The influences of Perftoran on the local circulation and osteoreparation for the experimental high-energetic diaphysis fractures depend on the method of introductions

8/14/2019 The influences of Perftoran on the local circulation and osteoreparation for the experimental high-energetic diaphy

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Perftoran, experimental fracture, microcirculation, laser dopplerflowmeter, osteogenesis

G. M. Dubrovin, A. V. Ivanov, O. V. Yakovleva, Chen Chi Hsiang,D. A. Ivanov

The influences of Perftoran on the local circulation andosteoreparation for the experimental high-energetic diaphysis

fractures depend on the method of introductionsKursk State Medical University, Kursk

There are increased portions of high-energetic injury in generalstructual trauma in the every year [3]. The frequency of development isslowly in the consolidation with prosthetic joints in ones according to thedifferent authors , there are attaint to the amount in 33.2% , ones in givendata from the consequences of high-energetic fractures in long inferiorextrmities[1,2,16].

With given evidents of the important stages in speed and quality ofconsolidation in fracture, is depending in the situation upon thehemodynamics of injurious extrmities[2,3,612,13]. In the one so called

priority tendency in the development of modern traumatology is theelaboration of the well-based pathogenetic methods which is the mostsignificant, to acquired by means of restore the adquate tissues perfusionthrough the stimulation in osteoreparation[6,10]. With these views of

points were to represent the application of Perftoran as the majority ofinterests in study regeneration in traumatology.

Perftoran- the blood substitute with air-transportive function and inwhich there are possessing hemodynamics, rheologics,membrane-satblizings,cardio-proectorings,diuretics and the properties of sorption[5].The application of Perftoran which be able to afford as, to enhance thecontent of oxygen transpotation, to standardize the utilization of tissuesoxygen and to create the pre-requisite for relive the hypoxia[8,9,15]. ThePerfluorocarbon emulsion has possesses the resources of formed so called

perfluorophages. These Perfluorophges are to preserve the functionlactivity and to take a part in the process of regeneration till to healing thewound, by means of this, it given the process of regeneration to beaccelerated. With the revealing phenomenon a steady increases of the

partial pressure of the oxygen in the wounded tissues at the zone ofPerftorans which were presented, there are appeared to be assured of thetissues oxygenation, and to stimulating the reparation and theregeneration, and to preventing the pathological fibro- andcollagenesis[4]. There are basic mechanisms in the reduction of thehypoxic tissues, to implement in the oxygen expenses, by the gradients of

partial pressure makes the solution of Perftoran to transitory in tissues, beexpressed to reduce the frequency of infectious complications and to


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increase quantity of viable cells in the area of traumatic destruction ofbones[11,14].

In the present time, the aims of using Perftoran are to keep thetissues viabilities in the zones of fire-arm shotting fractures[11], plus

with treatment in opened fractures by using Ilizarov method[15] and soon.. However abrupt infrigement of regional blood current is not allow toeliminate the local hypoxia in the place of fracture it is that we wish thegas exchange not be processing and, neither intravenous introduction norfor Perftoran perfusion in the zones of fractures.

The aim of our investigation to study the influences of thedynamics in osteoreparation under the experimental high-energeticfracutres on the long tubular bones for different methods of Perftroanintroductions.

Materials and methods

The experimental investigation is carried out on the 100 male rats(Wistar), and the weight of each is about 220-260 grams. The operativeintervention and morbid manipulation for animals all were conducted tonarcotic in accordance with The regulations of humane treatment withlaboratory animals. In the during operation and after opertation periodsrats were all applied with antibiotics therapy.

First stage in the experiment were modelization high-energeticfractures at the upper 3rd femoral bones of right posterior extremity: it isafter the transverse osteotomy was conducted with the removal of

periosteum and endosteum, at 0.5 cm. from the line of fractures, on thedistal and proximal fragments, plus with mashed off soft tissues in thezone of fractures with the help of clamp. Further was conducted with theintramedullary osteosynthesis slim drill, then the wound be suturedtightly.

The animals after modelization in fractures were allocated into fourgroups: one control group and three trial groups, in each group there is 25rats (Tab.1).Table 1 Scheme of experiment


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In the 1st control group the preparation for animals were not beintroduced.

In the 2nd group at 7,10, 13 days after operation in intramedullary

with introduced of Perftoran (Open Joint stock compny, SPFPERFTORN, Pushchino city. Moscow) at the rate of 1 ml. per 100 g.weigt of animal (in average 2.4 ml. Perftroan, dosage selected by meansof experiment). In the 3rd group animals in these same periods introducedintraperitoneum 3 ml. in Perftoran.

In 4th group upon the indication days, Perftoran is in injection atquantity of 2 ml. introduced in interfragments zone.

Using the amount of 5 rats from each every groups on theexperiment at 15th,21st,30th, 45th, and 60th days of correspond withafter operation (it is in introduction to carry out the approximatemeasured doage of Chloral hydrate intoduced into intraperitonium).There are animals for all groups were in the fact of hatching for trials tocarry out, the during life times investigation of microcirculation.

The method of control standard microcirculation in the zone ofinjury were chose in Laser Doppler Flowmeter (LDF), which candefinites the stages of blood current compensation and was so called oneof the basic methods for this aspect on the present days, it is able toaffords the analize of microcirculation in the realistic scale times[7,17].The dynamics of indicators on the all groups were to be estimated by the

help of the apparatus in model of Biopac MP-100 with the needle likesensor in TSD-144 type. It is to computes the intergral exponent of

Groups Scheme of experiment: quantity of animals, periods of introduction in

preparation and conclusion of animals from trials

Modelizationoffractures

Introduction of

preparation

conclusion of animals from trials

7th

days

10th

days

13th

days

15th

days

21st

days

30th

days

45th

days

60th

days

1st Controls

25 25 25 5 5 5 5 5

2nd Introduction of Perftoran in intraosseous25 25 25 5 5 5 5 5

3rd Introduction of Perftoran in intraperitoneum

25 25 25 5 5 5 5 54th Introduction of Perftoran in interfragments zone25 25 25 5 5 5 5 5


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microcirculation in the marked by BPU (Blood Perfusion Unit) andwhich can characterizes the perfusion of tissues. The investigation of

blood current in the capillaries of the tissues in the zones of injury candivide into 3 ground levels: it is the interfragments callus (the tissue

reside between fragments), the periosteum of distal fragment at thedistance of 0.5 cm. from the line of fracture, the muscular tissue in thearea of fracture. The recording and processing of data to implement bythe software program AcqKnowledge-38. The digital data to processesinto statistics with the help of analytical package of MS Excel.

For the morphological investigation is to excises the fragments onthe extrmities, that is including the zones of experiment influences onwith the surrounding tissues. After the decalcification of the tissues, withfurther processing in histological preparation and utilizing the micotometo slicing the tissues into 10-12 micro meter dying the preparations withthe methods of Van Gieson stain, Mallory and Hemotoxylin-Eosin. Themorphometry with the conduct help of software program Image J toconverses into the statistical data.

The consolidation of fracture was evaluated by roentgenologicalmethod (x-ray ) to estimate for data using.

ResultsThere are all of the animals which were participants in the

experiment, the wound were healed with initial tension ( without pus incleaned wound ). The results of the study in the microcirculation of in

different periods of observations were reflected on the diagrams.


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Picture 1. The Chang of microcirculation in the interfragments.

Y-axis: BPU (Blood perfusion unit), X-axis: Days

The denotes of colors:

Deep blue: 1st group (------) (control group)

Light blue: 2nd group (____) (Perftoran intraosseous)

Light green: 3rd group (____) (Perftoran interperitoneum)

Red: 4th group (___) (Perftoran interfractures zones)

66,66 72,12

177,64 180,96 182,38

94,22

73,54

59,36

227,77

76,32

92,2 89,0467,36

64,89

115,34 118,11

211,98

0

50

100

150

200

250

15 21 30 45 60

BPU

1 () 2 ( /)

3 ( /) 4 ( . )


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Picture 2. The Chang of microcirculation in the periosteum.Y-axis: BPU (Blood perfusion unit), X-axis: Days

The denotes of colors:

Deep blue: 1st group (------) (control group)




91

77

107

167

106

46

129

202

304

83

113

124

60

107

94 92

203

0

50

100

150

200

250

300

350

15 21 30 45 60

BPU

1 () 2 ( /)

3 ( /) 4 ( . )


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Picture 3. The Chang of microcirculation in the muscles.Y-axis: BPU (Blood perfusion unit), X-axis: Days

The denotes of colors:Deep blue: 1st group (------) (control group)




323

666

1009

418

735

1085

607554781

1157

1060

1240

876

1515

586527

641

0

200

400

600

800

1000

1200

1400

1600

15 21 30 45 60

BPU

1 () 2 ( /)

3 ( /) 4 ( . )


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In the tissues of interfractures zones revealed the evidence of themicrocirculation (BPU) at 15 days of the experiment by the statistics in

authenticly higher from the series of with intramedullary (227.9883.3)introduction of Perftoran and the directly introduction of Perftoran intothe interfractures zones (211.9844.9) in compared with the others groups(pic.1).At the 21st days after modelization into fractures, it is records fromthe statistics in authenticly at 2-2.5 times higher in the 2nd group becompared with the others groups, it is the evidence of the circulation fromthe formation of callous. Moreover, from above of the statistics that thisexceeding keeps on till 45 days time. The BPU of the 4 th group (theintroduction of Perftoran into the interfractures) beginning at the 21 daystill the ends of experiment which was lowering in the value,it is donthave authentic distinction with first and 3rd groups. In the point ofinterfragment callous the first and third both groups on the authenticly ofthe statitics does not had distinct evidence in with the micrcirculation ofall the extent of experiment.

The study in the microcirculation at the periosteum gave theevidence of the maximun BPU (20384.6) value from the interfracturescoexist with the introduction of Perftoran at 15 days (pic. 2). However,from the 21 days and till the ends of experiment proved themicrocirculation on the 2nd group (intraosseous introduction of Perftoran)

had excceded BPU than other groups, moreover, at 21 days thisexceeding had have statistical authentic signaficance (30454.8). In thecontrol and 3rd groups had proved that the microcirculation in thestatistics does not have differences and be with the least meanings.

In the muscular tissues, by the data which were gained from theLDF, the better blood circulation by the observed on the intraperitoneumwith method of introduction of Perftoran at 30 days and 45 days (pic.3).Inthe other groups revealed that BPU were lower and essential distinctions

between each others were not revealed.For all the investigating groups till at the 60 days in the

experiment, proved there are the microcirculation on the intrafracturestissues, the periosteum and the muscles all practically made theequalization in statistics in BPU and not had the differences amongthemsleves too.

The analysis of roentgenogram (X-ray) bring out from the trial ofanimals proved as follows: in the control group have had one of the casesin consolidation of fracture that begun at the 45 days and the other case isat the 60 days. In the other cases, it is developed the artificial joint ( pic.4).


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Picture 4. The roentgenogram (x-ray) and microphotography of fracture.

Control gruop (45 days): artificial joint; The presence of cartilage tissues and

cavity of different volumes with biased to merging in the intrafracture zone

(Hemotoxylin-Eosion, zoom in 100).

In the 2nd group there are the consolidation were observed on, it isfrom the one of the animals at 30 days, from the fours at the 45 days andfrom the fours by the introduction of trials at the 60 days ( pic. 5). Onlythere is one case in the result of migration of drill was to turn out in theartificial joint. The similar results gained from the 4th group: there are 2

cases in consolidations at the 45 days and other 2 cases at the 60 days.


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Picture 5. The roentgenogram (x-ray) and microphotography of fracture.

The grup of investigation 2 (45 days): the consolidation of fracture;

The formed callus from the fibrous tissues; (dyeing, Hemotoxylin-Eosion,

zoom in 100).

From the acquisitions of data by the roentgenographic investigationthere are conform to with the morphological investigations of the

preparations ( pic. 4, 5). There is revealing that the intramedullary

introduction of Perftoran have the essential to hasten the organization of bony calluses. In such a way, more narrowly at the 30 days aftermodelization of fracture, the thickness of calluses was to formed at thequantity of 0.170.08 mm, it is the statistics in which the authenticly todistinguish from the control groups and the series of experiment with theintramuscular introduction of Perftoran. At the 45 days in the most

preparations of the interfractures spaces were occupied by coarse fibrousconnective tissues with the islets of cartilage tissues, which is observed toin the formation of the trabecular bones with morphological indicationsof ossification.

DiscussionThere are many authors to mentioned about, that owing to the

broad spectrum of the activity of Perftoran, which is sufficiently witheffective for the intravascular infusion and also the same with the localintroduction (applications)[11,14,15]. But these methods of intrduction of

preparations does not solve the problems of local hypoxia, which isoccurred especially in the bone tissues due to the high-energetic fracture.

The analysis of dopplergram revealed the important increasementevidences in microcirculation of BPU at the interfractures tissues with the

intraosseous introduction of Perftoran as practically along on the whole


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experitment. In the begun with the 21 days to the 45 days of after theinitial experiment there were had 2-2.5 times in the increasement of thelevel of the microcirculation in the develop of intramedullary callus inthe compare with the other groups. Ther is elevated evidences in

microcirculation connected with the increasement of the amount offunctionl capillaries in the intrefragment callus, and in this period goingon the most intensive callus formation, it is as same as the conformation

by the hitologics and roentgenologics investigations. This is may toexplains why there are exist Perftoran as follows, it is that intraosseousintroduction at the distal metaphysis Perftoran can partially found directlyinside the intraosseous vessels, it is partially to found Perftoranexpression at the interfracture space, that it is allowed to lower down thehypoxia in the place of fracture and it is in some cases directly inside the

bone tissues.There are autheticly at most of the lower evidences in BPU at the

direct introduction of Perftroan in the interfracture zone, but for all thelower ones in BPU there have exceeds more lower evidences as theintroduction of Perftoran in the intraperitoneum (it is as similar as to theintravenous introduction) and as for the control groups both. This toindicated that the injection of the introduction of preparation in theinjured tissues with infringed on the blood current will not allow in thefull value to realize its property, however it is more by preference than theintravenous passage to introduction of Perftoran. The intravenous

introduction of Perftoran not be able to afford to have an influence at thelocal perfusion in the tissues of the zone of high-energetic injury: thereare evidences in BPU in authenticly not to distinguish from the controlgroups.

Another changes in the microcirculation for the measurement at thenon- damage periosteum and muscles in the zone of fracture. The highestmeanings of BPU for the interfractures of introduction of Perftoran atwounds in times of 15 days speak about has possbility to improves themicroculation in the non-damage soft tissues with on the direct contact ofPerftoran. The intravenous introduction of Perftoran is named as theoptimal for the improvement of microcirculation in the structual non-damage muscles in the zone of fracture.The more lower evidences ofcirculation in the non-damage periosteum and muscles of the intraosseousof the introduction of Perftoran to testify about, that for the given methodof introduction of preparation not act on the the relatively well tissues.The investigation proved that the intraosseous method of the introductionof Perftoran to be able to afford to raise its concentration at aiming theischemic tissues at the zone of fracture.

The stated changes of microcirculation in all the groups straight to

conform with the dynamics of osteoreparation. The revelation about thatin the intraosseous utilization of Perftoran, which maximum increased the


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amount of active functional elements of the microcirculative channels,and these are provide for the conditions for active of osteorepapration.Only the intraosseous intorduction of Perftoran bolding in the most casesto get union of fracture. About for all the remains groups were to

prevailed in the development of the artificial joint.Thus, the carrying out of the investigation were confirmed that the

capacity of Perftoran to assure the early timing in more accelerate than inthe control groups of the level of the microcirculation in the zone offracture, that to assist of the strengthening of the osteogenesis. Theintraosseous introduction of Perftoran for the treatment of the high-energetic bone wounds were possessed advantages than the intravenousand local applications, it is supply the optimal oxygenation in injuredtissues in the zone of fracture, it stimulates reparative processes in the

bone tissues and to impedes the development in the amount of artificaljoints.

Conclusions1. The formation of bone calluses was depending on the

conditions of microcirculation. The high-energetic fracture it causes theimprigement of the local blood circulation, hypoxia, and as the result ofobserved cause the indirect osteogenesis. In the control series ofexpeiment were prevailed in the retarded union, the non-connectingfracture and the artificial joints.

2. The intravenous and local applications of Perftoran not

to afford the important to influence on the local hypoxia andmicrocirculation in the zone of fracture because of the sharpimpringement of the local blood current in the injurious tissues.

3. The registration of increase in BPU in the group withintramedullary method of the introduction of Perftoran try to prove, thatnamely this method is apparently with optimum for the high-energeticfractures. The intraosseous introduction of Perftoran optimal localmicrocirculation on the damaged tissues in the zone of fracture, itstimulates reparative processes in bones and impedes the development ofartificial joints.

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Documents

The influences of Perftoran on the local circulation and osteoreparation for the experimental high-energetic diaphysis fractures depend on the method of introductions