Ex Vivo and In Vivo Analysis of a Novel Porcine Aortic Patch for Vascular Reconstruction.

(1) Background: The aim of the present study was the biocompatibility analysis of a novel xenogeneic vascular graft material (PAP) based on native collagen won from porcine aorta using the subcutaneous implantation model up to 120 days post implantationem. As a control, an already commercially available collagen-based vessel graft (XenoSure®) based on bovine pericardium was used. Another focus was to analyze the (ultra-) structure and the purification effort. (2) Methods: Established methodologies such as the histological material analysis and the conduct of the subcutaneous implantation model in Wistar rats were applied. Moreover, established methods combining histological, immunohistochemical, and histomorphometrical procedures were applied to analyze the tissue reactions to the vessel graft materials, including the induction of pro- and anti-inflammatory macrophages to test the immune response. (3) Results: The results showed that the PAP implants induced a special cellular infiltration and host tissue integration based on its three different parts based on the different layers of the donor tissue. Thereby, these material parts induced a vascularization pattern that branches to all parts of the graft and altogether a balanced immune tissue reaction in contrast to the control material. (4) Conclusions: PAP implants seemed to be advantageous in many aspects: (i) cellular infiltration and host tissue integration, (ii) vascularization pattern that branches to all parts of the graft, and (iii) balanced immune tissue reaction that can result in less scar tissue and enhanced integrative healing patterns. Moreover, the unique trans-implant vascularization can provide unprecedented anti-infection properties that can avoid material-related bacterial infections.

[A novel tissue-engineered bone constructed by using human adipose-derived stem cells and biomimetic calcium phosphate scaffold coprecipitated with bone morphogenetic protein-2].

OBJECTIVE: To construct a novel biomimetic calcium phosphate (BioCaP) scaffold loaded with bone morphogenetic protein-2 (BMP-2), and to investigate its role in the osteogenesis of human adipose-derived stem cells (hASCs) in vitro and in vivo. METHODS: The BioCaP scaffold coprecipitated with BMP-2 (BMP-2-BioCaP) was constructed in this study. Field emission scanning electron microscopy (SEM) was used to analyze the morphology of the surfaces. The release kinetics was measured to evaluate the slow-release characteristics in vitro. BMP-2-BioCaP was immersed in proliferation medium (PM) or osteogenic medium (OM), respectively. The supernatants were collected and used to culture hASCs in vitro. Cell numbers were determined using the cell-counting kit-8 (CCK-8) to assess the cell proliferation. After 7 and 14 days, alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP. After 14 and 21 days, the calcification deposition was determined by alizarin red S (ARS) staining and quantification. The expressions of the osteoblast-related genes were tested on day 4 and day 14. In the in vivo study, 6 nude mice were used and implanted subcutaneously into the back of the nude mice for 4 groups: (1) BioCaP scaffold only, (2) BioCaP scaffold+hASCs, (3) BMP-2-BioCaP scaffold, (4) BMP-2-BioCaP scaffold+hASCs (test group). After 4 weeks of implantation, hematoxylin-eosin (HE) staining was performed to evaluate the in vivo osteogenesis of hASCs. RESULTS: SEM observations showed that BioCaP and BMP-2-BioCaP scaffold were entirely composed of straight, plate-like and sharp-edged crystal units, and the length of the crystal units varied between 5 and 10 µm. Release kinetics analysis demonstrated that BMP-2 incorporated with BioCaP could be released at certain concentration and last for more than 21 days, and the accumulative protein release could reach 20%. CCK-8 assays showed that cell proliferation was not significantly affected by BMP-2-BioCaP. ALP activity was higher by the induction of OM+BMP-2-BioCaP than of the other groups (P<0.01). More mineralization deposition and more expressions of osteoblast-related genes such as Runt-related transcription factor 2 (RUNX2), ALP, osteopontin (OPN) and osteocalcin (OC) were determined in the OM+BMP-2-BioCaP group at different time points (P<0.01). HE staining showed that, in the test group and BMP-2-BioCaP scaffold group, the extracellular matrix (ECM) with eosinophilic staining were observed around hASCs, and newly-formed bone-like tissues could be found in ECM around the scaffold materials. Moreover, compared with the BMP-2-BioCaP scaffold group, more bone-like tissues could be observed in ECM with typical structure of bone tissue in the test groups. No obvious positive results were found in the other groups. CONCLUSION: BMP-2-BioCaP scaffold could achieve slow-release of BMP-2 and promote the osteogenic differentiation of hASCs in vitro and in vivo. The novel tissue-engineered bone composed of hASCs and BMP-2-BioCaPis promising for the repair of bone defect.

Ridge preservation of compromised extraction sockets applying a soft cortical membrane: A canine proof-of-principle evaluation.

OBJECTIVES: To explore whether placement of a soft cortical membrane can restore and regenerate the original alveolar ridge contour in deficient sockets. MATERIALS AND METHODS: One Beagle dog was used in this proof-of-principle evaluation. In a first intervention, a standardized buccal dehiscence defect was artificially created at the distal roots of the 3rd and 4th mandibular premolars. Four weeks later, following endodontic treatment of the mesial roots, teeth were hemisected and the distal roots were extracted without raising a flap. A cortical membrane (Lamina®, Osteobiol) was placed outside of the bony envelope of the extraction socket to rebuild the buccal bone contour. Afterwards, sockets were filled with a collagen-modified porcine bone graft material (Gen-Os®, Osteobiol) to the level of the surrounding bone height. The socket orifice was closed with a porcine dermal matrix (Derma®). After four months, block specimens containing the socket-sites and remaining roots were retrieved, histologically processed and analyzed. RESULTS: Surgery and post-operative healing were uneventful. Histologically, bone formation under the membrane was found, i.e. bony protrusions and ossicles by osteoblasts could be identified. Concomitantly, the membrane showed clear signs of degradation. Bone substitute was well integrated in newly formed bone and resorption of particles was found. CONCLUSION: Three major observations were made in the present proof-of-principle study: (i) regeneration of a compromised socket seems possible when applying the presented approach, (ii) the soft cortical membrane was sufficiently stable to allow for the establishment of the contour and to inhibit soft tissue invasion and (iii) the applied xenogenic graft material was undergoing remodelling processes while allowing adequate bone regeneration.

Action of a deproteinized xenogenic biomaterial in the process of bone repair in rats submitted to inhalation of cigarette smoke.

PURPOSE: To investigate if the inorganic bovine bone matrix changes the bone formation in rats submitted to inhalation of cigarette smoke. METHODS: Twenty Wistar rats were divided into two groups: Cigarette Clot Group (CCG), which in the inhalation chamber received the smoke of 10 cigarettes, 3 times a day, 10 minutes, for 30 days and had the surgical cavity filled by clot; Cigarette Biomaterial Group (CBG), submitted to the same inhalation technique but with the cavity filled by biomaterial. RESULTS: In CCG there was a significant difference of new bone tissue in the analyzed periods (15 and 45 days), and in 15 days, there was 4.8 ± 0.42 of bone formed and 11.73 ± 0.59 (p <0.05) in 45 days. The CBG also showed a significant difference between the periods of 15 to 45 days, being respectively 6.16 ± 0.30 and 11.60 ± 0.61. However, when the groups were compared, within the same analyzed periods, a significant difference was observed only in the period of 15 days, with the new bone percentage being greater in the CBG. CONCLUSION: The bone matrix acted as an osteoinductive biomaterial, biocompatible and aided in the repair process, mainly in the initial period of recovery.

Biocompatibility and osteoconductivity of PLCL coated and noncoated xenografts: An in vitro and preclinical trial.

BACKGROUND: Cells, scaffolds, and growth factors are the key components in bone tissue engineering. Scaffold composition, topography, and architecture influence the amount of regenerated bone in the implantation site. The aims of the study were to compare viability and proliferation of mesenchymal stem cells (MSCs) seeded onto two commercial xenografts: Bio-Oss (BO) and bioactive bone bovine (BB). Next, these materials were compared for histomorphometric bone formation in a socket preservation model in rats. MATERIALS AND METHODS: MSCs were seeded onto monolayers of BO or BB granules. Cell viability and proliferation were evaluated after incubation of 0, 2, 20, and 48 h. A total of 24 Sprague Dawley rats underwent unilateral extraction of maxillary molars. Rats were randomly divided into three groups: natural healing (nongrafted socket) or socket preservation with either BO or BB. Rats were sacrificed after 8 weeks, and histomorphometric analysis was done to evaluate bone formation and residual scaffold at the extraction site. RESULTS: Differences in the metabolic activity of MSCs that were seeded onto BO or BB was observed at 2 h after seeding: the metabolic activity was elevated compared to baseline in the BB (P = .046) and not changed in the BO wells (P = .84). After 20 h, the metabolic activity of MSCs seeded onto BO was decreasing (P = .005), while cell viability was not changed in the BB group (P = .356). Intergroup comparison revealed higher metabolic activity of MSCs seeded on BB after 48 h compared with BO (P = .016). The in vivo results demonstrated differences in socket healing between the groups: percentage of new bone was higher in the BB compared to BO group (39.1 ± 14.3 vs. 23.7 ± 10.8%, respectively, P = .096). Connective tissue portion was higher in the BO group compared with BB (73.7 ± 11.1 vs. 49.6 ± 13.7%, respectively, P = .018). Residual grafting martial was higher in the BB (11.34 ± 4.18 vs. 2.62 ± 1.23%, P = .011). CONCLUSIONS: The results of this study demonstrating higher vitality and proliferation of MSCs seeded onto BB. Furthermore, following ridge preservation, higher percentage of new bone and lower residual scaffold were found in the BB compared with BO. This enhanced regenerative response might be the result of an enhancement of metabolic activity in cells attached to it. Further research will be needed to understand the precise mechanism.

Action of a deproteinized xenogenic biomaterial in the process of bone repair in rats submitted to inhalation of cigarette smoke

Abstract Purpose: To investigate if the inorganic bovine bone matrix changes the bone formation in rats submitted to inhalation of cigarette smoke. Methods: Twenty Wistar rats were divided into two groups: Cigarette Clot Group (CCG), which in the inhalation chamber received the smoke of 10 cigarettes, 3 times a day, 10 minutes, for 30 days and had the surgical cavity filled by clot; Cigarette Biomaterial Group (CBG), submitted to the same inhalation technique but with the cavity filled by biomaterial. Results: In CCG there was a significant difference of new bone tissue in the analyzed periods (15 and 45 days), and in 15 days, there was 4.8 ± 0.42 of bone formed and 11.73 ± 0.59 (p <0.05) in 45 days. The CBG also showed a significant difference between the periods of 15 to 45 days, being respectively 6.16 ± 0.30 and 11.60 ± 0.61. However, when the groups were compared, within the same analyzed periods, a significant difference was observed only in the period of 15 days, with the new bone percentage being greater in the CBG. Conclusion: The bone matrix acted as an osteoinductive biomaterial, biocompatible and aided in the repair process, mainly in the initial period of recovery.

Avaliação da remodelação do rebordo alveolar após exodontia minimamente traumática e utilização de enxerto xenógeno para preservação alveolar / Dimensional changes of alveolar bone in extraction sockets, after minimally traumatic extraction and socket fill with xenogenic bone

Introdução: Após a exodontia, a cicatrização natural do alvéolo provoca alterações dimensionais no rebordo alveolar, com diminuição dos tecidos moles e duros. Diferentes técnicas podem ser utilizadas para tentar diminuir essas alterações, com a intenção de facilitar a reabilitação estética e funcional com implantes, principalmente em áreas de dentes anteriores. Objetivos: Os objetivos deste estudo foram verificar a eficácia da utilização do material de origem xenógena Orthogen® na preservação do rebordo alveolar pós-exodontia e avaliar as características histológicas após 4 meses de reparo. Materiais e Métodos: 20 pacientes foram divididos aleatoriamente em 2 grupos, de acordo com a abordagem que iriam receber após a exodontia de uma raiz residual. No grupo teste, os alvéolos foram preenchidos com Orthogen® e selados com EGL (Enxerto Gengival Livre). No grupo controle, um coágulo foi preservado no alvéolo e o mesmo também foi selado com EGL, sem a utilização do Orthogen®. Em ambos os grupos as exodontias foram realizadas de maneira minimamente traumática, com o Kit de Extração Atraumática Neodent®. Modelos de gesso obtidos antes das exodontias (tempo 1) e após 4 meses (tempo 2), momentos antes da instalação dos implantes, foram utilizadas para as medidas dimensionais, através do escanemento dos modelos e medidas no programa Orthoanalyzer®. Amostras ósseas obtidas após 4 meses de reparo foram utilizadas para a análise histológica descritiva. Resultados: Os resultados demonstraram que, após 4 meses de reparo alveolar, os dois grupos apresentaram remodelação do rebordo alveolar e diminuição nas medidas realizadas. Porém, a remodelação do rebordo foi mais evidente no grupo controle (coágulo). Histologicamente foi possível observar formação de novo osso ao redor das partículas do Orthogen®. Conclusões: A utilização do Orthogen® para preenchimento dos alvéolos e preservação do rebordo alveolar foi eficaz na diminuição das...

Introduction: After tooth extraction, the natural healing of the socket is followed by dimensional changes of ridge contour, with marked reduce of hard and soft tissues. Socket reservation techniques can be used to improve functional and esthetics of the dental implants in this areas, especially in anterior teeth areas. Objectives: The aims of this study are to verify the efficacy of use Orthogen® to ridge preservation and analysis the histological characteristics after 4 months repair of the sockets. Material and Methods: 20 patients were randomly distributed in 2 groups, according with the treatments. In test group, the sockets were filled with Orthogen® and closed with a gingival free graft. In control group, the clots were preserved into the socket and free gingival graft was sutured in the deepithelialized marginal tissue of the socket. Stone casts were used to analyze the ridge dimensional changes with the Orthoanalyzer® software. After 4 months, bone samples were obtained during the dental implant procedure and were used to describe the histological characteristics. Results: The results showed that, after 4 months of wound healing, both groups showed a decrease in the measurements. However, the remodeling of the ridge was most evident in the control group (clot). Histological images showed the formation of new bone around the Orthogen® particles. Conclusion: The use of Orthogen® to socket preservation can collaborate to reduce the dimensional changes of the hard and soft tissue in post-extraction alveolar ridge sites.

Avaliação da remodelação do rebordo alveolar após exodontia minimamente traumática e utilização de enxerto xenógeno para preservação alveolar / Dimensional changes of alveolar bone in extraction sockets, after minimally traumatic extraction and socket fill with xenogenic bone

Introdução: Após a exodontia, a cicatrização natural do alvéolo provoca alterações dimensionais no rebordo alveolar, com diminuição dos tecidos moles e duros. Diferentes técnicas podem ser utilizadas para tentar diminuir essas alterações, com a intenção de facilitar a reabilitação estética e funcional com implantes, principalmente em áreas de dentes anteriores. Objetivos: Os objetivos deste estudo foram verificar a eficácia da utilização do material de origem xenógena Orthogen® na preservação do rebordo alveolar pós-exodontia e avaliar as características histológicas após 4 meses de reparo. Materiais e Métodos: 20 pacientes foram divididos aleatoriamente em 2 grupos, de acordo com a abordagem que iriam receber após a exodontia de uma raiz residual. No grupo teste, os alvéolos foram preenchidos com Orthogen® e selados com EGL (Enxerto Gengival Livre). No grupo controle, um coágulo foi preservado no alvéolo e o mesmo também foi selado com EGL, sem a utilização do Orthogen®. Em ambos os grupos as exodontias foram realizadas de maneira minimamente traumática, com o Kit de Extração Atraumática Neodent®. Modelos de gesso obtidos antes das exodontias (tempo 1) e após 4 meses (tempo 2), momentos antes da instalação dos implantes, foram utilizadas para as medidas dimensionais, através do escanemento dos modelos e medidas no programa Orthoanalyzer®. Amostras ósseas obtidas após 4 meses de reparo foram utilizadas para a análise histológica descritiva. Resultados: Os resultados demonstraram que, após 4 meses de reparo alveolar, os dois grupos apresentaram remodelação do rebordo alveolar e diminuição nas medidas realizadas. Porém, a remodelação do rebordo foi mais evidente no grupo controle (coágulo). Histologicamente foi possível observar formação de novo osso ao redor das partículas do Orthogen®. Conclusões: A utilização do Orthogen® para preenchimento dos alvéolos e preservação do rebordo alveolar foi eficaz na diminuição...

Introduction: After tooth extraction, the natural healing of the socket is followed by dimensional changes of ridge contour, with marked reduce of hard and soft tissues. Socket reservation techniques can be used to improve functional and esthetics of the dental implants in this areas, especially in anterior teeth areas. Objectives: The aims of this study are to verify the efficacy of use Orthogen® to ridge preservation and analysis the histological characteristics after 4 months repair of the sockets. Material and Methods: 20 patients were randomly distributed in 2 groups, according with the treatments. In test group, the sockets were filled with Orthogen® and closed with a gingival free graft. In control group, the clots were preserved into the socket and free gingival graft was sutured in the deepithelialized marginal tissue of the socket. Stone casts were used to analyze the ridge dimensional changes with the Orthoanalyzer® software. After 4 months, bone samples were obtained during the dental implant procedure and were used to describe the histological characteristics. Results: The results showed that, after 4 months of wound healing, both groups showed a decrease in the measurements. However, the remodeling of the ridge was most evident in the control group (clot). Histological images showed the formation of new bone around the Orthogen® particles. Conclusion: The use of Orthogen® to socket preservation can collaborate to reduce the dimensional changes of the hard and soft tissue in post-extraction alveolar ridge sites...