Vertical bone augmentation using collagenated or non-collagenated bone substitute materials with or without recombinant human bone morphogenetic protein-2 in a rabbit calvarial model.

PURPOSE: The aim of this study was to determine 1) the bone-regenerative effect of porcine bone block materials with or without collagen matrix incorporation, 2) the effect of a collagen barrier, and 3) the effect of adding recombinant human bone morphogenetic protein-2 (rhBMP-2) to the experimental groups. METHODS: Four treatment modalities were applied to rabbit calvaria: 1) deproteinized bovine bone mineral blocks (DBBM), 2) porcine bone blocks with collagen matrix incorporation (PBC), 3) porcine bone blocks alone without collagen matrix incorporation (PB), and 4) PBC blocks covered by a collagen membrane (PBC+M). The experiments were repeated with the addition of rhBMP-2. The animals were sacrificed after either 2 or 12 weeks of healing. Micro-computed tomography (micro-CT), histologic, and histomorphometric analyses were performed. RESULTS: Micro-CT indicated adequate volume stability in all block materials. Histologically, the addition of rhBMP-2 increased the amount of newly formed bone (NB) in all the blocks. At 2 weeks, minimal differences were noted among the NB of groups with or without rhBMP-2. At 12 weeks, the PBC+M group with rhBMP-2 presented the greatest NB (P<0.05 vs. the DBBM group with rhBMP-2), and the PBC and PB groups had greater NB than the DBBM group (P>0.05 without rhBMP-2, P<0.05 with rhBMP-2). CONCLUSIONS: The addition of rhBMP-2 enhanced NB formation in vertical augmentation using bone blocks, and a collagen barrier may augment the effect of rhBMP-2.

Immediate implant placement in conjunction with guided bone regeneration and/or connective tissue grafts: an experimental study in canines.

PURPOSE: This study was conducted to assess the effect of hard and/or soft tissue grafting on immediate implants in a preclinical model. METHODS: In 5 mongrel dogs, the distal roots of P2 and P3 were extracted from the maxilla (4 sites in each animal), and immediate implant placement was performed. Each site was randomly assigned to 1 of the following 4 groups: i) gap filling with guided bone regeneration (the GBR group), ii) subepithelial connective tissue grafting (the SCTG group), iii) GBR and SCTG (the GBR/SCTG group), and iv) no further treatment (control). Non-submerged healing was provided for 4 months. Histological and histomorphometric analyses were performed. RESULTS: Peri-implant tissue height and thickness favored the SCTG group (height of peri-implant mucosa: 1.14 mm; tissue thickness at the implant shoulder and ±1 mm from the shoulder: 1.14 mm, 0.78 mm, and 1.57 mm, respectively; median value) over the other groups. Bone grafting was not effective at the level of the implant shoulder and on the coronal level of the shoulder. In addition, simultaneous soft and hard tissue augmentation (the GBR/SCTG group) led to a less favorable tissue contour compared to GBR or SCTG alone (height of peri-implant mucosa: 3.06 mm; thickness of peri-implant mucosa at the implant shoulder and ±1 mm from the shoulder: 0.72 mm, 0.3 mm, and 1.09 mm, respectively). CONCLUSION: SCTG tended to have positive effects on the thickness and height of the peri-implant mucosa in immediate implant placement. However, simultaneous soft and hard tissue augmentation might not allow a satisfactory tissue contour in cases where the relationship between implant position and neighboring bone housing is unfavorable.

Effect of collagen membrane and of bone substitute on lateral bone augmentation with titanium mesh: An experimental in vivo study.

AIM: The aim of this study was to identify the additional effects of collagen membrane (CM) and of synthetic bone substitute (BS) on lateral bone augmentation of chronic peri-implant defect with titanium mesh (TM). MATERIALS AND METHODS: Atrophic alveolar ridge was induced in six canine mandibles, and 5 peri-implant defects were achieved in each hemi-mandible. Bone augmentation was attempted using the following randomly allocated modalities: (1) Control: no treatment, (2) TM only group: blood clot covered by TM, (3) TM+BS group: BS covered by TM, (4) TM+CM group: blood clot covered by TM and CM, and (5) TM+BS+CM group: BS covered by TM and CM. After 16 weeks of submerged healing, micro-CT and histomorphometric analyses were performed. RESULTS: TM exposure occurred in one case in the TM only group, one case in the TM+CM group, and two cases in the TM+BS+CM group. Histologically, pseudo-periosteum was observed along the inner and outer surfaces of TM, and the directions of the collagen fiber within the pseudo-periosteum differed according to the additional use of CM. In general, the TM only group rendered higher values in vertical defect fill and dimension of the augmented hard tissue in comparison with the other treatment groups. CONCLUSIONS: Due to the small sample size, this pilot study remains inconclusive. Within the limitations of the study, the use of CM and/or BS did not appear to have an additional benefit on lateral bone augmentation of peri-implant defect with TM.

Effectiveness of xenogeneic and synthetic bone-block substitute materials with/without recombinant human bone morphogenetic protein-2: A preclinical study using a rabbit calvarium model.

AIM: To investigate new bone (NB) formation by using bone-block substitute materials with/without recombinant human bone morphogenetic protein-2 (rhBMP-2). MATERIALS AND METHODS: Three synthetic bone-block substitute materials [biphasic calcium phosphate (BCP); nanostructured hydroxyapatite (NH); 3D-printed tricalcium phosphate/hydroxyapatite (3DP)] and one xenogeneic deproteinized bovine bone mineral (DBBM) block substitute were affixed to rabbit calvarium using osteosynthesis screws, either with rhBMP-2 (n = 12) or without rhBMP-2 (n = 16). At 2 or 12 weeks (n = 6 with rhBMP-2 and n = 8 without rhBMP-2 for each week), histologic, histomorphometric and microcomputed tomography analyses were performed. RESULTS: The application of rhBMP-2 increased NB formation in all experimental groups at both weeks. DBBM resulted in a greater area of NB compared with synthetic blocks either with or without rhBMP-2 at 2 weeks (2.8 ± 0.9 vs. 1.4 ± 0.5-1.9 ± 1.4 mm2 ; 1.4 ± 1.0 vs. 0.6 ± 0.3-0.9 ± 0.5 mm2 ) and without rhBMP-2 at 12 weeks (3.0 ± 0.8 vs. 1.7 ± 0.7-2.6 ± 1.5 mm2 ) (p > 0.05). NB formation did not differ significantly for DBBM and the three types of synthetic block with rhBMP-2 at 12 weeks (4.5 ± 2.0 vs. 3.8 ± 0.7-5.1 ± 1.1 mm2 ; p > 0.05). CONCLUSIONS: rhBMP-2 enhanced NB in all blocks. DBBM blocks yielded more NB than synthetic blocks without rhBMP-2. The application of rhBMP-2 appears to compensate for differences in late healing.

Programmed BMP-2 release from biphasic calcium phosphates for optimal bone regeneration.

This study aimed to fabricate a multi-layered biphasic calcium phosphate (BCP) platform for programmed bone morphogenetic protein-2 (BMP-2) release, which means to block the initial burst release and promote releasing during the differentiation phase of osteogenic cells. And it is to confirm in vivo whether this platform has osteogenic inductivity even when extremely low doses of BMP-2 are loaded compared to the conventional soaking method. Our strategy consisted of preparing a multilayer coating on BCP to minimize the contact between BMP-2 and BCP and allow the loading of BMP-2. The multilayer, which is surface-modified on BCP, is composed of an organosilicate and a natural polymer-based layer-by-layer (LbL) film. We applied (3-Aminopropyl)triethoxysilane (APTES) as an organosilicate was used for amine-functionalized BCP and (collagen/heparin)5 film was used to delay and sustain BMP-2 release. The coated multilayer not only reduced the initial burst release by more than 50% but also loaded more BMP-2. For in vivo experiment, histomorphometric analysis, it was observed that the BCP platform loaded with extremely low concentration BMP-2 (0.01 mg/ml) induced a significantly larger amount of new bones at 8 weeks compared to the conventional soaking method in the rabbit calvarium onlay graft model.

Immediate versus delayed application of bone morphogenetic protein-2 solution in damaged extraction sockets: a preclinical in vivo investigation.

OBJECTIVE: To compare the clinical, radiographic, and histological healing patterns between the immediate and delayed applications of bone morphogenetic protein-2 (BMP-2) in damaged extraction sockets in dogs. MATERIALS AND METHODS: The distal roots of the fourth premolars of the mandible were extracted bilaterally in five beagle dogs, and buccal bone defects (4 mm wide and 9 mm high) were surgically created. Collagenated biphasic calcium phosphate (CBCP) soaked for 10 min in 100 µL of BMP-2 solution was applied immediately to the defect site in the control group. In the test group, the BMP-2 solution of same dose was injected into the grafted site 2 weeks after grafting with a saline-soaked CBCP. The dogs were sacrificed 2 weeks later. Clinical, histological, and radiographic analyses were followed. RESULTS: Swelling and inflammatory reactions were predominantly observed in the control group at 2 weeks. The area of new bone formation was significantly larger in the control group compared with the test group (10.8 ± 7.0 mm2 [mean ± SD] and 6.3 ± 3.1 mm2, respectively; p = 0.043). No significant difference was found in ridge width at 2 mm, 4 mm and 6 mm below the lingual bone crest between the control (2.6 ± 1.0 mm, 3.2 ± 0.9 mm and 4.5 ± 0.5 mm, respectively) and test group (3.3 ± 1.0 mm, 3.7 ± 1.3 mm and 4.2 ± 1.0 mm; all p > 0.05). CONCLUSIONS: Delayed application of BMP-2 2 weeks after surgery did not show any advantage over immediate application of BMP-2 in terms of new bone formation. CLINICAL RELEVANCE: This study suggests that it might be better to apply BMP-2 immediately in alveolar ridge preservation, instead of delayed application, in order to enhance new bone formation.

Efficacy of Local Minocycline Agents in Treating Peri-Implantitis: An Experimental In Vivo Study in Beagle Dogs.

BACKGROUND: Local delivery agents (LDA) have the advantage of delivering the antibiotics at high concentrations to the targeted sites. However, the constant flow of gingival crevicular fluids and saliva may restrict their efficacy. Therefore, the drug sustainability and pharmacodynamic properties of any proposed LDA should be evaluated. METHODS: Four dental implants were placed unilaterally in the edentulous mandible of six beagle dogs. Peri-implantitis were experimentally induced using silk-ligatures. Each implant was randomly allocated to receive one of the following four treatments: (i) MC (Chitosan-alginate (CA) minocycline), (ii) MP (CA-without minocycline), (iii) PG (Polyacrylate-glycerin minocycline), and (iv) Control (mechanical debridement only). Mechanical therapies and LDAs were administered into the gingival sulcus two times at a 4-week interval. Drug sustainability as well as clinical, radiographical, and immunohistochemical (IHC) analyses were conducted to evaluate the efficacies of treatments. RESULTS: Reduced mean probing depth was observed in all of the test groups after the second delivery. A minimal marginal bone level change was observed during the treatment period (MP (-0.06 ± 0.53 mm) to PG (-0.25 ± 0.42 mm)). The distribution of IHC cell marker analysis of all targeted antibodies ranged from 6.34% to 11.33%. All treatment outcomes between the test groups were comparable. A prolonged retention of LDA was observed from CA microspheres (MC and MP) at both administrations (p < 0.017) and prolonged sustainability of bacteriostatic effect was observed from MC compared to PG after the second administration (p < 0.05). CONCLUSIONS: Prolonged retention of CA microspheres was observed and the longer bacteriostatic effect was observed from the MC group. Mechanical debridement with adjunct LDA therapy may impede peri-implantitis progression, however, prolonged drug action did not lead to improved treatment outcome.

Dimensional changes of the maxillary sinus augmented with a collagenated synthetic bone block or synthetic bone particulates: A pre-clinical study in rabbits.

OBJECTIVE: To compare the efficacy of a collagenated synthetic bone substitute (C-SBS) to a particulated synthetic bone substitute (P-SBS) in volume maintenance and new bone formations in a rabbit sinus model. MATERIALS AND METHODS: Either C-SBS or P-SBS was grafted in both sinuses of 16 rabbits. Four (N = 8) or 12 (N = 8) weeks after the surgery, total augmented volume (TAV) and area (TAA), as well as new bone volume (NBV) and area (NBA), were statistically compared by radiographic and histomorphometric analyses (p < .05). RESULTS: The differences in TAV, NBV, TAA and NBA between C-SBS and P-SBS groups at 4 weeks were not statistically significant. The TAV (267.13 ± 62.08 vs. 200.18 ± 40.32 mm3 ) and NBV (103.26 ± 10.50 vs. 71.10 ± 7.58 mm3 ) in group C-SBS were significantly higher than in group P-SBS at 12 weeks (p < .05). The TAA (19.36 ± 2.88 vs. 14.48 ± 2.08 mm2 ) and NBA (5.43 ± 1.20 vs. 3.76 ± 0.78 mm2 ) in group C-SBS were significantly higher than in group P-SBS at 12 weeks (p < .05). CONCLUSIONS: Collagenated synthetic bone substitute grafted in rabbit sinuses demonstrated more favourable outcomes across all outcome measures compared to P-SBS at 12 weeks.

Effects of soft tissue grafting prior to orthodontic treatment on preventing gingival recession in dogs.

PURPOSE: This study was conducted to assess the efficacy of prophylactic gingival grafting in the mandibular anterior labial area for preventing orthodontically induced gingival recession. METHODS: Eight mongrel dogs received gingival graft surgery at the first (I1) and third (I3) mandibular incisors on both sides based on the following group allocation: AT group (autogenous connective tissue graft on I1), AT-control group (contralateral side in the AT group), CM group (xenogeneic cross-linked collagen matrix graft on I3) and CM-control group (contralateral side in the CM group). At 4 weeks after surgery, 6 incisors were splinted and proclined for 4 weeks, followed by 16 weeks of retention. At 24 weeks after surgery, casts were made and compared with those made before surgery, and radiographic and histomorphometric analyses were performed. RESULTS: Despite the proclination of the incisal tip (by approximately 3 mm), labial gingival recession did not occur. The labial gingiva was thicker in the AT group (1.85±0.50 mm vs. 1.76±0.45 mm, P>0.05) and CM group (1.90±0.33 mm vs. 1.79±0.20 mm, P>0.05) than in their respective control groups. CONCLUSIONS: The level of the labial gingival margin did not change following labial proclination of incisors in dogs. Both the AT and CM groups showed enhanced gingival thickness.

Locally Applied Slow-Release of Minocycline Microspheres in the Treatment of Peri-Implant Mucositis: An Experimental In Vivo Study.

BACKGROUND: The objective of this is preclinical investigation was to evaluate the differential drug sustainability and pharmacodynamic properties of two local minocycline microsphere carriers: chitosan-coated alginate (CA) and poly(meth)acrylate-glycerin (PG). METHODS: Four dental implants were placed unilaterally in the edentulous mandible of six beagle dogs. Each implant was randomly assigned to receive one of the following four treatments: (i) CA (CA-based minocycline), (ii) placebo (CA substrate without minocycline), (iii) PG (PG-based minocycline) and (iv) control (mechanical debridement only). After inducing peri-implant mucositis, the randomly assigned treatments were administered into the gingival sulcus twice at a 4-week interval using a plastic-tipped syringe. Drug sustainability and pharmacodynamic (clinical, radiographical and cell marker intensity) evaluations were performed after each administration. RESULTS: The CA microspheres remained longer around the healing abutment compared to the PG microspheres at both administrations and a longer bacteriostatic effect was observed from CA (7.0 ± 5.7 days) compared to PG (1.2 ± 2.6 days). The efficacy of the applied therapies based on clinical, radiographical and histological analyses were comparable across all treatment groups. CONCLUSIONS: CA microspheres showed longer carrier and bacteriostatic effect sustainability when compared to PG microspheres, however, longer drug sustainability did not lead to improved treatment outcomes.

Volume stability of the augmented sinus using a collagenated bovine bone mineral grafted in case of a perforated Schneiderian membrane: An experimental study in rabbits.

OBJECTIVES: To determine the volume stability of a sinus augmented with a collagenated bovine bone mineral (CBBM) in case of an intact or perforated Schneiderian membrane (SM). MATERIALS AND METHODS: A bilateral sinus augmentation procedure was performed in eight rabbits. The SM was intentionally perforated in one side (SMP group), while it remained intact in contra-lateral side (control group) and the same amount of CBBM was then grafted. At 12 weeks, the animals were euthanized for radiographic and histomorphometric analyses. RESULTS: The augmented volume did not differ significantly between the two groups: 262.2 ± 32.1 mm3 in SMP group and 261.9 ± 48.5 mm3 in the control group (p = .959). There was no significant difference in the total augmented area: 24.7 ± 5.2 mm2 in SMP group and 23.2 ± 2.9 mm2 in the control group (p = .773). The areas of newly formed bone also did not differ significantly between the two groups, but was significantly lower at the centre of the augmented region than in the region of the surgical window in both groups (p < .05). CONCLUSION: A perforation of the SM in a rabbit model does neither impact the augmented volume nor new bone formation following grafting of the sinus with a CBBM.

Augmentation of keratinized tissue at tooth and implant sites by using autogenous grafts and collagen-based soft-tissue substitutes.

AIM: To investigate the effect of three treatment modalities on the gain of keratinized tissue (KT) at tooth and implant sites in dogs. MATERIALS AND METHODS: In five dogs, the distal roots of the mandibular second, third and fourth premolars were extracted, while the mesial roots were maintained. After 2 months of healing, implants were placed with KT excision. After another 2 months of healing, free gingival grafts, collagen-based matrices and apically positioned flap only were applied. The height of KT was measured during implant placement, immediately before soft-tissue grafting and after 10, 30 and 60 days. RESULTS: Two months after KT excision, spontaneous KT regrowth was greater at tooth sites than at implant sites (median, 2.0 mm vs. 1.1 mm). The outcomes of soft-tissue grafting at implant sites favoured the free gingival graft treatment, with a greater final median height (5.0-5.5 mm) and increase in KT (4.0-4.2 mm). Locations of the recipient sites significantly influenced KT regeneration at both tooth and implant sites. CONCLUSIONS: At implant sites, the free gingival graft treatment led to higher KT regeneration. At tooth sites, however, the differences between the three treatment modalities seemed clinically irrelevant.

The effects of hard and soft tissue grafting and individualization of healing abutments at immediate implants: an experimental study in dogs.

PURPOSE: To evaluate the effects of intra-alveolar socket grafting, subepithelial connective tissue grafts, and individualized abutments on peri-implant hard and soft tissue outcomes following immediate implant placement. METHODS: This randomized experimental study employed 5 mongrel dogs, with 4 sites per dog (total of 20 sites). The mesial roots of P3 and P4 were extracted in each hemimandible and immediate dental implants were placed. Each site was randomly assigned to 1 of 4 different treatment groups: standardized healing abutment (control group), alloplastic bone substitute material (BSS) + standardized healing abutment (SA group), BSS + individualized healing abutment (IA group), and BSS + individualized healing abutment + a subepithelial connective tissue graft (IAG group). Clinical, histological, and profilometric analyses were performed. The intergroup differences were calculated using the Bonferroni test, setting statistical significance at P<0.05. RESULTS: Clinically, the control and SA groups demonstrated a coronal shift in the buccal height of the mucosa (0.88±0.48 mm and 0.37±1.1 mm, respectively). The IA and IAG groups exhibited an apical shift of the mucosa (-0.7±1.15 mm and -1.1±0.96 mm, respectively). Histologically, the SA and control groups demonstrated marginal mucosa heights of 4.1±0.28 mm and 4.0±0.53 mm relative to the implant shoulder, respectively. The IA and IAG groups, in contrast, only showed a height of 2.6 mm. In addition, the height of the mucosa in relation to the most coronal buccal bone crest or bone substitute particles was not significantly different among the groups. Volumetrically, the IA group (-0.73±0.46 mm) lost less volume on the buccal side than the control (-0.93±0.44 mm), SA (-0.97±0.73 mm), and IAG (-0.88±0.45 mm) groups. CONCLUSIONS: The control group demonstrated the most favorable change of height of the margo mucosae and the largest dimensions of the peri-implant soft tissues. However, the addition of a bone substitute material and an individualized healing abutment resulted in slightly better preservation of the peri-implant soft tissue contour.

Tissue integration of zirconia and titanium implants with and without buccal dehiscence defects-A histologic and radiographic preclinical study.

OBJECTIVE: To histologically and radiographically evaluate soft (primary outcome) and hard tissue integration of two-piece titanium and zirconia dental implants with/without buccal dehiscence defects. MATERIALS & METHODS: In six dogs, five implants were randomly placed on both sides of the mandible: (a) Z1: a zirconia implant (modified surface) within the bony housing, (b) Z2: a zirconia implant (standard surface) within the bony housing, (c) T: a titanium implant within the bony housing, (d) Z1_D: a Z1 implant placed with a buccal bone dehiscence (3 mm in height, identical width to implant body), and (e) T_D: a titanium implant placed with a buccal bone dehiscence. Two weeks of healing and 6 months of loading were applied on each hemi-mandible, respectively. RESULTS: The median level of the margo mucosae shifted more apically over time in all groups (borderline statistical significance in groups Z1_D: -0.52 mm and T_D: -1.26 mm). The median height of the peri-implant mucosa in groups Z1_D and T_D was greatest at 2 weeks and 6 months, but the linear change in the peri-implant mucosa was statistically significant only for group T_D over time (-1.45 mm). Z1 demonstrated a higher bone-to-implant contact compared to Z2 and T. Minimal change of radiographic marginal bone levels in all groups was observed (<1 mm). CONCLUSION: When buccal dehiscence was presented, titanium implants presented significant loss of peri-implant mucosal height compared to zirconia implants with a modified surface, due to greater apical shift of the margo mucosae. A modified zirconia surface enhanced osseointegration.

Tissue integration of zirconia and titanium implants with and without buccal dehiscence defects.

PURPOSE: The purpose of the present study was to validate an experimental model for assessing tissue integration of titanium and zirconia implants with and without buccal dehiscence defects. METHODS: In 3 dogs, 5 implants were randomly placed on both sides of the mandibles: 1) Z1: a zirconia implant (modified surface) within the bony housing, 2) Z2: a zirconia implant (standard surface) within the bony housing, 3) T: a titanium implant within the bony housing, 4) Z1_D: a Z1 implant placed with a buccal bone dehiscence defect (3 mm), and 5) T_D: a titanium implant placed with a buccal bone dehiscence defect (3 mm). The healing times were 2 weeks (one side of the mandible) and 6 weeks (the opposite side). RESULTS: The dimensions of the peri-implant soft tissue varied depending on the implant and the healing time. The level of the mucosal margin was located more apically at 6 weeks than at 2 weeks in all groups, except group T. The presence of a buccal dehiscence defect did not result in a decrease in the overall soft tissue dimensions between 2 and 6 weeks (4.80±1.31 and 4.3 mm in group Z1_D, and 4.47±1.06 and 4.5±1.37 mm in group T_D, respectively). The bone-to-implant contact (BIC) values were highest in group Z1 at both time points (34.15%±21.23% at 2 weeks, 84.08%±1.33% at 6 weeks). The buccal dehiscence defects in groups Z1_D and T_D showed no further bone loss at 6 weeks compared to 2 weeks. CONCLUSIONS: The modified surface of Z1 demonstrated higher BIC values than the surface of Z2. There were minimal differences in the mucosal margin between 2 and 6 weeks in the presence of a dehiscence defect. The present model can serve as a useful tool for studying peri-implant dehiscence defects at the hard and soft tissue levels.