:

^RESüLTS

Operative Procedures E'xperÍment

al

^Procedures

Table 5.1 Table _5.2

I. Tnterpretation

II. I{ealing ín the

Extra.ction Socket

CHAPTER

7:

^{DTSCUSSÏON ?.1-}

?.1

?.h

7.5 7.6 7.8

7.rt

?.11

"f t¿

7.18 T. t9

7.2Ð '7 ^r;'1

B.r

iv

23 pages

2

pqges

ï.

Healing

in the

Extra.ctíon Socket

A.

Granulation Tissue and

Intereellular

Fibres

B.

Period.ontal Membrane Remnants

C.

Resorptíon

in

Alveol-ar Soeket

D.Osteogenesis

E.

Bone

in the

Soehet

F.

Endosteal Bone

G. Periosteal

Bone

II. Correlation wÍth

Healing

Ín

^{Fractures and.}

Cortical

Defeets

A. Periosteal

Reaction

B.

Endosteal Reaction

C.

^Osseous

vepair of injured

Bone

Ir.

Remodellíng

CTÍAPTER

B:

^SUMMÁ3Y .A}.ID CONCLUSTOSS

APPMMCS

RE¡I'EREISCES

ï. nat Dentition

ïï.

Diet

ïïï.

Instntvrents and.

operative

Proeedureg

ïV.

Drugs

V. Ijltra-rrioLet

I'luores eence micros copy

Vï.

Preparation and. Sectioníng

of

Undeealcifíed. Bone

VIï.

HistoJ-ogical and.'

IIisto-

chemical Techniques

App. i App. iii

¡p,p. v

App.

t¡:iiÍ

x

xií

App. xvi

9

23 pages App

App

28 pages

vt

PREgTS

In

a preJ-ininarry

stuðy, it vas

^formdl

that certaín features in tbe healÍng of the

e:ctractíon wound.

in experi-

mental animals were

inconsÍstent with

eonventioaaL descrip-

tions. In

^ad.dÍtion

to the

formation

of

^{ner¡ bone}

withín

^the

alveol-ar socket, bone was

also

forrned suboeriosteaLly on the buccal

surface.

^Ttre

present investígation is

an extension

of the

aþove study.

A hooded

strain of the

$forwegía,n

rat in rrhÍdr

a

nalclllary

molar was removed. has been

used,

artd

the

healinel proeess

that follor¡ed

^l¡as

stutlied wÍth the

use

of:-

1. Intra-vítaL staining of

^{ner¡ bone}

rríth

(

a) Ae¡ethyl-chlortetraeyeline, (t)

chLorazol

fast pink.

2, Localization of mitotic activíties with

coLchicíne.

3.

^Sone

histo-eherlical

observations

of extra-eeLlular

eorcponents vhieh were also ^{mad.e along}

with routine histologica.i-

technio-ues .

The

results

índ.ícate

thai there is a

close

parallel

between

heating in fractures, ln corbical

clefects

in

^bone

and

in the alveolar extraction

^wound, by

the fonratÍon of

ttperiostealtt

r

ttendostealrf and rrunitingtt

eallus.

vÍí

Hovever,

the

formation

of

buccal

periosteal

_bone

in

the

extraction

wound was a

variable feature

and appeared

to'be

reLated

to the

Þresenee

of inflainnatÍon.

Inf1q¡rmation

Ín the

perioster¡n could.

asise fron

⁽

a) a d:ireet

extension

frm the oral

wound,

or (b) ind.ireetly

across

the alveolar

socket

wal.l.

Ttle

lack of periosteal

bone formation

in the

nasal

and. palataL surfaces

of the

rna:<ilJ.a cannot

be

explaineil.

Ccrrnparison

with repair of

bone

in other sltuations

suggests

that

under

certain

eireumstanees, Ínflamtration

viLl

lead

to periosteal

_bone for¡aation.

Food.

ínpaetion

end sequestration

of necrotic

bone anrL/or

root

fra€ments leacls

to intensive

infl-qr¡rnation

in the

soeket.

lhis intensive

infla¡mnation d.el4yecL wound. heal-ing and. bone fo:¡mation

ín the

socket.

End.osteal bone

fo:mation,

analogous

to the

endosteal

callus in the healÍng of a fracturer

wâs aLso noted

Ín

^some

sneei¡ens.

vÍii

DECLARATIOlV

fhis thesis ís

submitted

in fu]-filnent of the require-

ments

for the

Degree

of

^Master

of

Dental Surgery

in

the

Universit¡¡ of Adelaid.e.

Cand.idature

for the

Degree nas

satisfied by obtaining the

Honours Ðegree

(¡'irst

Class)

of

Bachelor

of

Dental Surgery

in

1965.

Ttris thesis

contains no

material which,

except where due

nention is

made, has been aecepted,

for the

^at¡ard.

of

^any

other

^d.egree

or

d.iplcma

Ín

any

University.

^To

the best of

my knowl-ed.geo

this thesis

contains no

material

previously publishetL

or lrrítten by

another person, exeept where clue

ref erenee has been mad.e

in the text.

Ttre

results of the present investigatÍon

have ^been presented.

in part to the

^meeting

of the

International- Assoeiation

for

^{Dental Researeh}

(Australian Section) tg6l.

CIIAU Kai-Kin

1X

A CKI\TOI'ILÐG$ IEÌ\TTS

The author wishes

to

^record

his

sÍncere

gratitude to his

supervÍsoro

Dr. J.A.

^{Cran, Reader}

in

0ra1 Pathology and 0ra1 Surgery,

the University of

Aclelaide,

for the o¡porbunities

^he has made

ar¡ailable for thís

study and

for his

^{constant aclvíce}

'

eneouragement and patience.

To

the staff of the

Histo-pathology

Laboratory, of

the

Divísion of Oral

Pathology ar¡d

Oral

Surgery,

Ðd in parbicular to l¡fr.

D.E. Smale,

the

autlror

is very

^mueh indebted

for their

eo-operatíon ^and

technieal assistance.

To these persons,

&d to

^Miss

K.

Shertoek, who prepared

the

photogfaphic

illustrationst the author

also wishes

to

record

hís

appreciation.

Thanks

are also

due

to Prof. E.

^Storeyo

the

^UníversÍty

of

Melbourne,

for the

opportunit-r¡

for learning the

^techniques

of intra-vital

bone

staíning

and.

the preparation for

sectíoning

of

und.eealcified. tissues ^.

Lastly, the.author

^wishes

to

^{thank Miss}

E.B.

^{James and}

Mrs. R.

Scantlebury

for their

care taken

in the typing of

this

manuscript.

INTRODUCTION

Although r,¡ound

healing is the basis of

surgíca1 seience

the intricate

meehanisms

of this essential

process are

by

no means

und.erstood.. It is not until recently that

atte¡rpts have been made

to correlate the histological,

bioetremical and

physical

aspects

of

wound healing.

Becar:se

of its

hard.

nhysieal nature,

bone

tissue

poses

special

problerns whieh

have, for a long

tÍme, d.efÍed. the

application of

experimental methods r,rhich have

yielded results in investigations in healing of soft tissues.

Under nor:mal

conditions,

wounds

of the

mouth

heal ranídly

and

without

measurable systemic

effects. Relatively for detailed investigations, eÍther clinical or

experi:nental, have been concerned

with this

subject.

DiscrepancÍes

in the literature

regard.ing

the

histogenesís

of various tissues

associ.a,ted-

with

al-veolar socket wound

healíng

pronpted, a prelímínar¡r

investigation

by

the

author.

As an outcorne

of this, certain features in the healing of

^the

erçtraetion wound ^rn¡ere noted. r,rhich called.

for further

inves-

tigation.

The present

investigation is

an

attanpt at ap¡lying

^some

teehniques

of studying ealcified. tÍssues to the healing of

o .2.

extraetion wor¡nd,s. At the

sane

time, the

^r¡ound has al.so been

studied frcn the histological

and. histochenícaL ângle.

An

attenpt

uas made

to

^ecrnpare and eorrel-ate

the healíng of this intra-oral-

wound

wíth the healing of a fracture in

long bones and-

healing of eortieal

clefects

in

^bone.

fhe effect of

feeding mad.d.er

root

on

the

^bones

of pigs, first

observed duríng

the niddle of the

eigbteenth centurXr, gave anatcmists

of that period a tool for follorÍng

the development

of

^and"

repair

^processes

in bone. ïhe

basie observations

of early

workers

are stil1 vaIid.,

and

constitute the

formd.atíon

of eellul-ar

bone ¡ürysÍology as

it exists

today.

^The teehniques

of intra-r¡:itaI staíning of

bone using varÍor¡s markers

are

refinements

of the

same

principle

and

these fomr

the first part of the

present

investigation.

ïn

recent years ^rnan¡r methods

of tagging

and label-ling ceI1s have beccme

availeble with the

use

of

::ad-io-aetive

isotopes

and. have

afforded

a meons

of

studying

the origin

^and

fate of

eelluJ.ar ccrrponents

in

tíssues

"

Less aecurate ^and.

sorplistieated. methods have aJ-so been avaíIab1e which enerble a

lÍnited insight into cellular activities.

One

of

^these

latter

methods,

the

use

of coldri.cine,

has been

applied in the

present

investÍgation to

observe

the ¡ritotLc activities of

various

eellular

eomponents

in

bone

repair.

0.3.

Ttre rol-e

of the

ground substances and.

the extra- eellular

cæponents have been emphasised.

in recent

stud.ies on wound

healing.

Histoehenical raethods

afford

a means

of íctentifying

varlor.s bioehemical ccmponents present

Ín

the

healing

r,¡ouncl and.

fóm a

subsid.iar¡r approach

to the

present

study of

^osseous

repair.

ïn the first seetion of this thesís, the

rel-evant

Literature

on

various

aspects

of

bone

repaír

and. osteo- genesis

are revie'wed..

The

theoretical basis of

^methotts

used

in the

present

investigation is d-iscussed.

The seeond.

seetion

reporbs

the

^methocts and.

findings.