Influence of Section Thickness on the Accuracy and Specificity of Histometric Parameters Using Confocal Laser Scanning Microscopy in a Canine Model of Experimental Peri-Implantitis—A Proof of Concept

Objectives: Tissue breakdown was assessed by confocal laser scanning microscopy (CLSM) using autofluorescence around implants with ligatures, on a dog hemimandible. Influence of section thickness on the accuracy of histometrical observations was also evaluated, in comparison with thin sections in light microscopy. Material and Methods: Three months after tooth extraction, implants were placed. Two months after abutment placement, ligatures were placed with no plaque control. 11 months post-implantation, the animal was sacrificed. Undecalcified thin (30 µm) sections were cut, stained and evaluated by light microscopy to be used as a reference. Additional sections were performed, so that another pair of unstained thick sections resulted (250–300 µm). Tissue loss was assessed using histomorphometric parameters under CLSM and was compared to the light microscopy reference ones. Results: Morphometry confirmed tissue loss more pronounced on the “thick” and quick sections, when compared to the time-consuming and technique-sensitive “thin” ones. Conclusions: Within the limits of the present study, the adequacy of histometrical observations under CLSM reveal commensurable information about soft-tissue-bone-implant details, when compared to traditional light microscopy histological protocols. The CLSM investigation may seem demanding, yet the richness of data acquired may justify this approach, provided seatbacks caused by improper manipulation of “thick” sections are avoided.

[1]  R. Drăgoi,et al.  Histomorphometrical and CBCT Evaluation of Tissue Loss Progression Induced by Consecutive, Alternate Ligatures in Experimental Peri-Implantitis in a Dog Model: A Pilot Study , 2022, Journal of clinical medicine.

[2]  Abdalla Abdal-hay,et al.  Fabrication of micropores on titanium implants using femtosecond laser technology: Perpendicular attachment of connective tissues as a pilot study , 2022, Optics & Laser Technology.

[3]  E. Maiorano,et al.  Peri‐implantitis‐like medication‐related osteonecrosis of the jaw: Clinical considerations and histological evaluation with confocal laser scanning microscope , 2021, Oral diseases.

[4]  Ulrich Dirnagl,et al.  The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research , 2020, British journal of pharmacology.

[5]  Ulrich Dirnagl,et al.  The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research* , 2020, BMC Veterinary Research.

[6]  C. Viegas,et al.  Evaluation of a novel dog animal model for peri-implant disease: clinical, radiographic, microbiological and histological assessment , 2020, Clinical Oral Investigations.

[7]  R. Weiss,et al.  Cone Beam Computed Tomography in Oral and Maxillofacial Surgery: An Evidence-Based Review , 2019, Dentistry journal.

[8]  Young-Jun Jeon,et al.  Osteogenic Cell Behavior on Titanium Surfaces in Hard Tissue , 2019, Journal of clinical medicine.

[9]  J. Derks,et al.  Peri‐implantitis , 2018, Journal of periodontology.

[10]  K. Wolff,et al.  Comparison of contact radiographed and stained histological sections for osseointegration analysis of dental implants: an in vivo study. , 2018, Oral surgery, oral medicine, oral pathology and oral radiology.

[11]  L. Capasso,et al.  Bone natural autofluorescence and confocal laser scanning microscopy: Preliminary results of a novel useful tool to distinguish between forensic and ancient human skeletal remains. , 2017, Forensic science international.

[12]  Y. Seol,et al.  An immediate peri‐implantitis induction model to study regenerative peri‐implantitis treatments , 2017, Clinical oral implants research.

[13]  Guang Hong,et al.  Biomechanical and histological evaluation of the osseointegration capacity of two types of zirconia implant , 2016, International journal of nanomedicine.

[14]  I. Abrahamsson,et al.  The effect of the local use of chlorhexidine in surgical treatment of experimental peri-implantitis in dogs. , 2015, Journal of clinical periodontology.

[15]  M. Dard,et al.  The Dog as a Model for Peri-Implantitis. A Review , 2014, Journal of investigative surgery : the official journal of the Academy of Surgical Research.

[16]  P. Coelho,et al.  Reimplantation of dental implants following ligature-induced peri-implantitis: a pilot study in dogs. , 2013, Clinical implant dentistry and related research.

[17]  I. Abrahamsson,et al.  Spontaneous progression of experimental peri-implantitis at implants with different surface characteristics: an experimental study in dogs. , 2012, Journal of clinical periodontology.

[18]  F. Moll,et al.  A Histological Analysis , 2012 .

[19]  K. Moharamzadeh,et al.  A review of histomorphometric analysis techniques for assessing implant-soft tissue interface , 2011, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[20]  T. Berglundh,et al.  Are peri-implantitis lesions different from periodontitis lesions? , 2011, Journal of clinical periodontology.

[21]  C. B. Johansson,et al.  3.313 – Histological Analysis , 2011 .

[22]  I. Abrahamsson,et al.  Implant surface characteristics influence the outcome of treatment of peri-implantitis: an experimental study in dogs. , 2011, Journal of clinical periodontology.

[23]  Stephan Saalfeld,et al.  Globally optimal stitching of tiled 3D microscopic image acquisitions , 2009, Bioinform..

[24]  N. Lang,et al.  Effects of decontamination and implant surface characteristics on re-osseointegration following treatment of peri-implantitis. , 2009, Clinical oral implants research.

[25]  I. Abrahamsson,et al.  Spontaneous progression of ligatured induced peri-implantitis at implants with different surface characteristics. An experimental study in dogs II: histological observations. , 2009, Clinical oral implants research.

[26]  Gwendolyn R. Goss,et al.  Theory and Practice of Histological Techniques , 2009 .

[27]  C. Johansson,et al.  Comparing light and fluorescence microscopic data : A pilot study of titanium and magnesium oxide implant integration in rabbit bone , 2009 .

[28]  K. Günther,et al.  Staining undecalcified bone sections a modified technique for an improved visualization of synthetic bone substitutes. , 2008, Acta chirurgiae orthopaedicae et traumatologiae Cechoslovaca.

[29]  I. Abrahamsson,et al.  Spontaneous progression of peri-implantitis at different types of implants. An experimental study in dogs. I: clinical and radiographic observations. , 2008, Clinical oral implants research.

[30]  L. Heitz‐Mayfield Peri-implant diseases: diagnosis and risk indicators. , 2008, Journal of clinical periodontology.

[31]  J. Wennström,et al.  Clinical characteristics at implants with a history of progressive bone loss. , 2008, Clinical oral implants research.

[32]  N. Lang,et al.  Spontaneous progression of ligature induced peri-implantitis at implants with different surface roughness: an experimental study in dogs. , 2007, Clinical oral implants research.

[33]  Haim Tal,et al.  Impact of implant overloading on the peri-implant bone in inflamed and non-inflamed peri-implant mucosa. , 2007, Clinical oral implants research.

[34]  M. Dard,et al.  Histological and immunohistochemical analysis of initial and early osseous integration at chemically modified and conventional SLA titanium implants: preliminary results of a pilot study in dogs. , 2007, Clinical oral implants research.

[35]  Stefan Renvert,et al.  Nine- to fourteen-year follow-up of implant treatment. Part II: presence of peri-implant lesions. , 2006, Journal of clinical periodontology.

[36]  U. Lekholm,et al.  Prevalence of subjects with progressive bone loss at implants. , 2005, Clinical oral implants research.

[37]  J. Lindhe,et al.  Spontaneous progression of experimentally induced periimplantitis. , 2004, Journal of clinical periodontology.

[38]  Yuehuei H. An,et al.  Handbook of Histology Methods for Bone and Cartilage , 2003, Humana Press.

[39]  Björn Klinge,et al.  A systematic review of the incidence of biological and technical complications in implant dentistry reported in prospective longitudinal studies of at least 5 years. , 2002, Journal of clinical periodontology.

[40]  K Gotfredsen,et al.  Bone reactions at implants subjected to experimental peri-implantitis and static load. A study in the dog. , 2002, Journal of clinical periodontology.

[41]  D. Cochran,et al.  Biologic Width around one- and two-piece titanium implants. , 2001, Clinical oral implants research.

[42]  M. Spector,et al.  Connective tissue orientation around dental implants in a canine model. , 2001, Clinical oral implants research.

[43]  F. Nociti,et al.  Absorbable versus nonabsorbable membranes and bone grafts in the treatment of ligature-induced peri-implantitis defects in dogs: a histometric investigation. , 2001, The International journal of oral & maxillofacial implants.

[44]  A. Hefti,et al.  Confocal laser scanning microscopy and scanning electron microscopy of tissue Ti-implant interfaces. , 2001, Micron.

[45]  R. Haas,et al.  Experimentally induced peri-implantitis: a review of different treatment methods described in the literature. , 2000, The International journal of oral & maxillofacial implants.

[46]  D Buser,et al.  Biologic width around titanium implants. A physiologically formed and stable dimension over time. , 2000, Clinical oral implants research.

[47]  N. Lang,et al.  Attempts to obtain re-osseointegration following experimental peri-implantitis in dogs. , 1999, Clinical oral implants research.

[48]  M. Hürzeler,et al.  Treatment of peri-implantitis using guided bone regeneration and bone grafts, alone or in combination, in beagle dogs. Part 2: Histologic findings. , 1997, The International journal of oral & maxillofacial implants.

[49]  Y. Ayukawa,et al.  Study of bone formation around dense hydroxyapatite implants using light microscopy, image processing and confocal laser scanning microscopy. , 1997, Biomaterials.

[50]  J. Fiorellini,et al.  Comparison of healed tissues adjacent to submerged and non-submerged unloaded titanium dental implants. A histometric study in beagle dogs. , 1996, Clinical oral implants research.

[51]  P. Glantz,et al.  Resolution of ligature-induced peri-implantitis lesions in the dog. , 1995, Journal of clinical periodontology.

[52]  P Morberg,et al.  Importance of ground section thickness for reliable histomorphometrical results. , 1995, Biomaterials.

[53]  P. Trisi,et al.  Histochemical and confocal laser scanning microscopy study of the bone-titanium interface: an experimental study in rabbits. , 1994, Biomaterials.

[54]  J. Strub,et al.  Treatment of ligature-induced peri-implantitis using guided tissue regeneration: a clinical and histologic study in the beagle dog. , 1993, The International journal of oral & maxillofacial implants.

[55]  J. Fiorellini,et al.  Soft tissue reactions to non-submerged unloaded titanium implants in beagle dogs. , 1992, Journal of periodontology.

[56]  J. Lindhe,et al.  Experimental breakdown of peri-implant and periodontal tissues. A study in the beagle dog. , 1992, Clinical oral implants research.

[57]  A. Malcolm,et al.  Control of rapid nitric acid decalcification. , 1984, Journal of clinical pathology.

[58]  K. Donath,et al.  A method for the study of undecalcified bones and teeth with attached soft tissues. The Säge-Schliff (sawing and grinding) technique. , 1982, Journal of oral pathology.

[59]  J. Eitenmüller,et al.  Preparation of histological sections from nondecalcified cortical bone. , 1976, Zeitschrift fur mikroskopisch-anatomische Forschung.

[60]  J. Lindhe,et al.  Experimental periodontal breakdown in the dog. , 1975, Scandinavian journal of dental research.

[61]  M. Kalweit The preparation of thin sections of undecalcified bone and surrounding soft tissues. , 1971, Canadian journal of medical technology.

[62]  R. Ziegler,et al.  [A simple methylmetacrylate embedding for the preparation of thin sections from bones]. , 1970, Zeitschrift fur wissenschaftliche Mikroskopie und mikroskopische Technik.

[63]  H. Frost,et al.  Preparation of thin undecalcified bone sections by rapid manual method. , 1958, Stain technology.