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Can bone marrow aspirate concentrate change the mineralization pattern of the anterior maxilla treated with xenografts? A preliminary study

Identifieur interne : 000616 ( Pmc/Corpus ); précédent : 000615; suivant : 000617

Can bone marrow aspirate concentrate change the mineralization pattern of the anterior maxilla treated with xenografts? A preliminary study

Auteurs : André Antonio Pelegrine ; Marcelo Lucchesi Teixeira ; Marcelo Sperandio ; Thiago Sousa Almada ; Karl Erik Kahnberg ; Paulo José Pasquali ; Antonio Carlos Aloise

Source :

RBID : PMC:4792049

Abstract

Objective:

To evaluate bony reconstruction of the atrophic anterior maxilla using particulate grafts with or without autologous bone marrow aspirate concentrate (BMAC).

Materials and Methods:

Eight patients with atrophy of the anterior maxilla due to teeth loss were selected and split into groups according to the type of material used: Control Group (CG) (n = 4) - particulate xenograft only and Test Group (TG) (n = 4) - a combination of particulate xenograft and BMAC. Both groups received a collagen membrane to cover the xenograft. After 4 months, during implant placement, a sample of bone was removed from the graft area using a 2 mm diameter trephine bur. The specimens were fixed and preserved for histomorphometric evaluation, which included the following parameters: Mineralized tissue (MT) and non-MT (NMT). Cone beam computed tomography was performed at 3 time intervals to measure bone thickness: (1) Before grafting, (2) 4 months and (3) 8 months postgrafting, using localized bone gain (mm) as the outcome variable.

Results:

Tomographic analysis revealed bone gain in CG of 3.78 ± 1.35 mm and 4.34 ± 1.58 mm at 4 and 8 months, respectively. TG showed an increase of 3.79 ± 0.52 mm and 4.09 ± 1.33 mm after 4 and 8 months, respectively. Histomorphometric analysis revealed that, for CG, MT- and NMT-related values were 52.3% ± 16.78% and 47.70% ± 5.55%, respectively, whereas for TG, they were 65.04% ± 20.98% and 34.96 ± 10.38, respectively.

Conclusion:

Although radiographic bone gain appeared similar between the groups, the use of BMAC obtained via the BMAC® method revealed an increased mineralization trend in the anterior maxilla. It must be highlighted, however, that this is a preliminary study with a relatively small sample population and further studies with larger sample sizes are needed to verify these results.


Url:
DOI: 10.4103/0976-237X.177112
PubMed: 27041895
PubMed Central: 4792049

Links to Exploration step

PMC:4792049

Le document en format XML

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<p>To evaluate bony reconstruction of the atrophic anterior maxilla using particulate grafts with or without autologous bone marrow aspirate concentrate (BMAC).</p>
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<sec id="st2">
<title>Materials and Methods:</title>
<p>Eight patients with atrophy of the anterior maxilla due to teeth loss were selected and split into groups according to the type of material used: Control Group (CG) (
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= 4) - particulate xenograft only and Test Group (TG) (
<italic>n</italic>
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<title>Results:</title>
<p>Tomographic analysis revealed bone gain in CG of 3.78 ± 1.35 mm and 4.34 ± 1.58 mm at 4 and 8 months, respectively. TG showed an increase of 3.79 ± 0.52 mm and 4.09 ± 1.33 mm after 4 and 8 months, respectively. Histomorphometric analysis revealed that, for CG, MT- and NMT-related values were 52.3% ± 16.78% and 47.70% ± 5.55%, respectively, whereas for TG, they were 65.04% ± 20.98% and 34.96 ± 10.38, respectively.</p>
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<journal-id journal-id-type="nlm-ta">Contemp Clin Dent</journal-id>
<journal-id journal-id-type="iso-abbrev">Contemp Clin Dent</journal-id>
<journal-id journal-id-type="publisher-id">CCD</journal-id>
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<issn pub-type="epub">0976-2361</issn>
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<article-id pub-id-type="pmc">4792049</article-id>
<article-id pub-id-type="publisher-id">CCD-7-21</article-id>
<article-id pub-id-type="doi">10.4103/0976-237X.177112</article-id>
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<subject>Original Article</subject>
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<article-title>Can bone marrow aspirate concentrate change the mineralization pattern of the anterior maxilla treated with xenografts? A preliminary study</article-title>
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<name>
<surname>Pelegrine</surname>
<given-names>André Antonio</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
<xref ref-type="corresp" rid="cor1"></xref>
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<name>
<surname>Teixeira</surname>
<given-names>Marcelo Lucchesi</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
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<name>
<surname>Sperandio</surname>
<given-names>Marcelo</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Almada</surname>
<given-names>Thiago Sousa</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Kahnberg</surname>
<given-names>Karl Erik</given-names>
</name>
<xref ref-type="aff" rid="aff2">1</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Pasquali</surname>
<given-names>Paulo José</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Aloise</surname>
<given-names>Antonio Carlos</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
</contrib-group>
<aff id="aff1">
<italic>Department of Implantology, São Leopoldo Mandic Institute and Research Center, Campinas, Brazil</italic>
</aff>
<aff id="aff2">
<label>1</label>
<italic>Department of Oral Surgery, University of Gothenburg, Gothenburg, Sweden</italic>
</aff>
<author-notes>
<corresp id="cor1">
<bold>Correspondence:</bold>
Prof. André Antonio Pelegrine, Rua das Areias, 37, CEP 13024-530, São Paulo, Brazil. E-mail:
<email xlink:href="pelegrineandre@gmail.com">pelegrineandre@gmail.com</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub">
<season>Jan-Mar</season>
<year>2016</year>
</pub-date>
<volume>7</volume>
<issue>1</issue>
<fpage>21</fpage>
<lpage>26</lpage>
<permissions>
<copyright-statement>Copyright: © Contemporary Clinical Dentistry</copyright-statement>
<copyright-year>2016</copyright-year>
<license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc-sa/3.0">
<license-p>This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.</license-p>
</license>
</permissions>
<abstract>
<sec id="st1">
<title>Objective:</title>
<p>To evaluate bony reconstruction of the atrophic anterior maxilla using particulate grafts with or without autologous bone marrow aspirate concentrate (BMAC).</p>
</sec>
<sec id="st2">
<title>Materials and Methods:</title>
<p>Eight patients with atrophy of the anterior maxilla due to teeth loss were selected and split into groups according to the type of material used: Control Group (CG) (
<italic>n</italic>
= 4) - particulate xenograft only and Test Group (TG) (
<italic>n</italic>
= 4) - a combination of particulate xenograft and BMAC. Both groups received a collagen membrane to cover the xenograft. After 4 months, during implant placement, a sample of bone was removed from the graft area using a 2 mm diameter trephine bur. The specimens were fixed and preserved for histomorphometric evaluation, which included the following parameters: Mineralized tissue (MT) and non-MT (NMT). Cone beam computed tomography was performed at 3 time intervals to measure bone thickness: (1) Before grafting, (2) 4 months and (3) 8 months postgrafting, using localized bone gain (mm) as the outcome variable.</p>
</sec>
<sec id="st3">
<title>Results:</title>
<p>Tomographic analysis revealed bone gain in CG of 3.78 ± 1.35 mm and 4.34 ± 1.58 mm at 4 and 8 months, respectively. TG showed an increase of 3.79 ± 0.52 mm and 4.09 ± 1.33 mm after 4 and 8 months, respectively. Histomorphometric analysis revealed that, for CG, MT- and NMT-related values were 52.3% ± 16.78% and 47.70% ± 5.55%, respectively, whereas for TG, they were 65.04% ± 20.98% and 34.96 ± 10.38, respectively.</p>
</sec>
<sec id="st4">
<title>Conclusion:</title>
<p>Although radiographic bone gain appeared similar between the groups, the use of BMAC obtained via the BMAC
<sup>®</sup>
method revealed an increased mineralization trend in the anterior maxilla. It must be highlighted, however, that this is a preliminary study with a relatively small sample population and further studies with larger sample sizes are needed to verify these results.</p>
</sec>
</abstract>
<kwd-group>
<kwd>Bone marrow</kwd>
<kwd>bone regeneration</kwd>
<kwd>stem cells</kwd>
</kwd-group>
</article-meta>
</front>
<body>
<sec sec-type="intro" id="sec1-1">
<title>Introduction</title>
<p>Bone loss in the edentulous anterior maxilla is a common occurrence, with a 35% bone resorption rate expected within 6 months of teeth extraction.[
<xref rid="ref1" ref-type="bibr">1</xref>
] Loss of bone thickness precedes loss of height, which explains why appositional approaches aimed at thickness augmentation are the most prevalent.[
<xref rid="ref2" ref-type="bibr">2</xref>
] In appositional bone reconstructions, autologous bone grafts would be considered the gold standard.[
<xref rid="ref3" ref-type="bibr">3</xref>
] The morbidity associated with autologous bone grafting is, however, the main limitation of this technique for both the professionals, due to technical difficulties, and the patients, who remain reluctant to undergo more invasive surgical procedures.[
<xref rid="ref2" ref-type="bibr">2</xref>
<xref rid="ref3" ref-type="bibr">3</xref>
<xref rid="ref4" ref-type="bibr">4</xref>
] On the other hand, the use of bone marrow as the source of cells in bony tissue engineering represents an alternative to traditional autologous bone grafting, since, in addition to showing good regenerative results, the risk of complications related to aspiration is 10 times lower than observed with the classic iliac crest grafts. At last, when complications do occur using the former approach, they are much less severe than the latter.[
<xref rid="ref5" ref-type="bibr">5</xref>
]</p>
<p>There are only very few studies in the literature reporting anterior maxillary reconstructions using particulate bone grafts, which hinders decision making by the surgeon in terms of the type of graft to use. Xenografts, which are available in a particulate form, are becoming a more viable option than autologous grafts, despite their inability to promote osteogenesis and osteoinduction, a frequent criticism regarding this approach.[
<xref rid="ref6" ref-type="bibr">6</xref>
]</p>
<p>A series of studies in experimental models attempted to associate particulate xenografts with cell therapy, aiming to achieve osteogenesis and osteoinduction. The authors concluded that the bone marrow was the ideal source of cells, particularly in its concentrated form.[
<xref rid="ref6" ref-type="bibr">6</xref>
<xref rid="ref7" ref-type="bibr">7</xref>
]</p>
<p>Therefore, the aim of the current study was to evaluate human bone reconstruction in the atrophic anterior maxilla, using particulate grafts with or without autologous bone marrow aspirate concentrate (BMAC).</p>
</sec>
<sec sec-type="materials|methods" id="sec1-2">
<title>Materials and Methods</title>
<p>This study was performed in the outpatient clinic of the Department of Implant Dentistry, São Leopoldo Mandic Dental School (Campinas, SP, Brazil), following approval by the research ethics committee (838.635/2014). Free and informed consent was obtained from all patients.</p>
<p>The inclusion criteria were patients lacking the four upper incisors, yet presenting both canines, who presented with no more than 3 mm of remaining alveolar ridge and needed anterior, maxillary implants. The patients committed to attending follow-up appointments and maintaining adequate oral hygiene. Patients with a history of neoplastic disease treated with radio or chemotherapy, pregnant or breastfeeding women, systemic diseases or treatments affecting bone homeostasis, allergy to components of the materials used, sinus pathologies, and smokers were excluded from the study.</p>
<p>Alveolar ridge thickness was measured using cone-beam computed tomography (CBCT, i-CAT Classic, Imaging Sciences International, Hatfield, PA, USA) in cross-sectional cuts of the generated images (Digital Imaging and Communications in Medicine-based data sets) at a resolution of 96 dpi, 14 bits grayscale, and 0.25 mm voxel size. The computed tomography (CT) unit was set at 120 kVp, 5 mA, and 20 s exposure.</p>
<p>Eight patients with a mean age of 52.4 ± 2.2 years were included in this study. All patients presented with an atrophic anterior maxilla for grafting before implant placement. The patients were randomly distributed using a web-based software available at
<ext-link ext-link-type="uri" xlink:href="www.randomization.com">www.randomization.com</ext-link>
into two groups according to material used: Control Group (CG) (
<italic>n</italic>
= 4) particulate bone xenograft only (Bio-Gen granules 500–1000 μm Bioteck, Vicenza, Italy), and Test Group (TG) (
<italic>n</italic>
= 4) with particulate bone xenograft combined with bone marrow concentrate obtained via the BMAC method. Following the principles of guided bone regeneration (GBR), collagen membranes (Biocollagen Bioteck, Vicenza, Italy) were placed over the bone grafts in all maxillary augmentation procedures in both groups.</p>
<p>At the end of the study, all patients were dentally rehabilitated using osseointegrated implants and fixed prostheses.</p>
<sec id="sec2-1">
<title>Bone marrow aspirate concentrate method</title>
<p>According to the manufacturer's instructions, bone marrow was harvested and processed in the operating room using the BMAC system (Bone Marrow Procedure Pack; Harvest Technologies, Plymouth, MA, USA). Briefly, in an outpatient setting and using local anesthesia (2% xylocaine without a vasoconstrictor), 30 mL of bone marrow aspirate was obtained from all patients via a puncture 2 cm laterocaudally from the upper posterior iliac crest, using a bone marrow needle (included in the pack) and heparinized 30 mL syringes (1 mL of 5.000 U/mL heparin).</p>
<p>The 30 mL bone marrow-filled syringe was connected to a filter bag, to which 8 mL of Anticoagulant Citrate Dextrose (ACD-A) anticoagulant was added. Following homogenization, a new syringe was attached and the filtered 30 mL removed. The bone marrow aspirate was then transferred into specific process disposables, which were placed in a SmartPReP2 centrifuge. After 14 min of centrifugation, two phases were obtained within the tube, i.e., the plasma supernatant and the precipitated bone marrow cell concentrate [Figure
<xref ref-type="fig" rid="F1">1a</xref>
and
<xref ref-type="fig" rid="F1">b</xref>
]. The plasma was removed using specific syringes provided in the kit; the cell concentrate was suspended and approximately 4 mL aspirated.</p>
<fig id="F1" position="float">
<label>Figure 1</label>
<caption>
<p>(a) Bone marrow after centrifugation (note the plasma supernatant being discarded); (b) aspiration of concentrated bone marrow cells</p>
</caption>
<graphic xlink:href="CCD-7-21-g001"></graphic>
</fig>
</sec>
<sec id="sec2-2">
<title>Surgical procedure</title>
<p>All patients were treated under local anesthesia (Mepiadre 2%, DFL, São Paulo, Brazil), and a full thickness flap was raised to provide access to the resorbed alveolar ridge. A carbide burr (Ar N 701 21 mm Jota Rotatory Instruments, Ruthi, Switzerland) was used for decortication with the aim of enhancing vascularization [
<xref ref-type="fig" rid="F2">Figure 2</xref>
].</p>
<fig id="F2" position="float">
<label>Figure 2</label>
<caption>
<p>Surgical site following flap elevation and decortication</p>
</caption>
<graphic xlink:href="CCD-7-21-g002"></graphic>
</fig>
<p>In both groups, the particulate bone graft was spread over the bone to cover the entire exposed area evenly, to achieve sufficient thickness [
<xref ref-type="fig" rid="F3">Figure 3</xref>
].</p>
<fig id="F3" position="float">
<label>Figure 3</label>
<caption>
<p>Graft in position</p>
</caption>
<graphic xlink:href="CCD-7-21-g003"></graphic>
</fig>
<p>In TG, the bone graft was mixed with bone marrow before placement at the site of the defect [
<xref ref-type="fig" rid="F4">Figure 4</xref>
]. Both groups were covered with an equine collagen membrane. The flaps were repositioned to completely cover the grafts and subsequently sutured with interrupted single 4–0 nylon sutures.</p>
<fig id="F4" position="float">
<label>Figure 4</label>
<caption>
<p>Xenograft combined with bone marrow aspirate concentrate</p>
</caption>
<graphic xlink:href="CCD-7-21-g004"></graphic>
</fig>
</sec>
<sec id="sec2-3">
<title>Computed tomography analysis</title>
<p>All patients were scanned at three different intervals:</p>
<p>(1) Baseline, or immediately before grafting; (2) 4 months after grafting; and (3) 8 months after the grafting procedure. For every CT slice, one tagged image file (TIF) image was generated. Assessment of the TIF images was performed using dedicated software (ImageJ; NIH, Bethesda, MD, USA) [Figure
<xref ref-type="fig" rid="F5">5a</xref>
-
<xref ref-type="fig" rid="F5">c</xref>
].</p>
<fig id="F5" position="float">
<label>Figure 5</label>
<caption>
<p>(a) Computed tomography image before treatment; (b) computed tomography image 4 months after treatment; (c) computed tomography image 8 months after treatment</p>
</caption>
<graphic xlink:href="CCD-7-21-g005"></graphic>
</fig>
<p>Thickness was measured in three points (apical, middle, and coronal), and an average was made per site. All measurements were performed twice by a single experienced blinded examiner andré antonio pelegrine (AAP), with a 15-day interval in between.</p>
</sec>
<sec id="sec2-4">
<title>Bone biopsies: Clinical procedure and evaluation</title>
<p>Dental implant placement was performed 4 months after grafting [Figure
<xref ref-type="fig" rid="F6">6a</xref>
and
<xref ref-type="fig" rid="F6">b</xref>
].</p>
<fig id="F6" position="float">
<label>Figure 6</label>
<caption>
<p>(a) Grafted site 4 months after reopening; (b) transoperative view, immediately after implant placement</p>
</caption>
<graphic xlink:href="CCD-7-21-g006"></graphic>
</fig>
<p>During the procedure, a cylindrical biopsy was retrieved from the implant placement site of each patient, using a trephine bur (2 mm internal Ø). The biopsies were routinely fixed, decalcified, dehydrated, embedded in paraffin, and sectioned. Four 7 μm sections were stained using Masson's Trichrome for histological and histomorphometrical analysis, representing the central aspect of each cylindrical biopsy. The relative quantity (%) of mineralized tissue (MT) and non-MT (NMT) were measured using the ImageJ software (NIH, Bethesda, MD, USA) [Figures
<xref ref-type="fig" rid="F7">7a</xref>
and
<xref ref-type="fig" rid="F7">b</xref>
]. All histomorphometric analyses were performed twice by a single experienced blinded examiner antonio carlos aloise (ACA), with a 15-day interval in between.</p>
<fig id="F7" position="float">
<label>Figure 7</label>
<caption>
<p>(a) Photomicrograph of a histological section stained with Masson's Trichrome, from the Control Group (×100); (b) photomicrograph of a histological slide stained with Masson's Trichrome from the Test Group (×100)</p>
</caption>
<graphic xlink:href="CCD-7-21-g007"></graphic>
</fig>
</sec>
<sec id="sec2-5">
<title>Statistical analysis</title>
<p>Commercially available software (GraphPad Prism 6.0 for Windows, GraphPad Software Inc., La Jolla, CA, USA) was used to compare all assessed parameters and to create charts. Due to the small sample size, data were considered not conforming to a normal distribution. The Wilcoxon test was therefore used, at a significance level of
<italic>P</italic>
< 0.05.</p>
</sec>
</sec>
<sec sec-type="results" id="sec1-3">
<title>Results</title>
<p>A minimum of two implants was placed in each previously grafted sites, all of which were osseointegrated and loaded after a 4-month healing period. At that point, the CT showed a gain in bone thickness of 3.78 ± 1.35 mm and 3.79 ± 0.52 mm for the CG and TG, respectively. After 8 months, the same areas recorded 4.34 ± 1.58 mm and 4.09 ± 1.33 mm for CG and TG, respectively [
<xref ref-type="table" rid="T1">Table 1</xref>
].</p>
<table-wrap id="T1" position="float">
<label>Table 1</label>
<caption>
<p>Bone thickness gain measured by computed tomography (in mm)</p>
</caption>
<graphic xlink:href="CCD-7-21-g008"></graphic>
</table-wrap>
<p>Regarding the histomorphometric analysis, CG showed values for MT and NMT of 52.3% ±16.78% and 47.70% ± 5.55%, respectively. TG showed values for MT and NMT of 65.04% ± 20.98% and 34.961 ± 10.38, respectively [
<xref ref-type="table" rid="T2">Table 2</xref>
].</p>
<table-wrap id="T2" position="float">
<label>Table 2</label>
<caption>
<p>Histomorphometric analysis (%)</p>
</caption>
<graphic xlink:href="CCD-7-21-g009"></graphic>
</table-wrap>
</sec>
<sec sec-type="discussion" id="sec1-4">
<title>Discussion</title>
<p>Alveolar bone atrophy is a common finding in the edentulous anterior maxilla,[
<xref rid="ref1" ref-type="bibr">1</xref>
<xref rid="ref8" ref-type="bibr">8</xref>
] consequently generating a high demand for bone reconstruction in this area prior to osseointegrated implant placement.[
<xref rid="ref9" ref-type="bibr">9</xref>
]</p>
<p>In the present study, all patients had an anterior maxillary bone thickness of <3 mm and both groups presented a thickness gain of approximately 4 mm, which was confirmed using CT. This level of bone gain is comparable to other studies that focused on reconstruction of the anterior maxilla.[
<xref rid="ref10" ref-type="bibr">10</xref>
<xref rid="ref11" ref-type="bibr">11</xref>
]</p>
<p>Some studies have evaluated the potential for bone regeneration using a combination of bone substitutes with GBR techniques. The latter technique was used in the present study since over the years, the use of membranes or tissue barriers has been shown to prevent the transfer of unwanted cells derived from adjacent soft tissues during the healing process.[
<xref rid="ref12" ref-type="bibr">12</xref>
<xref rid="ref13" ref-type="bibr">13</xref>
<xref rid="ref14" ref-type="bibr">14</xref>
]</p>
<p>CT analysis, both before (4 months after grafting) and after implant placement (8 months following grafting), revealed no volumetric loss between these time intervals (
<italic>P</italic>
> 0.05). There was, in fact, a numeric gain between 4 and 8 months, which most likely occurred secondary to the expanding characteristic of the macro-engineering of the implants used.</p>
<p>Regarding the histomorphometric findings, no significant difference was observed in either NMT or MT levels between the groups (
<italic>P</italic>
> 0.05);, however, a trend was observed for higher MT and lower NMT levels in TG. This could be justified by the presence of potentially osteogenic cells and growth factors with osteoinducing potential in the bone marrow, as reported by Pelegrine
<italic>et al</italic>
.[
<xref rid="ref6" ref-type="bibr">6</xref>
] and Aloise
<italic>et al</italic>
.[
<xref rid="ref15" ref-type="bibr">15</xref>
] According to Hermann
<italic>et al</italic>
.,[
<xref rid="ref16" ref-type="bibr">16</xref>
] the use of the closed BMAC system revealed findings similar to, or even better than those of more elaborate methods such as the isolation of mononuclear cells from the bone marrow using the FICOLL method. Sauerbier
<italic>et al</italic>
.[
<xref rid="ref17" ref-type="bibr">17</xref>
] stated that the closed BMAC system was an appropriate substitute for the open FICOLL system for bone regeneration procedures. Rickert
<italic>et al</italic>
. (2011)[
<xref rid="ref18" ref-type="bibr">18</xref>
] reported that the BMAC system could be used as an alternative to autologous bone grafting, therefore justifying its use in daily clinical practice. However, despite the fact of the low complications rate regarding bone marrow harvest, especially when compared to the autogenous bone graft harvest,[
<xref rid="ref5" ref-type="bibr">5</xref>
] it is important to consider that the use a xenograft without mixing it with the BMAC has, obviously, no complication related to a donor site. Therefore, the use of BMAC system associated to a xenograft has the advantage of achieving a higher mineralization pattern but still demands an invasive tissue harvest that may have repercussion in a routine clinical use.</p>
<p>In the present study, only the total MT was considered in detriment of the amount of vital and nonvital mineralized tissue. This decision was made based on the difficulty to distinguish between them in photomicrographs [Figure
<xref ref-type="fig" rid="F7">7a</xref>
and
<xref ref-type="fig" rid="F7">b</xref>
]. Two factors may have contributed to this:</p>
<p>(1) The staining method (Masson's Trichrome) not allowing such distinction based solely on difference in color, thus confusing the newly formed bone with residual graft particles; (2) the newly formed bone, which is normally disorganized and contains nucleated cell, and the residual graft particles, which show a lamellar pattern and do not contain any cell nuclei, did not match the histological images. The photomicrographs revealed images compatible with lamellar bone containing nucleated cells, which could relate to the presence of native bone within the trephine biopsy whereas the disorganized bone with no cell nuclei could be an artifact from tissue preparation. Such features hinder accurate segregation between vital and nonvital mineralized tissue.</p>
<p>Despite a trend showing higher MT when using the BMAC system, one must highlight that this is a preliminary study with a small study population and that there are not several studies investigating biomaterials combined with the techniques described. Despite the fact that further larger studies are needed to validate such findings, it is important to state that the safety and efficacy of the procedure are proven and established in maxillofacial surgery, especially in areas of residual alveolar ridge resorption prior to implacement.[
<xref rid="ref19" ref-type="bibr">19</xref>
]</p>
</sec>
<sec sec-type="conclusion" id="sec1-5">
<title>Conclusion</title>
<p>Although radiographic bone gain appeared similar between the groups, the use of concentrated bone marrow aspirate obtained via the BMAC system revealed a higher MT trend in maxillary reconstructions. It must be highlighted, however, that this is a preliminary study based on a small sample population and larger studies are needed to verify these results.</p>
<sec id="sec2-6">
<title>Financial support and sponsorship</title>
<p>Nil.</p>
</sec>
<sec id="sec2-7">
<title>Conflicts of interest</title>
<p>There are no conflicts of interest.</p>
</sec>
</sec>
</body>
<back>
<ack>
<title>Acknowledgment</title>
<p>The authors wish to thank Dr. Adriana Zardo João, for harvesting the bone marrow used in this study.</p>
</ack>
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