Pi is 0889540609009457

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REVIEW ARTICLE Miniscrews in orthodontic treatment: Review and analysis of published clinical trials Adriano G. Crismani, a Michael H. Bertl, b Ales ˇ G. C ˇ elar, a Hans-Peter Bantleon, a and Charles J. Burstone c Vienna, Austria, and Farmington, Conn Introduction: A systematic review of effects related to patient, screw, surgery, and loading on the stability of miniscrews was conducted. Methods: Reports of clinical trials published before September 2007 with at least 30 miniscrews were reviewed. Parameters examined were patient sex and age, location and method of screw placement, screw length and diameter, time, and amount of loading. Results: Fourteen clinical trials included 452 patients and 1519 screws. The mean overall success rate was 83.8% 6 7.4%. Patient sex showed no sig- nificant differences. In terms of age, 1 of 5 studies with patients over 30 years of age showed a significant dif- ference (P \0.05). Screw diameters of 1 to 1.1 mm yielded significantly lower success rates than those of 1.5 to 2.3 mm. One study reported significantly lower success rates for 6-mm vs 8-mm long miniscrews (72% vs 90%). Screw placement with or without a surgical flap showed contradictory results between studies. Three studies showed significantly higher success rates for maxillary than for mandibular screws. Loading and heal- ing period were not significant in the miniscrews’ success rates. Conclusions: All 14 articles described suc- cess rates sufficient for orthodontic treatment. Placement protocols varied markedly. Screws under 8 mm in length and 1.2 mm in diameter should be avoided. Immediate or early loading up to 200 cN was adequate and showed no significant influence on screw stability. (Am J Orthod Dentofacial Orthop 2010;137:108-13) I n 1997, Kanomi 1 first mentioned a temporarily placed miniscrew for orthodontic anchorage. The following years brought more refined screw de- signs. 2 Miniscrews have now become established ortho- dontic anchorage aids, with diameters of 1 to 2.3 mm and lengths of 4 to 21 mm. 3-16 Nevertheless, many case reports and only a few comprehensive studies have been published on orthodontic miniscrews. The ar- ticles promised bright treatment prospects but often lacked evidence-based results. 17,18 Hence, studies on screw design as well as surgical and orthodontic treat- ment procedures are much needed. The aims of this re- view were to analyze the reported success rates of miniscrews and to define guidelines for their selection and application. MATERIAL AND METHODS A Medline search was conducted with 2 search-term combinations: [screw orthodontic] and [implant ortho- dontic]. The terms were chosen generally, since there has been little conformity on the nomenclature. 19 A set of criteria was defined to subsequently filter the re- sulting articles: (1) studies on orthodontic mini-im- plants or miniscrews published in either English or German, (2) human clinical trials, (3) no case reports or case series, (4) no studies with fewer than 30 mini- screws, and (5) additional data on factors related to the patient, miniscrew, surgery, and loading available for correlation with the miniscrews’ success rates. References in the criteria-matching results were searched for additional articles. When available, data were extracted that correlated with the miniscrews’ success rate: patient sex and age, screw length and diameter, method and location of placement, time, and amount of loading. The statistical analyses of these uniformly formatted data and their graphic representation were made by us- ing SPSS software (version 13 for Mac OS X, SPSS, Chicago, Ill). RESULTS As of September 2007, the MedLine search with the terms [screw orthodontic] and [implant orthodontic] returned 734 results. Of those, 14 articles matched all criteria and were considered (Table I). a Professor, Department of Orthodontics, School of Dentistry, Medical Univer- sity of Vienna, Vienna, Austria. b Resident, Department of Orthodontics, School of Dentistry, Medical University of Vienna, Vienna, Austria. c Professor, Division of Orthodontics, School of Dental Medicine, University of Connecticut, Farmington. The authors report no commercial, financial, or propriety interest in the products or companies described in this article. Reprint requests to: Adriano G. Crismani, University Clinic of Orthodontics, Department of Dentistry and Maxillofacial Surgery, Medical University of Innsbruck, Innsbruck, Austria; e-mail, [email protected]. Submitted, December 2007; revised and accepted, January 2008. 0889-5406/$36.00 Copyright Ó 2010 by the American Association of Orthodontists. doi:10.1016/j.ajodo.2008.01.027 108

description

ortho

Transcript of Pi is 0889540609009457

  • REVIEWARTICLE

    Miniscrews in orthodonticanalysis of published clinical trials

    Adriano G. Crismani,a Michael H. Bertl,b Ales G. Celar,a H

    Vienna, Austria, and Farmington, Conn

    Introduction: A systematic review of effects related to patienminiscrews was conducted.Methods:Reports of clinical trials30 miniscrews were reviewed. Parameters examined were patplacement, screw length and diameter, time, and amount of lo

    ratetientd sigs ratoweillarys ratet proiateOrth

    has been little conformity on the nomenclature.19 Aset of criteria was defined to subsequently filter the re-

    ini-im-lish orreports0 mini-lated tovailablees.ts were

    rrelatednd age,tion of

    Chicago, Ill).

    returned 734 results. Of those, 14 articles matched allcriteria and were considered (Table I).

    sity of Vienna, Vienna, Austria.

    Connecticut, Farmington.

    0889-5406/$36.00

    Copyright 2010 by the American Association of Orthodontists.doi:10.1016/j.ajodo.2008.01.027108RESULTS

    As of September 2007, the MedLine search with theterms [screw orthodontic] and [implant orthodontic]

    The authors report no commercial, financial, or propriety interest in the products

    or companies described in this article.

    Reprint requests to: Adriano G. Crismani, University Clinic of Orthodontics,

    Department of Dentistry and Maxillofacial Surgery, Medical University of

    Innsbruck, Innsbruck, Austria; e-mail, [email protected].

    Submitted, December 2007; revised and accepted, January 2008.The statistical analyses of these uniformly formatteddata and their graphic representation were made by us-ing SPSS software (version 13 for Mac OS X, SPSS,

    bResident, Department of Orthodontics, School of Dentistry,Medical University

    of Vienna, Vienna, Austria.cProfessor, Division of Orthodontics, School of Dental Medicine, University ofcase reports and only a few comprehensive studieshave been published on orthodontic miniscrews. The ar-ticles promised bright treatment prospects but oftenlacked evidence-based results.17,18 Hence, studies onscrew design as well as surgical and orthodontic treat-ment procedures are much needed. The aims of this re-view were to analyze the reported success rates ofminiscrews and to define guidelines for their selectionand application.

    sulting articles: (1) studies on orthodontic mplants or miniscrews published in either EngGerman, (2) human clinical trials, (3) no caseor case series, (4) no studies with fewer than 3screws, and (5) additional data on factors rethe patient, miniscrew, surgery, and loading afor correlation with the miniscrews success rat

    References in the criteria-matching resulsearched for additional articles.

    When available, data were extracted that cowith the miniscrews success rate: patient sex ascrew length and diameter, method and locaplacement, time, and amount of loading.aProfessor, Department of Orthodontics, School of Dentistry, Medical Univer-452 patients and 1519 screws. Themean overall successnificant differences. In terms of age, 1 of 5 studies with paference (P\0.05). Screw diameters of 1 to 1.1 mm yieldeto 2.3 mm. One study reported significantly lower succes90%). Screw placement with or without a surgical flap shstudies showed significantly higher success rates for maxing period were not significant in the miniscrews succescess rates sufficient for orthodontic treatment. Placemenlength and 1.2 mm in diameter should be avoided. Immedshowed no significant influence on screw stability. (Am J

    In 1997, Kanomi1 first mentioned a temporarilyplaced miniscrew for orthodontic anchorage. Thefollowing years brought more refined screw de-

    signs.2 Miniscrews have now become established ortho-dontic anchorage aids, with diameters of 1 to 2.3 mmand lengths of 4 to 21 mm.3-16 Nevertheless, manyans-Peter Bantleon,a and Charles J. Burstonec

    t, screw, surgery, and loading on the stability ofpublished before September 2007 with at leastient sex and age, location and method of screwading. Results: Fourteen clinical trials includedwas 83.8%6 7.4%. Patient sex showed no sig-s over 30 years of age showed a significant dif-nificantly lower success rates than those of 1.5es for 6-mm vs 8-mm long miniscrews (72% vsd contradictory results between studies. Threethan for mandibular screws. Loading and heal-s. Conclusions: All 14 articles described suc-tocols varied markedly. Screws under 8 mm inor early loading up to 200 cN was adequate andod Dentofacial Orthop 2010;137:108-13)

    MATERIAL AND METHODS

    AMedline search was conducted with 2 search-termcombinations: [screw orthodontic] and [implant ortho-dontic]. The terms were chosen generally, since theretreatment: Review and

  • screw

    d with failure of mini-implants Int J Maxillofac Implants 2004

    ntic forces? Am J Orthod Dentofacial Orthop 2004

    orthodontic anchorage J Orofac Orthop 2004

    ew implant anchorage Angle Orthod 2005

    ng an orthodontic mini-implant Clin Oral Implants Res 2006

    mplants used as orthodontic Am J Orthod Dentofacial Orthop 2006

    rage J Oral Maxillofac Surg 2006

    c anchorage Int J Oral Maxillofac Surg 2006

    traction: a pilot study Am J Orthod Dentofacial Orthop 2006

    ic anchorage: success rates and Am J Orthod Dentofacial Orthop 2007

    for orthodontic anchorage: Clin Oral Implants Res 2007

    lescents Int J Oral Maxillofac Surg 2007

    e different orthodontic skeletal Clin Oral Implants Res 2007

    Table II. Overall success rate

    Authors Screws (n) Patients (n) Success rate (%)

    Miyawaki et al3 134 44 77.8

    Cheng et al4 92 44* 91.3

    American Journal of Orthodontics and Dentofacial Orthopedics Crismani et al 109The overall success rates were available in all 14 ar-ticles and ranged from 59.4% to 100%. The meansuccess rate for all 14 studies averaged 83.6% 610.2%.Weighted by the number of screws in each study,the mean success rate was 83.8% 6 7.4%. The number

    Table I. Articles reviewed

    Authors Title

    Miyawaki et al3 Factors associated with the stability of titanium

    region for orthodontic anchorage

    Cheng et al 4 A prospective study of the risk factors associate

    used for orthodontic anchorage

    Liou et al 5 Do miniscrews remain stationary under orthodo

    Fritz et al 6 Clinical suitability of titanium microscrews for

    preliminary experiences

    Park et al 7 Group distal movement of teeth using microscr

    Motoyoshi et al 8 Recommended placement torque when tighteni

    Park et al 9 Factors affecting the clinical success of screw i

    anchorage

    Chen et al 10 The use of microimplants in orthodontic ancho

    Tseng et al 11 The application of mini-implants for orthodonti

    Herman et al 12 Mini-implant anchorage for maxillary canine re

    Kuroda et al 13 Clinical use of miniscrew implants as orthodont

    postoperative discomfort

    Wiechmann et al 14 Success rate of mini- and micro-implants used

    a prospective clinical study

    Motoyoshi et al 15 Application of orthodontic mini-implants in ado

    Chen et al16 A retrospective analysis of the failure rate of thre

    anchorage systems

    Volume 137, Number 1of treated patients ranged from 13 to 129, with the num-ber of miniscrews from 30 to 273. Overall, the analyzeddata comprised 452 patients treated with 1519 screws(Table II).

    The success rate of the miniscrews was broken downby sex in 6 studies3,7-9,13,16 and by age in 4 studies.3,7,13,16

    This did not include the findings of Liou et al,5 whotreated only female patients of 1 age group. No statisti-cally significant findings in terms of patient sex were ob-served in any studies, but Chen et al16 found significantlygreater success in patients over 30 years of age (Table III).

    The data for screw length and diameter were givenin all included studies. Length ranged from 4 to 21mm and diameter from 1 to 2.3 mm. Nine studies re-ported success rates for screws of either differentlengths or different diameters.3,4,7,9-11,13,14,16 Three ofthem reported statistically significant findings(P\0.05). Miyawaki et al3 concluded that their 1-mmthick screw performed significantly worse than thosewith diameters of 1.5 and 2.3 mm. Similarly, Wiech-mann et al14 reported worse results for 1.1-mm thickscrews than for 1.6-mm ones. In the study of Chen etal,10 the success rate of 6-mm long screws was signifi-cantly lower than that of 8-mm long ones (P\0.05)(Table IV).Journal Year

    s placed in the posterior Am J Orthod Dentofacial Orthop 2003The method and the location of screw placementwere described in all studies. All authors but Miyawakiet al3 used pilot drilling before placing the screw. Bothflap and flapless surgery were performed. Fourstudies4,5,7,9 used a mucoperiosteal flap, and 7 stud-ies6,8,10,11,14-16 used a flapless method. In the remaining3 studies, both flap and flapless surgery were performed,

    Liou et al5 32 16 100

    Fritz et al6 36 17 70.0

    Park et al7 (2005) 30 13 90.0

    Motoyoshi et al8

    (2006)

    124 41 85.5

    Park et al9 (2006) 227 87 91.6

    Chen et al10 (2006) 59 29 84.7

    Tseng et al11 45 25 91.1

    Herman et al12 49 16 59.4

    Kuroda et al13 116 58 86.2

    Wiechmann et al14 133 49 76.7

    Motoyoshi et al15

    (2007)

    169 57 85.2

    Chen et al16 (2007) 273 129* 81.0

    Total 1519 452 Mean: 83.6 6 10.2

    Mean weighted by

    number of screws:

    83.8 6 7.4

    *P\0.05.

  • 7,10,11,13

    20 y

    80.

    83.

    92.

    78.

    78.

    110 Crismani et al American Journal of Orthodontics and Dentofacial Orthopedicsand the corresponding success rates were exam-ined.3,12,13 Although Miyawaki et al3 and Kuroda etal13 reported slightly higher success rates for flaplessplacement, the results of Herman et al12 were consider-ably better with a flap (Fig).

    When information was available, 958 screws wereused in the maxilla with a mean success rate 89.0% 610.0%. In the mandible, 508 screws averaged a successrate of 79.6% 6 8.7%. Weighted by the number ofscrews in each location, the mean success rates were87.9% 6 7.6% for the maxilla and 80.4% 6 8.5% forthe mandible. In 3 studies, the differences between themaxilla and the mandible were significant (P \0.05)in favor of the maxilla (Table V).4,9,16

    Three studies explored the differences between8,9,15

    Table III. Success rates (%) by sex and age

    Authors Male Female \

    Miyawaki et al3 80 84.7

    Cheng et al4

    Liou et al5 100

    Fritz et al6

    Park et al7 (2005) 91.7 88.9

    Motoyoshi et al8 (2006) 90.0 85.1

    Park et al9 (2006) 88.8 93.5

    Chen et al10 (2006)

    Tseng et al11

    Herman et al12

    Kuroda et al13 85.7 88.9

    Wiechmann et al14

    Motoyoshi et al15 (2007)

    Chen et al16 (2007) 84.4 85.4

    *P\0.05.screws placed in the patients left or right side.There was a significant finding (P\0.05) by Park etal9 in favor of the left side. The success rate for theleft side was also higher in the studies by Motoyoshiet al,8,15 although not significantly different.

    The loading variables under evaluation were latencyperiod (time from placement to first loading), overalltime of loading, and force applied to the screws.

    Data on the time until loading were available in 13studies and ranged from immediate loading (#48hours20)to 149 days.Most screwswere thereby incorporated in theorthodonticmechanics, according to the definitions for im-mediate and early loading of implants.20 Four studies pro-vided success rates for different healing periods.3,13,15,16

    Motoyoshi et al15 reported significant differences betweenthe adolescent early-loadgroupand the late-load groupbutalso for the adult early-load group (P\0.05).

    The amount of loading, available in 13 studies, rangedfrom 50 to 400 cN. Kuroda et al13 analyzed differentforces but found no significant differences (Table VI).DISCUSSION

    This review shows that miniscrews have been usedat success rates of 83% in orthodontic patients. By com-parison, palatal implants21 and miniplates3,16 reached90% to 95%. Dental implants have less than a 7% to9% risk of failure over 10 to 15 years.22,23 Miniscrewshave the advantage of simple surgical application, andan orthodontist can perform the procedure. Screwswith diameters of 1.2 mm or greater were universallyused at success rates above 70%.

    Another significant factor has been length. Chen etal10 increased the success rate from 72% to 90% by us-ing 8-mm instead of 6-mm long screws. Four studiesalso showed higher success rates with additional lengthat the same diameter but without a statistically signifi-

    ears 20-30 years .30 years Mean age

    3 88.2 85.0 21.8

    29.0

    100

    29.0

    3 100 17.9

    24.9

    15.5

    29.8

    29.9

    13.7

    5 82.4 100 21.8

    26.9

    3 91.9

    3* 84.1* 93.6* 24.5

    January 2010cant difference.Increasing screw diameter and length also raised the

    risk of root damage during placement.15 Miniscrews of1.2-mmdiameter and at least 8-mm length have sufficientstability with a minimum risk of root damage. Carefulplanning and radiographic evaluation of the placementsite can further minimize this risk.24,25 Although it hasbeen shown in animal studies that minimal root damageduring placement can be repaired on removal of thescrew,26 root contact or even proximity is also considereda major factor for the failure of miniscrews.27,28

    Considering the surgical protocols, a mucoperiostealflap is associated with more patient discomfort, swelling,and pain. The flapless method is less time-consumingbutmight hinder amore precise placement of the screw.12

    For both protocols, opposing success rates have beenpublished.3,12,13 Only 1 study reported higher successrates (100%) for screws placed with flap surgery.12

    However, only 10 screws were used with this procedure.Further investigations are needed to clarify this issue.

  • American Journal of Orthodontics and Dentofacial Orthopedics Crismani et al 111Table IV. Success rate (%) by screw length and diameter(mm)

    AuthorsScrews(n) Length Diameter

    Successrate

    Miyawaki et al3 10 6 1 0.0*

    101 11 1.5 83.9*

    23 14 2.3 85*

    Cheng et al4 48 5, 7 2 85.4

    31 9 2 93.5

    31 11 2 93.0

    20 13 2 85.0

    Volume 137, Number 1The angle between theminiscrews long axis and thecortical bone was evaluated only by Park et al.9 Withoutmajor differences in the success rates, it was argued thatplacing screws not perpendicular to the bone surface,but at an obtuse angle, lowered the risk of root damageand increased the screws contact with cortical bone.Liou et al29 analyzed the infrazygomatic crest in com-puted tomography scans and concluded that miniscrewsshould be placed at a steeper angle in this area.

    Screw success rates were higher in the maxilla thanin the mandible in all but 1 study.15 Three studies signif-icantly supported the maxilla as a more suitable place-ment site for miniscrews.4,9,16 All 3 studies interpreted

    10 15 2 90.0

    Liou et al5 32 17 2 100

    Fritz et al6 36 6, 8, 10 1.4, 1.6, 2 70.0

    Park et al7 (2005) 4 4 1.2 75.0

    14 6 1.2 86.7

    8 8 1.2 100

    2 10 1.2 100

    2 15 2 100

    Motoyoshi et al8 (2006) 124 8 1.6 85.5

    Park et al9 (2006) 19 5 1.2 84.2

    157 6-8 1.2 93.6

    46 4-10 1.2 89.1

    5 10-15 2 80.0

    Chen et al10 (2006) 18 6 1.2 72.2*

    41 8 1.2 90.2*

    Tseng et al11 15 8 2 80.0

    10 10 2 90.0

    12 12 2 100

    8 14 2 100

    Herman et al12 49 6-10 1.8 61.2

    Kuroda et al13 37 7, 11 2, 2.3 81.1

    13 6 1.3 69.2

    6 7 1.3 83.3

    45 8 1.3 93.3

    12 10 1.3 91.7

    3 12 1.3 100

    Wiechmann et al14 79 5-10 1.1 59.6*

    54 5-10 1.6 87.0*

    Motoyoshi et al15 (2007) 169 8 1.6 91.9

    Chen et al16 (2007) 72 4-10 1.2 76.4

    201 5-21 2 82.6

    *P\0.05.the lower success in the mandible as a consequence ofoverheating the bone during placement. Particular caremust therefore be used for pilot drilling, screw tighten-ing, and rinsing. In addition, mandibular miniscrewsmight be more exposed to masticatory interferences.

    Fig. Comparison of success rates for screws placedwith and without flap surgery.

    Table V. Success rate (%) by location of screw place-ment

    Maxilla Mandible

    AuthorsScrews(n)

    Successrate

    Screws(n)

    Successrate

    Miyawaki et al3 63 84.1 61 83.6

    Cheng et al4 93.3* 77.1*

    Liou et al5 32 100

    Fritz et al6 18 18

    Park et al7 (2005) 8 100 22 86.4

    Motoyoshi et al8 (2006) 80 88.8 44 79.5

    Park et al9 (2006) 124 96.0* 103 86.4*

    Chen et al10 (2006) 43 86.0 16 81.3

    Tseng et al11 27 96.3 18 83.3

    Herman et al12 49 61.2

    Kuroda et al13 61 91.8 18 77.8

    Wiechmann et al14 90 86.7 43 55.8

    Motoyoshi et al15 (2007) 100 84.0 69 87.0

    Chen et al16 (2007) 263 88.2* 96 77.1*

    Total 958 Mean: 89.0

    6 10.0

    508 Mean: 79.6

    6 8.7

    Weighted by number

    of screws

    Mean weighted

    by number

    of screws:

    87.9 6 7.6

    Mean weighted

    by number

    of screws:

    80.4 6 8.5

    *P\0.05.

  • tion

    ding

    12 89.8

    85.7

    112 Crismani et al American Journal of Orthodontics and Dentofacial OrthopedicsTable VI. Success rate (%) by latency period (days), dura

    Authors Screws (n) Time before loa

    Miyawaki et al3 20 \3029 30-90

    72 $90

    Cheng et al4 92 14-28

    Liou et al5 32 14

    Fritz et al6 36 \28Park et al7 (2005) 30 14

    Motoyoshi et al8 (2006) 124 Immediate

    Park et al9 (2006) 227

    Chen et al10 (2006) 59 14

    Tseng et al11 45 14

    Herman et al12 49 1-149

    Kuroda et al13 59 Immediate

    14 #30

    6 .30

    5

    55

    8

    11

    Wiechmann et al14 133 Immediate

    Motoyoshi et al15 (2007) 133 \3036 90

    Chen et al16 (2007) 98 7-28

    79 35-84

    40 $91Significantly higher success on the left side might bea statistical outlier. Unilateral preference of the mastica-tion side and potentially better oral hygiene on the leftside because of the prevalence of right-handed patientsmight be further explanations.

    A more critical issue is the determination ofa placement site. There are preferable areas that offersufficient bone and root distances. Poggio et al30 eval-uated tomographic images of mandibles and maxillaeto define safe zones for placing miniscrews. In themaxilla, they recommended interradicular spaces be-tween the canine and the second molar on the palatalside, and between the canine and the first molar onthe buccal side. In the mandible, they suggested inter-radicular spaces between the canine and the secondmolar.

    Details of screw design might play another role inscrew stability but were not evaluated in any of thesestudies. Comprehensive research on thread width andpitch or different head or end configurations has yet tobe published.

    Stability of dental implants relies on osseointegra-tion, which requires several weeks for the host bone toachieve intimate contact with the implant.31,32 Gener-ally, miniscrews are loaded sooner. No included studyrecommended longer healing periods for higher success83.3

    50 80.0

    100 89.1

    150 75.0

    200 100

    6 100-200 76.7

    6 200 82.0

    6 200 97.2

    100-300 85.7

    88.6

    92.5(months), and amount of loading (cN)

    Duration of loading Force Success rate

    12 \200 85.082.8

    87.5

    100-200 91.3

    9 400 100

    5.3 6 3.2 70.0

    12.3 6 5.7 \200 90.06 \200 85.515 \200 91.6 100-200 84.7

    100-200 91.1

    150 61.2

    January 2010rates. Therefore, miniscrews can be loaded within daysafter placement with forces up to 200 cN.

    Whereas miniscrews are sometimes believed toachieve their retention purely mechanically, the ideaof osseointegration should not be dismissed entirely.19

    Miniscrews with an acid-etched surface for enhancedosseointegration have been used successfully.33 Partialosseointegration has since been shown in both in-vitroand in-vivo studies.34,35

    CONCLUSIONS

    In all articles analyzed, the authors described thesuccess rates of miniscrews as sufficient for orthodontictreatment. This review further showed that screws of1.2-mm diameter and at least 8-mm length are prefera-ble, because they are stable and minimize the risk ofroot damage. The maxilla was shown to be better suitedfor miniscrews. As for the placement protocol, the datawere inconclusive for definite recommendations. Atcomparable success rates, the flapless method shouldbe chosen because it is less invasive and causes lesspatient discomfort.

    Immediate or early loading of miniscrews is possi-ble, since longer healing periods did not provide addi-tional stability at forces of up to 200 cN.

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    Miniscrews in orthodontic treatment: Review and analysis of published clinical trialsMaterial and methodsResultsDiscussionConclusionsReferences