Name: surgical bone implantation technique for rat tibia models of diabetes and osteoporosis
Uploaded: 2024-07-05T12:40:00
Description: The rat has long served as a valuable animal model in implant dentistry and orthopedics, particularly in studying the interactions between biomaterials ...

The authors thank the Spanish State Research Agency for financial support through projects PID2020-114019RBI00 and PID2021-125150OB-I00.

All experimental procedures were conducted in accordance with the European Community Guidelines for the protection of animals used for scientific purposes (Directive 2010/63/EU) as implemented in Spanish law (Royal Decree 53/2013) and Generalitat de Catalunya regulations (Decree 214/97). Ethics approval for all animal procedures and handling was obtained from the Ethics Committee for Animal Experimentation of the Vall D&#39;Hebron Institut de Recerca (registration number 72/18 CEEA). For the osteoporotic model, female Sp...

<ol>
	<li>Turner, R. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+osteoporosis.">Animal models for osteoporosis.</a> Rev Endocr Metab Disord. 2 (1), 117-127 (2001).</li><li>Jee, W. S., Yao, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overview:+Animal+models+of+osteopenia+and+osteoporosis.">Overview: Animal models of osteopenia and osteoporosis.</a> J Musculoskelet Neuronal Interact. 1 (3), 193-207 (2001).</li><li>Kim, B. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+structural+and+hormonal+basis+of+sex+differences+in+peak+appendicular+bone+strength+in+rats.">The structural and hormonal basis of sex differences in peak appendicular bone strength in rats.</a> J Bone Miner Res. 18 (1), 150-155 (2003).</li><li>Alenezi, A., Galli, S., Atefyekta, S., Andersson, M., Wennerberg, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osseointegration+effects+of+local+release+of+strontium+ranelate+from+implant+surfaces+in+rats.">Osseointegration effects of local release of strontium ranelate from implant surfaces in rats.</a> J Mater Sci Mater Med. 30 (10), 116 (2019).</li><li>Blanc-Sylvestre, N., Bouchard, P., Chaussain, C., Bardet, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-clinical+models+in+implant+dentistry:+Past,+present,+future.">Pre-clinical models in implant dentistry: Past, present, future.</a> Biomedicines. 9 (11), 1538 (2021).</li><li>Schliephake, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functionalization+of+titanium+implants+using+a+modular+system+for+binding+and+release+of+VEGF+enhances+bone-implant+contact+in+a+rodent+model.">Functionalization of titanium implants using a modular system for binding and release of VEGF enhances bone-implant contact in a rodent model.</a> J Clin Periodontol. 42 (3), 302-310 (2015).</li><li>Mafra, C. E. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+different+doses+of+synthetic+parathyroid+hormone+(1-34)+on+bone+around+implants:+A+preclinical+rat+model.">Effect of different doses of synthetic parathyroid hormone (1-34) on bone around implants: A preclinical rat model.</a> Braz Dent J. 30 (1), 43-46 (2019).</li><li>Rybaczek, T., Tangl, S., Dobsak, T., Gruber, R., Kuchler, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+parathyroid+hormone+on+osseointegration+in+insulin-treated+diabetic+rats.">The effect of parathyroid hormone on osseointegration in insulin-treated diabetic rats.</a> Implant Dent. 24 (4), 392-396 (2015).</li><li>Zou, G. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+local+delivery+of+bfgf+from+plga+microspheres+on+osseointegration+around+implants+in+diabetic+rats.">Effects of local delivery of bfgf from plga microspheres on osseointegration around implants in diabetic rats.</a> Oral Surg Oral Med Oral Pathol Oral Radiol. 114 (3), 284-289 (2012).</li><li>Zhang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+nerve+growth+factor+on+osseointegration+of+titanium+implants+in+type+2+diabetic+rats.">Effect of nerve growth factor on osseointegration of titanium implants in type 2 diabetic rats.</a> Int J Oral Maxillofac Implants. 31 (5), 1189-1194 (2016).</li><li>Kuchler, U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intermittent+parathyroid+hormone+fails+to+stimulate+osseointegration+in+diabetic+rats.">Intermittent parathyroid hormone fails to stimulate osseointegration in diabetic rats.</a> Clin Oral Implants Res. 22 (5), 518-523 (2011).</li><li>Hashiguchi, C., Kawamoto, S., Kasai, T., Nishi, Y., Nagaoka, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+an+antidiabetic+drug+on+biomechanical+and+histological+parameters+around+implants+in+type+2+diabetic+rats.">Influence of an antidiabetic drug on biomechanical and histological parameters around implants in type 2 diabetic rats.</a> Implant Dent. 23 (3), 264-269 (2014).</li><li>Han, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+topical+delivery+of+insulin+from+fibrin+gel+loaded+with+poly(lactic-co-glycolic+acid)+microspheres+improves+the+biomechanical+retention+of+titanium+implants+in+type+1+diabetic+rats.">Sustained topical delivery of insulin from fibrin gel loaded with poly(lactic-co-glycolic acid) microspheres improves the biomechanical retention of titanium implants in type 1 diabetic rats.</a> J Oral Maxillofac Surg. 70 (10), 2299-2308 (2012).</li><li>De Molon, R. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+diabetes+mellitus+and+metabolic+control+on+bone+healing+around+osseointegrated+implants:+Removal+torque+and+histomorphometric+analysis+in+rats.">Impact of diabetes mellitus and metabolic control on bone healing around osseointegrated implants: Removal torque and histomorphometric analysis in rats.</a> Clin Oral Implants Res. 24 (7), 831-837 (2013).</li><li>Simon, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+phenotypic+and+genomic+analysis+of+c57bl/6j+and+c57bl/6n+mouse+strains.">A comparative phenotypic and genomic analysis of c57bl/6j and c57bl/6n mouse strains.</a> Genome Biol. 14 (7), R82 (2013).</li><li>Cirano, F. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+curcumin+on+bone+tissue+in+the+diabetic+rat:+Repair+of+peri-implant+and+critical-sized+defects.">Effect of curcumin on bone tissue in the diabetic rat: Repair of peri-implant and critical-sized defects.</a> Int J Oral Maxillofac Surg. 47 (11), 1495-1503 (2018).</li><li>De Oliveira, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+zoledronic+acid+and+dexamethasone+on+osseointegration+of+endosseous+implants:+Histological+and+histomorphometrical+evaluation+in+rats.">The effects of zoledronic acid and dexamethasone on osseointegration of endosseous implants: Histological and histomorphometrical evaluation in rats.</a> Clin Oral Implants Res. 26 (4), e17-e21 (2015).</li><li>Vandeweghe, S., Coelho, P. G., Vanhove, C., Wennerberg, A., Jimbo, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utilizing+micro-computed+tomography+to+evaluate+bone+structure+surrounding+dental+implants:+A+comparison+with+histomorphometry.">Utilizing micro-computed tomography to evaluate bone structure surrounding dental implants: A comparison with histomorphometry.</a> J Biomed Mater Res B Appl Biomater. 101 (7), 1259-1266 (2013).</li><li>Kleinert, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+obesity+and+diabetes+mellitus.">Animal models of obesity and diabetes mellitus.</a> Nat Rev Endocrinol. 14 (3), 140-162 (2018).</li><li>Grimm, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advancing+the+3Rs:+Innovation,+implementation,+ethics+and+society.">Advancing the 3Rs: Innovation, implementation, ethics and society.</a> Front Vet Sci. 10, 1185706 (2023).</li><li>Melcher, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+the+periosteum+in+repair+of+wounds+of+the+parietal+bone+of+the+rat.">Role of the periosteum in repair of wounds of the parietal bone of the rat.</a> Arch Oral Biol. 14 (9), 1101-1109 (1969).</li><li>Barrak, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heat+generation+during+guided+and+freehand+implant+site+preparation+at+drilling+speeds+of+1500+and+2000+rpm+at+different+temperatures:+An+in+vitro+study.">Heat generation during guided and freehand implant site preparation at drilling speeds of 1500 and 2000 rpm at different temperatures: An in vitro study.</a> Oral Health Prev Dent. 17 (4), 309-316 (2019).</li><li>Kniha, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+thermal+osteonecrosis+around+implants+in+the+rat+tibia:+Numerical+and+histomorphometric+results+in+context+of+implant+removal.">Effect of thermal osteonecrosis around implants in the rat tibia: Numerical and histomorphometric results in context of implant removal.</a> Sci Rep. 12 (1), 22227 (2022).</li><li>Da Silva, J. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apoptosis+in+bone+defect+of+diabetic+rats+treated+with+low+intensity+laser:+Radiological+and+immunohistochemical+approach.">Apoptosis in bone defect of diabetic rats treated with low intensity laser: Radiological and immunohistochemical approach.</a> International Journal of Morphology. 35, 178-183 (2017).</li><li>Zeller-Plumhoff, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+the+bone+ultrastructure+around+biodegradable+Mg-&#967;Gd+implants+using+small+angle+X-ray+scattering+and+X-ray+diffraction.">Analysis of the bone ultrastructure around biodegradable Mg-&#967;Gd implants using small angle X-ray scattering and X-ray diffraction.</a> Acta Biomater. 101, 637-645 (2020).</li><li>Bruns, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+material+dependency+of+peri-implant+morphology+and+stability+in+healing+bone.">On the material dependency of peri-implant morphology and stability in healing bone.</a> Bioact Mater. 28, 155-166 (2023).</li><li>De Morais, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+diabetes+mellitus+and+insulin+therapy+on+bone+density+around+osseointegrated+dental+implants:+A+digital+subtraction+radiography+study+in+rats.">Effect of diabetes mellitus and insulin therapy on bone density around osseointegrated dental implants: A digital subtraction radiography study in rats.</a> Clin Oral Implants Res. 20 (8), 796-801 (2009).</li><li>Aydemir Celep, N., Kara, H., Erbas, E., Dogan, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radioprotective+role+of+amifostine+on+osteointegration+of+titanium+implants+in+the+tibia+of+rats.">Radioprotective role of amifostine on osteointegration of titanium implants in the tibia of rats.</a> J Vet Sci. 24 (3), 35 (2023).</li><li>Delgado-Ruiz, R. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Slow+drilling+speeds+for+single-drill+implant+bed+preparation.+Experimental+in+vitro+study.">Slow drilling speeds for single-drill implant bed preparation. Experimental in vitro study.</a> Clin Oral Investig. 22 (1), 349-359 (2018).</li><li>Abdel Motagly, M., El Khadem, A., Abdel Rassoul, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+low-speed+drilling+without+irrigation,+versus+convencionaldrilling+with+irrigation+regarding+heat+generation+and+peri-implant+marginal+bone+loss+(randomised+clinical+trial).">Assessment of low-speed drilling without irrigation, versus convencionaldrilling with irrigation regarding heat generation and peri-implant marginal bone loss (randomised clinical trial).</a> Alex Dent J. 46 (2), 33-38 (2021).</li><li>Lelovas, P. P., Xanthos, T. T., Thoma, S. E., Lyritis, G. P., Dontas, I. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+laboratory+rat+as+an+animal+model+for+osteoporosis+research.">The laboratory rat as an animal model for osteoporosis research.</a> Comp Med. 58 (5), 424-430 (2008).</li></ol>

Although the rat is a widely used model for studying osseointegration, it is important to define and describe a reproducible surgical technique for adequately placing implants. Such a technique could serve as a guide for the scientific community. Moreover, the fact that certain diseases, such as osteoporosis and diabetes, alter bone metabolism implies stronger demands for correctly designing surgical procedures. The rat compares favorably with other animal models since it presents the main features of both osteoporosis (...

The common choice of rats as experimental subjects is due to the fact that they are easy to breed and relatively inexpensive compared to larger animal models. The emergence of new procedures, such as the reliable reproduction of a disorder, e.g., osteoporosis or diabetes, makes this model especially useful for analyzing the potential use of treatments and/or the influence of the disease in the biological response to drugs and surgical devices or procedures1,2.
The rat&#39;s bone mass gain occurs mostly during the first 6 months of life, although some researchers believe that the long bone grows constantly for at least a year with a progressive increase in length1. With aging, there is a transition from modeling to remodeling, which does not occur in all cases equally throughout the bones2. Female Sprague Dawley rats grow more slowly than male rats and achieve a lower peak in weight than male rats1. Continuous bone elongation and varied bone remodeling dynamics in rats are factors that have to be taken into account when addressing human health issues; however, it has not yet been possible to find any experimental research that shows either lifelong rat bone development or the species&#39; inability to remodel bone1. If the experimentation starts around 10 months of age, a margin of at least 1 mm from the growth plate of the tibia should be left intact due to this longitudinal bone growth, an issue to be considered in dental implant studies2. Hormones are also a key parameter in bone research since at 8 months of age, male rats were found to have 22% greater bone width and 33% greater breaking strength than females in the tibia3.
The reliable reproduction of a disorder is thus very important in orthopedics and implant dentistry since osseointegration of an orthopedic screw or a dental implant is a complex process that depends on numerous factors influencing the systemic response to the device implantation into the bone. Systemic disorders like osteoporosis and diabetes are known to affect the success rate in orthopedics and implant dentistry, so the reliable reproduction of those disorders in rat models can be applied to explore ways to overcome these limitations.
The rat tibia, due to the easy surgical access, moderate bone volume, and the flat shape on the medial plate, makes it suitable for surgical bone implantation experiments4,5, and it has been used in numerous research studies exploring the effects of implant surface on osseointegration4,5,6. A growing number of studies assess the effects on osseointegration of coatings and substances added to the implant surface in both healthy animals7 and in compromised animals affected by diabetes or osteoporosis8,9,10,11,12,13,14.
The number of implant devices placed in one rat&#39;s tibia is limited and can differ depending on the type of study. Depending on the number of implants or study conditions, the dimensions of the devices must be adapted to minimize surgical trauma. In studies with one implant, a nearly human-size implant can be placed (2.0 mm in diameter and 4 to 5 mm in length), and bi-cortical anchorage can be achieved6,7,15,16. The dimensions of the implants in multi-implant protocols should adopt an appropriate implant size (1.5 mm in diameter and 2.5 mm in length)4,17.
The present study aims to describe a standardized surgical protocol for titanium implant placement on the tibia of two rat models: the osteoporosis and the diabetes rat model. Moreover, this study permits testing the surgical protocol to assess different types of implant surface biofunctionalization and its effect on osseointegration.
A sample of 80 rats was divided into two groups. In group 1, 40 ovariectomized Sprague Dawley females and 5 sham animals were selected, with a mean weight of 484 g and a mean age of 12 weeks. Based on vendor recommendations (see Table of Materials), three months after neutering, the experiment started. This waiting period ensured the disappearance of sex hormones. Osteoporosis was confirmed at the time of surgery based on micro-computed tomography (micro-CT) bone analysis, which reflected an average of 20% bone loss compared to the sham group. Group 2 consisted of 40 BBDR (Bio Breeding Diabetes Resistant) genetically modified Sprague Dawley rats with type II diabetes. The mean weight was 730 g, and the average age was 12 weeks. Prior to surgery, the diabetic status was confirmed with three consecutive days of glucose measurements with results higher than 200 mg/dL. Glucose was measured with a glucometer in 6 h of fasting, and a blood drop was collected by tail puncture.
Grade 3 titanium implants measuring 2 mm in length and 1.8 mm in diameter were used. All implants were sterilized in clean room conditions, by being ultrasonically cleaned in cyclohexane (3 times for 2 min), acetone (once for 1 min), deionized water (3 times for 2 min), ethanol (3 times for 2 min), and acetone (3 times for 2 min) using an ultrasound bath (230 VAC, 50/60 Hz, 360 W). Then, the samples were dried with nitrogen gas, and a nitrogen beam at 0.5 bar was applied directly onto the samples. Prior to implantation, the implants were first soaked in deionized water and then immersed in 70% ethanol (v/v) for 10 min. After this, the implants were transferred to sterile microcentrifuge tubes, and kept under sterile conditions until the surgery.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>22 G needles+A2:C30</td>
			<td>Terumo</td>
			<td>NN-2238R</td>
			<td></td>
		</tr>
		<tr>
			<td>4/0 monofilament synthetic resorbable suture</td>
			<td>Braun ( MonoSyn)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL, 10 mL syringes</td>
			<td>Braun</td>
			<td>4617100V-02 4606051V</td>
			<td></td>
		</tr>
		<tr>
			<td>Adson forceps</td>
			<td>Ant&#227;o Medical</td>
			<td>Ref: A586</td>
			<td></td>
		</tr>
		<tr>
			<td>BBDR ( Biobreeding Diabetes Resistant ) Sprague Dawley Rats</td>
			<td>Janvier Labs</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Betadine</td>
			<td>Mylan</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Buprecare</td>
			<td>Animalcare (UK)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo Caliper 0-40 mm 15 cm angled</td>
			<td>UL AMIN Industries</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo Needle Holder</td>
			<td>Ant&#227;o Medical</td>
			<td>Ref: AM1702</td>
			<td></td>
		</tr>
		<tr>
			<td>Dental surgery scissors curved and straight</td>
			<td>Ant&#227;o Medical</td>
			<td>AMA603 / AMA600</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric shaver</td>
			<td>Oster Pro 3000i</td>
			<td>34264482227</td>
			<td></td>
		</tr>
		<tr>
			<td>Extra Fine Graefe Forceps</td>
			<td>F.S.T</td>
			<td>Ref: 11150-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze pads</td>
			<td>COVIDIEN</td>
			<td>441001</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucometer</td>
			<td>Menarini (Italy)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Helicoidal Drill / OSTEO-PIN DRILL &#216;1.6 mm</td>
			<td>soadco</td>
			<td>Ref. OS-8001</td>
			<td></td>
		</tr>
		<tr>
			<td>Implants / SCREW OSTEO-PIN &#216;1.8 x 2.0 mm</td>
			<td>soadco</td>
			<td>Ref. OS-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflo</td>
			<td>Le Vet Pharma (Netherlands)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Lance pilot drill / Lanceolate Drill (DS)</td>
			<td>soadco</td>
			<td>Ref. 10 02 01 T</td>
			<td></td>
		</tr>
		<tr>
			<td>Latex gloves - Surgical gloves sterile</td>
			<td>Hartmann</td>
			<td>Ref: 9426495</td>
			<td></td>
		</tr>
		<tr>
			<td>Lucas Surgical Curette</td>
			<td>Ant&#227;o Medical</td>
			<td>Ref: AMA940-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Metacam</td>
			<td>Boehringer Ingelheim(Germany)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro forceps straight</td>
			<td>nopa</td>
			<td>Ref: AB 542/12</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-CT scan( Quantum Fx microCT )</td>
			<td>Perkin Elmer (US)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Osteoporotic Sprague Dawley females Rats</td>
			<td>Janvier Labs</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Periosteal elevator -&#160; Molt 2-4</td>
			<td>Ant&#227;o Medical</td>
			<td>Ref: A1564</td>
			<td></td>
		</tr>
		<tr>
			<td>Physiologic solution for Irrigation</td>
			<td>Hygitech</td>
			<td>Ref:10238</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Blade Carbon Steel 15C</td>
			<td>Razor Med</td>
			<td>Ref: 02846</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Gauze Swabs</td>
			<td>Alledental</td>
			<td>Ref: 270712</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Irrigation system</td>
			<td>Hygitech</td>
			<td>Ref:HY1-110001D</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile towels (1 piece per animal)</td>
			<td>Dinarex</td>
			<td>4410</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical contra-angle handpiece</td>
			<td>W&#38;H</td>
			<td>Ref: WS-75 LED G</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical contra-angle handpiece</td>
			<td>W&#38;H</td>
			<td>SN 08877</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical contra-angle handpiece</td>
			<td>W&#38;H</td>
			<td>SN 01309</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical Electric Motor</td>
			<td>WH Implantmed Type: SI-1023</td>
			<td>&#160;Ref: 30288000</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scalpel handle</td>
			<td>AsaDental</td>
			<td>Ref: 0350-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Towel clamps</td>
			<td>Xelpov surgical</td>
			<td>AF-773-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic device</td>
			<td>J.P. Selecta, Abrera, Spain</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

surgical bone implantation technique for rat tibia models of diabetes and osteoporosis

The rat has long served as a valuable animal model in implant dentistry and orthopedics, particularly in studying the interactions between biomaterials and bone tissue. The rat's tibia is frequently chosen due to its easy surgical access through thin tissue layers (skin and muscle) and the flattened shape of its medial face, facilitating the surgical insertion of intraosseous devices. Additionally, this model enables the induction of specific diseases, mimicking various clinical conditions to assess biological responses to different implant conditions like geometry, surface texture, or biological cues. However, despite its robust cortical structure, certain intraosseous devices may require adaptations in design and size for successful implantation. Therefore, establishing standardized surgical methods for manipulating both soft and hard tissues in the implantation region is essential for ensuring proper implant or screw device placement, particularly in fields like implant dentistry and orthopedics. This study included eighty Sprague Dawley rats divided into two groups based on their respective diseases: Group 1 with osteoporosis and Group 2 with Type 2 Diabetes. Implantations were performed at 4 weeks and 12 weeks, with the same surgeon following a consistent surgical technique. A positive biological response was observed, indicating complete osseointegration of all implants placed. These results validate the success of the surgical protocol, which can be replicated for other studies and serve as a benchmark for the biomaterials community. Notably, osseointegration values remained stable at both 4 weeks and 12 weeks for both disease models, demonstrating a durable integration of the implant over time and emphasizing the establishment of an intimate bone connection as early as 4 weeks.

Surgical phase 
It is important to mention that both animal models used in this study present certain constraints due to the induced diseases. These constraints regarding the manipulation of hard and soft tissues are reflected during the surgical procedure.
In the diabetic model, the rat is larger, making it difficult to stabilize the legs during surgical procedures. This increases the surgical time and, consequently, the anesthesia time, which requires a longer recovery ...

The placement of implants in a rat model is an essential experimental procedure for clinical research. This study presents a comprehensive surgical protocol for implanting titanium implants into the tibia of rat models with diabetes and osteoporosis.

Surgical Bone Implantation Technique for Rat Tibia Models of Diabetes and Osteoporosis

The rat has long served as a valuable animal model in implant dentistry and orthopedics, particularly in studying the interactions between biomaterials ...

To begin the surgery on the anesthetized animal, make a one-centimeter long, full-thickness skin incision vertically along the proximal border of the anteromedial face of the tibia in the metaphysis region to expose the bone. Using small periosteal elevators, fully detach the tissue from the bone. Expose the bone and identify the insertion of the tibialis cranialis muscle, the gracilis, and the gastrocnemius lateral head muscle in the posterior border of the medial aspect of the tibia resembling a fibrous white tissue firmly adhered to the bone.Using a lance pilot drill at a depth of 2.4 millimeters with a maximum speed of 150 rotations per minute, drill the identified area as described previously under saline solution irrigation while avoiding any muscle injury. Perform a second drilling using a 1.6 millimeters diameter twist design drill at a depth of 2.4 millimeters. To place the titanium implant, first insert it with an intermediary piece attached to the 20:1 reduction contra angle.Next, clean the implant from any residual chemical sterilant by rotating it in the contra angle with simultaneous saline irrigation for 10 seconds. Then place the implant while making sure to stop the insertion before it completely passes the cortical bone where it is inserted. That is, the flat medial face of the tibia contiguous to the muscle's insertion.Suture the muscle tissue borders with simple internal sutures using a 4/0 monofilament synthetic resorbable glyconate suture. Finally, perform skin closure with an intradermal suture using the same 4/0 monofilament synthetic resorbable glyconate suture.

Research

JoVE Journal

Medicine

Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a ...

Models of Bone Metastasis

Bone metastases are a common occurrence in several malignancies, including breast, prostate, and lung. Once established in bone, tumors are responsible ...

Adjustable Stiffness, External Fixator for the Rat Femur Osteotomy and Segmental Bone Defect Models

The mechanical environment around the healing of broken bone is very important as it determines the way the fracture will heal. Over the past decade there ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Surgical Injury to the Mouse Pancreas through Ligation of the Pancreatic Duct as a Model for Endocrine and Exocrine Reprogramming and Proliferation

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide ...

Surgical Models of Gastroesophageal Reflux with Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

Surgical Training for the Implantation of Neocortical Microelectrode Arrays Using a Formaldehyde-fixed Human Cadaver Model

This protocol describes a procedure to assist surgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain. ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of ...

Vacuum-Sealed Hot Water Bath Immersion for the Preparation of Anatomical and Surgical Cadaveric Bone Models

Bone models serve many purposes, including improving anatomical understanding, preoperative surgical planning, and intraoperative referencing. Several ...