An incision was made on the top of the alveolar crest with 1–2 releasing incisions at the adjacent teeth before the mucoperiosteal flap was elevated. The previously collected bone block graft was adjusted to the contour of the bone at the recipient site and fixated with 1–2 osteosynthesis screws (Walter Lorenz® Midface System, Biomet Microfixation, Jacksonville, USA) (Fig. 1 e). The remaining part of the autogenous bone graft was milled in a bone mill (Roswitha Quétin Dental-Produkte, Leimen, Germany), and autogenous bone graft particles were packed around the bone block. In the PRF group, three PRF membranes covered the grafted area (Fig. 1 f, g). In the control group, the grafted area was covered by deproteinised bovine bone mineral (Geistlich Bio-Oss® Spongiosa Granules, Geistlich Pharma AG, Wolhusen, Switzerland) and two layers of a resorbable native bilayer collagen membrane (Geistlich Bio-Gide®, Geistlich Pharma AG, Wolhusen, Switzerland). Finally, the periosteum of the mucoperiosteal flap was released by an incision to secure tension-free primary wound closure before suturing (4-0 Vicryl TM, Ethicon ®, Johnson & Johnson, NJ, USA) (Fig. 1 h).
Geistlich bio gide
Manufactured by Geistlich Pharma
Sourced in Switzerland
Geistlich Bio-Gide® is a collagen membrane designed for guided bone regeneration (GBR) and guided tissue regeneration (GTR) procedures. It serves as a scaffold to support and facilitate the regeneration of bone and soft tissue.
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13 protocols using geistlich bio gide
1
Alveolar Bone Grafting with PRF Membranes
2
Guided Bone Regeneration for Dental Implants
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Eight weeks after tooth extraction implants were placed. The implants placed had a diameter of 4.1 or 3.3 mm, and a length of 8 to 12 mm (Bone Level Tapered implants, Institute Straumann AG). After raising a mucoperiosteal flap from the top of the alveolar process, implants were placed according to the manufacturers guidelines. In case of thin a thin buccal plate (<1 mm) or a dehiscence at the buccal aspect, a guided bone regeneration (GBR) was performed. This implied coverage of the titanium surface of the dental implant with locally harvested autogenous bone, covered with DBBM (Geistlich Bio‐Oss®, Geistlich Pharma AG) and subsequent coverage with a resorbable membrane (Geistlich Bio‐Gide®, Geistlich Pharma AG). The autogenous bone chips were harvested via the existing flap or a relatively small extension of the flap.
3
Maxillofacial Bone Defect Reconstruction
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An experienced maxillo-facial surgeon (AnT) performed the osteotomies; a 20 mm long piece of the radius was removed mid-shaft with a Pietzo burr (Mectron, Carasco, Italy) under continuous saline cooling. A prefabricated template (4x6x20mm) was used as a guide. The removed bone was replaced with the same size CPC implant (n = 10, Fig 3 ) or AB (n = 10), after particulation into 1–3 mm bone chips using a manual bone grinder (The R. Quétin Bone-Mill, KLS Martin, Jacksonville, FL, USA). A collagenous membrane (25 x 25 mm, Geistlich Bio-Gide®, Geistlich Pharma AG, Wolhusen, Switzerland) was applied to form a cylinder around the AB in the defect area (Fig 4 ). No other fixation was used. The soft tissue was closed in separate layers using resorbable Monocryl 5–0 sutures (Ethicon, Johnsons & Johnson AB, Solna, Sweden). The surgery lasted 60–90 min.
4
Lateral Alveolar Bone Augmentation with CCXBB
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The surgical placement of the CCXBB blocks and the clinical evaluation has been described in detail in a previous publication [21 ]. In brief, severe alveolar horizontal bone deficiencies were isolated after rising full-thickness mucoperiosteal flaps. Once the horizontal width of the alveolar crest was measured 2 mm below the crest with a bone calliper bone blocks were shaped, pre-drilled and pre-hydrated for 5 min with sterile physiological saline before placement and were fixed with titanium osteosynthesis screws allowing for a stable contact between the block graft and the underlying bone. The spaces between the bone block and the surrounding bone were filled with DBBM particles (Geistlich Bio-Oss®, Geistlich Pharma AG, Wolhusen, Switzerland) and covered with a native collagen membrane (CM) (Geistlich Bio-Gide®, Geistlich Pharma AG, Wolhusen, Switzerland) fixed to the underlying bone with titanium tacks (FRIOS Fixation-Set®, SYMBIOS, Mainz, Germany). The muco-periosteal flaps were then coronally advanced and sutured achieving a tension-free primary closure (Fig. 2 ).![]()
Lateral bone augmentation of the alveolar crest (
5
Sericin-enhanced Bone Graft Materials
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Yeonnokjam was prepared by the Sericultural and Apicultural Division, National Institute of Agricultural Science, Rural Development Administration (RDA). The sericin from Yeonnokjam was treated by 4-hexylresorcinol (4HR) before degumming process to improve its protein conformation [16 (link)]. Gelatin sponge (Cutanplast Dental®, Uniplex, Sheffield, UK) with Yeonnokjam sericin (group G) and collagen membrane (Geistlich Bio-Gide®, Geistlich Pharma AG, Switzerland) with Yeonnokjam sericin (group C) were used as graft materials. All prepared grafts were fragmented for implanting into the bony defect. The detailed sample preparation procedure was shown in Fig. 1 . Each graft contained approximately 50 μg of sericin.![]()
Schematic drawings of experimental procedure
6
Bone Grafting and Implant Placement
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Depending on the quantity of bone, implants were placed following one of the following two procedures:
• GBR: With simultaneous GBR for the treatment of bone defects including dehiscences and infrabony defects. The GBR procedure involved grafting with a particulate deproteinized bovine bone mineral (DBBM) (Geistlich Bio-Oss ® spongiosa granules, Geistlich Pharma AG, Wolhusen, Switzerland), autogenous bone harvested from the site of surgery or a mixture of the two. The site was covered with a native bilayer collagen membrane (Geistlich Bio-Gide ® , Geistlich Pharma AG, Wolhusen, Switzerland).
• Control: Standard implant placement executed in situations with bone volume sufficient for complete coverage of the endosseous implant surface.
All implants exhibited a machined endosseous surface (Brånemark System, Nobel Biocare, Kloten, Switzerland). The placement of implants was performed either as type 2, type 3, or type 4 procedure (Hammerle et al. 2004 ). The implants were primarily covered for submerged healing and loaded after a minimum healing time of 6 months. There was one exception, where healing was obtained with the implant in the transmucosal position and prosthetic loading 3 weeks after implant placement. This implant was lost after 4 months and, therefore, not included in the clinical and radiographic analyses.
• GBR: With simultaneous GBR for the treatment of bone defects including dehiscences and infrabony defects. The GBR procedure involved grafting with a particulate deproteinized bovine bone mineral (DBBM) (Geistlich Bio-Oss ® spongiosa granules, Geistlich Pharma AG, Wolhusen, Switzerland), autogenous bone harvested from the site of surgery or a mixture of the two. The site was covered with a native bilayer collagen membrane (Geistlich Bio-Gide ® , Geistlich Pharma AG, Wolhusen, Switzerland).
• Control: Standard implant placement executed in situations with bone volume sufficient for complete coverage of the endosseous implant surface.
All implants exhibited a machined endosseous surface (Brånemark System, Nobel Biocare, Kloten, Switzerland). The placement of implants was performed either as type 2, type 3, or type 4 procedure (Hammerle et al. 2004 ). The implants were primarily covered for submerged healing and loaded after a minimum healing time of 6 months. There was one exception, where healing was obtained with the implant in the transmucosal position and prosthetic loading 3 weeks after implant placement. This implant was lost after 4 months and, therefore, not included in the clinical and radiographic analyses.
7
Guided Bone Regeneration for Dental Implants
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From 1994 to 1996, 72 patients received 265 implants. Of these, 256 were Brånemark implants (Nobel Biocare Services AG, Göteborg, Sweden), followed by 8 Biomed 3i (Biomet 3i, Palm Beach Gardens, FL, USA) and one single IMZ implant (Friatec® AG). Out of 265 implants, 112 implants were treated with a resorbable membrane (BG) and 41 with a nonresorbable (GT). The remaining 112 implants were placed in pristine bone and did not require further regenerative therapies. These are considered as the control group (CT).
Group BG: Sites with a dehiscence defect following implant placement, treated with GBR using deproteinized bovine bone mineral (Geistlich Bio‐Oss®, Geistlich Pharma AG, Wolhusen, Switzerland) and a resorbable, native collagen membrane (Geistlich Bio‐Gide®, Geistlich Pharma AG, Wolhusen, Switzerland).
Group GT: Sites with a dehiscence defect following implant placement, treated with GBR using deproteinized bovine bone mineral (Geistlich Bio‐Oss®, Geistlich Pharma AG) and a nonresorbable membrane made of expanded polytetrafluorethylene (e‐PTFE; Gore‐Tex®, W.L. Gore/Implant Innovations, West Palm Beach, FL, USA).
Group CT: Implants placed in pristine bone without regenerative therapy.
Further details on the original procedures and randomization processes were reported earlier (Zitzmann et al., 1997 (link), 2001 (link)).
Group BG: Sites with a dehiscence defect following implant placement, treated with GBR using deproteinized bovine bone mineral (Geistlich Bio‐Oss®, Geistlich Pharma AG, Wolhusen, Switzerland) and a resorbable, native collagen membrane (Geistlich Bio‐Gide®, Geistlich Pharma AG, Wolhusen, Switzerland).
Group GT: Sites with a dehiscence defect following implant placement, treated with GBR using deproteinized bovine bone mineral (Geistlich Bio‐Oss®, Geistlich Pharma AG) and a nonresorbable membrane made of expanded polytetrafluorethylene (e‐PTFE; Gore‐Tex®, W.L. Gore/Implant Innovations, West Palm Beach, FL, USA).
Group CT: Implants placed in pristine bone without regenerative therapy.
Further details on the original procedures and randomization processes were reported earlier (Zitzmann et al., 1997 (link), 2001 (link)).
8
Gingiva Tissue Culture: Ex Vivo Analysis
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Gingiva tissue culture was performed to represent an ex vivo situation.
Gingiva tissue from extracted teeth was cut into pieces of approximately 1 mm3 diameter to match the diameter of three GC spheroids. Tissue pieces were seeded onto 1 cm2 of a collagen membrane (Geistlich Bio‐Gide®; Geistlich Pharma AG) or bone substitute (Geistlich Bio‐Oss®; Geistlich Pharma AG) covering 1 cm2 of plastic. Gingiva ex vivo tissues were cultured for 24 h on each surface.
Tissue pieces of gingiva from four different donors were used for histological and immunohistochemical analysis (n = 4), and tissue pieces of gingiva from six different donors were used for metabolic activity, mRNA level and protein analysis (n = 6).
Gingiva tissue from extracted teeth was cut into pieces of approximately 1 mm3 diameter to match the diameter of three GC spheroids. Tissue pieces were seeded onto 1 cm2 of a collagen membrane (Geistlich Bio‐Gide®; Geistlich Pharma AG) or bone substitute (Geistlich Bio‐Oss®; Geistlich Pharma AG) covering 1 cm2 of plastic. Gingiva ex vivo tissues were cultured for 24 h on each surface.
Tissue pieces of gingiva from four different donors were used for histological and immunohistochemical analysis (n = 4), and tissue pieces of gingiva from six different donors were used for metabolic activity, mRNA level and protein analysis (n = 6).
9
Gingival Cell Culture on Biomaterials
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Gingiva monolayer culture was performed to represent the traditional in vitro 2D cell culture.
A cell suspension of 50,000 human GC in α‐minimal essential medium with FCS and antibiotics was seeded onto 1 cm2 of a collagen membrane (Geistlich Bio‐Gide®; Geistlich Pharma AG, Wolhusen, Switzerland) or bone substitute (Geistlich Bio‐Oss®; Geistlich Pharma AG) covering 1 cm2 of plastic. The same amount of GC was seeded onto the plastic surface of a cell culture plate as control. GC monolayers were cultured for 24 h on each surface.
GC from 6 different donors were used for histological and immunohistochemical analysis (n = 6), and GC from 6 different donors were used for metabolic activity, mRNA level and protein analysis (n = 6).
A cell suspension of 50,000 human GC in α‐minimal essential medium with FCS and antibiotics was seeded onto 1 cm2 of a collagen membrane (Geistlich Bio‐Gide®; Geistlich Pharma AG, Wolhusen, Switzerland) or bone substitute (Geistlich Bio‐Oss®; Geistlich Pharma AG) covering 1 cm2 of plastic. The same amount of GC was seeded onto the plastic surface of a cell culture plate as control. GC monolayers were cultured for 24 h on each surface.
GC from 6 different donors were used for histological and immunohistochemical analysis (n = 6), and GC from 6 different donors were used for metabolic activity, mRNA level and protein analysis (n = 6).
10
Gingival Spheroid Culture on Biomaterials
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Gingiva spheroid culture was performed to represent an in vivo‐like 3D cell culture.
GC spheroids were produced using 3D Petri Dishes® (Microtissues, Inc., Providence, RI, USA), following the instructions of the manufacturer (Janjić, Lilaj, Moritz, & Agis,2018 ). Three GC spheroids with a total cell number of 50,000 and an average diameter of 356 μm/spheroid were seeded onto 1 cm2 of a collagen membrane (Geistlich Bio‐Gide®; Geistlich Pharma AG) or bone substitute (Geistlich Bio‐Oss®; Geistlich Pharma AG) covering 1 cm2 of plastic. Three GC spheroids with the same amount of cells were seeded onto the plastic surface of a cell culture plate as control. The three GC spheroids together approximately equaled the total diameter of one piece of gingiva ex vivo tissue. GC spheroids were cultured for 24 h on each surface.
GC from 6 donors were used for histological and immunohistochemical analysis (n = 6) as well as metabolic activity, mRNA level and protein analysis (n = 6).
GC spheroids were produced using 3D Petri Dishes® (Microtissues, Inc., Providence, RI, USA), following the instructions of the manufacturer (Janjić, Lilaj, Moritz, & Agis,
GC from 6 donors were used for histological and immunohistochemical analysis (n = 6) as well as metabolic activity, mRNA level and protein analysis (n = 6).
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