Please activate JavaScript!
Please install Adobe Flash Player, click here for download
ePaper created 2015-11-17, 15:11:47 | version 1.38.0
44 Volume 1 | Issue 1/2015 Journal of Oral Science & Rehabilitation remaining bone height from measurement on a panoramic radiograph.37, 38 Exclusion criteria werethefollowing: antibiotic intake inthe previ- ous three months, prescription for more than six months of medications known to modify bone metabolism (e.g., bisphosphonates or corticos- teroids),pregnancyorintentiontobecomepreg- nantatthetimeofthescreening,thepresenceof anuntreatedchronicsinuscondition(e.g.,cystor tumor) or sepsis, a history of cancer or radiation to the oral cavity, complications of these condi- tions affecting the sinus area, and consumption of> 10cigarettes/day.Patientssmokingupto10 cigarettes/day39 andalcoholconsumerswerein- cluded in the study. For the statistical analysis, patients who smoked ≥ 1 cigarettes/day were considered smokers and those having ≥ 1 alco- hol-containingdrinks/day(> 10 gofalcohol/day)40 were considered alcohol consumers. Patients who met the inclusion and exclusion criteria were required to read, understand and sign the informed consent form before being enrolled in thestudy. S urgic a l proce du re s Patients were asked to take 875/125 mg amoxi- cillin/clavulanate (or, if allergic to penicillin, 300 mg clindamycin) t.i.d. for ten days, starting two days before the surgery to minimize infec- tionrisk.Allsurgicalprocedureswereperformed under local anesthesia (articaine with epineph- rine 40/0.01 mg/ml, Sanofi-Aventis Deutsch- land, Frankfurt/Main, Germany). The procedure proposed by Galindo-Moreno et al.41 was fol- lowed, using a bone scraper (Safescraper, Meta, Reggio Emilia, Italy)to harvestACBfromthe lat- eral wall and expose the Schneiderian mem- brane. After the membrane lift, sinus cavities were grafted with scraped ACB in combination with ABB particles sized between 250 and 1,000 μm (Geistlich Bio-Oss, Geistlich Pharma, Wolhusen, Switzerland);the ratio ofACBtoABB in the composite graft was 1:1 v/v.42 Amaximum of5 ccofgraftmaterialwasusedpersinuscavity. After bone grafting, an absorbable collagen membrane (Geistlich Bio-Gide, Geistlich Pharma, Wolhusen, Switzerland)was placed overthe lat- eral aspect ofthe bonywindow. Flaps were then carefully approximated and sutured with 3-0 surgical silk (Laboratorio Aragó, Barcelona, Spain)byprimaryintention. After a six-month healing period, a trephine (internal and external diameters of 3 mm and 4 mm, respectively) was used to harvest bone corebiopsiesfromthealveolarcrestinwhichim- plants were prosthetically planned. Implants (OsseoSpeed, Astra Tech, Mölndal, Sweden; Mi- crodent, Microdent Implant System, Barcelona, Spain)wereplacedinatwo-stageapproach. Hi sto lo gi cal stu dy Thetrephinebiopsieswerefixedin10%buffered formalin for 24h, decalcified in Decalcifier I (Surgipath Europe, Peterborough, UK), contain- ing formaldehyde (10% w/v), formic acid (8% w/v) and methanol (1% w/v), for 24 h at 37 ºC in an oven and embedded in paraffin. Then, 4 μm sections were cut along the central axis of the biopsies and dewaxed and hydrated for staining with hematoxylin–eosin, periodic acid–Schiff, Masson’s trichrome and Goldner’s trichrome. A millimeter scale in the eyepiece of a BH2 micro- scope (Olympus Optical, Tokyo, Japan) with a 40×objectivewasusedtocountosteoblasts,os- teoclasts and osteocytes per mm². Results were expressedintermsofthenumberofpositivecells permm2 . Bone histomorphometry was performed semi-automatically on Masson’s trichrome- stained sections, assessing ten randomized imageswith a 10× objective, using a microscope equipped with a digital camera (DP70, Olympus Optical, Tokyo, Japan) connected to a computer andapplyingImageJsoftware(Version1.48;de- velopedbytheU.S.NationalInstitutesofHealth, Bethesda, Md.). Separate quantifications ofvital bone, ABB particles and connective tissue were performed, expressing the results as percent- agesofeachcompartment. Immu no hi sto che mi cal analysi s Decalcified and paraffin-embedded sections were dewaxed, hydrated and heat-treated in 1 mMEDTAbufferforantigenicunmasking.Sec- tionswereincubatedfor60 minatroomtemper- ature with pre-diluted OPN polyclonal antibody toidentifycellularandinterstitialexpressionand with the following pre-diluted monoclonal anti- bodies (all from Master Diagnóstica, Granada, Spain): CD34 (clone QBEnd/10) to identify en- dothelial cells; CD56 (clone 56C04/123A8) to identify osteoblasts; tartrate-resistant acid phosphatase (TRAP; clone 26E5) to identify os- teoclasts; CD68 (clone KP1) to identify mono- cytesandmacrophages;andvimentin(cloneV9) Os teopon tin expres s io n i n ano rgani c bovi ne bo ne