Porcine-derived Collagen Membrane Research Articles

Preclinical evaluation of mucogingival defect treatment using piscine membranes: An in vivo assessment of wound healing.

Periodontitis is a bacteria-induced chronic inflammatory disease characterized by degradation of the supporting tissue and bone in the oral cavity. Treatment modalities seek to facilitate periodontal rehabilitation while simultaneously preventing further gingival tissue recession and potentially bone atrophy. The aim of this study was to compare two differently sourced membranes, a resorbable piscine collagen membrane and a porcine-derived collagen membrane, in the repair of soft tissue defects utilizing a preclinical canine model. This in vivo component consisted of 10 beagles which were subjected to bilateral maxillary canine mucogingival flap defects, as well as bilateral soft tissue defects (or pouches) with no periodontal ligament damage in the mandibular canines. Defects received either a piscine-derived dermal membrane, (Kerecis® Oral, Ísafjörður, Iceland) or porcine-derived dermal membrane (Geistlich Mucograft®, Wolhusen, Switzerland) in a randomized fashion (to avoid site bias) and were allowed to heal for 30, 60, or 90 days. Statistical evaluation of tissue thickness was performed using general linear mixed model analysis of variance and least significant difference (LSD) post hoc analyses with fixed factors of time and membrane. Semi-quantitative analysis employed for inflammation assessment was evaluated using a chi-squared test along with a heteroscedastic t-test and values were reported as mean and corresponding 95% confidence intervals. In both the mucogingival flap defects and soft tissue gingival pouches, no appreciable qualitative differences were observed in tissue healing between the membranes. Furthermore, no statistical differences were observed in the thickness measurements between piscine- and porcine-derived membranes in the mucogingival flap defects (1.05 mm [±0.17] and 1.29 mm [±0.17], respectively [p = .06]) or soft tissue pouches (1.36 mm [±0.14] and 1.47 mm [±0.14], respectively [p = .27]), collapsed over time. Independent of membrane source (i.e., piscine or porcine), similar inflammatory responses were observed in both the maxilla and mandible at the three time points (p = .88 and p = .79, respectively). Histologic and histomorphometric evaluation results indicated that both membranes yielded equivalent tissue responses, remodeling dynamics and healing patterns for the mucogingival flap as well as the soft tissue gingival pouch defect models.

Osteogenic differentiation and reconstruction of mandible defects using a novel resorbable membrane: An in vitro and in vivo experimental study.

To evaluate the cellular response of both an intact fish skin membrane and a porcine-derived collagen membrane and investigate the bone healing response of these membranes using a translational, preclinical, guided-bone regeneration (GBR) canine model. Two different naturally sourced membranes were evaluated in this study: (i) an intact fish skin membrane (Kerecis Oral®, Kerecis) and (ii) a porcine derived collagen (Mucograft®, Geistlich) membrane, positive control. For the in vitro experiments, human osteoprogenitor (hOP) cells were used to assess the cellular viability and proliferation at 24, 48, 72, and 168 h. ALPL, COL1A1, BMP2, and RUNX2 expression levels were analyzed by real-time PCR at 7 and 14 days. The preclinical component was designed to mimic a GBR model in canines (n = 12). The first step was the extraction of premolars (P1-P4) and the 1st molars bilaterally, thereby creating four three-wall box type defects per mandible (two per side). Each defect site was filled with bone grafting material, which was then covered with one of the two membranes (Kerecis Oral® or Mucograft®). The groups were nested within the mandibles of each subject and membranes randomly allocated among the defects to minimize potential site bias. Samples were harvested at 30-, 60-, and 90-days and subjected to computerized microtomography (μCT) for three-dimensional reconstruction to quantify bone formation and graft degradation, in addition to histological processing to qualitatively analyze bone regeneration. Neither the intact fish skin membrane nor porcine-based collagen membrane presented cytotoxic effects. An increase in cell proliferation rate was observed for both membranes, with the Kerecis Oral® outperforming the Mucograft® at the 48- and 168-hour time points. Kerecis Oral® yielded higher ALPL expression relative to Mucograft® at both 7- and 14-day points. Additionally, higher COL1A1 expression was observed for the Kerecis Oral® membrane after 7 days but no differences were detected at 14 days. The membranes yielded similar BMP2 and RUNX2 expression at 7 and 14 days. Volumetric reconstructions and histologic micrographs indicated gradual bone ingrowth along with the presence of particulate bone grafts bridging the defect walls for both Kerecis Oral® and Mucograft® membranes, which allowed for the reestablishment of the mandible shape after 90 days. New bone formation significantly increased from 30 to 60 days, and from 60 to 90 days in vivo, without significant differences between membranes. The amount of bovine grafting material (%) within the defects significantly decreased from 30 to 90 days. Collagen membranes led to an upregulation of cellular proliferation and adhesion along with increased expression of genes associated with bone healing, particularly the intact fish skin membrane. Despite an increase in the bone formation rate in the defect over time, there was no significant difference between the membranes.

Fibrin glue does not promote migration and proliferation of bone marrow derived mesenchymal stem cells in collagenic membranes: an in vitro study

During Autologous Matrix-Induced Chondrogenesis (AMIC), the membrane is often glued into the chondral defect. However, whether fibrin glue influences cells proliferation and migration remain unclear. This study evaluated the impact of fibrin glue addition to biologic membranes loaded with bone marrow-derived mesenchymal stem cells (B-MSCs). A porcine derived collagen membrane (Cartimaix, Matricel GmbH, Germany) was used. B-MSCs were harvested from three different unrelated donors. The membranes were embedded in mounting medium with DAPI (ABCAM, Cambridge, UK) and analysed at 1-, 2-, 3-, 4-, 6-, and at 8-week follow-up. The DAPI ties the DNA of the cell nucleus, emitting blue fluorescence. DAPI/nuclei signals were analysed with fluorescence microscopy at 100-fold magnification. The group without fibrin glue demonstrated greater migration of the B-MSCs within the membrane at week 4 (P < 0.001), 6 (P < 0.001), and 8 (P < 0.001). No difference was found at week 1, 2, and 3. The group without fibrin glue demonstrated greater proliferation of B-MSCs within the membrane. These differences were significant at week 1 (P = 0.02), 2 (P = 0.008), 3 (P = 0.0009), 4 (P < 0.0001), 6 (P < 0.0001), 8 (P < 0.0001). Concluding, in the present setting, the use of fibrin in a collagenic biomembrane impairs B-MSCs proliferation and migration in vitro.

Proteomic Analysis of Porcine-Derived Collagen Membrane and Matrix.

Collagen membranes and matrices being widely used in guided bone regeneration and soft tissue augmentation have characteristic properties based on their composition. The respective proteomic signatures have not been identified. Here, we performed a high-resolution shotgun proteomic analysis on two porcine collagen-based biomaterials designed for guided bone regeneration and soft tissue augmentation. Three lots each of a porcine-derived collagen membrane and a matrix derived from peritoneum and/or skin were digested and separated by nano-reverse-phase high-performance liquid chromatography. The peptides were subjected to mass spectrometric detection and analysis. A total of 37 proteins identified by two peptides were present in all collagen membranes and matrices, with 11 and 16 proteins being exclusively present in the membrane and matrix, respectively. The common extracellular matrix proteins include fibrillar collagens (COL1A1, COL1A2, COL2A1, COL3A1, COL5A1, COL5A2, COL5A3, COL11A2), non-fibrillar collagens (COL4A2, COL6A1, COL6A2, COL6A3, COL7A1, COL16A1, COL22A1), and leucine-rich repeat proteoglycans (DCN, LUM, BGN, PRELP, OGN). The structural proteins vimentin, actin-based microfilaments (ACTB), annexins (ANXA1, ANXA5), tubulins (TUBA1B, TUBB), and histones (H2A, H2B, H4) were also identified. Examples of membrane-only proteins are COL12A1 and COL14A1, and, of matrix only proteins, elastin (ELN). The proteomic signature thus revealed the similarities between but also some individual proteins of collagen membrane and matrix.

Randomized clinical trial comparing PEG-based synthetic to porcine-derived collagen membrane in the preservation of alveolar bone following tooth extraction in anterior maxilla.

The objective of this randomized controlled trial was to compare alveolar ridge preservation using a bone substitute material and covered with a synthetic or porcine collagen membrane. Thirty-two sockets in the aesthetic maxillary region of 30 patients were randomized into two groups. Randomization was stratified according to bone wall defect. Flapless technique was used, and sockets were grafted with bi-phasic calcium phosphate particulate bone substitute and covered by synthetic polyethylene glycol (PEG; test group) or porcine-derived collagen membrane (CM; control group). No primary closure was attempted. A cone beam computed tomography (CBCT) scan was performed immediately after the surgical procedure and repeated 22weeks later. OnDemand3D was used to superimpose scan images and assess changes. The mean vertical and horizontal percentage bone loss were calculated and implants placed after 6months with or without additional augmentation. There were no baseline differences between groups or dropouts. The mean percentage loss at the labial plate and at the coronal part of the sockets was statistically significantly lower in the test group compared with controls (-2.86% [SD=13.48] versus 7.42% [SD=11.95]; 13.45% [SD=11.97] versus 28.59% [SD=16.97]). Implants were placed after 6months, and there was no difference in need for further augmentation between PEG (n=5) or CM (n=4). Sites treated with PEG membrane showed less percentage loss in horizontal and vertical measurements in this trial.

Antimicrobial Activity of an Amnion-Chorion Membrane to Oral Microbes.

Objective The aim of this study was to evaluate wound biomodification by assessing antimicrobial properties present within a human-derived composite amnion-chorion membrane (ACM). Methods Membranes analyzed were the human-derived ACM BioXclude™ and the porcine-derived collagen membrane Bio-Gide®. Paper discs with and without tetracycline served as positive and negative controls, respectively. The same number of colony-forming units per milliliter for each bacterial species (Aggregatibacter actinomycetemcomitans, Streptococcus mutans, and Streptococcus oralis) was inoculated on each of the discs. Discs from each group were removed at 12 and 24 hours and sonicated to remove the bacteria off the membranes. A serial dilution was performed to quantify bacterial growth. Results The ACM inhibited growth at all time points, with all bacterial strains, identical to the negative control tetracycline discs. The collagen membrane and positive controls did not inhibit growth of any of the bacterial species throughout the 24-hour study period. P < 0.05 for microbial growth on ACM or negative control vs. either collagen membrane or positive control. Conclusion ACM was proven to be as bactericidal as paper discs inoculated with tetracycline at its minimum bactericidal concentration. The ACM bactericidal property may be beneficial in the early wound healing process.

Effectiveness of Xenograft and Porcine-Derived Resorbable Membrane in Augmentation of Posterior Extraction Sockets with a Severe Wall Defect. A Radiographic/Tomographic Evaluation.

ABSTRACTObjectivesThe aim of the present prospective study was to evaluate, by means of intraoral periapical radiographs and cone-beam computed tomography, hard tissue changes after ridge augmentation procedures in posterior extraction sockets with severe wall defects. Material and MethodsTwenty patients, with a non-restorable premolar/molar tooth and severe wall defect, were enrolled in the present study, and underwent single-tooth extraction. Extraction sites were grafted with porcine-derived bone covered by porcine-derived collagen membrane. Intraoral periapical radiographs and cone-beam computed tomography scans, obtained at enrolment, and 6 months after ridge augmentation procedures were analysed and compared. ResultsIn the intraoral periapical radiographs, mean vertical bone gains detected at the distal, central and mesial aspects of the extraction sockets were 3.5 (SD 1.1) mm, 8.2 (SD 2.1) mm, and 3.9 (SD 1.7) mm, respectively. In the cone-beam computed tomography scans, the mean vertical bone gains detected at the more vestibular and more palatal aspects were 4.4 (SD 1.9) mm, and 3.3 (SD 2.8) mm, respectively. The mean horizontal bone gain was 3.5 (SD 1.6) mm. In all examined defects, mean vertical and horizontal bone levels showed a statistically significant increase (P < 0.05) at 6 months after extraction. ConclusionsWithin the limits of this study, the results suggest that porcine-derived bone graft covered by a collagen membrane can support significant vertical and horizontal bone gain at posterior post-extraction sockets with severe wall defects.

Contour changes after guided bone regeneration of large non-contained mandibular buccal bone defects using deproteinized bovine bone mineral and a porcine-derived collagen membrane: an experimental in vivo investigation.

The objective of this study was to evaluate soft tissue contour changes after three different regenerative therapies in chronic ridge defects. Buccal bone defects were created in the mandible of nine beagle dogs. Augmentation procedures were performed 3months later using a bone replacement graft (BRG), resorbable collagen membrane (MBG), or a combination of both procedures (CBG). Silicone impressions were taken before tooth extraction (T1), before the augmentation procedure (T2), and 3months after the regenerative surgeries (T3). Casts were optically scanned and stereolithography files were superimposed to analyze the horizontal changes in ridge contours. After defect creation, most part of the horizontal changes occurred 4 and 6mm below the gingival margin. In the mesial defect (D1) at T3, the mean horizontal gain in MBG amounted to 0.47±0.34mm, 0.79±0.67mm in the BRG, and 0.87±0.69mm for the CBG. In the middle defect (D2), the mean changes for the MBG were 0.11±0.31, 1.01±0.91 for the BRG, and 0.98±0.49 for the CBG. The mean changes in the distal defect (D3) amounted to 0.24±0.72 for the MBG, 1.04±0.92 for the BRG, and 0.86±0.56 for the CBG. The differences reached significance in all defects for the comparison MBG-BRG and MBG-CBG, while similar parameters were observed for the comparison BRG-CBG. BRG and CBG were equally effective and superior to MBG in increasing the horizontal tissue contours. The augmentation seldom reached the values before extraction. Scaffolding materials are needed for contour augmentation when using resorbable collagen membranes.

Extraction Socket Preservation Using Porcine-Derived Collagen Membrane Alone or Associated with Porcine-Derived Bone. Clinical Results of Randomized Controlled Study.

ABSTRACTObjectivesThe aim of present randomized controlled clinical trial was to clinically evaluate hard tissue changes after extraction socket preservation procedures compared to natural spontaneous healing.Material and MethodsThirty patients were enrolled in the present study and underwent single-tooth extraction in the premolar/molar areas. Ten sites were grafted with porcine-derived bone covered by collagen membrane, 10 covered by porcine-derived collagen membrane alone, and 10 underwent natural spontaneous healing. Vertical and horizontal bone changes after 3-month were evaluated at implant placement.ResultsThe vertical and horizontal bone changes at the extraction sockets treated with collagen membrane alone (vertical: -0.55 [SD 0.11] mm, and horizontal: -1.21 [SD 0.69] mm) and collagen membrane plus porcine-derived bone (vertical: -0.37 [SD 0.7] mm, and horizontal: -0.91 [SD 0.53] mm) were found significantly lower (P < 0.001), when compared to non-grafted sockets (vertical: -2.09 [SD 0.19] mm, and horizontal: -3.96 [SD 0.87] mm).In type 1 extraction sockets, in premolar sites, and in presence of vestibular bone thicknesses ≥ 1.5 mm, the use of collagen membrane alone revealed similar outcomes to those with additional graft material.ConclusionsAt the re-entry surgery, extraction sockets grafted with porcine-derived bone and covered by collagen membrane, and extraction sockets covered by porcine-derived collagen membrane alone, showed significantly lower vertical and horizontal bone changes, compared to extraction sockets sites underwent natural spontaneous healing. However, a complete prevention of remodelling is not achievable, irrespective of the technique used.

Effect of recombinant human bone morphogenic protein 9 (rhBMP9) loaded onto bone grafts versus barrier membranes on new bone formation in a rabbit calvarial defect model.

Recent research has demonstrated that recombinant human bone morphogenetic protein 9 (rhBMP9) has been considered the most osteoinductive growth factor of the BMP-family. In the present study, rhBMP9 was investigated for its influence in combination with two biomaterials for bone regenerative medicine. Either porcine-derived collagen membrane (CM) or deproteinized bovine bone mineral (DBM) combined with 20 µg of rhBMP9 were implanted in 6 mm rabbit calvarial defects. Bone augmentation was evaluated by microCT and histomorphometry at 8 weeks post-surgery. Both CM + rhBMP9 and DBM + rhBMP9 groups significantly promoted mineralized tissue volume (microCT) and area, new bone height and area (histomorphometric measurements) when compared to CM and DBM alone groups or control (empty). All specimens in the CM + rhBMP9 group but not all in the DBM + rhBMP9 group induced a complete horizontal bone defect closure. Multinucleated giant cells (MNGCs) were observed directly in contact with DBM surfaces irrespective of rhBMP9, whereas CM was generally not associated to the presence of MNGCs. When combined with rhBMP9, DBM augmented a larger volume of mineralized tissue (including the mineralized bone graft), whereas CM induced greater volume of native host bone. While DBM in combination with rhBMP9 induced higher mineralized tissue mostly associated with the bone grafting material, CM may have presented preferable results based on a higher horizontal defect closure with a faster regeneration of host new bone. The effect of including collagen within the carrier system of rhBMP9 on bone regeneration justifies further evaluation of this combination procedure in larger animal models. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2655-2661, 2017.

Maxillary sinus augmentation with autologous bone harvested from the lateral maxillary wall combined with bovine‐derived hydroxyapatite: clinical and histologic observations

The aim of this study was to determine the clinical efficacy of a mixture of autologous bone harvested from the lateral wall of the maxilla using bone shavers and bovine-derived hydroxyapatite (HA) placed as a graft to elevate the maxillary sinus floor. The histological picture of tissue found in the sinus, the survival rate and the success of the implants were all evaluated. A total of 90 titanium implants were placed in 34 patients. In all of them, the lateral maxillary wall was harvested as a particulate bone graft, subsequently mixed with bovine-derived HA and packed in the sinus cavity. The lateral access window was then covered with a bioresorbable porcine-derived collagen membrane. In 32 sinuses, a two-stage surgery was performed, while in the remaining 10 cases a one-stage surgery was carried out. In the two-stage approach, 14 randomly selected biopsies were obtained at the time of implant insertion after a healing period of 9 months. The histological specimens were histologically and histomorphometrically evaluated. One implant was lost, leading to a survival rate of 98.9%. The new bone consisted of lamellae of living bone contained osteocytes and in close contact with bovine bone particles that were partly infiltrated by newly formed bone. Bovine bone particle resorption could not be found. The histomorphometric analysis showed the following averages: 29% of newly formed bone and 21% of anorganic bovine bone. The marrow spaces made up the remaining 50% of the specimens. Sinus lift graft with autologous bone harvested from the maxillary lateral wall combined with demineralized bovine bone leads to a predictable outcome regarding the amount of bone formation in sinus floor augmentation.

Biological effects of a porcine-derived collagen membrane on intrabony defects

PurposeTo prolong the degradation time of collagen membranes, various cross-linking techniques have been developed. For cross-linking, chemicals such as formaldehyde and glutaraldehyde are added to collagen membranes, but these chemicals could adversely affect surrounding tissues. The aim of this study is to evaluate the ability of porous non-chemical cross-linking porcine-derived collagen nanofibrous membrane to enhance bone and associated tissue regeneration in one-wall intrabony defects in beagle dogs.MethodsThe second and third mandibular premolars and the first molars of 2 adult beagles were extracted bilaterally and the extraction sites were allowed to heal for 10 weeks. One-wall intrabony defects were prepared bilaterally on the mesial and distal side of the fourth mandibular premolars. Among eight defects, four defects were not covered with membrane as controls and the other four defects were covered with membrane as the experimental group. The animals were sacrificed 10 weeks after surgery.ResultsWound healing was generally uneventful. For all parameters evaluating bone regeneration, the experimental group showed significantly superior results compared to the control. In new bone height (NBh), the experimental group exhibited a greater mean value than the control (3.04 ± 0.23 mm/1.57 ± 0.59, P = 0.003). Also, in new bone area (NBa) and new bone volume (NBv), the experimental group showed superior results compared to the control (NBa, 34.48 ± 10.21% vs. 5.09 ± 5.76%, P = 0.014; and NBv, 28.04 ± 12.96 vs. 1.55 ± 0.57, P = 0.041). On the other hand, for parameters evaluating periodontal tissue regeneration, including junctional epithelium migration and new cementum height, there were no statistically significant differences between two groups.ConclusionsWithin the limitations of this study, this collagen membrane enhanced bone regeneration at one-wall intrabony defects. On the other hand, no influence of this membrane on periodontal tissue regeneration could be ascertained in this study.

The Use of Guided Tissue Regeneration Principles in Endodontic Surgery for Induced Chronic Periodontic‐Endodontic Lesions: A Clinical, Radiographic, and Histologic Evaluation

Chronic periodontic-endodontic lesions are not uncommon in clinical practice and their regenerative capacity has long been questioned. However, there are no published studies investigating the application of guided tissue regeneration techniques in combination with endodontic surgery using an induced perio-endo defect model. This study evaluated the clinical, radiographic, and histologic outcomes of three surgical procedures used to treat induced perio-endo lesions. Pulpal necrosis was induced in foxhounds along with surgical removal of radicular buccal bone. After 4 weeks, chronic lesions were clinically and radiographically assessed. Treatment surgery consisted of apicoectomy, root canal instrumentation, and retrofilling with mineral trioxide aggregate. Teeth were then assigned to one of the following treatment groups: open flap debridement only (OFD), OFD with bioabsorbable porcine-derived collagen membrane (BG), or OFD with BG and anorganic bovine bone matrix (BO/BG). Clinical parameters and standardized radiographs were assessed at defect creation; treatment surgery; and at 1, 2, 4, and 6 months. Animals were sacrificed at 6 months and specimens prepared for histometric analysis. Clinical and radiographic conditions improved during the study period. Mean epithelial attachment was similar between all groups. Mean connective tissue attachment for groups OFD, BG, and BO/BG was 3.79 mm, 2.63 mm, and 1.75 mm, respectively, and mean radicular bone height was 2.16 mm, 3.24 mm, and 3.45 mm, respectively. Statistically significant increases in the amount of new cementum were observed in groups BG and BO/BG when compared with OFD (P <0.05). Treatment of combined induced perio-endo lesions using bioabsorbable collagen membranes alone or in combination with anorganic bovine bone matrix resulted in increased amounts of bone, periodontal ligament, and significant increases in the amount of new cementum when compared to open flap debridement in a canine model.